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Sample records for accurate quantitative determination

  1. Accurate determination of human serum transferrin isoforms: Exploring metal-specific isotope dilution analysis as a quantitative proteomic tool.

    PubMed

    Busto, M Estela del Castillo; Montes-Bayón, Maria; Sanz-Medel, Alfredo

    2006-12-15

    Carbohydrate-deficient transferrin (CDT) measurements are considered a reliable marker for chronic alcohol consumption, and its use is becoming extensive in forensic medicine. However, CDT is not a single molecular entity but refers to a group of sialic acid-deficient transferrin isoforms from mono- to trisialotransferrin. Thus, the development of methods to analyze accurately and precisely individual transferrin isoforms in biological fluids such as serum is of increasing importance. The present work illustrates the use of ICPMS isotope dilution analysis for the quantification of transferrin isoforms once saturated with iron and separated by anion exchange chromatography (Mono Q 5/50) using a mobile phase consisting of a gradient of ammonium acetate (0-250 mM) in 25 mM Tris-acetic acid (pH 6.5). Species-specific and species-unspecific spikes have been explored. In the first part of the study, the use of postcolumn addition of a solution of 200 ng mL(-1) isotopically enriched iron (57Fe, 95%) in 25 mM sodium citrate/citric acid (pH 4) permitted the quantification of individual sialoforms of transferrin (from S2 to S5) in human serum samples of healthy individuals as well as alcoholic patients. Second, the species-specific spike method was performed by synthesizing an isotopically enriched standard of saturated transferrin (saturated with 57Fe). The characterization of the spike was performed by postcolumn reverse isotope dilution analysis (this is, by postcolumn addition of a solution of 200 ng mL(-1) natural iron in sodium citrate/citric acid of pH 4). Also, the stability of the transferrin spike was tested during one week with negligible species transformation. Finally, the enriched transferrin was used to quantify the individual isoforms in the same serum samples obtaining results comparative to those of postcolumn isotope dilution and to those previously published in the literature, demonstrating the suitability of both strategies for quantitative transferrin

  2. Analytical method for the accurate determination of tricothecenes in grains using LC-MS/MS: a comparison between MRM transition and MS3 quantitation.

    PubMed

    Lim, Chee Wei; Tai, Siew Hoon; Lee, Lin Min; Chan, Sheot Harn

    2012-07-01

    The current food crisis demands unambiguous determination of mycotoxin contamination in staple foods to achieve safer food for consumption. This paper describes the first accurate LC-MS/MS method developed to analyze tricothecenes in grains by applying multiple reaction monitoring (MRM) transition and MS(3) quantitation strategies in tandem. The tricothecenes are nivalenol, deoxynivalenol, deoxynivalenol-3-glucoside, fusarenon X, 3-acetyl-deoxynivalenol, 15-acetyldeoxynivalenol, diacetoxyscirpenol, and HT-2 and T-2 toxins. Acetic acid and ammonium acetate were used to convert the analytes into their respective acetate adducts and ammonium adducts under negative and positive MS polarity conditions, respectively. The mycotoxins were separated by reversed-phase LC in a 13.5-min run, ionized using electrospray ionization, and detected by tandem mass spectrometry. Analyte-specific mass-to-charge (m/z) ratios were used to perform quantitation under MRM transition and MS(3) (linear ion trap) modes. Three experiments were made for each quantitation mode and matrix in batches over 6 days for recovery studies. The matrix effect was investigated at concentration levels of 20, 40, 80, 120, 160, and 200 μg kg(-1) (n = 3) in 5 g corn flour and rice flour. Extraction with acetonitrile provided a good overall recovery range of 90-108% (n = 3) at three levels of spiking concentration of 40, 80, and 120 μg kg(-1). A quantitation limit of 2-6 μg kg(-1) was achieved by applying an MRM transition quantitation strategy. Under MS(3) mode, a quantitation limit of 4-10 μg kg(-1) was achieved. Relative standard deviations of 2-10% and 2-11% were reported for MRM transition and MS(3) quantitation, respectively. The successful utilization of MS(3) enabled accurate analyte fragmentation pattern matching and its quantitation, leading to the development of analytical methods in fields that demand both analyte specificity and fragmentation fingerprint-matching capabilities that are

