Sample records for lateral flow chromatography

  1. Lateral flow strip assay

    DOEpatents

    Miles, Robin R [Danville, CA; Benett, William J [Livermore, CA; Coleman, Matthew A [Oakland, CA; Pearson, Francesca S [Livermore, CA; Nasarabadi, Shanavaz L [Livermore, CA

    2011-03-08

    A lateral flow strip assay apparatus comprising a housing; a lateral flow strip in the housing, the lateral flow strip having a receiving portion; a sample collection unit; and a reagent reservoir. Saliva and/or buccal cells are collected from an individual using the sample collection unit. The sample collection unit is immersed in the reagent reservoir. The tip of the lateral flow strip is immersed in the reservoir and the reagent/sample mixture wicks up into the lateral flow strip to perform the assay.

  2. A lateral electrophoretic flow diagnostic assay

    PubMed Central

    Lin, Robert; Skandarajah, Arunan; Gerver, Rachel E.; Neira, Hector D.; Fletcher, Daniel A.

    2015-01-01

    Immunochromatographic assays are a cornerstone tool in disease screening. To complement existing lateral flow assays (based on wicking flow) we introduce a lateral flow format that employs directed electrophoretic transport. The format is termed a “lateral e-flow assay” and is designed to support multiplexed detection using immobilized reaction volumes of capture antigen. To fabricate the lateral e-flow device, we employ mask-based UV photopatterning to selectively immobilize unmodified capture antigen along the microchannel in a barcode-like pattern. The channel-filling polyacrylamide hydrogel incorporates a photoactive moiety (benzophenone) to immobilize capture antigen to the hydrogel without a priori antigen modification. We report a heterogeneous sandwich assay using low-power electrophoresis to drive biospecimen through the capture antigen barcode. Fluorescence barcode readout is collected via a low-resource appropriate imaging system (CellScope). We characterize lateral e-flow assay performance and demonstrate a serum assay for antibodies to the hepatitis C virus (HCV). In a pilot study, the lateral e-flow assay positively identifies HCV+ human sera in 60 min. The lateral e-flow assay provides a flexible format for conducting multiplexed immunoassays relevant to confirmatory diagnosis in near-patient settings. PMID:25608872

  3. Field-flow fractionation and hydrodynamic chromatography on a microfluidic chip.

    PubMed

    Shendruk, Tyler N; Tahvildari, Radin; Catafard, Nicolas M; Andrzejewski, Lukasz; Gigault, Christian; Todd, Andrew; Gagne-Dumais, Laurent; Slater, Gary W; Godin, Michel

    2013-06-18

    We present gravitational field-flow fractionation and hydrodynamic chromatography of colloids eluting through 18 μm microchannels. Using video microscopy and mesoscopic simulations, we investigate the average retention ratio of colloids with both a large specific weight and neutral buoyancy. We consider the entire range of colloid sizes, including particles that barely fit in the microchannel and nanoscopic particles. Ideal theory predicts four operational modes, from hydrodynamic chromatography to Faxén-mode field-flow fractionation. We experimentally demonstrate, for the first time, the existence of the Faxén-mode field-flow fractionation and the transition from hydrodynamic chromatography to normal-mode field-flow fractionation. Furthermore, video microscopy and simulations show that the retention ratios are largely reduced above the steric-inversion point, causing the variation of the retention ratio in the steric- and Faxén-mode regimes to be suppressed due to increased drag. We demonstrate that theory can accurately predict retention ratios if hydrodynamic interactions with the microchannel walls (wall drag) are added to the ideal theory. Rather than limiting the applicability, these effects allow the microfluidic channel size to be tuned to ensure high selectivity. Our findings indicate that particle velocimetry methods must account for the wall-induced lag when determining flow rates in highly confining systems.

  4. Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cary, Robert E.

    2015-12-08

    Highly simplified lateral flow chromatographic nucleic acid sample preparation methods, devices, and integrated systems are provided for the efficient concentration of trace samples and the removal of nucleic acid amplification inhibitors. Methods for capturing and reducing inhibitors of nucleic acid amplification reactions, such as humic acid, using polyvinylpyrrolidone treated elements of the lateral flow device are also provided. Further provided are passive fluid control methods and systems for use in lateral flow assays.

  5. Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cary, Robert B.

    Highly simplified lateral flow chromatographic nucleic acid sample preparation methods, devices, and integrated systems are provided for the efficient concentration of trace samples and the removal of nucleic acid amplification inhibitors. Methods for capturing and reducing inhibitors of nucleic acid amplification reactions, such as humic acid, using polyvinylpyrrolidone treated elements of the lateral flow device are also provided. Further provided are passive fluid control methods and systems for use in lateral flow assays.

  6. Molecular characterization of branched polysaccharides from Tremella fuciformis by asymmetrical flow field-flow fractionation and size exclusion chromatography.

    PubMed

    Wu, Ding-Tao; Deng, Yong; Zhao, Jing; Li, Shao-Ping

    2017-11-01

    To accurately characterize branched polysaccharides with high molecular weights from medicinal and edible mushrooms and identify the limitations of size exclusion chromatography, molecular characteristics of polysaccharides from Tremella fuciformis were determined and compared by asymmetrical flow field-flow fractionation coupled with multiangle laser light scattering and refractive index detection, and size exclusion chromatography coupled with multiangle laser light scattering and refractive index detection, respectively. Results showed that molecular weights of three batches of T. fuciformis polysaccharides were determined as 2.167 × 10 6 (TF1), 2.334 × 10 6 (TF2), and 2.435 × 10 6  Da (TF3) by size exclusion chromatography, and 3.432 × 10 6 (TF1), 3.739 × 10 6 (TF2), and 3.742 × 10 6  Da (TF3) by asymmetrical flow field-flow fractionation, as well as 3.469 × 10 6  Da (TF1) by off-line multiangle laser light scattering, respectively. Results suggested that size exclusion chromatography was unable to accurately characterize T. fuciformis polysaccharides, which may be due to its limitations such as shear degradation and abnormal coelution. Compared to size exclusion chromatography, asymmetrical flow field-flow fractionation could be a better technique for the molecular characterization of branched polysaccharides with high molecular weights from medicinal and edible mushrooms, as well as from other natural resources. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Battery operated preconcentration-assisted lateral flow assay.

    PubMed

    Kim, Cheonjung; Yoo, Yong Kyoung; Han, Sung Il; Lee, Junwoo; Lee, Dohwan; Lee, Kyungjae; Hwang, Kyo Seon; Lee, Kyu Hyoung; Chung, Seok; Lee, Jeong Hoon

    2017-07-11

    Paper-based analytical devices (e.g. lateral flow assays) are highly advantageous as portable diagnostic systems owing to their low costs and ease of use. Because of their low sensitivity and detection limits for biomolecules, these devices have several limitations in applications for real-field diagnosis. Here, we demonstrate a paper-based preconcentration enhanced lateral flow assay using a commercial β-hCG-based test. Utilizing a simple 9 V battery operation with a low power consumption of approximately 81 μW, we acquire a 25-fold preconcentration factor, demonstrating a clear sensitivity enhancement in the colorimetric lateral flow assay; consequently, clear colors are observed in a rapid kit test line, which cannot be monitored without preconcentration. This device can also facilitate a semi-quantitative platform using the saturation value and/or color intensity in both paper-based colorimetric assays and smartphone-based diagnostics.

  8. Variable parameter McCarthy-Muskingum routing method considering lateral flow

    NASA Astrophysics Data System (ADS)

    Yadav, Basant; Perumal, Muthiah; Bardossy, Andras

    2015-04-01

    The fully mass conservative variable parameter McCarthy-Muskingum (VPMM) method recently proposed by Perumal and Price (2013) for routing floods in channels and rivers without considering lateral flow is extended herein for accounting uniformly distributed lateral flow contribution along the reach. The proposed procedure is applied for studying flood wave movement in a 24.2 km river stretch between Rottweil and Oberndorf gauging stations of Neckar River in Germany wherein significant lateral flow contribution by intermediate catchment rainfall prevails during flood wave movement. The geometrical elements of the cross-sectional information of the considered routing river stretch without considering lateral flow are estimated using the Robust Parameter Estimation (ROPE) algorithm that allows for arriving at the best performing set of bed width and side slope of a trapezoidal section. The performance of the VPMM method is evaluated using the Nash-Sutcliffe model efficiency criterion as the objective function to be maximized using the ROPE algorithm. The twenty-seven flood events in the calibration set are considered to identify the relationship between 'total rainfall' and 'total losses' as well as to optimize the geometric characteristics of the prismatic channel (width and slope of the trapezoidal section). Based on this analysis, a relationship between total rainfall and total loss of the intermediate catchment is obtained and then used to estimate the lateral flow in the reach. Assuming the lateral flow hydrograph is of the form of inflow hydrograph and using the total intervening catchment runoff estimated from the relationship, the uniformly distributed lateral flow rate qL at any instant of time is estimated for its use in the VPMM routing method. All the 27 flood events are simulated using this routing approach considering lateral flow along the reach. Many of these simulations are able to simulate the observed hydrographs very closely. The proposed approach

  9. Analysis of lignans in Magnoliae Flos by turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry.

    PubMed

    Zhou, Xuan; Chen, Cen; Ye, Xiaolan; Song, Fenyun; Fan, Guorong; Wu, Fuhai

    2016-04-01

    In this study, a method coupling turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry was developed for analyzing the lignans in Magnoliae Flos. By the online pretreatment of turbulent flow chromatography solid-phase extraction, the impurities removal and analytes concentration were automatically processed, and the lignans were separated rapidly and well. Seven lignans of Magnoliae Flos including epieudesmin, magnolin, 1-irioresinol-B-dimethyl ether, epi-magnolin, fargesin aschantin, and demethoxyaschantin were identified by comparing their retention behavior, UV spectra, and mass spectra with those of reference substances or literature data. The developed method was validated, and the good results showed that the method was not only automatic and rapid, but also accurate and reliable. The turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry method holds a high potential to become an effective method for the quality control of lignans in Magnoliae Flos and a useful tool for the analysis of other complex mixtures. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Replacing antibodies with aptamers in lateral flow immunoassay.

    PubMed

    Chen, Ailiang; Yang, Shuming

    2015-09-15

    Aptamers have been identified against various targets as a type of chemical or nucleic acid ligand by systematic evolution of ligands by exponential enrichment (SELEX) with high sensitivity and specificity. Aptamers show remarkable advantages over antibodies due to the nucleic acid nature and target-induced structure-switching properties and are widely used to design various fluorescent, electrochemical, or colorimetric biosensors. However, the practical applications of aptamer-based sensing and diagnostics are still lagging behind those of antibody-based tests. Lateral flow immunoassay (LFIA) represents a well established and appropriate technology among rapid assays because of its low cost and user-friendliness. The antibody-based platform is utilized to detect numerous targets, but it is always hampered by the antibody preparation time, antibody stability, and effect of modification on the antibody. Seeking alternatives to antibodies is an area of active research and is of tremendous importance. Aptamers are receiving increasing attention in lateral flow applications because of a number of important potential performance advantages. We speculate that aptamer-based LFIA may be one of the first platforms for commercial use of aptamer-based diagnosis. This review first gives an introduction to aptamer including the selection process SELEX with its focus on aptamer advantages over antibodies, and then depicts LFIA with its focus on aptamer opportunities in LFIA over antibodies. Furthermore, we summarize the recent advances in the development of aptamer-based lateral flow biosensing assays with the aim to provide a general guide for the design of aptamer-based lateral flow biosensing assays. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Modeling lateral circulation and its influence on the along-channel flow in a branched estuary

    NASA Astrophysics Data System (ADS)

    Zhu, Lei; He, Qing; Shen, Jian

    2018-02-01

    A numerical modeling study of the influence of the lateral flow on the estuarine exchange flow was conducted in the north passage of the Changjiang estuary. The lateral flows show substantial variabilities within a flood-ebb tidal cycle. The strong lateral flow occurring during flood tide is caused primarily by the unique cross-shoal flow that induces a strong northward (looking upstream) barotropic force near the surface and advects saltier water toward the northern part of the channel, resulting in a southward baroclinic force caused by the lateral density gradient. Thus, a two-layer structure of lateral flows is produced during the flood tide. The lateral flows are vigorous near the flood slack and the magnitude can exceed that of the along-channel tidal flow during that period. The strong vertical shear of the lateral flows and the salinity gradient in lateral direction generate lateral tidal straining, which are out of phase with the along-channel tidal straining. Consequently, stratification is enhanced at the early stage of the ebb tide. In contrast, strong along-channel straining is apparent during the late ebb tide. The vertical mixing disrupts the vertical density gradient, thus suppressing stratification. The impact of lateral straining on stratification during spring tide is more pronounced than that of along-channel straining during late flood and early ebb tides. The momentum balance along the estuary suggests that lateral flow can augment the residual exchange flow. The advection of lateral flows brings low-energy water from the shoal to the deep channel during the flood tide, whereas the energetic water is moved to the shoal via lateral advection during the ebb tide. The impact of lateral flow on estuarine circulation of this multiple-channel estuary is different from single-channel estuary. A model simulation by blocking the cross-shoal flow shows that the magnitudes of lateral flows and tidal straining are reduced. Moreover, the reduced lateral

  12. Rapid preparative separation of monoclonal antibody charge variants using laterally-fed membrane chromatography.

    PubMed

    Sadavarte, Rahul; Madadkar, Pedram; Filipe, Carlos Dm; Ghosh, Raja

    2018-01-15

    Monoclonal antibodies undergo various forms of chemical transformation which have been shown to cause loss in efficacy and alteration in pharmacokinetic properties of these molecules. Such modified antibody molecules are known as variants. They also display physical properties such as charge that are different from intact antibody molecules. However, the difference in charge is very subtle and separation based on it is quite challenging. Charge variants are usually separated using ion-exchange column chromatography or isoelectric focusing. In this paper, we report a rapid and scalable method for fractionating monoclonal antibody charge variants, based on the use of cation exchange laterally-fed membrane chromatography (LFMC). Starting with a sample of monoclonal antibody hIgG1-CD4, three well-resolved fractions were obtained using either pH or salt gradient. These fractions were identified as acidic, neutral and basic variants. Each of these fractions contained intact heavy and light chains and so antibody fragmentation had no role in variant generation. The separation was comparable to that using column chromatography but was an order of magnitude faster. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Research on Flow Field Perception Based on Artificial Lateral Line Sensor System

    PubMed Central

    Wang, Anyi; Wang, Shirui; Yang, Tingting

    2018-01-01

    In nature, the lateral line of fish is a peculiar and important organ for sensing the surrounding hydrodynamic environment, preying, escaping from predators and schooling. In this paper, by imitating the mechanism of fish lateral canal neuromasts, we developed an artificial lateral line system composed of micro-pressure sensors. Through hydrodynamic simulations, an optimized sensor structure was obtained and the pressure distribution models of the lateral surface were established in uniform flow and turbulent flow. Carrying out the corresponding underwater experiment, the validity of the numerical simulation method is verified by the comparison between the experimental data and the simulation results. In addition, a variety of effective research methods are proposed and validated for the flow velocity estimation and attitude perception in turbulent flow, respectively and the shape recognition of obstacles is realized by the neural network algorithm. PMID:29534499

  14. Research on Flow Field Perception Based on Artificial Lateral Line Sensor System.

    PubMed

    Liu, Guijie; Wang, Mengmeng; Wang, Anyi; Wang, Shirui; Yang, Tingting; Malekian, Reza; Li, Zhixiong

    2018-03-11

    In nature, the lateral line of fish is a peculiar and important organ for sensing the surrounding hydrodynamic environment, preying, escaping from predators and schooling. In this paper, by imitating the mechanism of fish lateral canal neuromasts, we developed an artificial lateral line system composed of micro-pressure sensors. Through hydrodynamic simulations, an optimized sensor structure was obtained and the pressure distribution models of the lateral surface were established in uniform flow and turbulent flow. Carrying out the corresponding underwater experiment, the validity of the numerical simulation method is verified by the comparison between the experimental data and the simulation results. In addition, a variety of effective research methods are proposed and validated for the flow velocity estimation and attitude perception in turbulent flow, respectively and the shape recognition of obstacles is realized by the neural network algorithm.

  15. Imaging dipole flow sources using an artificial lateral-line system made of biomimetic hair flow sensors

    PubMed Central

    Dagamseh, Ahmad; Wiegerink, Remco; Lammerink, Theo; Krijnen, Gijs

    2013-01-01

    In Nature, fish have the ability to localize prey, school, navigate, etc., using the lateral-line organ. Artificial hair flow sensors arranged in a linear array shape (inspired by the lateral-line system (LSS) in fish) have been applied to measure airflow patterns at the sensor positions. Here, we take advantage of both biomimetic artificial hair-based flow sensors arranged as LSS and beamforming techniques to demonstrate dipole-source localization in air. Modelling and measurement results show the artificial lateral-line ability to image the position of dipole sources accurately with estimation error of less than 0.14 times the array length. This opens up possibilities for flow-based, near-field environment mapping that can be beneficial to, for example, biologists and robot guidance applications. PMID:23594816

  16. High-resolution, preparative purification of PEGylated protein using a laterally-fed membrane chromatography device.

    PubMed

    Madadkar, Pedram; Nino, Sergio Luna; Ghosh, Raja

    2016-11-01

    We discuss the use of a laterally-fed membrane chromatography (or LFMC) device for single-step purification of mono-PEGylated lysozyme. Recent studies have shown such LFMC devices to be suitable for high-resolution, multi-component separation of proteins in the bind-and-elute mode. The device used in this study contained a stack of rectangular cation-exchange membranes having 9.25mL bed volume. PEGylation of lysozyme was carried out in batch mode using 5kDa methoxy-polyethyleneglycol propionaldehyde (or m-PEG propionaldehyde) in the presence of sodium cyanoborohydride as reducing agent. Membrane chromatographic separation was carried out at 1.62 membrane bed volumes per minute flow rate, in the bind-and-elute mode. When a salt gradient was applied, the higher PEGylated forms of lysozyme (i.e. the byproducts) eluted earlier than mono-PEGylated lysozyme (the target product), while lysozyme eluted last. Under elution conditions optimized for resolution and speed, the separation could be carried out in less than 15 membrane bed volumes. High purity and recovery of mono-PEGylated lysozyme was obtained. The resolution of separation of mono-PEGylated lysozyme obtained under the above condition was comparable to that reported in the literature for equivalent cation-exchange resin columns while the flow rate expressed in bed volumes/min was 21.7 times higher. Also, the number of theoretical plates per meter was significantly higher with the LFMC device. Therefore the LFMC based purification process discussed in this paper combined high-productivity with high-resolution. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Species Specific Bacterial Spore Detection Using Lateral-Flow Immunoassay with DPA-Triggered Tb Luminescence

    NASA Technical Reports Server (NTRS)

    Ponce, Adrian

    2003-01-01

    A method of detecting bacterial spores incorporates (1) A method of lateral-flow immunoassay in combination with (2) A method based on the luminescence of Tb3+ ions to which molecules of dipicolinic acid (DPA) released from the spores have become bound. The present combination of lateral-flow immunoassay and DPA-triggered Tb luminescence was developed as a superior alternative to a prior lateral-flow immunoassay method in which detection involves the visual observation and/or measurement of red light scattered from colloidal gold nanoparticles. The advantage of the present combination method is that it affords both (1) High selectivity for spores of the species of bacteria that one seeks to detect (a characteristic of lateral-flow immunoassay in general) and (2) Detection sensitivity much greater (by virtue of the use of DPA-triggered Tb luminescence instead of gold nanoparticles) than that of the prior lateral-flow immunoassay method

  18. High frequency lateral flow affinity assay using superparamagnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Lago-Cachón, D.; Rivas, M.; Martínez-García, J. C.; Oliveira-Rodríguez, M.; Blanco-López, M. C.; García, J. A.

    2017-02-01

    Lateral flow assay is one of the simplest and most extended techniques in medical diagnosis for point-of-care testing. Although it has been traditionally a positive/negative test, some work has been lately done to add quantitative abilities to lateral flow assay. One of the most successful strategies involves magnetic beads and magnetic sensors. Recently, a new technique of superparamagnetic nanoparticle detection has been reported, based on the increase of the impedance induced by the nanoparticles on a RF-current carrying copper conductor. This method requires no external magnetic field, which reduces the system complexity. In this work, nitrocellulose membranes have been installed on the sensor, and impedance measurements have been carried out during the sample diffusion by capillarity along the membrane. The impedance of the sensor changes because of the presence of magnetic nanoparticles. The results prove the potentiality of the method for point-of-care testing of biochemical substances and nanoparticle capillarity flow studies.

  19. A Lateral Flow Biosensor for the Detection of Single Nucleotide Polymorphisms.

    PubMed

    Zeng, Lingwen; Xiao, Zhuo

    2017-01-01

    A lateral flow biosensor (LFB) is introduced for the detection of single nucleotide polymorphisms (SNPs). The assay is composed of two steps: circular strand displacement reaction and lateral flow biosensor detection. In step 1, the nucleotide at SNP site is recognized by T4 DNA ligase and the signal is amplified by strand displacement DNA polymerase, which can be accomplished at a constant temperature. In step 2, the reaction product of step 1 is detected by a lateral flow biosensor, which is a rapid and cost effective tool for nuclei acid detection. Comparing with conventional methods, it requires no complicated machines. It is suitable for the use of point of care diagnostics. Therefore, this simple, cost effective, robust, and promising LFB detection method of SNP has great potential for the detection of genetic diseases, personalized medicine, cancer related mutations, and drug-resistant mutations of infectious agents.

  20. Liquid chromatography/Fourier transform IR spectrometry interface flow cell

    DOEpatents

    Johnson, Charles C.; Taylor, Larry T.

    1986-01-01

    A zero dead volume (ZDV) microbore high performance liquid chromatography (.mu.HPLC)/Fourier transform infrared (FTIR) interface flow cell includes an IR transparent crystal having a small diameter bore therein through which a sample liquid is passed. The interface flow cell further includes a metal holder in combination with a pair of inner, compressible seals for directly coupling the thus configured spectrometric flow cell to the outlet of a .mu.HPLC column end fitting to minimize the transfer volume of the effluents exiting the .mu.HPLC column which exhibit excellent flow characteristics due to the essentially unencumbered, open-flow design. The IR beam passes transverse to the sample flow through the circular bore within the IR transparent crystal, which is preferably comprised of potassium bromide (KBr) or calcium fluoride (CaF.sub.2), so as to minimize interference patterns and vignetting encountered in conventional parallel-plate IR cells. The long IR beam pathlength and lensing effect of the circular cross-section of the sample volume in combination with the refractive index differences between the solvent and the transparent crystal serve to focus the IR beam in enhancing sample detection sensitivity by an order of magnitude.

  1. Liquid chromatography/Fourier transform IR spectrometry interface flow cell

    DOEpatents

    Johnson, C.C.; Taylor, L.T.

    1985-01-04

    A zero dead volume (ZDV) microbore high performance liquid chromatography (..mu.. HPLC)/Fourier transform infrared (FTIR) interface flow cell includes an IR transparent crystal having a small diameter bore therein through which a sample liquid is passed. The interface flow cell further includes a metal holder in combination with a pair of inner, compressible seals for directly coupling the thus configured spectrometric flow cell to the outlet of a ..mu.. HPLC column end fitting to minimize the transfer volume of the effluents exiting the ..mu.. HPLC column which exhibit excellent flow characteristics due to the essentially unencumbered, open-flow design. The IR beam passes transverse to the sample flow through the circular bore within the IR transparent crystal, which is preferably comprised of potassium bromide (KBr) or calcium fluoride (CaF/sub 2/), so as to minimize interference patterns and vignetting encountered in conventional parallel-plate IR cells. The long IR beam pathlength and lensing effect of the circular cross-section of the sample volume in combination with the refractive index differences between the solvent and the transparent crystal serve to focus the IR beam in enhancing sample detection sensitivity by an order of magnitude.

  2. Quantifiable Lateral Flow Assay Test Strips

    NASA Technical Reports Server (NTRS)

    2003-01-01

    As easy to read as a home pregnancy test, three Quantifiable Lateral Flow Assay (QLFA) strips used to test water for E. coli show different results. The brightly glowing control line on the far right of each strip indicates that all three tests ran successfully. But the glowing test line on the middle left and bottom strips reveal their samples were contaminated with E. coli bacteria at two different concentrations. The color intensity correlates with concentration of contamination.

  3. Multiphase flow modeling in centrifugal partition chromatography.

    PubMed

    Adelmann, S; Schwienheer, C; Schembecker, G

    2011-09-09

    The separation efficiency in Centrifugal Partition Chromatography (CPC) depends on selection of a suitable biphasic solvent system (distribution ratio, selectivity factor, sample solubility) and is influenced by hydrodynamics in the chambers. Especially the stationary phase retention, the interfacial area for mass transfer and the flow pattern (backmixing) are important parameters. Their relationship with physical properties, operating parameters and chamber geometry is not completely understood and predictions are hardly possible. Experimental flow visualization is expensive and two-dimensional only. Therefore we simulated the flow pattern using a volume-of-fluid (VOF) method, which was implemented in OpenFOAM®. For the three-dimensional simulation of a rotating FCPC®-chamber, gravitational centrifugal and Coriolis forces were added to the conservation equation. For experimental validation the flow pattern of different solvent systems was visualized with an optical measurement system. The amount of mobile phase in a chamber was calculated from gray scale values of videos recorded by an image processing routine in ImageJ®. To visualize the flow of the stationary phase polyethylene particles were used to perform a qualitative particle image velocimetry (PIV) analysis. We found a good agreement between flow patterns and velocity profiles of experiments and simulations. By using the model we found that increasing the chamber depth leads to higher specific interfacial area. Additionally a circular flow in the stationary phase was identified that lowers the interfacial area because it pushes the jet of mobile phase to the chamber wall. The Coriolis force alone gives the impulse for this behavior. As a result the model is easier to handle than experiments and allows 3D prediction of hydrodynamics in the chamber. Additionally it can be used for optimizing geometry and operating parameters for given physical properties of solvent systems. Copyright © 2011 Elsevier B

  4. Comparison of Lateral Flow Assay, Kidney Inhibition Swab, and Liquid Chromatography-Tandem Mass Spectrometry for the Detection of Penicillin G Residues in Sow Urine.

    PubMed

    Shelver, Weilin L; Chakrabarty, Shubhashis; Smith, David J

    2017-03-01

    Sows (n = 126) were administered penicillin G; urine, collected at slaughter, was screened by kidney inhibition swab (KIS; 4 h testing time) and then stored at -80 °C (∼1200 days) until analysis by lateral flow assay (LF, ∼5 min testing time) and tandem quadrupole LC-MS/MS (TQ) analysis. The stability of penicillin in urine during storage was verified using TQ analyses. Quantitative results were well-correlated (R 2 = 0.98) with only a ∼10% decrease in penicillin concentration during the 3-year storage period. KIS retesting of stored samples returned results consistent with the original analyses. Lateral flow assay results were highly correlated with the KIS and TQ results. A KIS positive sample, which was not confirmed by TQ or LF, was assayed by Triple-TOF LC-MS to determine the cause of the apparent false positive. This study suggests LF can be used to quickly and efficiently screen for penicillin G residues before slaughter.

  5. Aptamer-phage reporters for ultrasensitive lateral flow assays

    PubMed Central

    Adhikari, Meena; Strych, Ulrich; Kim, Jinsu; Goux, Heather; Dhamane, Sagar; Poongavanam, Mohan-Vivekanandan; Hagström, Anna E. V.; Kourentzi, Katerina; Conrad, Jacinta C.; Willson, Richard C.

    2015-01-01

    We introduce the modification of bacteriophage particles with aptamers for the use as bioanalytical reporters, and demonstrate the use of these particles in ultrasensitive lateral flow assays. M13 phage displaying an in vivo biotinylatable peptide (AviTag) genetically fused to the phage tail protein pIII were used as reporter particle scaffolds, with biotinylated aptamers attached via avidin-biotin linkages, and horseradish peroxidase (HRP) reporter enzymes covalently attached to the pVIII coat protein. These modified viral nanoparticles were used in immunochromatographic sandwich assays for the direct detection of IgE and of the penicillin-binding protein from Staphylococcus aureus (PBP2a). We also developed an additional lateral flow assay for IgE, in which the analyte is sandwiched between immobilized anti-IgE antibodies and aptamer-bearing reporter phage modified with HRP. The limit of detection of this LFA was 0.13 ng/mL IgE, ~100 times lower than those of previously reported IgE assays. PMID:26456715

  6. Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays.

    PubMed

    Adhikari, Meena; Strych, Ulrich; Kim, Jinsu; Goux, Heather; Dhamane, Sagar; Poongavanam, Mohan-Vivekanandan; Hagström, Anna E V; Kourentzi, Katerina; Conrad, Jacinta C; Willson, Richard C

    2015-12-01

    We introduce the modification of bacteriophage particles with aptamers for use as bioanalytical reporters, and demonstrate the use of these particles in ultrasensitive lateral flow assays. M13 phage displaying an in vivo biotinylatable peptide (AviTag) genetically fused to the phage tail protein pIII were used as reporter particle scaffolds, with biotinylated aptamers attached via avidin-biotin linkages, and horseradish peroxidase (HRP) reporter enzymes covalently attached to the pVIII coat protein. These modified viral nanoparticles were used in immunochromatographic sandwich assays for the direct detection of IgE and of the penicillin-binding protein from Staphylococcus aureus (PBP2a). We also developed an additional lateral flow assay for IgE, in which the analyte is sandwiched between immobilized anti-IgE antibodies and aptamer-bearing reporter phage modified with HRP. The limit of detection of this LFA was 0.13 ng/mL IgE, ∼100 times lower than those of previously reported IgE assays.

  7. Flow-switching device for comprehensive two-dimensional gas chromatography.

    PubMed

    Bueno, Pedro A; Seeley, John V

    2004-02-20

    A simple flow-switching device has been developed as a differential flow modulator for comprehensive two-dimensional gas chromatography (GC x GC). The device is assembled from tubing, four tee unions, and a solenoid valve. The solenoid valve is located outside the oven of the gas chromatograph and is not in the sample path. The modulation technique has no inherent temperature restrictions and passes 100% of the primary column effluent to the secondary column(s). Secondary peaks are produced with widths at half maximum less than 100 ms when operating in GC x 2GC mode with a 2.0 s modulation period. The efficacy of this approach is demonstrated through the analysis of a standard mixture of volatile organic compounds (VOCs) and diesel fuel.

  8. Multiplexed lateral flow biosensors: Technological advances for radically improving point-of-care diagnoses.

    PubMed

    Li, Jia; Macdonald, Joanne

    2016-09-15

    Lateral flow biosensors are a leading technology in point-of-care diagnostics due to their simplicity, rapidness and low cost. Their primacy in this arena continues through technological breakthroughs such as multiplexing: the detection of more than one biomarker in a single assay. Multiplexing capacity is critical for improving diagnostic efficiency, enhancing the diagnostic precision for specific diseases and reducing diagnostic cost. Here we review, for the first time, the various types and strategies employed for creating multiplexed lateral flow biosensors. These are classified into four main categories in terms of specific application or multiplexing level, namely linear, parameter, spatial and conceptual. We describe the practical applications and implications for each approach and compare their advantages and disadvantages. Importantly, multiplexing is still subject to limitations of the traditional lateral flow biosensor, such as sensitivity and specificity. However, by pushing the limitations of the traditional medium into the multiplex arena, several technological breakthroughs are emerging with novel solutions that further expand the utility of lateral flow biosensing for point-of-care applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Occurrence of turbulent flow conditions in supercritical fluid chromatography.

    PubMed

    De Pauw, Ruben; Choikhet, Konstantin; Desmet, Gert; Broeckhoven, Ken

    2014-09-26

    Having similar densities as liquids but with viscosities up to 20 times lower (higher diffusion coefficients), supercritical CO2 is the ideal (co-)solvent for fast and/or highly efficient separations without mass-transfer limitations or excessive column pressure drops. Whereas in liquid chromatography the flow remains laminar in both the packed bed and tubing, except in extreme cases (e.g. in a 75 μm tubing, pure acetonitrile at 5 ml/min), a supercritical fluid can experience a transition from laminar to turbulent flow in more typical operation modes. Due to the significant lower viscosity, this transition for example already occurs at 1.3 ml/min for neat CO2 when using connection tubing with an ID of 127 μm. By calculating the Darcy friction factor, which can be plotted versus the Reynolds number in a so-called Moody chart, typically used in fluid dynamics, higher values are found for stainless steel than PEEK tubing, in agreement with their expected higher surface roughness. As a result turbulent effects are more pronounced when using stainless steel tubing. The higher than expected extra-column pressure drop limits the kinetic performance of supercritical fluid chromatography and complicates the optimization of tubing ID, which is based on a trade-off between extra-column band broadening and pressure drop. One of the most important practical consequences is the non-linear increase in extra-column pressure drop over the tubing downstream of the column which leads to an unexpected increase in average column pressure and mobile phase density, and thus decrease in retention. For close eluting components with a significantly different dependence of retention on density, the selectivity can significantly be affected by this increase in average pressure. In addition, the occurrence of turbulent flow is also observed in the detector cell and connection tubing. This results in a noise-increase by a factor of four when going from laminar to turbulent flow (e.g. going

  10. Lateral fluid flow in a compacting sand-shale sequence: South Caspian basin.

    USGS Publications Warehouse

    Bredehoeft, J.D.; Djevanshir, R.D.; Belitz, K.R.

    1988-01-01

    The South Caspian basin contains both sands and shales that have pore-fluid pressures substantially in excess of hydrostatic fluid pressure. Pore-pressure data from the South Caspian basin demonstrate that large differences in excess hydraulic head exist between sand and shale. The data indicate that sands are acting as drains for overlying and underlying compacting shales and that fluid flows laterally through the sand on a regional scale from the basin interior northward to points of discharge. The major driving force for the fluid movement is shale compaction. We present a first- order mathematical analysis in an effort to test if the permeability of the sands required to support a regional flow system is reasonable. The results of the analysis suggest regional sand permeabilities ranging from 1 to 30 md; a range that seems reasonable. This result supports the thesis that lateral fluid flow is occurring on a regional scale within the South Caspian basin. If vertical conduits for flow exist within the basin, they are sufficiently impermeable and do not provide a major outlet for the regional flow system. The lateral fluid flow within the sands implies that the stratigraphic sequence is divided into horizontal units that are hydraulically isolated from one another, a conclusion that has important implications for oil and gas migration.-Authors

  11. Analytical Tools to Improve Optimization Procedures for Lateral Flow Assays

    PubMed Central

    Hsieh, Helen V.; Dantzler, Jeffrey L.; Weigl, Bernhard H.

    2017-01-01

    Immunochromatographic or lateral flow assays (LFAs) are inexpensive, easy to use, point-of-care medical diagnostic tests that are found in arenas ranging from a doctor’s office in Manhattan to a rural medical clinic in low resource settings. The simplicity in the LFA itself belies the complex task of optimization required to make the test sensitive, rapid and easy to use. Currently, the manufacturers develop LFAs by empirical optimization of material components (e.g., analytical membranes, conjugate pads and sample pads), biological reagents (e.g., antibodies, blocking reagents and buffers) and the design of delivery geometry. In this paper, we will review conventional optimization and then focus on the latter and outline analytical tools, such as dynamic light scattering and optical biosensors, as well as methods, such as microfluidic flow design and mechanistic models. We are applying these tools to find non-obvious optima of lateral flow assays for improved sensitivity, specificity and manufacturing robustness. PMID:28555034

  12. Investigation of particle lateral migration in sample-sheath flow of viscoelastic fluid and Newtonian fluid.

    PubMed

    Yuan, Dan; Zhang, Jun; Yan, Sheng; Peng, Gangrou; Zhao, Qianbin; Alici, Gursel; Du, Hejun; Li, Weihua

    2016-08-01

    In this work, particle lateral migration in sample-sheath flow of viscoelastic fluid and Newtonian fluid was experimentally investigated. The 4.8-μm micro-particles were dispersed in a polyethylene oxide (PEO) viscoelastic solution, and then the solution was injected into a straight rectangular channel with a deionised (DI) water Newtonian sheath flow. Micro-particles suspended in PEO solution migrated laterally to a DI water stream, but migration in the opposite direction from a DI water stream to a PEO solution stream or from one DI water stream to another DI water stream could not be achieved. The lateral migration of particles depends on the viscoelastic properties of the sample fluids. Furthermore, the effects of channel length, flow rate, and PEO concentration were studied. By using viscoelastic sample flow and Newtonian sheath flow, a selective particle lateral migration can be achieved in a simple straight channel, without any external force fields. This particle lateral migration technique could be potentially used in solution exchange fields such as automated cell staining and washing in microfluidic platforms, and holds numerous biomedical applications. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Flow visualization of lateral jet injection into swirling crossflow

    NASA Technical Reports Server (NTRS)

    Ferrell, G. B.; Aoki, K.; Lilley, D. G.

    1985-01-01

    Flow visualization experiments have been conducted to characterize the time-mean flowfield of a deflected turbulent jet in a confining cylindrical crossflow. Jet-to-crossflow velocity ratios of 2, 4, and 6 were investigated, under crossflow inlet swirler vane angles of 0 (swirler removed), 45 and 70 degrees. Smoke, neutrally-buoyant helium-filled soap bubbles, and multi-spark flow visualization were employed to highlight interesting features of the deflected jet, as well as the trajectory and spread pattern of the jet. Gross flowfield characterization was obtained for a range of lateral jet-to-crossflow velocity ratios and a range of inlet swirl strengths in the main flow. The flow visualization results agree well with the measurements obtained elsewhere with the six-orientation single hot-wire method.

  14. Wave propagation and power flow in an acoustic metamaterial plate with lateral local resonance attachment

    NASA Astrophysics Data System (ADS)

    Wang, Ting; Sheng, Meiping; Ding, Xiaodong; Yan, Xiaowei

    2018-03-01

    This paper presents analysis on wave propagation and power flow in an acoustic metamaterial plate with lateral local resonance. The metamaterial is designed to have lateral local resonance systems attached to a homogeneous plate. Relevant theoretical analysis, numerical modelling and application prospect are presented. Results show that the metamaterial has two complete band gaps for flexural wave absorption and vibration attenuation. Damping can smooth and lower the metamaterial’s frequency responses in high frequency ranges at the expense of the band gap effect, and as an important factor to calculate the power flow is thoroughly investigated. Moreover, the effective mass density becomes negative and unbounded at specific frequencies. Simultaneously, power flow within band gaps are dramatically blocked from the power flow contour and power flow maps. Results from finite element modelling and power flow analysis reveal the working mechanism of the flexural wave attenuation and power flow blocked within the band gaps, where part of the flexural vibration is absorbed by the vertical resonator and the rest is transformed through four-link-mechanisms to the lateral resonators that oscillate and generate inertial forces indirectly to counterbalance the shear forces induced by the vibrational plate. The power flow is stored in the vertical and lateral local resonance, as well as in the connected plate.

  15. Additional band broadening of peptides in the first size-exclusion chromatographic dimension of an automated stop-flow two-dimensional high performance liquid chromatography.

    PubMed

    Xu, Jucai; Sun-Waterhouse, Dongxiao; Qiu, Chaoying; Zhao, Mouming; Sun, Baoguo; Lin, Lianzhu; Su, Guowan

    2017-10-27

    The need to improve the peak capacity of liquid chromatography motivates the development of two-dimensional analysis systems. This paper presented a fully automated stop-flow two-dimensional liquid chromatography system with size exclusion chromatography followed by reversed phase liquid chromatography (SEC×RPLC) to efficiently separate peptides. The effects of different stop-flow operational parameters (stop-flow time, peak parking position, number of stop-flow periods and column temperature) on band broadening in the first dimension (1 st D) SEC column were quantitatively evaluated by using commercial small proteins and peptides. Results showed that the effects of peak parking position and the number of stop-flow periods on band broadening were relatively small. Unlike stop-flow analysis of large molecules with a long running time, additional band broadening was evidently observed for small molecule analytes due to the relatively high effective diffusion coefficient (D eff ). Therefore, shorter analysis time and lower 1 st D column temperature were suggested for analyzing small molecules. The stop-flow two-dimensional liquid chromatography (2D-LC) system was further tested on peanut peptides and an evidently improved resolution was observed for both stop-flow heart-cutting and comprehensive 2D-LC analysis (in spite of additional band broadening in SEC). The stop-flow SEC×RPLC, especially heart-cutting analysis with shorter analysis time and higher 1 st D resolution for selected fractions, offers a promising approach for efficient analysis of complex samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Simulation and validation of concentrated subsurface lateral flow paths in an agricultural landscape

    NASA Astrophysics Data System (ADS)

    Zhu, Q.; Lin, H. S.

    2009-08-01

    The importance of soil water flow paths to the transport of nutrients and contaminants has long been recognized. However, effective means of detecting concentrated subsurface flow paths in a large landscape are still lacking. The flow direction and accumulation algorithm based on single-direction flow algorithm (D8) in GIS hydrologic modeling is a cost-effective way to simulate potential concentrated flow paths over a large area once relevant data are collected. This study tested the D8 algorithm for simulating concentrated lateral flow paths at three interfaces in soil profiles in a 19.5-ha agricultural landscape in central Pennsylvania, USA. These interfaces were (1) the interface between surface plowed layers of Ap1 and Ap2 horizons, (2) the interface with subsoil water-restricting clay layer where clay content increased to over 40%, and (3) the soil-bedrock interface. The simulated flow paths were validated through soil hydrologic monitoring, geophysical surveys, and observable soil morphological features. The results confirmed that concentrated subsurface lateral flow occurred at the interfaces with the clay layer and the underlying bedrock. At these two interfaces, the soils on the simulated flow paths were closer to saturation and showed more temporally unstable moisture dynamics than those off the simulated flow paths. Apparent electrical conductivity in the soil on the simulated flow paths was elevated and temporally unstable as compared to those outside the simulated paths. The soil cores collected from the simulated flow paths showed significantly higher Mn content at these interfaces than those away from the simulated paths. These results suggest that (1) the D8 algorithm is useful in simulating possible concentrated subsurface lateral flow paths if used with appropriate threshold value of contributing area and sufficiently detailed digital elevation model (DEM); (2) repeated electromagnetic surveys can reflect the temporal change of soil water storage

  17. Compressible Flow Phenomena at Inception of Lateral Density Currents Fed by Collapsing Gas-Particle Mixtures

    NASA Astrophysics Data System (ADS)

    Valentine, Greg A.; Sweeney, Matthew R.

    2018-02-01

    Many geological flows are sourced by falling gas-particle mixtures, such as during collapse of lava domes, and impulsive eruptive jets, and sustained columns, and rock falls. The transition from vertical to lateral flow is complex due to the range of coupling between particles of different sizes and densities and the carrier gas, and due to the potential for compressible flow phenomena. We use multiphase modeling to explore these dynamics. In mixtures with small particles, and with subsonic speeds, particles follow the gas such that outgoing lateral flows have similar particle concentration and speed as the vertical flows. Large particles concentrate immediately upon impact and move laterally away as granular flows overridden by a high-speed jet of expelled gas. When a falling flow is supersonic, a bow shock develops above the impact zone, and this produces a zone of high pressure from which lateral flows emerge as overpressured wall jets. The jets form complex structures as the mixtures expand and accelerate and then recompress through a recompression zone that mimics a Mach disk shock in ideal gas jets. In mixtures with moderate to high ratios of fine to coarse particles, the latter tend to follow fine particles through the expansion-recompression flow fields because of particle-particle drag. Expansion within the flow fields can lead to locally reduced gas pressure that could enhance substrate erosion in natural flows. The recompression zones form at distances, and have peak pressures, that are roughly proportional to the Mach numbers of impacting flows.

  18. A silicon dioxide modified magnetic nanoparticles-labeled lateral flow strips for HBs antigen.

    PubMed

    Zhang, Xueqing; Jiang, Lin; Zhang, Chunlei; Li, Ding; Wang, Can; Gao, Feng; Cui, Daxiang

    2011-12-01

    Herein we reported a new type of silicon dioxide wrapped magnetic nanoparticles-labeled lateral flow strip for detection of HBs antigen in sera. The SiO2 wrapped Fe3O4 nanocomposites were prepared and characterized by HR-TEM, FTIR and magnetometer. As-prepared nanocomposites were used to label anti-HBV surface monoclonal antibody, the lateral flow strips were constructed, and 100 specimens of sera were collected and tested. Results showed that the prepared SiO2 wrapped Fe3O4 nanocomposites were shell/core structure, well dispersed, with the size of 25 nm in diameter, the thickness of the shell was about 3 nm, their magnetic saturation intensity was 44.3 meu g(-1). Clinical sera specimens test results showed that the prepared lateral flow strips were with the detection limitation of 5 pg/mL by naked eye observation, and 0.1 pg/mL by CCD reader or MAR Analyzer, specificity was 100%. In conclusion, one kind of silicon dioxide wrapped magnetic nanoparticles-labeled lateral flow strip for ultrasensitive detection of HBs antigen was successfully developed, its ease of use, sensitiveness and low-cost make it well-suited for population-based on-the-site hepatitis B screening.

  19. New Fpg probe chemistry for direct detection of recombinase polymerase amplification on lateral flow strips.

    PubMed

    Powell, Michael L; Bowler, Frank R; Martinez, Aurore J; Greenwood, Catherine J; Armes, Niall; Piepenburg, Olaf

    2018-02-15

    Rapid, cost-effective and sensitive detection of nucleic acids has the ability to improve upon current practices employed for pathogen detection in diagnosis of infectious disease and food testing. Furthermore, if assay complexity can be reduced, nucleic acid amplification tests could be deployed in resource-limited and home use scenarios. In this study, we developed a novel Fpg (Formamidopyrimidine DNA glycosylase) probe chemistry, which allows lateral flow detection of amplification in undiluted recombinase polymerase amplification (RPA) reactions. The prototype nucleic acid lateral flow chemistry was applied to a human genomic target (rs1207445), Campylobacter jejuni 16S rDNA and two genetic markers of the important food pathogen E. coli O157:H7. All four assays have an analytical sensitivity between 10 and 100 copies DNA per amplification. Furthermore, the assay is performed with fewer hands-on steps than using the current RPA Nfo lateral flow method as dilution of amplicon is not required for lateral flow analysis. Due to the simplicity of the workflow, we believe that the lateral flow chemistry for direct detection could be readily adapted to a cost-effective single-use consumable, ideal for use in non-laboratory settings. Copyright © 2017. Published by Elsevier Inc.

  20. An Inexpensive, Fast and Sensitive Quantitative Lateral Flow Magneto-Immunoassay for Total Prostate Specific Antigen

    PubMed Central

    Barnett, Jacqueline M.; Wraith, Patrick; Kiely, Janice; Persad, Raj; Hurley, Katrina; Hawkins, Peter; Luxton, Richard

    2014-01-01

    We describe the detection characteristics of a device the Resonant Coil Magnetometer (RCM) to quantify paramagnetic particles (PMPs) in immunochromatographic (lateral flow) assays. Lateral flow assays were developed using PMPs for the measurement of total prostate specific antigen (PSA) in serum samples. A detection limit of 0.8 ng/mL was achieved for total PSA using the RCM and is at clinically significant concentrations. Comparison of data obtained in a pilot study from the analysis of serum samples with commercially available immunoassays shows good agreement. The development of a quantitative magneto-immunoassay in lateral flow format for total PSA suggests the potential of the RCM to operate with many immunoassay formats. The RCM has the potential to be modified to quantify multiple analytes in this format. This research shows promise for the development of an inexpensive device capable of quantifying multiple analytes at the point-of-care using a magneto-immunoassay in lateral flow format. PMID:25587419

  1. Electrochemical lateral flow immunosensor for detection and quantification of dengue NS1 protein.

    PubMed

    Sinawang, Prima Dewi; Rai, Varun; Ionescu, Rodica E; Marks, Robert S

    2016-03-15

    An Electrochemical Lateral Flow Immunosensor (ELFI) is developed combining screen-printed gold electrodes (SPGE) enabling quantification together with the convenience of a lateral flow test strip. A cellulose glassy fiber paper conjugate pad retains the marker immunoelectroactive nanobeads which will bind to the target analyte of interest. The specific immunorecognition event continues to occur along the lateral flow bed until reaching the SPGE-capture antibodies at the end of the cellulosic lateral flow strip. The rationale of the immunoassay consists in the analyte antigen NS1 protein being captured selectively and specifically by the dengue NS1 antibody conjugated onto the immunonanobeads thus forming an immunocomplex. With the aid of a running buffer, the immunocomplexes flow and reach the immuno-conjugated electrode surface and form specific sandwich-type detection due to specific, molecular recognition, while unbound beads move along past the electrodes. The successful sandwich immunocomplex formation is then recorded electrochemically. Specific detection of NS1 is translated into an electrochemical signal contributed by a redox label present on the bead-immobilized detection dengue NS1 antibody while a proportional increase of faradic current is observed with increase in analyte NS1 protein concentration. The first generation ELFI prototype is simply assembled in a cassette and successfully demonstrates wide linear range over a concentration range of 1-25 ng/mL with an ultrasensitive detection limit of 0.5 ng/mL for the qualitative and quantitative detection of analyte dengue NS1 protein. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Heat Transfer to Anode of Arc as Function of Transverse Magnetic Field and Lateral Gas Flow Velocity

    NASA Astrophysics Data System (ADS)

    Zama, Yoshiyuki; Shiino, Toru; Ishii, Yoko; Maeda, Yoshifumi; Yamamoto, Shinji; Iwao, Toru

    2016-10-01

    Gas tungsten arc welding has useful joining technology because of high-energy and high-current characteristics. It can be flexible from the transverse magnetic field and lateral gas flow velocity. In this case, the weld defect occurs. In this research, the heat transfer to the anode of the arc as a function of the transverse magnetic field and lateral gas flow velocity is elucidated. That magnetic flux density and lateral gas velocity were varied from 0 to 3 mT and 0 to 50?m?s -1, respectively. The axial plasma gas argon flow rates were 3?slm. A transverse magnetic field is applied to the arc using Helmholtz coil. The anode is used by a water-cooled copper plate, and the heat transfer is measured by temperature of cooled water. As a result, the arc is deflected by the Lorentz force and lateral gas convection. Thus, the heat transfer to the anode of the arc decreases with increasing the transverse magnetic field and lateral gas flow velocity. In addition, the heat transfer to the anode changes with different attachments modes. The lateral gas flow causes a convective heat loss from the arc to the chamber walls.

  3. Quantum-Dot-Based Lateral Flow Immunoassay for Detection of Neonicotinoid Residues in Tea Leaves.

    PubMed

    Wang, Shuangjie; Liu, Ying; Jiao, Shasha; Zhao, Ying; Guo, Yirong; Wang, Mengcen; Zhu, Guonian

    2017-11-22

    Neonicotinoid insecticides are commonly used for pest control on tea plantations as a result of their broad-spectrum activity. However, neonicotinoid residues released from tea leaves into tea infusions pose a dietary risk to consumers. Therefore, a rapid, sensitive, and reliable on-site detection method for neonicotinoids is needed. We developed a quantum-dot-based fluorescent lateral flow immunochromatographic strip (LFICS) combined with a broad-specific antibody for detection of typical neonicotinoids (imidacloprid, imidaclothiz, and clothianidin), with sensitivities [50% inhibitory concentration (IC 50 )] of 0.104-0.33 ng/mL and visual detection limits of 0.5-1 ng/mL. The strip assay could be completed in less than 30 min. Using the LFICS to analyze spiked tea samples (green tea, black tea, and oolong tea), the average recovery of the three neonicotinoids ranged between 71 and 111%, with the coefficient of variation below 12%. The results from the LFICS tests for field samples were consistent with results from ultraperformance liquid chromatography-tandem mass spectrometry. The newly developed strip is a useful tool for the on-site detection of neonicotinoid residues in tea.

  4. Man-made flows from a fish's perspective: autonomous classification of turbulent fishway flows with field data collected using an artificial lateral line.

    PubMed

    Tuhtan, Jeffrey A; Fuentes-Perez, Juan Francisco; Toming, Gert; Schneider, Matthias; Schwarzenberger, Richard; Schletterer, Martin; Kruusmaa, Maarja

    2018-05-25

    The lateral line system provides fish with advanced mechanoreception over a wide range of flow conditions. Inspired by the abilities of their biological counterparts, artificial lateral lines have been developed and tested exclusively under laboratory settings. Motivated by the lack of flow measurements taken in the field which consider fluid-body interactions, we built a fish-shaped lateral line probe. The device is outfitted with 11 high-speed (2.5 kHz) time-synchronized pressure transducers, and designed to capture and classify flows in fish passage structures. A total of 252 field measurements, each with a sample size of 132 000 discrete sensor readings were recorded in the slots and across the pools of vertical slot fishways. These data were used to estimate the time-averaged flow velocity (R 2   =  0.952), which represents the most common metric to assess fishway flows. The significant contribution of this work is the creation and application of hydrodynamic signatures generated by the spatial distribution of pressure fluctuations on the fish-shaped body. The signatures are based on the collection of the pressure fluctuations' probability distributions, and it is shown that they can be used to automatically classify distinct flow regions within the pools of three different vertical slot fishways. For the first time, field data from operational fishway measurements are sampled and classified using an artificial lateral line, providing a completely new source of bioinspired flow information.

  5. Optimized Lateral Flow Immunoassay Reader for the Detection of Infectious Diseases in Developing Countries.

    PubMed

    Pilavaki, Evdokia; Demosthenous, Andreas

    2017-11-20

    Detection and control of infectious diseases is a major problem, especially in developing countries. Lateral flow immunoassays can be used with great success for the detection of infectious diseases. However, for the quantification of their results an electronic reader is required. This paper presents an optimized handheld electronic reader for developing countries. It features a potentially low-cost, low-power, battery-operated device with no added optical accessories. The operation of this proof of concept device is based on measuring the reflected light from the lateral flow immunoassay and translating it into the concentration of the specific analyte of interest. Characterization of the surface of the lateral flow immunoassay has been performed in order to accurately model its response to the incident light. Ray trace simulations have been performed to optimize the system and achieve maximum sensitivity by placing all the components in optimum positions. A microcontroller enables all the signal processing to be performed on the device and a Bluetooth module allows transmission of the results wirelessly to a mobile phone app. Its performance has been validated using lateral flow immunoassays with influenza A nucleoprotein in the concentration range of 0.5 ng/mL to 200 ng/mL.

  6. Mitigating the Hook Effect in Lateral Flow Sandwich Immunoassays Using Real-Time Reaction Kinetics.

    PubMed

    Rey, Elizabeth G; O'Dell, Dakota; Mehta, Saurabh; Erickson, David

    2017-05-02

    The quantification of analyte concentrations using lateral flow assays is a low-cost and user-friendly alternative to traditional lab-based assays. However, sandwich-type immunoassays are often limited by the high-dose hook effect, which causes falsely low results when analytes are present at very high concentrations. In this paper, we present a reaction kinetics-based technique that solves this problem, significantly increasing the dynamic range of these devices. With the use of a traditional sandwich lateral flow immunoassay, a portable imaging device, and a mobile interface, we demonstrate the technique by quantifying C-reactive protein concentrations in human serum over a large portion of the physiological range. The technique could be applied to any hook effect-limited sandwich lateral flow assay and has a high level of accuracy even in the hook effect range.

  7. Channeling at the base of the lithosphere during the lateral flow of plume material beneath flow line hot spots

    NASA Astrophysics Data System (ADS)

    Sleep, Norman H.

    2008-08-01

    Chains of volcanic edifices lie along flow lines between plume-fed hot spots and the thin lithosphere at ridge axes. Discovery and Euterpe/Musicians Seamounts are two examples. An attractive hypothesis is that buoyant plume material flows along the base of the lithosphere perpendicular to isochrons. The plume material may conceivably flow in a broad front or flow within channels convectively eroded into the base to the lithosphere. A necessary but not sufficient condition for convective channeling is that the expected stagnant-lid heat flow for the maximum temperature of the plume material is comparable to the half-space surface heat flow of the oceanic lithosphere. Two-dimensional and three-dimensional numerical calculations confirm this inference. A second criterion for significant convective erosion is that it needs to occur before the plume material thins by lateral spreading. Scaling relationships indicate spreading and convection are closely related. Mathematically, the Nusselt number (ratio of convective to conductive heat flow in the plume material) scales with the flux (volume per time per length of flow front) of the plume material. A blob of unconfined plume material thus spreads before the lithosphere thins much and evolves to a slowly spreading and slowly convecting warm region in equilibrium with conduction into the base of the overlying lithosphere. Three-dimensional calculations illustrate this long-lasting (and hence observable) state of plume material away from its plume source. A different flow domain occurs around a stationary hot plume that continuously supplies hot material. The plume convectively erodes the overlying lithosphere, trapping the plume material near its orifice. The region of lithosphere underlain by plume material grows toward the ridge axis and laterally by convective thinning of the lithosphere at its edges. The hottest plume material channels along flow lines. Geologically, the regions of lithosphere underlain by either warm

  8. Magnetic Control of Lateral Migration of Ellipsoidal Microparticles in Microscale Flows

    NASA Astrophysics Data System (ADS)

    Zhou, Ran; Sobecki, Christopher A.; Zhang, Jie; Zhang, Yanzhi; Wang, Cheng

    2017-08-01

    Precise manipulations of nonspherical microparticles by shape have diverse applications in biology and biomedical engineering. Here, we study lateral migration of ellipsoidal paramagnetic microparticles in low-Reynolds-number flows under uniform magnetic fields. We show that magnetically induced torque alters the rotation dynamics of the particle and results in shape-dependent lateral migration. By adjusting the direction of the magnetic field, we demonstrate versatile control of the symmetric and asymmetric rotation of the particles, thereby controlling the direction of the particle's lateral migration. The particle rotations are experimentally measured, and their symmetry or asymmetry characteristics agree well with the prediction from a simple theory. The lateral migration mechanism is found to be valid for nonmagnetic particles suspended in a ferrofluid. Finally, we demonstrate shape-based sorting of microparticles by exploiting the proposed migration mechanism.

  9. Fluorescent carbon nanoparticle-based lateral flow biosensor for ultrasensitive detection of DNA.

    PubMed

    Takalkar, Sunitha; Baryeh, Kwaku; Liu, Guodong

    2017-12-15

    We report a fluorescent carbon nanoparticle (FCN)-based lateral flow biosensor for ultrasensitive detection of DNA. Fluorescent carbon nanoparticle with a diameter of around 15nm was used as a tag to label a detection DNA probe, which was complementary with the part of target DNA. A capture DNA probe was immobilized on the test zone of the lateral flow biosensor. Sandwich-type hybridization reactions among the FCN-labeled DNA probe, target DNA and capture DNA probe were performed on the lateral flow biosensor. In the presence of target DNA, FCNs were captured on the test zone of the biosensor and the fluorescent intensity of the captured FCNs was measured with a portable fluorescent reader. After systematic optimizations of experimental parameters (the components of running buffers, the concentration of detection DNA probe used in the preparation of FCN-DNA conjugates, the amount of FCN-DNA dispensed on the conjugate pad and the dispensing cycles of the capture DNA probes on the test-zone), the biosensor could detect a minimum concentration of 0.4 fM DNA. This study provides a rapid and low-cost approach for DNA detection with high sensitivity, showing great promise for clinical application and biomedical diagnosis. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. In vivo lateral blood flow velocity measurement using speckle size estimation.

    PubMed

    Xu, Tiantian; Hozan, Mohsen; Bashford, Gregory R

    2014-05-01

    In previous studies, we proposed blood measurement using speckle size estimation, which estimates the lateral component of blood flow within a single image frame based on the observation that the speckle pattern corresponding to blood reflectors (typically red blood cells) stretches (i.e., is "smeared") if blood flow is in the same direction as the electronically controlled transducer line selection in a 2-D image. In this observational study, the clinical viability of ultrasound blood flow velocity measurement using speckle size estimation was investigated and compared with that of conventional spectral Doppler of carotid artery blood flow data collected from human patients in vivo. Ten patients (six male, four female) were recruited. Right carotid artery blood flow data were collected in an interleaved fashion (alternating Doppler and B-mode A-lines) with an Antares Ultrasound Imaging System and transferred to a PC via the Axius Ultrasound Research Interface. The scanning velocity was 77 cm/s, and a 4-s interval of flow data were collected from each subject to cover three to five complete cardiac cycles. Conventional spectral Doppler data were collected simultaneously to compare with estimates made by speckle size estimation. The results indicate that the peak systolic velocities measured with the two methods are comparable (within ±10%) if the scan velocity is greater than or equal to the flow velocity. When scan velocity is slower than peak systolic velocity, the speckle stretch method asymptotes to the scan velocity. Thus, the speckle stretch method is able to accurately measure pure lateral flow, which conventional Doppler cannot do. In addition, an initial comparison of the speckle size estimation and color Doppler methods with respect to computational complexity and data acquisition time indicated potential time savings in blood flow velocity estimation using speckle size estimation. Further studies are needed for calculation of the speckle stretch method

  11. An evaluation of the hydrologic relevance of lateral flow in snow at hillslope and catchment scales

    Treesearch

    David Eiriksson; Michael Whitson; Charles H. Luce; Hans Peter Marshall; John Bradford; Shawn G. Benner; Thomas Black; Hank Hetrick; James P. McNamara

    2013-01-01

    Lateral downslope flow in snow during snowmelt and rain-on-snow (ROS) events is a well-known phenomenon, yet its relevance to water redistribution at hillslope and catchment scales is not well understood. We used dye tracers, geophysical methods, and hydrometric measurements to describe the snow properties that promote lateral flow, assess the relative velocities of...

  12. Experimental investigation of lateral forces induced by flow through model labyrinth glands

    NASA Technical Reports Server (NTRS)

    Leong, Y. M. M. S.; Brown, R. D.

    1984-01-01

    The lateral forces induced by flow through model labyrinth glands were investigated. Circumferential pressure distributions, lateral forces and stiffness coefficients data obtained are discussed. The force system is represented as a negative spring and a tangential force orthogonal to eccentricity. The magnitude of these forces are dependent on eccentricity, entry swirl, rotor peripheral velocity and seal size. A pressure equalization chamber at midgland tests should in significantly reduced forces and stiffness coefficients.

  13. Image-based modelling of lateral magma flow: the Basement Sill, Antarctica

    PubMed Central

    Mirhadizadeh, Seyed

    2017-01-01

    The McMurdo Dry Valleys magmatic system, Antarctica, provides a world-class example of pervasive lateral magma flow on a continental scale. The lowermost intrusion (Basement Sill) offers detailed sections through the now frozen particle microstructure of a congested magma slurry. We simulated the flow regime in two and three dimensions using numerical models built on a finite-element mesh derived from field data. The model captures the flow behaviour of the Basement Sill magma over a viscosity range of 1–104 Pa s where the higher end (greater than or equal to 102 Pa s) corresponds to a magmatic slurry with crystal fractions varying between 30 and 70%. A novel feature of the model is the discovery of transient, low viscosity (less than or equal to 50 Pa s) high Reynolds number eddies formed along undulating contacts at the floor and roof of the intrusion. Numerical tracing of particle orbits implies crystals trapped in eddies segregate according to their mass density. Recovered shear strain rates (10−3–10−5 s−1) at viscosities equating to high particle concentrations (around more than 40%) in the Sill interior point to shear-thinning as an explanation for some types of magmatic layering there. Model transport rates for the Sill magmas imply a maximum emplacement time of ca 105 years, consistent with geochemical evidence for long-range lateral flow. It is a theoretically possibility that fast-flowing magma on a continental scale will be susceptible to planetary-scale rotational forces. PMID:28573002

  14. Image-based modelling of lateral magma flow: the Basement Sill, Antarctica.

    PubMed

    Petford, Nick; Mirhadizadeh, Seyed

    2017-05-01

    The McMurdo Dry Valleys magmatic system, Antarctica, provides a world-class example of pervasive lateral magma flow on a continental scale. The lowermost intrusion (Basement Sill) offers detailed sections through the now frozen particle microstructure of a congested magma slurry. We simulated the flow regime in two and three dimensions using numerical models built on a finite-element mesh derived from field data. The model captures the flow behaviour of the Basement Sill magma over a viscosity range of 1-10 4  Pa s where the higher end (greater than or equal to 10 2  Pa s) corresponds to a magmatic slurry with crystal fractions varying between 30 and 70%. A novel feature of the model is the discovery of transient, low viscosity (less than or equal to 50 Pa s) high Reynolds number eddies formed along undulating contacts at the floor and roof of the intrusion. Numerical tracing of particle orbits implies crystals trapped in eddies segregate according to their mass density. Recovered shear strain rates (10 -3 -10 -5  s -1 ) at viscosities equating to high particle concentrations (around more than 40%) in the Sill interior point to shear-thinning as an explanation for some types of magmatic layering there. Model transport rates for the Sill magmas imply a maximum emplacement time of ca 10 5 years, consistent with geochemical evidence for long-range lateral flow. It is a theoretically possibility that fast-flowing magma on a continental scale will be susceptible to planetary-scale rotational forces.

  15. Distributed flow estimation and closed-loop control of an underwater vehicle with a multi-modal artificial lateral line.

    PubMed

    DeVries, Levi; Lagor, Francis D; Lei, Hong; Tan, Xiaobo; Paley, Derek A

    2015-03-25

    Bio-inspired sensing modalities enhance the ability of autonomous vehicles to characterize and respond to their environment. This paper concerns the lateral line of cartilaginous and bony fish, which is sensitive to fluid motion and allows fish to sense oncoming flow and the presence of walls or obstacles. The lateral line consists of two types of sensing modalities: canal neuromasts measure approximate pressure gradients, whereas superficial neuromasts measure local flow velocities. By employing an artificial lateral line, the performance of underwater sensing and navigation strategies is improved in dark, cluttered, or murky environments where traditional sensing modalities may be hindered. This paper presents estimation and control strategies enabling an airfoil-shaped unmanned underwater vehicle to assimilate measurements from a bio-inspired, multi-modal artificial lateral line and estimate flow properties for feedback control. We utilize potential flow theory to model the fluid flow past a foil in a uniform flow and in the presence of an upstream obstacle. We derive theoretically justified nonlinear estimation strategies to estimate the free stream flowspeed, angle of attack, and the relative position of an upstream obstacle. The feedback control strategy uses the estimated flow properties to execute bio-inspired behaviors including rheotaxis (the tendency of fish to orient upstream) and station-holding (the tendency of fish to position behind an upstream obstacle). A robotic prototype outfitted with a multi-modal artificial lateral line composed of ionic polymer metal composite and embedded pressure sensors experimentally demonstrates the distributed flow sensing and closed-loop control strategies.

  16. Lateral flow immunoassay for the rapid detection of citrus tristeza virus

    USDA-ARS?s Scientific Manuscript database

    A lateral flow methodology was developed using gold nanoparticles for rapid detection of Citrus tristeza virus (CTV). The test strip was based on a sandwich immunoassay and could be accomplished within 10 minutes. A sample was considered negative for CTV when only the control line appeared; whereas,...

  17. A flow-through column electrolytic cell for supercritical fluid chromatography.

    PubMed

    Yamamoto, Kazuhiro; Ueki, Tatsuya; Higuchi, Naoyuki; Takahashi, Kouji; Kotani, Akira; Hakamata, Hideki

    2017-10-01

    A novel flow-through column electrolytic cell was proposed as a detector to obtain current signals for supercritical fluid chromatography. The electrochemical cell consisted of two electrodes and its holder, and a working and a counter electrode were fabricated from 192 carbon strings, which were composed of 400 carbon fibers of 10 μm in diameter filled into a heat-shrinkable tube. These electrodes were placed in the center of a holder made from polyether ether ketone blocks and they were separated by polytetrafluoroethylene membrane filters. To evaluate the sensitivity of this cell, a standard solution of ferrocene was injected into the supercritical fluid chromatography system connected to the electrolytic cell. The ferrocene was eluted through a silica gel column using a mixture of a mobile phase of supercritical CO 2 and a modifier of methanol containing ammonium acetate. The current peak area of ferrocene correlated to the ferrocene concentration in the range of 10-400 μmol/L (r = 0.999). Moreover, the limit of detection on the column estimated from a signal-to-noise ratio of 3 was 9.8  × 10 -13  mol. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Porcine Circovirus (PCV) Removal by Q Sepharose Fast Flow Chromatography

    PubMed Central

    Yang, Bin; Wang, Hua; Ho, Cintia; Lester, Philip; Chen, Qi; Neske, Florian; Baylis, Sally A; Blümel, Johannes

    2013-01-01

    The recently discovered contamination of oral rotavirus vaccines led to exposure of millions of infants to porcine circovirus (PCV). PCV was not detected by conventional virus screening tests. Regulatory agencies expect exclusion of adventitious viruses from biological products. Therefore, methods for inactivation/removal of viruses have to be implemented as an additional safety barrier whenever feasible. However, inactivation or removal of PCV is difficult. PCV is highly resistant to widely used physicochemical inactivation procedures. Circoviruses such as PCV are the smallest viruses known and are not expected to be effectively removed by currently-used virus filters due to the small size of the circovirus particles. Anion exchange chromatography such as Q Sepharose® Fast Flow (QSFF) has been shown to effectively remove a range of viruses including parvoviruses. In this study, we investigated PCV1 removal by virus filtration and by QSFF chromatography. As expected, PCV1 could not be effectively removed by virus filtration. However, PCV1 could be effectively removed by QSFF as used during the purification of monoclonal antibodies (mAbs) and a log10 reduction value (LRV) of 4.12 was obtained. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1464–1471, 2013 PMID:24039195

  19. High-Speed Lateral Flow Strategy for a Fast Biosensing with an Improved Selectivity and Binding Affinity.

    PubMed

    Cho, Dong Guk; Yoo, Haneul; Lee, Haein; Choi, Yeol Kyo; Lee, Minju; Ahn, Dong June; Hong, Seunghun

    2018-05-10

    We report a high-speed lateral flow strategy for a fast biosensing with an improved selectivity and binding affinity even under harsh conditions. In this strategy, biosensors were fixed at a location away from the center of a round shape disk, and the disk was rotated to create the lateral flow of a target solution on the biosensors during the sensing measurements. Experimental results using the strategy showed high reaction speeds, high binding affinity, and low nonspecific adsorptions of target molecules to biosensors. Furthermore, binding affinity between target molecules and sensing molecules was enhanced even in harsh conditions such as low pH and low ionic strength conditions. These results show that the strategy can improve the performance of conventional biosensors by generating high-speed lateral flows on a biosensor surface. Therefore, our strategy can be utilized as a simple but powerful tool for versatile bio and medical applications.

  20. An optofluidic metasurface for lateral flow-through detection of breast cancer biomarker.

    PubMed

    Wang, Yifei; Ali, Md Azahar; Chow, Edmond K C; Dong, Liang; Lu, Meng

    2018-06-01

    The rapid growth of point-of-care tests demands for biosensors with high sensitivity and small size. This paper demonstrates an optofluidic metasurface that combines silicon-on-insulator (SOI) nanophotonics and nanofluidics to realize a high-performance, lateral flow-through biosensor. The metasurface is made of a periodic array of silicon nanoposts on an SOI substrate, and functionalized with specific receptor molecules. Bonding of a polydimethylsiloxane slab directly onto the surface results in an ultracompact biosensor, where analyte solutions are restricted to flow only in the space between the nanoposts. No flow exists above the nanoposts. This sensor design overcomes the issue with diffusion-limited detection of many other biosensors. The lateral flow-through feature, in conjunction with high-Q resonance modes associated with optical bound states of the metasurface, offers an improved sensitivity to subtle molecule-bonding induced changes in refractive index. The device exhibits a resonance mode around 1550 nm wavelength and provides an index sensitivity of 720 nm/RIU. Biosensing is conducted to detect the epidermal growth factor receptor 2 (ErbB2), a protein biomarker for early-stage breast cancer screening, by monitoring resonance wavelength shifts in response to specific analyte-ligand binding events at the metasurface. The limit of detection of the device is 0.7 ng mL -1 for ErbB2. Copyright © 2018 Elsevier B.V. All rights reserved.

  1. Automated Low-Cost Smartphone-Based Lateral Flow Saliva Test Reader for Drugs-of-Abuse Detection

    PubMed Central

    Carrio, Adrian; Sampedro, Carlos; Sanchez-Lopez, Jose Luis; Pimienta, Miguel; Campoy, Pascual

    2015-01-01

    Lateral flow assay tests are nowadays becoming powerful, low-cost diagnostic tools. Obtaining a result is usually subject to visual interpretation of colored areas on the test by a human operator, introducing subjectivity and the possibility of errors in the extraction of the results. While automated test readers providing a result-consistent solution are widely available, they usually lack portability. In this paper, we present a smartphone-based automated reader for drug-of-abuse lateral flow assay tests, consisting of an inexpensive light box and a smartphone device. Test images captured with the smartphone camera are processed in the device using computer vision and machine learning techniques to perform automatic extraction of the results. A deep validation of the system has been carried out showing the high accuracy of the system. The proposed approach, applicable to any line-based or color-based lateral flow test in the market, effectively reduces the manufacturing costs of the reader and makes it portable and massively available while providing accurate, reliable results. PMID:26610513

  2. Automated Low-Cost Smartphone-Based Lateral Flow Saliva Test Reader for Drugs-of-Abuse Detection.

    PubMed

    Carrio, Adrian; Sampedro, Carlos; Sanchez-Lopez, Jose Luis; Pimienta, Miguel; Campoy, Pascual

    2015-11-24

    Lateral flow assay tests are nowadays becoming powerful, low-cost diagnostic tools. Obtaining a result is usually subject to visual interpretation of colored areas on the test by a human operator, introducing subjectivity and the possibility of errors in the extraction of the results. While automated test readers providing a result-consistent solution are widely available, they usually lack portability. In this paper, we present a smartphone-based automated reader for drug-of-abuse lateral flow assay tests, consisting of an inexpensive light box and a smartphone device. Test images captured with the smartphone camera are processed in the device using computer vision and machine learning techniques to perform automatic extraction of the results. A deep validation of the system has been carried out showing the high accuracy of the system. The proposed approach, applicable to any line-based or color-based lateral flow test in the market, effectively reduces the manufacturing costs of the reader and makes it portable and massively available while providing accurate, reliable results.

  3. A bio-inspired real-time capable artificial lateral line system for freestream flow measurements.

    PubMed

    Abels, C; Qualtieri, A; De Vittorio, M; Megill, W M; Rizzi, F

    2016-06-03

    To enhance today's artificial flow sensing capabilities in aerial and underwater robotics, future robots could be equipped with a large number of miniaturized sensors distributed over the surface to provide high resolution measurement of the surrounding fluid flow. In this work we show a linear array of closely separated bio-inspired micro-electro-mechanical flow sensors whose sensing mechanism is based on a piezoresistive strain-gauge along a stress-driven cantilever beam, mimicking the biological superficial neuromasts found in the lateral line organ of fishes. Aiming to improve state-of-the-art flow sensing capability in autonomously flying and swimming robots, our artificial lateral line system was designed and developed to feature multi-parameter freestream flow measurements which provide information about (1) local flow velocities as measured by the signal amplitudes from the individual cantilevers as well as (2) propagation velocity, (3) linear forward/backward direction along the cantilever beam orientation and (4) periodicity of pulses or pulse trains determined by cross-correlating sensor signals. A real-time capable cross-correlation procedure was developed which makes it possible to extract freestream flow direction and velocity information from flow fluctuations. The computed flow velocities deviate from a commercial system by 0.09 m s(-1) at 0.5 m s(-1) and 0.15 m s(-1) at 1.0 m s(-1) flow velocity for a sampling rate of 240 Hz and a sensor distance of 38 mm. Although experiments were performed in air, the presented flow sensing system can be applied to underwater vehicles as well, once the sensors are embedded in a waterproof micro-electro-mechanical systems package.

  4. A Microwave Flow Detector for Gradient Elution Liquid Chromatography.

    PubMed

    Ye, Duye; Wang, Weizheng; Moline, David; Islam, Md Saiful; Chen, Feng; Wang, Pingshan

    2017-10-17

    This study presents a microwave flow detector technique for liquid chromatography (LC) application. The detector is based on a tunable microwave interferometer (MIM) with a vector network analyzer (VNA) for signal measurement and a computer for system control. A microstrip-line-based 0.3 μL flow cell is built and incorporated into the MIM. With syringe pump injection, the detector is evaluated by measuring a few common chemicals in DI water at multiple frequencies from 0.98 to 7.09 GHz. Less than 30 ng minimum detectable quantity (MDQ) is demonstrated. An algorithm is provided and used to obtain sample dielectric permittivity at each frequency point. When connected to a commercial HPLC system and injected with a 10 μL aliquot of 10 000 ppm caffeine DI-water solution, the microwave detector yields a signal-to-noise ratio (SNR) up to 10 under isocratic and gradient elution operations. The maximum sampling rate is 20 Hz. The measurements show that MIM tuning, aided by a digital tunable attenuator (DTA), can automatically adjust MIM operation to retain detector sensitivity when mobile phase changes. Furthermore, the detector demonstrates a capability to quantify coeluted vitamin E succinate (VES) and vitamin D 3 (VD 3 ).

  5. Assessing lateral flows and solute transport during floods in a conduit-flow-dominated karst system using the inverse problem for the advection-diffusion equation

    NASA Astrophysics Data System (ADS)

    Cholet, Cybèle; Charlier, Jean-Baptiste; Moussa, Roger; Steinmann, Marc; Denimal, Sophie

    2017-07-01

    The aim of this study is to present a framework that provides new ways to characterize the spatio-temporal variability of lateral exchanges for water flow and solute transport in a karst conduit network during flood events, treating both the diffusive wave equation and the advection-diffusion equation with the same mathematical approach, assuming uniform lateral flow and solute transport. A solution to the inverse problem for the advection-diffusion equations is then applied to data from two successive gauging stations to simulate flows and solute exchange dynamics after recharge. The study site is the karst conduit network of the Fourbanne aquifer in the French Jura Mountains, which includes two reaches characterizing the network from sinkhole to cave stream to the spring. The model is applied, after separation of the base from the flood components, on discharge and total dissolved solids (TDSs) in order to assess lateral flows and solute concentrations and compare them to help identify water origin. The results showed various lateral contributions in space - between the two reaches located in the unsaturated zone (R1), and in the zone that is both unsaturated and saturated (R2) - as well as in time, according to hydrological conditions. Globally, the two reaches show a distinct response to flood routing, with important lateral inflows on R1 and large outflows on R2. By combining these results with solute exchanges and the analysis of flood routing parameters distribution, we showed that lateral inflows on R1 are the addition of diffuse infiltration (observed whatever the hydrological conditions) and localized infiltration in the secondary conduit network (tributaries) in the unsaturated zone, except in extreme dry periods. On R2, despite inflows on the base component, lateral outflows are observed during floods. This pattern was attributed to the concept of reversal flows of conduit-matrix exchanges, inducing a complex water mixing effect in the saturated zone

  6. Sensitive Detection of Norovirus Using Phage Nanoparticle Reporters in Lateral-Flow Assay

    PubMed Central

    Hagström, Anna E. V.; Garvey, Gavin; Paterson, Andrew S.; Dhamane, Sagar; Adhikari, Meena; Estes, Mary K.; Strych, Ulrich; Kourentzi, Katerina; Atmar, Robert L.; Willson, Richard C.