  3. Toward Accurate and Quantitative Comparative Metagenomics.

    PubMed

    Nayfach, Stephen; Pollard, Katherine S

    2016-08-25

    Shotgun metagenomics and computational analysis are used to compare the taxonomic and functional profiles of microbial communities. Leveraging this approach to understand roles of microbes in human biology and other environments requires quantitative data summaries whose values are comparable across samples and studies. Comparability is currently hampered by the use of abundance statistics that do not estimate a meaningful parameter of the microbial community and biases introduced by experimental protocols and data-cleaning approaches. Addressing these challenges, along with improving study design, data access, metadata standardization, and analysis tools, will enable accurate comparative metagenomics. We envision a future in which microbiome studies are replicable and new metagenomes are easily and rapidly integrated with existing data. Only then can the potential of metagenomics for predictive ecological modeling, well-powered association studies, and effective microbiome medicine be fully realized. PMID:27565341

  4. Toward Accurate and Quantitative Comparative Metagenomics

    PubMed Central

    Nayfach, Stephen; Pollard, Katherine S.

    2016-01-01

    Shotgun metagenomics and computational analysis are used to compare the taxonomic and functional profiles of microbial communities. Leveraging this approach to understand roles of microbes in human biology and other environments requires quantitative data summaries whose values are comparable across samples and studies. Comparability is currently hampered by the use of abundance statistics that do not estimate a meaningful parameter of the microbial community and biases introduced by experimental protocols and data-cleaning approaches. Addressing these challenges, along with improving study design, data access, metadata standardization, and analysis tools, will enable accurate comparative metagenomics. We envision a future in which microbiome studies are replicable and new metagenomes are easily and rapidly integrated with existing data. Only then can the potential of metagenomics for predictive ecological modeling, well-powered association studies, and effective microbiome medicine be fully realized. PMID:27565341

  5. Accurate SHAPE-directed RNA structure determination

    PubMed Central

    Deigan, Katherine E.; Li, Tian W.; Mathews, David H.; Weeks, Kevin M.

    2009-01-01

    Almost all RNAs can fold to form extensive base-paired secondary structures. Many of these structures then modulate numerous fundamental elements of gene expression. Deducing these structure–function relationships requires that it be possible to predict RNA secondary structures accurately. However, RNA secondary structure prediction for large RNAs, such that a single predicted structure for a single sequence reliably represents the correct structure, has remained an unsolved problem. Here, we demonstrate that quantitative, nucleotide-resolution information from a SHAPE experiment can be interpreted as a pseudo-free energy change term and used to determine RNA secondary structure with high accuracy. Free energy minimization, by using SHAPE pseudo-free energies, in conjunction with nearest neighbor parameters, predicts the secondary structure of deproteinized Escherichia coli 16S rRNA (>1,300 nt) and a set of smaller RNAs (75–155 nt) with accuracies of up to 96–100%, which are comparable to the best accuracies achievable by comparative sequence analysis. PMID:19109441

  6. Challenges in accurate quantitation of lysophosphatidic acids in human biofluids

    PubMed Central

    Onorato, Joelle M.; Shipkova, Petia; Minnich, Anne; Aubry, Anne-Françoise; Easter, John; Tymiak, Adrienne

    2014-01-01

    Lysophosphatidic acids (LPAs) are biologically active signaling molecules involved in the regulation of many cellular processes and have been implicated as potential mediators of fibroblast recruitment to the pulmonary airspace, pointing to possible involvement of LPA in the pathology of pulmonary fibrosis. LPAs have been measured in various biological matrices and many challenges involved with their analyses have been documented. However, little published information is available describing LPA levels in human bronchoalveolar lavage fluid (BALF). We therefore conducted detailed investigations into the effects of extensive sample handling and sample preparation conditions on LPA levels in human BALF. Further, targeted lipid profiling of human BALF and plasma identified the most abundant lysophospholipids likely to interfere with LPA measurements. We present the findings from these investigations, highlighting the importance of well-controlled sample handling for the accurate quantitation of LPA. Further, we show that chromatographic separation of individual LPA species from their corresponding lysophospholipid species is critical to avoid reporting artificially elevated levels. The optimized sample preparation and LC/MS/MS method was qualified using a stable isotope-labeled LPA as a surrogate calibrant and used to determine LPA levels in human BALF and plasma from a Phase 0 clinical study comparing idiopathic pulmonary fibrosis patients to healthy controls. PMID:24872406