    2015-01-01

    Noroviruses are recognized worldwide as the principal cause of acute, non-bacterial gastroenteritis, resulting in 19-21 million cases of disease every year in the United States. Noroviruses have a very low infectious dose, a short incubation period, high resistance to traditional disinfection techniques and multiple modes of transmission, making early, point-of-care detection essential for controlling the spread of the disease. The traditional diagnostic tools, electron microscopy, RT-PCR and ELISA require sophisticated and expensive instrumentation, and are considered too laborious and slow to be useful during severe outbreaks. In this paper we describe the development of a new, rapid and sensitive lateral-flow assay using labeled phage particles for the detection of the prototypical norovirus GI.1 (Norwalk), with a limit of detection of 107 virus-like particles per mL, one hundred-fold lower than a conventional gold nanoparticle lateral-flow assay using the same antibody pair. PMID:25978622

  7. Comparison of standard- and nano-flow liquid chromatography platforms for MRM-based quantitation of putative plasma biomarker proteins.

    PubMed

    Percy, Andrew J; Chambers, Andrew G; Yang, Juncong; Domanski, Dominik; Borchers, Christoph H

    2012-09-01

    The analytical performance of a standard-flow ultra-high-performance liquid chromatography (UHPLC) and a nano-flow high-performance liquid chromatography (HPLC) system, interfaced to the same state-of-the-art triple-quadrupole mass spectrometer, were compared for the multiple reaction monitoring (MRM)-mass spectrometry (MS)-based quantitation of a panel of 48 high-to-moderate-abundance cardiovascular disease-related plasma proteins. After optimization of the MRM transitions for sensitivity and testing for chemical interference, the optimum sensitivity, loading capacity, gradient, and retention-time reproducibilities were determined. We previously demonstrated the increased robustness of the standard-flow platform, but we expected that the standard-flow platform would have an overall lower sensitivity. This study was designed to determine if this decreased sensitivity could be compensated for by increased sample loading. Significantly fewer interferences with the MRM transitions were found for the standard-flow platform than for the nano-flow platform (2 out of 103 transitions compared with 42 out of 103 transitions, respectively), which demonstrates the importance of interference-testing when nano-flow systems are used. Using only interference-free transitions, 36 replicate LC/MRM-MS analyses resulted in equal signal reproducibilities between the two platforms (9.3 % coefficient of variation (CV) for 88 peptide targets), with superior retention-time precision for the standard-flow platform (0.13 vs. 6.1 % CV). Surprisingly, for 41 of the 81 proteotypic peptides in the final assay, the standard-flow platform was more sensitive while for 9 of 81 the nano-flow platform was more sensitive. For these 81 peptides, there was a good correlation between the two sets of results (R(2) = 0.98, slope = 0.97). Overall, the standard-flow platform had superior performance metrics for most peptides, and is a good choice if sufficient sample is available.

  8. How bumblebees use lateral and ventral optic flow cues for position control in environments of different proximity.

    PubMed

    Linander, Nellie; Baird, Emily; Dacke, Marie

    2017-05-01

    Flying insects frequently navigate through environments of different complexity. In this study, buff-tailed bumblebees (Bombus terrestris L.) were trained to fly along tunnels of different widths, from 60 to 240 cm. In tunnel widths of 60 and 120 cm, bumblebees control their lateral position by balancing the magnitude of translational optic flow experienced in the lateral visual field of each eye. In wider tunnels, bumblebees use translational optic flow cues in the ventral visual field to control their lateral position and to steer along straight tracks. Our results also suggest that bumblebees prefer to fly over surfaces that provide strong ventral optic flow cues, rather than over featureless ones. Together, these strategies allow bumblebees to minimize the risk of collision and to maintain relatively straight flight paths in a broad range of environments.

  9. Statistical theory of chromatography: new outlooks for affinity chromatography.

    PubMed Central

    Denizot, F C; Delaage, M A

    1975-01-01

    We have developed further the statistical approach to chromatography initiated by Giddings and Eyring, and applied it to affinity chromatography. By means of a convenient expression of moments the convergence towards the Laplace-Gauss distribution has been established. The Gaussian character is not preserved if other causes of dispersion are taken into account, but expressions of moments can be obtained in a generalized form. A simple procedure is deduced for expressing the fundamental constants of the model in terms of purely experimental quantities. Thus, affinity chromatography can be used to determine rate constants of association and dissociation in a range considered as the domain of the stopped-flow methods. PMID:1061072

  10. Lateral and subsurface flows impact arctic coastal plain lake water budgets

    USGS Publications Warehouse

    Koch, Joshua C.

    2016-01-01

    Arctic thaw lakes are an important source of water for aquatic ecosystems, wildlife, and humans. Many recent studies have observed changes in Arctic surface waters related to climate warming and permafrost thaw; however, explaining the trends and predicting future responses to warming is difficult without a stronger fundamental understanding of Arctic lake water budgets. By measuring and simulating surface and subsurface hydrologic fluxes, this work quantified the water budgets of three lakes with varying levels of seasonal drainage, and tested the hypothesis that lateral and subsurface flows are a major component of the post-snowmelt water budgets. A water budget focused only on post-snowmelt surface water fluxes (stream discharge, precipitation, and evaporation) could not close the budget for two of three lakes, even when uncertainty in input parameters was rigorously considered using a Monte Carlo approach. The water budgets indicated large, positive residuals, consistent with up to 70% of mid-summer inflows entering lakes from lateral fluxes. Lateral inflows and outflows were simulated based on three processes; supra-permafrost subsurface inflows from basin-edge polygonal ground, and exchange between seasonally drained lakes and their drained margins through runoff and evapotranspiration. Measurements and simulations indicate that rapid subsurface flow through highly conductive flowpaths in the polygonal ground can explain the majority of the inflow. Drained lakes were hydrologically connected to marshy areas on the lake margins, receiving water from runoff following precipitation and losing up to 38% of lake efflux to drained margin evapotranspiration. Lateral fluxes can be a major part of Arctic thaw lake water budgets and a major control on summertime lake water levels. Incorporating these dynamics into models will improve our ability to predict lake volume changes, solute fluxes, and habitat availability in the changing Arctic.

  11. Macro-/Nano- Materials Based Ultrasensitive Lateral Flow Nucleic Acid Biosensors

    NASA Astrophysics Data System (ADS)

    Takalkar, Sunitha

    Ultrasensitive detection of nucleic acids plays a very important role in the field of molecular diagnosis for the detection of various diseases. Lateral flow biosensors (LFB) are convenient, easy-to-use, patient friendly forms of detection methods offering rapid and convenient clinical testing in close proximity to the patients thus drawing a lot of attention in different areas of research over the years. In comparison with the traditional immunoassays, the nucleic acid based lateral flow biosensors (NABLFB) has several advantages in terms of stability and interference capabilities. NABLFB utilizes nucleic acid probes as the bio-recognition element. The target analyte typically is the oligonucleotide like the DNA, mRNA, miRNA which are among the nucleic acid secretions by the tumor cells when it comes to detection of cancer. Traditionally gold nanoparticles (GNPs) have been used as labels for conjugating with the detection probes for the qualitative and semi quantitative analysis, the application of GNP-based LFB is limited by its low sensitivity. This dissertation describes the use of different nanomaterials and advanced detection technologies to enhance the sensitivities of the LFB based methods. Silica Nanorods decorated with GNP were synthesized and employed as labels for ultrasensitive detection of miRNA on the LFB. Owing to the biocompatibility and convenience in surface modification of SiNRs, they acted as good carriers to load numerous GNPs. The sensitivity of the GNP-SiNR-based LFSB was enhanced six times compared to the previous GNP-based LFSB. A fluorescent carbon nanoparticle (FCN) was first used as a tag to develop a lateral flow nucleic acid biosensor for ultrasensitive and quantitative detection of nucleic acid samples. Under optimal conditions, the FCN-based LFNAB was capable of detecting minimum 0.4 fM target DNA without complex operations and additional signal amplification. The carbon nanotube was used as a label and carrier of numerous enzyme

  12. Rapid detection of Bacillus anthracis spores using a super-paramagnetic lateral-flow immunological detection system.

    PubMed

    Wang, Dian-Bing; Tian, Bo; Zhang, Zhi-Ping; Deng, Jiao-Yu; Cui, Zong-Qiang; Yang, Rui-Fu; Wang, Xu-Ying; Wei, Hong-Ping; Zhang, Xian-En

    2013-04-15

    There is an urgent need for convenient, sensitive, and specific methods to detect the spores of Bacillus anthracis, the causative agent of anthrax, because of the bioterrorism threat posed by this bacterium. In this study, we firstly develop a super-paramagnetic lateral-flow immunological detection system for B. anthracis spores. This system involves the use of a portable magnetic assay reader, super-paramagnetic iron oxide particles, lateral-flow strips and two different monoclonal antibodies directed against B. anthracis spores. This detection system specifically recognises as few as 400 pure B. anthracis spores in 30 min. This system has a linear range of 4×10³-10⁶ CFU ml⁻¹ and reproducible detection limits of 200 spores mg⁻¹ milk powder and 130 spores mg⁻¹ soil for simulated samples. In addition, this approach shows no obvious cross-reaction with other related Bacillus spores, even at high concentrations, and has no significant dependence on the duration of the storage of the immunological strips. Therefore, this super-paramagnetic lateral-flow immunological detection system is a promising tool for the rapid and sensitive detection of Bacillus anthracis spores under field conditions. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Influence of lateral groundwater flow in a shallow aquifer on eco-hydrological process in a shrub-grass coexistence semiarid area

    NASA Astrophysics Data System (ADS)

    Wang, Siru; Sun, Jinhua; Lei, Huimin; Zhu, Qiande; Jiang, Sanyuan

    2017-04-01

    Topography has a considerable influence on eco-hydrological processes resulting from the patterns of solar radiation distribution and lateral water flow. However, not much quantitative information on the contribution of lateral groundwater flow on ecological processes such as vegetation growth and evapo-transpiration is available. To fill this gap, we used a simple eco-hydrological model based on water balance with a 3D groundwater module that uses Darcy's law. This model was applied to a non-contributing area of 50km2 dominated by grassland and shrubland with an underlying shallow aquifer. It was calibrated using manually and remotely sensed vegetation data and water flux data observed by eddy covariance system of two flux towers as well as water table data obtained from HOBO recorders of 40 wells. The results demonstrate that the maximum hydraulic gradient and the maximum flux of lateral groundwater flow reached to 0.156m m-1 and 0.093m3 s-1 respectively. The average annual maximum LAI in grassland, predominantly in low-lying areas, improved by about 5.9% while that in shrubland, predominantly in high-lying areas, remained the same when lateral groundwater flow is considered adequately compared to the case without considering lateral groundwater flow. They also show that LAI is positively and nonlinearly related to evapotranspiration, and that the greater the magnitude of evapotranspiration, the smaller the rate of increase of LAI. The results suggest that lateral groundwater flow should not be neglected when simulating eco-hydrological process in areas with a shallow aquifer.

  14. Design and application of a fish-shaped lateral line probe for flow measurement

    NASA Astrophysics Data System (ADS)

    Tuhtan, J. A.; Fuentes-Pérez, J. F.; Strokina, N.; Toming, G.; Musall, M.; Noack, M.; Kämäräinen, J. K.; Kruusmaa, M.

    2016-04-01

    We introduce the lateral line probe (LLP) as a measurement device for natural flows. Hydraulic surveys in rivers and hydraulic structures are currently based on time-averaged velocity measurements using propellers or acoustic Doppler devices. The long-term goal is thus to develop a sensor system, which includes spatial gradients of the flow field along a fish-shaped sensor body. Interpreting the biological relevance of a collection of point velocity measurements is complicated by the fact that fish and other aquatic vertebrates experience the flow field through highly dynamic fluid-body interactions. To collect body-centric flow data, a bioinspired fish-shaped probe is equipped with a lateral line pressure sensing array, which can be applied both in the laboratory and in the field. Our objective is to introduce a new type of measurement device for body-centric data and compare its output to estimates of conventional point-based technologies. We first provide the calibration workflow for laboratory investigations. We then provide a review of two velocity estimation workflows, independent of calibration. Such workflows are required as existing field investigations consist of measurements in environments where calibration is not feasible. The mean difference for uncalibrated LLP velocity estimates from 0 to 50 cm/s under in a closed flow tunnel and open channel flume was within 4 cm/s when compared to conventional measurement techniques. Finally, spatial flow maps in a scale vertical slot fishway are compared for the LLP, direct measurements, and 3D numerical models where it was found that the LLP provided a slight overestimation of the current velocity in the jet and underestimated the velocity in the recirculation zone.

  15. Atomic Force Microscope Mediated Chromatography

    NASA Technical Reports Server (NTRS)

    Anderson, Mark S.

    2013-01-01

    The atomic force microscope (AFM) is used to inject a sample, provide shear-driven liquid flow over a functionalized substrate, and detect separated components. This is demonstrated using lipophilic dyes and normal phase chromatography. A significant reduction in both size and separation time scales is achieved with a 25-micron-length column scale, and one-second separation times. The approach has general applications to trace chemical and microfluidic analysis. The AFM is now a common tool for ultra-microscopy and nanotechnology. It has also been demonstrated to provide a number of microfluidic functions necessary for miniaturized chromatography. These include injection of sub-femtoliter samples, fluidic switching, and sheardriven pumping. The AFM probe tip can be used to selectively remove surface layers for subsequent microchemical analysis using infrared and tip-enhanced Raman spectroscopy. With its ability to image individual atoms, the AFM is a remarkably sensitive detector that can be used to detect separated components. These diverse functional components of microfluidic manipulation have been combined in this work to demonstrate AFM mediated chromatography. AFM mediated chromatography uses channel-less, shear-driven pumping. This is demonstrated with a thin, aluminum oxide substrate and a non-polar solvent system to separate a mixture of lipophilic dyes. In conventional chromatographic terms, this is analogous to thin-layer chromatography using normal phase alumina substrate with sheardriven pumping provided by the AFM tip-cantilever mechanism. The AFM detection of separated components is accomplished by exploiting the variation in the localized friction of the separated components. The AFM tip-cantilever provides the mechanism for producing shear-induced flows and rapid pumping. Shear-driven chromatography (SDC) is a relatively new concept that overcomes the speed and miniaturization limitations of conventional liquid chromatography. SDC is based on a

  16. Characterization of branched ultrahigh molar mass polymers by asymmetrical flow field-flow fractionation and size exclusion chromatography.

    PubMed

    Otte, T; Pasch, H; Macko, T; Brüll, R; Stadler, F J; Kaschta, J; Becker, F; Buback, M

    2011-07-08

    The molar mass distribution (MMD) of synthetic polymers is frequently analyzed by size exclusion chromatography (SEC) coupled to multi angle light scattering (MALS) detection. For ultrahigh molar mass (UHM) or branched polymers this method is not sufficient, because shear degradation and abnormal elution effects falsify the calculated molar mass distribution and information on branching. High temperatures above 130 °C have to be applied for dissolution and separation of semi-crystalline materials like polyolefins which requires special hardware setups. Asymmetrical flow field-flow fractionation (AF4) offers the possibility to overcome some of the main problems of SEC due to the absence of an obstructing porous stationary phase. The SEC-separation mainly depends on the pore size distribution of the used column set. The analyte molecules can enter the pores of the stationary phase in dependence on their hydrodynamic volume. The archived separation is a result of the retention time of the analyte species inside SEC-column which depends on the accessibility of the pores, the residence time inside the pores and the diffusion ability of the analyte molecules. The elution order in SEC is typically from low to high hydrodynamic volume. On the contrary AF4 separates according to the diffusion coefficient of the analyte molecules as long as the chosen conditions support the normal FFF-separation mechanism. The separation takes place in an empty channel and is caused by a cross-flow field perpendicular to the solvent flow. The analyte molecules will arrange in different channel heights depending on the diffusion coefficients. The parabolic-shaped flow profile inside the channel leads to different elution velocities. The species with low hydrodynamic volume will elute first while the species with high hydrodynamic volume elute later. The AF4 can be performed at ambient or high temperature (AT-/HT-AF4). We have analyzed one low molar mass polyethylene sample and a number of

  17. Simultaneous determination of aflatoxin B₁, B₂, G₁, and G₂ in corn powder, edible oil, peanut butter, and soy sauce by liquid chromatography with tandem mass spectrometry utilizing turbulent flow chromatography.

    PubMed

    Fan, Sufang; Li, Qiang; Zhang, Xiaoguang; Cui, Xiaobin; Zhang, Dongsheng; Zhang, Yan

    2015-05-01

    A novel fully automated method based on dual column switching using turbulent flow chromatography followed by liquid chromatography with tandem mass spectrometry was developed for the determination of aflatoxin B1 , B2 , G1 , and G2 in corn powder, edible oil, peanut butter, and soy sauce samples. After ultrasound-assisted extraction, samples were directly injected to the chromatographic system and the analytes were concentrated into the clean-up loading column. Through purge switching, the analytes were transferred to the analytical column for subsequent detection by mass spectrometry. Different types of TurboFlow(TM) columns, transfer flow rate, transfer time were optimized. The limits of detection and quantification of this method ranged between 0.2-2.0 and 0.5-4.0 μg/kg for aflatoxins in different matrixes, respectively. Recoveries of aflatoxins were in range of 83-108.1% for all samples, matrix effects were in range of 34.1-104.7%. The developed method has been successfully applied in the analysis of aflatoxin B1 , B2 , G1 , and G2 in real samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Lateral Migration and Rotational Motion of Elliptic Particles in Planar Poiseuille Flow

    NASA Technical Reports Server (NTRS)

    Qi, Dewei; Luo, Li-Shi; Aravamuthan, Raja; Strieder, William; Bushnell, Dennis M. (Technical Monitor)

    2002-01-01

    Simulations of elliptic particulate suspensions in the planar Poiseuille flow are performed by using the lattice Boltzmann equation. Effects of the multi-particle on the lateral migration and rotational motion of both neutrally and non-neutrally buoyant elliptic particles are investigated. Low and intermediate total particle volume fraction f(sub a) = 13%, 15%, and 40% are considered in this work.

  19. An Ultrasensitive Gold Nanoparticle-based Lateral Flow Test for the Detection of Active Botulinum Neurotoxin Type A

    NASA Astrophysics Data System (ADS)

    Liu, Jing; Gao, Shan; Kang, Lin; Ji, Bin; Xin, Wenwen; Kang, Jingjing; Li, Ping; Gao, Jie; Wang, Hanbin; Wang, Jinglin; Yang, Hao

    2017-03-01

    Botulism is a severe and potentially lethal paralytic disease caused by several botulinum neurotoxin-producing Clostridia spp. In China, the majority of the cases caused by botulism were from less-developed rural areas. Here, we designed specific substrate peptides and reconfigured gold nanoparticle-based lateral flow test strip (LFTS) to develop an endopeptidase-based lateral flow assay for the diagnosis of botulism. We performed this lateral flow assay on botulinum neurotoxin-spiked human serum samples. The as-prepared LFTS had excellent performance in the detection of botulinum neurotoxin using only 1 μL of simulated serum, and its sensitivity and specificity were comparable to that of mouse lethality assay. Moreover, the assay takes only half a day and does not require highly trained laboratory staff, specialized facility, or equipment. Finally, our LFTS can be potentially extended to other serotypes of BoNTs by designing specific substrate peptides against the different types of BoNTs. Overall, we demonstrate a strategy by which LFTS and endopeptidase activity assays can be integrated to achieve facile and economic diagnosis of botulism in resource-limited settings.

  20. Detection of Bacillus anthracis spores by super-paramagnetic lateral-flow immunoassays based on "Road Closure".

    PubMed

    Wang, Dian-Bing; Tian, Bo; Zhang, Zhi-Ping; Wang, Xu-Ying; Fleming, Joy; Bi, Li-Jun; Yang, Rui-Fu; Zhang, Xian-En

    2015-05-15

    Detection of Bacillus anthracis in the field, whether as a natural infection or as a biothreat remains challenging. Here we have developed a new lateral-flow immunochromatographic assay (LFIA) for B. anthracis spore detection based on the fact that conjugates of B. anthracis spores and super-paramagnetic particles labeled with antibodies will block the pores of chromatographic strips and form retention lines on the strips, instead of the conventionally reported test lines and control lines in classic LFIA. As a result, this new LFIA can simultaneously realize optical, magnetic and naked-eye detection by analyzing signals from the retention lines. As few as 500-700 pure B. anthracis spores can be recognized with CV values less than 8.31% within 5 min of chromatography and a total time of 20 min. For powdery sample tests, this LFIA can endure interference from 25% (w/v) milk, 10% (w/v) baking soda and 10% (w/v) starch without any sample pre-treatment, and has a corresponding detection limit of 6×10(4) spores/g milk powder, 2×10(5) spores/g starch and 5×10(5) spores/g baking soda. Compared with existing methods, this new approach is very competitive in terms of sensitivity, specificity, cost and ease of operation. This proof-of-concept study can also be extended for detection of many other large-sized analytes. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Lateral and vertical facies relationships of bedforms deposited by aggrading supercritical flows: From cyclic steps to humpback dunes

    NASA Astrophysics Data System (ADS)

    Lang, Jörg; Winsemann, Jutta

    2013-10-01

    The preservation of bedforms related to supercritical flows and hydraulic jumps is commonly considered to be rare in the geologic record, although these bedforms are known from a variety of depositional environments. This field-based study presents a detailed analysis of the sedimentary facies and stacking pattern of deposits of cyclic steps, chutes-and-pools, antidunes and humpback dunes from three-dimensional outcrops. The well exposed Middle Pleistocene successions from northern Germany comprise glacilacustrine ice-contact subaqueous fan and glacial lake-outburst flood deposits. The studied successions give new insights into the depositional architecture of bedforms related to supercritical flows and may serve as an analogue for other high-energy depositional environments such as fluvial settings, coarse-grained deltas or turbidite systems. Deposits of cyclic steps occur within the glacial lake-outburst flood succession and are characterised by lenticular scours infilled by gently to steeply dipping backsets. Cyclic steps formed due to acceleration and flow thinning when the glacial lake-outburst flood spilled over a push-moraine ridge. These bedforms are commonly laterally and vertically truncated and alternate with deposits of chutes-and-pools and antidunes. The subaqueous fan successions are dominated by laterally extensive sinusoidal waveforms, which are interpreted as deposits of aggrading stationary antidunes, which require quasi-steady flows at the lower limit of the supercritical flow stage and high rates of sedimentation. Humpback dunes are characterised by downflow divergent cross-stratification, displaying differentiation into topsets, foresets and bottomsets, and are interpreted as deposited at the transition from subcritical to supercritical flow conditions or vice versa. Gradual lateral and vertical transitions between humpback dunes and antidune deposits are very common. The absence of planar-parallel stratification in all studied successions

  2. VORSTAB: A computer program for calculating lateral-directional stability derivatives with vortex flow effect

    NASA Technical Reports Server (NTRS)

    Lan, C. Edward

    1985-01-01

    A computer program based on the Quasi-Vortex-Lattice Method of Lan is presented for calculating longitudinal and lateral-directional aerodynamic characteristics of nonplanar wing-body combination. The method is based on the assumption of inviscid subsonic flow. Both attached and vortex-separated flows are treated. For the vortex-separated flow, the calculation is based on the method of suction analogy. The effect of vortex breakdown is accounted for by an empirical method. A summary of the theoretical method, program capabilities, input format, output variables and program job control set-up are described. Three test cases are presented as guides for potential users of the code.

  3. Rapid Salmonella detection in experimentally inoculated equine faecal and veterinary hospital environmental samples using commercially available lateral flow immunoassays.

    PubMed

    Burgess, B A; Noyes, N R; Bolte, D S; Hyatt, D R; van Metre, D C; Morley, P S

    2015-01-01

    Salmonella enterica is the most commonly reported cause of outbreaks of nosocomial infections in large animal veterinary teaching hospitals and the closure of equine hospitals. Rapid detection may facilitate effective control practices in equine populations. Shipping and laboratory testing typically require ≥48 h to obtain results. Lateral flow immunoassays developed for use in food-safety microbiology provide an alternative that has not been evaluated for use with faeces or environmental samples. We aimed to identify enrichment methods that would allow commercially available rapid Salmonella detection systems (lateral flow immunoassays) to be used in clinical practice with equine faecal and environmental samples, providing test results in 18-24 h. In vitro experiment. Equine faecal and environmental samples were inoculated with known quantities of S. enterica serotype Typhimurium and cultured using 2 different enrichment techniques for faeces and 4 enrichment techniques for environmental samples. Samples were tested blindly using 2 different lateral flow immunoassays and plated on agar media for confirmatory testing. In general, commercial lateral flow immunoassays resulted in fewer false-negative test results with enrichment of 1 g faecal samples in tetrathionate for 18 h, while all environmental sample enrichment techniques resulted in similar detection rates. The limit of detection from spiked samples, ∼4 colony-forming units/g, was similar for all methods evaluated. The lateral flow immunoassays evaluated could reliably detect S. enterica within 18 h, indicating that they may be useful for rapid point-of-care testing in equine practice applications. Additional evaluation is needed using samples from naturally infected cases and the environment to gain an accurate estimate of test sensitivity and specificity and to substantiate further the true value of these tests in clinical practice. © 2014 EVJ Ltd.

  4. Liquid chromatography mass spectrometry determination of perfluoroalkyl acids in environmental solid extracts after phospholipid removal and on-line turbulent flow chromatography purification.

    PubMed

    Mazzoni, M; Polesello, S; Rusconi, M; Valsecchi, S

    2016-07-01

    An on-line TFC (Turbulent Flow Chromatography) clean up procedures coupled with UHPLC-MS/MS (Ultra High Performance Liquid Chromatography Mass Spectrometry) multi-residue method was developed for the simultaneous determination of 8 perfluroalkyl carboxylic acids (PFCA, from 5 to 12 carbon atoms) and 3 perfluoroalkyl sulfonic acids (PFSA, from 4 to 8 carbon atoms) in environmental solid matrices. Fast sample preparation procedure was based on a sonication-assisted extraction with acetonitrile. Phospholipids in biological samples were fully removed by an off-line SPE purification before injection, using HybridSPE(®) Phospholipid Ultra cartridges. The development of the on-line TFC clean-up procedure regarded the choice of the stationary phase, the optimization of the mobile phase composition, flow rate and injected volume. The validation of the optimized method included the evaluation of matrix effects, accuracy and reproducibility. Signal suppression in the analysis of fortified extracts ranged from 1 to 60%, and this problem was overcome by using isotopic dilution. Since no certified reference materials were available for PFAS in these matrices, accuracy was evaluated by recoveries on spiked clam samples which were 98-133% for PFCAs and 40-60% for PFSAs. MLDs and MLQs ranged from 0.03 to 0.3ngg(-1) wet weight and from 0.1 to 0.9ngg(-1) wet weight respectively. Repeatability (intra-day precision) and reproducibility (inter-day precision) showed RSD from 3 to 13% and from 4 to 27% respectively. Validated on-line TFC/UHPLC-MS/MS method has been applied for the determination of perfluoroalkyl acids in different solid matrices (sediment, fish, bivalves and bird yolk). Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Home-made online hyphenation of pressurized liquid extraction, turbulent flow chromatography, and high performance liquid chromatography, Cistanche deserticola as a case study.

    PubMed

    Song, Qingqing; Li, Jun; Liu, Xiao; Zhang, Yuan; Guo, Liping; Jiang, Yong; Song, Yuelin; Tu, Pengfei

    2016-03-18

    Incompatibility between the conventional pressurized liquid extraction (PLE) devices and high performance liquid chromatography (HPLC) extensively hinders direct and green chemical analysis of herbal materials. Herein, a facile PLE module was configured, and then it was online hyphenated with HPLC via a turbulent flow chromatography (TFC) column. Regarding PLE module, a long PEEK tube (0.13 × 1000 mm) was employed to generate desired pressure (approximately 13.0 MPa) when warm acidic water (70 °C) was delivered as extraction solvent at a high flow rate (2.5 mL/min), and a hollow guard column (3.0 × 4.0 mm) was implemented to hold crude materials. Effluent was collected from the outlet of PEEK tube, concentrated, and subjected onto HPLC coupled with hybrid ion trap-time of flight mass spectrometer to assess the extraction efficiency and also to profile the chemical composition of Cistanche deserticola (CD) that is honored as "Ginseng of the desert". Afterwards, a TFC column was introduced to accomplish online transmission of low molecule weight components from PLE module to HPLC coupled with diode array detection, and two electronic 6-port/2-channel valves were in charge of alternating the whole system between extraction (0-3.0 min) and elution (3.0-35.0 min) phases. Quantitative method was developed and validated for simultaneous determination of eight primary phenylethanoid glycosides in CD using online PLE-TFC-HPLC. All findings demonstrated that the home-made platform is advantageous at direct chemical analysis, as well as time-, solvent-, and material-savings, suggesting a robust tool for chemical fingerprinting of herbs. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Morphological resilience to flow fluctuations of fine sediment deposits in bank lateral cavities

    NASA Astrophysics Data System (ADS)

    Juez, C.; Thalmann, M.; Schleiss, A. J.; Franca, M. J.

    2018-05-01

    Lateral cavities are built in the banks of rivers for several purposes: to create harbors, to capture sediment, to keep a central navigable channel (i.e., Casiers de Girardon in the Rhone river) or to promote the formation of aquatic habitats if a limited amount of sediment is captured, providing hydraulic and morphologic diversity (i.e., the case of Japanese Wandos). This work is focused on this latter purpose: promotion of hydraulic and morphologic diversity. In these scenarios, an increase in the flow discharge in the main channel may, however, re-mobilize the deposit of sediment inside these lateral embayments and cause a sudden increase of the sediment concentration and turbidity in the main channel. It is thus of interest to characterize the resistance and resilience of these sedimentary deposits when the main channel is subjected to high flow or flushing events. Laboratory tests were carried out for five different normalized geometries of the cavities installed in the banks of an open channel and for five hydrographs with different levels of unsteadiness. Water depth, sediment deposit mass, sediment concentration and area covered by the settled sediments were recorded throughout each experiment. Although sediment deposits established at equilibrium before the flushing events are different depending on the geometry of the cavities, generally, they are recovered after being flushed by the high flow events. It is shown that the resistance and resilience of the sediment deposits are strongly dependent on the flow field and the mass exchange between the main channel and the cavities. This mass exchange is governed by the geometry of the cavities and the magnitude of the hydrographs applied.

  7. A fluorometric lateral flow assay for visual detection of nucleic acids using a digital camera readout.

    PubMed

    Magiati, Maria; Sevastou, Areti; Kalogianni, Despina P

    2018-06-04

    A fluorometric lateral flow assay has been developed for the detection of nucleic acids. The fluorophores phycoerythrin (PE) and fluorescein isothiocyanate (FITC) were used as labels, while a common digital camera and a colored vinyl-sheet, acting as a cut-off optical filter, are used for fluorescence imaging. After DNA amplification by polymerase chain reaction (PCR), the biotinylated PCR product is hybridized to its complementary probe that carries a poly(dA) tail at 3΄ edge and then applied to the lateral flow strip. The hybrids are captured to the test zone of the strip by immobilized poly(dT) sequences and detected by streptavidin-fluorescein and streptavidin-phycoerythrin conjugates, through streptavidin-biotin interaction. The assay is widely applicable, simple, cost-effective, and offers a large multiplexing potential. Its performance is comparable to assays based on the use of streptavidin-gold nanoparticles conjugates. As low as 7.8 fmol of a ssDNA and 12.5 fmol of an amplified dsDNA target were detectable. Graphical abstract Schematic presentation of a fluorometric lateral flow assay based on fluorescein and phycoerythrin fluorescent labels for the detection of single-stranded (ssDNA) and double-stranded DNA (dsDNA) sequences and using a digital camera readout. SA: streptavidin, BSA: Bovine Serum Albumin, B: biotin, FITC: fluorescein isothiocyanate, PE: phycoerythrin, TZ: test zone, CZ: control zone.

  8. A flow-through chromatography process for influenza A and B virus purification.

    PubMed

    Weigel, Thomas; Solomaier, Thomas; Peuker, Alessa; Pathapati, Trinath; Wolff, Michael W; Reichl, Udo

    2014-10-01

    Vaccination is still the most efficient measure to protect against influenza virus infections. Besides the seasonal wave of influenza, pandemic outbreaks of bird or swine flu represent a high threat to human population. With the establishment of cell culture-based processes, there is a growing demand for robust, economic and efficient downstream processes for influenza virus purification. This study focused on the development of an economic flow-through chromatographic process avoiding virus strain sensitive capture steps. Therefore, a three-step process consisting of anion exchange chromatography (AEC), Benzonase(®) treatment, and size exclusion chromatography with a ligand-activated core (LCC) was established, and tested for purification of two influenza A virus strains and one influenza B virus strain. The process resulted in high virus yields (≥68%) with protein contamination levels fulfilling requirements of the European Pharmacopeia for production of influenza vaccines for human use. DNA was depleted by ≥98.7% for all strains. The measured DNA concentrations per dose were close to the required limits of 10ng DNA per dose set by the European Pharmacopeia. In addition, the added Benzonase(®) could be successfully removed from the product fraction. Overall, the presented downstream process could potentially represent a simple, robust and economic platform technology for production of cell culture-derived influenza vaccines. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Conventional-Flow Liquid Chromatography-Mass Spectrometry for Exploratory Bottom-Up Proteomic Analyses.

    PubMed

    Lenčo, Juraj; Vajrychová, Marie; Pimková, Kristýna; Prokšová, Magdaléna; Benková, Markéta; Klimentová, Jana; Tambor, Vojtěch; Soukup, Ondřej

    2018-04-17

    Due to its sensitivity and productivity, bottom-up proteomics based on liquid chromatography-mass spectrometry (LC-MS) has become the core approach in the field. The de facto standard LC-MS platform for proteomics operates at sub-μL/min flow rates, and nanospray is required for efficiently introducing peptides into a mass spectrometer. Although this is almost a "dogma", this view is being reconsidered in light of developments in highly efficient chromatographic columns, and especially with the introduction of exceptionally sensitive MS instruments. Although conventional-flow LC-MS platforms have recently penetrated targeted proteomics successfully, their possibilities in discovery-oriented proteomics have not yet been thoroughly explored. Our objective was to determine what are the extra costs and what optimization and adjustments to a conventional-flow LC-MS system must be undertaken to identify a comparable number of proteins as can be identified on a nanoLC-MS system. We demonstrate that the amount of a complex tryptic digest needed for comparable proteome coverage can be roughly 5-fold greater, providing the column dimensions are properly chosen, extra-column peak dispersion is minimized, column temperature and flow rate are set to levels appropriate for peptide separation, and the composition of mobile phases is fine-tuned. Indeed, we identified 2 835 proteins from 2 μg of HeLa cells tryptic digest separated during a 60 min gradient at 68 μL/min on a 1.0 mm × 250 mm column held at 55 °C and using an aqua-acetonitrile mobile phases containing 0.1% formic acid, 0.4% acetic acid, and 3% dimethyl sulfoxide. Our results document that conventional-flow LC-MS is an attractive alternative for bottom-up exploratory proteomics.

  10. Rapid authentication of Cordyceps by lateral flow dipstick.

    PubMed

    Wong, Yuk-Lau; Wong, Ka-Lok; Shaw, Pang-Chui

    2015-01-01

    Cordyceps (Dongchongxiacao), a valuable traditional Chinese medicine, is composed of the fruiting body of Ophiocordyceps sinensis (Family: Ophiocordycipitaceae) on a caterpillar of ghost-moth species (Family: Hepialidae). Owing to its multiple potential functions, Cordyceps are in great demand and represent significant economic value. Adulterants or substitutes named Cordyceps or Chongcao from related fungi have been reported. In this study, polymerase chain reaction (PCR) coupled with a lateral flow dipstick (LFD) system was developed to distinguish genuine herb O. sinensis from its common adulterant Cordyceps gunnii and Cordyceps militaris. Specific primers (EF-CS-F1-Biotin, EF-CG-F1-Biotin and EF-CM-F1- Biotin) were designed to differentiate the three Cordyceps species. Internal control (EF-F1-b-DIG and EF-R1-FITC) was included to minimize the false signal due to PCR inhibitors or DNA degradation. LFD was then successfully employed for speedy and accurate detection of the respective PCR products. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Novel development of a lateral flow immunoassay for rapid field detection of citrus tristeza virus

    USDA-ARS?s Scientific Manuscript database

    Maintenance of virus-free citrus in nurseries and orchards is essential to control spread of aphid-borne Citrus tristeza virus (CTV) in California. A lateral flow assay (LFA) test strip with a polyclonal antiserum made from virus particles produced in Nicotiana benthamiana plants inoculated with an ...

  12. Analysis of bacterial fatty acids by flow modulated comprehensive two-dimensional gas chromatography with parallel flame ionization detector/mass spectrometry.

    PubMed

    Gu, Qun; David, Frank; Lynen, Frédéric; Rumpel, Klaus; Xu, Guowang; De Vos, Paul; Sandra, Pat

    2010-06-25

    Comprehensive two-dimensional gas chromatography (GCxGC) offers an interesting tool for profiling bacterial fatty acids. Flow modulated GCxGC using a commercially available system was evaluated, different parameters such as column flows and modulation time were optimized. The method was tested on bacterial fatty acid methyl esters (BAMEs) from Stenotrophomonas maltophilia LMG 958T by using parallel flame ionization detector (FID)/mass spectrometry (MS). The results are compared to data obtained using a thermal modulated GCxGC system. The data show that flow modulated GCxGC-FID/MS method can be applied in a routine environment and offers interesting perspectives for chemotaxonomy of bacteria.

  13. Evidence for lateral mantle plume flow feeding the Central Indian Ridge

    NASA Astrophysics Data System (ADS)

    Murton, B. J.; Tindle, A. G.

    2003-04-01

    The Central Indian Ridge exhibits morphological and geochemical features indicating lateral flow of shallow plume asthenosphere from the Reunion hot-spot to the ridge axis. South of the Marie Celeste fracture zone, at 18.25°S, the Central Indian Ridge is bound by a southward closing, “V”-shaped region of shallow crust that extends for over 800 km. Over this distance, the ridge axis deepens to the south and is also affected by left-stepping offsets that bring it towards the west. The northern end of the ridge, which is closest to the island of La'Réunion, is shallowest and dominated by an inflated segment with associated sheet flows covering over 50 square kilometres. These morphological features are usually associated with ridge-hot-spot interaction. However, the nearest active hot-spot lies over 1100 km to the west beneath the island of La'Réunion. Geochemical trends for basalts erupted along the Central Indian Ridge demonstrate a gradient of northward decreasing MgO and increasing SiO2, indicating a relationship between shallower crust and increased magmatic fractional crystallisation. Superimposed on this gradient is an excess increase in incompatible element ratios, indicative of mantle enrichment to the north. The enrichment correlates with the spreading-parallel distance between the ridge axis and the edge of the "V"-shaped region of anomalously shallow crust. Locally, the enriched mantle component is found preferentially at third-order ridge offsets and adjacent to the rift walls demonstrating melting of a compositionally stratified, spinel-lherzolite mantle. These features are evidence for shallow, lateral flow of enriched hot-spot asthenosphere at a velocity of ~333 mm yr-1 and with a flux of at least 50 m3 s-1, through a mantle 'worm', towards the ridge axis where it migrates south at a rate of 54 - 67 mm per year. The trend of the geochemical enrichment points to mixing between deeper N-MORB and shallower Reunion hot-spot sources beneath the

  14. Improved hydrophilic interaction chromatography LC/MS of heparinoids using a chip with postcolumn makeup flow.

    PubMed

    Staples, Gregory O; Naimy, Hicham; Yin, Hongfeng; Kileen, Kevin; Kraiczek, Karsten; Costello, Catherine E; Zaia, Joseph

    2010-01-15

    Heparan sulfate (HS) and heparin are linear, heterogeneous carbohydrates of the glycosaminoglycan (GAG) family that are modified by N-acetylation, N-sulfation, O-sulfation, and uronic acid epimerization. HS interacts with growth factors in the extracellular matrix, thereby modulating signaling pathways that govern cell growth, development, differentiation, proliferation, and adhesion. High-performance liquid chromatography (HPLC)-chip-based hydrophilic interaction liquid chromatography/mass spectrometry has emerged as a method for analyzing the domain structure of GAGs. However, analysis of highly sulfated GAG structures decasaccharide or larger in size has been limited by spray instability in the negative-ion mode. This report demonstrates that addition of postcolumn makeup flow to the amide-HPLC-chip configuration permits robust and reproducible analysis of extended GAG domains (up to degree of polymerization 18) from HS and heparin. This platform provides quantitative information regarding the oligosaccharide profile, degree of sulfation, and nonreducing chain termini. It is expected that this technology will enable quantitative, comparative glycomics profiling of extended GAG oligosaccharide domains of functional interest.

  15. Dielectrokinetic chromatography devices

    DOEpatents

    Chirica, Gabriela S; Fiechtner, Gregory J; Singh, Anup K

    2014-12-16

    Disclosed herein are methods and devices for dielectrokinetic chromatography. As disclosed, the devices comprise microchannels having at least one perturber which produces a non-uniformity in a field spanning the width of the microchannel. The interaction of the field non-uniformity with a perturber produces a secondary flow which competes with a primary flow. By decreasing the size of the perturber the secondary flow becomes significant for particles/analytes in the nanometer-size range. Depending on the nature of a particle/analyte present in the fluid and its interaction with the primary flow and the secondary flow, the analyte may be retained or redirected. The composition of the primary flow can be varied to affect the magnitude of primary and/or secondary flows on the particles/analytes and thereby separate and concentrate it from other particles/analytes.

  16. Study of flow fractionation characteristics of magnetic chromatography utilizing high-temperature superconducting bulk magnet.

    PubMed

    Fukui, Satoshi; Shoji, Yoshihiro; Ogawa, Jun; Oka, Tetsuo; Yamaguchi, Mitsugi; Sato, Takao; Ooizumi, Manabu; Imaizumi, Hiroshi; Ohara, Takeshi

    2009-02-01

    We present numerical simulation of separating magnetic particles with different magnetic susceptibilities by magnetic chromatography using a high-temperature superconducting bulk magnet. The transient transport is numerically simulated for two kinds of particles having different magnetic susceptibilities. The time evolutions were calculated for the particle concentration in the narrow channel of the spiral arrangement placed in the magnetic field. The field is produced by the highly magnetized high-temperature superconducting bulk magnet. The numerical results show the flow velocity difference of the particle transport corresponding to the difference in the magnetic susceptibility, as well as the possible separation of paramagnetic particles of 20 nm diameter.

  17. A trench study to assess transfer of pesticides in subsurface lateral flow for a soil with contrasting texture on a sloping vineyard in Beaujolais.

    PubMed

    Peyrard, X; Liger, L; Guillemain, C; Gouy, V

    2016-01-01

    Subsurface lateral flow in both texture-contrast soils and catchments with shallow bedrock is suspected to be a non-point source of contamination of watercourses by pesticides used in agriculture. As a case study, the north of the Beaujolais region (eastern France) provides a favorable environment for such contamination due to its agro-pedo-climatic conditions. Environments seen in the Beaujolais region include intense viticulture, permeable and shallow soils, steep hillslopes, and storms that occur during the periods of pesticide application. Watercourse contamination by pesticides has been widely observed in this region, and offsite pesticide transport by subsurface lateral flow is suspected to be involved in diffuse and chronic presence of pesticides in surface water. In order to confirm and quantify the potential role of such processes in pesticide transfer, an automated trench system has been designed. The trench was set up on a steep farmed hillslope in a texture-contrast soil. It was equipped with a tipping bucket flow meter and an automatic sampler to monitor pesticide concentrations in lateral flow at fine resolution, by means of a flow-dependent sampling strategy. Four pesticides currently used in vine growing were studied to provide a range of mobility properties: one insecticide (chlorpyrifos-methyl) and three fungicides (spiroxamine, tebuconazole, and dimethomorph). With this system, it was possible to study pesticide concentration dynamics in the subsurface lateral flow, generated by substantial rainfall events following pesticide applications. The experimental design ascertained to be a suitable method in which to monitor subsurface lateral flow and related transfer of pesticides.

  18. Imaging lateral groundwater flow in the shallow subsurface using stochastic temperature fields

    NASA Astrophysics Data System (ADS)

    Fairley, Jerry P.; Nicholson, Kirsten N.

    2006-04-01

    Although temperature has often been used as an indication of vertical groundwater movement, its usefulness for identifying horizontal fluid flow has been limited by the difficulty of obtaining sufficient data to draw defensible conclusions. Here we use stochastic simulation to develop a high-resolution image of fluid temperatures in the shallow subsurface at Borax Lake, Oregon. The temperature field inferred from the geostatistical simulations clearly shows geothermal fluids discharging from a group of fault-controlled hydrothermal springs, moving laterally through the subsurface, and mixing with shallow subsurface flow originating from nearby Borax Lake. This interpretation of the data is supported by independent geochemical and isotopic evidence, which show a simple mixing trend between Borax Lake water and discharge from the thermal springs. It is generally agreed that stochastic simulation can be a useful tool for extracting information from complex and/or noisy data and, although not appropriate in all situations, geostatistical analysis may provide good definition of flow paths in the shallow subsurface. Although stochastic imaging techniques are well known in problems involving transport of species, e.g. delineation of contaminant plumes from soil gas survey data, we are unaware of previous applications to the transport of thermal energy for the purpose of inferring shallow groundwater flow.

  19. Does the Alpha-defensin Immunoassay or the Lateral Flow Test Have Better Diagnostic Value for Periprosthetic Joint Infection? A Systematic Review.

    PubMed

    Eriksson, Hannah K; Nordström, Jakob; Gabrysch, Katja; Hailer, Nils P; Lazarinis, Stergios

    2018-05-01

    Measuring alpha-defensin concentrations in synovial fluid may help to diagnose periprosthetic joint infection (PJI). There are two commercially available methods for measuring alpha-defensin in synovial fluid: the enzyme-linked immunosorbent assay-based Synovasure® alpha-defensin immunoassay, which gives a numeric readout within 24 hours, and the Synovasure lateral flow test, which gives a binary readout within 20 minutes. There is no compilation of the existing literature to support the use of one of these two tests over the other. Does the immunoassay or the lateral flow test have better diagnostic value (sensitivity and specificity) in diagnosing PJI? We followed PRISMA guidelines and identified all studies on alpha-defensin concentration in synovial fluid as a PJI diagnostic marker, indexed to April 14, 2017, in PubMed, JSTOR, Google Scholar, and OVID databases. The search retrieved 1578 records. All prospective and retrospective studies on alpha-defensin as a PJI marker (PJI classified according to the criteria of the Musculoskeletal Infection Society) after THA or TKA were included in the analysis. All studies used only one of the two commercially available test methods, but none of them was comparative. After excluding studies with overlapping patient populations, four studies investigating the alpha-defensin immunoassay and three investigating the lateral flow test remained. Alpha-defensin immunoassay studies included 482 joints and lateral flow test studies included 119. The quality of the trials was assessed according to the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool. The heterogeneity among studies was evaluated by the I index, indicating that the heterogeneity of the included studies was low. Pooled sensitivity, specificity, positive and negative likelihood ratios, and receiver operating curves were calculated for each method and compared with each other. The alpha-defensin immunoassay had superior overall diagnostic value

  20. Rapid visual detection of cyprinid herpesvirus 2 by recombinase polymerase amplification combined with a lateral flow dipstick.

    PubMed

    Wang, H; Sun, M; Xu, D; Podok, P; Xie, J; Jiang, Ys; Lu, Lq

    2018-05-28

    Herpesviral haematopoietic necrosis (HVHN), caused by cyprinid herpesvirus 2 (CyHV-2), causes significant losses in crucian carp (Carassius carassius) aquaculture. Rapid and convenient DNA assay detection of CyHV-2 is useful for field diagnosis. Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that can amplify DNA within 30 min at ~37°C by simulating in vivo DNA recombination. Herein, a rapid and convenient detection assay based on RPA with a lateral flow dipstick (LFD) was developed for detecting CyHV-2. The highly conserved ORF72 of CyHV-2 was targeted by specific and sensitive primers and probes. The optimized assay takes only 15 min at 38°C using a water bath, with analysis of products by 2% agarose gel electrophoresis within 30 min. A simple lateral flow strip based on the unique probe in reaction buffer was developed for visualization. The entire RPA-LFD assay takes 50 min less than the routine PCR method, is 100 times more sensitive and displays no cross-reaction with other aquatic viruses. The combined isothermal RPA and lateral flow assay (RPA-LFD) provides a simple, rapid, reliable method that could improve field diagnosis of CyHV-2 when resources are limited. © 2018 John Wiley & Sons Ltd.

  1. The investigation of the lateral interaction effect's on traffic flow behavior under open boundaries

    NASA Astrophysics Data System (ADS)

    Bouadi, M.; Jetto, K.; Benyoussef, A.; El Kenz, A.

    2017-11-01

    In this paper, an open boundaries traffic flow system is studied by taking into account the lateral interaction with spatial defects. For a random defects distribution, if the vehicles velocities are weakly correlated, the traffic phases can be predicted by considering the corresponding inflow and outflow functions. Conversely, if the vehicles velocities are strongly correlated, a phase segregation appears inside the system's bulk which induces the maximum current appearance. Such velocity correlation depends mainly on the defects densities and the probabilities of lateral deceleration. However, for a compact defects distribution, the traffic phases are predictable by using the inflow in the system beginning, the inflow entering the defects zone and the outflow function.

  2. Multiplex Lateral Flow Immunoassays Based on Amorphous Carbon Nanoparticles for Detecting Three Fusarium Mycotoxins in Maize.

    PubMed

    Zhang, Xiya; Yu, Xuezhi; Wen, Kai; Li, Chenglong; Mujtaba Mari, Ghulam; Jiang, Haiyang; Shi, Weimin; Shen, Jianzhong; Wang, Zhanhui

    2017-09-13

    The detecting labels used for lateral flow immunoassays (LFAs) have been traditionally gold nanoparticles (GNPs) and, more recently, luminescent nanoparticles, such as quantum dots (QDs). However, these labels have low sensitivity and are costly, in particular, for trace detection of mycotoxins in cereals. Here, we provided a simple preparation procedure for amorphous carbon nanoparticles (ACNPs) and described multiplex LFAs employing ACNPs as labels (ACNP-LFAs) for detecting three Fusarium mycotoxins. The analytical performance of ACNPs in LFA was compared to GNPs and QDs using the same immunoreagents, except for the labels, allowing for their analytical characteristics to be objectively compared. The visual limit of detection for ACNP-LFAs in buffer was 8-fold better than GNPs and 2-fold better than QDs. Under optimized conditions, the quantitative limit of detection of ACNP-LFAs in maize was as low as 20 μg/kg for deoxynivalenol, 13 μg/kg for T-2 toxin, and 1 μg/kg for zearalenone. These measurements were much lower than the action level of these mycotoxins in maize. The accuracy and precision of the ACNP-LFAs were evaluated by analysis of spiked and incurred maize samples with recoveries of 84.6-109% and coefficients of variation below 13%. The results of ACNP-LFAs using naturally incurred maize samples showed good agreement with results from high-performance liquid chromatography-tandem mass spectrometry, indicating that ACNPs were more sensitive labels than and a promising alternative to GNPs used in LFAs for detecting mycotoxins in cereals.

  3. A lateral flow biosensor for detection of single nucleotide polymorphism by circular strand displacement reaction.

    PubMed

    Xiao, Zhuo; Lie, Puchang; Fang, Zhiyuan; Yu, Luxin; Chen, Junhua; Liu, Jie; Ge, Chenchen; Zhou, Xuemeng; Zeng, Lingwen

    2012-09-04

    A lateral flow biosensor for detection of single nucleotide polymorphism based on circular strand displacement reaction (CSDPR) has been developed. Taking advantage of high fidelity of T4 DNA ligase, signal amplification by CSDPR, and the optical properties of gold nanoparticles, this assay has reached a detection limit of 0.01 fM.

  4. Rapid and Sensitive Detection of Protein Biomarker Using a Portable Fluorescence Biosensor based on Quantum Dots and a Lateral Flow Test Strip

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Li, Zhaohui; Wang, Ying; Wang, Jun

    2010-08-15

    A portable fluorescence biosensor with rapid and ultrasensitive response for trace protein has been built up with quantum dots and lateral flow test strip. The superior signal brightness and high photostability of quantum dots are combined with the promising advantages of lateral flow test strip and resulted in high sensitivity, selectivity and speedy for protein detection. Nitrated ceruloplasmin, a significant biomarker for cardiovascular disease, lung cancer and stress response to smoking, was used as model protein to demonstrate the good performances of this proposed Qdot-based lateral flow test strip. Quantitative detection of nitrated ceruloplasmin was realized by recording the fluorescencemore » intensity of quantum dots captured on the test line. Under optimal conditions, this portable fluorescence biosensor displays rapid responses for nitrated ceruloplasmin in wide dynamic range with a detection limit of 0.1ng/mL (S/N=3). Furthermore, the biosensor was successfully utilized for spiked human plasma sample detection with the concentration as low as 1ng/mL. The results demonstrate that the quantum dot-based lateral flow test strip is capable for rapid, sensitive, and quantitative detection of nitrated ceruloplasmin and hold a great promise for point-of-care and in field analysis of other protein biomarkers.« less

  5. Alpha-defensin and the Synovasure lateral flow device for the diagnosis of prosthetic joint infection.

    PubMed

    Marson, B A; Deshmukh, S R; Grindlay, D J C; Scammell, B E

    2018-06-01

    Aims The aim of this review was to evaluate the available literature and to calculate the pooled sensitivity and specificity for the different alpha-defensin test systems that may be used to diagnose prosthetic joint infection (PJI). Materials and Methods Studies using alpha-defensin or Synovasure (Zimmer Biomet, Warsaw, Indiana) to diagnose PJI were identified from systematic searches of electronic databases. The quality of the studies was evaluated using the Quality Assessment of Studies of Diagnostic Accuracy (QUADAS) tool. Meta-analysis was completed using a bivariate model. Results A total of 11 eligible studies were included. The median QUADAS score was 13 (interquartile range 13 to 13) out of 14. Significant conflicts of interest were identified in five studies. The pooled sensitivity for the laboratory alpha-defensin test was 0.95 (95% confidence interval (CI) 0.91 to 0.98) and the pooled specificity was 0.97 (95% CI 0.95 to 0.98) for four studies with a threshold level of 5.2 mgl -1 The pooled sensitivity for the lateral flow cassette test was 0.85 (95% CI 0.74 to 0.92) and the pooled specificity was 0.90 (95% CI 0.91 to 0.98). There was a statistically significant difference in sensitivity (p = 0.019), but not specificity (p = 0.47). Conclusion Laboratory-based alpha-defensin testing remains a promising tool for diagnosing PJI. The lateral flow cassette has a significantly lower performance and pooled results are comparable to the leucocyte esterase test. Further studies are required before the widespread adoption of the lateral flow cassette alpha-defensin test. Cite this article: Bone Joint J 2018;100-B:703-11.

  6. The effect of gas and fluid flows on nonlinear lateral vibrations of rotating drill strings

    NASA Astrophysics Data System (ADS)

    Khajiyeva, Lelya; Kudaibergenov, Askar; Kudaibergenov, Askat

    2018-06-01

    In this work we develop nonlinear mathematical models describing coupled lateral vibrations of a rotating drill string under the effect of external supersonic gas and internal fluid flows. An axial compressive load and a torque also affect the drill string. The mathematical models are derived by the use of Novozhilov's nonlinear theory of elasticity with implementation of Hamilton's variation principle. Expressions for the gas flow pressure are determined according to the piston theory. The fluid flow is considered as added mass inside the curved tube of the drill string. Using an algorithm developed in the Mathematica computation program on the basis of the Galerkin approach and the stiffness switching method the numerical solution of the obtained approximate differential equations is found. Influences of the external loads, drill string angular speed of rotation, parameters of the gas and fluid flows on the drill string vibrations are shown.

  7. Evaluation of a lateral flow immunoassay for field identification of Solenopsis invicta (Hymenoptera: Formicidae) in Australia

    USDA-ARS?s Scientific Manuscript database

    In an effort to improve surveillance capacity for the exotic red imported fire ant, Solenopsis invicta, a lateral flow immunoassay (LFA) was recently evaluated by Biosecurity Queensland staff in Australia. The purpose of the research was to assess the ability of the fire ant LFA to discriminate S. i...

  8. The performance characteristics of lateral flow devices with 2 strains of highly pathogenic avian influenza virus

    USDA-ARS?s Scientific Manuscript database

    Lateral flow devices (LFD) are commercially available and provide a fast, highly specific, on-site test for avian influenza. Because of the low analytic sensitivity of LFD tests at low virus concentrations, targeted sampling of sick and dead birds has been proposed in order to increase detection pr...

  9. A high-resolution land model coupled with groundwater lateral flow, human water regulation and the changes in soil freeze-thaw fronts

    NASA Astrophysics Data System (ADS)

    Xie, Z.; Zeng, Y.; Liu, S.; Gao, J.; Jia, B.; Qin, P.

    2017-12-01

    Both anthropogenic water regulation and groundwater lateral flow essentially affect groundwater table patterns. Their relationship is close because lateral flow recharges the groundwater depletion cone, which is induced by over-exploitation. And the movement of frost and thaw fronts (FTFs) affects soil water and thermal characteristics, as well as energy and water exchanges between land surface and the atmosphere. In this study, schemes describing groundwater lateral flow, human water regulation and the changes in soil freeze-thaw fronts were developed and incorporated into the Community Land Model 4.5. Then the model was applied in Heihe River Basin(HRB), an arid and semiarid region, northwest China. High resolution ( 1 km) numerical simulations showed that groundwater lateral flow driven by changes in water heads can essentially change the groundwater table pattern with the deeper water table appearing in the hillslope regions and shallower water table appearing in valley bottom regions and plains. Over the last decade, anthropogenic groundwater exploitation deepened the water table by approximately 2 m in the middle reaches of the HRB and rapidly reduced the terrestrial water storage, while irrigation increased soil moisture by approximately 0.1 m3 m-3. The water stored in the mainstream of the Heihe River was also reduced by human surface water withdrawal. The latent heat flux was increased by 30 W m-2 over the irrigated region, with an identical decrease in sensible heat flux. The simulated groundwater lateral flow was shown to effectively recharge the groundwater depletion cone caused by over-exploitation. The offset rate is higher in plains than mountainous regions. In addition, the simulated FTFs depth compared well with the observed data both in D66 station (permafrost) and Hulugou station (seasonally frozen ground). Over the HRB, the upstream area is permafrost region with maximum thawed depth at 2.5 m and lower region is seasonal frozen ground region

  10. Isothermal Recombinase Polymerase amplification (RPA) of Schistosoma haematobium DNA and oligochromatographic lateral flow detection.

    PubMed

    Rosser, A; Rollinson, D; Forrest, M; Webster, B L

    2015-09-04

    Accurate diagnosis of urogenital schistosomiasis is vital for surveillance/control programs. Amplification of schistosome DNA in urine by PCR is sensitive and specific but requires infrastructure, financial resources and skilled personnel, often not available in endemic areas. Recombinase Polymerase Amplification (RPA) is an isothermal DNA amplification/detection technology that is simple, rapid, portable and needs few resources. Here a Schistosoma haematobium RPA assay was developed and adapted so that DNA amplicons could be detected using oligochromatographic Lateral Flow (LF) strips. The assay successfully amplified S. haematobium DNA at 30-45 °C in 10 mins and was sensitive to a lower limit of 100 fg of DNA. The assay was also successful with the addition of crude urine, up to 5% of the total reaction volume. Cross amplification occurred with other schistosome species but not with other common urine microorganisms. The LF-RPA assay developed here can amplify and detect low levels of S. haematobium DNA. Reactions are rapid, require low temperatures and positive reactions are interpreted using lateral flow strips, reducing the need for infrastructure and resources. This together with an ability to withstand inhibitors within urine makes RPA a promising technology for further development as a molecular diagnostic tool for urogenital schistosomiasis.

  11. Base flow recession from unsaturated-saturated porous media considering lateral unsaturated discharge and aquifer compressibility

    NASA Astrophysics Data System (ADS)

    Liang, Xiuyu; Zhan, Hongbin; Zhang, You-Kuan; Schilling, Keith

    2017-09-01

    Unsaturated flow is an important process in base flow recessions and its effect is rarely investigated. A mathematical model for a coupled unsaturated-saturated flow in a horizontally unconfined aquifer with time-dependent infiltrations is presented. The effects of the lateral discharge of the unsaturated zone and aquifer compressibility are specifically taken into consideration. Semianalytical solutions for hydraulic heads and discharges are derived using Laplace transform and Cosine transform. The solutions are compared with solutions of the linearized Boussinesq equation (LB solution) and the linearized Laplace equation (LL solution), respectively. A larger dimensionless constitutive exponent κD (a smaller retention capacity) of the unsaturated zone leads to a smaller discharge during the infiltration period and a larger discharge after the infiltration. The lateral discharge of the unsaturated zone is significant when κD≤1, and becomes negligible when κD≥100. The compressibility of the aquifer has a nonnegligible impact on the discharge at early times. For late times, the power index b of the recession curve -dQ/dt˜ aQb, is 1 and independent of κD, where Q is the base flow and a is a constant lumped aquifer parameter. For early times, b is approximately equal to 3 but it approaches infinity when t→0. The present solution is applied to synthetic and field cases. The present solution matched the synthetic data better than both the LL and LB solutions, with a minimum relative error of 16% for estimate of hydraulic conductivity. The present solution was applied to the observed streamflow discharge in Iowa, and the estimated values of the aquifer parameters were reasonable.

  12. Lateral traction of laminar flow between sliding pair with heterogeneous slip/no-slip surface

    NASA Astrophysics Data System (ADS)

    Wu, Zhenpeng; Zeng, Liangcai; Chen, Xiaolan; Chen, Keying; Ding, Xianzhong

    2017-11-01

    The problem of shaft axial motion which significantly affects the lubrication performance has been a common phenomenon in journal bearing systems. The existing work involved in the solution of shaft axial motion is also very rare. In this study, we choose to examine the flow between sliding pair in which regard we present a unique heterogeneous surface consisting of a slip zone and a no-slip zone. The results reveal the following points: 1) By appropriately arranging the slip zone to change the angle between the borderline and the moving direction of the upper plate, it is possible to control the direction of the lateral traction in which the liquid film acts on the upper plate. 2) Exponent of the power function of the borderline and aspect ratio of the computational domain are large or small are not conducive to increasing the effect of lateral traction. For the object of this study, the final results of the optimization are shown that the lateral traction can account for 20% of the resistance.

  13. Two-dimensional turbulent flow chromatography coupled on-line to liquid chromatography-mass spectrometry for solution-based ligand screening against multiple proteins.

    PubMed

    Zhou, Jian-Liang; An, Jing-Jing; Li, Ping; Li, Hui-Jun; Jiang, Yan; Cheng, Jie-Fei

    2009-03-20

    We present herein a novel bioseparation/chemical analysis strategy for protein-ligand screening and affinity ranking in compound mixtures, designed to increase screening rates and improve sensitivity and ruggedness in performance. The strategy is carried out by combining on-line two-dimensional turbulent flow chromatography (2D-TFC) with liquid chromatography-mass spectrometry (LC-MS), and accomplished through the following steps: (1) a reversed-phase TFC stage to separate the protein/ligand complex from the unbound free molecules, (2) an on-line dissociation process to release the bound ligands from the complexes, and (3) a second mixed-mode cation-exchange/reversed-phase TFC stage to trap the bound ligands and to remove the proteins and salts, followed by LC-MS analysis for identification and determination of the binding affinities. The technique can implement an ultra-fast isolation of protein/ligand complex with the retention time of a complex peak in about 5s, and on-line prepare the "clean" sample to be directly compatible with the LC-MS analysis. The improvement in performance of this 2D-TFC/LC-MS approach over the conventional approach has been demonstrated by determining affinity-selected ligands of the target proteins acetylcholinesterase and butyrylcholinesterase from a small library with known binding affinities and a steroidal alkaloid library composed of structurally similar compounds. Our results show that 2D-TFC/LC-MS is a generic and efficient tool for high-throughput screening of ligands with low-to-high binding affinities, and structure-activity relationship evaluation.

  14. Development of a split-flow system for high precision variable sample introduction in supercritical fluid chromatography.

    PubMed

    Sakai, Miho; Hayakawa, Yoshihiro; Funada, Yasuhiro; Ando, Takashi; Fukusaki, Eiichiro; Bamba, Takeshi

    2017-09-15

    In this study, we propose a novel variable sample injection system based on full-loop injection, named the split-flow sample introduction system, for application in supercritical fluid chromatography (SFC). In this system, the mobile phase is split by the differential pressure between two back pressure regulators (BPRs) after full-loop injection suitable for SFC, and this differential pressure determines the introduction rate. Nine compounds with a wide range of characteristics were introduced with high reproducibility and universality, confirming that a robust variable sample injection system was achieved. We also investigated the control factors of our proposed system. Sample introduction was controlled by the ratio between the column-side pressure drops in splitless and split flow, ΔP columnsideinsplitless and ΔP columnsideinsplit , respectively, where ΔP columnsideinsplitless is related to the mobile phase flow rate and composition and the column resistance. When all other conditions are kept constant, increasing the make-up flow induces an additional pressure drop on the column side of the system, which leads to a reduced column-side flow rate, and hence decreased the amount of sample injected, even when the net pressure drop on the column side remains the same. Thus, sample introduction could be highly controlled at low sample introduction rate, regardless of the introduction conditions. This feature is advantageous because, as a control factor, the solvent in the make-up pump is independent of the column-side pressure drop. Copyright © 2017. Published by Elsevier B.V.

  15. Improved liquid chromatography-MS/MS of heparan sulfate oligosaccharides via chip-based pulsed makeup flow.

    PubMed

    Huang, Yu; Shi, Xiaofeng; Yu, Xiang; Leymarie, Nancy; Staples, Gregory O; Yin, Hongfeng; Killeen, Kevin; Zaia, Joseph

    2011-11-01

    Microfluidic chip-based hydrophilic interaction chromatography (HILIC) is a useful separation system for liquid chromatography-mass spectrometry (LC-MS) in compositional profiling of heparan sulfate (HS) oligosaccharides; however, ions observed using HILIC LC-MS are low in charge. Tandem MS of HS oligosaccharide ions with low charge results in undesirable losses of SO(3) from precursor ions during collision induced dissociation. One solution is to add metal cations to stabilize sulfate groups. Another is to add a nonvolatile, polar compound such as sulfolane, a molecule known to supercharge proteins, to produce a similar effect for oligosaccharides. We demonstrate use of a novel pulsed makeup flow (MUF) HPLC-chip. The chip enables controlled application of additives during specified chromatographic time windows and thus minimizes the extent to which nonvolatile additives build up in the ion source. The pulsed MUF system was applied to LC-MS/MS of HS oligosaccharides. Metal cations and sulfolane were tested as additives. The most promising results were obtained for sulfolane, for which supercharging of the oligosaccharide ions increased their signal strengths relative to controls. Tandem MS of these supercharged precursor ions showed decreased abundances of product ions from sulfate losses yet more abundant product ions from backbone cleavages.

  16. Influence of lateral discomfort on the stability of traffic flow based on visual angle car-following model

    NASA Astrophysics Data System (ADS)

    Zheng, Liang; Zhong, Shiquan; Jin, Peter J.; Ma, Shoufeng

    2012-12-01

    Due to the poor road markings and irregular driving behaviors, not every vehicle is positioned in the center of the lane. The deviation from the center can cause discomfort to drivers in the neighboring lane, which is referred to as lateral discomfort (or lateral friction). Such lateral discomfort can be incorporated into the driver stimulus-response framework by considering the visual angle and its changing rate from the psychological viewpoint. In this study, a two-lane visual angle based car-following model is proposed and its stability condition is obtained through linear stability theory. Further derivations indicate that the neutral stability line of the model is asymmetry and four factors including the vehicle width and length, the lateral separation and the sensitivity regarding the changing rate of visual angle have large impacts on the stability of traffic flow. Numerical simulations further verify these theoretical results, and demonstrate that the behaviors of diverging, merging and lane changing can break the original steady state and cause traffic fluctuations. However, these fluctuations may be alleviated to some extent by reducing the lateral discomfort.

  17. A novel multi-walled carbon nanotube-based antibody conjugate for quantitative and semi-quantitative lateral flow assays.

    PubMed

    Sun, Wenjuan; Hu, Xiaolong; Liu, Jia; Zhang, Yurong; Lu, Jianzhong; Zeng, Libo

    2017-10-01

    In this study, the multi-walled carbon nanotubes (MWCNTs) were applied in lateral flow strips (LFS) for semi-quantitative and quantitative assays. Firstly, the solubility of MWCNTs was improved using various surfactants to enhance their biocompatibility for practical application. The dispersed MWCNTs were conjugated with the methamphetamine (MET) antibody in a non-covalent manner and then manufactured into the LFS for the quantitative detection of MET. The MWCNTs-based lateral flow assay (MWCNTs-LFA) exhibited an excellent linear relationship between the values of test line and MET when its concentration ranges from 62.5 to 1500 ng/mL. The sensitivity of the LFS was evaluated by conjugating MWCNTs with HCG antibody and the MWCNTs conjugated method is 10 times more sensitive than the one conjugated with classical colloidal gold nanoparticles. Taken together, our data demonstrate that MWCNTs-LFA is a more sensitive and reliable assay for semi-quantitative and quantitative detection which can be used in forensic analysis.

  18. Rapid Catalyst Screening by a Continuous-Flow Microreactor Interfaced with Ultra High Pressure Liquid Chromatography

    PubMed Central

    Fang, Hui; Xiao, Qing; Wu, Fanghui; Floreancig, Paul E.; Weber, Stephen G.

    2010-01-01

    A high-throughput screening system for homogeneous catalyst discovery has been developed by integrating a continuous-flow capillary-based microreactor with ultra-high pressure liquid chromatography (UHPLC) for fast online analysis. Reactions are conducted in distinct and stable zones in a flow stream that allows for time and temperature regulation. UHPLC detection at high temperature allows high throughput online determination of substrate, product, and byproduct concentrations. We evaluated the efficacies of a series of soluble acid catalysts for an intramolecular Friedel-Crafts addition into an acyliminium ion intermediate within one day and with minimal material investment. The effects of catalyst loading, reaction time, and reaction temperature were also screened. This system exhibited high reproducibility for high-throughput catalyst screening and allowed several acid catalysts for the reaction to be identified. Major side products from the reactions were determined through off-line mass spectrometric detection. Er(OTf)3, the catalyst that showed optimal efficiency in the screening, was shown to be effective at promoting the cyclization reaction on a preparative scale. PMID:20666502

  19. High-throughput and direct measurement of androgen levels using turbulent flow chromatography liquid chromatography-triple quadrupole mass spectrometry (TFC-LC-TQMS) to discover chemicals that modulate dihydrotestosterone production in human prostate cancer cells.

    PubMed

    Kang, Kyungsu; Peng, Lei; Jung, Yu-Jin; Kim, Joo Yeon; Lee, Eun Ha; Lee, Hee Ju; Kim, Sang Min; Sung, Sang Hyun; Pan, Cheol-Ho; Choi, Yongsoo

    2018-02-01

    To develop a high-throughput screening system to measure the conversion of testosterone to dihydrotestosterone (DHT) in cultured human prostate cancer cells using turbulent flow chromatography liquid chromatography-triple quadrupole mass spectrometry (TFC-LC-TQMS). After optimizing the cell reaction system, this method demonstrated a screening capability of 103 samples, including 78 single compounds and 25 extracts, in less than 12 h without manual sample preparation. Consequently, fucoxanthin, phenethyl caffeate, and Curcuma longa L. extract were validated as bioactive chemicals that inhibited DHT production in cultured DU145 cells. In addition, naringenin boosted DHT production in DU145 cells. The method can facilitate the discovery of bioactive chemicals that modulate the DHT production, and four phytochemicals are potential candidates of nutraceuticals to adjust DHT levels in male hormonal dysfunction.

  20. Novel approach based on one-tube nested PCR and a lateral flow strip for highly sensitive diagnosis of tuberculous meningitis.

    PubMed

    Sun, Yajuan; Chen, Jiajun; Li, Jia; Xu, Yawei; Jin, Hui; Xu, Na; Yin, Rui; Hu, Guohua

    2017-01-01

    Rapid and sensitive detection of Mycobacterium tuberculosis (M. Tb) in cerebrospinal fluid is crucial in the diagnosis of tuberculous meningitis (TBM), but conventional diagnostic technologies have limited sensitivity and specificity or are time-consuming. In this work, a novel, highly sensitive molecular diagnostic method, one-tube nested PCR-lateral flow strip test (OTNPCR-LFST), was developed for detecting M. tuberculosis. This one-tube nested PCR maintains the sensitivity of conventional two-step nested PCR and reduces both the chance of cross-contamination and the time required for analysis. The PCR product was detected by a lateral flow strip assay, which provided a basis for migration of the test to a point-of-care (POC) microfluidic format. The developed assay had an improved sensitivity compared with traditional PCR, and the limit of detection was up to 1 fg DNA isolated from M. tuberculosis. The assay was also specific for M. tuberculosis, and no cross-reactions were found in other non-target bacteria. The application of this technique to clinical samples was successfully evaluated, and OTNPCR-LFST showed 89% overall sensitivity and 100% specificity for TBM patients. This one-tube nested PCR-lateral flow strip assay is useful for detecting M. tuberculosis in TBM due to its rapidity, high sensitivity and simple manipulation.

  1. Novel approach based on one-tube nested PCR and a lateral flow strip for highly sensitive diagnosis of tuberculous meningitis

    PubMed Central

    Sun, Yajuan; Chen, Jiajun; Li, Jia; Xu, Yawei; Jin, Hui; Xu, Na; Yin, Rui

    2017-01-01

    Rapid and sensitive detection of Mycobacterium tuberculosis (M. Tb) in cerebrospinal fluid is crucial in the diagnosis of tuberculous meningitis (TBM), but conventional diagnostic technologies have limited sensitivity and specificity or are time-consuming. In this work, a novel, highly sensitive molecular diagnostic method, one-tube nested PCR-lateral flow strip test (OTNPCR-LFST), was developed for detecting M. tuberculosis. This one-tube nested PCR maintains the sensitivity of conventional two-step nested PCR and reduces both the chance of cross-contamination and the time required for analysis. The PCR product was detected by a lateral flow strip assay, which provided a basis for migration of the test to a point-of-care (POC) microfluidic format. The developed assay had an improved sensitivity compared with traditional PCR, and the limit of detection was up to 1 fg DNA isolated from M. tuberculosis. The assay was also specific for M. tuberculosis, and no cross-reactions were found in other non-target bacteria. The application of this technique to clinical samples was successfully evaluated, and OTNPCR-LFST showed 89% overall sensitivity and 100% specificity for TBM patients. This one-tube nested PCR-lateral flow strip assay is useful for detecting M. tuberculosis in TBM due to its rapidity, high sensitivity and simple manipulation. PMID:29084241

  2. Rapid determination of phenylethanolamine A in biological samples by enzyme-linked immunosorbent assay and lateral-flow immunoassay.

    PubMed

    Li, Xiangmei; Wang, Wenjun; Wang, Limiao; Wang, Qi; Pei, Xingyao; Jiang, Haiyang

    2015-10-01

    Phenylethanolamine A (PA) is a β-adrenergic agonist, which was first used in animal husbandry as a growth promoter in China in 2010. In this study, a monoclonal-antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) and lateral-flow immunoassay (LFA) for the detection of PA in swine urine and pork were developed. The immunogen was prepared by linking PA hapten with carrier protein via a diazotization method. The IC50 value of the optimized icELISA was 0.44 ng mL(-1). The limits of detection of the icELISA for PA in swine urine and pork were 0.13 ng mL(-1) and 0.39 ng g(-1), respectively. The recoveries of PA from spiked swine urine and pork were in the range 82.0-107.4 % and 81.8-113.3%, respectively, with the coefficients of variation in the range 4.1-16.2% and 1.2-6.3%, respectively. The mAbs had negligible cross reactivity with 10 other β-agonists. In contrast, the LFA had a cut-off level of 5 ng mL(-1) (g) in swine urine and pork, and the results could be achieved within 5 min. Ten blind samples of swine urine were analyzed simultaneously by icELISA, LFA, and ultra-high-performance liquid chromatography-tandem mass spectrometry, and the results of the three methods agreed well. Therefore, the combination of two immunoassays provides an effective and rapid screening method for detection of PA residues in biological samples.

  3. Measuring flow velocity and flow direction by spatial and temporal analysis of flow fluctuations.

    PubMed

    Chagnaud, Boris P; Brücker, Christoph; Hofmann, Michael H; Bleckmann, Horst

    2008-04-23

    If exposed to bulk water flow, fish lateral line afferents respond only to flow fluctuations (AC) and not to the steady (DC) component of the flow. Consequently, a single lateral line afferent can encode neither bulk flow direction nor velocity. It is possible, however, for a fish to obtain bulk flow information using multiple afferents that respond only to flow fluctuations. We show by means of particle image velocimetry that, if a flow contains fluctuations, these fluctuations propagate with the flow. A cross-correlation of water motion measured at an upstream point with that at a downstream point can then provide information about flow velocity and flow direction. In this study, we recorded from pairs of primary lateral line afferents while a fish was exposed to either bulk water flow, or to the water motion caused by a moving object. We confirm that lateral line afferents responded to the flow fluctuations and not to the DC component of the flow, and that responses of many fiber pairs were highly correlated, if they were time-shifted to correct for gross flow velocity and gross flow direction. To prove that a cross-correlation mechanism can be used to retrieve the information about gross flow velocity and direction, we measured the flow-induced bending motions of two flexible micropillars separated in a downstream direction. A cross-correlation of the bending motions of these micropillars did indeed produce an accurate estimate of the velocity vector along the direction of the micropillars.

  4. Dielectrokinetic chromatography and devices thereof

    DOEpatents

    Chirica, Gabriela S; Fiechtner, Gregory J; Singh, Anup K

    2014-04-22

    Disclosed herein are methods and devices for dielectrokinetic chromatography. As disclosed, the devices comprise microchannels having at least one perturber which produces a non-uniformity in a field spanning the width of the microchannel. The interaction of the field non-uniformity with a perturber produces a secondary flow which competes with a primary flow. By decreasing the size of the perturber the secondary flow becomes significant for particles/analytes in the nanometer-size range. Depending on the nature of a particle/analyte present in the fluid and its interaction with the primary flow and the secondary flow, the analyte may be retained or redirected. The composition of the primary flow can be varied to affect the magnitude of primary and/or secondary flows on the particles/analytes and thereby separate and concentrate it from other particles/analytes.

  5. Effect of reference conditions on flow rate, modifier fraction and retention in supercritical fluid chromatography.

    PubMed

    De Pauw, Ruben; Shoykhet Choikhet, Konstantin; Desmet, Gert; Broeckhoven, Ken

    2016-08-12

    When using compressible mobile phases such as fluidic CO2, the density, the volumetric flow rates and volumetric fractions are pressure dependent. The pressure and temperature definition of these volumetric parameters (referred to as the reference conditions) may alter between systems, manufacturers and operating conditions. A supercritical fluid chromatography system was modified to operate in two modes with different definition of the eluent delivery parameters, referred to as fixed and variable mode. For the variable mode, the volumetric parameters are defined with reference to the pump operating pressure and actual pump head temperature. These conditions may vary when, e.g. changing the column length, permeability, flow rate, etc. and are thus variable reference conditions. For the fixed mode, the reference conditions were set at 150bar and 30°C, resulting in a mass flow rate and mass fraction of modifier definition which is independent of the operation conditions. For the variable mode, the mass flow rate of carbon dioxide increases with system pump operating pressure, decreasing the fraction of modifier. Comparing the void times and retention factor shows that the deviation between the two modes is almost independent of modifier percentage, but depends on the operating pressure. Recalculating the set volumetric fraction of modifier to the mass fraction results in the same retention behaviour for both modes. This shows that retention in SFC can be best modelled using the mass fraction of modifier. The fixed mode also simplifies method scaling as it only requires matching average column pressure. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Evaluation of a new nanoparticle-based lateral-flow immunoassay for the exclusion of heparin-induced thrombocytopenia (HIT).

    PubMed

    Sachs, Ulrich J; von Hesberg, Jakob; Santoso, Sentot; Bein, Gregor; Bakchoul, Tamam

    2011-12-01

    Heparin-induced thrombocytopenia (HIT) is an adverse complication of heparin caused by HIT antibodies (abs) that recognise platelet factor 4-heparin (PF4/hep) complexes. Several laboratory tests are available for the confirmation and/or refutation of HIT. A reliable and rapid single-sample test is still pending. It was the objective of this study to evaluate a new lateral-flow immunoassay based on nanoparticle technology. A cohort of 452 surgical and medical patients suspected of having HIT was evaluated. All samples were tested in two IgG-specific ELISAs, in a particle gel immunoassay (PaGIA) and in a newly developed lateral-flow immunoassay (LFI-HIT) as well as in a functional test (HIPA). Clinical pre-test probability was determined using 4T's score. Platelet-activating antibodies were present in 34/452 patients, all of whom had intermediate to high clinical probability. PF4/hep abs were detected in 79, 87, 86, and 63 sera using the four different immunoassays. The negative predictive values (NPV) were 100% for both ELISA tests and LFI-HIT but only 99.2% for PaGIA. There were less false positives (n=29) in the LFI-HIT compared to any other test. Additionally, significantly less time was required to perform LFI-HIT than to perform the other immunoassays. In conclusion, a newly developed lateral-flow assay, LFI-HIT, was capable of identifying all HIT patients in a cohort in a short period of time. Beside an NPV of 100%, the rate of false-positive signals is significantly lower with LFI-HIT than with other immunoassay(s). These performance characteristics suggest a high potency in reducing the risk and costs in patients suspected of having HIT.

  7. Lateral Variability of Lava flow Morphologies in the Deccan Traps Large Igneous Province (India)

    NASA Astrophysics Data System (ADS)

    Vanderkluysen, L.; Rader, E. L.; Self, S.; Clarke, A. B.; Sheth, H.; Moyer, D. K.

    2016-12-01

    In continental flood basalt provinces (CFBs), lava flow morphologies have traditionally been classified in two distinct groups recognizable in the field, expressing two different modes of lava flow emplacement mechanisms: (a) compound lava flow fields dominated by meter-sized pāhoehoe toes and lobes; and (b) inflated sheet lobes tens to hundreds of meters in width and meters to tens of meters in height. Temporal transitions between these two emplacement styles have been recognized in many mafic large igneous provinces worldwide and seem to be a fundamental feature of CFBs. However, lateral variations in these morphologies remain poorly studied and understood. In the Deccan CFB of India, two principal hypotheses have been proposed to account for possible lateral variations in lava flow facies: that smaller toes and lobes occur in distal regions of flow fields, representing breakouts at the edges of larger inflated lavas; or on the contrary that smaller toes and lobes represent proximal facies. We conducted a field study focusing on two of the Deccan's formations, the Khandala and the Poladpur, located in the middle and upper sections of the province's defined chemostratigraphy. We studied nine sections along a 600 km long E-W transect, with the easternmost sections representing the most distal outcrops, ≥ 500 km away from inferred vents. The Khandala Formation is traditionally described as a sequence of three thick inflated sheet lobes in the well-exposed sections of the western Deccan. However, in the central Deccan, we find the Khandala to be much thicker overall, with half of its thickness dominated by small, meter-sized toes and lobes. Inflated sheet lobes of the Khandala are thinner on average in the central Deccan than further to the east or west. We document this transition as occurring progressively in outcrops only 80 km apart. In the Poladpur, the average thickness of inflated sheet lobes increases in distal outcrops of the eastern Deccan. We interpret

  8. Interaction Between Downwelling Flow and the Laterally-Varying Thickness of the North American Lithosphere Inferred from Seismic Anisotropy

    NASA Astrophysics Data System (ADS)

    Behn, M. D.; Conrad, C. P.; Silver, P. G.

    2005-12-01

    Shear flow in the asthenosphere tends to align olivine crystals in the direction of shear, producing a seismically anisotropic asthenosphere that can be detected using a number of seismic techniques (e.g., shear-wave splitting (SWS) and surface waves). In the ocean basins, where the asthenosphere has a relatively uniform thickness and lithospheric anisotropy appears to be small, observed azimuthal anisotropy is well fit by asthenospheric shear flow in global flow models driven by a combination of plate motions and mantle density heterogeneity. In contrast, beneath the continents both the lithospheric ceiling and asthenospheric thickness may vary considerably across cratonic regions and ocean-continent boundaries. To examine the influence of a continental lithosphere with variable thickness on predictions of continental seismic anisotropy, we impose lateral variations in lithospheric viscosity in global models of mantle flow driven by plate motions and mantle density heterogeneity. For the North American continent, the Farallon slab descends beneath a deep cratonic root, producing downwelling flow in the upper mantle and convergent flow beneath the cratonic lithosphere. We evaluate both the orientation of the predicted azimuthal anisotropy and the depth dependence of radial anisotropy for this downwelling flow and find that the inclusion of a strong continental root provides an improved fit to observed SWS observations beneath the North American craton. Thus, we hypothesize that at least some continental anisotropy is associated with sub-lithospheric viscous shear, although fossil anisotropy in the lithospheric layer may also contribute significantly. Although we do not observe significant variations in the direction of predicted anisotropy with depth, we do find that the inclusion of deep continental roots pushes the depth of the anisotropy layer deeper into the upper mantle. We test several different models of laterally-varying lithosphere and asthenosphere

  9. Inferring common cognitive mechanisms from brain blood-flow lateralization data: a new methodology for fTCD analysis.

    PubMed

    Meyer, Georg F; Spray, Amy; Fairlie, Jo E; Uomini, Natalie T

    2014-01-01

    Current neuroimaging techniques with high spatial resolution constrain participant motion so that many natural tasks cannot be carried out. The aim of this paper is to show how a time-locked correlation-analysis of cerebral blood flow velocity (CBFV) lateralization data, obtained with functional TransCranial Doppler (fTCD) ultrasound, can be used to infer cerebral activation patterns across tasks. In a first experiment we demonstrate that the proposed analysis method results in data that are comparable with the standard Lateralization Index (LI) for within-task comparisons of CBFV patterns, recorded during cued word generation (CWG) at two difficulty levels. In the main experiment we demonstrate that the proposed analysis method shows correlated blood-flow patterns for two different cognitive tasks that are known to draw on common brain areas, CWG, and Music Synthesis. We show that CBFV patterns for Music and CWG are correlated only for participants with prior musical training. CBFV patterns for tasks that draw on distinct brain areas, the Tower of London and CWG, are not correlated. The proposed methodology extends conventional fTCD analysis by including temporal information in the analysis of cerebral blood-flow patterns to provide a robust, non-invasive method to infer whether common brain areas are used in different cognitive tasks. It complements conventional high resolution imaging techniques.

  10. Effect of geometry and scale for axial and radial flow membrane chromatography-Experimental study of bovin serum albumin adsorption.

    PubMed

    Teepakorn, Chalore; Fiaty, Koffi; Charcosset, Catherine

    2015-07-17

    During the last 10 years, membrane chromatography (MC) has been increasingly reported for biomolecule purification at both small and large scales. Although, several axial and radial flow MC devices are commercialized, the effect of the device dimensions on the adsorption performance has not been fully investigated. In this study, axial and radial flow anion ion-exchange MC devices were used for bovine serum albumin (BSA) adsorption. For both axial and radial flow, three devices at different scales were compared, two having similar diameter and two similar bed height. The pressure drop and the flow distribution using acetone as a non-binding solute were measured, as well as BSA breakthrough curves at different flow rates and BSA loading concentrations. For all devices, it was observed that the flow rate had no effect on the breakthrough curve, which confirms the advantage of MC to be used at high flow rates. In addition, the BSA binding capacity increased with increasing BSA concentration, which suggests that it could be preferable to work with concentrated solutions rather than with very dilute solutions, when using buffer at high phosphate concentration. For both axial and radial flow, the bed height had a negative impact on the binding capacity, as the lowest binding capacities per membrane volume were obtained with the devices having the highest bed height. Radial flow MC has potential at large-scale applications, as a short bed thickness can be combined with a large inlet surface area. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Investigation of drag and heat reduction induced by a novel combinational lateral jet and spike concept in supersonic flows based on conjugate heat transfer approach

    NASA Astrophysics Data System (ADS)

    Zhu, Liang; Chen, Xiong; Li, Yingkun; Musa, Omer; Zhou, Changsheng

    2018-01-01

    When flying at supersonic or hypersonic speeds through the air, the drag and severe heating have a great impact on the vehicles, thus the drag reduction and thermal protection studies have attracted worldwide attention. In the current study, the Reynolds-averaged Navier-Stokes (RANS) equations coupled with the shear stress transport (SST) k - ω turbulence model have been employed to investigate the flow behavior induced by a novel combinational lateral jet and spike concept in supersonic flows. A coupling conjugate heat transfer (CHT) approach has been applied to investigate the thermal protection, which takes the heat transfer of structure into consideration. After the code was validated by the available experimental results and the gird independency analysis was carried out, the influences of the spike length ratio, lateral jet pressure ratio and lateral jet location on the drag and heat reduction performance are analyzed comprehensively. The obtained results show that a remarkable reduction in the drag and heat flux is achieved when a lateral jet is added to the spike. This implies that the combinational lateral jet and spike concept in supersonic flows have a great benefit to the drag and heat reduction. Both the drag and heat reduction decrease with the increase of the lateral jet pressure ratio, and the heat flux is more sensitive to the lateral jet pressure ratio. The lateral jet should not be located in the bottom of the spike in order to realize better drag and heat reduction performance. The drag and heat flux could be reduced by about 45% by reasonable lateral jet location. The drag decreases with the increase of the spike length ratio whereas the heat flux is affected by the spike length ratio just in a certain range.

  12. Leading-edge flow reattachment and the lateral static stability of low-aspect-ratio rectangular wings

    NASA Astrophysics Data System (ADS)

    Linehan, Thomas; Mohseni, Kamran

    2017-11-01

    The relationship between lateral static stability derivative, Clβ,lift coefficient, CL, and angle of attack was investigated for rectangular wings of aspect ratio A R =0.75 ,1 ,1.5 , and 3 using Stereo-Digital Particle Image Velocimetry (S-DPIV) and direct force and moment measurements. When the product Cl βA R is plotted with respect to CL, the lateral stability curves of each wing collapse to a single line for CL<0.7 . For CL>0.7 , the linearity and scaling of Clβwith respect to CL is lost. S-DPIV is used to elucidate the flow physics in this nonlinear regime. At α =10∘ , the leading-edge separation region emerges on the leeward portion of the sideslipped wing by means of vortex shedding. For the A R ≤1.5 wings at α >15∘ , the tip vortex downwash is sufficient to restrict the shedding of leading-edge vorticity thereby sustaining the lift of the leading-edge separation region at high angles of attack. Concurrently, the windward tip vortex grows in size and strength with increasing angle of attack, displacing the leading-edge separation region further toward the leeward wing. This reorganization of lift-generating vorticity results in the initial nonlinearities between Cl β and CL at angles of attack for which CL is still increasing. At angles of attack near that of maximum lift for the A R ≤1 wings, the windward tip vortex lifts off the wing, decreasing the lateral static stability of the wing prior to lift stall. For the A R =3 wing at α >10∘ , nonlinear trends in Cl β versus CL occur due to the spanwise evolution of stalled flow.

  13. Gas-liquid flow splitting in T-junction with inclined lateral arm

    NASA Astrophysics Data System (ADS)

    Yang, Le-le; Liu, Shuo; Li, Hua; Zhang, Jian; Wu, Ying-xiang; Xu, Jing-yu

    2018-02-01

    This paper studies the gas-liquid flow splitting in T-junction with inclined lateral arm. The separation mechanism of the T-junction is related to the pressure distribution in the T-junction. It is shown that the separation efficiency strongly depends on the inclination angle, when the angle ranges from 0° to 30°, while not so strongly for angles in the range from 30° to 90° Increasing the number of connecting tubes is helpful for the gas-liquid separation, and under the present test conditions, with four connecting tubes, a good separation performance can be achieved. Accordingly, a multi-tube Y-junction separator with four connecting tubes is designed for the experimental investigation. A good agreement between the simulated and measured data shows that there is an optimal split ratio to achieve the best performance for the multi-tube Y-junction separator.

  14. Electrospin-coating of nitrocellulose membrane enhances sensitivity in nucleic acid-based lateral flow assay.

    PubMed

    Yew, Chee-Hong Takahiro; Azari, Pedram; Choi, Jane Ru; Li, Fei; Pingguan-Murphy, Belinda

    2018-06-07

    Point-of-care biosensors are important tools developed to aid medical diagnosis and testing, food safety and environmental monitoring. Paper-based biosensors, especially nucleic acid-based lateral flow assays (LFA), are affordable, simple to produce and easy to use in remote settings. However, the sensitivity of such assays to infectious diseases has always been a restrictive challenge. Here, we have successfully electrospun polycaprolactone (PCL) on nitrocellulose (NC) membrane to form a hydrophobic coating to reduce the flow rate and increase the interaction rate between the targets and gold nanoparticles-detecting probes conjugates, resulting in the binding of more complexes to the capture probes. With this approach, the sensitivity of the PCL electrospin-coated test strip has been increased by approximately ten-fold as compared to the unmodified test strip. As a proof of concept, this approach holds great potential for sensitive detection of targets at point-of-care testing. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Schinus terebinthifolius countercurrent chromatography (Part III): Method transfer from small countercurrent chromatography column to preparative centrifugal partition chromatography ones as a part of method development.

    PubMed

    das Neves Costa, Fernanda; Hubert, Jane; Borie, Nicolas; Kotland, Alexis; Hewitson, Peter; Ignatova, Svetlana; Renault, Jean-Hugues

    2017-03-03

    Countercurrent chromatography (CCC) and centrifugal partition chromatography (CPC) are support free liquid-liquid chromatography techniques sharing the same basic principles and features. Method transfer has previously been demonstrated for both techniques but never from one to another. This study aimed to show such a feasibility using fractionation of Schinus terebinthifolius berries dichloromethane extract as a case study. Heptane - ethyl acetate - methanol -water (6:1:6:1, v/v/v/v) was used as solvent system with masticadienonic and 3β-masticadienolic acids as target compounds. The optimized separation methodology previously described in Part I and II, was scaled up from an analytical hydrodynamic CCC column (17.4mL) to preparative hydrostatic CPC instruments (250mL and 303mL) as a part of method development. Flow-rate and sample loading were further optimized on CPC. Mobile phase linear velocity is suggested as a transfer invariant parameter if the CPC column contains sufficient number of partition cells. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Determination of free cortisol and free cortisone in human urine by on-line turbulent flow chromatography coupled to fused-core chromatography-tandem mass spectrometry (TFC-HPLC-MS/MS).

    PubMed

    Sánchez-Guijo, Alberto; Hartmann, Michaela F; Shi, Lijie; Remer, Thomas; Wudy, Stefan A

    2014-01-01

    Urinary free cortisol and urinary free cortisone are decisive markers for the diagnosis of syndromes related to the dysfunction of the adrenal gland or to evaluate certain enzymatic disorders. Here, we present a new method, designed for routine laboratory use, which enables quick determination of these analytes with minor sample workup. Turbulent flow chromatography shortens sample preparation, and connection to a fused-core particle-packed column (rugged amide-embedded C18 phase) permits a rapid and effective separation of the analytes, as well as additional separation from other related and isobaric compounds present in urine. Urinary isobaric compounds were successfully identified. The method requires only 100 μl of urine supernatant per sample. The total time between injections is 9.5 min. The solvents used for both turbulent and analytical chromatography are water and methanol, and the relatively low flows needed during the method resulted in an extended life of the columns. Linearity showed a R (2) > 0.994. Limit of detection and limit of quantification are 0.5 and 1.0 ng/ml for cortisone and 1.0 and 2.0 ng/ml for cortisol. Recoveries ranged from 99.7 to 109.1 % for cortisone and from 98.7 to 102.9 % for cortisol. Accuracy values (relative errors) for intra- and inter-assay experiments were always below 8 %, whereas precision (percent CV) ranged from 3.7 to 10.7 %. No matrix effects were detected during the validation process. The reproducibility for each analyte's retention time was excellent, with a coefficient of variation always below 0.2 %. The final validation step included the study of urine samples from healthy children and from children previously diagnosed with corticoidal disorders. The high selectivity achieved enables quick data handling.

  17. Gold nanocage-based lateral flow immunoassay for immunoglobulin G

    PubMed Central

    Yang, Yunhui; Ozsoz, Mehmet

    2017-01-01

    The authors describe a gold nanocage-based lateral flow strip biosensor (LFSB) for low-cost and sensitive detection of IgG. This protein was used as a model analyte to demonstrate the proof-of-concept. The method combines the unique optical properties of gold nanocages (GNCs) with highly efficient chromatographic separation. A sandwich-type of immunoreactions occurs on the GNC-based LFSB which has the attractive features of avoiding multiple incubation, separation, and washing steps. The captured GNCs on the purple test zone and control zone of the biosensor are producing characteristic purple bands, and this enables IgG even to be visually detected. Quantitatation was accomplished by reading the intensities of the bands with a portable strip reader. The LFSB fabrication and assay parameters were optimized. The biosensor displays a linear response in the 0.5 to 50 ng·mL−1 IgG concentration range, and it has a 15 min assay time. The detection limit is 0.1 ng·mL−1 of IgG, which is 2.5 times lower than that when using a gold nanoparticle-based LFSB. In our perception, this assay has a wide potential for the detection of other proteins and species for which respective antibodies are available. PMID:29187761

  18. Nitrate removal with lateral flow sulphur autotrophic denitrification reactor.

    PubMed

    Lv, Xiaomei; Shao, Mingfei; Li, Ji; Xie, Chuanbo

    2014-01-01

    An innovative lateral flow sulphur autotrophic denitrification (LFSAD) reactor was developed in this study; the treatment performance was evaluated and compared with traditional sulphur/limestone autotrophic denitrification (SLAD) reactor. Results showed that nitrite accumulation in the LFSAD reactor was less than 1.0 mg/L during the whole operation. Denitrification rate increased with the increased initial alkalinity and was approaching saturation when initial alkalinity exceeded 2.5 times the theoretical value. Higher influent nitrate concentration could facilitate nitrate removal capacity. In addition, denitrification efficiency could be promoted under an appropriate reflux ratio, and the highest nitrate removal percentage was achieved under reflux ratio of 200%, increased by 23.8% than that without reflux. Running resistance was only about 1/9 of that in SLAD reactor with equal amount of nitrate removed, which was the prominent excellence of the new reactor. In short, this study indicated that the developed reactor was feasible for nitrate removal from waters with lower concentrations, including contaminated surface water, groundwater or secondary effluent of municipal wastewater treatment with fairly low running resistance. The innovation in reactor design in this study may bring forth new ideas of reactor development of sulphur autotrophic denitrification for nitrate-contaminated water treatment.

  19. Experimental consideration of capillary chromatography based on tube radial distribution of ternary mixture carrier solvents under laminar flow conditions.

    PubMed

    Jinno, Naoya; Hashimoto, Masahiko; Tsukagoshi, Kazuhiko

    2011-01-01

    A capillary chromatography system has been developed based on the tube radial distribution of the carrier solvents using an open capillary tube and a water-acetonitrile-ethyl acetate mixture carrier solution. This tube radial distribution chromatography (TRDC) system works under laminar flow conditions. In this study, a phase diagram for the ternary mixture carrier solvents of water, acetonitrile, and ethyl acetate was constructed. The phase diagram that included a boundary curve between homogeneous and heterogeneous solutions was considered together with the component ratios of the solvents in the homogeneous carrier solutions required for the TRDC system. It was found that the TRDC system performed well with homogeneous solutions having component ratios of the solvents that were positioned near the homogeneous-heterogeneous solution boundary of the phase diagram. For preparing the carrier solutions of water-hydrophilic/hydrophobic organic solvents for the TRDC system, we used for the first time methanol, ethanol, 1,4-dioxane, and 1-propanol, instead of acetonitrile (hydrophilic organic solvent), as well as chloroform and 1-butanol, instead of ethyl acetate (hydrophobic organic solvent). The homogeneous ternary mixture carrier solutions were prepared near the homogeneous-heterogeneous solution boundary. Analyte mixtures of 2,6-naphthalenedisulfonic acid and 1-naphthol were separated with the TRDC system using these homogeneous ternary mixture carrier solutions. The pressure change in the capillary tube under laminar flow conditions might alter the carrier solution from homogeneous in the batch vessel to heterogeneous, thus affecting the tube radial distribution of the solvents in the capillary tube.

  20. Characterization of a protein conjugate using an asymmetrical-flow field-flow fractionation and a size-exclusion chromatography with multi-detection system.

    PubMed

    Rebolj, Katja; Pahovnik, David; Zagar, Ema

    2012-09-04

    In this study we present detailed characterization of a protein-PEG conjugate using two separation techniques, that is, asymmetrical-flow field-flow fractionation (AF4) and size-exclusion chromatography (SEC), which were online coupled to a series of successively connected detectors: an ultraviolet, a multiangle light-scattering, a quasi-elastic light-scattering, and a refractive-index detector (UV-MALS(QELS)-RI). Matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used as a complementary characterization technique. The results of AF4 as well as SEC on two columns connected in series, with both separation techniques coupled to a multidetection system, indicate the uniform molar mass and chemical composition of the conjugate, that is, the molar ratio of protein to PEG is 1/1, the presence of minute amounts of residual unreacted protein and the aggregates with the same chemical composition as that of the conjugate. Since the portion of aggregated species is smaller in the acetate buffer solution containing 5% sorbitol than in the acetate buffer solution with 200-mM sodium chloride, the former buffer solution is more suitable for conjugate storage. The separation using only one SEC column results in poorly resolved peaks of the PEGylated protein conjugate and the aggregates, whereas MALDI-TOF MS analysis reveal the presence of the residual protein, but not the aggregates.

  1. Energy measurement using flow computers and chromatography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Beeson, J.

    1995-12-01

    Arkla Pipeline Group (APG), along with most transmission companies, went to electronic flow measurement (EFM) to: (1) Increase resolution and accuracy; (2) Real time correction of flow variables; (3) Increase speed in data retrieval; (4) Reduce capital expenditures; and (5) Reduce operation and maintenance expenditures Prior to EFM, mechanical seven day charts were used which yielded 800 pressure and differential pressure readings. EFM yields 1.2-million readings, a 1500 time improvement in resolution and additional flow representation. The total system accuracy of the EFM system is 0.25 % compared with 2 % for the chart system which gives APG improved accuracy.more » A typical APG electronic measurement system includes a microprocessor-based flow computer, a telemetry communications package, and a gas chromatograph. Live relative density (specific gravity), BTU, CO{sub 2}, and N{sub 2} are updated from the chromatograph to the flow computer every six minutes which provides accurate MMBTU computations. Because the gas contract length has changed from years to monthly and from a majority of direct sales to transports both Arkla and its customers wanted access to actual volumes on a much more timely basis than is allowed with charts. The new electronic system allows volumes and other system data to be retrieved continuously, if EFM is on Supervisory Control and Data Acquisition (SCADA) or daily if on dial up telephone. Previously because of chart integration, information was not available for four to six weeks. EFM costs much less than the combined costs of telemetry transmitters, pressure and differential pressure chart recorders, and temperature chart recorder which it replaces. APG will install this equipment on smaller volume stations at a customers expense. APG requires backup measurement on metering facilities this size. It could be another APG flow computer or chart recorder, or the other companies flow computer or chart recorder.« less

  2. Internal flows and force matrices in axial flow inducers

    NASA Astrophysics Data System (ADS)

    Bhattacharyya, Abhijit

    1994-01-01

    Axial flow inducers such as those used in high speed rocket engine turbopumps are subject to complex internal flows and fluid-induced lateral and rotordynamic forces. An investigation of these internal flows was conducted using boundary layer flow visualization on the blades, hub and housing of unshrouded and shrouded inducers. Results showed that the blade boundary layer flows have strong radial components at off-design conditions and remain attached to the blade surface at all flow coefficients tested. The origin of upstream swirling backflow was found to be at the discharge plane of the inducer. In addition, flow reversal was observed at the suction side blade tip near the leading edge in a shrouded inducer. Re-entry of the hub boundary layer flow, a downstream backflow, into the blade passage area was observed at flow coefficients below design. For unshrouded inducers the radially outward flow near the blade tip mixed with the leakage flow to form the upstream backflow. The lateral and rotordynamic forces acting on an inducer due to an imposed whirl motion was also investigated at various flow coefficients. It was found that the rotordynamic force data at various whirl frequency ratios does not allow a normal quadratic fit; consequently the conventional inertial, stiffness and damping coefficients cannot be obtained and a definite whirl ratio describing the instability region does not result. Application of an actuator disk theory proved to be inaccurate in estimating the rotordynamic tangential force in a non-whirling inducer. The effect of upstream and downstream flow distortions on the rotordynamic and lateral forces on an inducer were studied. It was found that at flow coefficients below design, large lateral forces occurred in the presence of a downstream asymmetry. Results of inlet distortion experiments show that a strong inlet shear causes a significant increase in the lateral force. Cavitation was found to have important consequences for fluid

  3. Proteomic Identification of Immunodiagnostic Antigens for Trypanosoma vivax Infections in Cattle and Generation of a Proof-of-Concept Lateral Flow Test Diagnostic Device.

    PubMed

    Fleming, Jennifer R; Sastry, Lalitha; Wall, Steven J; Sullivan, Lauren; Ferguson, Michael A J

    2016-09-01

    Trypanosoma vivax is one of the causative agents of Animal African Trypanosomosis in cattle, which is endemic in sub-Saharan Africa and transmitted primarily by the bite of the tsetse fly vector. The parasite can also be mechanically transmitted, and this has allowed its spread to South America. Diagnostics are limited for this parasite and in farm settings diagnosis is mainly symptom-based. We set out to identify, using a proteomic approach, candidate diagnostic antigens to develop into an easy to use pen-side lateral flow test device. Two related members the invariant surface glycoprotein family, TvY486_0045500 and TvY486_0019690, were selected. Segments of these antigens, lacking N-terminal signal peptides and C-terminal transmembrane domains, were expressed in E. coli. Both were developed into ELISA tests and one of them, TvY486_0045500, was developed into a lateral flow test prototype. The tests were all evaluated blind with 113 randomised serum samples, taken from 37 calves before and after infection with T. vivax or T. congolense. The TvY486_0045500 and TvY486_0019690 ELISA tests gave identical sensitivity and specificity values for T. vivax infection of 94.5% (95% CI, 86.5% to 98.5%) and 88.0% (95% CI, 75.7% to 95.5%), respectively, and the TvY486_0045500 lateral flow test prototype a sensitivity and specificity of 92.0% (95% CI, 83.4% to 97.0%) and 89.8% (95% CI, 77.8% to 96.6%), respectively. These data suggest that recombinant TvY486_0045500 shows promise for the development of a pen-side lateral flow test for the diagnosis of T. vivax animal African trypanosomosis.

  4. Lipidomics by Supercritical Fluid Chromatography

    PubMed Central

    Laboureur, Laurent; Ollero, Mario; Touboul, David

    2015-01-01

    This review enlightens the role of supercritical fluid chromatography (SFC) in the field of lipid analysis. SFC has been popular in the late 1980s and 1990s before almost disappearing due to the commercial success of liquid chromatography (LC). It is only 20 years later that a regain of interest appeared when new commercial instruments were introduced. As SFC is fully compatible with the injection of extracts in pure organic solvent, this technique is perfectly suitable for lipid analysis and can be coupled with either highly universal (UV or evaporative light scattering) or highly specific (mass spectrometry) detection methods. A short history of the use of supercritical fluids as mobile phase for the separation oflipids will be introduced first. Then, the advantages and drawbacks of SFC are discussed for each class of lipids (fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, sterols, prenols, polyketides) defined by the LIPID MAPS consortium. PMID:26090714

  5. Chromatography.

    ERIC Educational Resources Information Center

    Brantley, L. Reed, Sr.; Demanche, Edna L.; Klemm, E. Barbara; Kyselka, Will; Phillips, Edwin A.; Pottenger, Francis M.; Yamamoto, Karen N.; Young, Donald B.

    This booklet presents some activities on chromatography. Directions for preparing leaf pigment extracts using alcohol are given, and paper chromatography and thin-layer chromatography are described as modifications of the basic principles of chromatography. (KHR)

  6. A uniform method for the simultaneous blood group phenotyping of Fya , Fyb , Jka , Jkb , S, s̅, P1, k applying lateral-flow technique.

    PubMed

    Caesar, A; Meyer, S; Trost, N; Neuenschwander, K; Geisen, C; Frey, B M; Gassner, C; Schwind, P

    2018-02-01

    A lateral flow assay for simultaneous blood group typing of ABO, RhD, C, E, c, e, Cw and K with stable end-point and without centrifugation is in routine use since several years (MDmulticard ® ). The typing of extended phenotype parameters belonging to the Duffy, Kidd, MNSs blood group systems and others, however, has not yet been demonstrated for this technique. Reliable detection of Fy x , a weak Fy b phenotype with a pronounced quantitative reduction of the number of Fy b antigens on the erythrocyte surface, remains a weakness of current serological blood grouping techniques. The performance characteristics of the following reagents were evaluated in donor and patient samples in lateral flow technology (MDmulticard ® ): Anti-Fy a , -Fy b , -Jk a , -Jk b , -S, -s̅, -P1 and -k. The sensitivity to detect Fy x was in addition evaluated with Fy x positive samples, which had been preselected by MALDI-TOF MS-based genotyping. All results obtained with the MDmulticard ® were in full accordance with those of the CE-certified reference products for all the eight reagent formulations used: Anti-Fy a , -Fy b , -Jk a , -Jk b , -S, -s̅, -P1 and -k. Also, all Fy x phenotypes of the selected population of 93 positive samples, originally identified by MALDI-TOF MS-based genotyping, were reliably detected by the lateral flow assay. Extended phenotype blood group parameters, including the serologically challenging Fy x phenotype, can be determined simultaneously, rapidly and accurately using the lateral flow (MDmulticard ® ) technology, even in cases when IgG class antibodies are the only source of diagnostic antibodies. © 2017 International Society of Blood Transfusion.

  7. Smartphone-Based Dual-Modality Imaging System for Quantitative Detection of Color or Fluorescent Lateral Flow Immunochromatographic Strips

    NASA Astrophysics Data System (ADS)

    Hou, Yafei; Wang, Kan; Xiao, Kun; Qin, Weijian; Lu, Wenting; Tao, Wei; Cui, Daxiang

    2017-04-01

    Nowadays, lateral flow immunochromatographic assays are increasingly popular as a diagnostic tool for point-of-care (POC) test based on their simplicity, specificity, and sensitivity. Hence, quantitative detection and pluralistic popular application are urgently needed in medical examination. In this study, a smartphone-based dual-modality imaging system was developed for quantitative detection of color or fluorescent lateral flow test strips, which can be operated anywhere at any time. In this system, the white and ultra-violet (UV) light of optical device was designed, which was tunable with different strips, and the Sobel operator algorithm was used in the software, which could enhance the identification ability to recognize the test area from the background boundary information. Moreover, this technology based on extraction of the components from RGB format (red, green, and blue) of color strips or only red format of the fluorescent strips can obviously improve the high-signal intensity and sensitivity. Fifty samples were used to evaluate the accuracy of this system, and the ideal detection limit was calculated separately from detection of human chorionic gonadotropin (HCG) and carcinoembryonic antigen (CEA). The results indicated that smartphone-controlled dual-modality imaging system could provide various POC diagnoses, which becomes a potential technology for developing the next-generation of portable system in the near future.

  8. Multiplexed Lateral Flow Test for Detection and Differentiation of Cronobacter sakazakii Serotypes O1 and O2

    PubMed Central

    Scharinger, Eva J.; Dietrich, Richard; Wittwer, Tobias; Märtlbauer, Erwin; Schauer, Kristina

    2017-01-01

    The ubiquitous and opportunistic pathogen Cronobacter sakazakii is responsible for severe meningitis, sepsis, and necrotizing enterocolitis in neonates and infants associated with ingestion of contaminated powdered infant formula (PIF). The current ISO method for isolation and detection of Cronobacter spp. is laborious, time-consuming and expensive. In this study, a multiplexed lateral flow test strip was developed to rapidly detect and simultaneously serotype O1 and O2 C. sakazakii serotypes. The assay is based on two monoclonal antibodies (MAb) that specifically bind to the lipopolysaccharides (LPS) of these pathogens. The test strip provides results very quickly; C. sakazakii could be detected in pure culture within 15 min with a sensitivity of 107 CFU/ml. After non-selective enrichment for 18 h as low as one Cronobacter cell per g PIF could be detected. Moreover, the established lateral flow assay (LFA) offers excellent specificity showing no cross-reactivity with other C. sakazakii serotypes, Cronobacter species or Enterobacteriaceae tested. These characteristics, together with several advantages such as speed, simplicity in performance, low analysis cost, and no requirement of specialized skills or sophisticated equipment make the developed multiplexed LFA suitable for reliable detection and serotyping of C. sakazakii serotypes O1 and O2. PMID:28979257

  9. Multiplexed Lateral Flow Test for Detection and Differentiation of Cronobacter sakazakii Serotypes O1 and O2.

    PubMed

    Scharinger, Eva J; Dietrich, Richard; Wittwer, Tobias; Märtlbauer, Erwin; Schauer, Kristina

    2017-01-01

    The ubiquitous and opportunistic pathogen Cronobacter sakazakii is responsible for severe meningitis, sepsis, and necrotizing enterocolitis in neonates and infants associated with ingestion of contaminated powdered infant formula (PIF). The current ISO method for isolation and detection of Cronobacter spp. is laborious, time-consuming and expensive. In this study, a multiplexed lateral flow test strip was developed to rapidly detect and simultaneously serotype O1 and O2 C. sakazakii serotypes. The assay is based on two monoclonal antibodies (MAb) that specifically bind to the lipopolysaccharides (LPS) of these pathogens. The test strip provides results very quickly; C. sakazakii could be detected in pure culture within 15 min with a sensitivity of 10 7 CFU/ml. After non-selective enrichment for 18 h as low as one Cronobacter cell per g PIF could be detected. Moreover, the established lateral flow assay (LFA) offers excellent specificity showing no cross-reactivity with other C. sakazakii serotypes, Cronobacter species or Enterobacteriaceae tested. These characteristics, together with several advantages such as speed, simplicity in performance, low analysis cost, and no requirement of specialized skills or sophisticated equipment make the developed multiplexed LFA suitable for reliable detection and serotyping of C. sakazakii serotypes O1 and O2.

  10. Triticonazole enantiomers: Separation by supercritical fluid chromatography and the effect of the chromatographic conditions.

    PubMed

    He, Jianfeng; Fan, Jun; Yan, Yilun; Chen, Xiaodong; Wang, Tai; Zhang, Yaomou; Zhang, Weiguang

    2016-11-01

    Enantiomeric pairs of triticonazole have been successfully separated by supercritical fluid chromatography coupled with a tris(3,5-dimethylphenylcarbamoyl) cellulose-coated chiral stationary phase in this work. The effects of co-solvent, dissolution solvent, flow rate, backpressure, and column temperature have been studied in detail with respect to retention, selectivity, and resolution of triticonazole. As indicated, the co-solvents mostly affected the retention factors and resolution, due to the different molecular structure and polarity. In addition, the dissolution solvents, namely, chloromethanes and alcohols, have been also important for enantioseparation because of the different interaction with stationary phase. Higher flow rate and backpressure led to faster elution of the triticonazole molecules, and the change of column temperature showed slight effect on the resolution of triticonazole racemate. Moreover, a comparative separation experiment between supercritical fluid chromatography and high performance liquid chromatography revealed that chiral supercritical fluid chromatography gave the 3.5 times value of R s /t R2 than high performance liquid chromatography, which demonstrated that supercritical fluid chromatography had much higher separation efficiency. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Estimation of the quantification uncertainty from flow injection and liquid chromatography transient signals in inductively coupled plasma mass spectrometry

    NASA Astrophysics Data System (ADS)

    Laborda, Francisco; Medrano, Jesús; Castillo, Juan R.

    2004-06-01

    The quality of the quantitative results obtained from transient signals in high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICPMS) and flow injection-inductively coupled plasma mass spectrometry (FI-ICPMS) was investigated under multielement conditions. Quantification methods were based on multiple-point calibration by simple and weighted linear regression, and double-point calibration (measurement of the baseline and one standard). An uncertainty model, which includes the main sources of uncertainty from FI-ICPMS and HPLC-ICPMS (signal measurement, sample flow rate and injection volume), was developed to estimate peak area uncertainties and statistical weights used in weighted linear regression. The behaviour of the ICPMS instrument was characterized in order to be considered in the model, concluding that the instrument works as a concentration detector when it is used to monitorize transient signals from flow injection or chromatographic separations. Proper quantification by the three calibration methods was achieved when compared to reference materials, although the double-point calibration allowed to obtain results of the same quality as the multiple-point calibration, shortening the calibration time. Relative expanded uncertainties ranged from 10-20% for concentrations around the LOQ to 5% for concentrations higher than 100 times the LOQ.

  12. Electrochemical Detectors in HPLC and Ion Chromatography.

    PubMed

    Horvai, George; Pungor, ErnÕ

    1989-01-01

    Back in 1952, the renowned Polish electrochemist Wiktor Kemula introduced chromato-polarography, 1 i.e., polaro-graphic detection for liquid chromatography. This technique continued to develop slowly until the early 1970s (for a review see Reference 2) when modem high-performance liquid chromatography (HPLC) emerged. This new, highly efficient chromatographc method could only be. used with detectors ensuring low dispersion. It was not easy to modify the dropping mercury electrode cells to satisfy this requirement. However, at the same time, electroanalytical chemists, who already had much experience in using carbon-based electrodes for oxidative detection in flow analysis, put forward the idea of oxidative amperometric detection in liquid chromatography. 3,4 In this technique, solid or quasi-solid (paste) electrodes were used and this made possible the construction of miniaturized cells with just a few microliter volume.

  13. Comprehensive two-dimensional gas chromatography using partial modulation via a pulsed flow valve with a short modulation period.

    PubMed

    Freye, Chris E; Bahaghighat, H Daniel; Synovec, Robert E

    2018-01-15

    Partial modulation via a pulsed flow valve for comprehensive two-dimensional (2D) gas chromatography (GC × GC) is demonstrated, producing narrow peak widths, 2 W b , on the secondary separation dimension, 2 D, coupled with short modulation periods, P M , thus producing a high peak capacity on the 2 D dimension, 2 n c . The GC × GC modulator is a pulse flow valve that injects a pulse of carrier gas at the specified P M , at the connection between the primary, 1 D, column and the 2 D column. Using a commercially available pulse flow valve, this injection technique performs a combination of vacancy chromatography and frontal analysis, whereby each pulse disturbance in the analyte concentration profile as it exits the 1 D column results in data that is readily converted into a 2 D separation. A three-step process converts the raw data into a format analogous to a GC × GC separation, incorporating signal differentiation, baseline correction and conversion to a GC × GC chromatogram representation. A 115-component test mixture with a wide range of boiling points (36-372°C) of nine compound classes is demonstrated using modulation periods of P M = 50, 100, 250, and 500ms, respectively. For the test mixture with a P M of 250ms, peak shapes on 2 D are symmetric with apparent 2 W b ranging from 12 to 45ms producing a 2 n c of ~ 10. Based on the average peak width of 0.93s on the 1 D separation for a time window of 400s, the 1 D peak capacity is 1 n c ∼ 430. Thus, the ideal 2D peak capacity n c,2D is 4300 or a peak capacity production of 650 peaks/min using the P M of 250ms. Additionally, for a P M of 50, 100 and 500ms, the 2 n c are 4, 7, and 12, respectively. Retention times on 2 D, 2 t R , are reproducible having standard deviations less than 1ms. Finally, the processed data is shown to be quantitative, with an average RSD of 4.7% for test analytes. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Online spectrophotometric determination of Fe(II) and Fe(III) by flow injection combined with low pressure ion chromatography

    NASA Astrophysics Data System (ADS)

    Chen, Shujuan; Li, Nan; Zhang, Xinshen; Yang, Dongjing; Jiang, Heimei

    2015-03-01

    A simple and new low pressure ion chromatography combined with flow injection spectrophotometric procedure for determining Fe(II) and Fe(III) was established. It is based on the selective adsorption of low pressure ion chromatography column to Fe(II) and Fe(III), the online reduction reaction of Fe(III) and the reaction of Fe(II) in sodium acetate with phenanthroline, resulting in an intense orange complex with a suitable absorption at 515 nm. Various chemical (such as the concentration of colour reagent, eluant and reductive agent) and instrumental parameters (reaction coil length, reductive coil length and wavelength) were studied and were optimized. Under the optimum conditions calibration graph of Fe(II)/Fe(III) was linear in the Fe(II)/Fe(III) range of 0.040-1.0 mg/L. The detection limit of Fe(III) and Fe(II) was respectively 3.09 and 1.55 μg/L, the relative standard deviation (n = 10) of Fe(II) and Fe(III) 1.89% and 1.90% for 0.5 mg/L of Fe(II) and Fe(III) respectively. About 2.5 samples in 1 h can be analyzed. The interfering effects of various chemical species were studied. The method was successfully applied in the determination of water samples.

  15. A free-flowing soap film combined with cavity ring-down spectroscopy as a detection system for liquid chromatography.

    PubMed

    Vogelsang, Markus; Welsch, Thomas; Jones, Harold

    2010-05-07

    We have shown that a free-flowing soap film has sufficiently high-quality optical properties to allow it to be used in the cavity of a ring-down spectrometer (CRDS). The flow rates required to maintain a stable soap film were similar to those used in liquid chromatography and thus allowed interfacing with an HPLC system for use as an optical detector. We have investigated the properties of the system in a relevant analytical application. The soap film/CRDS combination was used at 355 nm as a detector for the separation of a mixture of nitroarenes. These compounds play a role in the residue analysis of areas contaminated with explosives and their decomposition products. In spite of the short absorption path length (9 microm) obtained by the soap film, the high-sensitivity of CRDS allowed a limit of detection of 4 x 10(-6) in absorption units (AU) or less than 17 fmol in the detection volume to be achieved. Copyright (c) 2009 Elsevier B.V. All rights reserved.

  16. Hydrodynamic chromatography of macromolecules using polymer monolithic columns.

    PubMed

    Edam, Rob; Eeltink, Sebastiaan; Vanhoutte, Dominique J D; Kok, Wim Th; Schoenmakers, Peter J

    2011-12-02

    The selectivity window of size-based separations of macromolecules was tailored by tuning the macropore size of polymer monolithic columns. Monolithic materials with pore sizes ranging between 75 nm and 1.2 μm were prepared in situ in large I.D. columns. The dominant separation mechanism was hydrodynamic chromatography in the flow-through pores. The calibration curves for synthetic polymers matched with the elution behavior by HDC separations in packed columns with 'analyte-to-pore' aspect ratios (λ) up to 0.2. For large-macropore monoliths, a deviation in retention behavior was observed for small polystyrene polymers (M(r)<20 kDa), which may be explained by a combined HDC-SEC mechanism for λ<0.02. The availability of monoliths with very narrow pore sizes allowed investigation of separations at high λ values. For high-molecular weight polymers (M(r)>300,000 Da) confined in narrow channels, the separation strongly depended on flow rate. Flow-rate dependent elution behavior was evaluated by calculation of Deborah numbers and confirmed to be outside the scope of classic shear deformation or slalom chromatography. Shear-induced forces acting on the periphery of coiled polymers in solution may be responsible for flow-rate dependent elution. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Liquid-phase chromatography detector

    DOEpatents

    Voigtman, Edward G.; Winefordner, James D.; Jurgensen, Arthur R.

    1983-01-01

    A liquid-phase chromatography detector comprising a flow cell having an inlet tubular conduit for receiving a liquid chromatographic effluent and discharging it as a flowing columnar stream onto a vertically adjustable receiving surface spaced apart from and located vertically below and in close proximity to the discharge end of the tubular conduit; a receiver adapted to receive liquid overflowing from the receiving surface; an exit conduit for continuously removing liquid from the receiver; a light source for focussing fluorescence-producing light pulses on the flowing columnar stream as it passes from the outlet of the conduit to the receiving surface and a fluorescence detector to detect the produced fluorescence; a source of light pulse for producing acoustic waves in the columnar stream as it passes from the conduit outlet to the receiving surface; and a piezoelectric transducer adapted to detect those waves; and a source of bias voltage applied to the inlet tubular conduit and adapted to produce ionization of the liquid flowing through the flow cell so as to produce photocurrents therein and an electrical system to detect and record the photocurrents. This system is useful in separating and detecting individual chemical compounds from mixtures thereof.

  18. Zonal Rate Model for Axial and Radial Flow Membrane Chromatography. Part I: Knowledge Transfer Across Operating Conditions and Scales

    PubMed Central

    Ghosh, Pranay; Vahedipour, Kaveh; Lin, Min; Vogel, Jens H; Haynes, Charles A; von Lieres, Eric

    2013-01-01

    The zonal rate model (ZRM) has previously been applied for analyzing the performance of axial flow membrane chromatography capsules by independently determining the impacts of flow and binding related non-idealities on measured breakthrough curves. In the present study, the ZRM is extended to radial flow configurations, which are commonly used at larger scales. The axial flow XT5 capsule and the radial flow XT140 capsule from Pall are rigorously analyzed under binding and non-binding conditions with bovine serum albumin (BSA) as test molecule. The binding data of this molecule is much better reproduced by the spreading model, which hypothesizes different binding orientations, than by the well-known Langmuir model. Moreover, a revised cleaning protocol with NaCl instead of NaOH and minimizing the storage time has been identified as most critical for quantitatively reproducing the measured breakthrough curves. The internal geometry of both capsules is visualized by magnetic resonance imaging (MRI). The flow in the external hold-up volumes of the XT140 capsule was found to be more homogeneous as in the previously studied XT5 capsule. An attempt for model-based scale-up was apparently impeded by irregular pleat structures in the used XT140 capsule, which might lead to local variations in the linear velocity through the membrane stack. However, the presented approach is universal and can be applied to different capsules. The ZRM is shown to potentially help save valuable material and time, as the experiments required for model calibration are much cheaper than the predicted large-scale experiment at binding conditions. Biotechnol. Bioeng. 2013; 110: 1129–1141. © 2012 Wiley Periodicals, Inc. PMID:23097218

  19. Development of an immunochromatographic lateral flow device for rapid diagnosis of Vibrio cholerae O1 serotype Ogawa.

    PubMed

    Chen, Weixian; Zhang, Jun; Lu, Gang; Yuan, Zuowei; Wu, Qian; Li, Jingjing; Xu, Guiping; He, An; Zheng, Jian; Zhang, Juan

    2014-04-01

    Cholera is an acute malignant infectious disease caused by the bacteria Vibrio cholerae leading to severe dehydrating diarrhea and vomiting, even high rates of mortality in some cases. However, the prevention of the epidemic disease is achievable if proper sanitation practices are followed, provided the accurate and prompt diagnosis of each prevalent serotype in cholera epidemic. The current gold standard of bacterial culture is inadequate for rapid diagnosis. Our aim is to develop an immunochromatographic test format for O1 serotype Ogawa diagnosis and provide the need for better epidemic prevention and early response. The monoclonal antibodies were raised in conventional method and subsequently screened for a match pair. A variety of related and unrelated bacteria strains recruited were employed to test their sensitivity, specificity etc. by indirect ELISA. The human fecal samples were used to test the final lateral-flow device product to satisfy the measurement requirement. A new monoclonal antibody (McAb) pair, named IXiao₃G₆ and IXiao₁D₉, was generated, which is specifically against V. cholerae O1 serotype Ogawa. Additionally, we developed an immunochromatographic lateral flow device (LFD) using this McAb pair for the highly specific and rapid (5 min) detection of Ogawa. Our product has advantages of simplicity and precision, and can benefit the scene and elementary medical institutions. Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  20. Micro-polarimeter for high performance liquid chromatography

    DOEpatents

    Yeung, Edward E.; Steenhoek, Larry E.; Woodruff, Steven D.; Kuo, Jeng-Chung

    1985-01-01

    A micro-polarimeter interfaced with a system for high performance liquid chromatography, for quantitatively analyzing micro and trace amounts of optically active organic molecules, particularly carbohydrates. A flow cell with a narrow bore is connected to a high performance liquid chromatography system. Thin, low birefringence cell windows cover opposite ends of the bore. A focused and polarized laser beam is directed along the longitudinal axis of the bore as an eluent containing the organic molecules is pumped through the cell. The beam is modulated by air gap Faraday rotators for phase sensitive detection to enhance the signal to noise ratio. An analyzer records the beams's direction of polarization after it passes through the cell. Calibration of the liquid chromatography system allows determination of the quantity of organic molecules present from a determination of the degree to which the polarized beam is rotated when it passes through the eluent.

  1. A CCD-based reader combined with CdS quantum dot-labeled lateral flow strips for ultrasensitive quantitative detection of CagA

    NASA Astrophysics Data System (ADS)

    Gui, Chen; Wang, Kan; Li, Chao; Dai, Xuan; Cui, Daxiang

    2014-02-01

    Immunochromatographic assays are widely used to detect many analytes. CagA is proved to be associated closely with initiation of gastric carcinoma. Here, we reported that a charge-coupled device (CCD)-based test strip reader combined with CdS quantum dot-labeled lateral flow strips for quantitative detection of CagA was developed, which used 365-nm ultraviolet LED as the excitation light source, and captured the test strip images through an acquisition module. Then, the captured image was transferred to the computer and was processed by a software system. A revised weighted threshold histogram equalization (WTHE) image processing algorithm was applied to analyze the result. CdS quantum dot-labeled lateral flow strips for detection of CagA were prepared. One hundred sera samples from clinical patients with gastric cancer and healthy people were prepared for detection, which demonstrated that the device could realize rapid, stable, and point-of-care detection, with a sensitivity of 20 pg/mL.

  2. Enhancement of lateral flow assay performance by electromagnetic relocation of reporter particles

    PubMed Central

    Jacinto, Maria João; Trabuco, João R. C.; Vu, Binh V.; Garvey, Gavin; Khodadady, Mohammad; Azevedo, Ana M.; Aires-Barros, Maria Raquel; Chang, Long; Kourentzi, Katerina; Litvinov, Dmitri

    2018-01-01

    Lateral flow assays (LFAs) are a widely-used point-of care diagnostic format, but suffer from limited analytical sensitivity, especially when read by eye. It has recently been reported that LFA performance can be improved by using magnetic reporter particles and an external magnetic field applied at the test line. The mechanism of sensitivity/performance enhancement was suggested to be concentration/retardation of reporter particles at the test line. Here we demonstrate an additional mechanism of particle relocation where reporter particles from the lower depths of the translucent LFA strip relocate to more-visible locations nearer to the top surface, producing a more visible signal. With a magnetic field we observed an improvement in sensitivity of human chorionic gonadotropin (hCG) detection from 1.25 ng/mL to 0.31 ng/mL. We also observed an increase of the color intensity per particle in test lines when the magnetic field was present. PMID:29309424

  3. Effects of Soluble Surfactant on Lateral Migration of a Bubble in a Shear Flow

    NASA Astrophysics Data System (ADS)

    Muradoglu, Metin; Tryggvason, Gretar

    2014-11-01

    Motivated by the recent experimental study of Takagi et al. (2008), direct numerical simulations are performed to examine effects of soluble surfactant on the lateral migration of a deformable bubble in a pressure-driven channel flow. The interfacial and bulk surfactant concentration evolution equations are solved fully coupled with the incompressible Navier-Stokes equations. A non-linear equation of state is used to relate interfacial surface tension to surfactant concentration at the interface. A multiscale method is developed to handle the mass exchange between the interface and bulk fluid at high Peclet numbers, using a boundary-layer approximation next to the bubble and a relatively coarse grid for the rest of the flow. It is found that the surfactant induced Marangoni stresses can dominate over the shear-induced lift force and thus alter the behavior of the bubble completely, i.e., the contaminated bubble drifts away from the channel wall and stabilizes at the center of the channel in contrast with the corresponding clean bubble that drifts toward the wall and stabilizes near the wall. The Scientific and Technical Research Council of Turkey (TUBITAK), Grant 112M181 and Turkish Academy of Sciences (TUBA).

  4. A lateral flow assay (LFA) for the rapid detection of extraparenchymal neurocysticercosis using cerebrospinal fluid.

    PubMed

    Fleury, Agnes; Sastre, Patricia; Sciutto, Edda; Correia, Silvia; Monedero, Alejandro; Toledo, Andrea; Hernandez, Maricela; Harrison, Leslie J S; Parkhouse, R Michael E

    2016-10-27

    A lateral flow assay (LFA) for the diagnosis and monitoring of extraparenchymal neurocysticercosis, has been developed. The assay is based on the use of the monoclonal antibody HP10, and when applied to cerebrospinal fluid, correctly identified 34 cases of active extraparenchymal neurocysticercosis, but was negative with 26 samples from treated and cured neurocysticercosis patients and with 20 samples from unrelated neurological diseases. There was complete agreement between the HP10 Ag-ELISA results and the HP10-LFA. The HP10-LFA thus has utility for diagnosis and treatment of extraparenchymal neurocysticercosis, frequently a more dangerous form of the infection. Copyright © 2016. Published by Elsevier Inc.

  5. Characterization of vitamin D3 metabolites using continuous-flow fast atom bombardment tandem mass spectrometry and high-performance liquid chromatography.

    PubMed

    Yeung, B; Vouros, P; Reddy, G S

    1993-08-13

    A mass spectrometric method for the detection of vitamin D3 metabolites is described. This method involves the derivatization of the metabolites by cycloaddition with 4-phenyl-1,2,4-triazoline-3,5-dione, followed by their characterization by continuous-flow fast atom bombardment (CF-FAB) tandem mass spectrometry (MS-MS) and high-performance liquid chromatography (HPLC). Using HPLC, this derivatization has been shown to increase the UV detectability of 25-hydroxyvitamin D3 by about 5-fold. The FAB spectra of the adducts are dominated by peaks corresponding to a protonated molecule and a fragment ion derived in part from the loss of the side chain. Multiple reaction monitoring (MRM) of this transition by MS-MS may be utilized for trace level analysis of vitamin D metabolites. Sample introduction by flow injection yields detection limits in the low nanogram to high picogram range, whereas the use of on-line capillary LC has been found to decrease the detection limits to the low picogram level.

  6. Development of Lateral Flow Immunoassay for Antigen Detection in Human Angiostrongylus cantonensis Infection

    PubMed Central

    Chen, Mu-Xin; Chen, Jia-Xu; Chen, Shao-Hong; Huang, Da-Na; Ai, Lin; Zhang, Ren-Li

    2016-01-01

    Angiostrongyliasis is difficult to be diagnosed for the reason that no ideal method can be used. Serologic tests require specific equipment and are not always available in poverty-stricken zone and are time-consuming. A lateral flow immunoassay (LFIA) may be useful for angiostrongyliasis control. We established a LFIA for the diagnosis of angiostrongyliasis based on 2 monoclonal antibodies (mAbs) against antigens of Angiostrongylus cantonensis adults. The sensitivity and specificity were 91.1% and 100% in LFIA, while those of commercial ELISA kit was 97.8% and 86.3%, respectively. Youden index was 0.91 in LFIA and 0.84 in commercial ELISA kit. LFIA showed detection limit of 1 ng/ml of A. cantonensis ES antigens. This LFIA was simple, rapid, highly sensitive and specific, which opened an alternative approach for the diagnosis of human angiostrongyliasis. PMID:27417097

  7. Broken flow symmetry explains the dynamics of small particles in deterministic lateral displacement arrays

    PubMed Central

    Kim, Sung-Cheol; Wunsch, Benjamin H.; Hu, Huan; Smith, Joshua T.; Stolovitzky, Gustavo

    2017-01-01

    Deterministic lateral displacement (DLD) is a technique for size fractionation of particles in continuous flow that has shown great potential for biological applications. Several theoretical models have been proposed, but experimental evidence has demonstrated that a rich class of intermediate migration behavior exists, which is not predicted. We present a unified theoretical framework to infer the path of particles in the whole array on the basis of trajectories in a unit cell. This framework explains many of the unexpected particle trajectories reported and can be used to design arrays for even nanoscale particle fractionation. We performed experiments that verify these predictions and used our model to develop a condenser array that achieves full particle separation with a single fluidic input. PMID:28607075

  8. Broken flow symmetry explains the dynamics of small particles in deterministic lateral displacement arrays.

    PubMed

    Kim, Sung-Cheol; Wunsch, Benjamin H; Hu, Huan; Smith, Joshua T; Austin, Robert H; Stolovitzky, Gustavo

    2017-06-27

    Deterministic lateral displacement (DLD) is a technique for size fractionation of particles in continuous flow that has shown great potential for biological applications. Several theoretical models have been proposed, but experimental evidence has demonstrated that a rich class of intermediate migration behavior exists, which is not predicted. We present a unified theoretical framework to infer the path of particles in the whole array on the basis of trajectories in a unit cell. This framework explains many of the unexpected particle trajectories reported and can be used to design arrays for even nanoscale particle fractionation. We performed experiments that verify these predictions and used our model to develop a condenser array that achieves full particle separation with a single fluidic input.

  9. Contribution of lateral terrestrial water flows to the regional hydrological cycle: A joint soil-atmospheric moisture tagging procedure with WRF-Hydro

    NASA Astrophysics Data System (ADS)

    Arnault, Joel; Wei, Jianhui; Zhang, Zhenyu; Wagner, Sven; Kunstmann, Harald

    2017-04-01

    Water resources management requires an accurate knowledge of the behavior of the regional hydrological cycle components, including precipitation, evapotranspiration, river discharge and soil water storage. Atmospheric models such as the Weather Research and Forecasting (WRF) model provide a tool to evaluate these components. The main drawback of these atmospheric models, however, is that the terrestrial segment of the hydrological cycle is reduced to vertical infiltration, and that lateral terrestrial water flows are neglected. Recent model developments have focused on coupled atmospheric-hydrological modeling systems, such as WRF-hydro, in order to take into account subsurface, overland and river flow. The aim of this study is to investigate the contribution of lateral terrestrial water flows to the regional hydrological cycle, with the help of a joint soil-atmospheric moisture tagging procedure. This procedure is the extended version of an existing atmospheric moisture tagging method developed in WRF and WRF-Hydro (Arnault et al. 2017). It is used to quantify the partitioning of precipitation into water stored in the soil, runoff, evapotranspiration, and potentially subsequent precipitation through regional recycling. An application to a high precipitation event on 23 June 2009 in the upper Danube river basin, Germany and Austria, is presented. Precipitating water during this day is tagged for the period 2009-2011. Its contribution to runoff and evapotranspiration decreases with time, but is still not negligible in the summer 2011. At the end of the study period, less than 5 % of the precipitating water on 23 June 2009 remains in the soil. The additionally resolved lateral terrestrial water flows in WRF-Hydro modify the partitioning between surface and underground runoff, in association with a slight increase of evapotranspiration and recycled precipitation. Reference: Arnault, J., R. Knoche, J. Wei, and H. Kunstmann (2016), Evaporation tagging and atmospheric

  10. Paper-based enzymatic microfluidic fuel cell: From a two-stream flow device to a single-stream lateral flow strip

    NASA Astrophysics Data System (ADS)

    González-Guerrero, Maria José; del Campo, F. Javier; Esquivel, Juan Pablo; Giroud, Fabien; Minteer, Shelley D.; Sabaté, Neus

    2016-09-01

    This work presents a first approach towards the development of a cost-effective enzymatic paper-based glucose/O2 microfluidic fuel cell in which fluid transport is based on capillary action. A first fuel cell configuration consists of a Y-shaped paper device with the fuel and the oxidant flowing in parallel over carbon paper electrodes modified with bioelectrocatalytic enzymes. The anode consists of a ferrocenium-based polyethyleneimine polymer linked to glucose oxidase (GOx/Fc-C6-LPEI), while the cathode contains a mixture of laccase, anthracene-modified multiwall carbon nanotubes, and tetrabutylammonium bromide-modified Nafion (MWCNTs/laccase/TBAB-Nafion). Subsequently, the Y-shaped configuration is improved to use a single solution containing both, the anolyte and the catholyte. Thus, the electrolytes pHs of the fuel and the oxidant solutions are adapted to an intermediate pH of 5.5. Finally, the fuel cell is run with this single solution obtaining a maximum open circuit of 0.55 ± 0.04 V and a maximum current and power density of 225 ± 17 μA cm-2 and 24 ± 5 μW cm-2, respectively. Hence, a power source closer to a commercial application (similar to conventional lateral flow test strips) is developed and successfully operated. This system can be used to supply the energy required to power microelectronics demanding low power consumption.

  11. Application of a chromatography model with linear gradient elution experimental data to the rapid scale-up in ion-exchange process chromatography of proteins.

    PubMed

    Ishihara, Takashi; Kadoya, Toshihiko; Yamamoto, Shuichi

    2007-08-24

    We applied the model described in our previous paper to the rapid scale-up in the ion exchange chromatography of proteins, in which linear flow velocity, column length and gradient slope were changed. We carried out linear gradient elution experiments, and obtained data for the peak salt concentration and peak width. From these data, the plate height (HETP) was calculated as a function of the mobile phase velocity and iso-resolution curve (the separation time and elution volume relationship for the same resolution) was calculated. The scale-up chromatography conditions were determined by the iso-resolution curve. The scale-up of the linear gradient elution from 5 to 100mL and 2.5L column sizes was performed both by the separation of beta-lactoglobulin A and beta-lactoglobulin B with anion-exchange chromatography and by the purification of a recombinant protein with cation-exchange chromatography. Resolution, recovery and purity were examined in order to verify the proposed method.

  12. Mobile Phone Ratiometric Imaging Enables Highly Sensitive Fluorescence Lateral Flow Immunoassays without External Optical Filters.

    PubMed

    Shah, Kamal G; Singh, Vidhi; Kauffman, Peter C; Abe, Koji; Yager, Paul

    2018-05-14

    Paper-based diagnostic tests based on the lateral flow immunoassay concept promise low-cost, point-of-care detection of infectious diseases, but such assays suffer from poor limits of detection. One factor that contributes to poor analytical performance is a reliance on low-contrast chromophoric optical labels such as gold nanoparticles. Previous attempts to improve the sensitivity of paper-based diagnostics include replacing chromophoric labels with enzymes, fluorophores, or phosphors at the expense of increased fluidic complexity or the need for device readers with costly optoelectronics. Several groups, including our own, have proposed mobile phones as suitable point-of-care readers due to their low cost, ease of use, and ubiquity. However, extant mobile phone fluorescence readers require costly optical filters and were typically validated with only one camera sensor module, which is inappropriate for potential point-of-care use. In response, we propose to couple low-cost ultraviolet light-emitting diodes with long Stokes-shift quantum dots to enable ratiometric mobile phone fluorescence measurements without optical filters. Ratiometric imaging with unmodified smartphone cameras improves the contrast and attenuates the impact of excitation intensity variability by 15×. Practical application was shown with a lateral flow immunoassay for influenza A with nucleoproteins spiked into simulated nasal matrix. Limits of detection of 1.5 and 2.6 fmol were attained on two mobile phones, which are comparable to a gel imager (1.9 fmol), 10× better than imaging gold nanoparticles on a scanner (18 fmol), and >2 orders of magnitude better than gold nanoparticle-labeled assays imaged with mobile phones. Use of the proposed filter-free mobile phone imaging scheme is a first step toward enabling a new generation of highly sensitive, point-of-care fluorescence assays.

  13. A Lateral Flow Rapid Test for Human Toxocariasis Developed Using Three Toxocara canis Recombinant Antigens.

    PubMed

    Yunus, Muhammad Hafiznur; Tan Farrizam, Siti Naqiuyah; Abdul Karim, Izzati Zahidah; Noordin, Rahmah

    2018-01-01

    Laboratory diagnosis of toxocariasis is still a challenge especially in developing endemic countries with polyparasitism. In this study, three Toxocara canis recombinant antigens, rTES-26, rTES-30, and rTES-120, were expressed and used to prepare lateral flow immunoglobulin G4 (IgG4) dipsticks. The concordance of the results of the rapid test (comprising three dipsticks) with a commercial IgG-enzyme-linked immunosorbent assay (ELISA) (Cypress Diagnostics, Belgium) was compared against the concordance of two other commercial IgG-ELISA kits (Bordier, Switzerland and NovaTec, Germany) with the Cypress kit. Using Toxocara- positive samples, the concordance of the dipstick dotted with rTES-26, rTES-30, and rTES-120 was 41.4% (12/29), 51.7% (15/29), and 72.4% (21/29), respectively. When positivity with any dipstick was considered as an overall positive rapid test result, the concordance with the Cypress kit was 93% (27/29). Meanwhile, when compared with the results of the Cypress kit, the concordance of IgG-ELISA from NovaTec and Bordier was 100% (29/29) and 89.7% (26/29), respectively. Specific IgG4 has been recognized as a marker of active infection for several helminthic diseases; therefore, the two non-concordant results of the rapid test when compared with the NovaTec IgG-ELISA kit may be from samples of people with non-active infection. All the three dipsticks showed 100% (50/50) concordance with the Cypress kit when tested with serum from individuals who were healthy and with other infections. In conclusion, the lateral flow rapid test is potentially a good, fast, and easy test for toxocariasis. Next, further validation studies and development of a test with the three antigens in one dipstick will be performed.

  14. Simple System for Isothermal DNA Amplification Coupled to Lateral Flow Detection

    PubMed Central

    Roskos, Kristina; Hickerson, Anna I.; Lu, Hsiang-Wei; Ferguson, Tanya M.; Shinde, Deepali N.; Klaue, Yvonne; Niemz, Angelika

    2013-01-01

    Infectious disease diagnosis in point-of-care settings can be greatly improved through integrated, automated nucleic acid testing devices. We have developed an early prototype for a low-cost system which executes isothermal DNA amplification coupled to nucleic acid lateral flow (NALF) detection in a mesofluidic cartridge attached to a portable instrument. Fluid handling inside the cartridge is facilitated through one-way passive valves, flexible pouches, and electrolysis-driven pumps, which promotes a compact and inexpensive instrument design. The closed-system disposable prevents workspace amplicon contamination. The cartridge design is based on standard scalable manufacturing techniques such as injection molding. Nucleic acid amplification occurs in a two-layer pouch that enables efficient heat transfer. We have demonstrated as proof of principle the amplification and detection of Mycobacterium tuberculosis (M.tb) genomic DNA in the cartridge, using either Loop Mediated Amplification (LAMP) or the Exponential Amplification Reaction (EXPAR), both coupled to NALF detection. We envision that a refined version of this cartridge, including upstream sample preparation coupled to amplification and detection, will enable fully-automated sample-in to answer-out infectious disease diagnosis in primary care settings of low-resource countries with high disease burden. PMID:23922706

  15. Liquid-phase chromatography detector

    DOEpatents

    Voigtman, E.G.; Winefordner, J.D.; Jurgensen, A.R.

    1983-11-08

    A liquid-phase chromatography detector comprises a flow cell having an inlet tubular conduit for receiving a liquid chromatographic effluent and discharging it as a flowing columnar stream onto a vertically adjustable receiving surface spaced apart from and located vertically below and in close proximity to the discharge end of the tubular conduit; a receiver adapted to receive liquid overflowing from the receiving surface; an exit conduit for continuously removing liquid from the receiver; a light source for focusing fluorescence-producing light pulses on the flowing columnar stream as it passes from the outlet of the conduit to the receiving surface and a fluorescence detector to detect the produced fluorescence; a source of light pulse for producing acoustic waves in the columnar stream as it passes from the conduit outlet to the receiving surface; and a piezoelectric transducer adapted to detect those waves; and a source of bias voltage applied to the inlet tubular conduit and adapted to produce ionization of the liquid flowing through the flow cell so as to produce photocurrents therein and an electrical system to detect and record the photocurrents. This system is useful in separating and detecting individual chemical compounds from mixtures thereof. 5 figs.

  16. Top-down and bottom-up lipidomic analysis of rabbit lipoproteins under different metabolic conditions using flow field-flow fractionation, nanoflow liquid chromatography and mass spectrometry.

    PubMed

    Byeon, Seul Kee; Kim, Jin Yong; Lee, Ju Yong; Chung, Bong Chul; Seo, Hong Seog; Moon, Myeong Hee

    2015-07-31

    This study demonstrated the performances of top-down and bottom-up approaches in lipidomic analysis of lipoproteins from rabbits raised under different metabolic conditions: healthy controls, carrageenan-induced inflammation, dehydration, high cholesterol (HC) diet, and highest cholesterol diet with inflammation (HCI). In the bottom-up approach, the high density lipoproteins (HDL) and the low density lipoproteins (LDL) were size-sorted and collected on a semi-preparative scale using a multiplexed hollow fiber flow field-flow fractionation (MxHF5), followed by nanoflow liquid chromatography-ESI-MS/MS (nLC-ESI-MS/MS) analysis of the lipids extracted from each lipoprotein fraction. In the top-down method, size-fractionated lipoproteins were directly infused to MS for quantitative analysis of targeted lipids using chip-type asymmetrical flow field-flow fractionation-electrospray ionization-tandem mass spectrometry (cAF4-ESI-MS/MS) in selected reaction monitoring (SRM) mode. The comprehensive bottom-up analysis yielded 122 and 104 lipids from HDL and LDL, respectively. Rabbits within the HC and HCI groups had lipid patterns that contrasted most substantially from those of controls, suggesting that HC diet significantly alters the lipid composition of lipoproteins. Among the identified lipids, 20 lipid species that exhibited large differences (>10-fold) were selected as targets for the top-down quantitative analysis in order to compare the results with those from the bottom-up method. Statistical comparison of the results from the two methods revealed that the results were not significantly different for most of the selected species, except for those species with only small differences in concentration between groups. The current study demonstrated that top-down lipid analysis using cAF4-ESI-MS/MS is a powerful high-speed analytical platform for targeted lipidomic analysis that does not require the extraction of lipids from blood samples. Copyright © 2015 Elsevier B

  17. Recombinase Polymerase Amplification Combined with Lateral Flow Strip for Listeria monocytogenes Detection in Food.

    PubMed

    Du, Xin-Jun; Zang, Yu-Xuan; Liu, Hai-Bin; Li, Ping; Wang, Shuo

    2018-04-01

    Listeria monocytogenes is an important food-borne pathogenic bacterium that causes human disease, resulting in economic losses worldwide. The current detection methods for L. monocytogenes are not well suited for direct field testing because they involve complicated, time-consuming operations. A simple, efficient method is vital for L. monocytogenes detection. In this study, we combined isothermal recombinase polymerase amplification (RPA) with a lateral flow (LF) strip to rapidly and reliably detect L. monocytogenes. In the presence of biotin- and digoxin-modified primers, RPA produced numerous digoxin- and biotin-attached duplex DNA products. These products were detected on an LF strip via dual immunoreactions (digoxin on the duplex DNA reacted with the anti-digoxin antibody on the gold nanoparticle (Au-NP) and the biotin on the duplex DNA captured by the streptavidin on the LF test zone). The accumulation of Au-NPs produced characteristic bands, enabling the visual detection of L. monocytogenes without instrumentation. This assay could be used to detect L. monocytogenes within 15 min, including DNA amplification with RPA for 10 min at 39 °C and visualization of the amplicons by LF strips for 5 min. Experiments confirmed a detection limit as low as 300 fg of DNA and 1.5 × 10 1 CFU in pure cultures. Furthermore, RPA-LF exhibited no cross-reactions with pathogens. Evaluation of the method with food samples indicated that the detection limit was substantially improved to 1.5 × 10° CFU for the original bacterial content in 25 g/mL samples after enrichment for 6 hr. RPA-LF can be used as a sensitive and rapid detection technique for L. monocytogenes. Recombinase polymerase amplification (RPA) can amplify target DNA at 37 to 42 °C without a thermal cycler. Lateral flow (LF) strips are portable, cheap and easy to operate. RPA combined with LF strips to detect Listeria monocytogenes can be widely used in remote areas. © 2018 Institute of Food Technologists®.

  18. Quantification of Hydroxychloroquine in Blood Using Turbulent Flow Liquid Chromatography-Tandem Mass Spectrometry (TFLC-MS/MS).

    PubMed

    Chambliss, Allison B; Füzéry, Anna K; Clarke, William A

    2016-01-01

    Hydroxychloroquine (HQ) is used routinely in the treatment of autoimmune disorders such as rheumatoid arthritis and lupus erythematosus. Issues such as marked pharmacokinetic variability and patient non-compliance make therapeutic drug monitoring of HQ a useful tool for management of patients taking this drug. Quantitative measurements of HQ may aid in identifying poor efficacy as well as provide reliable information to distinguish patient non-compliance from refractory disease. We describe a rapid 7-min assay for the accurate and precise measurement of HQ concentrations in 100 μL samples of human blood using turbulent flow liquid chromatography coupled to tandem mass spectrometry. HQ is isolated from EDTA whole blood after a simple extraction with its deuterated analog, hydroxychloroquine-d4, in 0.33 M perchloric acid. Samples are then centrifuged and injected onto the TFLC-MS/MS system. Quantification is performed using a nine-point calibration curve that is linear over a wide range (15.7-4000 ng/mL) with precisions of <5 %.

  19. Validation of a rapid lateral flow test for the simultaneous determination of β-lactam drugs and flunixin in raw milk.

    PubMed

    Douglas, David; Banaszewski, Katie; Juskelis, Rima; Al-Taher, Fadwa; Chen, Yang; Cappozzo, Jack; McRobbie, Lindsay; Salter, Robert S

    2012-07-01

    β-Lactam antibiotics are the most commonly used drugs on dairy farms. β-Lactam residues in milk are kept out of the human milk supply with good agricultural practices and mandatory truck screening performed by the dairy industry under Appendix N of the Pasteurized Milk Ordinance. Flunixin, a nonsteroidal and anti-inflammatory drug, appears in dairy cattle tissue residues with a frequency similar to the occurrence of penicillin G. This creates concern that flunixin residues could be in milk and would go undetected under current milk screening programs. A single test that combines mandatory β-lactam screening with voluntary flunixin screening is an economical approach for monitoring and controlling for potential flunixin or 5-hydroxyflunixin, the primary flunixin metabolite marker in milk. The objective of this study was to validate a β-lactam and flunixin rapid lateral flow test (LFT) and compare the results obtained with a liquid chromatography-triple quadrupole tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of flunixin and 5-hydroxyflunixin in raw milk with a limit of detection of , 1 ppb, equivalent to 1 ng/ml. Using the LFT, three combined manufactured lots of test strips detected penicillin G at 2.0 ppb, ampicillin at 6.8 ppb, amoxicillin at 5.9 ppb, cephapirin at 13.4 ppb, ceftiofur (total metabolites) at 63 ppb, and 5-hydroxyflunixin at 1.9 ppb at least 90% of the time with 95% confidence. The LFT also detected incurred flunixin milk samples that were analyzed with the LC-MS/MS and diluted to tolerance in raw milk. The detection levels for the LFT are lower than the U.S. safe levels or tolerances and qualify the test to be used in compliance with U.S. milk screening programs.

  20. Quantitative Lateral Flow Assays for Salivary Biomarker Assessment: A Review

    PubMed Central

    Miočević, Olga; Cole, Craig R.; Laughlin, Mary J.; Buck, Robert L.; Slowey, Paul D.; Shirtcliff, Elizabeth A.

    2017-01-01

    Saliva is an emerging biofluid with a significant number of applications in use across research and clinical settings. The present paper explores the reasons why saliva has grown in popularity in recent years, balancing both the potential strengths and weaknesses of this biofluid. Focusing on reasons why saliva is different from other common biological fluids such as blood, urine, or tears, we review how saliva is easily obtained, with minimal risk to the donor, and reduced costs for collection, transportation, and analysis. We then move on to a brief review of the history and progress in rapid salivary testing, again reviewing the strengths and weaknesses of rapid immunoassays (e.g., lateral flow immunoassay) compared to more traditional immunoassays. We consider the potential for saliva as an alternative biofluid in a setting where rapid results are important. We focus the review on salivary tests for small molecule biomarkers using cortisol as an example. Such salivary tests can be applied readily in a variety of settings and for specific measurement purposes, providing researchers and clinicians with opportunities to assess biomarkers in real time with lower transportation, collection, and analysis costs, faster turnaround time, and minimal training requirements. We conclude with a note of cautious optimism that the field will soon gain the ability to collect and analyze salivary specimens at any location and return viable results within minutes. PMID:28660183

  1. Development of Lateral Flow Immunochromatographic Strips for Micropollutant Screening Using Colorants of Aptamer-Functionalized Nanogold Particles, Part II: Experimental Verification with Aflatoxin B1 and Chloramphenicol.

    PubMed

    Zhang, Shan; Zhao, Shuai; Wang, Sai; Liu, Jiahui; Dong, Yiyang

    2018-05-09

    Lateral flow immunochromatographic strips based on colorants of aptamer-functionalized nanogold particles weredeveloped for the detection of micropollutants aflatoxin B1 (AFB1) and chloramphenicol (CAP). The lateral flow immunochromatographic strip was based on a competitive reaction of thiolated-aptamer between micropollutants and bio-DNA probe-streptavidin as capture material immobilized at the test line. General crucial parameters that might influence the sensitivity have been systematically investigated. To test the effectiveness and applicability of the optimized conditions, two structurally unrelated micropollutants, that is, AFB1 and CAP, were chosen for detection. In the present study, lateral flow immunochromatographic strips for AFB1 and CAP analysis by combining the high selectivity and affinity of aptamers with the unique optical properties of nanogold in municipal water samples were reported for the first time. With the optimized conditions, the immunochromatographic strip showed a visual LOD of 10 ppb and a quantitative LOD of 1.05 ppb using an immunochromatographic reader for AFB1 detection and a quantitative LOD of 63.4 ppb using an immunochromatographic reader for CAP detection. Furthermore, the sensitive strip provided a good linear detection range of approximately 0-50 ppm for AFB1 detection and a wider liner detection range of approximately 0-160 ppm for CAP detection. Moreover, the immunochromatographic strip provided recovery rates for water samples of 90-110% in the AFB1 analysis and 84-108% in the CAP analysis. The results demonstrated that the immunochromatographic strip has excellent potential for wide applicability and verified that the strip methods for the optimized conditions are applicable to a variety of micropollutants. The lateral flow immunochromatographic strip could be used as a simple, rapid, and efficient screening tool for rapid on-site detection of a variety of micropollutants.