  7. Groundtruth approach to accurate quantitation of fluorescence microarrays

    SciTech Connect

    Mascio-Kegelmeyer, L; Tomascik-Cheeseman, L; Burnett, M S; van Hummelen, P; Wyrobek, A J

    2000-12-01

    To more accurately measure fluorescent signals from microarrays, we calibrated our acquisition and analysis systems by using groundtruth samples comprised of known quantities of red and green gene-specific DNA probes hybridized to cDNA targets. We imaged the slides with a full-field, white light CCD imager and analyzed them with our custom analysis software. Here we compare, for multiple genes, results obtained with and without preprocessing (alignment, color crosstalk compensation, dark field subtraction, and integration time). We also evaluate the accuracy of various image processing and analysis techniques (background subtraction, segmentation, quantitation and normalization). This methodology calibrates and validates our system for accurate quantitative measurement of microarrays. Specifically, we show that preprocessing the images produces results significantly closer to the known ground-truth for these samples.

  8. Fast and Accurate Detection of Multiple Quantitative Trait Loci

    PubMed Central

    Nettelblad, Carl; Holmgren, Sverker

    2013-01-01

    Abstract We present a new computational scheme that enables efficient and reliable quantitative trait loci (QTL) scans for experimental populations. Using a standard brute-force exhaustive search effectively prohibits accurate QTL scans involving more than two loci to be performed in practice, at least if permutation testing is used to determine significance. Some more elaborate global optimization approaches, for example, DIRECT have been adopted earlier to QTL search problems. Dramatic speedups have been reported for high-dimensional scans. However, since a heuristic termination criterion must be used in these types of algorithms, the accuracy of the optimization process cannot be guaranteed. Indeed, earlier results show that a small bias in the significance thresholds is sometimes introduced. Our new optimization scheme, PruneDIRECT, is based on an analysis leading to a computable (Lipschitz) bound on the slope of a transformed objective function. The bound is derived for both infinite- and finite-size populations. Introducing a Lipschitz bound in DIRECT leads to an algorithm related to classical Lipschitz optimization. Regions in the search space can be permanently excluded (pruned) during the optimization process. Heuristic termination criteria can thus be avoided. Hence, PruneDIRECT has a well-defined error bound and can in practice be guaranteed to be equivalent to a corresponding exhaustive search. We present simulation results that show that for simultaneous mapping of three QTLS using permutation testing, PruneDIRECT is typically more than 50 times faster than exhaustive search. The speedup is higher for stronger QTL. This could be used to quickly detect strong candidate eQTL networks. PMID:23919387

  9. Accurate determination of characteristic relative permeability curves

    NASA Astrophysics Data System (ADS)

    Krause, Michael H.; Benson, Sally M.

    2015-09-01

    A recently developed technique to accurately characterize sub-core scale heterogeneity is applied to investigate the factors responsible for flowrate-dependent effective relative permeability curves measured on core samples in the laboratory. The dependency of laboratory measured relative permeability on flowrate has long been both supported and challenged by a number of investigators. Studies have shown that this apparent flowrate dependency is a result of both sub-core scale heterogeneity and outlet boundary effects. However this has only been demonstrated numerically for highly simplified models of porous media. In this paper, flowrate dependency of effective relative permeability is demonstrated using two rock cores, a Berea Sandstone and a heterogeneous sandstone from the Otway Basin Pilot Project in Australia. Numerical simulations of steady-state coreflooding experiments are conducted at a number of injection rates using a single set of input characteristic relative permeability curves. Effective relative permeability is then calculated from the simulation data using standard interpretation methods for calculating relative permeability from steady-state tests. Results show that simplified approaches may be used to determine flowrate-independent characteristic relative permeability provided flow rate is sufficiently high, and the core heterogeneity is relatively low. It is also shown that characteristic relative permeability can be determined at any typical flowrate, and even for geologically complex models, when using accurate three-dimensional models.