  2. Improved Analytical Sensitivity of Lateral Flow Assay using Sponge for HBV Nucleic Acid Detection.

    PubMed

    Tang, Ruihua; Yang, Hui; Gong, Yan; Liu, Zhi; Li, XiuJun; Wen, Ting; Qu, ZhiGuo; Zhang, Sufeng; Mei, Qibing; Xu, Feng

    2017-05-02

    Hepatitis B virus (HBV) infection is a serious public health problem, which can be transmitted through various routes (e.g., blood donation) and cause hepatitis, liver cirrhosis and liver cancer. Hence, it is necessary to do diagnostic screening for high-risk HBV patients in these transmission routes. Nowadays, protein-based technologies have been used for HBV testing, which however involve the issues of large sample volume, antibody instability and poor specificity. Nucleic acid hybridization-based lateral flow assay (LFA) holds great potential to address these limitations due to its low-cost, rapid, and simple features, but the poor analytical sensitivity of LFA restricts its application. In this study, we developed a low-cost, simple and easy-to-use method to improve analytical sensitivity by integrating sponge shunt into LFA to decrease the fluid flow rate. The thickness, length and hydrophobicity of the sponge shunt were sequentially optimized, and achieved 10-fold signal enhancement in nucleic acid testing of HBV as compared to the unmodified LFA. The enhancement was further confirmed by using HBV clinical samples, where we achieved the detection limit of 10 3 copies/ml as compared to 10 4 copies/ml in unmodified LFA. The improved LFA holds great potential for diseases diagnostics, food safety control and environment monitoring at point-of-care.

  3. Applying Chromatography.

    ERIC Educational Resources Information Center

    Klein, Jessie W.; Patev, Paul

    1998-01-01

    Presents three experiments to introduce students to different kinds of chromatography: (1) paper chromatography; (2) gel filtration chromatography; and (3) reverse-phase liquid chromatography. Written in the form of a laboratory manual, explanations of each of the techniques, materials needed, procedures, and a glossary are included. (PVD)

  4. Differentiating organically and conventionally grown oregano using ultraperformance liquid chromatography mass spectrometry (UPLC-MS), headspace gas chromatography with flame ionization detection (headspace-GC-FID), and flow injection mass spectrum (FIMS) fingerprints combined with multivariate data analysis.

    PubMed

    Gao, Boyan; Qin, Fang; Ding, Tingting; Chen, Yineng; Lu, Weiying; Yu, Liangli Lucy

    2014-08-13

    Ultraperformance liquid chromatography mass spectrometry (UPLC-MS), flow injection mass spectrometry (FIMS), and headspace gas chromatography (headspace-GC) combined with multivariate data analysis techniques were examined and compared in differentiating organically grown oregano from that grown conventionally. It is the first time that headspace-GC fingerprinting technology is reported in differentiating organically and conventionally grown spice samples. The results also indicated that UPLC-MS, FIMS, and headspace-GC-FID fingerprints with OPLS-DA were able to effectively distinguish oreganos under different growing conditions, whereas with PCA, only FIMS fingerprint could differentiate the organically and conventionally grown oregano samples. UPLC fingerprinting provided detailed information about the chemical composition of oregano with a longer analysis time, whereas FIMS finished a sample analysis within 1 min. On the other hand, headspace GC-FID fingerprinting required no sample pretreatment, suggesting its potential as a high-throughput method in distinguishing organically and conventionally grown oregano samples. In addition, chemical components in oregano were identified by their molecular weight using QTOF-MS and headspace-GC-MS.

  5. Lateral-flow colloidal gold-based immunoassay for the rapid detection of deoxynivalenol with two indicator ranges.

    PubMed

    Kolosova, Anna Yu; Sibanda, Liberty; Dumoulin, Frédéric; Lewis, Janet; Duveiller, Etienne; Van Peteghem, Carlos; De Saeger, Sarah

    2008-06-02

    A lateral-flow immunoassay using a colloidal gold-labelled monoclonal antibody was developed for the rapid detection of deoxynivalenol (DON). Different parameters, such as the amount of immunoreagents, type of the materials, composition of the blocking solution and of the detector reagent mixture, were investigated to provide the optimum assay performance. The experimental results demonstrated that such a visual test had an indicator range rather than a cut-off value. Thus, tests for DON determination with two different indicator ranges of 250-500 and 1000-2000 microg kg(-1) were designed. The method allowed detection of DON at low and high concentration levels, which could be useful for research and practical purposes. The assay applied to spiked wheat and pig feed samples demonstrated accurate and reproducible results. The applicability of the developed lateral-flow test was also confirmed under real field conditions. The test strips prepared in Belgium were sent to Mexico, where they were used for the screening of DON contamination in different bread wheat entries from Fusarium Head Blight inoculated plots. The results were compared with those obtained by ELISA and LC-MS/MS. A poor correlation between ELISA and LC-MS/MS was observed. Visual results of the dipstick tests were in a good agreement with the results of the LC-MS/MS method. Coupled with a simple and fast sample preparation, this qualitative one-step test based on the visual evaluation of results did not require any equipment. Results could be obtained within 10 min. The described assay format can be used as a simple, rapid, cost-effective and robust on-site screening tool for mycotoxin contamination in different agricultural commodities.

  6. Diagnosis of prosthetic joint infection with alpha-defensin using a lateral flow device: a multicentre study.

    PubMed

    Berger, P; Van Cauter, M; Driesen, R; Neyt, J; Cornu, O; Bellemans, J

    2017-09-01

    The purpose of this current multicentre study is to analyse the presence of alpha-defensin proteins in synovial fluid using the Synovasure lateral flow device and to determine its diagnostic reliability and accuracy compared with the prosthetic joint infection (PJI) criteria produced by the Musculoskeletal Infection Society (MSIS). A cohort of 121 patients comprising 85 total knee arthroplasties and 36 total hip arthroplasties was prospectively evaluated between May 2015 and June 2016 in three different orthopaedic centres. The tests were performed on patients with a chronically painful prosthesis undergoing a joint aspiration in a diagnostic pathway or during revision surgery. Based on the MSIS criteria, 34 patients (28%) would have had a PJI, and 87 patients had no PJI. Testing with the lateral flow device had a sensitivity of 97.1% (95% confidence intervals (CI) 84.5 to 99.9) and a specificity of 96.6% (95% CI 90.3 to 99.2). The positive predictive value was 91.7% (95% CI 77.7% to 98.3), and the negative predictive value was 98.8% (95% CI 93.6 to 99.9). Receiver operator characteristics analysis demonstrated an area under the curve for the Synovasure test of 0.97 (95% CI 0.93 to 1.00). Our findings suggest that the Synovasure test has an excellent diagnostic performance to confirm or reject the diagnosis of a PJI. The results are promising for the care of the painful or problematic knee and hip joint arthroplasty and the test should be considered as part of the diagnostic toolbox for PJIs. Cite this article: Bone Joint J 2017;99-B:1176-82. ©2017 The British Editorial Society of Bone & Joint Surgery.

  7. Intrinsic particle-induced lateral transport in microchannels

    PubMed Central

    Amini, Hamed; Sollier, Elodie; Weaver, Westbrook M.; Di Carlo, Dino

    2012-01-01

    In microfluidic systems at low Reynolds number, the flow field around a particle is assumed to maintain fore-aft symmetry, with fluid diverted by the presence of a particle, returning to its original streamline downstream. This current model considers particles as passive components of the system. However, we demonstrate that at finite Reynolds number, when inertia is taken into consideration, particles are not passive elements in the flow but significantly disturb and modify it. In response to the flow field, particles translate downstream while rotating. The combined effect of the flow of fluid around particles, particle rotation, channel confinement (i.e., particle dimensions approaching those of the channel), and finite fluid inertia creates a net recirculating flow perpendicular to the primary flow direction within straight channels that resembles the well-known Dean flow in curved channels. Significantly, the particle generating this flow remains laterally fixed as it translates downstream and only the fluid is laterally transferred. Therefore, as the particles remain inertially focused, operations can be performed around the particles in a way that is compatible with downstream assays such as flow cytometry. We apply this particle-induced transfer to perform fluid switching and mixing around rigid microparticles as well as deformable cells. This transport phenomenon, requiring only a simple channel geometry with no external forces to operate, offers a practical approach for fluid transfer at high flow rates with a wide range of applications, including sample preparation, flow reaction, and heat transfer. PMID:22761309

  8. Determination of alkylphenols by gas chromatography, elution liquid chromatography, and gel permeation chromatography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Wittmann, S.; Decsy, Z.; Regensperger, S.

    1984-01-01

    The separation and determination of groups of alkylphenols with C/sub 15/-C/sub 33/ isoalkane chains by gas chromatography, elution liquid chromatography, and gel permeation chromatography are described. Paraffinic hydrocarbons, monoalkylphenols, dialkylphenols, and bis(hydroxyphenyl)alkanes were identified in industrial alkylphenols by mass spectrometry. 7 references, 3 figures, 2 tables.

  9. Lateral fluid flow fractionation using dielectrophoresis (LFFF-DEP) for size-independent, label-free isolation of circulating tumor cells.

    PubMed

    Waheed, Waqas; Alazzam, Anas; Mathew, Bobby; Christoforou, Nicolas; Abu-Nada, Eiyad

    2018-06-15

    This short communication introduces a continuous-flow, dielectrophoresis-based lateral fluid flow fractionation microdevice for detection/isolation of circulating tumor cells in the presence of other haematological cells. The device utilizes two sets of planar interdigitated transducer electrodes micropatterned on top of a glass wafer using standard microfabrication techniques. A microchannel with a single inlet and two outlets, realized in polydimethylsiloxane, is bonded on the glass substrate. The two sets of electrodes slightly protrude into the microchannel. Both of the electrode sets are energized with signals at different frequencies and different operating voltages ensuring that the cancer cells experience positive dielectrophoretic force from one set of the electrodes and negative dielectrophoretic force from the other array. Normal cells experience unequal negative dielectrophoretic forces from opposing sets of electrodes. The resultant dielectrophoretic forces on cancer and normal cells push them to flow towards their designed outlets. Successful isolation of green fluorescent protein-labelled MDA-MB-231 breast cancer cells from regular blood cells, both suspended in a sucrose/dextrose medium, is reported in this work. Copyright © 2018 Elsevier B.V. All rights reserved.

  10. Point-of-care coagulation monitoring: first clinical experience using a paper-based lateral flow diagnostic device.

    PubMed

    Hegener, Michael A; Li, Hua; Han, Daewoo; Steckl, Andrew J; Pauletti, Giovanni M

    2017-09-01

    Vitamin K antagonists such as warfarin are the most widely used class of oral anticoagulants. Due to a narrow therapeutic window, patients on warfarin require regular monitoring. Self-testing using point-of-care (POC) diagnostic devices is available, but cost makes this monitoring method beyond reach for many. The main objective of this research was to assess the clinical utility of a low-cost, paper-based lateral flow POC diagnostic device developed for anticoagulation monitoring without the need for a separate electronic reader. Custom-fabricated lateral flow assay (LFA) test strips comprised of a glass fiber sample pad, a nitrocellulose analytical membrane, a cellulose wicking pad, and a plastic backing card were assembled in a plastic cassette. Healthy volunteers and patients on warfarin therapy were recruited for this prospective study. For each participant, a whole blood sample was collected via fingerstick to determine: (1) international normalized ratio (INR) using the CoaguChek® XS coagulometer, (2) hematocrit by centrifugation, and (3) red blood cell (RBC) travel distance on the experimental LFA device after 240 s using digital image analysis. RBC travel distance measured on the LFA device using blood samples obtained from warfarin patients positively correlated with increasing INR value and the LFA device had the capability to statistically distinguish between healthy volunteer INR values and those for patients groups with INR ≥ 2.6. From these data, it is predicted that this low-cost, paper-based LFA device can have clinical utility for identifying anticoagulated patients taking vitamin K antagonists who are outside of the desired therapeutic efficacy window.

  11. Inhibition of recombinase polymerase amplification by background DNA: a lateral flow-based method for enriching target DNA.

    PubMed

    Rohrman, Brittany; Richards-Kortum, Rebecca

    2015-02-03

    Recombinase polymerase amplification (RPA) may be used to detect a variety of pathogens, often after minimal sample preparation. However, previous work has shown that whole blood inhibits RPA. In this paper, we show that the concentrations of background DNA found in whole blood prevent the amplification of target DNA by RPA. First, using an HIV-1 RPA assay with known concentrations of nonspecific background DNA, we show that RPA tolerates more background DNA when higher HIV-1 target concentrations are present. Then, using three additional assays, we demonstrate that the maximum amount of background DNA that may be tolerated in RPA reactions depends on the DNA sequences used in the assay. We also show that changing the RPA reaction conditions, such as incubation time and primer concentration, has little effect on the ability of RPA to function when high concentrations of background DNA are present. Finally, we develop and characterize a lateral flow-based method for enriching the target DNA concentration relative to the background DNA concentration. This sample processing method enables RPA of 10(4) copies of HIV-1 DNA in a background of 0-14 μg of background DNA. Without lateral flow sample enrichment, the maximum amount of background DNA tolerated is 2 μg when 10(6) copies of HIV-1 DNA are present. This method requires no heating or other external equipment, may be integrated with upstream DNA extraction and purification processes, is compatible with the components of lysed blood, and has the potential to detect HIV-1 DNA in infant whole blood with high proviral loads.

  12. Does disseminated nontuberculous mycobacterial disease cause false-positive Determine B-LAM lateral flow assay results? A retrospective review.

    PubMed

    Nel, Jeremy S; Lippincott, Christopher K; Berhanu, Rebecca; Spencer, David C; Sanne, Ian M; Ive, Prudence

    2017-06-02

    We retrospectively reviewed the Determine TB-LAM lateral flow assay (LF-LAM) results among HIV-infected patients with disseminated nontuberculous mycobacterial (NTM) disease. LF-LAM was positive in 19/21 patients without evidence of tuberculosis coinfection. Although tuberculosis-NTM coinfection may have been underdiagnosed, our results suggest that disseminated NTM disease may cause false-positive LF-LAM results. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

  13. Influence of vertical and lateral heat transfer on permafrost thaw, peatland landscape transition, and groundwater flow

    USGS Publications Warehouse

    Kurylyk, Barret L.; Masaki, Masaki; Quinton, William L.; McKenzie, Jeffrey M.; Voss, Clifford I.

    2016-01-01

    Recent climate change has reduced the spatial extent and thickness of permafrost in many discontinuous permafrost regions. Rapid permafrost thaw is producing distinct landscape changes in the Taiga Plains of the Northwest Territories, Canada. As permafrost bodies underlying forested peat plateaus shrink, the landscape slowly transitions into unforested wetlands. The expansion of wetlands has enhanced the hydrologic connectivity of many watersheds via new surface and near-surface flow paths, and increased streamflow has been observed. Furthermore, the decrease in forested peat plateaus results in a net loss of boreal forest and associated ecosystems. This study investigates fundamental processes that contribute to permafrost thaw by comparing observed and simulated thaw development and landscape transition of a peat plateau-wetland complex in the Northwest Territories, Canada from 1970 to 2012. Measured climate data are first used to drive surface energy balance simulations for the wetland and peat plateau. Near-surface soil temperatures simulated in the surface energy balance model are then applied as the upper boundary condition to a three-dimensional model of subsurface water flow and coupled energy transport with freeze-thaw. Simulation results demonstrate that lateral heat transfer, which is not considered in many permafrost models, can influence permafrost thaw rates. Furthermore, the simulations indicate that landscape evolution arising from permafrost thaw acts as a positive feedback mechanism that increases the energy absorbed at the land surface and produces additional permafrost thaw. The modeling results also demonstrate that flow rates in local groundwater flow systems may be enhanced by the degradation of isolated permafrost bodies.

  14. LATERAL HEAT FLOW INFRARED THERMOGRAPHY FOR THICKNESS INDEPENDENT DETERMINATION OF THERMAL DIFFUSIVITY IN CFRP

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tralshawala, Nilesh; Howard, Don; Knight, Bryon

    2008-02-28

    In conventional infrared thermography, determination of thermal diffusivity requires thickness information. Recently GE has been experimenting with the use of lateral heat flow to determine thermal diffusivity without thickness information. This work builds on previous work at NASA Langley and Wayne State University but we incorporate thermal time of flight (tof) analysis rather than curve fitting to obtain quantitative information. We have developed appropriate theoretical models and a tof based data analysis framework to experimentally determine all components of thermal diffusivity from the time-temperature measurements. Initial validation was carried out using finite difference simulations. Experimental validation was done using anisotropicmore » carbon fiber reinforced polymer (CFRP) composites. We found that in the CFRP samples used, the in-plane component of diffusivity is about eight times larger than the through-thickness component.« less

  15. Detection of H5 and H7 highly pathogenic avian influenza virus with lateral flow devices: performance with healthy, sick and dead chickens

    USDA-ARS?s Scientific Manuscript database

    Rapid detection of highly pathogenic avian influenza virus (HPAIV) in the field is critical for effective disease control and to differentiate it from other diseases, such as Newcastle disease. Lateral flow devices (LFD) are commercially available and provide a fast, highly specific, on-site test fo...

  16. Development of a Smartphone-based reading system for lateral flow immunoassay.

    PubMed

    Lee, Sangdae; Kim, Giyoung; Moon, Jihea

    2014-11-01

    This study was conducted to develop and evaluate the performance of the Smartphone-based reading system for the lateral flow immunoassay (LFIA). Smartphone-based reading system consists of a Samsung Galaxy S2 Smartphone, Smartphone application, and a LFIA reader. LFIA reader is composed of the close-up lens with a focal length up to 30 mm, white LED light, lithium polymer battery, and main body. The Smartphone application for image acquisition and data analysis was developed on the Android platform. The standard curve was obtained by plotting the measured P(T)/P(c) or A(T)/A(c) ratio versus Salmonella standard concentration. The mean, standard deviation (SD), recovery, and relative standard deviation (RSD) were also calculated using additional experimental results. These data were compared with that obtained from the benchtop LFIA reader. The LOD in both systems was observed with 10(6) CFU/mL. The results show high accuracy and good reproducibility with a RSD less than 10% in the range of 10(6) to 10(9) CFU/mL. Due to the simple structure, good sensitivity, and high accuracy of the Smartphone-based reading system, this system can be substituted for the benchtop LFIA reader for point-of-care medical diagnostics.

  17. Comparison of antimicrobial peptide purification via free-flow electrophoresis and gel filtration chromatography.

    PubMed

    Xia, Zhi-Jun; Liu, Zhen; Kong, Fan-Zhi; Fan, Liu-Yin; Xiao, Hua; Cao, Cheng-Xi

    2017-12-01

    Antimicrobial peptides (AMPs) are usually small and cationic biomolecules with broad-spectrum antimicrobial activities against pathogens. Purifying them from complex samples is essential to study their physiochemical properties. In this work, free-flow zone electrophoresis (FFZE) was utilized to purify AMPs from yeast fermentation broth. Meanwhile, gel filtration chromatography (GFC) was conducted for comparison. The separation efficiency was evaluated by SDS-PAGE analysis of the fractions from both methods. Our results demonstrated as follows: (i) FFZE had more than 30-fold higher processing capacity as compared with GFC; (ii) FFZE could achieve 87% purity and 89% recovery rate while in GFC these parameters were about 93 and 82%, respectively; (iii) the former had ∼2-fold dilution but the latter had ∼13-fold dilution. Furthermore, Tricine-SDS-PAGE, Native-PAGE, and gel IEF were carried out to characterize the purified AMPs. We found that two peptides existed as a pair with the molecular mass of ∼5.5 and 7.0 kDa, while the same pI 7.8. These two peptides were proved to have the antimicrobial activity through the standardized agar diffusion method. Therefore, FFZE could be used to continuously purify AMPs with high bioactivity, which will lead to its wide application in the clinical and pharmaceutical fields. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Separation of Aeruginosin-865 from Cultivated Soil Cyanobacterium (Nostoc sp.) by Centrifugal Partition Chromatography combined with Gel Permeation Chromatography.

    PubMed

    Cheel, José; Minceva, Mirjana; Urajová, Petra; Aslam, Rabya; Hrouzek, Pavel; Kopecký, Jiří

    2015-10-01

    Aeruginosin-865 was isolated from cultivated soil cyanobacteria using a combination of centrifugal partition chromatography (CPC) and gel permeation chromatography. The solubility of Aer-865 in different solvents was evaluated using the conductor-like screening model for real solvents (COSMO-RS). The CPC separation was performed in descending mode with a biphasic solvent system composed of water-n-BuOH-acetic acid (5:4:1, v/v/v). The upper phase was used as a stationary phase, whereas the lower phase was employed as a mobile phase at a flow rate of 10 mL/min. The revolution speed and temperature of the separation column were 1700 rpm and 25 degrees C, respectively. Preparative CPC separation followed by gel permeation chromatography was performed on 50 mg of crude extract yielding Aer-865 (3.5 mg), with a purity over 95% as determined by HPLC. The chemical identity of the isolated compound was confirmed by comparing its spectroscopic data (UV, HRESI-MS, HRESI-MS/MS) with those of an authentic standard and data available in the literature.

  19. CdSe/ZnS Quantum Dot-Labeled Lateral Flow Strips for Rapid and Quantitative Detection of Gastric Cancer Carbohydrate Antigen 72-4

    NASA Astrophysics Data System (ADS)

    Yan, Xinyu; Wang, Kan; Lu, Wenting; Qin, Weijian; Cui, Daxiang; He, Jinghua

    2016-03-01

    Carbohydrate antigen 72-4 (CA72-4) is an important biomarker associated closely with diagnosis and prognosis of early gastric cancer. How to realize quick, sensitive, specific, and quantitative detection of CA72-4 in clinical specimens has become a great requirement. Herein, we reported a CdSe/ZnS quantum dot-labeled lateral flow test strip combined with a charge-coupled device (CCD)-based reader was developed for rapid, sensitive, and quantitative detection of CA72-4. Two mouse monoclonal antibodies (mAbs) against CA72-4 were employed. One of them was coated as a test line, while another mAb was labeled with quantum dots and coated onto conjugate pad. The goat anti-mouse IgG was immobilized as a control line. After sample was added, a sandwich structure was formed with CA72-4 and these two mAbs. The fluorescent signal from quantum dots (QD)-labeled mAb in sandwich structure was related to the amount of detected CA72-4. A CCD-based reader was used to realize quantitative detection of CA72-4. Results showed that developed QD-labeled lateral flow strips to detect CA72-4 biomarker with the sensitivity of 2 IU/mL and 10 min detection time. One hundred sera samples from clinical patients with gastric cancer and healthy people were used to confirm specificity of this strip method; results showed that established strip method own 100 % reproducibility and 100 % specificity compared with Roche electrochemiluminescence assay results. In conclusion, CdSe/ZnS quantum dot-labeled lateral flow strips for detection of CA72-4 could realize rapid, sensitive, and specific detection of clinical samples and could own great potential in clinical translation in near future.

  20. Significant viscosity dependent deviations from classical van Deemter theory in liquid chromatography with porous silica monolithic columns.

    PubMed

    Nesterenko, Pavel N; Rybalko, Marina A; Paull, Brett

    2005-06-01

    Significant deviations from classical van Deemter behaviour, indicative of turbulent flow liquid chromatography, has been recorded for mobile phases of varying viscosity on porous silica monolithic columns at elevated mobile phase flow rates.

  1. On-site detection of stacked genetically modified soybean based on event-specific TM-LAMP and a DNAzyme-lateral flow biosensor.

    PubMed

    Cheng, Nan; Shang, Ying; Xu, Yuancong; Zhang, Li; Luo, Yunbo; Huang, Kunlun; Xu, Wentao

    2017-05-15

    Stacked genetically modified organisms (GMO) are becoming popular for their enhanced production efficiency and improved functional properties, and on-site detection of stacked GMO is an urgent challenge to be solved. In this study, we developed a cascade system combining event-specific tag-labeled multiplex LAMP with a DNAzyme-lateral flow biosensor for reliable detection of stacked events (DP305423× GTS 40-3-2). Three primer sets, both event-specific and soybean species-specific, were newly designed for the tag-labeled multiplex LAMP system. A trident-like lateral flow biosensor displayed amplified products simultaneously without cross contamination, and DNAzyme enhancement improved the sensitivity effectively. After optimization, the limit of detection was approximately 0.1% (w/w) for stacked GM soybean, which is sensitive enough to detect genetically modified content up to a threshold value established by several countries for regulatory compliance. The entire detection process could be shortened to 120min without any large-scale instrumentation. This method may be useful for the in-field detection of DP305423× GTS 40-3-2 soybean on a single kernel basis and on-site screening tests of stacked GM soybean lines and individual parent GM soybean lines in highly processed foods. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Two-column sequential injection chromatography--new approach for fast and effective analysis and its comparison with gradient elution chromatography.

    PubMed

    Chocholous, Petr; Satínský, Dalibor; Sklenárová, Hana; Solich, Petr

    2010-05-23

    This work presents novel approach in low-pressure chromatography flow systems--two-column Sequential Injection Chromatography (2-C SIC) and its comparison with gradient elution chromatography on the same instrument. The system was equipped with two different chromatographic columns (connected to selection valve in parallel design) for isocratic separation and determination of all components in composed anti-inflammatory pharmaceutical preparation (tablets). The sample was first injected on the first column of length 30 mm where less retained analytes were separated and then the sample was injected on the second column of length 10 mm where more retained analytes were separated. The SIC system was based on a commercial SIChrom manifold (8-port high-pressure selection valve and medium-pressure syringe pump with 4 mL reservoir) (FIAlab, USA) with two commercially available monolithic columns the "first column" Chromolith Flash RP-18e (25 mm x 4.6 mm i.d. with guard column 5 mm x 4.6 mm i.d.) and the "second column" Chromolith RP-18e (10 mm x 4.6 mm i.d.) and CCD UV-vis detector USB 4000 with micro-volume 1.0 cm Z flow cell. Two mobile phases were used for analysis (one for each column). The mobile phase 1 used for elution of paracetamol, caffeine and salicylic acid (internal standard) was acetonitrile/water (10:90, v/v, the water part of pH 3.5 adjusted with acetic acid), flow rate was 0.9 mL min(-1) (volume 3.0 mL of mobile phase per analysis). The mobile phase 2 used for elution of propyphenazone was acetonitrile/water (30:70, v/v); flow rate was 1.2 mL min(-1) (volume 1.5 mL of mobile phase per analysis). Absorbance was monitored at 210 nm. Samples were prepared by dissolving of one tablet in 30% acetonitrile and 10 microL of filtered supernatant was injected on each column (2 x 10 microL). The chromatographic resolution between all compounds was >1.45 and analysis time was 5.5 min under the optimal conditions. Limits of detection were determined at 0.4 microg m

  3. Fully automated multifunctional ultrahigh pressure liquid chromatography system for advanced proteome analyses

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lee, Jung Hwa; Hyung, Seok-Won; Mun, Dong-Gi

    2012-08-03

    A multi-functional liquid chromatography system that performs 1-dimensional, 2-dimensional (strong cation exchange/reverse phase liquid chromatography, or SCX/RPLC) separations, and online phosphopeptides enrichment using a single binary nano-flow pump has been developed. With a simple operation of a function selection valve, which is equipped with a SCX column and a TiO2 (titanium dioxide) column, a fully automated selection of three different experiment modes was achieved. Because the current system uses essentially the same solvent flow paths, the same trap column, and the same separation column for reverse-phase separation of 1D, 2D, and online phosphopeptides enrichment experiments, the elution time information obtainedmore » from these experiments is in excellent agreement, which facilitates correlating peptide information from different experiments.« less

  4. Hierarchical CaCO3 chromatography: a stationary phase based on biominerals.

    PubMed

    Sato, Kosuke; Oaki, Yuya; Takahashi, Daisuke; Toshima, Kazunobu; Imai, Hiroaki

    2015-03-23

    In biomineralization, acidic macromolecules play important roles for the growth control of crystals through a specific interaction. Inspired by this interaction, we report on an application of the hierarchical structures in CaCO3 biominerals to a stationary phase of chromatography. The separation and purification of acidic small organic molecules are achieved by thin-layer chromatography and flash chromatography using the powder of biominerals as the stationary phase. The unit nanocrystals and their oriented assembly, the hierarchical structure, are suitable for the adsorption site of the target organic molecules and the flow path of the elution solvents, respectively. The separation mode is ascribed to the specific adsorption of the acidic molecules on the crystal face and the coordination of the functional groups to the calcium ions. The results imply that a new family of stationary phase of chromatography can be developed by the fine tuning of hierarchical structures in CaCO3 materials. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Giant oscillations of the current in a dirty 2D electron system flowing perpendicular to a lateral barrier under magnetic field

    NASA Astrophysics Data System (ADS)

    Kadigrobov, A. M.

    2017-08-01

    The charge transport in a dirty 2-dimensional electron system biased in the presence of a lateral potential barrier under magnetic field is theoretically studied. The quantum tunnelling across the barrier provides the quantum interference of the edge states localized on its both sides that results in giant oscillations of the charge current flowing perpendicular to the lateral junction. Our theoretical analysis is in a good agreement with the experimental observations presented in Kang et al. [Lett. Nat. 403, 59 (2000)]. In particular, positions of the conductance maxima coincide with the Landau levels while the conductance itself is essentially suppressed even at the energies at which the resonant tunnelling occurs and hence these puzzling observations can be resolved without taking into account the electron-electron interaction.

  6. High-pressure liquid chromatography with direct injection of gas sample.

    PubMed

    Astanin, Anton I; Baram, Grigory I

    2017-06-09

    The conventional method of using liquid chromatography to determine the composition of a gaseous mixture entails dissolving vapors in a suitable solvent, then obtaining a chromatograph of the resulting solution. We studied the direct introduction of a gaseous sample into a C18 reversed-phase column, followed by separation of the components by HPLC with UV detection. Since the chromatography was performed at high pressure, vapors readily dissolved in the eluent and the substances separated in the column as effectively as in liquid samples. Samples were injected into the column in two ways: a) through the valve without a flow stop; b) after stopping the flow and relieving all pressure. We showed that an injectable gas volume could reach 70% of column dead volume. When an injected gaseous sample volume was less than 10% of the column dead volume, the resulting peaks were symmetrical and the column efficiency was high. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Lateral jet injection into typical combustor flowfields

    NASA Technical Reports Server (NTRS)

    Lilley, D. G.

    1986-01-01

    The experimental problem of lateral jet injection into typical flow fields in the absence of combustion was studied. All flow fields being investigated have no expansion of the crossflow (the test section to swirler diameter ratio D/d = 1), after its passage through an optional swirler (with swirl vane angle phi = 0 (swirler removed), 45, and 70 degree). The lateral jet(s) is(are) located one test-section diameter downstream of the test-section inlet (x/D = 1). The lateral jets have round-sectioned nozzles, each of which has an area of 1/100th of the cross sectional area of the crossflow (A sub j/A sub c = 1/100). Jet-to-crossflow velocity ratios of R = v sub j/u sub o = 2, 4, and 6 were investigated. Helium-bubble low visualization, five-hole pitot probe time-mean velocity measurements, and single-wire time-mean velocity and normal and shear stress turbulence data were obtained in the research program.

  8. Preparative supercritical fluid chromatography: A powerful tool for chiral separations.

    PubMed

    Speybrouck, David; Lipka, Emmanuelle

    2016-10-07

    In 2012, the 4 biggest pharmaceutical blockbusters were pure enantiomers and separating racemic mixtures is now frequently a key step in the development of a new drug. For a long time, preparative liquid chromatography was the technique of choice for the separation of chiral compounds either during the drug discovery process to get up to a hundred grams of a pure enantiomer or during the clinical trial phases needing kilograms of material. However the advent of supercritical Fluid Chromatography (SFC) in the 1990s has changed things. Indeed, the use of carbon dioxide as the mobile phase in SFC offers many advantages including high flow rate, short equilibration time as well as low solvent consumption. Despite some initial teething troubles, SFC is becoming the primary method for preparative chiral chromatography. This article will cover recent developments in preparative SFC for the separation of enantiomers, reviewing several aspects such as instrumentation, chiral stationary phases, mobile phases or purely preparative considerations including overloading, productivity or large scale chromatography. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Supercritical fluid chromatography approach for a sustainable manufacture of new stereoisomeric anticancer agent.

    PubMed

    Ghinet, Alina; Zehani, Yasmine; Lipka, Emmanuelle

    2017-10-25

    Two routes aimed at the manufacture of unprecedented stereoisomeric combretastatin A-4 analogue were described: flash chromatography vs supercritical fluid chromatography. The latter has many advantages over liquid chromatography and was therefore chosen for the small scale separation of methyl 1-[(3-hydroxy-4-methoxyphenyl) (3,4,5-trimethoxyphenyl)methyl]-5-oxo-l-prolinate 5, with potential antitumoral activity. After a screening of six different polysaccharide based chiral stationary phases and four co-solvents, the percentage of co-solvent, the flow-rate and the outlet pressure were optimized through a design of experiments (DoE). The preparation of 50mg of each stereoisomer was achieved successfully on a Chiralpak AD-H with isopropanol as a co-solvent. Productivity (kkd), solvent usage and environmental factor (E Factor) were calculated. Flash chromatography and supercritical fluid chromatography approaches were compared in terms of yield and purity of each stereoisomer manufactured. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Liposome retention in size exclusion chromatography

    PubMed Central

    Ruysschaert, Tristan; Marque, Audrey; Duteyrat, Jean-Luc; Lesieur, Sylviane; Winterhalter, Mathias; Fournier, Didier

    2005-01-01

    Background Size exclusion chromatography is the method of choice for separating free from liposome-encapsulated molecules. However, if the column is not presaturated with lipids this type of chromatography causes a significant loss of lipid material. To date, the mechanism of lipid retention is poorly understood. It has been speculated that lipid binds to the column material or the entire liposome is entrapped inside the void. Results Here we show that intact liposomes and their contents are retained in the exclusion gel. Retention depends on the pore size, the smaller the pores, the higher the retention. Retained liposomes are not tightly fixed to the beads and are slowly released from the gels upon direct or inverted eluent flow, long washing steps or column repacking. Further addition of free liposomes leads to the elution of part of the gel-trapped liposomes, showing that the retention is transitory. Trapping reversibility should be related to a mechanism of partitioning of the liposomes between the stationary phase, water-swelled polymeric gel, and the mobile aqueous phase. Conclusion Retention of liposomes by size exclusion gels is a dynamic and reversible process, which should be accounted for to control lipid loss and sample contamination during chromatography. PMID:15885140

  11. A Better Method for Filling Pasteur Pipet Chromatography Columns

    ERIC Educational Resources Information Center

    Ruekberg, Ben

    2006-01-01

    An alternative method for the preparation of Pasteur pipet chromatography columns is presented that allows the column to be filled with solvent without bubbles and allows greater control of fluid flow while the materials to be separated are added. Students are required to wear gloves and goggles and caution should be used while handling glass…

  12. Sedentary behavior as a factor in determining lateral line contributions to rheotaxis.

    PubMed

    Bak-Coleman, Joseph; Coombs, Sheryl

    2014-07-01

    Rheotaxis is a robust, multisensory behavior with many potential benefits for fish and other aquatic animals. Visual (optic flow) cues appear to be sufficient for rheotaxis, but other sensory cues can clearly compensate for the loss of vision. The role of various non-visual sensory systems, in particular the flow-sensing lateral line, is poorly understood, largely because of widely varying methods and sensory conditions for studying rheotaxis. Here, we examine how sedentary behavior under visually deprived conditions affects the relative importance of lateral line cues in two species: one that is normally sedentary (the three-lined corydoras, Corydoras trilineatus) and one that normally swims continuously along the substrate (the blind cavefish, Astyanax mexicanus). No effect of lateral line disruption on rheotactic performance was found in blind cavefish, which were significantly more mobile than three-lined corydoras. By contrast, rheotaxis was significantly impaired at low, but not high, flow speeds in lateral-line-disabled corydoras. In addition, lateral-line-enabled corydoras were characterized by decreased mobility and increased rheotactic performance relative to lateral-line-disabled fish. Taken together, these results suggest that sedentary behavior is an important factor in promoting reliance on lateral line cues. © 2014. Published by The Company of Biologists Ltd.

  13. Hierarchical Nanogold Labels to Improve the Sensitivity of Lateral Flow Immunoassay

    NASA Astrophysics Data System (ADS)

    Serebrennikova, Kseniya; Samsonova, Jeanne; Osipov, Alexander

    2018-06-01

    Lateral flow immunoassay (LFIA) is a widely used express method and offers advantages such as a short analysis time, simplicity of testing and result evaluation. However, an LFIA based on gold nanospheres lacks the desired sensitivity, thereby limiting its wide applications. In this study, spherical nanogold labels along with new types of nanogold labels such as gold nanopopcorns and nanostars were prepared, characterized, and applied for LFIA of model protein antigen procalcitonin. It was found that the label with a structure close to spherical provided more uniform distribution of specific antibodies on its surface, indicative of its suitability for this type of analysis. LFIA using gold nanopopcorns as a label allowed procalcitonin detection over a linear range of 0.5-10 ng mL-1 with the limit of detection of 0.1 ng mL-1, which was fivefold higher than the sensitivity of the assay with gold nanospheres. Another approach to improve the sensitivity of the assay included the silver enhancement method, which was used to compare the amplification of LFIA for procalcitonin detection. The sensitivity of procalcitonin determination by this method was 10 times better the sensitivity of the conventional LFIA with gold nanosphere as a label. The proposed approach of LFIA based on gold nanopopcorns improved the detection sensitivity without additional steps and prevented the increased consumption of specific reagents (antibodies).

  14. Simultaneous Analysis of Monovalent Anions and Cations with a Sub-Microliter Dead-Volume Flow-Through Potentiometric Detector for Ion Chromatography

    PubMed Central

    Dumanli, Rukiye; Attar, Azade; Erci, Vildan; Isildak, Ibrahim

    2016-01-01

    A microliter dead-volume flow-through cell as a potentiometric detector is described in this article for sensitive, selective and simultaneous detection of common monovalent anions and cations in single column ion chromatography for the first time. The detection cell consisted of less selective anion- and cation-selective composite membrane electrodes together with a solid-state composite matrix reference electrode. The simultaneous separation and sensitive detection of sodium (Na+), potassium (K+), ammonium (NH4+), chloride (Cl−) and nitrate (NO3−) in a single run was achieved by using 98% 1.5 mM MgSO4 and 2% acetonitrile eluent with a mixed-bed ion-exchange separation column without suppressor column system. The separation and simultaneous detection of the anions and cations were completed in 6 min at the eluent flow-rate of 0.8 mL/min. Detection limits, at S/N = 3, were ranged from 0.2 to 1.0 µM for the anions and 0.3 to 3.0 µM for the cations, respectively. The developed method was successfully applied to the simultaneous determination of monovalent anions and cations in several environmental and biological samples. PMID:26786906

  15. Evaluation of a Cryptococcal antigen Lateral Flow Assay in serum and cerebrospinal fluid for rapid diagnosis of cryptococcosis in Colombia.

    PubMed

    Cáceres, Diego H; Zuluaga, Alejandra; Tabares, Ángela M; Chiller, Tom; González, Ángel; Gómez, Beatriz L

    2017-12-21

    A Lateral Flow Assay to detect cryptococcal antigen (CrAg® LFA) in serum and cerebrospinal fluid for the rapid diagnosis of cryptococcosis was evaluated. A retrospective validation was performed. Sensitivity and specificity of the CrAg® LFA was 100%. High concordance (kappa index=1.0) between Cryptococcal Antigen Latex Agglutination System (CALAS®) and CrAg® LFA was observed. CrAg® LFA showed higher analytical sensitivity for detecting low concentrations of cryptococcal antigen.

  16. Nonlinear Convective Flows in a Laterally Heated Two-Layer System with a Temperature-Dependent Heat Release/Consumption at the Interface

    NASA Astrophysics Data System (ADS)

    Simanovskii, Ilya; Viviani, Antonio; Dubois, Frank; Queeckers, Patrick

    2018-01-01

    Nonlinear convective flows developed under the joint action of buoyant and thermocapillary effects in a laterally heated two-layer system filling the closed cavity, have been investigated. The influence of a temperature-dependent interfacial heat release/consumption on nonlinear steady and oscillatory regimes, has been studied. It is shown that sufficiently strong temperature dependence of interfacial heat sinks and heat sources can change the sequence of bifurcations and lead to the development of specific oscillatory regimes in the system.

  17. Evaluation of a Cryptococcal antigen Lateral Flow Assay in serum and cerebrospinal fluid for rapid diagnosis of cryptococcosis in Colombia

    PubMed Central

    Cáceres, Diego H.; Zuluaga, Alejandra; Tabares, Ángela M.; Chiller, Tom; González, Ángel; Gómez, Beatriz L.

    2017-01-01

    ABSTRACT A Lateral Flow Assay to detect cryptococcal antigen (CrAg® LFA) in serum and cerebrospinal fluid for the rapid diagnosis of cryptococcosis was evaluated. A retrospective validation was performed. Sensitivity and specificity of the CrAg® LFA was 100%. High concordance (kappa index=1.0) between Cryptococcal Antigen Latex Agglutination System (CALAS®) and CrAg® LFA was observed. CrAg® LFA showed higher analytical sensitivity for detecting low concentrations of cryptococcal antigen. PMID:29267584

  18. Fine sediment trapping in river lateral cavities

    NASA Astrophysics Data System (ADS)

    Juez, C.; Maechler, G.; Schleiss, A. J.; Franca, M. J.

    2016-12-01

    River restoration is nowadays a major issue in the field of hydraulics. The natural course and geometry of the rivers have been artificially changed by human activities for different purposes (land gaining, flood protection, agriculture). From a morphologic point of view, channelized rivers often display a straight path and monotonous river banks. This is in contradiction with natural morphology, where a high diversity can be found across the channel path (meanders) and the banks (pools, riffles). One way to restore rivers consist of transforming the artificial banks by adding macro-roughness elements in the lateral river banks (also called cavities and lateral embayments). The creation of irregularities on the banks causes new flow patterns that diversify the river habitat. However, these lateral cavities may be also responsible of the change of the river morphology, since they may trap the fine sediments travelling within the water. This is particularly important in glacier-fed streams such as the upper Rhone River in Switzerland. These are charged with fine sediments resulting from the erosion of the underlying glaciers bottom. The creation of lateral cavities may affect the sediment and morphological equilibrium of the river since these may trap sediments. This work aims to study the influence of the lateral cavities on the transport of fine sediments in the main channel. A set of laboratory experiments were done which covered a wide range of rectangular cavity configurations. Key parameters such as the flow discharge, the aspect ratio of the cavities and the initial sediment concentration were tested. Surface PIV, sediment samples and turbidity temporal records were collected during the experiments. The trapping efficiency of the cavities and the associated flow patterns were analyzed. The resulting conclusions provide a useful information for the future design of river restoration projects.

  19. Instrumentation: Ion Chromatography.

    ERIC Educational Resources Information Center

    Fritz, James S.

    1987-01-01

    Discusses the importance of ion chromatography in separating and measuring anions. The principles of ion exchange are presented, along with some applications of ion chromatography in industry. Ion chromatography systems are described, as well as ion pair and ion exclusion chromatography, column packings, detectors, and programming. (TW)

  20. Automated on-line renewable solid-phase extraction-liquid chromatography exploiting multisyringe flow injection-bead injection lab-on-valve analysis.

    PubMed

    Quintana, José Benito; Miró, Manuel; Estela, José Manuel; Cerdà, Víctor

    2006-04-15

    In this paper, the third generation of flow injection analysis, also named the lab-on-valve (LOV) approach, is proposed for the first time as a front end to high-performance liquid chromatography (HPLC) for on-line solid-phase extraction (SPE) sample processing by exploiting the bead injection (BI) concept. The proposed microanalytical system based on discontinuous programmable flow features automated packing (and withdrawal after single use) of a small amount of sorbent (<5 mg) into the microconduits of the flow network and quantitative elution of sorbed species into a narrow band (150 microL of 95% MeOH). The hyphenation of multisyringe flow injection analysis (MSFIA) with BI-LOV prior to HPLC analysis is utilized for on-line postextraction treatment to ensure chemical compatibility between the eluate medium and the initial HPLC gradient conditions. This circumvents the band-broadening effect commonly observed in conventional on-line SPE-based sample processors due to the low eluting strength of the mobile phase. The potential of the novel MSFI-BI-LOV hyphenation for on-line handling of complex environmental and biological samples prior to reversed-phase chromatographic separations was assessed for the expeditious determination of five acidic pharmaceutical residues (viz., ketoprofen, naproxen, bezafibrate, diclofenac, and ibuprofen) and one metabolite (viz., salicylic acid) in surface water, urban wastewater, and urine. To this end, the copolymeric divinylbenzene-co-n-vinylpyrrolidone beads (Oasis HLB) were utilized as renewable sorptive entities in the micromachined unit. The automated analytical method features relative recovery percentages of >88%, limits of detection within the range 0.02-0.67 ng mL(-1), and coefficients of variation <11% for the column renewable mode and gives rise to a drastic reduction in operation costs ( approximately 25-fold) as compared to on-line column switching systems.

  1. Near-infrared fluorescence-based multiplex lateral flow immunoassay for the simultaneous detection of four antibiotic residue families in milk.

    PubMed

    Chen, Yiqiang; Chen, Qian; Han, Miaomiao; Liu, Jiangyang; Zhao, Peng; He, Lidong; Zhang, Yuan; Niu, Yiming; Yang, Wenjun; Zhang, Liying

    2016-05-15

    In this study, we developed a novel near-infrared fluorescence based multiplex lateral flow immunoassay by conjugating a near-infrared label to broad-specificity monoclonal antibody/receptor as detection complexes. Different antigens were dispensed onto separate test zones of nitrocellulose membrane to serve as capture reagents. This assay format allowed the simultaneous detection of four families of antibiotics (β-lactams, tetracyclines, quinolones and sulfonamides) in milk within 20 min. Qualitative and quantitative analysis of target antibiotics were realized by imaging the fluorescence intensity of the near-infrared label captured on respective test lines. For qualitative analysis, the cut-off values of β-lactams, tetracyclines, quinolones and sulfonamides were determined to be 8 ng/mL, 2 ng/mL, 4 ng/mL and 8 ng/mL respectively, which were much lower than the conventional gold nanoparticle based lateral flow immunoassay. For quantitative analysis, the detection ranges were 0.26-3.56 ng/mL for β-lactams, 0.04-0.98 ng/mL for tetracyclines, 0.08-2.0 ng/mL for quinolones, and 0.1-3.98 ng/mL for sulfonamides, with linear correlation coefficients higher than 0.97. The mean spiked recoveries ranged from 93.7% to 108.2% with coefficient of variations less than 16.3%. These results demonstrated that this novel immunoassay is a promising approach for rapidly screening the four families of antibiotic residues in milk. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Development of a recombinase polymerase amplification combined with lateral-flow dipstick assay for detection of bovine ephemeral fever virus.

    PubMed

    Hou, Peili; Zhao, Guimin; Wang, Hongmei; He, Chengqiang; Huan, Yanjun; He, Hongbin

    2018-04-01

    Bovine ephemeral fever virus (BEFV), identified as the causative pathogen of bovine ephemeral fever (BEF), is responsible for increasing numbers of epidemics/outbreaks and has a significant harmful effect on the livestock industry. Therefore, a rapid detection assay is imperative for BEFV diagnosis. In this study, we described the development of lateral-flow dipstick isothermal recombinase polymerase amplification (LFD-RPA) assays for detection of BEFV. RPA primers and LF probes were designed by targeting the specific G gene, and the amplification product can be visualized on a simple lateral flow dipstick with the naked eyes. The amplification reaction was performed at 38 °C for 20 min and LFD incubation time within 5 min. The detection limit of this assay was 8 copies per reaction, and there was no cross-reactivity with other bovine infectious viruses such as bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, bovine respiratory syncytial virus, bovine coronavirus, bovine parainfluenza virus type 3, bovine vesicular stomatitis virus. In addition, the assay was performed with total 128 clinical specimens and the diagnostic results were compared with conventional RT-PCR, real-time quantative(q) PCR. The result showed that the coincidence rate of BEFV LFD-RPA and real-time qPCR was 96.09% (123/128), which was higher than conventional RT-PCR. The RPA combined with LFD assay probably provides a rapid and sensitive alternative for diagnosis of BEFV infections outbreak. Copyright © 2018 Elsevier Ltd. All rights reserved.

  3. A Continuous Procedure Based on Column Chromatography to Purify Anthocyanins from Schisandra chinensis by a Macroporous Resin plus Gel Filtration Chromatography.

    PubMed

    Yue, Daran; Yang, Lei; Liu, Shouxin; Li, Jian; Li, Wei; Ma, Chunhui

    2016-02-06

    In our previous study, as natural food colorants and antioxidants, the color and content stabilities of Schisandra chinensis (S. chinensis) anthocyanins were investigated. In this work, the purification process parameters of S. chinensis anthocyanins using a macroporous resin and gel filtration chromatography were evaluated. The optimized parameters of static adsorption and desorption were as follows. The selected resin is HPD-300 (nonpolar copolymer styrene type resin), and the anthocyanins adsorption saturation capacity of HPD-300 resin was 0.475 mg/g dry resin. Adsorption time was 4 h, and 0.517 mg/mL of S. chinensis anthocyanins was adsorbed on the resin column with a flow rate of 39 mL/h (3 BV/h). After adsorption, the anthocyanins were completely desorpted with 2.5 BV of 90% (v/v) ethanol solution, and the desorption flow rate was 13 mL/h (1 BV/h). After purification by dynamic adsorption and desorption, the anthocyanins content in the effluent increased from 47.6 mg/g to 128.4 mg/g, the purity of anthocyanins increased six-fold from 5.08% to 30.43%, and the anthocyanins recovery was 96.5%. The major constituent of S. chinensis anthocyanins was isolated with Bio-Gel P2 gel filtration chromatography, and it was detected by liquid chromatography electrospray ionisation tandem mass spectrometry (LC-ESI-MS) as cyanidin-3-O-xylosylrutinoside. Moreover, the antioxidant activities of S. chinensis anthocyanins were investigated. After purification using the HPD-300 resin, the antioxidant activities of anthocyanins were increased 1.2-fold (FRAP) and 1.7-fold (ABTS).

  4. Rapid screening test for detection of oxytetracycline residues in milk using lateral flow assay.

    PubMed

    Naik, Laxmana; Sharma, Rajan; Mann, Bimlesh; Lata, Kiran; Rajput, Y S; Surendra Nath, B

    2017-03-15

    A rapid, semi-quantitative lateral flow assay (LFA) was developed to screen the oxytetracycline (OTC) antibiotics residues in milk samples. In this study a competitive immuno-assay format was established. Colloidal gold nano-particles (GNP) were prepared and used as labelling material in LFA. Polyclonal antibodies were generated against OTC molecule (anti-OTC), purified and the quality was assessed by enzyme linked immuno sorbet assay. For the first time membrane components required for LFA in milk system was optimized. GNP and anti-OTC stable conjugate preparation method was standardized, and then these components were placed over the conjugate pad. OTC coupled with carrier protein was placed on test line; species specific secondary antibodies were placed on the control line of the membrane matrix. Assay was validated by spiking OTC to antibiotic free milk samples and results could be accomplished within 5min. without need of any equipment. The visual detection limit was 30ppb. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Subsurface lateral preferential flow network revealed by time-lapse ground-penetrating radar in a hillslope

    NASA Astrophysics Data System (ADS)

    Guo, Li; Chen, Jin; Lin, Henry

    2014-12-01

    Subsurface lateral preferential flow (LPF) has been observed to contribute substantially to hillslope and catchment runoff. However, the complex nature of LPF and the lack of an appropriate investigation method have hindered direct LPF observation in the field. Thus, the initiation, persistence, and dynamics of LPF networks remain poorly understood. This study explored the application of time-lapse ground-penetrating radar (GPR) together with an artificial infiltration to shed light on the nature of LPF and its dynamics in a hillslope. Based on our enhanced field experimental setup and carefully refined GPR data postprocessing algorithms, we developed a new protocol to reconstruct LPF networks with centimeter resolution. This is the first time that a detailed LPF network and its dynamics have been revealed noninvasively along a hillslope. Real-time soil water monitoring and field soil investigation confirmed the locations of LPF mapped by time-lapse GPR surveys. Our results indicated the following: (1) Increased spatial variations of radar signals after infiltration suggested heterogeneous soil water changes within the studied soil, which reflected the generation and dynamics of LPF; (2) Two types of LPF networks were identified, the network at the location of soil permeability contrasts and that formed via a series of connected preferential flow paths; and (3) The formation and distribution of LPF networks were influenced by antecedent soil water condition. Overall, this study demonstrates clearly that carefully designed time-lapse GPR surveys with enhanced data postprocessing offer a practical and nondestructive way of mapping LPF networks in the field, thereby providing a potentially significant enhancement in our ability to study complex subsurface flow processes across the landscape.

  6. Model to Design Drip Hose Lateral Line

    NASA Astrophysics Data System (ADS)

    Ludwig, Rafael; Cury Saad, João Carlos

    2014-05-01

    Introduction The design criterion for non-pressure compensating drip hose is normally to have 10% of flow variation (Δq) in the lateral line, corresponding to 20% of head pressure variation (ΔH). Longer lateral lines in drip irrigation systems using conventional drippers provide cost reduction, but it is necessary to obtain to the uniformity of irrigation [1]. The use of Δq higher levels can provide longer lateral lines. [4] proposes the use of a 30% Δq and he found that this value resulted in distribution uniformity over 80%. [1] considered it is possible to extend the lateral line length using two emitters spacing in different section. He assumed that the spacing changing point would be at 40% of the total length, because this is approximately the location of the average flow according with [2]. [3] found that, for practical purposes, the average pressure is located at 40% of the length of the lateral line and that until this point it has already consumed 75% of total pressure head loss (hf ). In this case, the challenge for designers is getting longer lateral lines with high values of uniformity. Objective The objective of this study was to develop a model to design longer lateral lines using non-pressure compensating drip hose. Using the developed model, the hypotheses to be evaluated were: a) the use of two different spacing between emitters in the same lateral line allows longer length; b) it is possible to get longer lateral lines using high values of pressure variation in the lateral lines since the distribution uniformity stays below allowable limits. Methodology A computer program was developed in Delphi® based on the model developed and it is able to design lateral lines in level using non-pressure compensating drip hose. The input data are: desired distribution uniformity (DU); initial and final pressure in the lateral line; coefficients of relationship between emitter discharge and pressure head; hose internal diameter; pipe cross-sectional area

  7. Extensive database of liquid phase diffusion coefficients of some frequently used test molecules in reversed-phase liquid chromatography and hydrophilic interaction liquid chromatography.

    PubMed

    Song, Huiying; Vanderheyden, Yoachim; Adams, Erwin; Desmet, Gert; Cabooter, Deirdre

    2016-07-15

    Diffusion plays an important role in all aspects of band broadening in chromatography. An accurate knowledge of molecular diffusion coefficients in different mobile phases is therefore crucial in fundamental column performance studies. Correlations available in literature, such as the Wilke-Chang equation, can provide good approximations of molecular diffusion under reversed-phase conditions. However, these correlations have been demonstrated to be less accurate for mobile phases containing a large percentage of acetonitrile, as is the case in hydrophilic interaction liquid chromatography. A database of experimentally measured molecular diffusion coefficients of some 45 polar and apolar compounds that are frequently used as test molecules under hydrophilic interaction liquid chromatography and reversed-phase conditions is therefore presented. Special attention is given to diffusion coefficients of polar compounds obtained in large percentages of acetonitrile (>90%). The effect of the buffer concentration (5-10mM ammonium acetate) on the obtained diffusion coefficients is investigated and is demonstrated to mainly influence the molecular diffusion of charged molecules. Diffusion coefficients are measured using the Taylor-Aris method and hence deduced from the peak broadening of a solute when flowing through a long open tube. The validity of the set-up employed for the measurement of the diffusion coefficients is demonstrated by ruling out the occurrence of longitudinal diffusion, secondary flow interactions and extra-column effects, while it is also shown that radial equilibration in the 15m long capillary is effective. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Temperature gradient interaction chromatography of polymers: A molecular statistical model.

    PubMed

    Radke, Wolfgang; Lee, Sekyung; Chang, Taihyun

    2010-11-01

    A new model describing the retention in temperature gradient interaction chromatography of polymers is developed. The model predicts that polymers might elute in temperature gradient interaction chromatography in either an increasing or decreasing order or even nearly independent of molar mass, depending on the rate of the temperature increase relative to the flow rate. This is in contrast to solvent gradient elution, where polymers elute either in order of increasing molar mass or molar mass independent. The predictions of the newly developed model were verified with the literature data as well as new experimental data. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. A direct comparison of the performance of ground, beaded and silica-grafted MIPs in HPLC and turbulent flow chromatography applications.

    PubMed

    Fairhurst, Robert E; Chassaing, Christophe; Venn, Richard F; Mayes, Andrew G

    2004-12-15

    Spherical molecularly imprinted polymers (MIPs) specific to the beta-blocker propranolol have been synthesised using two different approaches and compared to traditional ground monolithic MIPs in HPLC and TFC applications. TFC is a LC technique used for rapid extraction of compounds directly from complex matrices. It can be easily coupled to HPLC and MS for automation of an extraction/analysis procedure. Spherical MIP beads were produced using a suspension polymerisation technique and silica/MIP composite beads by grafting MIP to spherical silica particles using a surface-bound initiator species. Synthesis of both beaded and silica-grafted MIPs was more practical than using the traditional grinding method and yields of spherical particles of the required size between 80 and 100% were routinely achieved. Under HPLC conditions, beaded and ground MIP materials showed a degree of chiral separation for all of the nine beta-blockers tested. The beaded MIP, however, showed much better flow properties and peak shape than the ground material. Silica-grafted MIP showed some separation in five of the drugs and a large improvement in peak shape and analysis times compared with both ground and beaded MIPs. The materials prepared were also used in extraction columns for Turbulent Flow Chromatography (TFC). Although no imprinting effect was observed under typical TFC conditions, beaded polymer materials showed promise for use as TFC extraction columns due to the good flow properties and clean extracts obtained.

  10. Characterizing pressure issues due to turbulent flow in tubing, in ultra-fast chiral supercritical fluid chromatography at up to 580bar.

    PubMed

    Berger, Terry A

    2016-12-02

    It has been widely suggested that the outlet pressure be changed to maintain constant density ("isopycnic" conditions) when comparing the kinetic performance of different columns in supercritical fluid chromatography (SFC). However, at high flow rates, flow in the tubing is turbulent, causing large extra-column pressure drops that limit options for changing outlet pressure. Some of these pressure drops occur before and some after the column, obscuring the actual column inlet and outlet pressures. In this work, a 4.6×100mm, 1.8μm R,R-Whelk-O1 column was used with low dispersion LD (120μm) plumbing to generate sub-1min chiral separations. However, the optimum, or near optimum, flow rate was 5mL-min -1 , producing a system pressure of 580bar (with 40% methanol, outlet pressure 120bar). Both the flow rate and pump pressure required were near the limits of the instrument, and significantly exceeded the capability of many other SFC's. Extra-column pressure drops (ΔP ec ) were as high as 200bar, caused mostly by turbulent flow in the tubing. The ΔP ec increased by more than the square of the flow rate. Reynolds Numbers (Re) were calculated for tubing as a function of flow rate between 100 and 400bar and 5-20% methanol in CO 2 , and 40°-60°C. This represents the most extensive analysis of turbulence in tubing in the SFC literature. Flow in 120μm ID tubing was calculated to be laminar below 1.0mL-min -1 , mostly transitional up to 2.5mL-min -1 and virtually always turbulent at 3mL-min -1 and higher. Flow in 170μm tubing is turbulent at lower flows but generates half the ΔP ec due to the lower mobile phase linear velocity. The results suggest that, while sub-minute chromatograms are easily generated, 4.6mm columns are not very user friendly for use with sub-2μm packings. The high flow rates required just to reach optimum result in high ΔP ec generated by the tubing, causing uncertainty in the true column inlet, outlet, and average column pressure/density. When

  11. Ultrasensitive lateral-flow assays based on quantum dot encapsulations with signal amplification

    NASA Astrophysics Data System (ADS)

    Li, Xue; Gong, Xiaoqun; Zhang, Bo; Liu, Yajuan; Chang, Jin; Zhang, Xuening

    2018-05-01

    Lateral-flow assays (LFAs), with its convenience and low cost, promise to become the in-home test format for early diagnosis and monitoring of tumor marker. However, the insufficient signal intensity was generated by signal reporters reducing the sensitivity of this format. In this study, a novel nanoscale signal reporter capable of amplifying the fluorescence signal is fabricated by encapsulating quantum dots (QDs) into modified tri-copolymer (poly(tert-butyl acrylate-co-ethyl acrylate-co-methacrylic acid)) (ODA- g-tri-copolymer). The amplified signal varied by simply adjusting the ratio of QDs to the ODA- g-tri-copolymer for obtaining QD nanospheres with high QD loading. They exhibits outstanding stability compared to the individual QDs both in the biological buffer and strong acid solutions. Here, human chorionic gonadotrophin (HCG) is employed as the model protein of LFAs. The results show that the detection limit of the QD nanospheres is pushed down to 0.016 IU/L, which is about 38.5 times enhanced compared to the individual QD-based LFAs without any signal amplifying. The ultrasensitive LFAs were attributed to the signal amplification strategy, and their efficiency and robustness demonstrated the great potential in clinical applications. [Figure not available: see fulltext.

  12. Review: Current applications and challenges for liquid chromatography coupled to isotope ratio mass spectrometry (LC/IRMS).

    PubMed

    Godin, Jean-Philippe; McCullagh, James S O

    2011-10-30

    High-precision isotope analysis is recognized as an essential research tool in many fields of study. Until recently, continuous flow isotope ratio mass spectrometry (CF-IRMS) was available via an elemental analyzer or a gas chromatography inlet system for compound-specific analysis of light stable isotopes. In 2004, however, an interface that coupled liquid chromatography with IRMS (LC/IRMS) became commercially available for the first time. This brought the capability for new areas of application, in particular enabling compound-specific δ(13)C analysis of non-volatile, aqueous soluble, compounds from complex mixtures. The interface design brought with it several analytical constraints, however, in particular a lack of compatibility with certain types of chromatography as well as limited flow rates and mobile phase compositions. Routine LC/IRMS methods have, however, been established for measuring the δ(13)C isotopic ratios of underivatized individual compounds for application in archeology, nutrition and physiology, geochemistry, hydrology, soil science and food authenticity. Seven years after its introduction, we review the technical advances and constraints, methodological developments and new applications of liquid chromatography coupled to isotope ratio mass spectrometry. Copyright © 2011 John Wiley & Sons, Ltd.

  13. Identification and quantification of odours from oxobiodegradable polyethylene oxidised under a free radical flow by headspace solid-phase microextraction followed by gas chromatography-olfactometry-mass spectrometry.

    PubMed

    Wrona, Magdalena; Vera, Paula; Pezo, Davinson; Nerín, Cristina

    2017-09-01

    Recently oxobiodegradable polyethylene gained popularity as food packaging material due to its potential to reduce polymer waste. However, this type of material can release after its oxidation off-odour compounds that affect the organoleptic properties of packaged food. Odour compounds released from both polyethylene and oxobiodegradable polyethylene before and after oxidation under a free radicals flow were investigated after 1 day, 2 days and 3 days of oxidation. The samples were analysed using headspace solid phase microextraction followed by gas chromatography-mass spectrometry and headspace solid phase microextraction coupled to gas chromatography-olfactometry-mass spectrometry. Sixty-two different odorous compounds were identified. 4-methylthio-2-butanone (fruit), nonanal (fat) and 3,6-nonadienal (fat) were present in different materials before oxidation. Multiple headspace-solid phase microextraction has been used to quantify all analytes. The most abundant compound was (Z)-3-hexenyl hexanoate with a concentration range between 1.5791±0.1387µg/g and 4.8181±0.3123µg/g. Compounds such as 2-dodecenal, 2-octenal, 2-pentanol, 3-nonenal, 3,6-nonadienal, ethyl 3-methylbutanoate, ethyl octenoate, hexanone, isopropyl hexanoate, octanal were below their LOD evaluated using MS detector; however, they were detected by gas chromatography-olfactometry. The minimum LOD and LOQ were 0.011µg/g and 0.036µg/g, respectively. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Chromatography resin support

    DOEpatents

    Dobos, James G.

    2002-01-01

    An apparatus and method of using an improved chromatography resin support is disclosed. The chromatography support platform is provided by a stainless steel hollow cylinder adapted for being inserted into a chromatography column. An exterior wall of the stainless steel cylinder defines a groove for carrying therein an "O"-ring. The upper surface of the stainless steel column is covered by a fine stainless steel mesh welded to the edges of the stainless steel cylinder. When placed upon a receiving ledge defined within a chromatography column, the "O"-ring provides a fluid tight seal with the inner edge wall of the chromatography cylinder. The stainless steel mesh supports the chromatography matrix and provides a back flushable support which is economical and simple to construct.

  15. Thin-Layer Chromatography Experiments That Illustrate General Problems in Chromatography.

    ERIC Educational Resources Information Center

    Lederer, M.; Leipzig-Pagani, E.

    1996-01-01

    Describes experiments that illustrate a number of general principles such as pattern identification, displacement chromatography, and salting-out adsorption, plus an experiment that demonstrates that identification by chromatography alone is impossible. Illustrates that chromatography is still possible with quite simple means, notwithstanding the…

  16. Instrumentation for analytical scale supercritical fluid chromatography.

    PubMed

    Berger, Terry A

    2015-11-20

    Analytical scale supercritical fluid chromatography (SFC) is largely a sub-discipline of high performance liquid chromatography (HPLC), in that most of the hardware and software can be used for either technique. The aspects that separate the 2 techniques stem from the use of carbon dioxide (CO2) as the main component of the mobile phase in SFC. The high compressibility and low viscosity of CO2 mean that pumps, and autosamplers designed for HPLC either need to be modified or an alternate means of dealing with compressibility needs to be found. The inclusion of a back pressure regulator and a high pressure flow cell for any UV-Vis detector are also necessary. Details of the various approaches, problems and solutions are described. Characteristics, such as adiabatic vs. isothermal compressibility, thermal gradients, and refractive index issues are dealt with in detail. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Development of a Rapid Immunochromatographic Lateral Flow Device Capable of Differentiating Phytase Expressed from Recombinant Aspergillus niger phyA2 and Genetically Modified Corn.

    PubMed

    Zhou, Xiaojin; Hui, Elizabeth; Yu, Xiao-Lin; Lin, Zhen; Pu, Ling-Kui; Tu, Zhiguan; Zhang, Jun; Liu, Qi; Zheng, Jian; Zhang, Juan

    2015-05-06

    Phytase is a phosphohydrolase considered highly specific for the degradation of phytate to release bound phosphorus for animal consumption and aid in the reduction of environmental nutrient loading. New sources of phytase have been sought that are economically and efficiently productive including the construction of genetically modified (GM) phytase products designed to bypass the costs associated with feed processing. Four monoclonal antibodies (EH10a, FA7, AF9a, and CC1) raised against recombinant Aspergillus niger phyA2 were used to develop a highly specific and sensitive immunochromatographic lateral flow device for rapid detection of transgenic phytase, such as in GM corn. Antibodies sequentially paired and tested along lateral flow strips showed that the EH10a-FA7 antibody pair was able to detect the recombinant yeast-phytase at 5 ng/mL, whereas the AF9a-CC1 antibody pair to GM phytase corn was able to detect at 2 ng/mL. Concurrent to this development, evidence was revealed which suggests that antibody binding sites may be glycosylated.

  18. Automation of flow injection gas diffusion-ion chromatography for the nanomolar determination of methylamines and ammonia in seawater and atmospheric samples

    PubMed Central

    Gibb, Stuart W.; Wood, John W.; Fauzi, R.; Mantoura, C.

    1995-01-01

    The automation and improved design and performance of Flow Injection Gas Diffusion-Ion Chromatography (FIGD-IC), a novel technique for the simultaneous analysis of trace ammonia (NH3) and methylamines (MAs) in aqueous media, is presented. Automated Flow Injection Gas Diffusion (FIGD) promotes the selective transmembrane diffusion of MAs and NH3 from aqueous sample under strongly alkaline (pH > 12, NaOH), chelated (EDTA) conditions into a recycled acidic acceptor stream. The acceptor is then injected onto an ion chromatograph where NH3 and the MAs are fully resolved as their cations and detected conductimetrically. A versatile PC interfaced control unit and data capture unit (DCU) are employed in series to direct the selonoid valve switching sequence, IC operation and collection of data. Automation, together with other modifications improved both linearily (R2 > 0.99 MAs 0-100 nM, NH3 0-1000 nM) and precision (<8%) of FIGD-IC at nanomolar concentrations, compared with the manual procedure. The system was successfully applied to the determination of MAs and NH3 in seawater and in trapped particulate and gaseous atmospheric samples during an oceanographic research cruise. PMID:18925047

  19. The Role of Nanoparticle Design in Determining Analytical Performance of Lateral Flow Immunoassays.

    PubMed

    Zhan, Li; Guo, Shuang-Zhuang; Song, Fayi; Gong, Yan; Xu, Feng; Boulware, David R; McAlpine, Michael C; Chan, Warren C W; Bischof, John C

    2017-12-13

    Rapid, simple, and cost-effective diagnostics are needed to improve healthcare at the point of care (POC). However, the most widely used POC diagnostic, the lateral flow immunoassay (LFA), is ∼1000-times less sensitive and has a smaller analytical range than laboratory tests, requiring a confirmatory test to establish truly negative results. Here, a rational and systematic strategy is used to design the LFA contrast label (i.e., gold nanoparticles) to improve the analytical sensitivity, analytical detection range, and antigen quantification of LFAs. Specifically, we discovered that the size (30, 60, or 100 nm) of the gold nanoparticles is a main contributor to the LFA analytical performance through both the degree of receptor interaction and the ultimate visual or thermal contrast signals. Using the optimal LFA design, we demonstrated the ability to improve the analytical sensitivity by 256-fold and expand the analytical detection range from 3 log 10 to 6 log 10 for diagnosing patients with inflammatory conditions by measuring C-reactive protein. This work demonstrates that, with appropriate design of the contrast label, a simple and commonly used diagnostic technology can compete with more expensive state-of-the-art laboratory tests.

  20. Determination of perfluorinated chemicals in food and drinking water using high-flow solid-phase extraction and ultra-high performance liquid chromatography/tandem mass spectrometry.

    PubMed

    Chang, Ying-Chia; Chen, Wen-Ling; Bai, Fang-Yu; Chen, Pau-Chung; Wang, Gen-Shuh; Chen, Chia-Yang

    2012-01-01

    For this study, we developed methods of determining ten perfluorinated chemicals in drinking water, milk, fish, beef, and pig liver using high-flow automated solid-phase extraction (SPE) and ultra-high performance liquid chromatography/tandem mass spectrometry. The analytes were separated on a core-shell Kinetex C18 column. The mobile phase was composed of methanol and 10-mM N-methylmorpholine. Milk was digested with 0.5 N potassium hydroxide in Milli-Q water, and was extracted with an Atlantic HLB disk to perform automated SPE at a flow rate ranged from 70 to 86 mL/min. Drinking water was directly extracted by the SPE. Solid food samples were digested in alkaline methanol and their supernatants were diluted and also processed by SPE. The disks were washed with 40% methanol/60% water and then eluted with 0.1% ammonium hydroxide in methanol. Suppression of signal intensity of most analytes by matrixes was lower than 50%; it was generally lower in fish and drinking water but higher in liver. Most quantitative biases and relative standard deviations were lower than 15%. The limits of detection for most analytes were sub-nanograms per liter for drinking water and sub-nanograms per gram for solid food samples. This method greatly shortened the time and labor needed for digestion, SPE, and liquid chromatography. This method has been applied to analyze 14 types of food samples. Perfluorooctanoic acid was found to be the highest among the analytes (median at 3.2-64 ng/g wet weight), followed by perfluorodecanoic acid (0.7-25 ng/g) and perfluorododecanoic acid (0.6-15 ng/g).

  1. Pressurized planar electrochromatography, high-performance thin-layer chromatography and high-performance liquid chromatography--comparison of performance.

    PubMed

    Płocharz, Paweł; Klimek-Turek, Anna; Dzido, Tadeusz H

    2010-07-16

    Kinetic performance, measured by plate height, of High-Performance Thin-Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC) and Pressurized Planar Electrochromatography (PPEC) was compared for the systems with adsorbent of the HPTLC RP18W plate from Merck as the stationary phase and the mobile phase composed of acetonitrile and buffer solution. The HPLC column was packed with the adsorbent, which was scrapped from the chromatographic plate mentioned. An additional HPLC column was also packed with adsorbent of 5 microm particle diameter, C18 type silica based (LiChrosorb RP-18 from Merck). The dependence of plate height of both HPLC and PPEC separating systems on flow velocity of the mobile phase and on migration distance of the mobile phase in TLC system was presented applying test solute (prednisolone succinate). The highest performance, amongst systems investigated, was obtained for the PPEC system. The separation efficiency of the systems investigated in the paper was additionally confirmed by the separation of test component mixture composed of six hormones. 2010 Elsevier B.V. All rights reserved.

  2. Evaluation of a novel helium ionization detector within the context of (low-)flow modulation comprehensive two-dimensional gas chromatography.

    PubMed

    Franchina, Flavio A; Maimone, Mariarosa; Sciarrone, Danilo; Purcaro, Giorgia; Tranchida, Peter Q; Mondello, Luigi

    2015-07-10

    The present research is focused on the use and evaluation of a novel helium ionization detector, defined as barrier discharge ionization detector (BID), within the context of (low-)flow modulation comprehensive two-dimensional gas chromatography (FM GC×GC). The performance of the BID device was compared to that of a flame ionization detector (FID), under similar FM GC×GC conditions. Following development and optimization of the FM GC×GC method, the BID was subjected to fine tuning in relation to acquisition frequency and discharge flow. Moreover, the BID performance was measured and compared to that of the FID, in terms of extra-column band broadening, sensitivity and dynamic range. The comparative study was carried out by using standard compounds belonging to different chemical classes, along with a sample of diesel fuel. Advantages and disadvantages of the BID system, also within the context of FM GC×GC, are critically discussed. In general, the BID system was characterized by a more limited dynamic range and increased sensitivity, compared to the FID. Additionally, BID and FID contribution to band broadening was found to be similar under the operational conditions applied. Particular attention was devoted to the behaviour of the FM GC×GC-BID system toward saturated and aromatic hydrocarbons, for a possible future use in the field of mineral-oil food contamination research. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Development of Lateral Flow Immunochromatographic Strips for Micropollutants Screening Using Colorants of Aptamer Functionalized Nanogold Particles Part I: Methodology and Optimization.

    PubMed

    Zhao, Shuai; Zhang, Shan; Wang, Sai; Liu, Jiahui; Dong, Yiyang

    2018-05-03

    A methodology of lateral flow immunochromatographic strip based on aptamer was developed for on-site detectionof the small molecule micropollutants. In the present study, we try for the first time to investigate the feasibility of developing a strip assay for the analysis of micropollutants as methodological prototypes by combining the high selectivity and affinity of aptamers with the unique optical properties of nanogolds. This quantitative method was based on the competition for the aptamer between targets and DNA probes. Crucial parameters that might influence the sensitivity, such as the size of nanogolds, amount of aptamer, type and pH of streptavidin, type of nitrocellulose (NC) membrane, blocking procedure, and reading time, were systematically investigated to obtain the optimum assay performance. With the optimized conditions [nanogolds 25 nm, 50 μM aptamer, pH 8 of GSA (a type of streptavidin named "SA Gold," which is a sulfhydrylization streptavidin), Millipore HFC 135 NC membrane, 1% bovine serum albumin as the blocking agent and added in the running buffer and sample pad soakage agents, and 20 min reading time] the aptamer-based lateral flow assay will show a low visual limit of detection and scanning reader LOD. The strip for on-site screening using colorants of aptamer functionalized nanogold particles did not require any complicated equipment and was a potential portable tool for rapid identification of micropollutants.

  4. Nepem-211 ion exchange conductive membrane immobilized tris(2,2´-bipyridyl) ruthenium(II) electrogenerated chemiluminescence flow sensor for high-performance liquid chromatography and its application.

    PubMed

    Li, Yongbo; Zhang, Zhujun

    2013-01-01

    We developed a sensitive and robust electrogenerated chemiluminescence (ECL) flow sensor based on Ru(bpy)3(2+) immobilized with a Nepem-211 perfluorinated ion exchange conductance membrane, which has robustness and stability under a wide range of chemical and physical conditions, good electrical conductivity, isotropy and a high exchange capacity for immobilization of Ru(bpy)3(2+). The flow sensor has been used as a post-column detector in high-performance liquid chromatography for determination of erythromycin and clarithromycin in honey and pork, and tricyclic antidepressant drugs in human urine. Under optimal conditions, the linear ranges were 0.03-26 ng/μL and 0.01-1 ng/μL for macrolides and tricyclic antidepressant drugs, respectively. The detection limits were 0.02, 0.01, 0.01, 0.06 and 0.003 ng/μL for erythromycin, clarithromycin, doxepin, amitriptyline and clomipramine, respectively. There is no post-column reagent addition. In addition to the conservation expensive reagents, the experimental setup was simplified. The flow sensor was used for 2 years with high sensitivity and stability. Copyright © 2013 John Wiley & Sons, Ltd.

  5. Lateral Downflows in Sunspot Penumbral Filaments and their Temporal Evolution

    NASA Astrophysics Data System (ADS)

    Esteban Pozuelo, S.; Bellot Rubio, L. R.; de la Cruz Rodríguez, J.

    2015-04-01

    We study the temporal evolution of downflows observed at the lateral edges of penumbral filaments in a sunspot located very close to the disk center. Our analysis is based on a sequence of nearly diffraction-limited scans of the Fe i 617.3 nm line taken with the CRisp Imaging Spectro-Polarimeter instrument at the Swedish 1 m Solar Telescope. We compute Dopplergrams from the observed intensity profiles using line bisectors and filter the resulting velocity maps for subsonic oscillations. Lateral downflows appear everywhere in the center-side penumbra as small, weak patches of redshifts next to or along the edges of blueshifted flow channels. These patches have an intermittent life and undergo mergings and fragmentations quite frequently. The lateral downflows move together with the hosting filaments and react to their shape variations, very much resembling the evolution of granular convection in the quiet Sun. There is a good relation between brightness and velocity in the center-side penumbra, with downflows being darker than upflows on average, which is again reminiscent of quiet Sun convection. These results point to the existence of overturning convection in sunspot penumbrae, with elongated cells forming filaments where the flow is upward but very inclined, and weak lateral downward flows. In general, the circular polarization profiles emerging from the lateral downflows do not show sign reversals, although sometimes we detect three-lobed profiles that are suggestive of opposite magnetic polarities in the pixel.

  6. Biochemical separations by continuous-bed chromatography.

    PubMed

    Tisch, T L; Frost, R; Liao, J L; Lam, W K; Remy, A; Scheinpflug, E; Siebert, C; Song, H; Stapleton, A

    1998-08-07

    Innovations in column-packing media for biomolecule purification have progressed from large spherical, porous polysaccharide beads to advanced polymeric supports. Continuous-bed technology is a radical new technology for chromatography based on the polymerization of advanced monomers and ionomers directly in the chromatographic column. The polymer chains form aggregates which coalesce into a dense, homogeneous network of interconnected nodules consisting of microparticles with an average diameter of 3000 A. The voids or channels between the nodules are large enough to permit a high hydrodynamic flow. Due to the high cross-linking of the polymer matrix, the surface of each nodule is nonporous yet the polymeric microparticles provide a very large surface area for high binding capacity. This paper will demonstrate the properties and advantages of using a continuous bed support for high resolution biomolecule separations at high flow-rates without sacrificing capacity.