  10. Accurate scoring of non-uniform sampling schemes for quantitative NMR

    PubMed Central

    Aoto, Phillip C.; Fenwick, R. Bryn; Kroon, Gerard J. A.; Wright, Peter E.

    2014-01-01

    Non-uniform sampling (NUS) in NMR spectroscopy is a recognized and powerful tool to minimize acquisition time. Recent advances in reconstruction methodologies are paving the way for the use of NUS in quantitative applications, where accurate measurement of peak intensities is crucial. The presence or absence of NUS artifacts in reconstructed spectra ultimately determines the success of NUS in quantitative NMR. The quality of reconstructed spectra from NUS acquired data is dependent upon the quality of the sampling scheme. Here we demonstrate that the best performing sampling schemes make up a very small percentage of the total randomly generated schemes. A scoring method is found to accurately predict the quantitative similarity between reconstructed NUS spectra and those of fully sampled spectra. We present an easy-to-use protocol to batch generate and rank optimal Poisson-gap NUS schedules for use with 2D NMR with minimized noise and accurate signal reproduction, without the need for the creation of synthetic spectra. PMID:25063954

  11. Active contour approach for accurate quantitative airway analysis

    NASA Astrophysics Data System (ADS)

    Odry, Benjamin L.; Kiraly, Atilla P.; Slabaugh, Greg G.; Novak, Carol L.; Naidich, David P.; Lerallut, Jean-Francois

    2008-03-01

    Chronic airway disease causes structural changes in the lungs including peribronchial thickening and airway dilatation. Multi-detector computed tomography (CT) yields detailed near-isotropic images of the lungs, and thus the potential to obtain quantitative measurements of lumen diameter and airway wall thickness. Such measurements would allow standardized assessment, and physicians to diagnose and locate airway abnormalities, adapt treatment, and monitor progress over time. However, due to the sheer number of airways per patient, systematic analysis is infeasible in routine clinical practice without automation. We have developed an automated and real-time method based on active contours to estimate both airway lumen and wall dimensions; the method does not require manual contour initialization but only a starting point on the targeted airway. While the lumen contour segmentation is purely region-based, the estimation of the outer diameter considers the inner wall segmentation as well as local intensity variation, in order anticipate the presence of nearby arteries and exclude them. These properties make the method more robust than the Full-Width Half Maximum (FWHM) approach. Results are demonstrated on a phantom dataset with known dimensions and on a human dataset where the automated measurements are compared against two human operators. The average error on the phantom measurements was 0.10mm and 0.14mm for inner and outer diameters, showing sub-voxel accuracy. Similarly, the mean variation from the average manual measurement was 0.14mm and 0.18mm for inner and outer diameters respectively.

  12. Accurate quantitation of MHC-bound peptides by application of isotopically labeled peptide MHC complexes.

    PubMed

    Hassan, Chopie; Kester, Michel G D; Oudgenoeg, Gideon; de Ru, Arnoud H; Janssen, George M C; Drijfhout, Jan W; Spaapen, Robbert M; Jiménez, Connie R; Heemskerk, Mirjam H M; Falkenburg, J H Frederik; van Veelen, Peter A

    2014-09-23

    Knowledge of the accurate copy number of HLA class I presented ligands is important in fundamental and clinical immunology. Currently, the best copy number determinations are based on mass spectrometry, employing single reaction monitoring (SRM) in combination with a known amount of isotopically labeled peptide. The major drawback of this approach is that the losses during sample pretreatment, i.e. immunopurification and filtration steps, are not well defined and must, therefore, be estimated. In addition, such losses can vary for individual peptides. Therefore, we developed a new approach in which isotopically labeled peptide-MHC monomers (hpMHC) are prepared and added directly after cell lysis, i.e. before the usual sample processing. Using this approach, all losses during sample processing can be accounted for and allows accurate determination of specific MHC class I-presented ligands. Our study pinpoints the immunopurification step as the origin of the rather extreme losses during sample pretreatment and offers a solution to account for these losses. Obviously, this has important implications for accurate HLA-ligand quantitation. The strategy presented here can be used to obtain a reliable view of epitope copy number and thus allows improvement of vaccine design and strategies for immunotherapy.

  13. Accurate Construction of Photoactivated Localization Microscopy (PALM) Images for Quantitative Measurements

    PubMed Central

    Coltharp, Carla; Kessler, Rene P.; Xiao, Jie

    2012-01-01

    Localization-based superresolution microscopy techniques such as Photoactivated Localization Microscopy (PALM) and Stochastic Optical Reconstruction Microscopy (STORM) have allowed investigations of cellular structures with unprecedented optical resolutions. One major obstacle to interpreting superresolution images, however, is the overcounting of molecule numbers caused by fluorophore photoblinking. Using both experimental and simulated images, we determined the effects of photoblinking on the accurate reconstruction of superresolution images and on quantitative measurements of structural dimension and molecule density made from those images. We found that structural dimension and relative density measurements can be made reliably from images that contain photoblinking-related overcounting, but accurate absolute density measurements, and consequently faithful representations of molecule counts and positions in cellular structures, require the application of a clustering algorithm to group localizations that originate from the same molecule. We analyzed how applying a simple algorithm with different clustering thresholds (tThresh and dThresh) affects the accuracy of reconstructed images, and developed an easy method to select optimal thresholds. We also identified an empirical criterion to evaluate whether an imaging condition is appropriate for accurate superresolution image reconstruction with the clustering algorithm. Both the threshold selection method and imaging condition criterion are easy to implement within existing PALM clustering algorithms and experimental conditions. The main advantage of our method is that it generates a superresolution image and molecule position list that faithfully represents molecule counts and positions within a cellular structure, rather than only summarizing structural properties into ensemble parameters. This feature makes it particularly useful for cellular structures of heterogeneous densities and irregular geometries, and

  14. Accurate determination of the amino acid content of selected feedstuffs.

    PubMed

    Rutherfurd, Shane M

    2009-01-01

    The accurate determination of the amino acid content is important. In the present study, a least-squares non-linear regression model of the amino acid content determined over multiple hydrolysis times was used to accurately determine the content of amino acids in five different feedstuffs. These values were compared with 24-h hydrolysis values determined for the same feedstuffs. Overall, approximately two-thirds of the amino acids determined in this study (aspartic acid, threonine, glutamic acid, proline, glycine, alanine, leucine, tyrosine, phenylalanine and arginine) using 24-h hydrolysis were in good agreement (<3% difference). When examined across feedstuffs, the concentration of serine was underestimated by the 24-h hydrolysis method by 4.8%, while the concentrations of histidine and lysine were overestimated by 3.9% and 3.1%, respectively.

  15. Precise and Accurate Density Determination of Explosives Using Hydrostatic Weighing

    SciTech Connect

    B. Olinger

    2005-07-01

    Precise and accurate density determination requires weight measurements in air and water using sufficiently precise analytical balances, knowledge of the densities of air and water, knowledge of thermal expansions, availability of a density standard, and a method to estimate the time to achieve thermal equilibrium with water. Density distributions in pressed explosives are inferred from the densities of elements from a central slice.

  16. Multimodal Quantitative Phase Imaging with Digital Holographic Microscopy Accurately Assesses Intestinal Inflammation and Epithelial Wound Healing.