  7. Hydroxylapatite chromatography.

    PubMed

    Broadhurst, A V

    2001-05-01

    Hydroxylapatite (also called hydroxyapatite), a form of calcium phosphate, can be used as a matrix for the chromatography of both proteins and nucleic acids. Protocols are provided for both standard low-pressure chromatography of a protein mixture using a hydroxylapatite column prepared in the laboratory, and an HPLC method, applicable to proteins and nucleic acids, that uses a commercially available column. Alternate protocols describe column chromatography using a step gradient or batch binding and step-gradient elution.

  8. Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for detection of Toxoplasma gondii in the environment.

    PubMed

    Wu, Y D; Xu, M J; Wang, Q Q; Zhou, C X; Wang, M; Zhu, X Q; Zhou, D H

    2017-08-30

    Toxoplasma gondii infects all warm-blooded vertebrates, resulting in a great threat to human health and significant economic loss to the livestock industry. Ingestion of infectious oocysts of T. gondii from the environment is the major source of transmission. Detection of T. gondii oocysts by existing methods is laborious, time-consuming and expensive. The objective of the present study was to develop a recombinase polymerase amplification (RPA) method combined with a lateral flow (LF) strip for detection of T. gondii oocysts in the soil and water. The DNA of T. gondii oocysts was amplified by a pair of specific primers based on the T. gondii B1 gene over 15min at a constant temperature ranging from 30°C to 45°C using RPA. The amplification product was visualized by the lateral flow (LF) strip within 5min using the specific probe added to the RPA reaction system. The sensitivity of the established assay was 10 times higher than that of nested PCR with a lower detection limit of 0.1 oocyst per reaction, and there was no cross-reactivity with other closely related protozoan species. Fifty environmental samples were further assessed for the detection validity of the LF-RPA assay (B1-LF-RPA) and compared with nested PCR based on the B1 gene sequence. The B1-LF-RPA and nested PCR both showed that 5 out of the 50 environmental samples were positive. The B1-LF-RPA method was also proven to be sufficiently tolerant of existing inhibitors in the environment. In addition, the advantages of simple operation, speediness and cost-effectiveness make B1-LF-RPA a promising molecular detection tool for T. gondii. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Blood coagulation screening using a paper-based microfluidic lateral flow device.

    PubMed

    Li, H; Han, D; Pauletti, G M; Steckl, A J

    2014-10-21

    A simple approach to the evaluation of blood coagulation using a microfluidic paper-based lateral flow assay (LFA) device for point-of-care (POC) and self-monitoring screening is reported. The device utilizes whole blood, without the need for prior separation of plasma from red blood cells (RBC). Experiments were performed using animal (rabbit) blood treated with trisodium citrate to prevent coagulation. CaCl2 solutions of varying concentrations are added to citrated blood, producing Ca(2+) ions to re-establish the coagulation cascade and mimic different blood coagulation abilities in vitro. Blood samples are dispensed into a paper-based LFA device consisting of sample pad, analytical membrane and wicking pad. The porous nature of the cellulose membrane separates the aqueous plasma component from the large blood cells. Since the viscosity of blood changes with its coagulation ability, the distance RBCs travel in the membrane in a given time can be related to the blood clotting time. The distance of the RBC front is found to decrease linearly with increasing CaCl2 concentration, with a travel rate decreasing from 3.25 mm min(-1) for no added CaCl2 to 2.2 mm min(-1) for 500 mM solution. Compared to conventional plasma clotting analyzers, the LFA device is much simpler and it provides a significantly larger linear range of measurement. Using the red colour of RBCs as a visible marker, this approach can be utilized to produce a simple and clear indicator of whether the blood condition is within the appropriate range for the patient's condition.

  10. Time distribution of adsorption entropy of gases on heterogeneous surfaces by reversed-flow gas chromatography.

    PubMed

    Katsanos, Nicholas A; Kapolos, John; Gavril, Dimitrios; Bakaoukas, Nicholas; Loukopoulos, Vassilios; Koliadima, Athanasia; Karaiskakis, George

    2006-09-15

    The reversed-flow gas chromatography (RF-GC) technique has been applied to measure the adsorption entropy over time, when gaseous pentane is adsorbed on the surface of two solids (gamma-alumina and a silica supported rhodium catalyst) at 393.15 and 413.15K, respectively. Utilizing experimental chromatographic data, this novel methodology also permits the simultaneous measurement of the local adsorption energy, epsilon, local equilibrium adsorbed concentration, c(s)(*), and local adsorption isotherm, theta(p, T, epsilon) in a time resolved way. In contrast with other inverse gas chromatographic methods, which determine the standard entropy at zero surface coverage, the present method operates over a wide range of surface coverage taking into account not only the adsorbate-adsorbent interaction, but also the adsorbate-adsorbate interaction. One of the most interesting observations of the present work is the fact that the interaction of n-pentane is spontaneous on the Rh/SiO(2) catalyst for a very short time interval compared to that on gamma-Al(2)O(3). This can explain the different kinetic behavior of each particular gas-solid system, and it can be attributed to the fact that large amounts of n-C(5)H(12) are present on the active sites of the Rh/SiO(2) catalyst compared to those on gamma-Al(2)O(3), as the local equilibrium adsorbed concentration values, c(s)(*), indicate.

  11. Direction of unsaturated flow in a homogeneous and isotropic hillslope

    USGS Publications Warehouse

    Lu, Ning; Kaya, Basak Sener; Godt, Jonathan W.

    2011-01-01

    The distribution of soil moisture in a homogeneous and isotropic hillslope is a transient, variably saturated physical process controlled by rainfall characteristics, hillslope geometry, and the hydrological properties of the hillslope materials. The major driving mechanisms for moisture movement are gravity and gradients in matric potential. The latter is solely controlled by gradients of moisture content. In a homogeneous and isotropic saturated hillslope, absent a gradient in moisture content and under the driving force of gravity with a constant pressure boundary at the slope surface, flow is always in the lateral downslope direction, under either transient or steady state conditions. However, under variably saturated conditions, both gravity and moisture content gradients drive fluid motion, leading to complex flow patterns. In general, the flow field near the ground surface is variably saturated and transient, and the direction of flow could be laterally downslope, laterally upslope, or vertically downward. Previous work has suggested that prevailing rainfall conditions are sufficient to completely control these flow regimes. This work, however, shows that under time-varying rainfall conditions, vertical, downslope, and upslope lateral flow can concurrently occur at different depths and locations within the hillslope. More importantly, we show that the state of wetting or drying in a hillslope defines the temporal and spatial regimes of flow and when and where laterally downslope and/or laterally upslope flow occurs.

  12. Direction of unsaturated flow in a homogeneous and isotropic hillslope

    USGS Publications Warehouse

    Lu, N.; Kaya, B.S.; Godt, J.W.

    2011-01-01

    The distribution of soil moisture in a homogeneous and isotropic hillslope is a transient, variably saturated physical process controlled by rainfall characteristics, hillslope geometry, and the hydrological properties of the hillslope materials. The major driving mechanisms for moisture movement are gravity and gradients in matric potential. The latter is solely controlled by gradients of moisture content. In a homogeneous and isotropic saturated hillslope, absent a gradient in moisture content and under the driving force of gravity with a constant pressure boundary at the slope surface, flow is always in the lateral downslope direction, under either transient or steady state conditions. However, under variably saturated conditions, both gravity and moisture content gradients drive fluid motion, leading to complex flow patterns. In general, the flow field near the ground surface is variably saturated and transient, and the direction of flow could be laterally downslope, laterally upslope, or vertically downward. Previous work has suggested that prevailing rainfall conditions are sufficient to completely control these flow regimes. This work, however, shows that under time-varying rainfall conditions, vertical, downslope, and upslope lateral flow can concurrently occur at different depths and locations within the hillslope. More importantly, we show that the state of wetting or drying in a hillslope defines the temporal and spatial regimes of flow and when and where laterally downslope and/or laterally upslope flow occurs. Copyright 2011 by the American Geophysical Union.

  13. Flowing atmospheric pressure afterglow combined with laser ablation for direct analysis of compounds separated by thin-layer chromatography.

    PubMed

    Cegłowski, Michał; Smoluch, Marek; Reszke, Edward; Silberring, Jerzy; Schroeder, Grzegorz

    2016-01-01

    A thin-layer chromatography-mass spectrometry (TLC-MS) setup for characterization of low molecular weight compounds separated on standard TLC plates has been constructed. This new approach successfully combines TLC separation, laser ablation, and ionization using flowing atmospheric pressure afterglow (FAPA) source. For the laser ablation, a low-priced 445-nm continuous-wave diode laser pointer, with a power of 1 W, was used. The combination of the simple, low-budget laser pointer and the FAPA ion source has made this experimental arrangement broadly available, also for small laboratories. The approach was successfully applied for the characterization of low molecular weight compounds separated on TLC plates, such as a mixture of pyrazole derivatives, alkaloids (nicotine and sparteine), and an extract from a drug tablet consisting of paracetamol, propyphenazone, and caffeine. The laser pointer used was capable of ablating organic compounds without the need of application of any additional substances (matrices, staining, etc.) on the TLC spots. The detection limit of the proposed method was estimated to be 35 ng/cm(2) of a pyrazole derivative.

  14. Lateral blasts at Mount St. Helens and hazard zonation

    USGS Publications Warehouse

    Crandell, D.R.; Hoblitt, R.P.

    1986-01-01

    Lateral blasts at andesitic and dacitic volcanoes can produce a variety of direct hazards, including ballistic projectiles which can be thrown to distances of at least 10 km and pyroclastic density flows which can travel at high speed to distances of more than 30 km. Indirect effect that may accompany such explosions include wind-borne ash, pyroclastic flows formed by the remobilization of rock debris thrown onto sloping ground, and lahars. Two lateral blasts occurred at a lava dome on the north flank of Mount St. Helens about 1200 years ago; the more energetic of these threw rock debris northeastward across a sector of about 30?? to a distance of at least 10 km. The ballistic debris fell onto an area estimated to be 50 km2, and wind-transported ash and lapilli derived from the lateral-blast cloud fell on an additional lobate area of at least 200 km2. In contrast, the vastly larger lateral blast of May 18, 1980, created a devastating pyroclastic density flow that covered a sector of as much as 180??, reached a maximum distance of 28 km, and within a few minutes directly affected an area of about 550 km2. The May 18 lateral blast resulted from the sudden, landslide-induced depressurization of a dacite cryptodome and the hydrothermal system that surrounded it within the volcano. We propose that lateral-blast hazard assessments for lava domes include an adjoining hazard zone with a radius of at least 10 km. Although a lateral blast can occur on any side of a dome, the sector directly affected by any one blast probably will be less than 180??. Nevertheless, a circular hazard zone centered on the dome is suggested because of the difficulty of predicting the direction of a lateral blast. For the purpose of long-term land-use planning, a hazard assessment for lateral blasts caused by explosions of magma bodies or pressurized hydrothermal systems within a symmetrical volcano could designate a circular potential hazard area with a radius of 35 km centered on the volcano

  15. High-pressure size exclusion chromatography analysis of dissolved organic matter isolated by tangential-flow ultra filtration

    USGS Publications Warehouse

    Everett, C.R.; Chin, Y.-P.; Aiken, G.R.

    1999-01-01

    A 1,000-Dalton tangential-flow ultrafiltration (TFUF) membrane was used to isolate dissolved organic matter (DOM) from several freshwater environments. The TFUF unit used in this study was able to completely retain a polystyrene sulfonate 1,800-Dalton standard. Unaltered and TFUF-fractionated DOM molecular weights were assayed by high-pressure size exclusion chromatography (HPSEC). The weight-averaged molecular weights of the retentates were larger than those of the raw water samples, whereas the filtrates were all significantly smaller and approximately the same size or smaller than the manufacturer-specified pore size of the membrane. Moreover, at 280 nm the molar absorptivity of the DOM retained by the ultrafilter is significantly larger than the material in the filtrate. This observation suggests that most of the chromophoric components are associated with the higher molecular weight fraction of the DOM pool. Multivalent metals in the aqueous matrix also affected the molecular weights of the DOM molecules. Typically, proton-exchanged DOM retentates were smaller than untreated samples. This TFUF system appears to be an effective means of isolating aquatic DOM by size, but the ultimate size of the retentates may be affected by the presence of metals and by configurational properties unique to the DOM phase.

  16. Fully automated screening of veterinary drugs in milk by turbulent flow chromatography and tandem mass spectrometry

    PubMed Central

    Stolker, Alida A. M.; Peters, Ruud J. B.; Zuiderent, Richard; DiBussolo, Joseph M.

    2010-01-01

    There is an increasing interest in screening methods for quick and sensitive analysis of various classes of veterinary drugs with limited sample pre-treatment. Turbulent flow chromatography in combination with tandem mass spectrometry has been applied for the first time as an efficient screening method in routine analysis of milk samples. Eight veterinary drugs, belonging to seven different classes were selected for this study. After developing and optimising the method, parameters such as linearity, repeatability, matrix effects and carry-over were studied. The screening method was then tested in the routine analysis of 12 raw milk samples. Even without internal standards, the linearity of the method was found to be good in the concentration range of 50 to 500 µg/L. Regarding repeatability, RSDs below 12% were obtained for all analytes, with only a few exceptions. The limits of detection were between 0.1 and 5.2 µg/L, far below the maximum residue levels for milk set by the EU regulations. While matrix effects—ion suppression or enhancement—are obtained for all the analytes the method has proved to be useful for screening purposes because of its sensitivity, linearity and repeatability. Furthermore, when performing the routine analysis of the raw milk samples, no false positive or negative results were obtained. PMID:20379812

  17. Liquid Crystals in Chromatography

    NASA Astrophysics Data System (ADS)

    Witkiewicz, Zygfryd

    The following sections are included: * INTRODUCTION * LIQUID CRYSTALS SUITABLE FOR GAS CHROMATOGRAPHY * Monomeric Liquid Crystal Stationary Phases * Polymeric Liquid Crystal Stationary Phases * Polymeric Liquid Crystal Stationary Phases * Conventional Analytical Columns * Capillary Columns * FACTORS AFFECTING THE CHROMATOGRAPHIC SEPARATIONS ON LIQUID CRYSTAL STATIONARY PHASES * Kind of Mesophase of the Liquid Crystal * Molecular Structure of the Liquid Crystals and of the Chromatographed Substances * Substrate on which the Liquid Crystal is Deposited * ANALYTICAL APPLICATIONS OF LIQUID CRYSTAL STATIONARY PHASES IN GAS CHROMATOGRAPHY * Separation of Isomers of Benzene and Naphthalene Derivatives * Separation of Alkane and Alkene Isomers * Separation of Mixtures of Benzene and Aliphatic Hydrocarbon Derivatives Containing Heteroatoms * Separation of Polynuclear Hydrocarbons * INVESTIGATION OF THE PROPERTIES OF LIQUID CRYSTALS BY GAS CHROMATOGRAPHY * APPLICATION OF LIQUID CRYSTALS IN LIQUID CHROMATOGRAPHY * Column Chromatography * Thin-Layer Chromatography * APPLICATION OF LIQUID CRYSTAL STATIONARY PHASES IN SUPERCRITICAL FLUID CHROMATOGRAPHY * FINAL REMARKS * References

  18. Coupling flash liquid chromatography with mass spectrometry for enrichment and isolation of milk oligosaccharides for functional studies.

    PubMed

    Strum, John S; Aldredge, Danielle; Barile, Daniela; Lebrilla, Carlito B

    2012-05-15

    Mass spectrometry has been coupled with flash liquid chromatography to yield new capabilities for isolating nonchromophoric material from complicated biological mixtures. A flash liquid chromatography/tandem mass spectrometry (LC/MS/MS) method enabled fraction collection of milk oligosaccharides from biological mixtures based on composition and structure. The method is compatible with traditional gas pressure-driven flow flash chromatography widely employed in organic chemistry laboratories. The online mass detector enabled real-time optimization of chromatographic parameters to favor separation of oligosaccharides that would otherwise be indistinguishable from coeluting components with a nonspecific detector. Unlike previously described preparative LC/MS techniques, we have employed a dynamic flow connection that permits any flow rate from the flash system to be delivered from 1 to 200 ml/min without affecting the ionization conditions of the mass spectrometer. A new way of packing large amounts of graphitized carbon allowed the enrichment and separation of milligram quantities of structurally heterogeneous mixtures of human milk oligosaccharides (HMOs) and bovine milk oligosaccharides (BMOs). Abundant saccharide components in milk, such as lactose and lacto-N-tetraose, were separated from the rarer and less abundant oligosaccharides that have greater structural diversity and biological functionality. Neutral and acidic HMOs and BMOs were largely separated and enriched with a dual binary solvent system. Published by Elsevier Inc.

  19. Comparison of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography for the separation of synthetic cathinones.

    PubMed

    Carnes, Stephanie; O'Brien, Stacey; Szewczak, Angelica; Tremeau-Cayel, Lauriane; Rowe, Walter F; McCord, Bruce; Lurie, Ira S

    2017-09-01

    A comparison of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography for the separation of synthetic cathinones has been conducted. Nine different mixtures of bath salts were analyzed in this study. The three different chromatographic techniques were examined using a general set of controlled synthetic cathinones as well as a variety of other synthetic cathinones that exist as positional isomers. Overall 35 different synthetic cathinones were analyzed. A variety of column types and chromatographic modes were examined for developing each separation. For the ultra high performance supercritical fluid chromatography separations, analyses were performed using a series of Torus and Trefoil columns with either ammonium formate or ammonium hydroxide as additives, and methanol, ethanol or isopropanol organic solvents as modifiers. Ultra high performance liquid chromatographic separations were performed in both reversed phase and hydrophilic interaction chromatographic modes using SPP C18 and SPP HILIC columns. Gas chromatography separations were performed using an Elite-5MS capillary column. The orthogonality of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography was examined using principal component analysis. For the best overall separation of synthetic cathinones, the use of ultra high performance supercritical fluid chromatography in combination with gas chromatography is recommended. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Separation techniques: Chromatography

    PubMed Central

    Coskun, Ozlem

    2016-01-01

    Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. Proteins can be purified based on characteristics such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the stationary phase. Four separation techniques based on molecular characteristics and interaction type use mechanisms of ion exchange, surface adsorption, partition, and size exclusion. Other chromatography techniques are based on the stationary bed, including column, thin layer, and paper chromatography. Column chromatography is one of the most common methods of protein purification. PMID:28058406

  1. Development and validation of a turbulent flow chromatography and tandem mass spectrometry method for the quantitation of methotrexate and its metabolites 7-hydroxy methotrexate and DAMPA in serum

    PubMed Central

    Schofield, Ryan C.; Ramanathan, Lakshmi V.; Murata, Kazunori; Grace, Marie; Fleisher, Martin; Pessin, Melissa S.; Carlow, Dean C.

    2016-01-01

    A rapid and simple turbulent flow liquid chromatography (TFC–LC) method implementing positive heated electrospray ionization (HESI) for the accurate and precise determination of methotrexate (MTX), 7-hydroxy methotrexate (7-OH MTX), and 4-amino-4-deoxy-N10-methylpteroic acid (DAMPA) concentrations in serum was developed. MTX was isolated from serum samples (100 μL) after protein precipitation with methanol containing formic acid and internal standard (MTX-D3) followed by centrifugation. The supernatant was injected into the turbulent flow liquid chromatography which is followed by electrospray positive ionization tandem mass spectrometry (TFC–LC–MS/MS) and quantified using a six-point calibration curve. For MTX and DAMPA the assays were linear from 10 to 1000 nmol/L and for 7-OH MTX from 20 to 2000 nmol/L. Dilutions of 10, 100 and 1000-fold were validated giving a clinically reportable range of 10 nmol/L to 5 × 105 nmol/L. Within-day and between-day precisions at concentrations spanning the analytical measurement ranges were less than 10% for all three analytes. MTX, DAMPA and 7-OH MTX were sufficiently stable under all relevant analytical conditions. No significant matrix effect was observed during the method validation. The TFC–LC-MS/MS MTX method was also compared with three other clinically validated MTX assays: a dihydrofolate reductase (DHFR) inhibition assay, an immunoassay based on fluorescence polarization and a previously developed LC–MS/MS assay. PMID:26322588

  2. Gas Chromatography.

    ERIC Educational Resources Information Center

    Karasek, Francis W.; And Others

    1984-01-01

    This review covers fundamental developments in gas chromatography during 1982 and 1983. Literature is considered under these headings: columns; liguid phases; solid supports; sorption processes and solvents; open tubular column gas chromatography; instrumentation; high-resolution columns and applications; other techniques; qualitative and…

  3. Rapid Detection of Listeria by Bacteriophage Amplification and SERS-Lateral Flow Immunochromatography

    PubMed Central

    Stambach, Nicholas R.; Carr, Stephanie A.; Cox, Christopher R.; Voorhees, Kent J.

    2015-01-01

    A rapid Listeria detection method was developed utilizing A511 bacteriophage amplification combined with surface-enhanced Raman spectroscopy (SERS) and lateral flow immunochromatography (LFI). Anti-A511 antibodies were covalently linked to SERS nanoparticles and printed onto nitrocellulose membranes. Antibody-conjugated SERS nanoparticles were used as quantifiable reporters. In the presence of A511, phage-SERS nanoparticle complexes were arrested and concentrated as a visible test line, which was interrogated quantitatively by Raman spectroscopy. An increase in SERS intensity correlated to an increase in captured phage-reporter complexes. SERS limit of detection was 6 × 106 pfu·mL−1, offering detection below that obtainable by the naked eye (LOD 6 × 107 pfu·mL−1). Phage amplification experiments were carried out at a multiplicity of infection (MOI) of 0.1 with 4 different starting phage concentrations monitored over time using SERS-LFI and validated by spot titer assay. Detection of L. monocytogenes concentrations of 1 × 107 colony forming units (cfu)·mL−1, 5 × 106 cfu·mL−1, 5 × 105 cfu·mL−1 and 5 × 104 cfu·mL−1 was achieved in 2, 2, 6, and 8 h, respectively. Similar experiments were conducted at a constant starting phage concentration (5 × 105 pfu·mL−1) with MOIs of 1, 2.5, and 5 and were detected in 2, 4, and 5 h, respectively. PMID:26694448

  4. Development of quantitative radioactive methodologies on paper to determine important lateral-flow immunoassay parameters.

    PubMed

    Mosley, Garrett L; Nguyen, Phuong; Wu, Benjamin M; Kamei, Daniel T

    2016-08-07

    The lateral-flow immunoassay (LFA) is a well-established diagnostic technology that has recently seen significant advancements due in part to the rapidly expanding fields of paper diagnostics and paper-fluidics. As LFA-based diagnostics become more complex, it becomes increasingly important to quantitatively determine important parameters during the design and evaluation process. However, current experimental methods for determining these parameters have certain limitations when applied to LFA systems. In this work, we describe our novel methods of combining paper and radioactive measurements to determine nanoprobe molarity, the number of antibodies per nanoprobe, and the forward and reverse rate constants for nanoprobe binding to immobilized target on the LFA test line. Using a model LFA system that detects for the presence of the protein transferrin (Tf), we demonstrate the application of our methods, which involve quantitative experimentation and mathematical modeling. We also compare the results of our rate constant experiments with traditional experiments to demonstrate how our methods more appropriately capture the influence of the LFA environment on the binding interaction. Our novel experimental approaches can therefore more efficiently guide the research process for LFA design, leading to more rapid advancement of the field of paper-based diagnostics.

  5. A Numerical Model of Exchange Chromatography Through 3D Lattice Structures

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Salloum, Maher; Robinson, David B.

    Rapid progress in the development of additive manufacturing technologies is opening new opportunities to fabricate structures that control mass transport in three dimensions across a broad range of length scales. We describe a structure that can be fabricated by newly available commercial 3D printers. It contains an array of regular three-dimensional flow paths that are in intimate contact with a solid phase, and thoroughly shuffle material among the paths. We implement a chemically reacting flow model to study its behavior as an exchange chromatography column, and compare it to an array of one-dimensional flow paths that resemble more traditional honeycombmore » monoliths. A reaction front moves through the columns and then elutes. Here, the front is sharper at all flow rates for the structure with three-dimensional flow paths, and this structure is more robust to channel width defects than the one-dimensional array.« less

  6. A Numerical Model of Exchange Chromatography Through 3D Lattice Structures

    DOE PAGES

    Salloum, Maher; Robinson, David B.

    2018-01-30

    Rapid progress in the development of additive manufacturing technologies is opening new opportunities to fabricate structures that control mass transport in three dimensions across a broad range of length scales. We describe a structure that can be fabricated by newly available commercial 3D printers. It contains an array of regular three-dimensional flow paths that are in intimate contact with a solid phase, and thoroughly shuffle material among the paths. We implement a chemically reacting flow model to study its behavior as an exchange chromatography column, and compare it to an array of one-dimensional flow paths that resemble more traditional honeycombmore » monoliths. A reaction front moves through the columns and then elutes. Here, the front is sharper at all flow rates for the structure with three-dimensional flow paths, and this structure is more robust to channel width defects than the one-dimensional array.« less

  7. A multiplex protein-free lateral flow assay for detection of microRNAs based on unmodified molecular beacons.

    PubMed

    Javani, Atefeh; Javadi-Zarnaghi, Fatemeh; Rasaee, Mohammad Javad

    2017-11-15

    Lateral flow assays (LFAs) have promising potentials for point-of-care applications. Recently, many LFAs have been reported that are based on hybridization of oligonucleotide strands. Mostly, biotinylated capture DNAs are immobilized on the surface of a nitrocellulose membrane via streptavidin interactions. During the assay, stable colorful complexes get formed that are visible by naked eyes. Here, we present an inexpensive and unique design of LFA that applies unmodified oligonucleotides at capture lines. The presented LFA do not utilize streptavidin or any other affinity protein. We employ structural switch of molecular beacons (MB) in combination with base stacking hybridization (BSH) phenomenon. The unique design of the reported LFA provided high selectivity for target oligonucleotides. We validated potential applications of the system for detection of DNA mimics of two microRNAs in multiplex assays. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Mobile Phone Sensing of Cocaine in a Lateral Flow Assay Combined with a Biomimetic Material.

    PubMed

    Guler, Emine; Yilmaz Sengel, Tulay; Gumus, Z Pinar; Arslan, Mustafa; Coskunol, Hakan; Timur, Suna; Yagci, Yusuf

    2017-09-19

    Lateral flow assays (LFAs) are an ideal choice for drug abuse testing favored by their practicability, portability, and rapidity. LFA based on-site rapid screening devices provide positive/negative judgment in a short response time. The conventionally applied competitive assay format used for small molecule analysis such as abused drugs restricts the quantitation ability of LFA strips. We report herein, for the first time, a new strategy using the noncompetitive assay format via a biomimetic material, namely, poly(p-phenylene) β-cyclodextrin poly(ethylene glycol) (PPP-CD-g-PEG) combined with gold nanoparticle (AuNP) conjugates as the labeling agent to recognize the target cocaine molecule in the test zone. The intensities of the visualized red color in the test line indicate that the cocaine concentrations were analyzed via a smartphone application. Significantly, a combination of this platform with a smartphone application provides quantitative data on the cocaine amount, making it a very inventive and attractive approach especially for on-site applications at critical points such as traffic stops and the workplace.

  9. Rapid Identification of OXA-48 and OXA-163 Subfamilies in Carbapenem-Resistant Gram-Negative Bacilli with a Novel Immunochromatographic Lateral Flow Assay.

    PubMed

    Pasteran, Fernando; Denorme, Laurence; Ote, Isabelle; Gomez, Sonia; De Belder, Denise; Glupczynski, Youri; Bogaerts, Pierre; Ghiglione, Barbara; Power, Pablo; Mertens, Pascal; Corso, Alejandra

    2016-11-01

    We assessed a novel immunochromatographic lateral flow assay for direct identification of OXA-48-like carbapenemases and accurate differentiation of allele variants with distinct substrate profiles (OXA-48 or OXA-163 subfamilies). The assay allowed rapid (less than 4 min) and reliable direct confirmation of OXA-163- and/or OXA-48-like enzymes (with 100% sensitivity and 100% specificity) from cultured colonies that were recovered from both solid medium and spiked blood culture bottles. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  10. Ecosystem processes at the watershed scale: hydrologic vegetation gradient as an indicator for lateral hydrologic connectivity of headwater catchments

    Treesearch

    Taehee Hwang; James M. Vose; Christina Tague

    2012-01-01

    Lateral water flow in catchments can produce important patterns in water and nutrient fluxes and stores and also influences the long-term spatial development of forest ecosystems. Specifically, patterns of vegetation type and density along hydrologic flow paths can represent a signal of the redistribution of water and nitrogen mediated by lateral hydrologic flow. This...

  11. Modified RS-LAMP assay and use of lateral flow devices for rapid detection of Leifsonia xyli subsp. xyli.

    PubMed

    Naidoo, N; Ghai, M; Moodley, K; Mkize, L; Martin, L; McFarlane, S; Rutherford, S

    2017-12-01

    Ratoon stunt (RS) caused by bacterium Leifsonia xyli subsp. xyli (Lxx) results in substantial yield losses in sugarcane (Saccharum sp. L. hybrid). Since RS does not produce reliable symptoms in the field, laboratory-based techniques are necessary for detection. Loop-mediated isothermal amplification (LAMP) assay overcomes the limitations of laboratory-based techniques which are costly, time consuming and cannot be used for near-field detection. A sensitive LAMP assay was developed to detect Lxx at 65°C in 30 min. However, carry-over contamination affected the reliability of the assay. In the present study, contaminants were successfully eliminated by incorporation of uracil nucleoside glycosylase (1 U μl -1 ) into the LAMP assay and incubation for 10 min at 37°C. To avoid the use of colorimetric reagents, lateral flow devices were successfully used for the detection of LAMP products and were equally sensitive to detection by agarose gel electrophoresis. The use of exudate from leaf sheath discs as an alternate template for the LAMP assay was found to be less sensitive than xylem sap. The preprepared master mix could be stored for up to 4 months at -20°C without any reduction in performance. These changes make the assay suitable for near-field detection in laboratories with basic facilities. This study presents a modified loop-mediated isothermal amplification (LAMP) assay for the detection of Leifsonia xyli subsp. xyli. Modifications include incorporation of uracil nucleoside glycosylase to eliminate carry-over contamination and substitution of colorimetric detection for the use of lateral flow devices. LAMP master mix was preprepared and was stably stored up to 4 months at -20°C. Sugarcane leaf sheaths worked well as a substitute to xylem sap as template, although the sensitivity was lower. These modifications allow the assay to be conducted without contamination concerns and reduction in set up time, making it ideal for near-field diagnosis. © 2017

  12. Numerical determination of lateral loss coefficients for subchannel analysis in nuclear fuel bundles

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sin Kim; Goon-Cherl Park

    1995-09-01

    An accurate prediction of cross-flow based on detailed knowledge of the velocity field in subchannels of a nuclear fuel assembly is of importance in nuclear fuel performance analysis. In this study, the low-Reynolds number {kappa}-{epsilon} turbulence model has been adopted in two adjacent subchannels with cross-flow. The secondary flow is estimated accurately by the anisotropic algebraic Reynolds stress model. This model was numerically calculated by the finite element method and has been verified successfully through comparison with existing experimental data. Finally, with the numerical analysis of the velocity field in such subchannel domain, an analytical correlation of the lateral lossmore » coefficient is obtained to predict the cross-flow rate in subchannel analysis codes. The correlation is expressed as a function of the ratio of the lateral flow velocity to the donor subchannel axial velocity, recipient channel Reynolds number and pitch-to-diameter.« less

  13. Distant touch hydrodynamic imaging with an artificial lateral line.

    PubMed

    Yang, Yingchen; Chen, Jack; Engel, Jonathan; Pandya, Saunvit; Chen, Nannan; Tucker, Craig; Coombs, Sheryl; Jones, Douglas L; Liu, Chang

    2006-12-12

    Nearly all underwater vehicles and surface ships today use sonar and vision for imaging and navigation. However, sonar and vision systems face various limitations, e.g., sonar blind zones, dark or murky environments, etc. Evolved over millions of years, fish use the lateral line, a distributed linear array of flow sensing organs, for underwater hydrodynamic imaging and information extraction. We demonstrate here a proof-of-concept artificial lateral line system. It enables a distant touch hydrodynamic imaging capability to critically augment sonar and vision systems. We show that the artificial lateral line can successfully perform dipole source localization and hydrodynamic wake detection. The development of the artificial lateral line is aimed at fundamentally enhancing human ability to detect, navigate, and survive in the underwater environment.

  14. Numerical and experimental studies on pulsatile flow in aneurysms arising laterally from a curved parent vessel at various angles.

    PubMed

    Liou, Tong-Miin; Li, Yi-Chen; Juan, Wei-Cheng

    2007-01-01

    Both numerical and experimental studies have been performed to characterize the fluid flow inside the lateral aneurysms arising from the curved parent vessels at various angles gamma. The implicit solver was based on the time-dependent Navier-Stokes equations of incompressible laminar flow. Solutions were generated by a cell-center finite-volume method that used second order upwind and second order center flux difference splitting for the convection and diffusion term, respectively. The second order Crank-Nicolson method was used in the time integration term while the SIMPLEC algorithm was adopted to handle the pressure-velocity coupling. Complementarily, the particle tracking velocimetry (PTV) was used to measure the velocity fields. The conditions selected were to simulate an internal carotid artery with a diameter of 5 mm by similarity rules. The values of gamma explored were 0 degrees, 45 degrees, 90 degrees, and 135 degrees. Pulsatile flow with Wormersley number 3.9 and Reynolds numbers varying from 350 to 850 was considered. The computed results are firstly verified by the PTV measured ones. Discussion of the results is in terms of pulsatile main and secondary velocity vector fields, inflow rates into the aneurysm, and the distributions of wall shear stress and static pressure. It is found that among the angles examined gamma=45( composite function) is the riskiest angle from a fluid dynamics point of view and the aneurysmal dome is at risk.

  15. Topics in Chemical Instrumentation--An Introduction to Supercritical Fluid Chromatography: Part 1: Principles and Instrumentation.

    ERIC Educational Resources Information Center

    Palmieri, Margo D.

    1988-01-01

    Identifies the properties and characteristics of supercritical fluids. Discusses the methodology for supercritical fluid chromatography including flow rate, plate height, column efficiency, viscosity, and other factors. Reviews instruments, column types, and elution conditions. Lists supercritical fluid data for 22 compounds, mostly organic. (MVL)

  16. Metal-organic frameworks in chromatography.

    PubMed

    Yusuf, Kareem; Aqel, Ahmad; ALOthman, Zeid

    2014-06-27

    Metal-organic frameworks (MOFs) emerged approximately two decades ago and are the youngest class of porous materials. Despite their short existence, MOFs are finding applications in a variety of fields because of their outstanding chemical and physical properties. This review article focuses on the applications of MOFs in chromatography, including high-performance liquid chromatography (HPLC), gas chromatography (GC), and other chromatographic techniques. The use of MOFs in chromatography has already had a significant impact; however, the utilisation of MOFs in chromatography is still less common than other applications, and the number of MOF materials explored in chromatography applications is limited. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Method for the quantification of vanadyl porphyrins in fractions of crude oils by High Performance Liquid Chromatography-Flow Injection-Inductively Coupled Plasma Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Wandekoken, Flávia G.; Duyck, Christiane B.; Fonseca, Teresa C. O.; Saint'Pierre, Tatiana D.

    2016-05-01

    High performance liquid chromatography hyphenated by flow injection to inductively coupled plasma mass spectrometry (HPLC-FI-ICP-MS) was used to investigate V linked to porphyrins present in fractions of crude oil. First, the crude oil sample was submitted to fractionation by preparative liquid chromatography with UV detection, at the porphyrin Soret band wavelength (400 nm). The obtained porphyrin fractions were then separated in a 250 mm single column, in the HPLC, and eluted with different mobile phases (methanol or methanol:toluene (80:20; v:v)). The quantification of V-porphyrins in the fractions eluted from HPLC was carried out by online measuring the 51V isotope in the ICP-MS, against vanadyl octaethylporphine standard solutions (VO-OEP), prepared in the same solvent as the mobile phase, and injected post-column directly into the plasma. A 20 μg L- 1 Ge in methanol was used as internal standard for minimizing non-spectral interference, such as short-term variations due to injection. The mathematical treatment of the signal based on Fast Fourier Transform smoothing algorithm was employed to improve the precision. The concentrations of V as V-porphyrins were between 2.7 and 11 mg kg- 1 in the fractions, which were close to the total concentration of V in the porphyrin fractions of the studied crude oil.

  18. Hydrocephalus secondary to obstruction of the lateral apertures in two dogs.

    PubMed

    Kent, M; Glass, E N; Haley, A C; Shaikh, L S; Sequel, M; Blas-Machado, U; Bishop, T M; Holmes, S P; Platt, S R

    2016-11-01

    Traditionally, hydrocephalus is divided into communicating or non-communicating (obstructive) based on the identification of a blockage of cerebrospinal fluid (CSF) flow through the ventricular system. Hydrocephalus ex vacuo refers to ventricular enlargement as a consequence of neuroparenchymal loss. Hydrocephalus related to obstruction of the lateral apertures of the fourth ventricles has rarely been described. The clinicopathologic findings in two dogs with hydrocephalus secondary to obstruction of the lateral apertures of the fourth ventricle are reported. Signs were associated with a caudal cervical spinal cord lesion in one dog and a caudal brain stem lesion in the other dog. Magnetic resonance imaging (MRI) disclosed dilation of the ventricular system, including the lateral recesses of the fourth ventricle. In one dog, postmortem ventriculography confirmed obstruction of the lateral apertures. Microscopic changes were identified in the choroid plexus in both dogs, yet a definitive cause of the obstructions was not identified. The MRI findings in both dogs are similar to membranous occlusion of the lateral and median apertures in human patients. MRI detection of dilation of the entire ventricular system in the absence of an identifiable cause should prompt consideration of an obstruction of the lateral apertures. In future cases, therapeutic interventions aimed at re-establishing CSF flow or ventriculoperitoneal catheterisation should be considered. © 2016 Australian Veterinary Association.

  19. Applicability of preparative overpressured layer chromatography and direct bioautography in search of antibacterial chamomile compounds.

    PubMed

    Móricz, Agnes M; Ott, Péter G; Alberti, Agnes; Böszörményi, Andrea; Lemberkovics, Eva; Szoke, Eva; Kéry, Agnes; Mincsovics, Emil

    2013-01-01

    In situ sample preparation and preparative overpressured layer chromatography (OPLC) fractionation on a 0.5 mm thick adsorbent layer of chamomile flower methanol extract prepurified by conventional gravitation accelerated column chromatography were applied in searching for bioactive components. Sample cleanup in situ on the adsorbent layer subsequent to sample application was performed using mobile phase flow in the opposite direction (the input and output of the eluent was exchanged). The antibacterial effect of the fractions obtained from the stepwise gradient OPLC separation with the flow in the normal direction was evaluated by direct bioautography against two Gram-negative bacteria: the luminescence gene tagged plant pathogenic Pseudomonas syringae pv. maculicola, and the naturally luminescent marine bacterium Vibrio fischeri. The fractions having strong activity were analyzed by SPME-GC/MS and HPLC/MS/MS. Mainly essential oil components, coumarins, flavonoids, phenolic acids, and fatty acids were tentatively identified in the fractions.

  20. Dual-Quantum-Dots-Labeled Lateral Flow Strip Rapidly Quantifies Procalcitonin and C-reactive Protein

    NASA Astrophysics Data System (ADS)

    Qi, XiaoPing; Huang, YunYe; Lin, ZhongShi; Xu, Liang; Yu, Hao

    2016-03-01

    In the article, a dual-quantum-dots-labeled (dual-QDs-labeled) lateral flow strip (LFS) method was developed for the simultaneous and rapid quantitative detection of procalcitonin (PCT) and C-reactive protein (CRP) in the blood. Two QD-antibody conjugates with different fluorescence emission spectra were produced and sprayed on the LFS to capture PCT and CRP in the blood. Furthermore, a double antibody sandwich method for PCT and, meanwhile, a competitive inhibition method for CRP were employed in the LFS. For PCT and CRP in serum assayed by the dual-QDs-labeled LFS, their detection sensitivities reached 0.1 and 1 ng/mL, respectively, and their linear quantitative detection ranges were from 0.3 to 200 ng/mL and from 50 to 250 μg/mL, respectively. There was little evidence that the PCT and CRP assays would be interfered with each other. The correlations for testing CRP and PCT in clinical samples were 99.75 and 97.02 %, respectively, between the dual-QDs-labeled LFS we developed and commercial methods. The rapid quantification of PCT and CRP on dual-QDs-labeled LFS is of great clinical value to distinguish inflammation, bacterial infection, or viral infection and to provide guidance for the use of antibiotics or other medicines.

  1. Gel compression considerations for chromatography scale-up for protein C purification.

    PubMed

    He, W; Bruley, D F; Drohan, W N

    1998-01-01

    This work is to establish theoretical and experimental relationships for the scale-up of Immobilized Metal Affinity Chromatography (IMAC) and Immuno Affinity Chromatography for the low cost production of large quantities of Protein C. The external customer requirements for this project have been established for Protein C deficient people with the goal of providing prophylactic patient treatment. Deep vein thrombosis is the major symptom for protein C deficiency creating the potential problem of embolism transport to important organs, such as, lung and brain. Gel matrices for protein C separation are being analyzed to determine the relationship between the material properties of the gel and the column collapse characteristics. The fluid flow rate and pressure drop is being examined to see how they influence column stability. Gel packing analysis includes two considerations; one is bulk compression due to flow rate, and the second is gel particle deformation due to fluid flow and pressure drop. Based on the assumption of creeping flow, Darcy's law is being applied to characterize the flow through the gel particles. Biot's mathematical description of three-dimensional consolidation in porous media is being used to develop a set of system equations. Finite difference methods are being utilized to obtain the equation solutions. In addition, special programs such as finite element approaches, ABAQUS, will be studied to determine their application to this particular problem. Experimental studies are being performed to determine flow rate and pressure drop correlation for the chromatographic columns with appropriate gels. Void fraction is being measured using pulse testing to allow Reynolds number calculations. Experimental yield stress is being measured to compare with the theoretical calculations. Total Quality Management (TQM) tools have been utilized to optimize this work. For instance, the "Scatter Diagram" has been used to evaluate and select the appropriate gels and

  2. Ecohydrologic Investigations of Shallow Lateral Subsurface Flow in Tropical Soils using Time-Lapse Surface Electrical Resistivity Tomography

    NASA Astrophysics Data System (ADS)

    Ogden, F. L.; Mojica, A.; Abebe, N. A.; Smithsonian Tropical Research Institute, Panama Canal Watershed Experiment, Agua Salud Project

    2010-12-01

    flow velocities over 1 m/h, presumably due to the existing downslope macroporosity network. These observations are being used to estimate macroporosity network properties and constrain hydrologic model parameters in different land uses. These results show that these non-invasive tests are a useful tool to determine the distribution of downslope lateral flow generated from pit and surface-applied saline solutions. ERT experimental results from a hillslope-scale experiment in central Panama, showing change in electrical conductivity from 30-minutes to 330-minutes after continuous injection of salinity contrast at x=0.

  3. Evaluation of a direct injection nebulizer interface for flow injection analysis and high performance liquid chromatography with inductively coupled plasma-atomic emission spectroscopic detection

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    LaFreniere, K.E.

    A direct injection nebulizer (DIN) was designed, developed and evaluated to determine its potential utilization as an effective interface for flow injection analysis (FIA) and high performance liquid chromatography (HPLC) coupled with inductively coupled plasma-atomic emission spectroscopic detection. The analytical figures of merit for the DIN when used as an interface for FIA-ICP-AES were found to be comparable to or better than those obtained with conventional pneumatic nebulization in terms of limits of detection (LODs), reproducibility, linearity, and interelement effects. In the HPLC mode, the LODDs were found to be comparable to those obtained by continuous-flow sample introduction into themore » ICP, or inferior by up to only a factor of four. Stable plasma operation was maintained for the DIN sample introduction of a variety of pure organic solvents, including acetonitrile, methanol, methyl-isobutylketone, and pyridine. The HPLC-DIN-ICP-AES facility was specifically applied for the speciation of inorganic and organo-metallic species contained in synthetic mixtures, vanilla extracts and a variety of energy-related materials, such as shale oil process water, coal extracts, shale oil, crude oil, and an SRC II. Suggestions for future research are also considered.« less

  4. Transfer of a three step mAb chromatography process from batch to continuous: Optimizing productivity to minimize consumable requirements.

    PubMed

    Gjoka, Xhorxhi; Gantier, Rene; Schofield, Mark

    2017-01-20

    The goal of this study was to adapt a batch mAb purification chromatography platform for continuous operation. The experiments and rationale used to convert from batch to continuous operation are described. Experimental data was used to design chromatography methods for continuous operation that would exceed the threshold for critical quality attributes and minimize the consumables required as compared to batch mode of operation. Four unit operations comprising of Protein A capture, viral inactivation, flow-through anion exchange (AEX), and mixed-mode cation exchange chromatography (MMCEX) were integrated across two Cadence BioSMB PD multi-column chromatography systems in order to process a 25L volume of harvested cell culture fluid (HCCF) in less than 12h. Transfer from batch to continuous resulted in an increase in productivity of the Protein A step from 13 to 50g/L/h and of the MMCEX step from 10 to 60g/L/h with no impact on the purification process performance in term of contaminant removal (4.5 log reduction of host cell proteins, 50% reduction in soluble product aggregates) and overall chromatography process yield of recovery (75%). The increase in productivity, combined with continuous operation, reduced the resin volume required for Protein A and MMCEX chromatography by more than 95% compared to batch. The volume of AEX membrane required for flow through operation was reduced by 74%. Moreover, the continuous process required 44% less buffer than an equivalent batch process. This significant reduction in consumables enables cost-effective, disposable, single-use manufacturing. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  5. Improved purification of immunoglobulin G from plasma by mixed-mode chromatography.

    PubMed

    Chai, Dong-Sheng; Sun, Yan; Wang, Xiao-Ning; Shi, Qing-Hong

    2014-12-01

    Efficient loading of immunoglobulin G in mixed-mode chromatography is often a serious bottleneck in the chromatographic purification of immunoglobulin G. In this work, a mixed-mode ligand, 4-(1H-imidazol-1-yl) aniline, was coupled to Sepharose Fast Flow to fabricate AN SepFF adsorbents with ligand densities of 15-64 mmol/L, and the chromatographic performances of these adsorbents were thoroughly investigated to identify a feasible approach to improve immunoglobulin G purification. The results indicate that a critical ligand density exists for immunoglobulin G on the AN SepFF adsorbents. Above the critical ligand density, the adsorbents showed superior selectivity to immunoglobulin G at high salt concentrations, and also exhibited much higher dynamic binding capacities. For immunoglobulin G purification, both the yield and binding capacity increased with adsorbent ligand density along with a decrease in purity. It is difficult to improve the binding capacity, purity, and yield of immunoglobulin G simultaneously in AN SepFF chromatography. By using tandem AN SepFF chromatography, a threefold increase in binding capacity as well as high purity and yield of immunoglobulin G were achieved. Therefore, the tandem chromatography demonstrates that AN SepFF adsorbent is a practical and feasible alternative to MEP HyperCel adsorbents for immunoglobulin G purification. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Lateralization of cerebral blood flow velocity changes during auditory stimulation: a functional transcranial Doppler study.

    PubMed

    Carod Artal, Francisco Javier; Vázquez Cabrera, Carolina; Horan, Thomas Anthony

    2004-01-01

    Transcranial Doppler ultrasonography (TCD) permits the assessment of cognitively induced cerebral blood flow velocity (BFV) changes. We sought to investigate the lateralization of BFV acceleration induced by auditory stimulation and speech in a normal population. TCD monitoring of BFV in the middle cerebral arteries (MCA) was performed in 30 normal right-handed volunteers (average age = 31.7 years). Noise stimulation, speech, and instrumental music were administered during 60 sec to both ears by means of earphones. Auditory stimulation induced a significant BFV increase in the ipsilateral MCA compared to BFV during the preceding rest periods. Left MCA BFV increased by an average of 7.1% (noise), 8.4% (language), and 5.2% (melody) over baseline values, and right MCA BFV increased 5.1%, 3.1%, and 4.2%, respectively. Speech stimulation produced a significant increase in BFV in the left hemisphere MCA (from 49.86 to 54.03 cm/sec; p < .0001). Left MCA BFV response to speech stimulation may reflect the dominance of the left hemisphere in language processing by right-handed individuals. Due to the high temporal resolution of TCD we were able show a habituation effect during the 60-sec stimulation period.

  7. A Novel Isothermal Assay of Borrelia burgdorferi by Recombinase Polymerase Amplification with Lateral Flow Detection.

    PubMed

    Liu, Wei; Liu, Hui-Xin; Zhang, Lin; Hou, Xue-Xia; Wan, Kang-Lin; Hao, Qin

    2016-08-03

    A novel isothermal detection for recombinase polymerase amplification with lateral flow (LF-RPA) was established for Borrelia burgdorferi (B. burgdorferi) detection in this study. This assay with high sensitivity and specificity can get a visible result without any additional equipment in 30 min. We designed a pair of primers according to recA gene of B. burgdorferi strains and a methodology evaluation was performed. The results showed that the RPA assay based on the recA gene was successfully applied in B. burgdorferi detection, and its specific amplification was only achieved from the genomic DNA of B. burgdorferi. The detection limit of the new assay was about 25 copies of the B. burgdorferi genomic DNA. Twenty Lyme borreliosis patients' serum samples were detected by LF-RPA assay, real-time qPCR and nested-PCR. Results showed the LF-RPA assay is more effective than nested-PCR for its shorter reaction time and considerably higher detection rate. This method is of great value in clinical rapid detection for Lyme borreliosis. Using the RPA assay might be a megatrend for DNA detection in clinics and endemic regions.

  8. Column Liquid Chromatography.

    ERIC Educational Resources Information Center

    Majors, Ronald E.; And Others

    1984-01-01

    Reviews literature covering developments of column liquid chromatography during 1982-83. Areas considered include: books and reviews; general theory; columns; instrumentation; detectors; automation and data handling; multidimensional chromatographic and column switching techniques; liquid-solid chromatography; normal bonded-phase, reversed-phase,…

  9. Rolling with the flow: bumblebees flying in unsteady wakes.

    PubMed

    Ravi, Sridhar; Crall, James D; Fisher, Alex; Combes, Stacey A

    2013-11-15

    Our understanding of how variable wind in natural environments affects flying insects is limited because most studies of insect flight are conducted in either smooth flow or still air conditions. Here, we investigate the effects of structured, unsteady flow (the von Karman vortex street behind a cylinder) on the flight performance of bumblebees (Bombus impatiens). Bumblebees are 'all-weather' foragers and thus frequently experience variable aerial conditions, ranging from fully mixed, turbulent flow to unsteady, structured vortices near objects such as branches and stems. We examined how bumblebee flight performance differs in unsteady versus smooth flow, as well as how the orientation of unsteady flow structures affects their flight performance, by filming bumblebees flying in a wind tunnel under various flow conditions. The three-dimensional flight trajectories and orientations of bumblebees were quantified in each of three flow conditions: (1) smooth flow, (2) the unsteady wake of a vertical cylinder (inducing strong lateral disturbances) and (3) the unsteady wake of a horizontal cylinder (inducing strong vertical disturbances). In both unsteady conditions, bumblebees attenuated the disturbances induced by the wind quite effectively, but still experienced significant translational and rotational fluctuations as compared with flight in smooth flow. Bees appeared to be most sensitive to disturbance along the lateral axis, displaying large lateral accelerations, translations and rolling motions in response to both unsteady flow conditions, regardless of orientation. Bees also displayed the greatest agility around the roll axis, initiating voluntary casting maneuvers and correcting for lateral disturbances mainly through roll in all flow conditions. Both unsteady flow conditions reduced the upstream flight speed of bees, suggesting an increased cost of flight in unsteady flow, with potential implications for foraging patterns and colony energetics in natural

  10. Evaluation of IDA-PEVA hollow fiber membrane metal ion affinity chromatography for purification of a histidine-tagged human proinsulin.

    PubMed

    de Aquino, Luciana Cristina Lins; de Sousa, Heloisa Ribeiro Tunes; Miranda, Everson Alves; Vilela, Luciano; Bueno, Sônia Maria Alves

    2006-04-13

    Inabilities to process particulate material and to allow the use of high flow rates are limitations of conventional chromatography. Membranes have been suggested as matrix for affinity separation due to advantages such as allowing high flow rates and low-pressure drops. This work evaluated the feasibility of using an iminodiacetic acid linked poly(ethylenevinyl alcohol) membrane in the immobilized metal ion affinity chromatography (IMAC) purification of a human proinsulin(His)(6) of an industrial insulin production process. The screening of metal ions showed Ni(2+) as metal with higher selectivity and capacity among the Cu(2+), Ni(2+), Zn(2+) and Co(2+). The membrane showed to be equivalent to conventional chelating beads in terms of selectivity and had a lower capacity (3.68 mg/g versus 12.26 mg/g). The dynamic adsorption capacity for human proinsulin(His)(6) was unaffected by the mode of operation (dead-end and cross-flow filtration).

  11. Lateral Flow Immunoassays for Ebola Virus Disease Detection in Liberia.

    PubMed

    Phan, Jill C; Pettitt, James; George, Josiah S; Fakoli, Lawrence S; Taweh, Fahn M; Bateman, Stacey L; Bennett, Richard S; Norris, Sarah L; Spinnler, David A; Pimentel, Guillermo; Sahr, Phillip K; Bolay, Fatorma K; Schoepp, Randal J

    2016-10-15

    Lateral flow immunoassays (LFIs) are point-of-care diagnostic assays that are designed for single use outside a formal laboratory, with in-home pregnancy tests the best-known example of these tests. Although the LFI has some limitations over more-complex immunoassay procedures, such as reduced sensitivity and the potential for false-positive results when using complex sample matrices, the assay has the benefits of a rapid time to result and ease of use. These benefits make it an attractive option for obtaining rapid results in an austere environment. In an outbreak of any magnitude, a field-based rapid diagnostic assay would allow proper patient transport and for safe burials to be conducted without the delay caused by transport of samples between remote villages and testing facilities. Use of such point-of-care instruments in the ongoing Ebola virus disease (EVD) outbreak in West Africa would have distinct advantages in control and prevention of local outbreaks, but proper understanding of the technology and interpretation of results are important. In this study, a LFI, originally developed by the Naval Medical Research Center for Ebola virus environmental testing, was evaluated for its ability to detect the virus in clinical samples in Liberia. Clinical blood and plasma samples and post mortem oral swabs submitted to the Liberian Institute for Biomedical Research, the National Public Health Reference Laboratory for EVD testing, were tested and compared to results of real-time reverse transcription-polymerase chain reaction (rRT-PCR), using assays targeting Ebola virus glycoprotein and nucleoprotein. The LFI findings correlated well with those of the real-time RT-PCR assays used as benchmarks. Rapid antigen-detection tests such as LFIs are attractive alternatives to traditional immunoassays but have reduced sensitivity and specificity, resulting in increases in false-positive and false-negative results. An understanding of the strengths, weaknesses, and

  12. Kool-Aid Chromatography

    ERIC Educational Resources Information Center

    Jenkins, Christie L.

    1986-01-01

    Offers guidelines and suggests activities that can introduce middle school students to the process and principles of chromatography in an inexpensive and safe manner. Proposes that experiences with Kool-aid and food coloring chromatography can provide insights into how scientists think, work, and communicate. (ML)

  13. Hydrocode Analysis of Lateral Stress Gauges in Shocked Tantalum

    NASA Astrophysics Data System (ADS)

    Harris, Ernest; Winter, Ron

    2007-06-01

    Experiements published by other workers on the resistance change of manganin stress gauges embedded in a lateral orientation in Tantalum targets have been analysed using an Adaptive Mesh Refinement Hydrocode. It was found that for four experiments the shape of the time profile of the computed lateral stress in the mounting layer closely matched the shape of the experimental lateral stress profiles. However, the calculated lateral stresses at the gauge location in the mounting layer are significantly less than the stresses that would have been produced in the target if no gauge had been present. The perturbation caused by the gauge increased as the strength of the applied shock increased. When the perturbations are taken into account values of flow stress that are significantly smaller than those reported in the original research paper are derived. The work demonstrates that the lateral gauge technique can give valuable information on strength provided high resolution simulation is used to compensate for the perturbations caused by the gauges.

  14. Fiber-based monolithic columns for liquid chromatography.

    PubMed

    Ladisch, Michael; Zhang, Leyu

    2016-10-01

    Fiber-based monoliths for use in liquid chromatographic separations are defined by columns packed with aligned fibers, woven matrices, or contiguous fiber structures capable of achieving rapid separations of proteins, macromolecules, and low molecular weight components. A common denominator and motivating driver for this approach, first initiated 25 years ago, was reducing the cost of bioseparations in a manner that also reduced residence time of retained components while achieving a high ratio of mass to momentum transfer. This type of medium, when packed into a liquid chromatography column, minimized the fraction of stagnant liquid and resulted in a constant plate height for non-adsorbing species. The uncoupling of dispersion from eluent flow rate enabled the surface chemistry of the stationary phase to be considered separately from fluid transport phenomena and pointed to new ways to apply chemistry for the engineering of rapid bioseparations. This paper addresses developments and current research on fiber-based monoliths and explains how the various forms of this type of chromatographic stationary phase have potential to provide new tools for analytical and preparative scale separations. The different stationary phases are discussed, and a model that captures the observed constant plate height as a function of mobile phase velocity is reviewed. Methods that enable hydrodynamically stable fiber columns to be packed and operated over a range of mobile phase flow rates, together with the development of new fiber chemistries, are shown to provide columns that extend the versatility of liquid chromatography using monoliths, particularly at the preparative scale. Graphical Abstract Schematic representation of a sample mixture being separated by a rolled-stationary phase column, resulting separated peaks shown in the chromatogram.

  15. Earthflow yield strength constrained by lateral levee morphology

    NASA Astrophysics Data System (ADS)

    Nereson, A. L.; Finnegan, N. J.

    2015-12-01

    Slow-moving landslides, or earthflows, are characterized by persistent, flow-like motion that is commonly modeled using various viscous and viscoplastic rheologies. One of the manifestations of viscoplastic flow down a slope is the emergence of stationary bodies of fluid at the margins of the flow (i.e. lateral levees). These levees are common signatures of earthflow morphology and, while they are frequently used to outline boundaries for mapping purposes, they have received little attention for what they may indicate about the history and properties of the flow itself. In contrast, lateral levees along lava flows have long been used by physical volcanologists as tools to learn about their non-Newtonian rheologies and chemical compositions. Hulme (1974) was the first to note that, for a given slope, levee width may be characteristic of a fluids's yield strength and his methodology has been subsequently used to infer properties of lavas on the Earth, the Moon, and Mars. Using these lavas as analogies, we apply Hulme's approach to earthflows in a variety of settings globally. We find that calculated yield strengths for individual earthflows fall within a relatively narrow range between 101-102 kPa. In addition, individual earthflow complexes often preserve multiple generations of levees, which in some cases may record apparent reductions in yield strength over time for a given flow, possibly from weakening of previously failed material. Knowledge of earthflow yield strength permits the calculation of a critical earthflow thickness below which there will be no downslope motion for a given slope angle. Thicknesses calculated in this manner could thus be used to estimate the flux of landslide material for earthflows without direct depth constraints, provided that surface velocity measurements are obtained by other methods (e.g. InSAR, GPS, manual feature tracking).

  16. Hydrocode Analysis of Lateral Stress Gauges in Shocked Tantalum

    NASA Astrophysics Data System (ADS)

    Harris, E. J.; Winter, R. E.

    2007-12-01

    Experiments published by other workers, on the resistance change of manganin stress gauges embedded in a lateral orientation in tantalum targets shocked to a range of stresses, have been analysed using an adaptive mesh refinement hydrocode. It was found that for all of the four experiments the shape of the time profile of the computed lateral stress in the mounting layer closely matched the shape of the experimental lateral stress profiles. However, the calculated lateral stresses at the gauge location in the mounting layer are significantly less than the lateral stresses that would have been produced in the target if no gauge had been present. The perturbation caused by the gauge increased as the strength of the applied shock increased. When the perturbations are taken into account values of flow stress that are significantly smaller than those reported in the original research paper are derived. The work shows that the lateral gauge technique can give valuable information on strength provided high resolution simulation is used to compensate for the perturbations caused by the gauges.

  17. A detached eddy simulation model for the study of lateral separation zones along a large canyon-bound river

    NASA Astrophysics Data System (ADS)

    Alvarez, Laura V.; Schmeeckle, Mark W.; Grams, Paul E.

    2017-01-01

    Lateral flow separation occurs in rivers where banks exhibit strong curvature. In canyon-bound rivers, lateral recirculation zones are the principal storage of fine-sediment deposits. A parallelized, three-dimensional, turbulence-resolving model was developed to study the flow structures along lateral separation zones located in two pools along the Colorado River in Marble Canyon. The model employs the detached eddy simulation (DES) technique, which resolves turbulence structures larger than the grid spacing in the interior of the flow. The DES-3D model is validated using Acoustic Doppler Current Profiler flow measurements taken during the 2008 controlled flood release from Glen Canyon Dam. A point-to-point validation using a number of skill metrics, often employed in hydrological research, is proposed here for fluvial modeling. The validation results show predictive capabilities of the DES model. The model reproduces the pattern and magnitude of the velocity in the lateral recirculation zone, including the size and position of the primary and secondary eddy cells, and return current. The lateral recirculation zone is open, having continuous import of fluid upstream of the point of reattachment and export by the recirculation return current downstream of the point of separation. Differences in magnitude and direction of near-bed and near-surface velocity vectors are found, resulting in an inward vertical spiral. Interaction between the recirculation return current and the main flow is dynamic, with large temporal changes in flow direction and magnitude. Turbulence structures with a predominately vertical axis of vorticity are observed in the shear layer becoming three-dimensional without preferred orientation downstream.

  18. Predicting Peak Flows following Forest Fires

    NASA Astrophysics Data System (ADS)

    Elliot, William J.; Miller, Mary Ellen; Dobre, Mariana

    2016-04-01

    Following forest fires, peak flows in perennial and ephemeral streams often increase by a factor of 10 or more. This increase in peak flow rate may overwhelm existing downstream structures, such as road culverts, causing serious damage to road fills at stream crossings. In order to predict peak flow rates following wildfires, we have applied two different tools. One is based on the U.S.D.A Natural Resource Conservation Service Curve Number Method (CN), and the other is by applying the Water Erosion Prediction Project (WEPP) to the watershed. In our presentation, we will describe the science behind the two methods, and present the main variables for each model. We will then provide an example of a comparison of the two methods to a fire-prone watershed upstream of the City of Flagstaff, Arizona, USA, where a fire spread model was applied for current fuel loads, and for likely fuel loads following a fuel reduction treatment. When applying the curve number method, determining the time to peak flow can be problematic for low severity fires because the runoff flow paths are both surface and through shallow lateral flow. The WEPP watershed version incorporates shallow lateral flow into stream channels. However, the version of the WEPP model that was used for this study did not have channel routing capabilities, but rather relied on regression relationships to estimate peak flows from individual hillslope polygon peak runoff rates. We found that the two methods gave similar results if applied correctly, with the WEPP predictions somewhat greater than the CN predictions. Later releases of the WEPP model have incorporated alternative methods for routing peak flows that need to be evaluated.

  19. Function Lateralization via Measuring Coherence Laterality

    PubMed Central

    Wang, Ze; Mechanic-Hamilton, Dawn; Pluta, John; Glynn, Simon; Detre, John A.

    2009-01-01

    A data-driven approach for lateralization of brain function based on the spatial coherence difference of functional MRI (fMRI) data in homologous regions-of-interest (ROI) in each hemisphere is proposed. The utility of using coherence laterality (CL) to determine function laterality was assessed first by examining motor laterality using normal subjects’ data acquired both at rest and with a simple unilateral motor task and subsequently by examining mesial temporal lobe memory laterality in normal subjects and patients with temporal lobe epilepsy. The motor task was used to demonstrate that CL within motor ROI correctly lateralized functional stimulation. In patients with unilateral epilepsy studied during a scene-encoding task, CL in a hippocampus-parahippocampus-fusiform (HPF) ROI was concordant with lateralization based on task activation, and the CL index (CLI) significantly differentiated the right side group to the left side group. By contrast, normal controls showed a symmetric HPF CLI distribution. Additionally, similar memory laterality prediction results were still observed using CL in epilepsy patients with unilateral seizures after the memory encoding effect was removed from the data, suggesting the potential for lateralization of pathological brain function based on resting fMRI data. A better lateralization was further achieved via a combination of the proposed approach and the standard activation based approach, demonstrating that assessment of spatial coherence changes provides a complementary approach to quantifying task-correlated activity for lateralizing brain function. PMID:19345736

  20. Turbulent flow chromatography TFC-tandem mass spectrometry supporting in vitro/vivo studies of NCEs in high throughput fashion.

    PubMed

    Verdirame, Maria; Veneziano, Maria; Alfieri, Anna; Di Marco, Annalise; Monteagudo, Edith; Bonelli, Fabio

    2010-03-11

    Turbulent Flow Chromatography (TFC) is a powerful approach for on-line extraction in bioanalytical studies. It improves sensitivity and reduces sample preparation time, two factors that are of primary importance in drug discovery. In this paper the application of the ARIA system to the analytical support of in vivo pharmacokinetics (PK) and in vitro drug metabolism studies is described, with an emphasis in high throughput optimization. For PK studies, a comparison between acetonitrile plasma protein precipitation (APPP) and TFC was carried out. Our optimized TFC methodology gave better S/N ratios and lower limit of quantification (LOQ) than conventional procedures. A robust and high throughput analytical method to support hepatocyte metabolic stability screening of new chemical entities was developed by hyphenation of TFC with mass spectrometry. An in-loop dilution injection procedure was implemented to overcome one of the main issues when using TFC, that is the early elution of hydrophilic compounds that renders low recoveries. A comparison between off-line solid phase extraction (SPE) and TFC was also carried out, and recovery, sensitivity (LOQ), matrix effect and robustness were evaluated. The use of two parallel columns in the configuration of the system provided a further increase of the throughput. Copyright 2009 Elsevier B.V. All rights reserved.

  1. Effects of salts on protein-surface interactions: applications for column chromatography.

    PubMed

    Tsumoto, Kouhei; Ejima, Daisuke; Senczuk, Anna M; Kita, Yoshiko; Arakawa, Tsutomu

    2007-07-01

    Development of protein pharmaceuticals depends on the availability of high quality proteins. Various column chromatographies are used to purify proteins and characterize the purity and properties of the proteins. Most column chromatographies require salts, whether inorganic or organic, for binding, elution or simply better recovery and resolution. The salts modulate affinity of the proteins for particular columns and nonspecific protein-protein or protein-surface interactions, depending on the type and concentration of the salts, in both specific and nonspecific manners. Salts also affect the binding capacity of the column, which determines the size of the column to be used. Binding capacity, whether equilibrium or dynamic (under an approximation of a slow flow rate), depends on the binding constant, protein concentration and the number of the binding site on the column as well as nonspecific binding. This review attempts to summarize the mechanism of the salt effects on binding affinity and capacity for various column chromatographies and on nonspecific protein-protein or protein-surface interactions. Understanding such salt effects should also be useful in preventing nonspecific protein binding to various containers. Copyright 2007 Wiley-Liss, Inc.

  2. Variability of Rheotaxis Behaviors in Larval Bullfrogs Highlights Species Diversity in Lateral Line Function

    PubMed Central

    Brown, Erika E. A.

    2016-01-01

    The morphology and distribution of lateral line neuromasts vary between ecomorphological types of anuran tadpoles, but little is known about how this structural variability contributes to differences in lateral-line mediated behaviors. Previous research identified distinct differences in one such behavior, positive rheotaxis towards the source of a flow, in two tadpole species, the African clawed frog (Xenopus laevis; type 1) and the American bullfrog (Rana catesbeiana; type 4). Because these two species had been tested under different flow conditions, we re-evaluated these findings by quantifying flow-sensing behaviors of bullfrog tadpoles in the same flow field in which X. laevis tadpoles had been tested previously. Early larval bullfrog tadpoles were exposed to flow in the dark, in the presence of a discrete light cue, and after treatment with the ototoxin gentamicin. In response to flow, tadpoles moved downstream, closer to a side wall, and higher in the water column, but they did not station-hold. Tadpoles exhibited positive rheotaxis, but with long latencies, low to moderate accuracy, and considerable individual variability. This is in contrast to the robust, stereotyped station-holding and accurate rheotaxis of X. laevis tadpoles. The presence of a discrete visual cue and gentamicin treatment altered spatial positioning and disrupted rheotaxis in both tadpole species. Species differences in lateral-line mediated behaviors may reflect differences in neuromast number and distribution, life history, or perceptual salience of other environmental cues. PMID:27870909

  3. Reverse Phase-ultra Flow Liquid Chromatography-diode Array Detector Quantification of Anticancerous and Antidiabetic Drug Mangiferin from 11 Species of Swertia from India.

    PubMed

    Kshirsagar, Parthraj R; Gaikwad, Nikhil B; Panda, Subhasis; Hegde, Harsha V; Pai, Sandeep R

    2016-01-01

    Genus Swertia is valued for its great medicinal potential, mainly Swertia chirayita (Roxb. ex Fleming) H. Karst. is used in traditional medicine for a wide range of diseases. Mangiferin one of xanthoids is referred with enormous pharmacological potentials. The aim of the study was to quantify and compare the anticancerous and antidiabetic drug mangiferin from 11 Swertia species from India. The study also evaluates hierarchical relationships between the species based on mangiferin content using multivariate analysis. The reverse phase-ultra flow liquid chromatography-diode array detector analyses was performed and chromatographic separation was achieved on a Lichrospher 100, C18e (5 μm) column (250-4.6 mm). Mobile phase consisting of 0.2% triethylamine (pH-4 with O-phosphoric acid) and acetonitrile (85:15) was used for separation with injection volume 20 μL and detection wave length at 257 nm. Results indicated that concentration of mangiferin has been found to vary largely between Swertia species collected from different regions. Content of mangiferin was found to be highest in Swertia minor compared to other Swertia species studied herein from the Western Ghats and Himalayan region also. The same was also evident in the multivariate analysis, wherein S. chirayita, S. minor and Swertia paniculata made a separate clade. Conclusively, the work herein provides insights of mangiferin content from 11 Swertia species of India and also presents their hierarchical relationships. To best of the knowledge this is the first report of higher content of mangiferin from any Swertia species. The present study quantifies and compares mangiferin in 11 species of Swertia from India. The study also evaluates hierarchical relationships between the species based on mangiferin content using multivariate analysis. The mangiferin content was highest in S. minor compared to the studied Swertia species. To the best of our knowledge this is the first report of higher content of mangiferin

  4. Direct analysis of prostaglandin-E2 and -D2 produced in an inflammatory cell reaction and its application for activity screening and potency evaluation using turbulent flow chromatography liquid chromatography-high resolution mass spectrometry.

    PubMed

    Shin, Jeong-Sook; Peng, Lei; Kang, Kyungsu; Choi, Yongsoo

    2016-09-09

    Direct analysis of prostaglandin-E2 (PGE2) and -D2 (PGD2) produced from a RAW264.7 cell-based reaction was performed by liquid chromatography high-resolution mass spectrometry (LC-HRMS), which was online coupled with turbulent flow chromatography (TFC). The capability of this method to accurately measure PG levels in cell reaction medium containing cytokines or proteins as a reaction byproduct was cross-validated by two conventional methods. Two methods, including an LC-HRMS method after liquid-liquid extraction (LLE) of the sample and a commercial PGE2 enzyme-linked immunosorbent assay (ELISA), showed PGE2 and/or PGD2 levels almost similar to those obtained by TFC LC-HRMS over the reaction time after LPS stimulation. After the cross-validation, significant analytical throughputs, allowing simultaneous screening and potency evaluation of 80 natural products including 60 phytochemicals and 20 natural product extracts for the inhibition of the PGD2 produced in the cell-based inflammatory reaction, were achieved using the TFC LC-HRMS method developed. Among the 60 phytochemicals screened, licochalcone A and formononetin inhibited PGD2 production the most with IC50 values of 126 and 151nM, respectively. For a reference activity, indomethacin and diclofenac were used, measuring IC50 values of 0.64 and 0.21nM, respectively. This method also found a butanol extract of Akebia quinata Decne (AQ) stem as a promising natural product for PGD2 inhibition. Direct and accurate analysis of PGs in the inflammatory cell reaction using the TFC LC-HRMS method developed enables the high-throughput screening and potency evaluation of as many as 320 samples in less than 48h without changing a TFC column. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. High Performance Thin Layer Chromatography.

    ERIC Educational Resources Information Center

    Costanzo, Samuel J.

    1984-01-01

    Clarifies where in the scheme of modern chromatography high performance thin layer chromatography (TLC) fits and why in some situations it is a viable alternative to gas and high performance liquid chromatography. New TLC plates, sample applications, plate development, and instrumental techniques are considered. (JN)

  6. Isolation of Methoxyfuranocoumarins From Ammi majus by Centrifugal Partition Chromatography.

    PubMed

    Bartnik, Magdalena; Mazurek, Anna Katarzyna

    2016-01-01

    Pure methoxyfuranocoumarins were isolated from Ammi majus L. by use of low-pressure column chromatography (LPCC) followed by centrifugal partition chromatography (CPC). The concentrated petroleum ether extract from fruits of A. majus was fractionated on a silica gel column using a gradient of ethyl acetate in dichloromethane (0-80%, v/v). Coumarin-rich fractions were analyzed by thin-layer chromatography (TLC) and high-performance liquid chromatography with diode array detection (HPLC/DAD). Xanthotoxin (8-MOP) and isopimpinellin (isoP), structurally similar compounds, were isolated in one fraction (FR6). To avoid multistep and long-lasting TLC preparation, optimization of CPC conditions has been performed. In one run, an effective separation of 8-MOP and isoP was achieved. The two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (10 : 8 : 10 : 9; v/v) in an ascending mode (the aqueous phase was a stationary phase, and the organic phase was a mobile phase), with flow rate 3 mL/min and rotation speed 1,600 r.p.m., was used. The identification and high purities of isolated 8-MOP (98.7%) and isoP (100%) were confirmed by HPLC/DAD assay, when compared with standards. The developed CPC method could be applied to the effective isolation of 8-MOP and isoP from plant extracts. The high purity of obtained compounds makes possible further exploitation of these components in biological studies. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. Three dimensional liquid chromatography coupling ion exchange chromatography/hydrophobic interaction chromatography/reverse phase chromatography for effective protein separation in top-down proteomics.

    PubMed

    Valeja, Santosh G; Xiu, Lichen; Gregorich, Zachery R; Guner, Huseyin; Jin, Song; Ge, Ying

    2015-01-01

    To address the complexity of the proteome in mass spectrometry (MS)-based top-down proteomics, multidimensional liquid chromatography (MDLC) strategies that can effectively separate proteins with high resolution and automation are highly desirable. Although various MDLC methods that can effectively separate peptides from protein digests exist, very few MDLC strategies, primarily consisting of 2DLC, are available for intact protein separation, which is insufficient to address the complexity of the proteome. We recently demonstrated that hydrophobic interaction chromatography (HIC) utilizing a MS-compatible salt can provide high resolution separation of intact proteins for top-down proteomics. Herein, we have developed a novel 3DLC strategy by coupling HIC with ion exchange chromatography (IEC) and reverse phase chromatography (RPC) for intact protein separation. We demonstrated that a 3D (IEC-HIC-RPC) approach greatly outperformed the conventional 2D IEC-RPC approach. For the same IEC fraction (out of 35 fractions) from a crude HEK 293 cell lysate, a total of 640 proteins were identified in the 3D approach (corresponding to 201 nonredundant proteins) as compared to 47 in the 2D approach, whereas simply prolonging the gradients in RPC in the 2D approach only led to minimal improvement in protein separation and identifications. Therefore, this novel 3DLC method has great potential for effective separation of intact proteins to achieve deep proteome coverage in top-down proteomics.

  8. Lateral weathering gradients in glaciated catchments

    NASA Astrophysics Data System (ADS)

    McGuire, K. J.; Bailey, S. W.; Ross, D. S.; Strahm, B. D.; Schreiber, M. E.

    2016-12-01

    Mineral dissolution and the distribution of weathering products are fundamental processes that drive development and habitability of the Earth's critical zone; yet, the spatial configuration of these processes in some systems is not well understood. Feedbacks between hydrologic flows and weathering fluxes are necessary to understanding how the critical zone develops. In upland glaciated catchments of the northeastern USA, primary mineral dissolution and the distribution of weathering products are spatially distinct and predictable over short distances. Hillslopes, where shallow soils force lateral hydrologic fluxes through accumulated organic matter, produce downslope gradients in mineral depletion, weathering product accumulation, soil development, and solute chemistry. We propose that linked gradients in hydrologic flow paths, soil depth, and vegetation lead to predictable differences in the location and extent of mineral dissolution in regolith (soil, subsoil, and rock fragments) and bedrock, and that headwater catchments within the upland glaciated northeast show a common architecture across hillslopes as a result. Examples of these patterns and processes will be illustrated using observations from the Hubbard Brook Experimental Forest in New Hampshire where laterally distinct soils with strong morphological and biogeochemical gradients have been documented. Patterns in mineral depletion and product accumulation are essential in predicting how ecosystems will respond to stresses, disturbance, and management.

  9. Protein separation through preliminary experiments concerning pH and salt concentration by tube radial distribution chromatography based on phase separation multiphase flow using a polytetrafluoroethylene capillary tube.

    PubMed

    Kan, Hyo; Tsukagoshi, Kazuhiko

    2017-07-01

    Protein mixtures were separated using tube radial distribution chromatography (TRDC) in a polytetrafluoroethylene (PTFE) capillary (internal diameter=100µm) separation tube. Separation by TRDC is based on the annular flow in phase separation multiphase flow and features an open-tube capillary without the use of specific packing agents or application of high voltages. Preliminary experiments were conducted to examine the effects of pH and salt concentration on the phase diagram of the ternary mixed solvent solution of water-acetonitrile-ethyl acetate (8:2:1 volume ratio) and on the TRDC system using the ternary mixed solvent solution. A model protein mixture containing peroxidase, lysozyme, and bovine serum albumin was analyzed via TRDC with the ternary mixed solvent solution at various pH values, i.e., buffer-acetonitrile-ethyl acetate (8:2:1 volume ratio). Protein was separated on the chromatograms by the TRDC system, where the elution order was determined by the relation between the isoelectric points of protein and the pH values of the solvent solution. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Rapid diagnosis of tuberculosis in dromedary camel (Camelus dromedarius) using lateral flow assay-based kit.

    PubMed

    Ranjan, Rakesh; Narnaware, Shirish D; Nath, Kashi; Sawal, R K; Patil, N V

    2018-04-01

    Accurate early antemortem diagnosis of tuberculosis in dromedary camels is difficult due to the lack of reliable diagnostic test. The present study aimed to evaluate a lateral flow assay-based kit (rapid assay kit) in tuberculosis diagnosis that employs immuno-chromatographic detection of antibodies in serum, plasma, or whole blood. In a dromedary camel herd comprising 337 animals located at Bikaner, Rajasthan, India, 50 adult weak camels (11 males and 39 females) were tested by applying a single intradermal tuberculin test (SIDT) and rapid assay kit. A total of 14 animals (2 males, 12 females) were found positive in rapid assay. In SIDT, four animals revealed a positive reaction in the neck region and seven animals in the tail base. Another male animal was found SIDT positive but negative in rapid assay; it died after 12 months. Nine rapid assay positive animals died asymptomatically in 1- to 11-month period revealing postmortem tuberculosis lesions that were confirmed by Ziehl-Neelsen staining and histopathology. No tuberculous lesion was evident in the animal found positive in SIDT alone. Results of the present study indicated that serological tests like rapid assay kit can serve as a reliable test for antemortem diagnosis of tuberculosis in dromedary camel.

  11. Force determination in lateral magnetic tweezers combined with TIRF microscopy.

    PubMed

    Madariaga-Marcos, J; Hormeño, S; Pastrana, C L; Fisher, G L M; Dillingham, M S; Moreno-Herrero, F

    2018-03-01

    Combining single-molecule techniques with fluorescence microscopy has attracted much interest because it allows the correlation of mechanical measurements with directly visualized DNA : protein interactions. In particular, its combination with total internal reflection fluorescence microscopy (TIRF) is advantageous because of the high signal-to-noise ratio this technique achieves. This, however, requires stretching long DNA molecules across the surface of a flow cell to maximize polymer exposure to the excitation light. In this work, we develop a module to laterally stretch DNA molecules at a constant force, which can be easily implemented in regular or combined magnetic tweezers (MT)-TIRF setups. The pulling module is further characterized in standard flow cells of different thicknesses and glass capillaries, using two types of micrometer size superparamagnetic beads, long DNA molecules, and a home-built device to rotate capillaries with mrad precision. The force range achieved by the magnetic pulling module was between 0.1 and 30 pN. A formalism for estimating forces in flow-stretched tethered beads is also proposed, and the results compared with those of lateral MT, demonstrating that lateral MT achieve higher forces with lower dispersion. Finally, we show the compatibility with TIRF microscopy and the parallelization of measurements by characterizing DNA binding by the centromere-binding protein ParB from Bacillus subtilis. Simultaneous MT pulling and fluorescence imaging demonstrate the non-specific binding of BsParB on DNA under conditions restrictive to condensation.

  12. Asymmetrical flow field-flow fractionation of white wine chromophoric colloidal matter.

    PubMed

    Coelho, Christian; Parot, Jérémie; Gonsior, Michael; Nikolantonaki, Maria; Schmitt-Kopplin, Philippe; Parlanti, Edith; Gougeon, Régis D

    2017-04-01

    Two analytical separation methods-size-exclusion chromatography and asymmetrical flow field-flow fractionation-were implemented to evaluate the integrity of the colloidal composition of Chardonnay white wine and the impact of pressing and fermentations on the final macromolecular composition. Wine chromophoric colloidal matter, representing UV-visible-absorbing wine macromolecules, was evaluated by optical and structural measurements combined with the description of elution profiles obtained by both separative techniques. The objective of this study was to apply these two types of fractionation on a typical Chardonnay white wine produced in Burgundy and to evaluate how each of them impacted the determination of the macromolecular chromophoric content of wine. UV-visible and fluorescence measurements of collected fractions were successfully applied. An additional proteomic study revealed that grape and microorganism proteins largely impacted the composition of chromophoric colloidal matter of Chardonnay wines. Asymmetrical flow field-flow fractionation appeared to be more reliable and less invasive with respect to the native chemical environment of chromophoric wine macromolecules, and hence is recommended as a tool to fractionate chromophoric colloidal matter in white wines. Graphical Abstract An innovative macromolecular separation method based on Asymmetrical Flow Field-Flow Fractionation was developed to better control colloidal dynamics across Chardonnay white winemaking.

  13. Simultaneous analysis for water- and fat-soluble vitamins by a novel single chromatography technique unifying supercritical fluid chromatography and liquid chromatography.

    PubMed

    Taguchi, Kaori; Fukusaki, Eiichiro; Bamba, Takeshi

    2014-10-03

    Chromatography techniques usually use a single state in the mobile phase, such as liquid, gas, or supercritical fluid. Chromatographers manage one of these techniques for their purpose but are sometimes required to use multiple methods, or even worse, multiple techniques when the target compounds have a wide range of chemical properties. To overcome this challenge, we developed a single method covering a diverse compound range by means of a "unified" chromatography which completely bridges supercritical fluid chromatography and liquid chromatography. In our method, the phase state was continuously changed in the following order; supercritical, subcritical and liquid. Moreover, the gradient of the mobile phase starting at almost 100% CO2 was replaced with 100% methanol at the end completely. As a result, this approach achieved further extension of the polarity range of the mobile phase in a single run, and successfully enabled the simultaneous analysis of fat- and water-soluble vitamins with a wide logP range of -2.11 to 10.12. Furthermore, the 17 vitamins were exceptionally separated in 4min. Our results indicated that the use of dense CO2 and the replacement of CO2 by methanol are practical approaches in unified chromatography covering diverse compounds. Additionally, this is a first report to apply the novel approach to unified chromatography, and can open another door for diverse compound analysis in a single chromatographic technique with single injection, single column and single system. Copyright © 2014. Published by Elsevier B.V.

  14. High temperature-ultra performance liquid chromatography-mass spectrometry for the metabonomic analysis of Zucker rat urine.

    PubMed

    Gika, Helen G; Theodoridis, Georgios; Extance, Jon; Edge, Anthony M; Wilson, Ian D

    2008-08-15

    The applicability and potential of using elevated temperatures and sub 2-microm porous particles in chromatography for metabonomics/metabolomics was investigated using, for the first time, solvent temperatures higher than the boiling point of water (up to 180 degrees C) and thermal gradients to reduce the use of organic solvents. Ultra performance liquid chromatography, combined with mass spectrometry, was investigated for the global metabolite profiling of the plasma and urine of normal and Zucker (fa/fa) obese rats (a well established disease animal model). "Isobaric" high temperature chromatography, where the temperature and flow rate follow a gradient program, was developed and evaluated against a conventional organic solvent gradient. LC-MS data were first examined by established chromatographic criteria in order to evaluate the chromatographic performance and next were treated by special peak picking algorithms to allow the application of multivariate statistics. These studies showed that, for urine (but not plasma), chromatography at elevated temperatures provided better results than conventional reversed-phase LC with higher peak capacity and better peak asymmetry. From a systems biology point of view, better group clustering and separation was obtained with a larger number of variables of high importance when using high temperature-ultra performance liquid chromatography (HT-UPLC) compared to conventional solvent gradients.

  15. Particle size effects on protein and virus-like particle adsorption on perfusion chromatography media.

    PubMed

    Wu, Yige; Abraham, Dicky; Carta, Giorgio

    2015-01-02

    The resin structure, chromatographic behavior, and adsorption kinetics of proteins and virus-like-particles (VLPs) are studied for POROS HS 20 and POROS HS 50 (23 and 52 μm mean diameter, respectively) to determine the effects of particle size on perfusion chromatography and to determine the predictive ability of available models. Transmission electron microscopy (TEM) and inverse size-exclusion chromatography (iSEC) show similar structures for the two resins, both containing 200-1000 nm pores that transect a network of much smaller pores. For non-binding conditions, trends of the height equivalent to a theoretical plate (HETP) as a function of reduced velocity are consistent with perfusion. The estimated intraparticle flow fractions for these conditions are 0.0018 and 0.00063 for POROS HS 20 and HS 50, respectively. For strong binding conditions, confocal laser scanning microscopy (CLSM) shows asymmetrical intraparticle concentrations profiles and enhanced rates of IgG adsorption on POROS HS 20 at 1000 cm/h. The corresponding effective diffusivity under flow is 2-3 times larger than for non-flow conditions and much larger than observed for POROS HS 50, consistent with available models. For VLPs, however, adsorption is confined to a thin layer near the particle surface for both resins, suggesting that the bound VLPs block the pores. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. A rapid lateral-flow immunoassay for phytosanitary detection of Erwinia amylovora and on-site fire blight diagnosis.

    PubMed

    Braun-Kiewnick, A; Altenbach, D; Oberhänsli, T; Bitterlin, W; Duffy, B

    2011-10-01

    Fire blight is an invasive disease caused by Erwinia amylovora that threatens pome fruit production globally. Effective implementation of phytosanitary control measures depends upon rapid, reliable pathogen detection and disease diagnosis. We developed a lateral-flow immunoassay specific for E. amylovora with a detection limit of log 5.7 CFU/ml, typical of pathogen concentrations in symptomatic plant material. The simple assay had comparable sensitivity to standard culture plating, serum agglutination and nested PCR when validated for application in a phytosanitary laboratory as a confirmatory test of cultured isolates and for first-line diagnosis of phytosanitary samples that represent the full range of commercial, ornamental and forestry host species. On-site validation in ring-trials with local plant inspectors demonstrated robust and reliable detection (compared to subsequent plating and PCR analysis). The simplicity, inspector acceptance and facilitation of expedited diagnosis (from 2 days for laboratory submitted samples to 15 min with the immunoassay), offers a valuable tool for improved phytosanitary control of fire blight. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Multiple Cross Displacement Amplification Combined with Gold Nanoparticle-Based Lateral Flow Biosensor for Detection of Vibrio parahaemolyticus

    PubMed Central

    Wang, Yi; Li, Hui; Li, Dongxun; Li, Kewei; Wang, Yan; Xu, Jianguo; Ye, Changyun

    2016-01-01

    Vibrio parahaemolyticus (V. parahaemolyticus) is a marine seafood-borne pathogen causing severe illnesses in humans and aquatic animals. In the present study, multiple cross displacement amplification was combined with a lateral flow biosensor (MCDA-LFB) to detect the toxR gene of V. parahaemolyticus in DNA extracts from pure cultures and spiked oyster homogenates. Amplification was carried out at a constant temperature (62°C) for only 30 min, and amplification products were directly applied to the biosensor. The entire process, including oyster homogenate processing (30 min), isothermal amplification (30 min) and results indicating (∼2 min), could be completed within 65 min. Amplification product was detectable from as little as 10 fg of pure V. parahaemolyticus DNA and from approximately 4.2 × 102 CFU in 1 mL of oyster homogenate. No cross-reaction with other Vibrio species and with non-Vibrio species was observed. Therefore, the MCDA-LFB method established in the current report is suitable for the rapid screening of V. parahaemolyticus in clinical, food, and environmental samples. PMID:28066368

  18. Simultaneous determination of Cr(III) and Cr(VI) in tannery wastewater using low pressure ion chromatography combined with flow injection spectrophotometry

    NASA Astrophysics Data System (ADS)

    Chen, Shujuan; Zhang, Xinshen; Yu, Lingyun; Wang, Li; Li, Hui

    2012-03-01

    Trivalent and hexavalent chromium have been successfully separated and determined using low pressure ion chromatography combined with flow injection spectrophotometric analysis (LPIC-FIA). A column packed with crosslinking starch microspheres was used for on-line separation of Cr(III) from Cr(VI) in a flow-injection system because of its absorptive effect on Cr(III). To determine the concentration of Cr(III) and Cr(VI) in samples, we used 3.0 mmol/L nitric acid to elute adsorbed Cr(III) from the column and then used ceric sulfate-sulfuric acid as oxidant to convert all Cr(III) into Cr(VI). Then, Cr(VI) directly came from the samples and Cr(VI) came from Cr(III) successively formed a amaranthine complex with diphenycarbazide and the complex shows a maximum absorption at 530 nm. Analytical parameters including the concentration of eluent and oxidant solution, oxidizing temperature, length of oxidizing reaction coil, reaction coil and injection coil, interfering effects, etc., were optimized. The limit of detection was 1.25 μg/L for Cr(VI) and 3.76 μg/L for Cr(III). The linear relationship between absorption with the concentration of Cr(VI) and Cr(III) was 0.001-1.000 mg/L and 0.030-1.000 mg/L with correlation coefficients of 0.9995 and 0.9994, respectively. The relative standard deviation of Cr(VI) and Cr(III) was 1.21% and 1.66%, respectively (n = 10). Major cations and anions did not show any interference. We validated this method through certified reference materials and through measuring the recovery in tannery wastewater.

  19. Characterization of phenolic amides from cortex lycii by ultra high-performance liquid chromatography coupled with LTQ-Orbitrap mass spectrometry

    USDA-ARS?s Scientific Manuscript database

    High performance liquid chromatography (UPLC) and flow injection electrospray ionization with ion trap mass spectrometry (FIMS) fingerprints combined with the principal component analysis (PCA) were examined for their potential in differentiating commercial organic and conventional sage samples. The...

  20. Influence of slip-surface geometry on earth-flow deformation, Montaguto earth flow, southern Italy

    USGS Publications Warehouse

    Guerriero, L.; Coe, Jeffrey A.; Revellio, P.; Grelle, G.; Pinto, F.; Guadagno, F.

    2016-01-01

    We investigated relations between slip-surface geometry and deformational structures and hydrologic features at the Montaguto earth flow in southern Italy between 1954 and 2010. We used 25 boreholes, 15 static cone-penetration tests, and 22 shallow-seismic profiles to define the geometry of basal- and lateral-slip surfaces; and 9 multitemporal maps to quantify the spatial and temporal distribution of normal faults, thrust faults, back-tilted surfaces, strike-slip faults, flank ridges, folds, ponds, and springs. We infer that the slip surface is a repeating series of steeply sloping surfaces (risers) and gently sloping surfaces (treads). Stretching of earth-flow material created normal faults at risers, and shortening of earth-flow material created thrust faults, back-tilted surfaces, and ponds at treads. Individual pairs of risers and treads formed quasi-discrete kinematic zones within the earth flow that operated in unison to transmit pulses of sediment along the length of the flow. The locations of strike-slip faults, flank ridges, and folds were not controlled by basal-slip surface topography but were instead dependent on earth-flow volume and lateral changes in the direction of the earth-flow travel path. The earth-flow travel path was strongly influenced by inactive earth-flow deposits and pre-earth-flow drainages whose positions were determined by tectonic structures. The implications of our results that may be applicable to other earth flows are that structures with strikes normal to the direction of earth-flow motion (e.g., normal faults and thrust faults) can be used as a guide to the geometry of basal-slip surfaces, but that depths to the slip surface (i.e., the thickness of an earth flow) will vary as sediment pulses are transmitted through a flow.

  1. Two-dimensional chromatography of complex polymers, 8. Separation of fatty alcohol ethoxylates simultaneously by end group and chain length.

    PubMed

    Raust, Jacques-Antoine; Bruell, Adele; Sinha, Pritish; Hiller, Wolf; Pasch, Harald

    2010-09-01

    A comprehensive two-dimensional liquid chromatography system was developed to precisely describe the molecular heterogeneity of fatty alcohol ethoxylates. The end-group functionality was analyzed by gradient HPLC while ethylene oxide oligomer distributions were characterized by liquid adsorption chromatography. A baseline separation of all functionality fractions irrespective of the ethylene oxide oligomer chain length was achieved on nonpolar X-Terra(®) C(18) with a methanol-water gradient, whereas an isocratic flow of isopropanol-water on a polar Chromolith(®) Si column gave a separation according to the oligomer chain length without interference of the end-group distribution. The combination of these two methods to conduct online two-dimensional liquid chromatography experiments resulted in a comprehensive two-dimensional picture on the molecular heterogeneity of the sample.

  2. Twin tubular pinch effect in curving confined flows

    PubMed Central

    Clime, Liviu; Morton, Keith J.; Hoa, Xuyen D.; Veres, Teodor

    2015-01-01

    Colloidal suspensions of buoyancy neutral particles flowing in circular pipes focus into narrow distributions near the wall due to lateral migration effects associated with fluid inertia. In curving flows, these distributions are altered by Dean currents and the interplay between Reynolds and Dean numbers is used to predict equilibrium positions. Here, we propose a new description of inertial lateral migration in curving flows that expands current understanding of both focusing dynamics and equilibrium distributions. We find that at low Reynolds numbers, the ratio δ between lateral inertial migration and Dean forces scales simply with the particle radius, coil curvature and pipe radius as . A critical value δc = 0.148 of this parameter is identified along with two related inertial focusing mechanisms. In the regime below δc, coined subcritical, Dean forces generate permanently circulating, twinned annuli, each with intricate equilibrium particle distributions including eyes and trailing arms. At δ > δc (supercritical regime) inertial lateral migration forces are dominant and particles focus to a single stable equilibrium position. PMID:25927878

  3. Design and performance of a new continuous-flow sample-introduction system for flame infrared-emission spectrometry: Applications in process analysis, flow injection analysis, and ion-exchange high-performance liquid chromatography.

    PubMed

    Lam, C K; Zhang, Y; Busch, M A; Busch, K W

    1993-06-01

    A new sample introduction system for the analysis of continuously flowing liquid streams by flame infrared-emission (FIRE) spectrometry has been developed. The system uses a specially designed purge cell to strip dissolved CO(2) from solution into a hydrogen gas stream that serves as the fuel for a hydrogen/air flame. Vibrationally excited CO(2) molecules present in the flame are monitored with a simple infrared filter (4.4 mum) photometer. The new system can be used to introduce analytes as a continuous liquid stream (process analysis mode) or on a discrete basis by sample injection (flow injection analysis mode). The key to the success of the method is the new purge-cell design. The small internal volume of the cell minimizes problems associated with purge-cell clean-out and produces sharp, reproducible signals. Spent analytical solution is continuously drained from the cell, making cell disconnection and cleaning between samples unnecessary. Under the conditions employed in this study, samples could be analyzed at a maximum rate of approximately 60/h. The new sample introduction system was successfully tested in both a process analysis- and a flow injection analysis mode for the determination of total inorganic carbon in Waco tap water. For the first time, flame infrared-emission spectrometry was successfully extended to non-volatile organic compounds by using chemical pretreatment with peroxydisulfate in the presence of silver ion to convert the analytes into dissolved carbon dioxide, prior to purging and detection by the FIRE radiometer. A test of the peroxydisulfate/Ag(+) reaction using six organic acids and five sugars indicated that all 11 compounds were oxidized to nearly the same extent. Finally, the new sample introduction system was used in conjunction with a simple filter FIRE radiometer as a detection system in ion-exchange high-performance liquid chromatography. Ion-exchange chromatograms are shown for two aqueous mixtures, one containing six organic

  4. Rapid Detection of Escherichia coli O157 and Shiga Toxins by Lateral Flow Immunoassays

    PubMed Central

    Wang, Jinliang; Katani, Robab; Li, Lingling; Hegde, Narasimha; Roberts, Elisabeth L.; Kapur, Vivek; DebRoy, Chitrita

    2016-01-01

    Shiga toxin-producing Escherichia coli O157:H7 (STEC) cause food-borne illness that may be fatal. STEC strains enumerate two types of potent Shiga toxins (Stx1 and Stx2) that are responsible for causing diseases. It is important to detect the E. coli O157 and Shiga toxins in food to prevent outbreak of diseases. We describe the development of two multi-analyte antibody-based lateral flow immunoassays (LFIA); one for the detection of Stx1 and Stx2 and one for the detection of E. coli O157 that may be used simultaneously to detect pathogenic E. coli O157:H7. The LFIA strips were developed by conjugating nano colloidal gold particles with monoclonal antibodies against Stx1 and Stx2 and anti-lipid A antibodies to capture Shiga toxins and O157 antigen, respectively. Our results indicate that the LFIA for Stx is highly specific and detected Stx1 and Stx2 within three hours of induction of STEC with ciprofloxacin at 37 °C. The limit of detection for E. coli O157 LFIA was found to be 105 CFU/mL in ground beef spiked with the pathogen. The LFIAs are rapid, accurate and easy to use and do not require sophisticated equipment or trained personnel. Following the assay, colored bands on the membrane develop for end-point detection. The LFIAs may be used for screening STEC in food and the environment. PMID:27023604

  5. Mechanical characterisation of agarose-based chromatography resins for biopharmaceutical manufacture.

    PubMed

    Nweke, Mauryn C; McCartney, R Graham; Bracewell, Daniel G

    2017-12-29

    Mechanical characterisation of agarose-based resins is an important factor in ensuring robust chromatographic performance in the manufacture of biopharmaceuticals. Pressure-flow profiles are most commonly used to characterise these properties. There are a number of drawbacks with this method, including the potential need for several re-packs to achieve the desired packing quality, the impact of wall effects on experimental set up and the quantities of chromatography media and buffers required. To address these issues, we have developed a dynamic mechanical analysis (DMA) technique that characterises the mechanical properties of resins based on the viscoelasticity of a 1ml sample of slurry. This technique was conducted on seven resins with varying degrees of mechanical robustness and the results were compared to pressure-flow test results on the same resins. Results show a strong correlation between the two techniques. The most mechanically robust resin (Capto Q) had a critical velocity 3.3 times higher than the weakest (Sepharose CL-4B), whilst the DMA technique showed Capto Q to have a slurry deformation rate 8.3 times lower than Sepharose CL-4B. To ascertain whether polymer structure is indicative of mechanical strength, scanning electron microscopy images were also used to study the structural properties of each resin. Results indicate that DMA can be used as a small volume, complementary technique for the mechanical characterisation of chromatography media. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

  6. Behavior of lateral-deformation coefficients during elastoplastic deformation of metals

    NASA Astrophysics Data System (ADS)

    Zimin, B. A.; Smirnov, I. V.; Sudenkov, Yu. V.

    2017-06-01

    The results of investigations into variation of the coefficients of lateral deformation (the Poisson ratio) during single-axis tension of samples of steel 12Kh18N10T and St3, titanium VT1, the aluminum alloy D16AM, copper M1, and a magnesium alloy are considered. The technique developed on the basis of the optoacoustic effect and simultaneous measurements of the longitudinal and surface speeds of sound in metallic samples during the tension makes it possible to measure the rates at various stages of the deformation process. The data obtained make it possible to construct the dependences of variation of the lateral-deformation coefficients at all stages of the plastic flow. The correlation of these variations both with known processes of structural reconstructions at various stages of plastic flow and with the process of localization of plastic-shear bands in the aluminum alloy is noted.

  7. Evaluation of a Newly Developed Lateral Flow Immunoassay for the Diagnosis of Cryptococcosis

    PubMed Central

    Lindsley, Mark D.; Mekha, Nanthawan; Baggett, Henry C.; Surinthong, Yupha; Autthateinchai, Rinrapas; Sawatwong, Pongpun; Harris, Julie R.; Park, Benjamin J.; Chiller, Tom; Poonwan, Natteewan

    2011-01-01

    Background. Cryptococcosis is a common opportunistic infection of human immunodeficiency virus (HIV)–infected individuals mostly occurring in resource-limited countries. This study compares the performance of a recently developed lateral flow immunoassay (LFA) to blood culture and enzyme immunoassay (EIA) for the diagnosis of cryptococcosis. Methods. Archived sera from 704 HIV-infected patients hospitalized for acute respiratory illness in Thailand were tested for cryptococcal antigenemia using EIA. All EIA-positive and a subset of EIA-negative sera were tested by LFA, with results recorded after 5 and 15 minutes incubation. Urine from patients with LFA- and EIA-positive sera was tested by LFA. Antigen results from patients with positive cryptococcal blood cultures were compared. Results. Of 704 sera, 92 (13%) were positive by EIA; among the 91 EIA-positive sera tested by LFA, 82 (90%) and 87 (96%) were LFA positive when read after 5 and 15 minutes, respectively. Kappa agreement of EIA and LFA for sera was 0.923 after 5 minutes and 0.959 after 15 minutes, respectively. Two of 373 EIA-negative sera were LFA positive at both time points. Of 74 urine specimens from EIA-positive patients, 52 (70.3%) were LFA positive. EIA was positive in 16 of 17 sera from blood culture–positive patients (94% sensitivity), and all sera were positive by LFA (100% sensitivity). Conclusions. A high level of agreement was shown between LFA and EIA testing of serum. The LFA is a rapid, easy-to-perform assay that does not require refrigeration, demonstrating its potential usefulness as a point-of-care assay for diagnosis of cryptococcosis in resource-limited countries. PMID:21810743

  8. [Determination and analysis of toluene diisocyanate metabolites in mice using gas chromatography-mass spectrometry].

    PubMed

    Ji, Yu-Bin; Ji, Chen-Feng; Zou, Xiang; Liu, Hui-Xin

    2007-09-01

    In the present research we used gas chromatography-mass spectrometry (G C-MS) to determine metabolites of toluene diisocyanate (TDI) in mice and deduce the pathway for toluene diisocyanate metabolism in the organism. Conditions for TDI chromatography: Supelco PTETM-5 chromatographic column (30 mm x 0.25 mm x 0.25 microm); initial column temperature: 90 degrees C, which was maintained for 30 min, then the temperature was increased at a rate of 40 degrees C x min(-1) to 280 degrees C, and maintained for 5.25 min; temperature for the vaporizing chamber: 250 degrees C; carrier gas: helium flowing at 1.0 microL x min(-1). Conditions for chromatography of TDI metabolites in the organism: 94% methyl, 4% ethenyl-bonded-phase fused-silica capillary column (30 + 2 m x 0.25 + 0.02 mm); initial column temperature: 30 degrees C, which was maintained for 5 min, after and then was increased at a rate of 80 degrees C x min(-1) to 280 degrees C, and maintained for 5 min; temperature for the vaporizing chamber: 250 degrees C; carrier gas: helium flowing at 1.0 microL x min(-1). Conditions for mass spectrometry: EI for ionization; 70 eV for ionization energy; 280 degrees C for connecting tube temperature; 35-350 micro for range of scanning; and 1.0 microL for sample size. The results showed that 2 ,4-toluene diisocyanate was metabolized into 2,4-diaminotoluene. Under the conditions selected for GC-MS, TDI metabolites in the organism can be isolated and identified.

  9. Lateral migration of a microdroplet under optical forces in a uniform flow

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cho, Hyunjun; Chang, Cheong Bong; Jung, Jin Ho

    2014-12-15

    The behavior of a microdroplet in a uniform flow and subjected to a vertical optical force applied by a loosely focused Gaussian laser beam was studied numerically. The lattice Boltzmann method was applied to obtain the two-phase flow field, and the dynamic ray tracing method was adopted to calculate the optical force. The optical forces acting on the spherical droplets agreed well with the analytical values. The numerically predicted droplet migration distances agreed well with the experimentally obtained values. Simulations of the various flow and optical parameters showed that the droplet migration distance nondimensionalized by the droplet radius is proportionalmore » to the S number (z{sub d}/r{sub p} = 0.377S), which is the ratio of the optical force to the viscous drag. The effect of the surface tension was also examined. These results indicated that the surface tension influenced the droplet migration distance to a lesser degree than the flow and optical parameters. The results of the present work hold for the refractive indices of the mean fluid and the droplet being 1.33 and 1.59, respectively.« less

  10. Liquid Chromatography in 1982.

    ERIC Educational Resources Information Center

    Freeman, David H.

    1982-01-01

    Reviews trends in liquid chromatography including apparatus, factors affecting efficient separation of a mixture (peak sharpness and speed), simplified problem-solving, adsorption, bonded phase chromatography, ion selectivity, and size exclusion. The current trend is to control chemical selectivity by the liquid phase. (Author/JN)

  11. Freeze chromatography method and apparatus

    DOEpatents

    Scott, C.D.

    1987-04-16

    A freeze chromatography method and apparatus are provided which enable separation of the solutes contained in a sample. The apparatus includes an annular column construction comprising cylindrical inner and outer surfaces defining an annular passage therebetween. One of the surfaces is heated and the other cooled while passing an eluent through the annular passageway so that the eluent in contact with the cooled surface freezes and forms a frozen eluent layer thereon. A mixture of solutes dissolved in eluent is passed through the annular passageway in contact with the frozen layer so that the sample solutes in the mixture will tend to migrate either toward or away the frozen layer. The rate at which the mixture flows through the annular passageway is controlled so that the distribution of the sample solutes approaches that at equilibrium and thus a separation between the sample solutes occurs. 3 figs.

  12. Sample stream distortion modeled in continuous-flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Rhodes, P. H.

    1979-01-01

    Buoyancy-induced disturbances in an electrophoresis-type chamber were investigated. Five tracer streams (latex) were used to visualize the flows while a nine-thermistor array sensed the temperature field. The internal heating to the chamber was provided by a 400 Hz electrical field. Cooling to the chamber was provided on the front and back faces and, in addition, on both chamber side walls. Disturbances to the symmetric base flow in the chamber occurred in the broad plane of the chamber and resulted from the formation of lateral and axial temperature gradients. The effect of these gradients was to retard or increase local flow velocities at different positions in the chamber cross section, which resulted in lateral secondary flows being induced in the broad plane of the chamber. As the adverse temperature gradients increased in magnitude, the critical Rayleigh number was approached and reverse (separated) flow became apparent, which, subsequently, led to the onset of time variant secondary flows.

  13. Enantioselective comprehensive two-dimensional gas chromatography of lavender essential oil.

    PubMed

    Krupčík, Ján; Gorovenko, Roman; Špánik, Ivan; Armstrong, Daniel W; Sandra, Pat

    2016-12-01

    The enantiomeric composition of several chiral markers in lavender essential oil was studied by flow modulated comprehensive two-dimensional gas chromatography operated in the reverse flow mode and hyphenated to flame ionization and quadrupole mass spectrometric detection. Two capillary column series were used in this study, 2,3-di-O-ethyl-6-O-tert-butyldimethylsilyl-β-cyclodextrin or 2,3,6-tri-O-methyl-β-cyclodextrin, as the chiral column in the first dimension and α polyethylene glycol column in the second dimension. Combining the chromatographic data obtained on these column series, the enantiomeric and excess ratios for α-pinene, β-pinene, camphor, lavandulol, borneol, and terpinen-4-ol were determined. This maybe a possible route to assess the authenticity of lavender essential oil. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Development of a lateral flow colloidal gold immunoassay strip for the rapid detection of enrofloxacin residues.

    PubMed

    Zhao, Yinli; Zhang, Gaiping; Liu, Qingtang; Teng, Man; Yang, Jifei; Wang, Jianhua

    2008-12-24

    A rapid immunochromatographic lateral flow test strip of competitive format has been developed using a gold-conjugated monoclonal antibody for the specific determination of enrofloxacin (ENR) residues in chicken muscles. For this purpose, a specific monoclonal antibody (mAb) for ENR was generated and characterized. The mAb showed negligible cross-reactivity with other related compounds. Using ENR standards prepared in chicken muscle extracts from 0 to 24.3 ng/mL (microg/kg), the method indicated that the detection limit of the test strip, as measured in a strip scanner, was as low as 0.138 microg/kg of ENR and the half-maximal inhibition concentration (IC(50)) was 0.935 microg/kg. For samples spiked at 10, 20, and 30 microg/kg, the recovery was between 85.3 and 96.1% and the coefficient of variation [CV (%)] was between 4.5 and 7.91%. Parallel analysis of muscle samples from chickens fed ENR showed good comparable results obtained from the test strip and LC-MS. Each test requires 5-10 min. The data indicate that the method has high sensitivity, specificity, and the advantages of simplicity and speed of performance. Therefore, the test strip provides a useful screening method for quantitative, semiquantitative, or qualitative detection of ENR residues in chicken muscles.

  15. Recombinase polymerase amplification combined with lateral flow dipstick for equipment-free detection of Salmonella in shellfish.

    PubMed

    Gao, Weifang; Huang, Hailong; Zhu, Peng; Yan, Xiaojun; Fan, Jianzhong; Jiang, Jinpo; Xu, Jilin

    2018-05-01

    Salmonella is a major pathogen that causes acute foodborne outbreaks worldwide. Seafood, particularly shellfish, is a proven source of Salmonella spp. infection because many people prefer to eat it raw or lightly cooked. However, traditional identification methods are too time-consuming and complex to detect contamination of bacteria in the food chain in a timely manner, and few studies have aimed to identify Salmonella in shellfish early in the supply chain. We herein developed a method for rapid detection of Salmonella in shellfish based on the method of recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD), which targets the invasion gene A (invA). The RPA-LFD was able to function at 30-45 °C, and at the temperature of 40 °C, it only took 8 min of amplification to reach the test threshold of amplicons. The established method had both a good specificity and a sensitivity of 100 fg DNA per reaction (20 µL). Regarding practical performance, RPA-LFD performed better than real-time PCR. Another advantage of RPA-LFD is that it was capable of being performed without expensive equipments. Thus, RPA-LFD has potential for further development as a detection kit for Salmonella in shellfish and other foods under field conditions.

  16. 3D-printed and CNC milled flow-cells for chemiluminescence detection.

    PubMed

    Spilstead, Kara B; Learey, Jessica J; Doeven, Egan H; Barbante, Gregory J; Mohr, Stephan; Barnett, Neil W; Terry, Jessica M; Hall, Robynne M; Francis, Paul S

    2014-08-01

    Herein we explore modern fabrication techniques for the development of chemiluminescence detection flow-cells with features not attainable using the traditional coiled tubing approach. This includes the first 3D-printed chemiluminescence flow-cells, and a milled flow-cell designed to split the analyte stream into two separate detection zones within the same polymer chip. The flow-cells are compared to conventional detection systems using flow injection analysis (FIA) and high performance liquid chromatography (HPLC), with the fast chemiluminescence reactions of an acidic potassium permanganate reagent with morphine and a series of adrenergic phenolic amines. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Promising Nucleic Acid Lateral Flow Assay Plus PCR for Shiga Toxin-Producing Escherichia coli.

    PubMed

    Terao, Yoshitaka; Takeshita, Kana; Nishiyama, Yasutaka; Morishita, Naoki; Matsumoto, Takashi; Morimatsu, Fumiki

    2015-08-01

    Shiga toxin (Stx)-producing Escherichia coli (STEC) is a frequent cause of foodborne infections, and methods for rapid and reliable detection of STEC are needed. A nucleic acid lateral flow assay (NALFA) plus PCR was evaluated for detecting STEC after enrichment. When cell suspensions of 45 STEC strains, 14 non-STEC strains, and 13 non-E. coli strains were tested with the NALFA plus PCR, all of the STEC strains yielded positive results, and all of the non-STEC and non-E. coli strains yielded negative results. The lower detection limit for the STEC strains ranged from 0.1 to 1 pg of genomic DNA (about 20 to 200 CFU) per test, and the NALFA plus PCR was able to detect Stx1- and Stx2-producing E. coli strains with similar sensitivities. The ability of the NALFA plus PCR to detect STEC in enrichment cultures of radish sprouts, tomato, raw ground beef, and beef liver inoculated with 10-fold serially diluted STEC cultures was comparable to that of a real-time PCR assay (at a level of 100 to 100,000 CFU/ml in enrichment culture). The bacterial inoculation test in raw ground beef revealed that the lower detection limit of the NALFA plus PCR was also comparable to that obtained with a real-time PCR assay that followed the U.S. Department of Agriculture guidelines. Although further evaluation is required, these results suggest that the NALFA plus PCR is a specific and sensitive method for detecting STEC in a food manufacturing plant.

  18. Multimodal charge-induction chromatography for antibody purification.

    PubMed

    Tong, Hong-Fei; Lin, Dong-Qiang; Chu, Wen-Ning; Zhang, Qi-Lei; Gao, Dong; Wang, Rong-Zhu; Yao, Shan-Jing

    2016-01-15

    Hydrophobic charge-induction chromatography (HCIC) has advantages of high capacity, salt-tolerance and convenient pH-controlled elution. However, the binding specificity might be improved with multimodal molecular interactions. New ligand W-ABI that combining tryptophan and 5-amino-benzimidazole was designed with the concept of mutimodal charge-induction chromatography (MCIC). The indole and benzimidazole groups of the ligand could provide orientated mutimodal binding to target IgG under neutral pH, while the imidazole groups could induce the electrostatic repulsion forces for efficient elution under acidic pH. W-ABI ligand was coupled successfully onto agarose gel, and IgG adsorption behaviors were investigated. High affinity to IgG was found with the saturated adsorption capacity of 70.4 mg/ml at pH 7, and the flow rate of mobile phase showed little impact on the dynamic binding capacity. In addition, efficient elution could be achieved at mild acidic pH with high recovery. Two separation cases (IgG separation from albumin containing feedstock and monoclonal antibody purification from cell culture supernatant) were verified with high purity and recovery. In general, MCIC with the specially-designed ligand is an expanding of HCIC with improved adsorption selectivity, which would be a potential alternative to Protein A-based capture for the cost-effective purification of antibodies. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Purification of bacteriophage M13 by anion exchange chromatography.

    PubMed

    Monjezi, Razieh; Tey, Beng Ti; Sieo, Chin Chin; Tan, Wen Siang

    2010-07-01

    M13 is a non-lytic filamentous bacteriophage (phage). It has been used widely in phage display technology for displaying foreign peptides, and also for studying macromolecule structures and interactions. Traditionally, this phage has been purified by cesium chloride (CsCl) density gradient ultracentrifugation which is highly laborious and time consuming. In the present study, a simple, rapid and efficient method for the purification of M13 based on anion exchange chromatography was established. A pre-packed SepFast Super Q column connected to a fast protein liquid chromatography (FPLC) system was employed to capture released phages in clarified Escherichia coli fermented broth. An average yield of 74% was obtained from a packed bed mode elution using citrate buffer (pH 4), containing 1.5 M NaCl at 1 ml/min flow rate. The purification process was shortened substantially to less than 2 h from 18 h in the conventional ultracentrifugation method. SDS-PAGE revealed that the purity of particles was comparable to that of CsCl gradient density ultracentrifugation method. Plaque forming assay showed that the purified phages were still infectious. Copyright 2010 Elsevier B.V. All rights reserved.

  20. A high pressure liquid chromatography method for separation of prolactin forms.

    PubMed

    Bell, Damon A; Hoad, Kirsten; Leong, Lillian; Bakar, Juwaini Abu; Sheehan, Paul; Vasikaran, Samuel D

    2012-05-01

    Prolactin has multiple forms and macroprolactin, which is thought not to be bioavailable, can cause a raised serum prolactin concentration. Gel filtration chromatography (GFC) is currently the gold standard method for separating macroprolactin, but is labour-intensive. Polyethylene glycol (PEG) precipitation is suitable for routine use but may not always be accurate. We developed a high pressure liquid chromatography (HPLC) assay for macroprolactin measurement. Chromatography was carried out using an Agilent Zorbax GF-250 (9.4 × 250 mm, 4 μm) size exclusion column and 50 mmol/L Tris buffer with 0.15 mmol/L NaCl at pH 7.2 as mobile phase, with a flow rate of 1 mL/min. Serum or plasma was diluted 1:1 with mobile phase and filtered and 100 μL injected. Fractions of 155 μL were collected for prolactin measurement and elution profile plotted. The area under the curve of each prolactin peak was calculated to quantify each prolactin form, and compared with GFC. Clear separation of monomeric-, big- and macroprolactin forms was achieved. Quantification was comparable to GFC and precision was acceptable. Total time from injection to collection of the final fraction was 16 min. We have developed an HPLC method for quantification of macroprolactin, which is rapid and easy to perform and therefore can be used for routine measurement.

  1. Impact of lateral flow on the transition from connected to disconnected stream-aquifer systems

    NASA Astrophysics Data System (ADS)

    Xian, Yang; Jin, Menggui; Liu, Yanfeng; Si, Aonan

    2017-05-01

    Understanding the mechanisms by which stream water infiltrates through streambeds to recharge groundwater systems is essential to sustainable management of scarce water resources in arid and semi-arid areas. An inverted water table (IWT) can develop under a stream in response to the desaturation between the stream and underlying aquifer as the system changes from a connected to disconnected status. However, previous studies have suggested that the IWT can only occur at the bottom of a low permeability streambed in which only the vertical flow between the stream and groundwater during disconnection was assumed. In the present study, numerical simulations revealed that the lateral flow induced by capillarity or heterogeneity also plays an essential role on interactions between streams and aquifers. Three pathways were identified for the transition from connection to disconnection in homogenous systems; notably, the lowest point of an IWT can develop not only at the bottom of the streambed but also within the streambed or the aquifer in response to the initial desaturation at, above, or below the interface between the streambed and aquifer (IBSA), respectively. A sensitivity analysis indicated that in wide streams, the lowest point of an IWT only occurs at the bottom of the streambed; however, for a stream half width of 1 m above a 6 m thick sandy loam streambed, the lowest point occurs in the streambed as stream depth is less than 0.5 m. This critical stream depth increases with streambed thickness and decreases with stream width. Thus, in narrow streams the lowest point can also develop in a thick streambed under a shallow stream. In narrow streams, the lowest point also forms in the aquifer if the ratio of the hydraulic conductivity of the streambed to that of the aquifer is greater than the ratio of the streambed thickness to the sum of the stream depth and the streambed thickness; correspondingly, the streambed is thin but relatively permeable and the stream is

  2. Rapid diagnosis of cryptococcal meningitis by use of lateral flow assay on cerebrospinal fluid samples: influence of the high-dose "hook" effect.

    PubMed

    Lourens, Adré; Jarvis, Joseph N; Meintjes, Graeme; Samuel, Catherine M

    2014-12-01

    Cryptococcal meningitis is the most frequent cause of meningitis and a major cause of mortality in HIV-infected adults in Africa. This study evaluated the performance of the lateral flow assay (LFA) on cerebrospinal fluid (CSF) samples for the diagnosis of cryptococcal meningitis against that of existing diagnostic tests. LFA performed on 465 undiluted CSF samples had a sensitivity of 91%. When the LFA was paired with Gram staining, a sensitivity of 100% was achieved after implementation of a dilution step for samples with negative LFA results and the presence of yeasts on microscopy. Microscopy is essential for preventing the reporting of false-negative results due to the high-dose "hook" effect. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  3. Effect of High-Frequency Oscillations on Cough Peak Flows Generated by Mechanical In-Exsufflation in Medically Stable Subjects With Amyotrophic Lateral Sclerosis.

    PubMed

    Sancho, Jesús; Bures, Enric; de La Asunción, Saray; Servera, Emilio

    2016-08-01

    Mechanically assisted coughing with mechanical in-exsufflation (MI-E) is recommended for noninvasive management of respiratory secretions in amyotrophic lateral sclerosis (ALS). To improve the effectiveness of the technique, a new device combining MI-E with high-frequency oscillations (HFO) has been developed. This work aimed to assess the effect of HFO on the cough peak flow generated by MI-E in medically stable subjects with ALS. This was a prospective study that included subjects with ALS in a medically stable condition. Cough peak flow generated by MI-E was measured in 4 situations: without HFO, with HFO during insufflation, with HFO during exsufflation, and with HFO in both cycles. The parameters used were: insufflation pressure of +40 cm H2O, exsufflation pressure of -40 cm H2O, insufflation time 2 s, exsufflation time 3 s, amplitude of oscillations 10 cm H2O, and frequency of oscillations 15 Hz. Forty-seven subjects with ALS were included: 66% males, 68.2 ± 9.2 y, 40% with bulbar onset, FVC = 1.7 ± 1.1 L, percent-of-predicted FVC = 54.4 ± 26.6%, cough peak flow = 3.8 ± 2.2 L/s, PImax = -39.4 ± 26.4 cm H2O, revised ALS scale = 28.5 ± 9.3, Norris bulbar subscore = 26.1 ± 10.4. No statistical differences were found in cough peak flow generated by MI-E in the 4 situations (without HFO = 4.0 ± 1.2 L/s, with insufflation HFO = 3.9 ± 1.2 L/s, with exsufflation HFO = 4.1 ± 1.2 L/s, with in-exsufflation HFO = 3.9 ± 1.1 L/s). The addition of HFO to mechanically assisted coughing with MI-E does not have an effect on the cough peak flow of medically stable subjects with ALS. Copyright © 2016 by Daedalus Enterprises.

  4. Increasing Sensitivity In Continuous-Flow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Sharnez, Rizwan; Sammons, David W.

    1994-01-01

    Sensitivity of continuous-flow electrophoresis (CFE) chamber increased by introducing lateral gradients in concentration of buffer solution and thickness of chamber. Such gradients, with resulting enhanced separation, achieved in CFE chamber with wedge-shaped cross section and collateral flow. Enables improved separations of homogeneous components of mixtures of variety of biologically important substances.

  5. Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

    PubMed Central

    Murphy, Patrick J. M.; Stone, Orrin J.; Anderson, Michelle E.

    2011-01-01

    In contrast to other chromatographic methods for purifying proteins (e.g. gel filtration, affinity, and ion exchange), hydrophobic interaction chromatography (HIC) commonly requires experimental determination (referred to as screening or "scouting") in order to select the most suitable chromatographic medium for purifying a given protein 1. The method presented here describes an automated approach to scouting for an optimal HIC media to be used in protein purification. HIC separates proteins and other biomolecules from a crude lysate based on differences in hydrophobicity. Similar to affinity chromatography (AC) and ion exchange chromatography (IEX), HIC is capable of concentrating the protein of interest as it progresses through the chromatographic process. Proteins best suited for purification by HIC include those with hydrophobic surface regions and able to withstand exposure to salt concentrations in excess of 2 M ammonium sulfate ((NH4)2SO4). HIC is often chosen as a purification method for proteins lacking an affinity tag, and thus unsuitable for AC, and when IEX fails to provide adequate purification. Hydrophobic moieties on the protein surface temporarily bind to a nonpolar ligand coupled to an inert, immobile matrix. The interaction between protein and ligand are highly dependent on the salt concentration of the buffer flowing through the chromatography column, with high ionic concentrations strengthening the protein-ligand interaction and making the protein immobile (i.e. bound inside the column) 2. As salt concentrations decrease, the protein-ligand interaction dissipates, the protein again becomes mobile and elutes from the column. Several HIC media are commercially available in pre-packed columns, each containing one of several hydrophobic ligands (e.g. S-butyl, butyl, octyl, and phenyl) cross-linked at varying densities to agarose beads of a specific diameter 3. Automated column scouting allows for an efficient approach for determining which HIC media

  6. Supercritical fluid chromatography

    NASA Astrophysics Data System (ADS)

    Vigdergauz, M. S.; Lobachev, A. L.; Lobacheva, I. V.; Platonov, I. A.

    1992-03-01

    The characteristic features of supercritical fluid chromatography (SCFC) are examined and there is a brief historical note concerning the development of the method. Information concerning the use of supercritical fluid chromatography in the analysis of objects of different nature is presented in the form of a table. The roles of the mobile and stationary phases in the separation process and the characteristic features of the apparatus and of the use of the method in physicochemical research are discussed. The bibliography includes 364 references.

  7. Performance improvement of the one-dot lateral flow immunoassay for aflatoxin B1 by using a smartphone-based reading system.

    PubMed

    Lee, Sangdae; Kim, Giyoung; Moon, Jihea

    2013-04-18

    This study was conducted to develop a simple, rapid, and accurate lateral flow immunoassay (LFIA) detection method for point-of-care diagnosis. The one-dot LFIA for aflatoxin B1 (AFB1) was based on the modified competitive binding format using competition between AFB1 and colloidal gold-AFB1-BSA conjugate for antibody binding sites in the test zone. A Smartphone-based reading system consisting of a Samsung Galaxy S2 Smartphone, a LFIA reader, and a Smartphone application for the image acquisition and data analysis. The detection limit of one-dot LFIA for AFB1 is 5 μg/kg. This method provided semi-quantitative analysis of AFB1 samples in the range of 5 to 1,000 μg/kg. Using combination of the one-dot LFIA and the Smartphone-based reading system, it is possible to conduct a more fast and accurate point-of-care diagnosis.

  8. A summary of lateral-stability derivatives calculated for wing plan forms in supersonic flow

    NASA Technical Reports Server (NTRS)

    Jones, Arthur L; Alksne, Alberta

    1951-01-01

    A compilation of theoretical values of the lateral-stability derivatives for wings at supersonic speeds is presented in the form of design charts. The wing plan forms for which this compilation has been prepared include a rectangular, two trapezoidal, two triangular, a fully-tapered swept-back, a sweptback hexagonal, an unswept hexagonal, and a notched triangular plan form. A full set of results, that is, values for all nine of the lateral-stability derivatives for wings, was available for the first six of these plan forms only. The reasons for the incompleteness of the results available for other plan forms are discussed.

  9. Topographically Driven Lateral Water Fluxes and Their Influence on Carbon Assimilation of a Black Spruce Ecosystem.

    NASA Astrophysics Data System (ADS)

    Govind, A.; Chen, J. M.; Margolis, H.; Bernier, P. Y.

    2006-12-01

    Current estimates of ecophysiological indicators overlook the effects of topographically-driven lateral flow of soil water. We hypothesize that topographically driven lateral water flows over the landscape have significant influence on the terrestrial carbon cycle. To this end, we simulated the hydrological controls on carbon cycle processes in a black spruce forest in central Quebec, Canada, using the Boreal Ecosystem Productivity Simulator (BEPS) at a daily time step. We accounted for lateral surface and subsurface flows in BEPS by incorporating a distributed, process-oriented hydrological procedure. The results show that modeled dynamics of ecophysiological processes such as evapotranspiration (ET) and photosynthesis (GPP) are consistent with the spatial variation of land cover, topography, soil texture, and leaf area index. Simulated ET and GPP averaged within the footprint of an eddy covariance tower in the watershed agree well with flux measurements with R2=0.77 and 0.83 for ET and GPP, respectively. For ET simulation, much of the remaining discrepancies are found in the winter when the model underestimates snow sublimation. For GPP, there is an underestimation in the fall coinciding with a mid growing season drought, showing the high sensitivity of the model to the soil water status. The key processes controlling primary production were hydraulic limitations for water transfer from soil, roots, stems and leaves through stomatal conductance. Therefore, a further understanding of soil water dynamics is warranted. Comparison with the soil water content of the footprint- averaged unsaturated zone showed that the model captured the annual trend. We also simulated the variations in the water table as well as the mid growing season drought, with a reasonable accuracy(R2=0.68). The foot print average water budget reveals that the annual precipitation of 835mm is partitioned into 282mm of ET, 541 mm of subsurface runoff, and 6 mm of storage change. To test the

  10. Application of a SERS-based lateral flow immunoassay strip for the rapid and sensitive detection of staphylococcal enterotoxin B

    NASA Astrophysics Data System (ADS)

    Hwang, Joonki; Lee, Sangyeop; Choo, Jaebum

    2016-06-01

    A novel surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay (LFA) biosensor was developed to resolve problems associated with conventional LFA strips (e.g., limits in quantitative analysis and low sensitivity). In our SERS-based biosensor, Raman reporter-labeled hollow gold nanospheres (HGNs) were used as SERS detection probes instead of gold nanoparticles. With the proposed SERS-based LFA strip, the presence of a target antigen can be identified through a colour change in the test zone. Furthermore, highly sensitive quantitative evaluation is possible by measuring SERS signals from the test zone. To verify the feasibility of the SERS-based LFA strip platform, an immunoassay of staphylococcal enterotoxin B (SEB) was performed as a model reaction. The limit of detection (LOD) for SEB, as determined with the SERS-based LFA strip, was estimated to be 0.001 ng mL-1. This value is approximately three orders of magnitude more sensitive than that achieved with the corresponding ELISA-based method. The proposed SERS-based LFA strip sensor shows significant potential for the rapid and sensitive detection of target markers in a simplified manner.A novel surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay (LFA) biosensor was developed to resolve problems associated with conventional LFA strips (e.g., limits in quantitative analysis and low sensitivity). In our SERS-based biosensor, Raman reporter-labeled hollow gold nanospheres (HGNs) were used as SERS detection probes instead of gold nanoparticles. With the proposed SERS-based LFA strip, the presence of a target antigen can be identified through a colour change in the test zone. Furthermore, highly sensitive quantitative evaluation is possible by measuring SERS signals from the test zone. To verify the feasibility of the SERS-based LFA strip platform, an immunoassay of staphylococcal enterotoxin B (SEB) was performed as a model reaction. The limit of detection (LOD) for SEB, as

  11. A rapid Salmonella detection method involving thermophilic helicase-dependent amplification and a lateral flow assay.

    PubMed

    Du, Xin-Jun; Zhou, Tian-Jiao; Li, Ping; Wang, Shuo

    2017-08-01

    Salmonella is a major foodborne pathogen that is widespread in the environment and can cause serious human and animal disease. Since conventional culture methods to detect Salmonella are time-consuming and laborious, rapid and accurate techniques to detect this pathogen are critically important for food safety and diagnosing foodborne illness. In this study, we developed a rapid, simple and portable Salmonella detection strategy that combines thermophilic helicase-dependent amplification (tHDA) with a lateral flow assay to provide a detection result based on visual signals within 90 min. Performance analyses indicated that the method had detection limits for DNA and pure cultured bacteria of 73.4-80.7 fg and 35-40 CFU, respectively. Specificity analyses showed no cross reactions with Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Enterobacter aerogenes, Shigella and Campylobacter jejuni. The results for detection in real food samples showed that 1.3-1.9 CFU/g or 1.3-1.9 CFU/mL of Salmonella in contaminated chicken products and infant nutritional cereal could be detected after 2 h of enrichment. The same amount of Salmonella in contaminated milk could be detected after 4 h of enrichment. This tHDA-strip can be used for the rapid detection of Salmonella in food samples and is particularly suitable for use in areas with limited equipment. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Cerebral blood flow laterality derived from arterial spin labeling as a biomarker for assessing the disease severity of parkinson's disease.

    PubMed

    Yamashita, Koji; Hiwatashi, Akio; Togao, Osamu; Kikuchi, Kazufumi; Yamaguchi, Hiroo; Suzuki, Yuriko; Kamei, Ryotaro; Yamasaki, Ryo; Kira, Jun-Ichi; Honda, Hiroshi

    2017-06-01

    To evaluate cerebral blood flow (CBF) laterality derived from arterial spin labeling (ASL) in early-stage Parkinson's disease (PD) patients compared with those with advanced stages. Thirty-eight patients with PD (21 patients in early stages, 17 patients in advanced stages) were retrospectively studied. The CBF maps derived from 3T ASL data were co-registered to the corresponding 3DT1WI using SPM 12 software. Caudate nucleus (CN), putamen (PT), globus pallidus (GP), and thalamus (TH) were manually traced on the representative axial slices of 3DT1WI. CBF of the CN, PT, GP, and TH was measured using corresponding pixels on the co-registered CBF maps. A laterality index (LI) was calculated as the ratio of the contralateral CBF to primary affected side CBF. Each LI was compared between early and advanced stages of PD using the Mann-Whitney U-test. The LIs were also compared between each stage of PD. In the CN, the LIs were significantly higher in early stages (mean LI ± SD, 95% confidence interval = 1.06 ± 0.14, 1.00-1.13) than in advanced stages (0.94 ± 0.14, 0.87-1.01; P < 0.05). We also observed a tendency toward decreased LIs with disease severity (1.10 ± 0.14, 0.99-1.21 for Hoehn and Yahr stage I; 1.04 ± 0.14, 0.92-1.12 for stage II; 0.96 ± 0.11, 0.89-1.10 for stage III; 0.93 ± 0.17, 0.81-1.05 for stage IV). The evaluation of CBF laterality pattern in the CN using ASL may be useful for assessing the disease severity of PD patients. 3 J. MAGN. RESON. IMAGING 2017;45:1821-1826. © 2016 International Society for Magnetic Resonance in Medicine.

  13. Cross-axis synchronous flow-through coil planet centrifuge for large-scale preparative counter-current chromatography. III. Performance of large-bore coils in slow planetary motion.

    PubMed

    Ito, Y; Zhang, T Y

    1988-11-25

    A preparative capability of the present cross-axis synchronous flow-through coil planet centrifuge was demonstrated with 0.5 cm I.D. multilayer coils. Results of the model studies with short coils indicated that the optimal separations are obtained at low revolutional speeds of 100-200 rpm in both central and lateral coil positions. Preparative separations were successfully performed on 2.5-10 g quantities of test samples in a pair of multilayer coils connected in series with a total capacity of 2.5 l. The sample loading capacity will be scaled up in several folds by increasing the column width.

  14. Low-Cost 3-D Flow Estimation of Blood With Clutter.

    PubMed

    Wei, Siyuan; Yang, Ming; Zhou, Jian; Sampson, Richard; Kripfgans, Oliver D; Fowlkes, J Brian; Wenisch, Thomas F; Chakrabarti, Chaitali

    2017-05-01

    Volumetric flow rate estimation is an important ultrasound medical imaging modality that is used for diagnosing cardiovascular diseases. Flow rates are obtained by integrating velocity estimates over a cross-sectional plane. Speckle tracking is a promising approach that overcomes the angle dependency of traditional Doppler methods, but suffers from poor lateral resolution. Recent work improves lateral velocity estimation accuracy by reconstructing a synthetic lateral phase (SLP) signal. However, the estimation accuracy of such approaches is compromised by the presence of clutter. Eigen-based clutter filtering has been shown to be effective in removing the clutter signal; but it is computationally expensive, precluding its use at high volume rates. In this paper, we propose low-complexity schemes for both velocity estimation and clutter filtering. We use a two-tiered motion estimation scheme to combine the low complexity sum-of-absolute-difference and SLP methods to achieve subpixel lateral accuracy. We reduce the complexity of eigen-based clutter filtering by processing in subgroups and replacing singular value decomposition with less compute-intensive power iteration and subspace iteration methods. Finally, to improve flow rate estimation accuracy, we use kernel power weighting when integrating the velocity estimates. We evaluate our method for fast- and slow-moving clutter for beam-to-flow angles of 90° and 60° using Field II simulations, demonstrating high estimation accuracy across scenarios. For instance, for a beam-to-flow angle of 90° and fast-moving clutter, our estimation method provides a bias of -8.8% and standard deviation of 3.1% relative to the actual flow rate.

  15. Sample displacement chromatography as a method for purification of proteins and peptides from complex mixtures

    PubMed Central

    Gajdosik, Martina Srajer; Clifton, James; Josic, Djuro

    2012-01-01

    Sample displacement chromatography (SDC) in reversed-phase and ion-exchange modes was introduced approximately twenty years ago. This method takes advantage of relative binding affinities of components in a sample mixture. During loading, there is a competition among different sample components for the sorption on the surface of the stationary phase. SDC was first used for the preparative purification of proteins. Later, it was demonstrated that this kind of chromatography can also be performed in ion-exchange, affinity and hydrophobic-interaction mode. It has also been shown that SDC can be performed on monoliths and membrane-based supports in both analytical and preparative scale. Recently, SDC in ion-exchange and hydrophobic interaction mode was also employed successfully for the removal of trace proteins from monoclonal antibody preparations and for the enrichment of low abundance proteins from human plasma. In this review, the principals of SDC are introduced, and the potential for separation of proteins and peptides in micro-analytical, analytical and preparative scale is discussed. PMID:22520159

  16. Quasi-radial wall jets as a new concept in boundary layer flow control

    NASA Astrophysics Data System (ADS)

    Javadi, Khodayar; Hajipour, Majid

    2018-01-01

    This work aims to introduce a novel concept of wall jets wherein the flow is radially injected into a medium through a sector of a cylinder, called quasi-radial (QR) wall jets. The results revealed that fluid dynamics of the QR wall jet flow differs from that of conventional wall jets. Indeed, lateral and normal propagations of a conventional three-dimensional wall jet are via shear stresses. While, lateral propagation of a QR wall jet is due to mean lateral component of the velocity field. Moreover, discharged Arrays of conventional three-dimensional wall jets in quiescent air lead to formation of a combined wall jet at large distant from the nozzles, while QR wall jet immediately spread in lateral direction, meet each other and merge together very quickly in a short distance downstream of the jet nozzles. Furthermore, in discharging the conventional jets into an external flow, there is no strong interaction between them as they are moving parallel. While, in QR wall jets the lateral components of the velocity field strongly interact with boundary layer of the external flow and create strong helical vortices acting as vortex generators.

  17. Advanced proteomic liquid chromatography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Xie, Fang; Smith, Richard D.; Shen, Yufeng

    2012-10-26

    Liquid chromatography coupled with mass spectrometry is the predominant platform used to analyze proteomics samples consisting of large numbers of proteins and their proteolytic products (e.g., truncated polypeptides) and spanning a wide range of relative concentrations. This review provides an overview of advanced capillary liquid chromatography techniques and methodologies that greatly improve separation resolving power and proteomics analysis coverage, sensitivity, and throughput.

  18. Thin-Layer and Paper Chromatography.

    ERIC Educational Resources Information Center

    Sherma, Joseph; Fried, Bernard

    1984-01-01

    Reviews literature on chromatography examining: books, reviews, student experiments; chromatographic systems, techniques, apparatus; detecting and identification of separated zones; preparative chromatography and radiochromatography; and applications related to specific materials (such as acids, alcohols, amino acids, antibiotics, enzymes, dyes,…

  19. Concentration and fractionation of hydrophobic organic acid constituents from natural waters by liquid chromatography

    USGS Publications Warehouse

    Thurman, E.M.; Malcolm, R.L.

    1979-01-01

    A scheme is presented which used adsorption chromatography with pH gradient elution and size-exclusion chromatography to concentrate and separate hydrophobic organic acids from water. A review of chromatographic processes involved in the flow scheme is also presented. Organic analytes which appear in each aqueous fraction are quantified by dissolved organic carbon analysis. Hydrophobic organic acids in a water sample are concentrated on a porous acrylic resin. These acids usually constitute approximately 30-50 percent of the dissolved organic carbon in an unpolluted water sample and are eluted with an aqueous eluent (dilute base). The concentrate is then passed through a column of polyacryloylmorpholine gel, which separates the acids into high- and low-molecular-weight fractions. The high- and low-molecular-weight eluates are reconcentrated by adsorption chromatography, then are eluted with a pH gradient into strong acids (predominately carboxylic acids) and weak acids (predominately phenolic compounds). For standard compounds and samples of unpolluted waters, the scheme fractionates humic substances into strong and weak acid fractions that are separated from the low molecular weight acids. A new method utilizing conductivity is also presented to estimate the acidic components in the methanol fraction.

  20. Sample injector for high pressure liquid chromatography

    DOEpatents

    Paul, Phillip H.; Arnold, Don W.; Neyer, David W.

    2001-01-01

    Apparatus and method for driving a sample, having a well-defined volume, under pressure into a chromatography column. A conventional high pressure sampling valve is replaced by a sample injector composed of a pair of injector components connected in series to a common junction. The injector components are containers of porous dielectric material constructed so as to provide for electroosmotic flow of a sample into the junction. At an appropriate time, a pressure pulse from a high pressure source, that can be an electrokinetic pump, connected to the common junction, drives a portion of the sample, whose size is determined by the dead volume of the common junction, into the chromatographic column for subsequent separation and analysis. The apparatus can be fabricated on a substrate for microanalytical applications.

  1. Enantioseparation of napropamide by supercritical fluid chromatography: Effects of the chromatographic conditions and separation mechanism.

    PubMed

    Zhao, Lu; Xie, Jingqian; Guo, Fangjie; Liu, Kai

    2018-05-01

    Supercritical fluid chromatography (SFC) is already used for enantioseparation in the pharmaceutical industry, but it is rarely used for the separation of chiral pesticides. Comparing with high performence liquid chromatography, SFC uses much more environmnetal friendly and economic mobile phase, supercritical CO 2 . In our work, the enantioseparation of an amide herbicide, napropamide, using three different polysaccharide-type chiral stationary phases (CSPs) in SFC was investigated. By studying the effect of different CSPs, organic modifiers, temperature, back-pressure regulator pressures, and flow rates for the enantioseparation of napropamide, we established a rapid and green method for enantioseparation that takes less than 2 minutes: The column was CEL2, the mobile phase was CO 2 with 20% 2-propanol, and the flow rate was 2.0 mL/min. We found that CEL2 demonstrated the strongest resolution capability. Acetonitrile was favored over alcoholic solvents when the CSP was amylose and 2-propanol was the best choice when using cellulose. When the concentration of the modifiers or the flow rate was decreased, resolutions and analysis times increased concurrently. The temperature and back-pressure regulator pressure exhibited only minor influences on the resolution and analysis time of the napropamide enantioseparations with these chiral columns. The molecular docking analysis provided a deeper insight into the interactions between the enantiomers and the CSPs at the atomic level and partly explained the reason for the different elution orders using the different chiral columns. © 2018 Wiley Periodicals, Inc.

  2. Enabling Microliquid Chromatography by Microbead Packing of Microchannels

    NASA Technical Reports Server (NTRS)

    Balvin, Manuel; Zheng, Yun

    2013-01-01

    The microbead packing is the critical element required in the success of on-chip microfabrication of critical microfluidic components for in-situ analysis and detection of chiral amino acids. In order for microliquid chromatography to occur, there must be a stationary phase medium within the microchannel that interacts with the analytes present within flowing fluid. The stationary phase media are the microbeads packed by the process discussed in this work. The purpose of the microliquid chromatography is to provide a lightweight, low-volume, and low-power element to separate amino acids and their chiral partners efficiently to understand better the origin of life. In order to densely pack microbeads into the microchannels, a liquid slurry of microbeads was created. Microbeads were extracted from a commercially available high-performance liquid chromatography column. The silica beads extracted were 5 microns in diameter, and had surface coating of phenyl-hexyl. These microbeads were mixed with a 200- proof ethanol solution to create a microbead slurry with the right viscosity for packing. A microfilter is placed at the outlet via of the microchannel and the slurry is injected, then withdrawn across a filter using modified syringes. After each injection, the channel is flushed with ethanol to enhance packing. This cycle is repeated numerous times to allow for a tightly packed channel of microbeads. Typical microbead packing occurs in the macroscale into tubes or channels by using highly pressurized systems. Moreover, these channels are typically long and straight without any turns or curves. On the other hand, this method of microbead packing is completed within a microchannel 75 micrometers in diameter. Moreover, the microbead packing is completed into a serpentine type microchannel, such that it maximizes microchannel length within a microchip. Doing so enhances the interactions of the analytes with the microbeads to separate efficiently amino acids and amino acid

  3. Enabling Microliquid Chromatography by Microbead Packing of Microchannels

    NASA Technical Reports Server (NTRS)

    Balvin, Manuel; Zheng, Yun

    2014-01-01

    The microbead packing is the critical element required in the success of on-chip microfabrication of critical microfluidic components for in-situ analysis and detection of chiral amino acids. In order for microliquid chromatography to occur, there must be a stationary phase medium within the microchannel that interacts with the analytes present within flowing fluid. The stationary phase media are the microbeads packed by the process discussed in this work. The purpose of the microliquid chromatography is to provide a lightweight, low-volume, and low-power element to separate amino acids and their chiral partners efficiently to understand better the origin of life. In order to densely pack microbeads into the microchannels, a liquid slurry of microbeads was created. Microbeads were extracted from a commercially available high-performance liquid chromatography column. The silica beads extracted were 5 microns in diameter, and had surface coating of phenyl-hexyl. These microbeads were mixed with a 200- proof ethanol solution to create a microbead slurry with the right viscosity for packing. A microfilter is placed at the outlet via of the microchannel and the slurry is injected, then withdrawn across a filter using modified syringes. After each injection, the channel is flushed with ethanol to enhance packing. This cycle is repeated numerous times to allow for a tightly packed channel of microbeads. Typical microbead packing occurs in the macroscale into tubes or channels by using highly pressurized systems. Moreover, these channels are typically long and straight without any turns or curves. On the other hand, this method of microbead packing is completed within a microchannel 75 micrometers in diameter. Moreover, the microbead packing is completed into a serpentine type microchannel, such that it maximizes microchannel length within a microchip. Doing so enhances the interactions of the analytes with the microbeads to separate efficiently amino acids and amino acid

  4. Membrane-based lateral flow immunochromatographic strip with nanoparticles as reporters for detection: A review.

    PubMed

    Huang, Xiaolin; Aguilar, Zoraida P; Xu, Hengyi; Lai, Weihua; Xiong, Yonghua

    2016-01-15

    Membrane-based lateral flow immunochromatographic strip (LFICS) is widely used in various fields because of its simplicity, rapidity (detection within 10min), and low cost. However, early designs of membrane-based LFICS for preliminary screening only provide qualitative ("yes/no" signal) or semi-quantitative results without quantitative information. These designs often suffer from low-signal intensity and poor sensitivity and are only capable of single analyte detection, not simultaneous multiple detections. The performance of existing techniques used for detection using LFICS has been considerably improved by incorporating different kinds of nanoparticles (NPs) as reporters. NPs can serve as alternative labels and improve analytical sensitivity or limit of detection of LFICS because of their unique properties, such as optical absorption, fluorescence spectra, and magnetic properties. The controlled manipulation of NPs allows simultaneous or multiple detections by using membrane-based LFICS. In this review, we discuss how colored (e.g., colloidal gold, carbon, and colloidal selenium NPs), luminescent (e.g., quantum dots, up-converting phosphor NPs, and dye-doped NPs), and magnetic NPs are integrated into membrane-based LFICS for the detection of target analytes. Gold NPs are also featured because of their wide applications. Different types and unique properties of NPs are briefly explained. This review focuses on examples of NP-based LFICS to illustrate novel concepts in various devices with potential applications as screening tools. This review also highlights the superiority of NP-based approaches over existing conventional strategies for clinical analysis, food safety, and environmental monitoring. This paper is concluded by a short section on future research trends regarding NP-based LFICS. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Microfabrication of IPMC cilia for bio-inspired flow sensing

    NASA Astrophysics Data System (ADS)

    Lei, Hong; Li, Wen; Tan, Xiaobo

    2012-04-01

    As the primary flow sensing organ for fishes, the lateral line system plays a critical role in fish behavior. Analogous to its biological counterpart, an artificial lateral line system, consisting of arrays of micro flow sensors, is expected to be instrumental in the navigation and control of underwater robots. In this paper we investigate the microfabrication of ionic polymer-metal composite (IPMC) cilia for the purpose of flow sensing. While existing macro- and microfabrication methods for IPMCs have predominantly focused on planar structures, we propose a device where micro IPMC beams stand upright on a substrate to effectively interact with the flow. Challenges in the casting of 3D Nafion structure and selective formation of electrodes are discussed, and potential solutions for addressing these challenges are presented together with preliminary microfabrication results.

  6. Gas Chromatography.

    ERIC Educational Resources Information Center

    Cram, Stuart P.; And Others

    1980-01-01

    Selects fundamental developments in theory, methodology, and instrumentation in gas chromatography (GC). A special section reviews GC in the People's Republic of China. Over 1,000 references are cited. (CS)

  7. PrimaTB STAT-PAK Assay, a Novel, Rapid Lateral-Flow Test for Tuberculosis in Nonhuman Primates▿

    PubMed Central

    Lyashchenko, Konstantin P.; Greenwald, Rena; Esfandiari, Javan; Greenwald, David; Nacy, Carol A.; Gibson, Susan; Didier, Peter J.; Washington, Marc; Szczerba, Peter; Motzel, Sherri; Handt, Larry; Pollock, John M.; McNair, James; Andersen, Peter; Langermans, Jan A. M.; Verreck, Frank; Ervin, Sean; Ervin, Frank; McCombs, Candace

    2007-01-01

    Tuberculosis (TB) is the most important zoonotic bacterial disease in nonhuman primates (NHP). The current diagnostic method, the intradermal palpebral tuberculin test, has serious shortcomings. We characterized antibody responses in NHP against Mycobacterium tuberculosis to identify immunodominant antigens and develop a rapid serodiagnostic test for TB. A total of 422 NHP were evaluated, including 243 rhesus (Macaca mulatta), 46 cynomolgus (Macaca fascicularis), and 133 African green (Cercopithecus aethiops sabaeus) monkeys at five collaborative centers. Of those, 50 monkeys of the three species were experimentally inoculated with M. tuberculosis. Antibody responses were monitored every 2 to 4 weeks for up to 8 months postinfection by MultiAntigen Print ImmunoAssay with a panel of 12 recombinant antigens. All of the infected monkeys produced antibodies at various levels and with different antigen recognition patterns. ESAT-6 and MPB83 were the most frequently recognized proteins during infection. A combination of selected antigens which detected antibodies in all of the infected monkeys was designed to develop the PrimaTB STAT-PAK assay by lateral-flow technology. Serological evaluation demonstrated high diagnostic sensitivity (90%) and specificity (99%). The highest rate of TB detection was achieved when the skin test was combined with the PrimaTB STAT-PAK kit. This novel immunoassay provides a simple, rapid, and accurate test for TB in NHP. PMID:17652522

  8. Advanced proteomic liquid chromatography

    PubMed Central

    Xie, Fang; Smith, Richard D.; Shen, Yufeng

    2012-01-01

    Liquid chromatography coupled with mass spectrometry is the predominant platform used to analyze proteomics samples consisting of large numbers of proteins and their proteolytic products (e.g., truncated polypeptides) and spanning a wide range of relative concentrations. This review provides an overview of advanced capillary liquid chromatography techniques and methodologies that greatly improve separation resolving power and proteomics analysis coverage, sensitivity, and throughput. PMID:22840822

  9. Preparation, characterization and application of a reversed phase liquid chromatography/hydrophilic interaction chromatography mixed-mode C18-DTT stationary phase.

    PubMed

    Wang, Qing; Long, Yao; Yao, Lin; Xu, Li; Shi, Zhi-Guo; Xu, Lanying

    2016-01-01

    A mixed-mode chromatographic stationary phase, C18-DTT (dithiothreitol) silica (SiO2) was prepared through "thiol-ene" click chemistry. The obtained material was characterized by fourier transform infrared spectroscope, nitrogen adsorption analysis and contact angle analysis. Chromatographic performance of the C18-DTT was systemically evaluated by studying the effect of acetonitrile content, pH, buffer concentration of the mobile phase and column temperature. It was demonstrated that the novel stationary phase possessed reversed phase liquid chromatography (RPLC)/hydrophilic interaction liquid chromatography (HILIC) mixed-mode property. The stop-flow test revealed that C18-DTT exhibited excellent compatibility with 100% aqueous mobile phase. Additionally, the stability and column-to-column reproducibility of the C18-DTT material were satisfactory, with relative standard deviations of retention factor of the tested analytes (verapamil, fenbufen, guanine, tetrandrine and nicotinic acid) in the range of 1.82-3.72% and 0.85-1.93%, respectively. Finally, the application of C18-DTT column was demonstrated in the separation of non-steroidal anti-inflammatory drugs, aromatic carboxylic acids, alkaloids, nucleo-analytes and polycyclic aromatic hydrocarbons. It had great resolving power in the analysis of various compounds in HILIC and RPLC chromatographic conditions and was a promising RPLC/HILIC mixed-mode stationary phase. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Study of the influence of surfactants on the activity coefficients and mass transfer coefficients of methanol in aqueous mixtures by reversed-flow gas chromatography.

    PubMed

    Kotsalos, Efthimios; Brezovska, Boryana; Sevastos, Dimitrios; Vagena, Artemis; Koliadima, Athanasia; Kapolos, John; Karaiskakis, George

    2017-11-17

    This work focuses on the influences of surfactants on the activity coefficients, γ, of methanol in binary mixtures with water, as well as on the mass transfer coefficients, k c , for the evaporation of methanol, which is a ubiquitous component in the troposphere, from mixtures of methanol with water at various surfactant's and methanol's concentrations. The technique used is the Reversed-Flow Gas Chromatography (R.F.G.C.), a version of Inverse Gas Chromatography, which allows determining both parameters by performing only one experiment for the k c parameter and two experiments for the γ parameter. The k c and γ values decrease in the presence of the three surfactants used (CTAB, SDS, TRITON X-100) at all methanol's and surfactant's concentrations. The decrease in the methanol's molar fraction, at constant number of surfactant films leads to a decrease in the k c and γ values, while the decrease in the surfactant's concentration, at constant methanol's molar fraction leads to an increase in both the k c and γ parameters. Mass transfer coefficients for the evaporation of methanol at the surfactant films, are also calculated which are approximately between 4 and 5 orders of magnitude larger than the corresponding mass transfer coefficients at the liquid films. Finally, thicknesses of the boundary layer of methanol in the mixtures of methanol with water were determined. The quantities found are compared with those given in the literature or calculated theoretically using various empirical equations. The precision of the R.F.G.C. method for measuring γ and k c parameters is approximately high (94.3-98.0%), showing that R.F.G.C. can be used with success not only for the thermodynamic study of solutions, but also for the interphase transport. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Constant pressure mode extended simple gradient liquid chromatography system for micro and nanocolumns.

    PubMed

    Šesták, Jozef; Kahle, Vladislav

    2014-07-11

    Performing gradient liquid chromatography at constant pressure instead of constant flow rate has serious potential for shortening the analysis time and increasing the productivity of HPLC instruments that use gradient methods. However, in the constant pressure mode the decreasing column permeability during a long period of time negatively affects the repeatability of retention time. Thus a volume-based approach, in which the detector signal is plotted as a function of retention volume, must be taken into consideration. Traditional HPLC equipment, however, requires quite complex hardware and software modifications in order to work at constant pressure and in the volume-based mode. In this short communication, a low cost and easily feasible pressure-controlled extension of the previously described simple gradient liquid chromatography platform is proposed. A test mixture of four nitro esters was separated by 10-60% (v/v) acetone/water gradient and a high repeatability of retention volumes at 20MPa (RSD less than 0.45%) was realized. Separations were also performed at different values of pressure (20, 25, and 31MPa), and only small variations of the retention volumes (up to 0.8%) were observed. In this particular case, the gain in the analysis speed of 7% compared to the constant flow mode was realized at a constant pressure. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Very high pressure liquid chromatography using fully porous particles: quantitative analysis of fast gradient separations without post-run times.

    PubMed

    Stankovich, Joseph J; Gritti, Fabrice; Stevenson, Paul G; Beaver, Lois Ann; Guiochon, Georges

    2014-01-10

    Using a column packed with fully porous particles, four methods for controlling the flow rates at which gradient elution runs are conducted in very high pressure liquid chromatography (VHPLC) were tested to determine whether reproducible thermal conditions could be achieved, such that subsequent analyses would proceed at nearly the same initial temperature. In VHPLC high flow rates are achieved, producing fast analyses but requiring high inlet pressures. The combination of high flow rates and high inlet pressures generates local heat, leading to temperature changes in the column. Usually in this case a post-run time is input into the analytical method to allow the return of the column temperature to its initial state. An alternative strategy involves operating the column without a post-run equilibration period and maintaining constant temperature variations for subsequent analysis after conducting one or a few separations to bring the column to a reproducible starting temperature. A liquid chromatography instrument equipped with a pressure controller was used to perform constant pressure and constant flow rate VHPLC separations. Six replicate gradient separations of a nine component mixture consisting of acetophenone, propiophenone, butyrophenone, valerophenone, hexanophenone, heptanophenone, octanophenone, benzophenone, and acetanilide dissolved in water/acetonitrile (65:35, v/v) were performed under various experimental conditions: constant flow rate, two sets of constant pressure, and constant pressure operation with a programmed flow rate. The relative standard deviations of the response factors for all the analytes are lower than 5% across the methods. Programming the flow rate to maintain a fairly constant pressure instead of using instrument controlled constant pressure improves the reproducibility of the retention times by a factor of 5, when plotting the chromatograms in time. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Performance Improvement of the One-Dot Lateral Flow Immunoassay for Aflatoxin B1 by Using a Smartphone-Based Reading System

    PubMed Central

    Lee, Sangdae; Kim, Giyoung; Moon, Jihea

    2013-01-01

    This study was conducted to develop a simple, rapid, and accurate lateral flow immunoassay (LFIA) detection method for point-of-care diagnosis. The one-dot LFIA for aflatoxin B1 (AFB1) was based on the modified competitive binding format using competition between AFB1 and colloidal gold-AFB1-BSA conjugate for antibody binding sites in the test zone. A Smartphone-based reading system consisting of a Samsung Galaxy S2 Smartphone, a LFIA reader, and a Smartphone application for the image acquisition and data analysis. The detection limit of one-dot LFIA for AFB1 is 5 μg/kg. This method provided semi-quantitative analysis of AFB1 samples in the range of 5 to 1,000 μg/kg. Using combination of the one-dot LFIA and the Smartphone-based reading system, it is possible to conduct a more fast and accurate point-of-care diagnosis. PMID:23598499

  14. Preparative chromatography for specific δ13C isotopic analysis of individual carbohydrates in environmental samples

    NASA Astrophysics Data System (ADS)

    Nouara, Amel; Panagiotopoulos, Christos; Balesdent, Jérôme; Sempéré, Richard

    2017-04-01

    Carbohydrates are among the most abundant organic molecules on the Earth and are present in all geochemical systems. Despite their high abundance in the environment, very few studies assessed their origin using molecular carbohydrate isotopic analyses. In contrast with bulk stable isotope analysis (BSIA), which gives the isotopic signature of the entire sample without any specification about its chemical composition, compound specific 13C isotopic analysis of individual sugars (CSIA) offers valuable information about the origin of single molecules. Previous investigations used gas or liquid chromatography coupled with isotope ratio mass spectroscopy (GC-IRMS; HPLC-IRMS) for CSIA of sugars however the former requires δ13C corrections due to the carbon added to the sugar (derivatization) while the later does not provide always adequate separations among monosaccharides. Here we used cation preparative chromatography (Ca2+, Pb2+ and Na+) with refractive index detection in order to produce pure monosaccharide targets for subsequent EA-IRMS analyses. Milli-Q water was used as eluant at a flow rate 0.6 ml min-1. In general, three successive purifications (Ca2+, Pb2+, Ca2+) were sufficient to produce pure compounds. Pure monosaccharides were compared with authentic monosaccharide standards using 1H NMR and/or mass spectroscopy. The detection limit of our technique was about 1µM/sugar with a precision of 10% (n=6). Blanks run with Milli-Q water after three successive purifications resulted in carbon content of 0.13 to 2.77 µgC per collected sugar. These values are much lower than the minimum required amount (5 µgC) of the EA-IRSMS system with a precision of ± 0.35 ‰. Application of our method to environmental samples resulted in δ13C values of glucose, fructose, and levoglucosan in the range of -24 to -26 ‰ (PM10 atmospheric particles), and -15‰ to -22 ‰ for arabinose, glucose, and xylose (marine high molecular dissolved organic matter). These results fall in

  15. Experimental Demonstration of 3-Dimensional Flow Structures and Depositional Features in a Lateral Recirculation Zone

    NASA Astrophysics Data System (ADS)

    Grams, P. E.; Schmeeckle, M. W.; Mueller, E. R.; Buscombe, D.; Kasprak, A.; Leary, K. P.

    2016-12-01

    The connections between stream hydraulics, geomorphology and ecosystems in mountain rivers have been substantially perturbed by humans, for example through flow regulation related to hydropower activities. It is well known that the ecosystem impacts downstream of hydropower dams may be managed by a properly designed compensation release or environmental flows ("e-flows"), and such flows may also include sediment considerations (e.g. to break up bed armor). However, there has been much less attention given to the ecosystem impacts of water intakes (where water is extracted and transferred for storage and/or power production), even though in many mountain systems such intakes may be prevalent. Flow intakes tend to be smaller than dams and because they fill quickly in the presence of sediment delivery, they often need to be flushed, many times within a day in Alpine glaciated catchments with high sediment yields. The associated short duration "flood" flow is characterised by very high sediment concentrations, which may drastically modify downstream habitat, both during the floods but also due to subsequent accumulation of "legacy" sediment. The impacts on flora and fauna of these systems have not been well studied. In addition, there are no guidelines established that might allow the design of "e-flows" that also treat this sediment problem, something we call "sed-flows". Through an Alpine field example, we quantify the hydrological, geomorphological, and ecosystem impacts of Alpine water transfer systems. The high sediment concentrations of these flushing flows lead to very high rates of channel disturbance downstream, superimposed upon long-term and progressive bed sediment accumulation. Monthly macroinvertebrate surveys over almost a two-year period showed that reductions in the flushing rate reduced rates of disturbance substantially, and led to rapid macroinvertebrate recovery, even in the seasons (autumn and winter) when biological activity should be reduced

  16. Efficient separation of curcumin, demethoxycurcumin, and bisdemethoxycurcumin from turmeric using supercritical fluid chromatography: From analytical to preparative scale.

    PubMed

    Song, Wei; Qiao, Xue; Liang, Wen-fei; Ji, Shuai; Yang, Lu; Wang, Yuan; Xu, Yong-wei; Yang, Ying; Guo, De-an; Ye, Min

    2015-10-01

    Curcumin is the major constituent of turmeric (Curcuma longa L.). It has attracted widespread attention for its anticancer and anti-inflammatory activities. The separation of curcumin and its two close analogs, demethoxycurcumin and bisdemethoxycurcumin, has been challenging by conventional techniques. In this study, an environmentally friendly method based on supercritical fluid chromatography was established for the rapid and facile separation of the three curcuminoids directly from the methanol extract of turmeric. The method was first developed and optimized by ultra performance convergence chromatography, and was then scaled up to preparative supercritical fluid chromatography. Eluted with supercritical fluid CO2 containing 8-15% methanol (containing 10 mM oxalic acid) at a flow rate of 80 mL/min, curcumin, demethoxycurcumin and bisdemethoxycurcumin could be well separated on a Viridis BEH OBD column (Waters, 250 mm × 19 mm, 5 μm) within 6.5 min. As a result, 20.8 mg of curcumin (97.9% purity), 7.0 mg of demethoxycurcumin (91.1%), and 4.6 mg of bisdemethoxycurcumin (94.8%) were obtained after a single step of supercritical fluid chromatography separation with a mean recovery of 76.6%. Showing obvious advantages in low solvent consumption, large sample loading, and easy solvent removal, supercritical fluid chromatography was proved to be a superior technique for the efficient separation of natural products. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Characterization of synthetic dyes by comprehensive two-dimensional liquid chromatography combining ion-exchange chromatography and fast ion-pair reversed-phase chromatography.

    PubMed

    Pirok, Bob W J; Knip, Jitske; van Bommel, Maarten R; Schoenmakers, Peter J

    2016-03-04

    In the late 19th century, newly invented synthetic dyes rapidly replaced the natural dyes on the market. The characterization of mixtures of these so-called early synthetic dyes is complicated through the occurrence of many impurities and degradation products. Conventional one-dimensional liquid chromatography does not suffice to obtain fingerprints with sufficient resolution and baseline integrity. Comprehensive two-dimensional liquid chromatography (LC×LC) is employed in this study, with ion-exchange chromatography in the first dimension and fast ion-pair liquid chromatography in the second. Retention in the first dimension is largely determined by the number of charges, while the selection of a small ion-pair reagent (tetramethylammonium hydroxide) in the second dimension causes retention to be largely determined by the molecular structure of the dye. As a result, there is a high degree of orthogonality of the two dimensions, similar to the values typically encountered in GC×GC. The proposed LC×LC method shows a theroretical peak capacity of about 2000 in an analysis time of about three hours. Clear, informative fingerprints are obtained that open a way to a more efficient characterization of dyes used in objects of cultural heritage. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Evaluation between ultrahigh pressure liquid chromatography and high-performance liquid chromatography analytical methods for characterizing natural dyestuffs.

    PubMed

    Serrano, Ana; van Bommel, Maarten; Hallett, Jessica

    2013-11-29

    An evaluation was undertaken of ultrahigh pressure liquid chromatography (UHPLC) in comparison to high-performance liquid chromatography (HPLC) for characterizing natural dyes in cultural heritage objects. A new UHPLC method was optimized by testing several analytical parameters adapted from prior UHPLC studies developed in diverse fields of research. Different gradient elution programs were tested on seven UHPLC columns with different dimensions and stationary phase compositions by applying several mobile phases, flow rates, temperatures, and runtimes. The UHPLC method successfully provided more improved data than that achieved by the HPLC method. Indeed, even though carminic acid has shown circa 146% higher resolution with HPLC, UHPLC resulted in an increase of 41-61% resolution and a decrease of 91-422% limit of detection, depending on the dye compound. The optimized method was subsequently assigned to analyse 59 natural reference materials, in which 85 different components were ascribed with different physicochemical properties, in order to create a spectral database for future characterization of dyes in cultural heritage objects. The majority of these reference samples could be successfully distinguished with one single method through the examination of these compounds' retention times and their spectra acquired with a photodiode array detector. These results demonstrate that UHPLC analyses are extremely valuable for the acquisition of more precise chromatographic information concerning natural dyes with complex mixtures of different and/or closely related physicochemical properties, essential for distinguishing similar species of plants and animals used to colour cultural heritage objects. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Neonatal cytokines and cerebral palsy in very preterm infants.

    PubMed

    Nelson, Karin B; Grether, Judith K; Dambrosia, James M; Walsh, Eileen; Kohler, Shawn; Satyanarayana, Gowri; Nelson, Phillip G; Dickens, Benjamin F; Phillips, Terry M

    2003-04-01

    To examine the relationship of cytokines in blood of very preterm neonates with later diagnosis of spastic cerebral palsy (CP) compared with infants of similar gestational age without CP, we measured concentrations of inflammatory cytokines and other substances in archived neonatal blood by recycling immunoaffinity chromatography. Subjects were surviving children born before 32 wk gestational age (GA) to women without preeclampsia, 64 with later diagnoses of CP and 107 control children. The initial analyses were augmented by measurement of 11 cytokines by a bead-based flow analytic system (Luminex) in an additional 37 children with CP and 34 control children from the same cohort. Concentrations of examined substances did not differ by presence of indicators of infection in mother, infant, or placenta. On ANOVA, concentrations of a number of cytokines were significantly related to neonatal ultrasound abnormalities (periventricular leukomalacia, ventricular enlargement, or moderate or severe germinal matrix hemorrhage). None of the substances measured either by immunoaffinity chromatography or flow analytic methods, including IL-1, -6, and -8 and tumor necrosis factor-alpha, was related to later diagnosis of CP or its subtypes. Inflammatory cytokines in neonatal blood of very premature infants did not distinguish those with later diagnoses of CP from control children.

  20. Improved assay to detect Plasmodium falciparum using an uninterrupted, semi-nested PCR and quantitative lateral flow analysis

    PubMed Central

    2013-01-01

    Background A rapid, non-invasive, and inexpensive point-of-care (POC) diagnostic for malaria followed by therapeutic intervention would improve the ability to control infection in endemic areas. Methods A semi-nested PCR amplification protocol is described for quantitative detection of Plasmodium falciparum and is compared to a traditional nested PCR. The approach uses primers that target the P. falciparum dihydrofolate reductase gene. Results This study demonstrates that it is possible to perform an uninterrupted, asymmetric, semi-nested PCR assay with reduced assay time to detect P. falciparum without compromising the sensitivity and specificity of the assay using saliva as a testing matrix. Conclusions The development of this PCR allows nucleic acid amplification without the need to transfer amplicon from the first PCR step to a second reaction tube with nested primers, thus reducing both the chance of contamination and the time for analysis to < two hours. Analysis of the PCR amplicon yield was adapted to lateral flow detection using the quantitative up-converting phosphor (UCP) reporter technology. This approach provides a basis for migration of the assay to a POC microfluidic format. In addition the assay was successfully evaluated with oral samples. Oral fluid collection provides a simple non-invasive method to collect clinical samples. PMID:23433252

  1. Rapid and visual detection of Mycobacterium tuberculosis complex using recombinase polymerase amplification combined with lateral flow strips.

    PubMed

    Ma, Qinglin; Liu, Houming; Ye, Feidi; Xiang, Guangxin; Shan, Wanshui; Xing, Wanli

    2017-12-01

    To definitively diagnose active pulmonary Tuberculosis (TB), Mycobacterium tuberculosis complex (MTBC) bacilli must be identified within clinical specimens from patients. In this study, we introduced a rapid and visual detection method of MTBC using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strips. The LF-RPA assay, read results with naked eyes, could detect as few as 5 genome copies of M. tuberculosis H37Rv (ATCC 27294) per reaction and had no cross-reactions with other control bacteria even using excessive amount of template DNA. The system could work well at a broad range of temperature 25-45 °C and reach detectable level even within 5 min. When testing a total of 137 clinical specimens, the sensitivity and specificity of the LF-RPA assay were 100% (95% CI: 95.94%-100%) and 97.92% (95% CI: 88.93%-99.95%), respectively, compared to culture identification method. Therefore, the LF-RPA system we have demonstrated is a rapid, simple, robust method for MTBC detection which, subject to the availability of a suitable sample extraction method, has the potentiality to diagnose TB at the point-of-care testing. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Exploratory laboratory study of lateral turbulent diffusion at the surface of an alluvial channel

    USGS Publications Warehouse

    Sayre, William W.; Chamberlain, A.R.

    1964-01-01

    In natural streams turbulent diffusion is one of the principal mechanisms by which liquid and suspended-particulate contaminants are dispersed in the flow. A knowledge of turbulence characteristics is therefore essential in predicting the dispersal rates of contaminants in streams. In this study the theory of diffusion by continuous movements for homogeneous turbulence is applied to lateral diffusion at the surface of an open channel in which there is uniform flow. An exploratory-laboratory investigation was conducted in which the lateral dispersion at the water surface of a sand-Led flume was studied by measuring the lateral spread from a point source of small floating polyethylene articles. The experiment was restricted to a single set of low and channel geometry conditions. The results of the study indicate that with certain restrictions lateral dispersion in alluvial channels may be successfully described by the theory of diffusion by continuous movements. The experiment demonstrates a means for evaluating the lateral diffusion coefficient and also methods for quantitatively estimating fundamental turbulence properties, such as the intensity and the Lagrangian integral scale of turbulence in an alluvial channel. The experimental results show that with increasing distance from the source the coefficient of lateral turbulent diffusion increases initially but tends toward a constant limiting value. This result is in accordance with turbulent diffusion theory. Indications are that the distance downstream from the source required for the diffusion coefficient to reach its limiting value is actually very small when compared to the length scale of most diffusion phenomena in natural streams which are of practical interest.

  3. Lateral and vertical distribution of downstream migrating juvenile sea lamprey

    USGS Publications Warehouse

    Sotola, V. Alex; Miehls, Scott M.; Simard, Lee G.; Marsden, J. Ellen

    2018-01-01

    Sea lamprey is considered an invasive and nuisance species in the Laurentian Great Lakes, Lake Champlain, and the Finger Lakes of New York and is a major focus of control efforts. Currently, management practices focus on limiting the area of infestation using barriers to block migratory adults, and lampricides to kill ammocoetes in infested tributaries. No control efforts currently target the downstream-migrating post-metamorphic life stage which could provide another management opportunity. In order to apply control methods to this life stage, a better understanding of their downstream movement patterns is needed. To quantify spatial distribution of downstream migrants, we deployed fyke and drift nets laterally and vertically across the stream channel in two tributaries of Lake Champlain. Sea lamprey was not randomly distributed across the stream width and lateral distribution showed a significant association with discharge. Results indicated that juvenile sea lamprey is most likely to be present in the thalweg and at midwater depths of the stream channel. Further, a majority of the catch occurred during high flow events, suggesting an increase in downstream movement activity when water levels are higher than base flow. Discharge and flow are strong predictors of the distribution of out-migrating sea lamprey, thus managers will need to either target capture efforts in high discharge areas of streams or develop means to guide sea lamprey away from these areas.

  4. Mixed-mode chromatography/isotope ratio mass spectrometry.

    PubMed

    McCullagh, James S O

    2010-03-15

    Liquid chromatography coupled to molecular mass spectrometry (LC/MS) has been a standard technique since the early 1970s but liquid chromatography coupled to high-precision isotope ratio mass spectrometry (LC/IRMS) has only been available commercially since 2004. This development has, for the first time, enabled natural abundance and low enrichment delta(13)C measurements to be applied to individual analytes in aqueous mixtures creating new opportunities for IRMS applications, particularly for the isotopic study of biological molecules. A growing number of applications have been published in a range of areas including amino acid metabolism, carbohydrates studies, quantification of cellular and plasma metabolites, dietary tracer and nucleic acid studies. There is strong potential to extend these to new compounds and complex matrices but several challenges face the development of LC/IRMS methods. To achieve accurate isotopic measurements, HPLC separations must provide baseline-resolution between analyte peaks; however, the design of current liquid interfaces places severe restrictions on compatible flow rates and in particular mobile phase compositions. These create a significant challenge on which reports associated with LC/IRMS have not previously focused. Accordingly, this paper will address aspects of chromatography in the context of LC/IRMS, in particular focusing on mixed-mode separations and their benefits in light of these restrictions. It aims to provide an overview of mixed-mode stationary phases and of ways to improve high aqueous separations through manipulation of parameters such as column length, temperature and mobile phase pH. The results of several practical experiments are given using proteogenic amino acids and nucleosides both of which are of noted importance in the LC/IRMS literature. This communication aims to demonstrate that mixed-mode stationary phases provide a flexible approach given the constraints of LC/IRMS interface design and acts as a

  5. Evaluating two process scale chromatography column header designs using CFD.

    PubMed

    Johnson, Chris; Natarajan, Venkatesh; Antoniou, Chris

    2014-01-01

    Chromatography is an indispensable unit operation in the downstream processing of biomolecules. Scaling of chromatographic operations typically involves a significant increase in the column diameter. At this scale, the flow distribution within a packed bed could be severely affected by the distributor design in process scale columns. Different vendors offer process scale columns with varying design features. The effect of these design features on the flow distribution in packed beds and the resultant effect on column efficiency and cleanability needs to be properly understood in order to prevent unpleasant surprises on scale-up. Computational Fluid Dynamics (CFD) provides a cost-effective means to explore the effect of various distributor designs on process scale performance. In this work, we present a CFD tool that was developed and validated against experimental dye traces and tracer injections. Subsequently, the tool was employed to compare and contrast two commercially available header designs. © 2014 American Institute of Chemical Engineers.

  6. Lateral-Line Detection of Underwater Objects: From Goldfish to Submarines

    NASA Astrophysics Data System (ADS)

    van Hemmen, J. Leo

    2010-03-01

    Fish and some aquatic amphibians use their mechanosensory lateral-line system to navigate by means of hydrodynamic cues. How a fish determines an object's position and shape only through the lateral-line system and the ensuing neuronal processing is still a challenging problem. Our studies have shown that both stimulus position and stimulus form can be determined within the range of about one fish length and are encoded through the response of the afferent nerves originating from the detectors. A minimal detection model of a vibrating sphere (a dipole) has now been extended to other stimuli such as translating spheres, ellipsoids, or even wakes (vortex rings). The theoretical model is fully verified by experimental data. We have also constructed an underwater robot with an artificial lateral-line system designed to detect e.g. the presence of walls by measuring the change of water flow around the body. We will show how a simple model fits experimental results obtained from trout and goldfish and how a submarine may well be able to detect underwater objects by using an artificial lateral-line system.

  7. Separation of four flavonol glycosides from Solanum rostratum Dunal using aqueous two-phase flotation followed by preparative high-performance liquid chromatography.

    PubMed

    Chang, Lin; Shao, Qian; Xi, Xingjun; Chu, Qiao; Wei, Yun

    2017-02-01

    Aqueous two-phase flotation followed by preparative high-performance liquid chromatography was used to separate four flavonol glycosides from Solanum rostratum Dunal. In the aqueous two-phase flotation section, the effects of sublation solvent, solution pH, (NH 4 ) 2 SO 4 concentration in aqueous solution, cosolvent, N 2 flow rate, flotation time, and volumes of the polyethylene glycol phase on the recovery were investigated in detail, and the optimal conditions were selected: 50 wt% polyethylene glycol 1000 ethanol solvent as the flotation solvent, pH 4, 350 g/L of (NH 4 ) 2 SO 4 concentration in aqueous phase, 40 mL/min of N 2 flow rate, 30 min of flotation time, 10.0 mL of flotation solvent volume, and two times. After aqueous two-phase flotation concentration, the flotation products were purified by preparative high-performance liquid chromatography. The purities of the final products A and B were 98.1 and 99.0%. Product B was the mixture of three compounds based on the analysis of high-performance liquid chromatography at the temperature of 10°C, while product A was hyperoside after the identification by nuclear magnetic resonance. Astragalin, 3'-O-methylquercetin 3-O-β-d-galactopyranoside, and 3'-O-methylquercetin 3-O-β-d-glucopyranoside were obtained with the purity of 93.8, 97.1, and 99.2%, respectively, after the further separation of product B using preparative high-performance liquid chromatography. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Breakdown of Burton Prim Slichter approach and lateral solute segregation in radially converging flows

    NASA Astrophysics Data System (ADS)

    Priede, J.; Gerbeth, G.

    2005-11-01

    A theoretical study is presented of the effect of a radially converging melt flow, which is directed away from the solidification front, on the radial solute segregation in simple solidification models. We show that the classical Burton-Prim-Slichter (BPS) solution describing the effect of a diverging flow on the solute incorporation into the solidifying material breaks down for the flows converging along the solidification front. The breakdown is caused by a divergence of the integral defining the effective boundary layer thickness which is the basic concept of the BPS theory. Although such a divergence can formally be avoided by restricting the axial extension of the melt to a layer of finite height, radially uniform solute distributions are possible only for weak melt flows with an axial velocity away from the solidification front comparable to the growth rate. There is a critical melt velocity for each growth rate at which the solution passes through a singularity and becomes physically inconsistent for stronger melt flows. To resolve these inconsistencies we consider a solidification front presented by a disk of finite radius R0 subject to a strong converging melt flow and obtain an analytic solution showing that the radial solute concentration depends on the radius r as ˜ln(R0/r) and ˜ln(R0/r) close to the rim and at large distances from it. The logarithmic increase of concentration is limited in the vicinity of the symmetry axis by the diffusion becoming effective at a distance comparable to the characteristic thickness of the solute boundary layer. The converging flow causes a solute pile-up forming a logarithmic concentration peak at the symmetry axis which might be an undesirable feature for crystal growth processes.

  9. [Separation and identification of 5 glycosidic flavor precursors in tobacco by ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry].

    PubMed

    Wu, Xinhua; Zhu, Ruizhi; Ren, Zhuoying; Wang, Kai; Mou, Dingrong; Wei, Wanzhi; Miao, Mingming

    2009-11-01

    A qualitative method for the identification of 5 main glycosidic flavor precursors in tobacco was developed by using ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI MS/MS) and gas chromatography-mass spectrometry (GC-MS). The glycosidic flavor precursors in tobacco were extracted with methanol, cleaned up with an XAD-2 column. The aglycones were later released by enzyme-mediated hydrolysis under the condition of pH 5. The 5 volatile aglycone moieties were identified by GC-MS standard spectra library. The precursor ions of glycosides were determined by using electrospray ionization mass spectrometry in negative ion mode, then the 5 glycosidic flavor precursors were identified by using product ion scan (MS2) finally, using UPLC-ESI MS/MS, separation and identification of 5 glycosidic flavor precursors were accomplished on an RP-C,8 column in the multiple reaction monitoring (MRM) mode by using methanol and acetic acid-ammonium acetate aqueous solution as eluent. This work lays a foundation for the analysis of glycosidic flavor precursors without the standards by using liquid chromatography-mass spectrometry.

  10. Evaluation of a direct injection nebulizer interface for flow injection analysis and high performance liquid chromatography with inductively coupled plasma-atomic emission spectroscopic detection

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    LaFreniere, K.E.

    A direct injection nebulizer (DIN) was designed, developed, and evaluated to determine its potential utilization as an effective interface for flow injection analysis (FIA) and high performance liquid chromatography (HPLC) coupled with inductively coupled plasma-atomic emission spectroscopic detection. The analytical figures of merit for the DIN when used as an interface for FIA-ICP-AES were found to be comparable to or better than those obtained with conventional pneumatic nebulization in terms of limits of detection (LODs), reproducibility, linearity, and interelement effects. Stable plasma operation was maintained for the DIN sample introduction of a variety of pure organic solvents, including acetonitrile, methanol,more » methylisobutylketone, and pyridine. The HPLC-DIN-ICP-AES facility was specifically applied for the speciation of inorganic and organometallic species contained in synthetic mixtures, vanilla extracts, and a variety of energy-related materials, such as shale oil process water, coal extracts, shale oil, crude oil, and an SRC II. Suggestions for future research are also considered. 227 refs., 44 figs., 15 tabs.« less

  11. Platinum Nanocatalyst Amplification: Redefining the Gold Standard for Lateral Flow Immunoassays with Ultrabroad Dynamic Range

    PubMed Central

    2017-01-01

    Paper-based lateral flow immunoassays (LFIAs) are one of the most widely used point-of-care (PoC) devices; however, their application in early disease diagnostics is often limited due to insufficient sensitivity for the requisite sample sizes and the short time frames of PoC testing. To address this, we developed a serum-stable, nanoparticle catalyst-labeled LFIA with a sensitivity surpassing that of both current commercial and published sensitivities for paper-based detection of p24, one of the earliest and most conserved biomarkers of HIV. We report the synthesis and characterization of porous platinum core–shell nanocatalysts (PtNCs), which show high catalytic activity when exposed to complex human blood serum samples. We explored the application of antibody-functionalized PtNCs with strategically and orthogonally modified nanobodies with high affinity and specificity toward p24 and established the key larger nanoparticle size regimes needed for efficient amplification and performance in LFIA. Harnessing the catalytic amplification of PtNCs enabled naked-eye detection of p24 spiked into sera in the low femtomolar range (ca. 0.8 pg·mL–1) and the detection of acute-phase HIV in clinical human plasma samples in under 20 min. This provides a versatile absorbance-based and rapid LFIA with sensitivity capable of significantly reducing the HIV acute phase detection window. This diagnostic may be readily adapted for detection of other biomolecules as an ultrasensitive screening tool for infectious and noncommunicable diseases and can be capitalized upon in PoC settings for early disease detection. PMID:29215864

  12. Flow patterns and bathymetric signatures on the delta front of a prograding river delta

    NASA Astrophysics Data System (ADS)

    Shaw, J.; Mohrig, D. C.; Wagner, R. W.

    2016-02-01

    The transition of water between laterally confined channels and the unchannelized delta front controls the growth pattern of river deltas, but is difficult to measure on field-scale deltas. We quantify flow patterns, bathymetry and bathymetric evolution for the subaqueous delta front on the Wax Lake Delta (WLD), a rapidly prograding delta in coastal Louisiana. The flow direction field, mapped using streaklines composed of biogenic slicks on the water surface, shows that a significant portion of flow ( 59%) departs subaqueous channels laterally over the subaqueous margins of the channel seaward of the shoreline. Synoptic datasets of bathymetry and flow direction allow spatial changes in flow velocity to be quantified. Most lateral flow divergence and deceleration occurs within 3-8 channel widths outboard of subaqueous channel margins, rather than downstream of channel tips. In interdistributary bays, deposit elevation decreases with a basinward slope of 2.4 x 10-4 with distance from a channel margin along any flow path. Flow patterns and this slope produce constructional features called interdistributary troughs - topographic lows in the center of interdistributary bays. These data show that flow patterns and bathymetry on the delta front are coupled both at the transition from channelized to unchannelized flow and in the depositional regions outside the distributary network.

  13. Overview of online two-dimensional liquid chromatography based on cell membrane chromatography for screening target components from traditional Chinese medicines.

    PubMed

    Muhammad, Saqib; Han, Shengli; Xie, Xiaoyu; Wang, Sicen; Aziz, Muhammad Majid

    2017-01-01

    Cell membrane chromatography is a simple, specific, and time-saving technique for studying drug-receptor interactions, screening of active components from complex mixtures, and quality control of traditional Chinese medicines. However, the short column life, low sensitivity, low column efficiency (so cannot resolve satisfactorily mixture of compounds), low peak capacity, and inefficient in structure identification were bottleneck in its application. Combinations of cell membrane chromatography with multidimensional chromatography such as two-dimensional liquid chromatography and high sensitivity detectors like mass have significantly reduced many of the above-mentioned shortcomings. This paper provides an overview of the current advances in online two-dimensional-based cell membrane chromatography for screening target components from traditional Chinese medicines with particular emphasis on the instrumentation, preparation of cell membrane stationary phase, advantages, and disadvantages compared to alternative approaches. The last section of the review summarizes the applications of the online two-dimensional high-performance liquid chromatography based cell membrane chromatography reported since its emergence to date (2010-June 2016). © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Differentiating organic and conventional sage by chromatographic and mass spectrometry flow-injection fingerprints

    USDA-ARS?s Scientific Manuscript database

    High performance liquid chromatography (UPLC) and flow injection electrospray ionization with ion trap mass spectrometry (FIMS) fingerprints combined with the principal component analysis (PCA) were examined for their potential in differentiating commercial organic and conventional sage samples. The...

  15. Hydrodynamic interaction of two deformable drops in confined shear flow.

    PubMed

    Chen, Yongping; Wang, Chengyao

    2014-09-01

    We investigate hydrodynamic interaction between two neutrally buoyant circular drops in a confined shear flow based on a computational fluid dynamics simulation using the volume-of-fluid method. The rheological behaviors of interactive drops and the flow regimes are explored with a focus on elucidation of underlying physical mechanisms. We find that two types of drop behaviors during interaction occur, including passing-over motion and reversing motion, which are governed by the competition between the drag of passing flow and the entrainment of reversing flow in matrix fluid. With the increasing confinement, the drop behavior transits from the passing-over motion to reversing motion, because the entrainment of the reversing-flow matrix fluid turns to play the dominant role. The drag of the ambient passing flow is increased by enlarging the initial lateral separation due to the departure of the drop from the reversing flow in matrix fluid, resulting in the emergence of passing-over motion. In particular, a corresponding phase diagram is plotted to quantitatively illustrate the dependence of drop morphologies during interaction on confinement and initial lateral separation.

  16. Gas chromatography in space

    NASA Technical Reports Server (NTRS)

    Akapo, S. O.; Dimandja, J. M.; Kojiro, D. R.; Valentin, J. R.; Carle, G. C.

    1999-01-01

    Gas chromatography has proven to be a very useful analytical technique for in situ analysis of extraterrestrial environments as demonstrated by its successful operation on spacecraft missions to Mars and Venus. The technique is also one of the six scientific instruments aboard the Huygens probe to explore Titan's atmosphere and surface. A review of gas chromatography in previous space missions and some recent developments in the current environment of fiscal constraints and payload size limitations are presented.

  17. The use of laterally vectored thrust to counter thrust asymmetry in a tactical jet aircraft

    NASA Technical Reports Server (NTRS)

    1983-01-01

    A nonlinear, six degree-of-freedom flight simulator for a twin engine tactical jet was built on a hybrid computer to investigate lateral vectoring of the remaining thrust component for the case of a single engine failure at low dynamic pressures. Aircraft control was provided by an automatic controller rather than a pilot, and thrust vector control was provided by an open-loop controller that deflected a vane (located on the periphery of each exhaust jet and normally streamlined for noninterference with the flow). Lateral thrust vectoring decreased peak values of lateral control deflections, eliminated the requirement for steady-state lateral aerodynamic control deflections, and decreased the amount of altitude lost for a single engine failure.

  18. Counter-current motion in counter-current chromatography.

    PubMed

    Ito, Yoichiro

    2014-12-12

    After the CCC2012 meeting, I have received an e-mail regarding the terminology of "Countercurrent Chromatography". It stated that the term "Countercurrent" is a misnomer, because its stationary phase is motionless in the column and that the method should be renamed as liquid-liquid separations or centrifugal separations. However, it was found that these names are already used for various other techniques as found via Google search. The term "Countercurrent Chromatography" was originally made after two preparative methods of Countercurrent distribution and liquid Chromatography, both having no countercurrent motion in the column. However, it is surprising to find that this F1 hybrid method "Countercurrent Chromatography" can clearly exhibit countercurrent motion within the separation column in both hydrodynamic and hydrostatic equilibrium systems. This justifies that "Countercurrent Chromatography" is a proper term for this chromatographic method. Published by Elsevier B.V.

  19. Recovery of viral RNA and infectious foot-and-mouth disease virus from positive lateral-flow devices.

    PubMed

    Fowler, Veronica L; Bankowski, Bartlomiej M; Armson, Bryony; Di Nardo, Antonello; Valdazo-Gonzalez, Begoña; Reid, Scott M; Barnett, Paul V; Wadsworth, Jemma; Ferris, Nigel P; Mioulet, Valérie; King, Donald P

    2014-01-01

    Foot-and-mouth disease Virus (FMDV) is an economically important, highly contagious picornavirus that affects both wild and domesticated cloven hooved animals. In developing countries, the effective laboratory diagnosis of foot-and-mouth disease (FMD) is often hindered by inadequate sample preservation due to difficulties in the transportation and storage of clinical material. These factors can compromise the ability to detect and characterise FMD virus in countries where the disease is endemic. Furthermore, the high cost of sending infectious virus material and the biosecurity risk it presents emphasises the need for a thermo-stable, non-infectious mode of transporting diagnostic samples. This paper investigates the potential of using FMDV lateral-flow devices (LFDs) for dry transportation of clinical samples for subsequent nucleic acid amplification, sequencing and recovery of infectious virus by electroporation. FMDV positive samples (epithelial suspensions and cell culture isolates) representing four FMDV serotypes were applied to antigen LFDs: after which it was possible to recover viral RNA that could be detected using real-time RT-PCR. Using this nucleic acid, it was also possible to recover VP1 sequences and also successfully utilise protocols for amplification of complete FMD virus genomes. It was not possible to recover infectious FMDV directly from the LFDs, however following electroporation into BHK-21 cells and subsequent cell passage, infectious virus could be recovered. Therefore, these results support the use of the antigen LFD for the dry, non-hazardous transportation of samples from FMD endemic countries to international reference laboratories.

  20. A Point-of-Need infrared mediated PCR platform with compatible lateral flow strip for HPV detection.

    PubMed

    Liu, Wenjia; Zhang, Mingfang; Liu, Xiaoyan; Sharma, Alok; Ding, Xianting

    2017-10-15

    With the increasing need of monitoring the epidemiology of serious infectious diseases, food hygiene, food additives and pesticide residues, it is urgent to develop portable, easy-to-use, inexpensive and rapid molecular diagnostic tools. Herein, we demonstrate a prototype of IR mediated Conducting Oil and CarbOn Nanotube circUlaTing PCR (IR-COCONUT PCR) platform for nucleic acid amplification. The presented platform offers a new solution for miniaturized PCR instruments with non-contact heaters by using conducting oil and carbon nanotube as a medium in IR mediated PCR. This novel platform offers accurate and flexible control of temperature through the integration of PID (proportional-integral-derivative) algorithms to manipulate the duty cycle of the voltage signals of IR LED and a peristaltic pump. The ramping rate of the introduced platform in current study is 1.5°C/s for heating speed and -2.0°C/s for cooling speed. This platform fulfills 30 thermal cycles within 50min which is a match to the conventional bench-top PCR thermo cyclers. For demonstration purpose, human papillomavirus (HPV) patient cervical swab specimens were examined. Downstream lateral flow strip (LFS) was also developed to quantity the PCR products from the IR-COCONUT PCR device within 25min. This PCR platform together with the compatible LFS shows great potential for in-field and Point-of-Need (PoN) testing of genetic or contagious diseases. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Micellar liquid chromatography

    NASA Astrophysics Data System (ADS)

    Basova, Elena M.; Ivanov, Vadim M.; Shpigun, Oleg A.

    1999-12-01

    Background and possibilities of practical applications of micellar liquid chromatography (MLC) are considered. Various retention models in MLC, the effects of the nature and concentration of surfactants and organic modifiers, pH, temperature and ionic strength on the MLC efficiency and selectivity are discussed. The advantages and limitations of MLC are demonstrated. The performance of MLC is critically evaluated in relationship to the reversed-phase HPLC and ion-pair chromatography. The potential of application of MLC for the analysis of pharmaceuticals including that in biological fluids and separation of inorganic anions, transition metal cations, metal chelates and heteropoly compounds is described. The bibliography includes 146 references.

  2. Coupling nanoliter high-performance liquid chromatography to inductively coupled plasma mass spectrometry for arsenic speciation.

    PubMed

    Cheng, Heyong; Shen, Lihuan; Liu, Jinhua; Xu, Zigang; Wang, Yuanchao

    2018-04-01

    Nanoliter high-performance liquid chromatography shows low consumption of solvents and samples, offering one of the best choices for arsenic speciation in precious samples in combination with inuctively coupled plasma mass spectrometry. A systematic investigation on coupling nanoliter high-performance liquid chromatography to inductively coupled plasma mass spectrometry from instrument design to injected sample volume and mobile phase was performed in this study. Nanoflow mobile phase was delivered by flow splitting using a conventional high-pressure pump with reuse of mobile phase waste. Dead volume was minimized to 60 nL for the sheathless interface based on the previously developed nanonebulizer. Capillary columns for nanoliter high-performance liquid chromatography were found to be sensitive to sample loading volume. An apparent difference was also found between the mobile phases for nanoliter and conventional high-performance liquid chromatography. Baseline separation of arsenite, arsenate, monomethylarsenic, and dimethylarsenic was achieved within 11 min on a 15 cm C 18 capillary column and within 12 min on a 25 cm strong anion exchange column. Detection limits of 0.9-1.8 μg/L were obtained with precisions variable in the range of 1.6-4.2%. A good agreement between determined and certified values of a certified reference material of human urine (GBW 09115) validated its accuracy along with good recoveries (87-102%). © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Study of flow behavior in all-vanadium redox flow battery using spatially resolved voltage distribution

    NASA Astrophysics Data System (ADS)

    Bhattarai, Arjun; Wai, Nyunt; Schweiss, Rüdiger; Whitehead, Adam; Scherer, Günther G.; Ghimire, Purna C.; Nguyen, Tam D.; Hng, Huey Hoon

    2017-08-01

    Uniform flow distribution through the porous electrodes in a flow battery cell is very important for reducing Ohmic and mass transport polarization. A segmented cell approach can be used to obtain in-situ information on flow behaviour, through the local voltage or current mapping. Lateral flow of current within the thick felts in the flow battery can hamper the interpretation of the data. In this study, a new method of segmenting a conventional flow cell is introduced, which for the first time, splits up both the porous felt as well as the current collector. This dual segmentation results in higher resolution and distinct separation of voltages between flow inlet to outlet. To study the flow behavior for an undivided felt, monitoring the OCV is found to be a reliable method, instead of voltage or current mapping during charging and discharging. Our approach to segmentation is simple and applicable to any size of the cell.

  4. Chromatography: Are We Getting It Right?

    ERIC Educational Resources Information Center

    Maitland, Pamela D.; Maitland, David P.

    2002-01-01

    Explains the basics of chromatography which is used to demonstrate the separation of plant photosynthetic pigments. Reports the results of an evaluative study that explored textbook errors in explaining how chromatography works. (Contains 13 references.) (Author/YDS)

  5. Lateral asymmetry of the Hoffmann reflex: relation to cortical laterality.

    PubMed Central

    Goode, D J; Glenn, S; Manning, A A; Middleton, J F

    1980-01-01

    Lateral asymmetry of the Hoffmann reflex (H-reflex) recovery curve was found in seven subjects with no personal or family history of neurological or psychiatric disorder. Differences between recovery curves from the right and left leg were larger than differences in the same leg on two successive test days. In a group of 27 psychiatric inpatients, lateral asymmetry of the later portion of the recovery curve was correlated with cortical laterality, as measured by selective identification of differing verbal stimuli presented simultaneously to both ears (DL) and to total laterality scores, a sum of visual half-field, DL, and motor laterality scores. Asymmetry of the recovery curve is related in part to cortical laterality, possibly through selective activation of cortical motor centres on the preferred side. PMID:7420106

  6. Impact of lateral boundary conditions on regional analyses

    NASA Astrophysics Data System (ADS)

    Chikhar, Kamel; Gauthier, Pierre

    2017-04-01

    Regional and global climate models are usually validated by comparison to derived observations or reanalyses. Using a model in data assimilation results in a direct comparison to observations to produce its own analyses that may reveal systematic errors. In this study, regional analyses over North America are produced based on the fifth-generation Canadian Regional Climate Model (CRCM5) combined with the variational data assimilation system of the Meteorological Service of Canada (MSC). CRCM5 is driven at its boundaries by global analyses from ERA-interim or produced with the global configuration of the CRCM5. Assimilation cycles for the months of January and July 2011 revealed systematic errors in winter through large values in the mean analysis increments. This bias is attributed to the coupling of the lateral boundary conditions of the regional model with the driving data particularly over the northern boundary where a rapidly changing large scale circulation created significant cross-boundary flows. Increasing the time frequency of the lateral driving and applying a large-scale spectral nudging improved significantly the circulation through the lateral boundaries which translated in a much better agreement with observations.

  7. Optimizing separations in online comprehensive two‐dimensional liquid chromatography

    PubMed Central

    Gargano, Andrea F.G.; Schoenmakers, Peter J.

    2017-01-01

    Abstract Online comprehensive two‐dimensional liquid chromatography has become an attractive option for the analysis of complex nonvolatile samples found in various fields (e.g. environmental studies, food, life, and polymer sciences). Two‐dimensional liquid chromatography complements the highly popular hyphenated systems that combine liquid chromatography with mass spectrometry. Two‐dimensional liquid chromatography is also applied to the analysis of samples that are not compatible with mass spectrometry (e.g. high‐molecular‐weight polymers), providing important information on the distribution of the sample components along chemical dimensions (molecular weight, charge, lipophilicity, stereochemistry, etc.). Also, in comparison with conventional one‐dimensional liquid chromatography, two‐dimensional liquid chromatography provides a greater separation power (peak capacity). Because of the additional selectivity and higher peak capacity, the combination of two‐dimensional liquid chromatography with mass spectrometry allows for simpler mixtures of compounds to be introduced in the ion source at any given time, improving quantitative analysis by reducing matrix effects. In this review, we summarize the rationale and principles of two‐dimensional liquid chromatography experiments, describe advantages and disadvantages of combining different selectivities and discuss strategies to improve the quality of two‐dimensional liquid chromatography separations. PMID:29027363

  8. Simulating Water Flow in Variably Saturated Soils - Exploring the Advantage of Three-dimensional Models

    NASA Astrophysics Data System (ADS)

    Hopp, L.; Ivanov, V. Y.

    2010-12-01

    There is still a debate in rainfall-runoff modeling over the advantage of using three-dimensional models based on partial differential equations describing variably saturated flow vs. models with simpler infiltration and flow routing algorithms. Fully explicit 3D models are computationally demanding but allow the representation of spatially complex domains, heterogeneous soils, conditions of ponded infiltration, and solute transport, among others. Models with simpler infiltration and flow routing algorithms provide faster run times and are likely to be more versatile in the treatment of extreme conditions such as soil drying but suffer from underlying assumptions and ad-hoc parameterizations. In this numerical study, we explore the question of whether these two model strategies are competing approaches or if they complement each other. As a 3D physics-based model we use HYDRUS-3D, a finite element model that numerically solves the Richards equation for variably-saturated water flow. As an example of a simpler model, we use tRIBS+VEGGIE that solves the 1D Richards equation for vertical flow and applies Dupuit-Forchheimer approximation for saturated lateral exchange and gravity-driven flow for unsaturated lateral exchange. The flow can be routed using either the D-8 (steepest descent) or D-infinity flow routing algorithms. We study lateral subsurface stormflow and moisture dynamics at the hillslope-scale, using a zero-order basin topography, as a function of storm size, antecedent moisture conditions and slope angle. The domain and soil characteristics are representative of a forested hillslope with conductive soils in a humid environment, where the major runoff generating process is lateral subsurface stormflow. We compare spatially integrated lateral subsurface flow at the downslope boundary as well as spatial patterns of soil moisture. We illustrate situations where both model approaches perform equally well and identify conditions under which the application of a

  9. Integration of electrochemistry with ultra-performance liquid chromatography/mass spectrometry.

    PubMed

    Cai, Yi; Zheng, Qiuling; Liu, Yong; Helmy, Roy; Loo, Joseph A; Chen, Hao

    2015-01-01

    This study presents the development of ultra-performance liquid chromatography (UPLC) mass spectrometry (MS) combined with electrochemistry (EC) for the first time and its application for the structural analysis of proteins/peptides that contain disulfide bonds. In our approach, a protein/peptide mixture sample undergoes a fast UPLC separation and subsequent electrochemical reduction in an electrochemical flow cell followed by online MS and tandem mass spectrometry (MS/MS) analyses. The electrochemical cell is coupled to the mass spectrometer using our recently developed desorption electrospray ionization (DESI) interface. Using this UPLC/EC/DESI-MS method, peptides that contain disulfide bonds can be differentiated from those without disulfide bonds, as the former are electroactive and reducible. MS/MS analysis of the disulfide-reduced peptide ions provides increased information on the sequence and disulfide-linkage pattern. In a reactive DESI- MS detection experiment in which a supercharging reagent was used to dope the DESI spray solvent, increased charging was obtained for the UPLC-separated proteins. Strikingly, upon online electrolytic reduction, supercharged proteins (e.g., α-lactalbumin) showed even higher charging, which will be useful in top- down protein structure MS analysis as increased charges are known to promote protein ion dissociation. Also, the separation speed and sensitivity are enhanced by approximately 1(~)2 orders of magnitude by using UPLC for the liquid chromatography (LC)/EC/MS platform, in comparison to the previously used high- performance liquid chromatography (HPLC). This UPLC/EC/DESI-MS method combines the power of fast UPLC separation, fast electrochemical conversion, and online MS structural analysis for a potentially valuable tool for proteomics research and bioanalysis.

  10. Continuous-flow liquid microjunction surface sampling probe connected on-line with high-performance liquid chromatography/mass spectrometry for spatially resolved analysis of small molecules and proteins.

    PubMed

    Van Berkel, Gary J; Kertesz, Vilmos

    2013-06-30

    A continuous-flow liquid microjunction surface sampling probe extracts soluble material from surfaces for direct ionization and detection by mass spectrometry. Demonstrated here is the on-line coupling of such a probe with high-performance liquid chromatography/mass spectrometry (HPLC/MS) enabling extraction, separation and detection of small molecules and proteins from surfaces in a spatially resolved (~0.5 mm diameter spots) manner. A continuous-flow liquid microjunction surface sampling probe was connected to a six-port, two-position valve for extract collection and injection to an HPLC column. A QTRAP® 5500 hybrid triple quadrupole linear ion trap equipped with a Turbo V™ ion source operated in positive electrospray ionization (ESI) mode was used for all experiments. The system operation was tested with the extraction, separation and detection of propranolol and associated metabolites from drug dosed tissues, caffeine from a coffee bean, cocaine from paper currency, and proteins from dried sheep blood spots on paper. Confirmed in the tissue were the parent drug and two different hydroxypropranolol glucuronides. The mass spectrometric response for these compounds from different locations in the liver showed an increase with increasing extraction time (5, 20 and 40 s). For on-line separation and detection/identification of extracted proteins from dried sheep blood spots, two major protein peaks dominated the chromatogram and could be correlated with the expected masses for the hemoglobin α and β chains. Spatially resolved sampling, separation, and detection of small molecules and proteins from surfaces can be accomplished using a continuous-flow liquid microjunction surface sampling probe coupled on-line with HPLC/MS detection. Published in 2013. This article is a U.S. Government work and is in the public domain in the USA.

  11. POLYCYCLIC AROMATIC HYDROCARBON FORMATION IN OPPOSED FLOW DIFFUSION FLAMES OF ETHANE. (R825412)

    EPA Science Inventory

    Abstract

    The effect of fuel-side carbon density on the levels of polycyclic aromatic hydrocarbon (PAH) formation in atmospheric pressure, opposed flow, ethane diffusion flames has been studied using heated micro-probe sampling and gas chromatography/mass spectrometry (...

  12. Fluorochemical Mass Flows in a Municipal Wastewater Treatment Facility

    PubMed Central

    Schultz, Melissa M.; Higgins, Christopher P.; Huset, Carin A.; Luthy, Richard G.; Barofsky, Douglas F.; Field, Jennifer A.

    2008-01-01

    Fluorochemicals have widespread applications and are released into municipal wastewater treatment plants via domestic wastewater. A field study was conducted at a full-scale municipal wastewater treatment plant to determine the mass flows of selected fluorochemicals. Flow-proportional, 24-h samples of raw influent, primary effluent, trickling filter effluent, secondary effluent, and final effluent and grab samples of primary, thickened, activated, and anaerobically-digested sludge were collected over ten days and analyzed by liquid chromatography electrospray-ionization tandem mass spectrometry. Significant decreases in the mass flows of perfluorohexane sulfonate and perfluorodecanoate occurred during trickling filtration and primary clarification, while activated sludge treatment decreased the mass flow of perfluorohexanoate. Mass flows of the 6:2 fluorotelomer sulfonate and perfluorooctanoate were unchanged as a result of wastewater treatment, which indicates that conventional wastewater treatment is not effective for removal of these compounds. A net increase in the mass flows for perfluorooctane and perfluorodecane sulfonates occurred from trickling filtration and activated sludge treatment. Mass flows for perfluoroalkylsulfonamides and perfluorononanoate also increased during activated sludge treatment and are attributed to degradation of precursor molecules. PMID:17180988

  13. Analysis of trace dicyandiamide in stream water using solid phase extraction and liquid chromatography UV spectrometry.

    PubMed

    Qiu, Huidong; Sun, Dongdi; Gunatilake, Sameera R; She, Jinyan; Mlsna, Todd E

    2015-09-01

    An improved method for trace level quantification of dicyandiamide in stream water has been developed. This method includes sample pretreatment using solid phase extraction. The extraction procedure (including loading, washing, and eluting) used a flow rate of 1.0mL/min, and dicyandiamide was eluted with 20mL of a methanol/acetonitrile mixture (V/V=2:3), followed by pre-concentration using nitrogen evaporation and analysis with high performance liquid chromatography-ultraviolet spectroscopy (HPLC-UV). Sample extraction was carried out using a Waters Sep-Pak AC-2 Cartridge (with activated carbon). Separation was achieved on a ZIC(®)-Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) (50mm×2.1mm, 3.5μm) chromatography column and quantification was accomplished based on UV absorbance. A reliable linear relationship was obtained for the calibration curve using standard solutions (R(2)>0.999). Recoveries for dicyandiamide ranged from 84.6% to 96.8%, and the relative standard deviations (RSDs, n=3) were below 6.1% with a detection limit of 5.0ng/mL for stream water samples. Copyright © 2015. Published by Elsevier B.V.

  14. Development of a cerebrospinal fluid lateral reservoir model in rhesus monkeys (Macaca mulatta).

    PubMed

    Lester McCully, Cynthia M; Bacher, John; MacAllister, Rhonda P; Steffen-Smith, Emilie A; Saleem, Kadharbatcha; Thomas, Marvin L; Cruz, Rafael; Warren, Katherine E

    2015-02-01

    Rapid, serial, and humane collection of cerebrospinal fluid (CSF) in nonhuman primates (NHP) is an essential element of numerous research studies and is currently accomplished via two different models. The CSF reservoir model (FR) combines a catheter in the 4th ventricle with a flexible silastic reservoir to permit circulating CSF flow. The CSF lateral port model (LP) consists of a lateral ventricular catheter and an IV port that provides static access to CSF and volume restrictions on sample collection. The FR model is associated with an intensive, prolonged recovery and frequent postsurgical hydrocephalus and nonpatency, whereas the LP model is associated with an easier recovery. To maximize the advantages of both systems, we developed the CSF lateral reservoir model (LR), which combines the beneficial features of the 2 previous models but avoids their limitations by using a reservoir for circulating CSF flow combined with catheter placement in the lateral ventricle. Nine adult male rhesus monkeys were utilized in this study. Pre-surgical MRI was performed to determine the coordinates of the lateral ventricle and location of choroid plexus (CP). The coordinates were determined to avoid the CP and major blood vessels. The predetermined coordinates were 100% accurate, according to MRI validation. The LR system functioned successfully in 67% of cases for 221 d, and 44% remain functional at 426 to 510 d postoperatively. Compared with established models, our LR model markedly reduced postoperative complications and recovery time. Development of the LR model was successful in rhesus macaques and is a useful alternative to the FR and LP methods of CSF collection from nonhuman primates.

  15. Thin layer chromatography residue applicator sampler

    DOEpatents

    Nunes, Peter J [Danville, CA; Kelly, Fredrick R [Modesto, CA; Haas, Jeffrey S [San Ramon, CA; Andresen, Brian D [Livermore, CA

    2007-07-24

    A thin layer chromatograph residue applicator sampler. The residue applicator sampler provides for rapid analysis of samples containing high explosives, chemical warfare, and other analyses of interest under field conditions. This satisfied the need for a field-deployable, small, hand-held, all-in-one device for efficient sampling, sample dissolution, and sample application to an analytical technique. The residue applicator sampler includes a sampling sponge that is resistant to most chemicals and is fastened via a plastic handle in a hermetically sealed tube containing a known amount of solvent. Upon use, the wetted sponge is removed from the sealed tube and used as a swiping device across an environmental sample. The sponge is then replaced in the hermetically sealed tube where the sample remains contained and dissolved in the solvent. A small pipette tip is removably contained in the hermetically sealed tube. The sponge is removed and placed into the pipette tip where a squeezing-out of the dissolved sample from the sponge into the pipette tip results in a droplet captured in a vial for later instrumental analysis, or applied directly to a thin layer chromatography plate for immediate analysis.

  16. A novel stationary phase derivatized from hydrophilic gigaporous polystyrene-based microspheres for high-speed protein chromatography.

    PubMed

    Qu, Jian-Bo; Wan, Xing-Zhong; Zhai, Yan-Qin; Zhou, Wei-Qing; Su, Zhi-Guo; Ma, Guang-Hui

    2009-09-11

    Using agarose coated gigaporous polystyrene microspheres as a base support, a novel anion exchanger (DEAE-AP) has been developed after functionalization with diethylaminoethyl chloride. The gigaporous structure, static adsorption behavior, and chromatographic properties of DEAE-AP medium were characterized and compared with those of commercially available resin DEAE Sepharose Fast Flow (DEAE-FF). The results implied that there existed some through pores in DEAE-AP microspheres, which effectively reduced resistance to stagnant mobile phase mass transfer by inducing convective flow of mobile phase in the gigapores of medium. As a consequence, the column packed with DEAE-AP exhibited low column backpressure, high column efficiency, high dynamic binding capacity and high protein resolution at high flow velocity up to 2600cm/h. In conclusion, all the results suggested that the gigaporous absorbent is promising for high-speed protein chromatography.

  17. Differentiating organic from conventional peppermints using chromatographic and flow-injection mass spectrometric (FIMS) fingerprints

    USDA-ARS?s Scientific Manuscript database

    High performance liquid chromatography (HPLC) and flow-injection mass spectrometric (FIMS) fingerprinting techniques were tested for their potential in differentiating organic and conventional peppermint samples. Ten organic and ten conventional peppermint samples were examined using HPLC-UV and FI...

  18. Method using gas chromatography to determine the molar flow balance for proton exchange membrane fuel cells exposed to impurities

    NASA Astrophysics Data System (ADS)

    Bender, G.; Angelo, M.; Bethune, K.; Dorn, S.; Thampan, T.; Rocheleau, R.

    An understanding of the potentially serious performance degradation effects that trace level contaminants can cause in proton exchange membrane fuel cells (PEMFCs) is crucial for the successful deployment of PEMFC for commercial applications. An experimental and analytic methodology is described that employs gas chromatography (GC) to accurately determine the concentration of impurity species in the fuel and oxidant streams of a PEMFC. In this paper we further show that the accurate determination of the contaminant concentrations at the anode and cathode inlets and outlets provides a means to quantify reactions of contaminants within the cell and to identify diffusive mass transport across the membrane. High data accuracy down to sub-ppm contaminant levels is required and was achieved by addressing several challenges pertaining to experimental setup and data analysis which are both discussed in detail. The application of the methodology is demonstrated using carbon monoxide and toluene which were injected into the cell at concentrations between 1 and 10 ppm and 20 and 60 ppm, respectively. Both impurities were observed to react in the fuel cell: carbon monoxide to carbon dioxide, and toluene to methylcyclohexane. For both contaminants closure of the molar flow balances to within 3% was achieved even at the low contaminant concentrations. This allowed the extent of both reactions at the applied operating conditions to be quantified. The presented methodology is shown to be a valuable tool for investigating the effects and reactions of trace contaminants in fuel cells and for providing critical insights into the mechanisms responsible for the associated performance degradation.

  19. Magnetic Lateral Flow Strip for the Detection of Cocaine in Urine by Naked Eyes and Smart Phone Camera.

    PubMed

    Wu, Jing; Dong, Mingling; Zhang, Cheng; Wang, Yu; Xie, Mengxia; Chen, Yiping

    2017-06-05

    Magnetic lateral flow strip (MLFS) based on magnetic bead (MB) and smart phone camera has been developed for quantitative detection of cocaine (CC) in urine samples. CC and CC-bovine serum albumin (CC-BSA) could competitively react with MB-antibody (MB-Ab) of CC on the surface of test line of MLFS. The color of MB-Ab conjugate on the test line relates to the concentration of target in the competition immunoassay format, which can be used as a visual signal. Furthermore, the color density of the MB-Ab conjugate can be transferred into digital signal (gray value) by a smart phone, which can be used as a quantitative signal. The linear detection range for CC is 5-500 ng/mL and the relative standard deviations are under 10%. The visual limit of detection was 5 ng/mL and the whole analysis time was within 10 min. The MLFS has been successfully employed for the detection of CC in urine samples without sample pre-treatment and the result is also agreed to that of enzyme-linked immunosorbent assay (ELISA). With the popularization of smart phone cameras, the MLFS has large potential in the detection of drug residues in virtue of its stability, speediness, and low-cost.

  20. Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography

    PubMed Central

    Dutta, Amit K.; Tran, Travis; Napadensky, Boris; Teella, Achyuta; Brookhart, Gary; Ropp, Philip A.; Zhang, Ada W.; Tustian, Andrew D.; Zydney, Andrew L.; Shinkazh, Oleg

    2015-01-01

    Recent studies using simple model systems have demonstrated that Continuous Countercurrent Tangential Chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an “after binder” to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (~0.67 g/L) and one with high titer (~6.9 g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to that obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products. PMID:25747172

  1. Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography.

    PubMed

    Dutta, Amit K; Tran, Travis; Napadensky, Boris; Teella, Achyuta; Brookhart, Gary; Ropp, Philip A; Zhang, Ada W; Tustian, Andrew D; Zydney, Andrew L; Shinkazh, Oleg

    2015-11-10

    Recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an "after binder" to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (∼ 0.67 g/L) and one with high titer (∼ 6.9 g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to those obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Purification of lignans from Fructus Arctii using off-line two-dimensional supercritical fluid chromatography/reversed-phase liquid chromatography.

    PubMed

    Yang, Bichao; Xin, Huaxia; Wang, Feier; Cai, Jianfeng; Liu, Yanfang; Fu, Qing; Jin, Yu; Liang, Xinmiao

    2017-08-01

    As a common traditional Chinese medicine, Fructus Arctii has important clinical medical values. Its main components are lignans, which are difficult to separate and analyze because of the complex composition, similar chemical structures, and close properties. In this study, an off-line two-dimensional supercritical fluid chromatography/reversed-phase liquid chromatography method, as well as an effective sample pretreatment method based on hydrophilic interaction chromatography material, was developed to enrich the minor lignan fractions and obtain high-purity compounds. In total, 12 high-purity compounds were isolated from Fructus Arctii. Their structures were identified by using high-resolution mass spectrometry and nuclear magnetic resonance spectroscopy, which showed that all were lignans and that most of them were isomers. The results demonstrated the effective off-line two-dimensional supercritical fluid chromatography/reversed-phase liquid chromatography method for the purification of lignans from Fructus Arctii. The separation protocol established here will be beneficial for the separation of complex samples from other kinds of natural products. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Cerebral Lateralization and General Intelligence: Gender Differences in a Transcranial Doppler Study

    ERIC Educational Resources Information Center

    Njemanze, P.C.

    2005-01-01

    The present study evaluated cerebral lateralization during Raven's progressive matrices (RPM) paradigm in female and male subjects. Bilateral simultaneous transcranial Doppler (TCD) ultrasound was used to measure mean blood flow velocities (MBFV) in the right and left middle cerebral arteries (MCAs) in 24 (15 females and 9 males) right-handed…

  4. Loop-Mediated Isothermal Amplification-Lateral-Flow Dipstick (LAMP-LFD) to detect Toxoplasma gondii oocyst in ready-to-eat salad.

    PubMed

    Lalle, Marco; Possenti, Alessia; Dubey, Jitender P; Pozio, Edoardo

    2018-04-01

    The apicomplexan parasite Toxoplasma gondii is the causative agent of toxoplasmosis, a foodborne zoonosis with a global distribution and estimated to cause up to 20% of the total foodborne disease burden in Europe. Association between T. gondii infection and the consumption of unwashed raw fruits and vegetables contaminated with oocysts has been reported and the increasing habit to eat pre-washed ready-to-eat salads poses a new potential risk for consumers. It is therefore important to trace the occurrence of potential contamination with this parasite to guarantee the safety of ready-to-eat vegetables. Detection of T. gondii in vegetables by molecular techniques has been achieved but low sensitivity (PCR) or expensive equipments (qPCR) limit routine applicability. Here, we describe the development and validation of a sensitive and robust method relying on a LAMP assay, targeting the 529 bp locus, to detect T. gondii oocysts down to 25 oocysts/50 g in ready-to-eat baby lettuce. The LAMP has been also adapted for a faster visualization of the result by a lateral flow dipstick chromatographic detection method. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Optimizing separations in online comprehensive two-dimensional liquid chromatography.

    PubMed

    Pirok, Bob W J; Gargano, Andrea F G; Schoenmakers, Peter J

    2018-01-01

    Online comprehensive two-dimensional liquid chromatography has become an attractive option for the analysis of complex nonvolatile samples found in various fields (e.g. environmental studies, food, life, and polymer sciences). Two-dimensional liquid chromatography complements the highly popular hyphenated systems that combine liquid chromatography with mass spectrometry. Two-dimensional liquid chromatography is also applied to the analysis of samples that are not compatible with mass spectrometry (e.g. high-molecular-weight polymers), providing important information on the distribution of the sample components along chemical dimensions (molecular weight, charge, lipophilicity, stereochemistry, etc.). Also, in comparison with conventional one-dimensional liquid chromatography, two-dimensional liquid chromatography provides a greater separation power (peak capacity). Because of the additional selectivity and higher peak capacity, the combination of two-dimensional liquid chromatography with mass spectrometry allows for simpler mixtures of compounds to be introduced in the ion source at any given time, improving quantitative analysis by reducing matrix effects. In this review, we summarize the rationale and principles of two-dimensional liquid chromatography experiments, describe advantages and disadvantages of combining different selectivities and discuss strategies to improve the quality of two-dimensional liquid chromatography separations. © 2017 The Authors. Journal of Separation Science published by WILEY-VCH Verlag GmbH & Co. KGaA.

  6. Application of ionic liquids in liquid chromatography and electrodriven separation.

    PubMed

    Huang, Yi; Yao, Shun; Song, Hang

    2013-08-01

    Ionic liquids (ILs) are salts in the liquid state at ambient temperature, which are nonvolatile, nonflammable with high thermal stability and dissolve easily for a wide range of inorganic and organic materials. As a kind of potential green solvent, they show high efficiency and selectivity in the field of separation research, especially in instrumental analysis. Thus far, ILs have been successfully applied by many related researchers in high-performance liquid chromatography and capillary electrophoresis as chromatographic stationary phases, mobile phase additives or electroosmotic flow modifiers. This paper provides a detailed review of these applications in the study of natural products, foods, drugs and other fine chemicals. Furthermore, the prospects of ILs in liquid chromatographic and electrodriven techniques are discussed.

  7. Demonstration and properties of a planar heterojunction bipolar transistor with lateral current flow

    NASA Astrophysics Data System (ADS)

    Thornton, Robert L.; Mosby, William J.; Chung, Harlan F.

    1989-10-01

    The authors present fabrication techniques and device performance for a novel transistor structure, the lateral heterojunction bipolar transistor. The lateral heterojunctions are formed by impurity-induced disordering of a GaAs base layer sandwiched between two AlGaAs layers. These transistor structures exhibit current gains of 14 for base widths of 0.74 micron. Transistor action in this device occurs parallel to the surface of the device structure. The active base region of the structure is completely submerged, resulting in a reduction of surface recombination as a mechanism for gain reduction in the device. Impurity-induced disordering is used to widen the bandgap of the alloy in the emitter and collector, resulting in an improvement of the emitter injection efficiency. Since the device is based entirely on a surface diffusion process, the device is completely planar and has no steps involving etching of the III-V alloy material. These advantages lead this device to be considered as a candidate for optoelectronic integration applications. The transistor device functions as a buried heterostructure laser, with a threshold current as low as 6 mA for a 1.4-micron stripe.

  8. Simultaneous enantioselective determination of triadimefon and its metabolite triadimenol in edible vegetable oil by gel permeation chromatography and ultraperformance convergence chromatography/tandem mass spectrometry.

    PubMed

    Yao, Zhoulin; Li, Xiaoge; Miao, Yelong; Lin, Mei; Xu, Mingfei; Wang, Qiang; Zhang, Hu

    2015-11-01

    A novel, sensitive, and efficient enantioselective method for the determination of triadimefon and its metabolite triadimenol in edible vegetable oil, was developed by gel permeation chromatography and ultraperformance convergence chromatography/tandem triple quadrupole mass spectrometry. After the vegetable oil samples were prepared using gel permeation chromatography, the eluent was collected, evaporated, and dried with nitrogen gas. The residue was redissolved by adding methanol up to a final volume of 1 mL. The analytes of six enantiomers were analyzed on Chiralpak IA-3 column (150 × 4.6 mm) using compressed liquid CO2-mixed 14 % co-solvents, comprising methanol/acetonitrile/isopropanol = 20/20/60 (v/v/v) in the mobile phase at 30 °C, and the total separation time was less than 4 min at a flow rate of 2 mL/min. Quantification was achieved using matrix-matched standard calibration curves. The overall mean recoveries for six enantiomers from vegetable oil were 90.1-97.3 %, with relative standard deviations of 0.8-5.4 % intra-day and 2.3-5.0 % inter-day at 0.5, 5, and 50 μg/kg levels. The limits of quantification were 0.5 μg/kg for all enantiomers based on five replicate extractions at the lowest fortified level in vegetable oil. Moreover, the absolute configuration of six enantiomers had been determined based on comparisons of the vibrational circular dichroism experimental spectra with the theoretical curve obtained by density functional theory calculations. Application of the proposed method to the 40 authentic vegetable oil samples from local markets suggests its potential use in enantioselective determination of triadimefon and triadimenol enantiomers. Graphical Abstract Chemical structures and UPC(2)-MS/MS separation chromatograms of triadimefon and triadimenol.

  9. Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis.

    PubMed

    Zhang, Qian; Yang, Feng-Qing; Ge, Liya; Hu, Yuan-Jia; Xia, Zhi-Ning

    2017-01-01

    Hydrophilic interaction liquid chromatography, an alternative liquid chromatography mode, is of particular interest in separating hydrophilic and polar ionic compounds. Compared with traditional liquid chromatography techniques, hydrophilic interaction liquid chromatography offers specific advantages mainly including: (1) relatively green and water-soluble mobile phase composition, which enhances the solubility of hydrophilic and polar ionic compounds; (2) no need for ion-pairing reagents and high content of organic solvent, which benefits mass spectrometry detection; (3) high orthogonality to reverse-phase liquid chromatography, well adapted to two-dimensional liquid chromatography for complicated samples. Therefore, hydrophilic interaction liquid chromatography has been rapidly developed in many areas over the past decades. This review summarizes the recent progress (from 2012 to July 2016) of hydrophilic interaction liquid chromatography in pharmaceutical analysis, with the focus on detecting chemical drugs in various matrices, charactering active compounds of natural products and assessing biotherapeutics through typical structure unit. Moreover, the retention mechanism and behavior of analytes in hydrophilic interaction liquid chromatography as well as some novel hydrophilic interaction liquid chromatography columns used for pharmaceutical analysis are also described. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. There and Back Again: Development and Regeneration of the Zebrafish Lateral Line System

    PubMed Central

    Thomas, Eric D.; Cruz, Ivan A.; Hailey, Dale W.; Raible, David W.

    2014-01-01

    The zebrafish lateral line is a sensory system used to detect changes in water flow. It is comprised of clusters of mechanosensory hair cells called neuromasts. The lateral line is initially established by a migratory group of cells, called a primordium, that deposits neuromasts at stereotyped locations along the surface of the fish. Wnt, FGF, and Notch signaling are all important regulators of various aspects of lateral line development, from primordium migration to hair cell specification. As zebrafish age, the organization of the lateral line becomes more complex in order to accommodate the fish’s increased size. This expansion is regulated by many of the same factors involved in the initial development. Furthermore, unlike mammalian hair cells, lateral line hair cells have the capacity to regenerate after damage. New hair cells arise from the proliferation and differentiation of surrounding support cells, and the molecular and cellular pathways regulating this are beginning to be elucidated. All in all, the zebrafish lateral line has proven to be an excellent model in which to study a diverse array of processes, including collective cell migration, cell polarity, cell fate, and regeneration. PMID:25330982

  11. A novel superporous agarose medium for high-speed protein chromatography.

    PubMed

    Shi, Qing-Hong; Zhou, Xin; Sun, Yan

    2005-12-05

    A novel superporous agarose (SA) bead characterized by the presence of wide pores has been fabricated by water-in-oil emulsification using solid granules of calcium carbonate as porogenic agent. After cross-linking, the solid granules were removed by dissolving them in hydrochloric acid. Then, the gel was modified with diethylaminoethyl groups to create an anion exchanger, SA-DEAE, for protein adsorption. A homogeneous agarose (HA) bead was also produced and modified with DEAE for comparison. It was found that the porosity of SA-DEAE was about 6% larger than that of HA-DEAE. Moreover, both optical micrographs and confocal laser scanning microscopy (CLSM) of the ion exchangers with adsorbed fluorescein isothiocyanate (FITC) labeled IgG revealed the superporous structure of the SA medium. In addition, the SA-DEAE column had lower backpressure than the HA-DEAE column, confirming the convective flow of mobile phase through the wide pores. Due to the presence of the wide pores, more channels were available for protein transport and, furthermore, more diffusive pores in the agarose network were accessible for the protein approach from different directions. This led to 40% higher protein capacity and two times higher effective pore diffusivity in the SA-DEAE than in HA-DEAE. Moreover, an increase of the efficiency of the SA-DEAE column until a flow rate of 5 cm/min and the independency of the column efficiency at flow rates from 5 to 17.8 cm/min was found, indicating that intraparticle mass transfer was intensified by convective flow at elevated flow rates. Therefore, the chromatographic resolution of IgG and BSA was little affected up to a flow rate of 17.8 cm/min. The results indicate that the SA medium is favorable for high-speed protein chromatography. (c) 2005 Wiley Periodicals, Inc.

  12. High-mobility capacitively-induced two-dimensional electrons in a lateral superlattice potential

    DOE PAGES

    Lu, Tzu -Ming; Laroche, Dominique; Huang, S. -H.; ...

    2016-01-01

    In the presence of a lateral periodic potential modulation, two-dimensional electrons may exhibit interesting phenomena, such as a graphene-like energy-momentum dispersion, Bloch oscillations, or the Hofstadter butterfly band structure. To create a sufficiently strong potential modulation using conventional semiconductor heterostructures, aggressive device processing is often required, unfortunately resulting in strong disorder that masks the sought-after effects. Here, we report a novel fabrication process flow for imposing a strong lateral potential modulation onto a capacitively induced two-dimensional electron system, while preserving the host material quality. Using this process flow, the electron density in a patterned Si/SiGe heterostructure can be tuned overmore » a wide range, from 4.4 × 10 10 cm –2 to 1.8 × 10 11 cm –2, with a peak mobility of 6.4 × 10 5 cm 2/V·s. The wide density tunability and high electron mobility allow us to observe sequential emergence of commensurability oscillations as the density, the mobility, and in turn the mean free path, increase. Magnetic-field-periodic quantum oscillations associated with various closed orbits also emerge sequentially with increasing density. We show that, from the density dependence of the quantum oscillations, one can directly extract the steepness of the imposed superlattice potential. Lastly, this result is then compared to a conventional lateral superlattice model potential.« less

  13. Topographically driven crustal flow and its implication to the development of pinned oroclines

    NASA Technical Reports Server (NTRS)

    Hsui, Albert T.; Wilkerson, M. Scott; Marshak, Stephen

    1990-01-01

    Pinned oroclines, a type of curved orogen which results from lateral pinning of a growing fold-thrust belt, tend to resemble parabolic Newtonian curvature modified by different degrees of flattening at the flow front. It is proposed that such curves can be generated by Newtonian crustal flow driven by topographic variations. In this model, regional topographic differences create a regional flow which produces a parabolic flow front on interaction with lateral bounding obstacles. Local topographic variations modify the parabolic curves and yield more flat-crested, non-Newtonian-type curvatures. A finite-difference thin-skin tectonic simulation demonstrates that both Newtonian and non-Newtonian curved orogens can be produced within a Newtonian crust.

  14. Supercritical fluid chromatography versus high performance liquid chromatography for enantiomeric and diastereoisomeric separations on coated polysaccharides-based stationary phases: Application to dihydropyridone derivatives.

    PubMed

    Hoguet, Vanessa; Charton, Julie; Hecquet, Paul-Emile; Lakhmi, Chahinaze; Lipka, Emmanuelle

    2018-05-11

    For analytical applications, SFC has always remained in the shadow of LC. Analytical enantioseparation of eight dihydropyridone derivatives, was run in both High Performance Liquid Chromatography and Supercritical Fluid Chromatography. Four polysaccharide based chiral stationary phases namely amylose and cellulose tris(3, 5-dimethylphenylcarbamate), amylose tris((S)-α-phenylethylcarbamate) and cellulose tris(4-methylbenzoate) with four mobile phases consisted of either n-hexane/ethanol or propan-2-ol (80:20 v:v) or carbon dioxide/ethanol or propan-2-ol (80:20 v:v) mixtures were investigated under same operatory conditions (temperature and flow-rate). The elution strength, enantioselectivity and resolution were compared in the two methodologies. For these compounds, for most of the conditions, HPLC afforded shorter retention times and a higher resolution than SFC. HPLC appears particularly suitable for the separation of the compounds bearing two chiral centers. For instance compound 7 was baseline resolved on OD-H CSP under n-Hex/EtOH 80/20, with resolution values equal to 2.98, 1.55, 4.52, between the four stereoisomers in less than 17 min, whereas in SFC, this latter is not fully separated in 23 min under similar eluting conditions. After analytical screenings, the best conditions were transposed to semi-preparative scale. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Preparative enantiomer separation of dichlorprop with a cinchona-derived chiral selector employing centrifugal partition chromatography and high-performance liquid chromatography: a comparative study.

    PubMed

    Gavioli, Elena; Maier, Norbert M; Minguillón, Cristina; Lindner, Wolfgang

    2004-10-01

    A countercurrent chromatography protocol for support-free preparative enantiomer separation of the herbicidal agent 2-(2,4-dichlorphenoxy)propionic acid (dichlorprop) was developed utilizing a purposefully designed, highly enantioselective chiral stationary-phase additive (CSPA) derived from bis-1,4-(dihydroquinidinyl)phthalazine. Guided by liquid-liquid extraction experiments, a solvent system consisting of 10 mM CSPA in methyl tert-butyl ether and 100 mM sodium phosphate buffer (pH 8.0) was identified as a suitable stationary/mobile-phase combination. This solvent system provided an ideal compromise among stationary-phase retention, enantioselectivity, and well-balanced analyte distribution behavior. Using a commercial centrifugal partition chromatography instrument, complete enantiomer separations of up to 366 mg of racemic dichlorprop could be achieved, corresponding to a sample load being equivalent to the molar amount of CSPA employed. Comparison of the preparative performance characteristics of the CPC protocol with that of a HPLC separation using a silica-supported bis-1,4-(dihydroquinidinyl)phthalazine chiral stationary phase CSP revealed comparable loading capacities for both techniques but a significantly lower solvent consumption for CPC. With respect to productivity, HPLC was found to be superior, mainly due to inherent flow rate restrictions of the CPC instrument. Given that further progress in instrumental design and engineering of dedicated, highly enantioselective CSPAs can be achieved, CPC may offer a viable alternative to CSP-based HPLC for preparative-scale enantiomer separation.

  16. Flow structures and sandbar dynamics in a canyon river during a controlled flood, Colorado River, Arizona

    USGS Publications Warehouse

    Wright, S.A.; Kaplinski, M.

    2011-01-01

    In canyon rivers, debris fan constrictions create rapids and downstream pools characterized by secondary flow structures that are closely linked to channel morphology. In this paper we describe detailed measurements of the three-dimensional flow structure and sandbar dynamics of two pools along the Colorado River in the Grand Canyon during a controlled flood release from Glen Canyon Dam. Results indicate that the pools are characterized by large lateral recirculation zones (eddies) resulting from flow separation downstream from the channel constrictions, as well as helical flow structures in the main channel and eddy. The lateral recirculation zones are low-velocity areas conducive to fine sediment deposition, particularly in the vicinity of the separation and reattachment points and are thus the dominant flow structures controlling sandbar dynamics. The helical flow structures also affect morphology but appear secondary in importance to the lateral eddies. During the controlled flood, sandbars in the separation and reattachment zones at both sites tended to build gradually during the rising limb and peak flow. Deposition in shallow water on the sandbars was accompanied by erosion in deeper water along the sandbar slope at the interface with the main channel. Erosion occurred via rapid mass failures as well as by gradual boundary shear stress driven processes. The flow structures and morphologic links at our study sites are similar to those identified in other river environments, in particular sharply curved meanders and channel confluences where the coexistence of lateral recirculation and helical flows has been documented. Copyright 2011 by the American Geophysical Union.

  17. Isolation and purification of six iridoid glycosides from gardenia jasminoides fruit by medium-pressure liquid chromatography combined with macroporous resin chromatography.

    PubMed

    Wang, Yun; Liu, Hui; Shen, Lifeng; Yao, Lan; Ma, Yinlian; Yu, Dingrong; Chen, Jianhong; Li, Puling; Chen, Ying; Zhang, Cun

    2015-12-01

    Gardeniae fructus is one of the most frequently used herbs in traditional Chinese medicine. In the present study, a process for the enrichment of six iridoid glycosides from Gardeniae fructus was developed using medium-pressure liquid chromatography combined with macroporous resin and reversed-phase chromatography. The purities of different fractions from Gardeniae fructus were assessed using quantitative high-performance liquid chromatography. After fractionation using HPD-100 column chromatography, a 30% ethanol fraction was selected based on high-performance liquid chromatography and liquid chromatography with mass spectrometry qualitative analysis to separate and purify. Based on the orientation analysis results, six compounds-deacetyl asperulosidic acid methyl ester, gardenoside, ixoroside, scandoside methyl ester, genipin-1-O-β-d-gentiobioside, and geniposide-were successfully isolated and purified in three to four combined steps from Gardeniae fructus. The purities of these compounds were found by high-performance liquid chromatography analysis to be 97.9, 98.1, 95.5, 96.3, 97.1, and 98.7%, respectively. Moreover, their structures were elucidated by NMR spectroscopy and liquid chromatography with tandem mass spectrometry. The separation process was highly efficient, rapid, and accurate, making it a potential approach for the large-scale production of iridoids in the laboratory and providing several marker compounds for quality control. This procedure may be meaningful for the purification of other natural products used in traditional Chinese medicine. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. The sensory basis of rheotaxis in turbulent flow

    NASA Astrophysics Data System (ADS)

    Elder, John P.

    Rheotaxis is a robust, multisensory behavior with many potential benefits for fish and other aquatic animals, yet the influence of different fluvial conditions on rheotactic performance and its sensory basis is still poorly understood. Here, we examine the role that vision and the lateral line play in the rheotactic behavior of a stream-dwelling species (Mexican tetra, Astyanax mexicanus) under both rectilinear and turbulent flow conditions. Turbulence enhanced overall rheotactic strength and lowered the flow speed at which rheotaxis was initiated; this effect did not depend on the availability of either visual or lateral line information. Compared to fish without access to visual information, fish with access to visual information exhibited increased levels of positional stability and as a result, increased levels of rheotactic accuracy. No disruption in rheotactic performance was found when the lateral line was disabled, suggesting that this sensory system is not necessary for either rheotaxis or turbulence detection under the conditions of this study.

  19. Quasi 3D modeling of water flow in vadose zone and groundwater

    USDA-ARS?s Scientific Manuscript database

    The complexity of subsurface flow systems calls for a variety of concepts leading to the multiplicity of simplified flow models. One habitual simplification is based on the assumption that lateral flow and transport in unsaturated zone are not significant unless the capillary fringe is involved. In ...

  20. The multi-mode modulator: A versatile fluidic device for two-dimensional gas chromatography.

    PubMed

    Seeley, John V; Schimmel, Nicolaas E; Seeley, Stacy K

    2018-02-09

    A fluidic device called the multi-mode modulator (MMM) has been developed for use as a comprehensive two-dimensional gas chromatography (GC x GC) modulator. The MMM can be employed in a wide range of capacities including as a traditional heart-cutting device, a low duty cycle GC x GC modulator, and a full transfer GC x GC modulator. The MMM is capable of producing narrow component pulses (widths <50ms) while operating at flows compatible with high resolution chromatography. The sample path of modulated components is confined to the interior of a joining capillary. The joining capillary dimensions and the position of the columns within the joining capillary can be optimized for the selected modulation mode. Furthermore, the joining capillary can be replaced easily and inexpensively if it becomes fouled due to sample matrix components or column bleed. The principles of operation of the MMM are described and its efficacy is demonstrated as a heart-cutting device and as a GC x GC modulator. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Ion Chromatography.

    ERIC Educational Resources Information Center

    Mulik, James D.; Sawicki, Eugene

    1979-01-01

    Accurate for the analysis of ions in solution, this form of analysis enables the analyst to directly assay many compounds that previously were difficult or impossible to analyze. The method is a combination of the methodologies of ion exchange, liquid chromatography, and conductimetric determination with eluant suppression. (Author/RE)

  2. Performance and limits of liquid chromatography isotope ratio mass spectrometry system for halogenated compounds

    NASA Astrophysics Data System (ADS)

    Gilevska, Tetyana; Gehre, Matthias; Richnow, Hans

    2014-05-01

    Compound Specific Isotope Analysis (CSIA) has been an important step for the assessment of the origin and fate of compounds in environmental science.[1] Biologically or pharmaceutically important compounds often are not amenable for gas chromatographic separation because of high polarity and lacking volatility, thermostability. In 2004 liquid chromatography isotope ratio mass spectrometry (LC-IRMS) became commercially available. LC-IRMS system intent a quantitative conversion of analytes separation into CO2 via wet oxidation with sodium persulfate in the presence of phosphoric acid while analytes are still dissolved in the aqueous liquid phase.[2] The aim of this study is to analyze the oxidation capacity of the interface of the LC-IRMS system and determine which parameters could improve oxidation of compounds which are resistant to persulfate oxidation. Oxidation capacity of the liquid chromatography isotope ratio mass spectrometry system was tested with halogenated acetic acid and a set of aromatic compounds with different substitutes. Acetic acid (AA) was taken as a model compound for complete oxidation and compared to the oxidation of other analytes on a molar basis. Correct values were obtained for di- and mono chlorinated and fluorinated and also for tribrominated acetic acid and for all studied aromatic compounds. Incomplete oxidation for trichloroacetic (TCAA) and trifluoroacetic (TFAA) acid was revealed with lower recovery compared to acetic acid and isotope fractionation leading to depleted carbon isotope composition compared to values obtained with an elementary analyzer connected to an isotope mass spectrometer Several optimization steps were tried in order to improve the oxidation of TCAA and TFAA: (i) increasing the concentration of the oxidizing agent, (ii) variation of flow rate of the oxidizing and acid solution, (iii) variation of flow rate of liquid chromatography pump (iv) addition of a catalyzer. These modifications lead to longer reaction time

  3. Negative chromatography purification of hepatitis B virus-like particles using poly(oligo(ethylene glycol) methacrylate) grafted cationic adsorbent.

    PubMed

    Lee, Micky Fu Xiang; Chan, Eng Seng; Tan, Wen Siang; Tam, Kam Chiu; Tey, Beng Ti

    2015-10-09

    Poly(oligo(ethylene glycol) methacrylate) (POEGMA), an inert polymer was grafted onto an anion exchange adsorbent for the exclusion of relatively larger hepatitis B virus-like particles (HB-VLPs) from the anion exchange ligand (Q) and at the same time this process allowed the selective adsorption of smaller size Escherichia coli host cell proteins (HCPs). The chain lengths of the POEGMA grafted were modulated by varying the amount of monomers used in the polymer grafting. The purification factor and yield of the HB-VLPs obtained from the flow-through of negative chromatography were 2.3 and 66.0±3.1%, respectively, when shorter chain length of POEGMA (SQ) was grafted. Adsorbent grafted with longer chain of POEGMA (LQ) excluded some HCPs that are larger in size together with the HB-VLPs, reducing the purity of the recovered HB-VLPs. Further heat-treatment of the flow-through pool from SQ followed by centrifugation increased the purity of heat stable HB-VLPs to 87.5±1.1%. Heat-treatment of the flow through sample resulted in thermal denaturation and aggregation of HCPs, while the heat stable HB-VLPs still remained intact as observed under a transmission electron microscope. The performance of the negative chromatography together with heat treatment in the purification of HB-VLPs is far better than the reported bind-and-elute techniques. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Perfusion chromatography separation of the tomato fruit-specific pectin methylesterase from a semipurified commercial enzyme preparation.

    PubMed

    Savary, B J

    2001-08-01

    A rapid and simple method was developed, using perfusion chromatography media, to separate the fruit-specific pectin methylesterase (PME) isoform from the depolymerizing enzyme polygalacturonase (PG) and other contaminating pectinases present in a commercial tomato enzyme preparation. Pectinase activities were adsorbed onto a Poros HS (a strong cation exchanger) column in 20 M HEPES buffer at pH 7.5. The fruit-specific PME was eluted from the column with 80 mM NaCl, followed by a step to 300 mM NaCl to elute PG activity. Rechromatography of the PME activity peak with a linear gradient further resolved two PME isoenzymes and removed residual traces of PG activity. The PG activity peak was further treated with lectin affinity chromatography to provide purified PG enzyme, which was separated from a salt-dependent PME (tentatively identified as a "ubiquitous-type" isoform), and a pectin acetylesterase. The later enzyme has not been reported previously in tomato. This method provides monocomponent enzymes that will be useful for studying enzyme mechanisms and for modifying pectin structure and functional properties.

  5. Improving washing strategies of human mesenchymal stem cells using negative mode expanded bed chromatography.

    PubMed

    Cunha, Bárbara; Silva, Ricardo J S; Aguiar, Tiago; Serra, Margarida; Daicic, John; Maloisel, Jean-Luc; Clachan, John; Åkerblom, Anna; Carrondo, Manuel J T; Peixoto, Cristina; Alves, Paula M

    2016-01-15

    The use of human mesenchymal stem cells (hMSC) in clinical applications has been increasing over the last decade. However, to be applied in a clinical setting hMSC need to comply with specific requirements in terms of identity, potency and purity. This study reports the improvement of established tangential flow filtration (TFF)-based washing strategies, further increasing hMSC purity, using negative mode expanded bed adsorption (EBA) chromatography with a new multimodal prototype matrix based on core-shell bead technology. The matrix was characterized and a stable, expanded bed could be obtained using standard equipment adapted from what is used for conventional packed bed chromatography processes. The effect of different expansion rates on cell recovery yield and protein removal capacity was assessed. The best trade-off between cell recovery (89%) and protein clearance (67%) was achieved using an intermediate expansion bed rate (1.4). Furthermore, we also showed that EBA chromatography can be efficiently integrated on the already established process for the downstream processing (DSP) of hMSC, where it improved the washing efficiency more than 10-fold, recovering approximately 70% of cells after global processing. This strategy showed not to impact cell viability (>95%), neither hMSC's characteristics in terms of morphology, immunophenotype, proliferation, adhesion capacity and multipotent differentiation potential. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Simultaneous determination of ascorbic acid and caffeine in commercial soft drinks using reversed-phase ultraperformance liquid chromatography.

    PubMed

    Turak, Fatma; Güzel, Remziye; Dinç, Erdal

    2017-04-01

    A new reversed-phase ultraperformance liquid chromatography method with a photodiode array detector was developed for the quantification of ascorbic acid (AA) and caffeine (CAF) in 11 different commercial drinks consisting of one energy drink and 10 ice tea drinks. Separation of the analyzed AA and CAF with an internal standard, caffeic acid, was performed on a Waters BEH C 18 column (100 mm × 2.1 mm, 1.7 μm i.d.), using a mobile phase consisting of acetonitrile and 0.2M H 3 PO 4 (11:89, v/v) with a flow rate of 0.25 mL/min and an injection volume of 1.0 μL. Calibration graphs for AA and CAF were computed from the peak area ratio of AA/internal standard and CAF/internal standard detected at 244.0 nm and 273.6 nm, respectively. The developed reversed-phase ultraperformance liquid chromatography method was validated by analyzing standard addition samples. The proposed reversed-phase ultraperformance liquid chromatography method gave us successful results for the quantitative analysis of commercial drinks containing AA and CAF substances. Copyright © 2016. Published by Elsevier B.V.

  7. Study of the flow mixing in a novel ARID raceway for algae production

    DOE PAGES

    Xu, Ben; Li, Peiwen; Waller, P.

    2014-07-31

    A novel flow field for algae raceways has been proposed, which is fundamentally different from traditional paddlewheel-driven raceways. To reduce freezing and heat loss in the raceway during cold time, the water is drained to a deep storage canal. The ground bed of the new raceway has a low slope so that water, lifted by propeller pump, can flow down in laterally-laid serpentine channels, relying on gravitational force. The flow rate of water is controlled so that it can overflow the lateral channel walls and mix with the main flow in the next lower channel, which thus creates a bettermore » mixing. In order to optimize the design parameters of the new flow field, methods including flow visualization, local point velocity measurement, and CFD analysis were employed to investigate the flow mixing features. Different combinations of channel geometries and water velocities were evaluated. An optimized flow field design and details of flow mixing are presented. The study offers an innovative design for large scale algae growth raceways which is of significance to the algae and biofuel industry.« less

  8. Study of the flow mixing in a novel ARID raceway for algae production

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Xu, Ben; Li, Peiwen; Waller, P.

    A novel flow field for algae raceways has been proposed, which is fundamentally different from traditional paddlewheel-driven raceways. To reduce freezing and heat loss in the raceway during cold time, the water is drained to a deep storage canal. The ground bed of the new raceway has a low slope so that water, lifted by propeller pump, can flow down in laterally-laid serpentine channels, relying on gravitational force. The flow rate of water is controlled so that it can overflow the lateral channel walls and mix with the main flow in the next lower channel, which thus creates a bettermore » mixing. In order to optimize the design parameters of the new flow field, methods including flow visualization, local point velocity measurement, and CFD analysis were employed to investigate the flow mixing features. Different combinations of channel geometries and water velocities were evaluated. An optimized flow field design and details of flow mixing are presented. The study offers an innovative design for large scale algae growth raceways which is of significance to the algae and biofuel industry.« less

  9. Applicability of a diffusion model to lateral transport in the terrestrial and lunar exospheres.

    NASA Technical Reports Server (NTRS)

    Hodges, R. R., Jr.

    1972-01-01

    Kinetic theory is used to determine a series expansion of the vertical flux of particles in an exosphere in terms of time and space derivatives of particle concentration, exobase velocity, and temperature. For sufficiently large scale variations of these parameters in time and space, the series can be truncated to a form that is similar to a diffusion equation. Owing to this analogy, it is possible to unite the mathematical description of molecular diffusion, which governs thermospheric flow, and the corresponding exospheric equation by using effective transport coefficients which change smoothly with altitude through the transition from thermosphere to exosphere. A new definition of the exobase for lateral flow emerges from the analogy of exospheric and thermospheric diffusion, as the altitude where the horizontal mean free path length equals the mean horizontal extent of ballistic trajectories of the transported gas, as opposed to the scale height of the dominant gas which determines the exobase for escape. It is shown that the approximation of exospheric lateral flow as a diffusion process is applicable to global scale problems concerning terrestrial helium and heavier gases, and lunar gases heavier than helium.

  10. High sensitive gold-nanoparticle based lateral flow Immunodevice for Cd2+ detection in drinking waters.

    PubMed

    López Marzo, Adaris M; Pons, Josefina; Blake, Diane A; Merkoçi, Arben

    2013-09-15

    In this work for first time a lateral flow immunosensor device (LFID) for Cd(2+) determination in drinking and tap waters using the Cd-EDTA-BSA-AuNP conjugate as signal producer tool is introduced. The principle of working is based on competitive reaction between the Cd-EDTA-BSA-AuNP conjugate deposited on the conjugation pad strip and the Cd-EDTA complex formed in the analysis sample for the same binding sites of the 2A81G5 monoclonal antibody, specific to Cd-EDTA but not Cd(2+) free, which is immobilized onto the test line. The device has a large response range within 0.4-2000ppb, being the linear response between 0.4 and 10ppb. The quantification and detection limits of 0.4 and 0.1ppb, respectively, represent the lowest ones reported so far for paper based metal sensors. The obtained detection limit is 50 times lower than the maximum contamination level required for drinking water. Here we also show a new option for increasing the sensibility in the LFDs with competitive format, through the decreasing in concentrations of the Cd-EDTA-BSA-AuNP conjugate deposited in the conjugation strip and the mAbs deposited in the test and control zones until to reach optimized concentrations. It is an important result take into account that the increase in sensibility is one of the challenges in the field of LFD sensors, where are focused many of the ongoing researches. In addition, a specificity study of the device for several metal interferences, where potential metal interferences are masked with the use of the EDTA and OVA optimized concentrations, is presented too. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Development and Validation of a Lateral Flow Immunoassay for Rapid Detection of NDM-Producing Enterobacteriaceae

    PubMed Central

    Boutal, Hervé; Naas, Thierry; Devilliers, Karine; Oueslati, Saoussen; Bernabeu, Sandrine; Simon, Stéphanie

    2017-01-01

    ABSTRACT The global spread of carbapenemase-producing Enterobacteriaceae (CPE) that are often resistant to most, if not all, classes of antibiotics is a major public health concern. The NDM-1 carbapenemase is among the most worrisome carbapenemases given its rapid worldwide spread. We have developed and evaluated a lateral flow immunoassay (LFIA) (called the NDM LFIA) for the rapid and reliable detection of NDM-like carbapenemase-producing Enterobacteriaceae from culture colonies. We evaluated the NDM LFIA using 175 reference enterobacterial isolates with characterized β-lactamase gene content and 74 nonduplicate consecutive carbapenem-resistant clinical isolates referred for expertise to the French National Reference Center (NRC) for Antibiotic Resistance during a 1-week period (in June 2016). The reference collection included 55 non-carbapenemase producers and 120 carbapenemase producers, including 27 NDM producers. All 27 NDM-like carbapenemase producers of the reference collection were correctly detected in less than 15 min by the NDM LFIA, including 22 strains producing NDM-1, 2 producing NDM-4, 1 producing NDM-5, 1 producing NDM-7, and 1 producing NDM-9. All non-NDM-1 producers gave a negative result with the NDM LFIA. No cross-reaction was observed with carbapenemases (VIM, IMP, NDM, KPC, and OXA-48-like), extended-spectrum β-lactamases (ESBLs) (TEM, SHV, and CTX-M), AmpCs (CMY-2, DHA-2, and ACC-1), and oxacillinases (OXA-1, -2, -9, and -10). Similarly, among the 74 referred nonduplicate consecutive clinical isolates, all 7 NDM-like producers were identified. Overall, the sensitivity and specificity of the assay were 100% for NDM-like carbapenemase detection with strains cultured on agar. The NDM LFIA was efficient, rapid, and easy to implement in the routine workflow of a clinical microbiology laboratory for the confirmation of NDM-like carbapenemase-producing Enterobacteriaceae. PMID:28404680

  12. Lipopolysaccharide Specific Immunochromatography Based Lateral Flow Assay for Serogroup Specific Diagnosis of Leptospirosis in India

    PubMed Central

    Vanithamani, Shanmugam; Shanmughapriya, Santhanam; Narayanan, Ramasamy; Raja, Veerapandian; Kanagavel, Murugesan; Sivasankari, Karikalacholan; Natarajaseenivasan, Kalimuthusamy

    2015-01-01

    Background Leptospirosis is a re-emerging infectious disease that is under-recognized due to low-sensitivity and cumbersome serological tests. MAT is the gold standard test and it is the only serogroup specific test used till date. Rapid reliable alternative serogroup specific tests are needed for surveillance studies to identify locally circulating serogroups in the study area. Methods/Principal Findings In the present investigation the serological specificity of leptospiral lipopolysaccharides (LPS) was evaluated by enzyme linked immunosorbent assay (ELISA), dot blot assay and rapid immunochromatography based lateral flow assay (ICG-LFA). Sera samples from 120 MAT positive cases, 174 cases with febrile illness other than leptospirosis, and 121 seronegative healthy controls were evaluated for the diagnostic sensitivity and specificity of the developed assays. LPS was extracted from five locally predominant circulating serogroups including: Australis (27.5%), Autumnalis (11.7%), Ballum (25.8%), Grippotyphosa (12.5%), Pomona (10%) and were used as antigens in the diagnostics to detect IgM antibodies in patients’ sera. The sensitivity observed by IgM ELISA and dot blot assay using various leptospiral LPS was >90% for homologous sera. Except for Ballum LPS, no other LPS showed cross-reactivity to heterologous sera. An attempt was made to develop LPS based ICG-LFA for rapid and sensitive serogroup specific diagnostics of leptospirosis. The developed ICG-LFA showed sensitivity in the range between 93 and 100% for homologous sera. The Wilcoxon analysis showed LPS based ICG-LFA did not differ significantly from the gold standard MAT (P>0.05). Conclusion The application of single array of LPS for serogroup specific diagnosis is first of its kind. The developed assay could potentially be evaluated and employed for as MAT alternative. PMID:26340095

  13. Two opposed lateral jets injected into swirling crossflow

    NASA Technical Reports Server (NTRS)

    Lilley, D. G.; Mcmurry, C. B.; Ong, L. H.

    1987-01-01

    Experiments have been conducted to obtain the time-mean and turbulent quantities of opposed lateral jets in a low speed, nonreacting flowfield. A jet-to-crossflow velocity ratio of R = 4 was used throughout the experiments, with swirl vane angles of 0 (swirler removed), 45 and 70 degrees used with the crossflow. Flow visualization techniques used were neutrally-buoyant helium-filled soap bubbles and multispark photography in order to obtain the gross flowfield characteristics. Measurements of time-mean and turbulent quantities were obtained utilizing a six-orientation single hot-wire technique. For the nonswirling case, the jets were found not to penetrate past the test-section centerline, in contrast to the single lateral jet with the same jet-to-crossflow velocity ratio. In the swirling cases, the crossflow remains in a narrow region near the wall of the test section. The opposed jets are swept from their vertical courses into spiral trajectories close to the confining walls. Extensive results are presented in r-x plane plots.

  14. Role of surface and subsurface lateral water flows on summer precipitation in a complex terrain region: A WRF-Hydro case-study for Southern Germany

    NASA Astrophysics Data System (ADS)

    Rummler, Thomas; Arnault, Joel; Gochis, David; Kunstmann, Harald

    2017-04-01

    Recent developments in hydrometeorological modeling aim towards more sophisticated treatment of terrestrial hydrologic processes. The standard version of the Weather Research and Forecasting (WRF) model describes terrestrial water transport as a purely vertical process. The hydrologically enhanced version of WRF, namely WRF-Hydro, does account for lateral terrestrial water flows, which allows for a more comprehensive process description of the interdependencies between water- and energy fluxes at the land-atmosphere interface. In this study, WRF and WRF-Hydro are applied to the Bavarian Alpine region in southern Germany, a complex terrain landscape in a relatively humid, mid-latitude climate. Simulation results are validated with gridded and station observation of precipitation, temperature and river discharge. Differences between WRF and WRF-Hydro results are investigated with a joint atmospheric-terrestrial water budget analysis. Changes in the partitioning in (near-) surface runoff and percolation are prominent. However, values for evapotranspiration ET feature only marginal variations, suggesting that soil moisture content is not a limiting factor of ET in this specific region. Simulated precipitation fields during isolated summertime events still show appreciable differences, while differences in large-scale, multi-day rainy periods are less substantial. These differences are mainly related to differences in the moisture in- and outflow terms of the atmospheric water budget induced by the surface and sub-surface lateral redistribution of soil moisture in WRF-Hydro.

  15. Highly sensitive and selective lateral flow immunoassay based on magnetic nanoparticles for quantitative detection of carcinoembryonic antigen.

    PubMed

    Liu, Fangming; Zhang, Honglian; Wu, Zhenhua; Dong, Haidao; Zhou, Lin; Yang, Dawei; Ge, Yuqing; Jia, Chunping; Liu, Huiying; Jin, Qinghui; Zhao, Jianlong; Zhang, Qiqing; Mao, Hongju

    2016-12-01

    Carcinoembryonic antigen (CEA) is an important biomarker in cancer diagnosis. Here, we present an efficient, selective lateral-flow immunoassay (LFIA) based on magnetic nanoparticles (MNPs) for in situ sensitive and accurate point-of-care detection of CEA. Signal amplification mechanism involved linking of detection MNPs with signal MNPs through biotin-modified single-stranded DNA (ssDNA) and streptavidin. To verify the effectiveness of this modified LFIA system, the sensitivity and specificity were evaluated. Sensitivity evaluation showed a broad detection range of 0.25-1000ng/ml for CEA protein by the modified LFIA, and the limit of detection (LOD) of the modified LFIA was 0.25ng/ml, thus producing significant increase in detection threshold compared with the traditional LFIA. The modified LFIA could selectively recognize CEA in presence of several interfering proteins. In addition, this newly developed assay was applied for quantitative detection of CEA in human serum specimens collected from 10 randomly selected patients. The modified LFIA system detected minimum 0.27ng/ml of CEA concentration in serum samples. The results were consistent with the clinical data obtained using commercial electrochemiluminescence immunoassay (ECLIA) (p<0.01). In conclusion, the MNPs based LFIA system not only demonstrated enhanced signal to noise ratio, it also detected CEA with higher sensitivity and selectivity, and thus has great potential to be commercially applied as a sensitive tumor marker filtration system. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Polymer separations by liquid interaction chromatography: principles - prospects - limitations.

    PubMed

    Radke, Wolfgang

    2014-03-28

    Most heterogeneities of polymers with respect to different structural features cannot be resolved by only size exclusion chromatography (SEC), the most frequently applied mode of polymer chromatography. Instead, methods of interaction chromatography became increasingly important. However, despite the increasing applications the principles and potential of polymer interaction chromatography are still often unknown to a large number of polymer scientists. The present review will explain the principles of the different modes of polymer chromatography. Based on selected examples it will be shown which separation techniques can be successfully applied for separations with respect to the different structural features of polymers. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Some parameters relevant to affinity chromatography on immobilized nucleotides

    PubMed Central

    Lowe, C. R.; Harvey, M. J.; Craven, D. B.; Dean, P. D. G.

    1973-01-01

    1. The suitability of cellulose and Sepharose as supports for affinity chromatography of two groups of cofactor-linked enzymes, dehydrogenases and kinases, was examined. Sepharose was found to be superior. 2. The selective capacities of the columns were measured by frontal analysis and are discussed in relation to the nucleotide contents. 3. The effect of various concentrations of enzyme and of non-specific protein on the performance of the affinity columns, and the effects of equilibration time, flow rate, sample volume and dilution of the nucleotide were examined. 4. The effect of interposing polymethylene and polyglycine extension arms between the matrix backbone and the nucleotide was investigated for several cofactor-dependent enzymes. Maximum binding was observed with an extension arm 0.8–1nm long. PMID:4354739

  18. Computational and Experimental Flow Field Analyses of Separate Flow Chevron Nozzles and Pylon Interaction

    NASA Technical Reports Server (NTRS)

    Massey, Steven J.; Thomas, Russell H.; AbdolHamid, Khaled S.; Elmiligui, Alaa A.

    2003-01-01

    A computational and experimental flow field analyses of separate flow chevron nozzles is presented. The goal of this study is to identify important flow physics and modeling issues required to provide highly accurate flow field data which will later serve as input to the Jet3D acoustic prediction code. Four configurations are considered: a baseline round nozzle with and without a pylon, and a chevron core nozzle with and without a pylon. The flow is simulated by solving the asymptotically steady, compressible, Reynolds-averaged Navier-Stokes equations using an implicit, up-wind, flux-difference splitting finite volume scheme and standard two-equation kappa-epsilon turbulence model with a linear stress representation and the addition of a eddy viscosity dependence on total temperature gradient normalized by local turbulence length scale. The current CFD results are seen to be in excellent agreement with Jet Noise Lab data and show great improvement over previous computations which did not compensate for enhanced mixing due to high temperature gradients.

  19. Soil pipe flow tracer experiments: 1. Connectivity and transport characteristics

    USDA-ARS?s Scientific Manuscript database

    Much debate has occurred in catchment hydrology regarding the connectivity of flow paths from upslope areas to catchment outlets. This study was conducted in two catchments, one with three upper branches, in a loess soil with a fragipan that fosters lateral flow and exhibits an extensive distributio...

  20. Fractionation of technical octabromodiphenyl ether by countercurrent chromatography combined with gas chromatography/mass spectrometry and offline and online (1)H nuclear magnetic resonance spectroscopy.

    PubMed

    Hammann, Simon; Conrad, Jürgen; Vetter, Walter

    2015-06-12

    Countercurrent chromatography (CCC) is a technique, which uses two immiscible liquid phases for a separation process in a long and hollow tube. The technique allows the separation of high amounts of sample (50mg to several grams) with a low consumption of solvents. In this study, we fractionated 50mg technical octabromodiphenyl ether (DE-79) and analyzed the fractions by gas chromatography with mass spectrometry (GC/MS) and proton nuclear magnetic resonance ((1)H NMR) spectroscopy. CCC separations were performed with n-hexane/acetonitrile as solvent system in tail-to-head (i.e. the upper phase is mobile) mode. Twelve CCC fractions were studied for the PBDE composition. CCC elution of PBDE congeners was dependent both on the degree of bromination and substitution pattern. Higher brominated congeners eluted faster than lower brominated congeners and isomers with vicinal hydrogen atoms eluted last. In addition to several known PBDE congeners in DE-79, we were able to unequivocally identify BDE 195 in DE-79 and we could verify the presence of BDE 184. Finally, we also established the online hyphenation of CCC with (1)H NMR. The use of deuterated solvents could be avoided by using n-hexane/acetonitrile as two-phase system. By online CCC-(1)H NMR in stop-flow mode we were able to detect eight PBDE congeners in the mixture. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. High speed Deans switch for low duty cycle comprehensive two-dimensional gas chromatography.

    PubMed

    Ghosh, Abhijit; Bates, Carly T; Seeley, Stacy K; Seeley, John V

    2013-05-24

    A new high-speed valve-based modulator has been designed and tested for use in comprehensive two-dimensional gas chromatography (GC×GC). The modulator is a Deans switch constructed from two micro-volume fittings and a solenoid valve. Modulator performance was characterized over a wide range of device settings including the magnitude of the switching flow, the gap between the tips of the primary and secondary column, the primary column flow rate, and the carrier gas identity. Under optimized conditions, the modulator was found to be capable of generating narrow pulses (<50ms) of primary effluent with a 2mL/min secondary column flow. This capability will ultimately allow the modulator to be used with GC×GC separations employing a wide range of detectors and secondary column geometries. The main disadvantage of this modulator is that it employs a low sampling duty cycle, and thus it produces separations with sensitivities that are lower than those produced with thermal modulators or differential flow modulators. The efficacy of the new high-speed Deans switch modulator was demonstrated through the GC×GC separation of a hydrocarbon standard and gasoline. Precise quantitation of individual components was possible provided the modulation ratio was kept greater than 2.0. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Purification of flavonoids from licorice using an off-line preparative two-dimensional normal-phase liquid chromatography/reversed-phase liquid chromatography method.

    PubMed

    Fan, Yunpeng; Fu, Yanhui; Fu, Qing; Cai, Jianfeng; Xin, Huaxia; Dai, Mei; Jin, Yu

    2016-07-01

    An orthogonal (71.9%) off-line preparative two-dimensional normal-phase liquid chromatography/reversed-phase liquid chromatography method coupled with effective sample pretreatment was developed for separation and purification of flavonoids from licorice. Most of the nonflavonoids were firstly removed using a self-made Click TE-Cys (60 μm) solid-phase extraction. In the first dimension, an industrial grade preparative chromatography was employed to purify the crude flavonoids. Click TE-Cys (10 μm) was selected as the stationary phase that provided an excellent separation with high reproducibility. Ethyl acetate/ethanol was selected as the mobile phase owing to their excellent solubility for flavonoids. Flavonoids co-eluted in the first dimension were selected for further purification using reversed-phase liquid chromatography. Multiple compounds could be isolated from one normal-phase fraction and some compounds with bad resolution in one-dimensional liquid chromatography could be prepared in this two-dimensional system owing to the orthogonal separation. Moreover, this two-dimensional liquid chromatography method was beneficial for the preparation of relatively trace flavonoid compounds, which were enriched in the first dimension and further purified in the second dimension. Totally, 24 flavonoid compounds with high purity were obtained. The results demonstrated that the off-line two-dimensional liquid chromatography method was effective for the preparative separation and purification of flavonoids from licorice. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Value for controlling flow of cryogenic fluid

    DOEpatents

    Knapp, Philip A.

    1996-01-01

    A valve is provided for accurately controlling the flow of cryogenic fluids such as liquid nitrogen. The valve comprises a combination of disc and needle valves affixed to a valve stem in such a manner that the disc and needle are free to rotate about the stem, but are constrained in lateral and vertical movements. This arrangement provides accurate and precise fluid flow control and positive fluid isolation.

  4. Local Modelling of Groundwater Flow Using Analytic Element Method Three-dimensional Transient Unconfined Groundwater Flow With Partially Penetrating Wells and Ellipsoidal Inhomogeneites

    NASA Astrophysics Data System (ADS)

    Jankovic, I.; Barnes, R. J.; Soule, R.

    2001-12-01

    The analytic element method is used to model local three-dimensional flow in the vicinity of partially penetrating wells. The flow domain is bounded by an impermeable horizontal base, a phreatic surface with recharge and a cylindrical lateral boundary. The analytic element solution for this problem contains (1) a fictitious source technique to satisfy the head and the discharge conditions along the phreatic surface, (2) a fictitious source technique to satisfy specified head conditions along the cylindrical boundary, (3) a method of imaging to satisfy the no-flow condition across the impermeable base, (4) the classical analytic solution for a well and (5) spheroidal harmonics to account for the influence of the inhomogeneities in hydraulic conductivity. Temporal variations of the flow system due to time-dependent recharge and pumping are represented by combining the analytic element method with a finite difference method: analytic element method is used to represent spatial changes in head and discharge, while the finite difference method represents temporal variations. The solution provides a very detailed description of local groundwater flow with an arbitrary number of wells of any orientation and an arbitrary number of ellipsoidal inhomogeneities of any size and conductivity. These inhomogeneities may be used to model local hydrogeologic features (such as gravel packs and clay lenses) that significantly influence the flow in the vicinity of partially penetrating wells. Several options for specifying head values along the lateral domain boundary are available. These options allow for inclusion of the model into steady and transient regional groundwater models. The head values along the lateral domain boundary may be specified directly (as time series). The head values along the lateral boundary may also be assigned by specifying the water-table gradient and a head value at a single point (as time series). A case study is included to demonstrate the application

  5. Nanoparticle Analysis by Online Comprehensive Two-Dimensional Liquid Chromatography combining Hydrodynamic Chromatography and Size-Exclusion Chromatography with Intermediate Sample Transformation

    PubMed Central

    2017-01-01

    Polymeric nanoparticles have become indispensable in modern society with a wide array of applications ranging from waterborne coatings to drug-carrier-delivery systems. While a large range of techniques exist to determine a multitude of properties of these particles, relating physicochemical properties of the particle to the chemical structure of the intrinsic polymers is still challenging. A novel, highly orthogonal separation system based on comprehensive two-dimensional liquid chromatography (LC × LC) has been developed. The system combines hydrodynamic chromatography (HDC) in the first-dimension to separate the particles based on their size, with ultrahigh-performance size-exclusion chromatography (SEC) in the second dimension to separate the constituting polymer molecules according to their hydrodynamic radius for each of 80 to 100 separated fractions. A chip-based mixer is incorporated to transform the sample by dissolving the separated nanoparticles from the first-dimension online in tetrahydrofuran. The polymer bands are then focused using stationary-phase-assisted modulation to enhance sensitivity, and the water from the first-dimension eluent is largely eliminated to allow interaction-free SEC. Using the developed system, the combined two-dimensional distribution of the particle-size and the molecular-size of a mixture of various polystyrene (PS) and polyacrylate (PACR) nanoparticles has been obtained within 60 min. PMID:28745485

  6. Ultrasensitive SERS Flow Detector Using Hydrodynamic Focusing

    PubMed Central

    Negri, Pierre; Jacobs, Kevin T.; Dada, Oluwatosin O.; Schultz, Zachary D.

    2013-01-01

    Label-free, chemical specific detection in flow is important for high throughput characterization of analytes in applications such as flow injection analysis, electrophoresis, and chromatography. We have developed a surface-enhanced Raman scattering (SERS) flow detector capable of ultrasensitive optical detection on the millisecond time scale. The device employs hydrodynamic focusing to improve SERS detection in a flow channel where a sheath flow confines analyte molecules eluted from a fused silica capillary over a planar SERS-active substrate. Increased analyte interactions with the SERS substrate significantly improve detection sensitivity. The performance of this flow detector was investigated using a combination of finite element simulations, fluorescence imaging, and Raman experiments. Computational fluid dynamics based on finite element analysis was used to optimize the flow conditions. The modeling indicates that a number of factors, such as the capillary dimensions and the ratio of the sheath flow to analyte flow rates, are critical for obtaining optimal results. Sample confinement resulting from the flow dynamics was confirmed using wide-field fluorescence imaging of rhodamine 6G (R6G). Raman experiments at different sheath flow rates showed increased sensitivity compared with the modeling predictions, suggesting increased adsorption. Using a 50-millisecond acquisitions, a sheath flow rate of 180 μL/min, and a sample flow rate of 5 μL/min, a linear dynamic range from nanomolar to micromolar concentrations of R6G with a LOD of 1 nM is observed. At low analyte concentrations, rapid analyte desorption is observed, enabling repeated and high-throughput SERS detection. The flow detector offers substantial advantages over conventional SERS-based assays such as minimal sample volumes and high detection efficiency. PMID:24074461

  7. Revisiting resolution in hydrodynamic countercurrent chromatography: tubing bore effect.

    PubMed

    Berthod, A; Faure, K

    2015-04-17

    A major challenge in countercurrent chromatography (CCC), the technique that works with a support-free biphasic liquid system, is to retain the liquid stationary phase inside the CCC column (Sf parameter). Two solutions are commercially available: the hydrostatic CCC columns, also called centrifugal partition chromatographs (CPC), with disks of interconnected channels and rotary seals, and the hydrodynamic CCC columns with bobbins of coiled open tube and no rotary seals. It was demonstrated that the amount of liquid stationary phase retained by a coiled tube was higher with larger bore tubing than with small bore tubes. At constant column volume, small bore tubing will be longer producing more efficiency than larger bore tube that will better retain the liquid stationary phase. Since the resolution equation in CCC is depending on both column efficiency and stationary phase retention ratio, the influence of the tubing bore should be studied. This theoretical work showed that there is an optimum tubing bore size depending on solute partition coefficient and mobile phase flow rate. The interesting result of the theoretical study is that larger tubing bores allow for dramatically reduced experiment durations for all solutes: in reversed phase CCC (polar mobile phase), hydrophobic solutes are usually highly retained. These apolar solutes can be separated by the same coil at high flow rates and reduced Sf with similar retention times as polar solutes separated at smaller flow rates and much higher Sf. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Acoustic energy exchange through flow turning

    NASA Astrophysics Data System (ADS)

    Baum, Joseph D.

    1987-01-01

    A numerical investigation of the mechanisms of acoustic energy exchange between the mean and acoustic flow fields in resonance chambers, such as rocket engines, is reported. A noniterative linearized block implicit scheme was used to solve the time-dependent compressible Navier-Stokes equations. Two test cases were investigated: acoustic wave propagation in a tube with a coexisting sheared mean flow (the refraction test) and acoustic wave propagation in a tube where the mean sheared flow was injected into the tube through its lateral boundary (the flow turning study). For flow turning, significant excitation of mean flow energy was observed at two locations: at the edge of the acoustic boundary layer and within a zone adjacent to the acoustic boundary layer extending up to 0.1 radii away from the wall. A weaker streaming effect was observed for the refraction study, and only at the edge of the acoustic boundary layer. The total dissipation for the flow turning test was twice the dissipation for refraction.

  9. Fluorescently labelled multiplex lateral flow immunoassay based on cadmium-free quantum dots.

    PubMed

    Beloglazova, Natalia V; Sobolev, Aleksander M; Tessier, Mickael D; Hens, Zeger; Goryacheva, Irina Yu; De Saeger, Sarah

    2017-03-01

    A sensitive tool for simultaneous qualitative detection of two mycotoxins based on use of non-cadmium quantum dots (QDs) is presented for the first time. QDs have proven themselves as promising fluorescent labels for biolabeling and chemical analysis. With an increasing global tendency to regulate and limit the use of hazardous elements, indium phosphide (InP) QDs are highlighted as environmentally-friendly alternatives to the highly efficient and well-studied, but potentially toxic Cd- and Pb-based QDs. Here, we developed water-soluble InP QDs-based fluorescent nanostructures. They consisted of core/shell InP/ZnS QDs enrobed in a silica shell that allowed the water solubility (QD@SiO 2 ). Then we applied the QD@SiO 2 as novel, silica shell-encapsulated fluorescent labels in immunoassays for rapid multiplexed screening. Two mycotoxins, zearalenone and deoxynivalenol, were simultaneously detected in maize and wheat, since the two QD@SiO 2 labelled conjugates emit at two different, individually detectable wavelengths. The cutoff values for the simultaneous determination were 50 and 500μgkg -1 for zearalenone and deoxynivalenol, respectively, in both maize and wheat. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was used to confirm the result. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Rapid visual detection of Mycobacterium avium subsp. paratuberculosis by recombinase polymerase amplification combined with a lateral flow dipstick

    PubMed Central

    Zhao, Guimin; Wang, Hongmei; Hou, Peili; He, Chengqiang

    2018-01-01

    Paratuberculosis (Johne's disease) is a chronic debilitating disease of domestic and wild ruminants. However, widespread point-of-care testing is infrequent due to the lack of a robust method. The isothermal recombinase polymerase amplification (RPA) technique has applied for rapid diagnosis. Herein, RPA combined with a lateral flow dipstick (LFD) assay was developed to estimate DNA from Mycobacterium avium subsp. paratuberculosis. First, analytical specificity and sensitivity of the RPA-nfo primer and probe sets were assessed. The assay successfully detected M. paratuberculosis DNA in 30 min at 39℃ with a detection limit of up to eight copies per reaction, which was equivalent to that of the real-time quantitative polymerase chain reaction (qPCR) assay. The assay was specific, as it did not amplify genomes from five other Mycobacterium spp. or five pathogenic enteric bacteria. Six hundred-twelve clinical samples (320 fecal and 292 serum) were assessed by RPA-LFD, qPCR, and enzyme-linked immunosorbent assay, respectively. The RPA-LFD assay yielded 100% sensitivity, 97.63% specificity, and 98.44% concordance rate with the qPCR results. This is the first report utilizing an RPA-LFD assay to visualize and rapidly detect M. paratuberculosis. Our results show this assay should be a useful method for the diagnosis of paratuberculosis in resource-constrained settings. PMID:29284204

  11. Rapid visual detection of Mycobacterium avium subsp. paratuberculosis by recombinase polymerase amplification combined with a lateral flow dipstick.

    PubMed

    Zhao, Guimin; Wang, Hongmei; Hou, Peili; He, Chengqiang; He, Hongbin

    2018-03-31

    Paratuberculosis (Johne's disease) is a chronic debilitating disease of domestic and wild ruminants. However, widespread point-of-care testing is infrequent due to the lack of a robust method. The isothermal recombinase polymerase amplification (RPA) technique has applied for rapid diagnosis. Herein, RPA combined with a lateral flow dipstick (LFD) assay was developed to estimate DNA from Mycobacterium avium subsp. paratuberculosis . First, analytical specificity and sensitivity of the RPA-nfo primer and probe sets were assessed. The assay successfully detected M. paratuberculosis DNA in 30 min at 39°C with a detection limit of up to eight copies per reaction, which was equivalent to that of the real-time quantitative polymerase chain reaction (qPCR) assay. The assay was specific, as it did not amplify genomes from five other Mycobacterium spp. or five pathogenic enteric bacteria. Six hundred-twelve clinical samples (320 fecal and 292 serum) were assessed by RPA-LFD, qPCR, and enzyme-linked immunosorbent assay, respectively. The RPA-LFD assay yielded 100% sensitivity, 97.63% specificity, and 98.44% concordance rate with the qPCR results. This is the first report utilizing an RPA-LFD assay to visualize and rapidly detect M. paratuberculosis . Our results show this assay should be a useful method for the diagnosis of paratuberculosis in resource-constrained settings.

  12. The Influence of Laterality of Pharyngeal Bolus Passage on Dysphagia in Hemiplegic Stroke Patients

    PubMed Central

    Kim, Min Seok; Lee, Seong Jae; Seo, Dong Hyuk; Hyun, Jung Keun; Kim, Jae Il

    2012-01-01

    Objective To investigate swallowing laterality in hemiplegic patients with stroke and recovery of dysphagia according to the laterality. Method The sample was comprised of 46 dysphagic patients with hemiplegia after their first stroke. The sample's videofluoroscopic swallowing study (VFSS) was reviewed. Swallowing laterality was determined by the anterior-posterior view of VFSS. We measured width difference of barium sulfate liquid flow in the pharyngoesophageal segment. If there was double or more the width of that from the opposite width in the pharyngoesophageal segment more than twice on three trials of swallowing, then it was judged as having laterality. Subjects were assigned to no laterality (NL), laterality that is ipsilateral to hemiplegic side (LI), and laterality that is contralateral to hemiplegic side (LC) groups. We measured the following: prevalence of aspiration, the 8-point penetration-aspiration scale, and the functional dysphagia scale of the subjects at baseline and follow up. Results Laterality was observed in 45.7% of all patients. Among them, 52.4% were in the hemiplegic direction. There was no significant difference between groups at baseline in all measurements. When we compared the changes in all measurements on follow-up study, there were no significant differences between groups. Conclusion Through this study, we found that there was no significant relation between swallowing laterality and the severity or prognosis of swallowing difficulty. More studies for swallowing laterality on stroke patients will be needed. PMID:23185735

  13. Three-dimensional inspiratory flow in a double bifurcation airway model

    NASA Astrophysics Data System (ADS)

    Jalal, Sahar; Nemes, Andras; Van de Moortele, Tristan; Schmitter, Sebastian; Coletti, Filippo

    2016-09-01

    The flow in an idealized airway model is investigated for the steady inhalation case. The geometry consists of a symmetric planar double bifurcation that reflects the anatomical proportions of the human bronchial tree, and a wide range of physiologically relevant Reynolds numbers ( Re = 100-5000) is considered. Using magnetic resonance velocimetry, we analyze the three-dimensional fields of velocity and vorticity, along with flow descriptors that characterize the longitudinal and lateral dispersion. In agreement with previous studies, the symmetry of the flow partitioning is broken even at the lower Reynolds numbers, and at the second bifurcation, the fluid favors the medial branches over the lateral ones. This trend reaches a plateau around Re = 2000, above which the turbulent inflow results in smoothed mean velocity gradients. This also reduces the streamwise momentum flux, which is a measure of the longitudinal dispersion by the mean flow. The classic Dean-type counter-rotating vortices are observed in the first-generation daughter branches as a result of the local curvature. In the granddaughter branches, however, the secondary flows are determined by the local curvature only for the lower flow regimes ( Re ≤ 250), in which case the classic Dean mechanism prevails. At higher flow regimes, the field is instead dominated by streamwise vortices extending from the daughter into the medial granddaughter branches, where they rotate in the opposite direction with respect to Dean vortices. Circulation and secondary flow intensity show a similar trend as the momentum flux, increasing with Reynolds number up to Re = 2000 and then dropping due to turbulent dissipation of vorticity. The streamwise vortices interact both with each other and with the airway walls, and for Re > 500 they can become stronger in the medial granddaughter than in the upstream daughter branches. With respect to realistic airway models, the idealized geometry produces weaker secondary flows

  14. Spiral counter-current chromatography of small molecules, peptides and proteins using the spiral tubing support rotor.

    PubMed

    Knight, Martha; Finn, Thomas M; Zehmer, John; Clayton, Adam; Pilon, Aprile

    2011-09-09

    An important advance in countercurrent chromatography (CCC) carried out in open flow-tubing coils, rotated in planetary centrifuges, is the new design to spread out the tubing in spirals. More spacing between the tubing was found to significantly increase the stationary phase retention, such that now all types of two-phase solvent systems can be used for liquid-liquid partition chromatography in the J-type planetary centrifuges. A spiral tubing support (STS) frame with circular channels was constructed by laser sintering technology into which FEP tubing was placed in 4 spiral loops per layer from the bottom to the top and a cover affixed allowing the tubing to connect to flow-tubing of the planetary centrifuge. The rotor was mounted and run in a P.C. Inc. type instrument. Examples of compounds of molecular weights ranging from <300 to approximately 15,000 were chromatographed in appropriate two-phase solvent systems to assess the capability for separation and purification. A mixture of small molecules including aspirin was completely separated in hexane-ethyl acetate-methanol-water. Synthetic peptides including a very hydrophobic peptide were each purified to a very high purity level in a sec-butanol solvent system. In the STS rotor high stationary phase retention was possible with the aqueous sec-butanol solvent system at a normal flow rate. Finally, the two-phase aqueous polyethylene glycol-potassium phosphate solvent system was applied to separate a protein from a lysate of an Escherichia coli expression system. These experiments demonstrate the versatility of spiral CCC using the STS rotor. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Helium microwave-induced plasmas for element specific detection in chromatography

    NASA Astrophysics Data System (ADS)

    Long, Gary L.; Ducatte, Gerald R.; Lancaster, Edwin D.

    1994-01-01

    This review addresses the development and use of the microwave-induced plasma (MIP) using optical emission spectrometry for the purpose of element specific detection in chromatography. The plasma source that is the focus of this article is the He-based MIP operated at atmospheric pressure. The forms of chromatography that are covered include gas chromatography, liquid chromatography, and supercritical fluid chromatography. Concepts in plasma cavity design and the chromatograph-plasma interface are discussed along with the application of these hybrid systems to analytical determinations.

  16. Gas Chromatography

    NASA Astrophysics Data System (ADS)

    Qian, Michael C.

    Gas chromatography (GC) has many applications in the analysis of food products. GC has been used for the determination of fatty acids, triglycerides, cholesterol, gases, water, alcohols, pesticides, flavor compounds, and many more. While GC has been used for other food components such as sugars, oligosaccharides, amino acids, peptides, and vitamins, these substances are more suited to analysis by high performance liquid chromatography. GC is ideally suited to the analysis of volatile substances that are thermally stable. Substances such as pesticides and flavor compounds that meet these criteria can be isolated from a food and directly injected into the GC. For compounds that are thermally unstable, too low in volatility, or yield poor chromatographic separation due to polarity, a derivatization step must be done before GC analysis. The two parts of the experiment described here include the analysis of alcohols that requires no derivatization step, and the analysis of fatty acids which requires derivatization. The experiments specify the use of capillary columns, but the first experiment includes conditions for a packed column.

  17. Measurements of water uptake of maize roots: the key function of lateral roots

    NASA Astrophysics Data System (ADS)

    Ahmed, M. A.; Zarebanadkouki, M.; Kroener, E.; Kaestner, A.; Carminati, A.

    2014-12-01

    Maize (Zea mays L.) is one of the most important crop worldwide. Despite its importance, there is limited information on the function of different root segments and root types of maize in extracting water from soils. Therefore, the aim of this study was to investigate locations of root water uptake in maize. We used neutron radiography to: 1) image the spatial distribution of maize roots in soil and 2) trace the transport of injected deuterated water (D2O) in soil and roots. Maizes were grown in aluminum containers (40×38×1 cm) filled with a sandy soil. When the plants were 16 days old, we injected D2O into selected soil regions containing primary, seminal and lateral roots. The experiments were performed during the day (transpiring plants) and night (not transpiring plants). The transport of D2O into roots was simulated using a new convection-diffusion numerical model of D2O transport into roots. By fitting the observed D2O transport we quantified the diffusional permeability and the water uptake of the different root segments. The maize root architecture consisted of a primary root, 4-5 seminal roots and many lateral roots connected to the primary and seminal roots. Laterals emerged from the proximal 15 cm of the primary and seminal roots. Water uptake occurred primarily in lateral roots. Lateral roots had the highest diffusional permeability (9.4×10-7), which was around six times higher that the diffusional permeability of the old seminal segments (1.4×10-7), and two times higher than the diffusional permeability of the young seminal segments (4.7×10-7). The radial flow of D2O into the lateral (6.7×10-5 ) was much higher than in the young seminal roots (1.1×10-12). The radial flow of D2O into the old seminal was negligible. We concluded that the function of the primary and seminal roots was to collect water from the lateral roots and transport it to the shoot. A maize root system with lateral roots branching from deep primary and seminal roots would be

  18. Development of a duplex lateral flow assay for simultaneous detection of antibodies against African and Classical swine fever viruses.

    PubMed

    Sastre, Patricia; Pérez, Teresa; Costa, Sofia; Yang, Xiaoping; Räber, Alex; Blome, Sandra; Goller, Katja V; Gallardo, Carmina; Tapia, Istar; García, Julia; Sanz, Antonio; Rueda, Paloma

    2016-09-01

    Classical swine fever (CSF) and African swine fever (ASF) are both highly contagious diseases of domestic pigs and wild boar and are clinically indistinguishable. For both diseases, antibody detection is an integral and crucial part of prevention and control measures. The purpose of our study was to develop and initially validate a duplex pen-side test for simultaneous detection and differentiation of specific antibodies against CSF virus (CSFV) and ASF virus (ASFV). The test was based on the major capsid protein VP72 of ASFV and the structural protein E2 of CSFV, both considered the most immunogenic proteins of these viruses. The performance of the pen-side test was evaluated using a panel of porcine samples consisting of experimental, reference, and field sera, with the latter collected from European farms free of both diseases. The new lateral flow assay was able to detect specific antibodies to ASFV or CSFV, showing good levels of sensitivity and specificity. These preliminary data indicate the potential of the newly developed pen-side test for rapid differential detection of antibodies found in the 2 diseases, which is of particular importance in the field and in front-line laboratories where equipment and skilled personnel are limited and control of ASF and CSF is crucial. © 2016 The Author(s).

  19. 3D flow focusing for microfluidic flow cytometry with ultrasonics

    NASA Astrophysics Data System (ADS)

    Gnyawali, Vaskar; Strohm, Eric M.; Daghighi, Yasaman; van de Vondervoort, Mia; Kolios, Michael C.; Tsai, Scott S. H.

    2015-11-01

    We are developing a flow cytometer that detects unique acoustic signature waves generated from single cells due to interactions between the cells and ultrasound waves. The generated acoustic waves depend on the size and biomechanical properties of the cells and are sufficient for identifying cells in the medium. A microfluidic system capable of focusing cells through a 10 x 10 μm ultrasound beam cross section was developed to facilitate acoustic measurements of single cells. The cells are streamlined in a hydro-dynamically 3D focused flow in a 300 x 300 μm channel made using PDMS. 3D focusing is realized by lateral sheath flows and an inlet needle (inner diameter 100 μm). The accuracy of the 3D flow focusing is measured using a dye and detecting its localization using confocal microscopy. Each flowing cell would be probed by an ultrasound pulse, which has a center frequency of 375 MHz and bandwidth of 250 MHz. The same probe would also be used for recording the scattered waves from the cells, which would be processed to distinguish the physical and biomechanical characteristics of the cells, eventually identifying them. This technique has potential applications in detecting circulating tumor cells, blood cells and blood-related diseases.

  20. High-sensitivity direct analysis of aflatoxins in peanuts and cereal matrices by ultra-performance liquid chromatography with fluorescence detection involving a large volume flow cell.

    PubMed

    Oulkar, Dasharath; Goon, Arnab; Dhanshetty, Manisha; Khan, Zareen; Satav, Sagar; Banerjee, Kaushik

    2018-04-03

    This paper reports a sensitive and cost effective method of analysis for aflatoxins B1, B2, G1 and G2. The sample preparation method was primarily optimised in peanuts, followed by its validation in a range of peanut-processed products and cereal (rice, corn, millets) matrices. Peanut slurry [12.5 g peanut + 12.5 mL water] was extracted with methanol: water (8:2, 100 mL), cleaned through an immunoaffinity column and thereafter measured directly by ultra-performance liquid chromatography-fluorescence (UPLC-FLD) detection, within a chromatographic runtime of 5 minutes. The use of a large volume flow cell in the FLD nullified the requirement of any post-column derivatisation and provided the lowest ever reported limits of quantification of 0.025 for B1 and G1 and 0.01 μg/kg for B2 and G2. The single laboratory validation of the method provided acceptable selectivity, linearity, recovery and precision for reliable quantifications in all the test matrices as well as demonstrated compliance with the EC 401/2006 guidelines for analytical quality control of aflatoxins in foodstuffs.

  1. Gold nanoparticle-based enhanced lateral flow immunoassay for detection of Cronobacter sakazakii in powdered infant formula.

    PubMed

    Pan, Ruili; Jiang, Yujun; Sun, Luhong; Wang, Rui; Zhuang, Kejin; Zhao, Yueming; Wang, Hui; Ali, Md Aslam; Xu, Honghua; Man, Chaoxin

    2018-05-01

    Cronobacter sakazakii is an opportunistic foodborne pathogen that can infect newborns through powdered infant formula (PIF). In this study, we developed a novel enhanced lateral flow immunoassay (LFA) with enhanced sensitivity for detection of C. sakazakii in PIF by the naked eye. The proposed strategy for signal enhancement of the traditional LFA used concentrated gold nanoparticles (AuNP) as the enhancer to conjugate with capture antibodies, which could increase the immobilized capture antibodies concentration at the detection zone to improve capture efficiency. Besides, the detection signal was further amplified by accumulated AuNP as the C. sakazakii labeled with AuNP probes was captured by antibodies conjugated with enhancer at the test line. We also studied the effect of different concentrations of capture antibodies and concentrated AuNP on detection performance, and found that 2.2 mg/mL of capture antibodies and 0.06 nM concentrated AuNP were the optimal combination that could avoid a false-positive signal and maximally amplify the detection signal of the enhanced LFA. Using this strategy, the detection sensitivity of the enhanced LFA was 10 3 cfu/mL and improved 100-fold compared with traditional LFA. The strip was highly specific to C. sakazakii, and the time for detection of C. sakazakii in PIF was shortened by 3 h. In summary, the enhanced LFA developed by the addition of concentrated AuNP as the enhancer can be used as a sensitive, rapid, visual qualitative and point-of-care test method for detecting target analytes. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  2. Motor laterality as an indicator of speech laterality.

    PubMed

    Flowers, Kenneth A; Hudson, John M

    2013-03-01

    The determination of speech laterality, especially where it is anomalous, is both a theoretical issue and a practical problem for brain surgery. Handedness is commonly thought to be related to speech representation, but exactly how is not clearly understood. This investigation analyzed handedness by preference rating and performance on a reliable task of motor laterality in 34 patients undergoing a Wada test, to see if they could provide an indicator of speech laterality. Hand usage preference ratings divided patients into left, right, and mixed in preference. Between-hand differences in movement time on a pegboard task determined motor laterality. Results were correlated (χ2) with speech representation as determined by a standard Wada test. It was found that patients whose between-hand difference in speed on the motor task was small or inconsistent were the ones whose Wada test speech representation was likely to be ambiguous or anomalous, whereas all those with a consistently large between-hand difference showed clear unilateral speech representation in the hemisphere controlling the better hand (χ2 = 10.45, df = 1, p < .01, η2 = 0.55) This relationship prevailed across hand preference and level of skill in the hands itself. We propose that motor and speech laterality are related where they both involve a central control of motor output sequencing and that a measure of that aspect of the former will indicate the likely representation of the latter. A between-hand measure of motor laterality based on such a measure may indicate the possibility of anomalous speech representation. PsycINFO Database Record (c) 2013 APA, all rights reserved.

  3. Lateral flow devices

    DOEpatents

    Mazumdar, Debapriya; Liu, Juewen; Lu, Yi

    2010-09-21

    An analytical test for an analyte comprises (a) a base, having a reaction area and a visualization area, (b) a capture species, on the base in the visualization area, comprising nucleic acid, and (c) analysis chemistry reagents, on the base in the reaction area. The analysis chemistry reagents comprise (i) a substrate comprising nucleic acid and a first label, and (ii) a reactor comprising nucleic acid. The analysis chemistry reagents can react with a sample comprising the analyte and water, to produce a visualization species comprising nucleic acid and the first label, and the capture species can bind the visualization species.

  4. On the Lateral Static Stability of Low-Aspect-Ratio Rectangular Wings

    NASA Astrophysics Data System (ADS)

    Linehan, Thomas; Mohseni, Kamran

    2017-11-01

    Low-aspect-ratio rectangular wings experience a reduction in lateral static stability at angles of attack distinct from that of lift stall. Stereoscopic digital particle image velocimetry is used to elucidate the flow physics behind this trend. Rectangular wings of AR = 0.75, 1, 1.5, 3 were tested at side-slip angles β = -10° and 0° with angle of attack varied in the range α =10° -40° . In side-slip, the leading-edge separation region emerges on the leeward wing where leading-edge flow reattachment is highly intermittent due to vortex shedding. The tip vortex downwash of the AR < 1.5 wings is sufficient to restrict the shedding of leading-edge vorticity, enabling sustained lift from the leading-edge separation region to high angles of attack. The windward tip vortex grows in size with increasing angle of attack, occupying an increasingly larger percentage of the windward wing. At high angles of attack pre-lift stall, the windward tip vortex lifts off the wing, resulting in separated flow underneath it. The downwash of the AR = 3 wing is insufficient to reattach the leading-edge flow at high incidence. The flow stalls on the leeward wing with stalled flow expanding upstream toward the windward wing with increasing angle of attack.

  5. Regimes of Two-Phase Flow in Short Rectangular Channel

    NASA Astrophysics Data System (ADS)

    Chinnov, Evgeny A.; Guzanov, Vladimir V.; Cheverda, Vyacheslav; Markovich, Dmitry M.; Kabov, Oleg A.

    2009-08-01

    Experimental study of two-phase flow in the short rectangular horizontal channel with height 440 μm has been performed. Characteristics of liquid motion inside the channel have been registered and measured by the Laser Induced Fluorescence technique. New information has allowed determining more precisely the characteristics of churn regime and boundaries between different regimes of two-phase flow. It was shown that formation of some two-phase flow regimes and transitions between them are determined by instability of the flow in the lateral parts of the channel.

  6. Reducing the critical particle diameter in (highly) asymmetric sieve-based lateral displacement devices.

    PubMed

    Dijkshoorn, J P; Schutyser, M A I; Sebris, M; Boom, R M; Wagterveld, R M

    2017-10-26

    Deterministic lateral displacement technology was originally developed in the realm of microfluidics, but has potential for larger scale separation as well. In our previous studies, we proposed a sieve-based lateral displacement device inspired on the principle of deterministic lateral displacement. The advantages of this new device is that it gives a lower pressure drop, lower risk of particle accumulation, higher throughput and is simpler to manufacture. However, until now this device has only been investigated for its separation of large particles of around 785 µm diameter. To separate smaller particles, we investigate several design parameters for their influence on the critical particle diameter. In a dimensionless evaluation, device designs with different geometry and dimensions were compared. It was found that sieve-based lateral displacement devices are able to displace particles due to the crucial role of the flow profile, despite of their unusual and asymmetric design. These results demonstrate the possibility to actively steer the velocity profile in order to reduce the critical diameter in deterministic lateral displacement devices, which makes this separation principle more accessible for large-scale, high throughput applications.

  7. Response format, magnitude of laterality effects, and sex differences in laterality.

    PubMed

    Voyer, Daniel; Doyle, Randi A

    2012-01-01

    The present study examined the evidence for the claim that response format might affect the magnitude of laterality effects by means of a meta-analysis. The analysis included the 396 effect sizes drawn from 266 studies retrieved by Voyer (1996) and relevant to the main effect of laterality and sex differences in laterality for verbal and non-verbal tasks in the auditory, tactile, and visual sensory modality. The response format used in specific studies was the only moderator variable of interest in the present analysis, resulting in four broad response categories (oral, written, computer, and pointing). A meta-analysis analogue to ANOVA showed no significant influence of response format on either the main effect of laterality or sex differences in laterality when all sensory modalities were combined. However, when modalities were considered separately, response format affected the main effect of laterality in the visual modality, with a clear advantage for written responses. Further pointed analyses revealed some specific differences among response formats. Results are discussed in terms of their implications for the measurement of laterality.

  8. Autumn Leaf Chromatography.

    ERIC Educational Resources Information Center

    Scharmann, Lawrence C.

    1984-01-01

    Describes an experiment designed to introduce students to chromatographic techniques. Also describes a teacher demonstration in which leaves obtained during the spring and fall are analyzed using chromatography. Procedures for both the experiment and the demonstration are outlined. (JN)

  9. Screening anti-tumor compounds from Ligusticum wallichii using cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry.

    PubMed

    Zhang, Tao; Ding, Yuanyuan; An, Hongli; Feng, Liuxin; Wang, Sicen

    2015-07-14

    Tyrosine 367 Cysteine-fibroblast growth factor receptor 4 cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry was developed. Tyrosine 367 Cysteine-HEK293 cells were used as cell membrane stationary phase. Specificity and reproducibility of the cell membrane chromatography was evaluated using 1-tert-butyl-3-{2-[4-(diethylamino)butylamino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl}urea, Nimodipine and dexamethasone acetate. Then, anti-tumor components acting on Tyrosine 367 Cysteine-fibroblast growth factor receptor 4 were screened and identified from extracts of Ligusticum wallichii. Components from the extract were retained on the cell membrane chromatographic column. The retained fraction was directly eluted into high-performance liquid chromatography with mass spectrometry system for separation and identification. Finally, Levistolide A was identified as an active component from Ligusticum wallichii extracts. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide-formazan colorimetric assay revealed that Levistolide A inhibits proliferation of overexpressing the mutated receptor cells with dose-dependent manner. Phosphorylation of fibroblast growth factor receptor 4 was also decrease under Levistolide A treatment. Flex dock simulation verified that Levistolide A could bind with the tyrosine kinase domain of fibroblast growth factor receptor 4. Therefore, Levistolide A screened by the cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry can arrest cell growth. In conclusion, the two-dimensional high-performance liquid chromatography method can screen and identify potential anti-tumor ingredients which specifically act on the tyrosine kinase domain of the mutated fibroblast growth factor receptor 4. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  10. Longitudinal variation in lateral trapping of fine sediment in tidal estuaries: observations and a 3D exploratory model

    NASA Astrophysics Data System (ADS)

    Chen, Wei; de Swart, Huib E.

    2018-03-01

    This study investigates the longitudinal variation of lateral entrapment of suspended sediment, as is observed in some tidal estuaries. In particular, field data from the Yangtze Estuary are analysed, which reveal that in one cross-section, two maxima of suspended sediment concentration (SSC) occur close to the south and north sides, while in a cross-section 2 km down-estuary, only one SSC maximum on the south side is present. This pattern is found during both spring tide and neap tide, which are characterised by different intensities of turbulence. To understand longitudinal variation in lateral trapping of sediment, results of a new three-dimensional exploratory model are analysed. The hydrodynamic part contains residual flow due to fresh water input, density gradients and Coriolis force and due to channel curvature-induced leakage. Moreover, the model includes a spatially varying eddy viscosity that accounts for variation of intensity of turbulence over the spring-neap cycle. By imposing morphodynamic equilibrium, the two-dimensional distribution of sediment in the domain is obtained analytically by a novel procedure. Results reveal that the occurrence of the SSC maxima near the south side of both cross-sections is due to sediment entrapment by lateral density gradients, while the second SSC maximum near the north side of the first cross-section is by sediment transport due to curvature-induced leakage. Coriolis deflection of longitudinal flow also contributes the trapping of sediment near the north side. This mechanism is important in the upper estuary, where the flow due to lateral density gradients is weak.

  11. Instrument platforms for nano liquid chromatography.

    PubMed

    Šesták, Jozef; Moravcová, Dana; Kahle, Vladislav

    2015-11-20

    The history of liquid chromatography started more than a century ago and miniaturization and automation are two leading trends in this field. Nanocolumn liquid chromatography (nano LC) and largely synonymous capillary liquid chromatography (capillary LC) are the most recent results of this process where miniaturization of column dimensions and sorbent particle size play crucial role. Very interesting results achieved in the research of extremely miniaturized LC columns at the end of the last century lacked distinctive raison d'être and only advances in mass spectrometry brought a real breakthrough. Configuration of nano LC-electrospray ionization mass spectrometry (LC-ESI-MS) has become a basic tool in bioanalytical chemistry, especially in proteomics. This review discusses and summarizes past and current trends in the realization of nano liquid chromatography (nano LC) platforms. Special attention is given to the mobile phase delivery under nanoflow rates (isocratic, gradient) and sample injection to the nanocolumn. Available detection techniques applied in nano LC separations are also briefly discussed. We followed up the key themes from the original scientific reports over gradual improvements up to the contemporary commercial solutions. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. A high-performance liquid chromatography micromethod for the simultaneous determination of vigabatrin and gabapentin in serum.

    PubMed

    Ratnaraj, N; Patsalos, P N

    1998-08-01

    A gradient high-performance liquid chromatography micromethod is described for the simultaneous quantitation of vigabatrin and gabapentin in human serum. Chromatography was performed using a 125- x 3-mm ID Hypersil BDS C-18 column with a 3-microm mini-bore, eluted with a gradient system comprised of phosphate buffer (pH 6.5)-acetonitrile-methanol-water at a flow rate of 0.45 ml/minute. The column eluent was monitored on a fluorescence detector using excitation and emission wavelengths of 340 and 440 nm, respectively. The lower limit of quantitation for vigabatrin and for gabapentin was 5 micromol/l, and the within-batch and between-batch coefficients of variation were <5%. No interference from commonly prescribed antiepileptic drugs (carbamazepine and its metabolite carbamazepine epoxide, oxcarbazepine and its metabolite 10-hydroxycarbazepine, ethosuximide, lamotrigine, phenobarbitone, phenytoin, primidone, and valproic acid) was observed; thus, the method can be used to monitor vigabatrin and gabapentin in patients on polytherapy antiepileptic drug regimens.

  13. Two-dimensional simulation of red blood cell motion near a wall under a lateral force

    NASA Astrophysics Data System (ADS)

    Hariprasad, Daniel S.; Secomb, Timothy W.

    2014-11-01

    The motion of a red blood cell suspended in a linear shear flow adjacent to a fixed boundary subject to an applied lateral force directed toward the boundary is simulated. A two-dimensional model is used that represents the viscous and elastic properties of normal red blood cells. Shear rates in the range of 100 to 600 s-1 are considered, and the suspending medium viscosity is 1 cP. In the absence of a lateral force, the cell executes a tumbling motion. With increasing lateral force, a transition from tumbling to tank-treading is predicted. The minimum force required to ensure tank-treading increases nonlinearly with the shear rate. Transient swinging motions occur when the force is slightly larger than the transition value. The applied lateral force is balanced by a hydrodynamic lift force resulting from the positive orientation of the long axis of the cell with respect to the wall. In the case of cyclic tumbling motions, the orientation angle takes positive values through most of the cycle, resulting in lift generation. These results are used to predict the motion of a cell close to the outer edge of the cell-rich core region that is generated when blood flows in a narrow tube. In this case, the lateral force is generated by shear-induced dispersion, resulting from cell-cell interactions in a region with a concentration gradient. This force is estimated using previous data on shear-induced dispersion. The cell is predicted to execute tank-treading motions at normal physiological hematocrit levels, with the possibility of tumbling at lower hematocrit levels.

  14. Proteomic analysis of exosomes from human neural stem cells by flow field-flow fractionation and nanoflow liquid chromatography-tandem mass spectrometry.

    PubMed

    Kang, Dukjin; Oh, Sunok; Ahn, Sung-Min; Lee, Bong-Hee; Moon, Myeong Hee

    2008-08-01

    Exosomes, small membrane vesicles secreted by a multitude of cell types, are involved in a wide range of physiological roles such as intercellular communication, membrane exchange between cells, and degradation as an alternative to lysosomes. Because of the small size of exosomes (30-100 nm) and the limitations of common separation procedures including ultracentrifugation and flow cytometry, size-based fractionation of exosomes has been challenging. In this study, we used flow field-flow fractionation (FlFFF) to fractionate exosomes according to differences in hydrodynamic diameter. The exosome fractions collected from FlFFF runs were examined by transmission electron microscopy (TEM) to morphologically confirm their identification as exosomes. Exosomal lysates of each fraction were digested and analyzed using nanoflow LC-ESI-MS-MS for protein identification. FIFFF, coupled with mass spectrometry, allows nanoscale size-based fractionation of exosomes and is more applicable to primary cells and stem cells since it requires much less starting material than conventional gel-based separation, in-gel digestion and the MS-MS method.

  15. Experimental Investigation of the Flow Structure over a Delta Wing Via Flow Visualization Methods.

    PubMed

    Shen, Lu; Chen, Zong-Nan; Wen, Chihyung

    2018-04-23

    It is well known that the flow field over a delta wing is dominated by a pair of counter rotating leading edge vortices (LEV). However, their mechanism is not well understood. The flow visualization technique is a promising non-intrusive method to illustrate the complex flow field spatially and temporally. A basic flow visualization setup consists of a high-powered laser and optic lenses to generate the laser sheet, a camera, a tracer particle generator, and a data processor. The wind tunnel setup, the specifications of devices involved, and the corresponding parameter settings are dependent on the flow features to be obtained. Normal smoke wire flow visualization uses a smoke wire to demonstrate the flow streaklines. However, the performance of this method is limited by poor spatial resolution when it is conducted in a complex flow field. Therefore, an improved smoke flow visualization technique has been developed. This technique illustrates the large-scale global LEV flow field and the small-scale shear layer flow structure at the same time, providing a valuable reference for later detailed particle image velocimetry (PIV) measurement. In this paper, the application of the improved smoke flow visualization and PIV measurement to study the unsteady flow phenomena over a delta wing is demonstrated. The procedure and cautions for conducting the experiment are listed, including wind tunnel setup, data acquisition, and data processing. The representative results show that these two flow visualization methods are effective techniques for investigating the three-dimensional flow field qualitatively and quantitatively.

  16. Cell-phone-based measurement of TSH using Mie scatter optimized lateral flow assays.

    PubMed

    You, David J; Park, Tu San; Yoon, Jeong-Yeol

    2013-02-15

    Semi-quantitative thyr oid stimulating hormone (TSH) lateral flow immunochromatographic assays (LFA) are used to screen for serum TSH concentration >5 mIUL(-1) (hypothyroidism). The LFA format, however, is unable to measure TSH in the normal range or detect suppressed levels of TSH (<0.4 mIU L(-1); hyperthyroidism). In fact, it does not provide quantitative TSH values at all. Obtaining quantitative TSH results, especially in the low concentration range, has until now required the use of centralized clinical laboratories which require specimen transport, specialized equipment and personnel, and result in increased cost and delays in the timely reporting of important clinical results. We have conducted a series of experiments to develop and validate an optical system and image analysis algorithm based upon a cell phone platform. It is able to provide point-of-care quantitative TSH results with a high level of sensitivity and reproducibility comparable to that of a clinical laboratory-based third-generation TSH immunoassay. Our research approach uses the methodology of the optimized Rayleigh/Mie scatter detection by taking into consideration the optical characteristics of a nitrocellulose membrane and gold nanoparticles on an LFA for quantifying TSH levels. Using a miniature spectrometer, LED light source, and optical fibers on a rotating benchtop apparatus, the light intensity from different angles of incident light and angles of detection to the LFA were measured. The optimum angles were found that the minimized Mie scattering from nitrocellulose membrane, consequently maximizes the Rayleigh scatter detection from the gold nanoparticles in the LFA bands. Using the results from the benchtop apparatus, a cell-phone-based apparatus was designed which utilized the embedded flash in the cell phone camera as the light source, piped the light with an optical fiber from the flash through a collimating lens to illuminate the LFA. Quantification of TSH was performed in an i

  17. Development of improved enzyme-based and lateral flow immunoassays for rapid and accurate serodiagnosis of canine brucellosis.

    PubMed

    Cortina, María E; Novak, Analía; Melli, Luciano J; Elena, Sebastián; Corbera, Natalia; Romero, Juan E; Nicola, Ana M; Ugalde, Juan E; Comerci, Diego J; Ciocchini, Andrés E

    2017-09-01

    Brucellosis is a widespread zoonotic disease caused by Brucella spp. Brucella canis is the etiological agent of canine brucellosis, a disease that can lead to sterility in bitches and dogs causing important economic losses in breeding kennels. Early and accurate diagnosis of canine brucellosis is central to control the disease and lower the risk of transmission to humans. Here, we develop and validate enzyme and lateral flow immunoassays for improved serodiagnosis of canine brucellosis using as antigen the B. canis rough lipopolysaccharide (rLPS). The method used to obtain the rLPS allowed us to produce more homogeneous batches of the antigen that facilitated the standardization of the assays. To validate the assays, 284 serum samples obtained from naturally infected dogs and healthy animals were analyzed. For the B. canis-iELISA and B. canis-LFIA the diagnostic sensitivity was of 98.6%, and the specificity 99.5% and 100%, respectively. We propose the implementation of the B. canis-LFIA as a screening test in combination with the highly accurate laboratory g-iELISA. The B. canis-LFIA is a rapid, accurate and easy to use test, characteristics that make it ideal for the serological surveillance of canine brucellosis in the field or veterinary laboratories. Finally, a blind study including 1040 serum samples obtained from urban dogs showed a prevalence higher than 5% highlighting the need of new diagnostic tools for a more effective control of the disease in dogs and therefore to reduce the risk of transmission of this zoonotic pathogen to humans. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Comprehensive analysis of pharmaceutical products using simultaneous mixed-mode (ion-exchange/reversed-phase) and hydrophilic interaction liquid chromatography.

    PubMed

    Kazarian, Artaches A; Nesterenko, Pavel N; Soisungnoen, Phimpha; Burakham, Rodjana; Srijaranai, Supalax; Paull, Brett

    2014-08-01

    Liquid chromatographic assays were developed using a mixed-mode column coupled in sequence with a hydrophilic interaction liquid chromatography column to allow the simultaneous comprehensive analysis of inorganic/organic anions and cations, active pharmaceutical ingredients, and excipients (carbohydrates). The approach utilized dual sample injection and valve-mediated column switching and was based upon a single high-performance liquid chromatography gradient pump. The separation consisted of three distinct sequential separation mechanisms, namely, (i) ion-exchange, (ii) mixed-mode interactions under an applied dual gradient (reversed-phase/ion-exchange), and (iii) hydrophilic interaction chromatography. Upon first injection, the Scherzo SS C18 column (Imtakt) provided resolution of inorganic anions and cations under isocratic conditions, followed by a dual organic/salt gradient to elute active pharmaceutical ingredients and their respective organic counterions and potential degradants. At the top of the mixed-mode gradient (high acetonitrile content), the mobile phase flow was switched to a preconditioned hydrophilic interaction liquid chromatography column, and the standard/sample was reinjected for the separation of hydrophilic carbohydrates, some of which are commonly known excipients in drug formulations. The approach afforded reproducible separation and resolution of up to 23 chemically diverse solutes in a single run. The method was applied to investigate the composition of commercial cough syrups (Robitussin®), allowing resolution and determination of inorganic ions, active pharmaceutical ingredients, excipients, and numerous well-resolved unknown peaks. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Ionic liquid stationary phases for gas chromatography.

    PubMed

    Poole, Colin F; Poole, Salwa K

    2011-04-01

    This article provides a summary of the development of ionic liquids as stationary phases for gas chromatography beginning with early work on packed columns that established details of the retention mechanism and established working methods to characterize selectivity differences compared with molecular stationary phases through the modern development of multi-centered cation and cross-linked ionic liquids for high-temperature applications in capillary gas chromatography. Since there are many reviews on ionic liquids dealing with all aspects of their chemical and physical properties, the emphasis in this article is placed on the role of gas chromatography played in the design of ionic liquids of low melting point, high thermal stability, high viscosity, and variable selectivity for separations. Ionic liquids provide unprecedented opportunities for extending the selectivity range and temperature-operating range of columns for gas chromatography, an area of separation science that has otherwise been almost stagnant for over a decade. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Measuring acetabular component position on lateral radiographs - ischio-lateral method.

    PubMed

    Pulos, Nicholas; Tiberi Iii, John V; Schmalzried, Thomas P

    2011-01-01

    The standard method for the evaluation of arthritis and postoperative assessment of arthroplasty treatment is observation and measurement from plain films, using the flm edge for orientation. A more recent employment of an anatomical landmark, the ischial tuberosity, has come into use as orientation for evaluation and is called the ischio-lateral method. In this study, the use of this method was evaluated as a first report to the literature on acetabular component measurement using a skeletal reference with lateral radiographs. Postoperative radiographs of 52 hips, with at least three true lateral radiographs taken at different time periods, were analyzed. Component position was measured with the historical method (using the flm edge for orientation) and with the new method using the ischio-lateral method. The mean standard deviation (SD) for the historical approach was 3.7° and for the ischio-lateral method, 2.2° (p < 0.001). With the historical method, 19 (36.5%) hips had a SD greater than ± 4°, compared to six hips (11.5%) with the ischio-lateral method. By using a skeletal reference, the ischio-lateral method provides a more consistent measurement of acetabular component position. The high intra-class correlation coefficients for both intra- and inter-observer reliability indicate that the angle measured with this simple method, which employs no further technology, increased time, or cost, is consistent and reproducible for multiple observers.