    PubMed

    Lenz, Philipp; Brückner, Markus; Ketelhut, Steffi; Heidemann, Jan; Kemper, Björn; Bettenworth, Dominik

    2016-01-01

    The incidence of inflammatory bowel disease, i.e., Crohn's disease and Ulcerative colitis, has significantly increased over the last decade. The etiology of IBD remains unknown and current therapeutic strategies are based on the unspecific suppression of the immune system. The development of treatments that specifically target intestinal inflammation and epithelial wound healing could significantly improve management of IBD, however this requires accurate detection of inflammatory changes. Currently, potential drug candidates are usually evaluated using animal models in vivo or with cell culture based techniques in vitro. Histological examination usually requires the cells or tissues of interest to be stained, which may alter the sample characteristics and furthermore, the interpretation of findings can vary by investigator expertise. Digital holographic microscopy (DHM), based on the detection of optical path length delay, allows stain-free quantitative phase contrast imaging. This allows the results to be directly correlated with absolute biophysical parameters. We demonstrate how measurement of changes in tissue density with DHM, based on refractive index measurement, can quantify inflammatory alterations, without staining, in different layers of colonic tissue specimens from mice and humans with colitis. Additionally, we demonstrate continuous multimodal label-free monitoring of epithelial wound healing in vitro, possible using DHM through the simple automated determination of the wounded area and simultaneous determination of morphological parameters such as dry mass and layer thickness of migrating cells. In conclusion, DHM represents a valuable, novel and quantitative tool for the assessment of intestinal inflammation with absolute values for parameters possible, simplified quantification of epithelial wound healing in vitro and therefore has high potential for translational diagnostic use. PMID:27685659

  17. Accurate Method for Determining Adhesion of Cantilever Beams

    SciTech Connect

    Michalske, T.A.; de Boer, M.P.

    1999-01-08

    Using surface micromachined samples, we demonstrate the accurate measurement of cantilever beam adhesion by using test structures which are adhered over long attachment lengths. We show that this configuration has a deep energy well, such that a fracture equilibrium is easily reached. When compared to the commonly used method of determining the shortest attached beam, the present method is much less sensitive to variations in surface topography or to details of capillary drying.

  18. A Quantitative Gas Chromatographic Ethanol Determination.

    ERIC Educational Resources Information Center

    Leary, James J.

    1983-01-01

    Describes a gas chromatographic experiment for the quantitative determination of volume percent ethanol in water ethanol solutions. Background information, procedures, and typical results are included. Accuracy and precision of results are both on the order of two percent. (JN)

  19. FANSe: an accurate algorithm for quantitative mapping of large scale sequencing reads

    PubMed Central

    Zhang, Gong; Fedyunin, Ivan; Kirchner, Sebastian; Xiao, Chuanle; Valleriani, Angelo; Ignatova, Zoya

    2012-01-01

    The most crucial step in data processing from high-throughput sequencing applications is the accurate and sensitive alignment of the sequencing reads to reference genomes or transcriptomes. The accurate detection of insertions and deletions (indels) and errors introduced by the sequencing platform or by misreading of modified nucleotides is essential for the quantitative processing of the RNA-based sequencing (RNA-Seq) datasets and for the identification of genetic variations and modification patterns. We developed a new, fast and accurate algorithm for nucleic acid sequence analysis, FANSe, with adjustable mismatch allowance settings and ability to handle indels to accurately and quantitatively map millions of reads to small or large reference genomes. It is a seed-based algorithm which uses the whole read information for mapping and high sensitivity and low ambiguity are achieved by using short and non-overlapping reads. Furthermore, FANSe uses hotspot score to prioritize the processing of highly possible matches and implements modified Smith–Watermann refinement with reduced scoring matrix to accelerate the calculation without compromising its sensitivity. The FANSe algorithm stably processes datasets from various sequencing platforms, masked or unmasked and small or large genomes. It shows a remarkable coverage of low-abundance mRNAs which is important for quantitative processing of RNA-Seq datasets. PMID:22379138

  20. Accurate fluorescence quantum yield determination by fluorescence correlation spectroscopy.

    PubMed

    Kempe, Daryan; Schöne, Antonie; Fitter, Jörg; Gabba, Matteo

    2015-04-01

    Here, we present a comparative method for the accurate determination of fluorescence quantum yields (QYs) by fluorescence correlation spectroscopy. By exploiting the high sensitivity of single-molecule spectroscopy, we obtain the QYs of samples in the microliter range and at (sub)nanomolar concentrations. Additionally, in combination with fluorescence lifetime measurements, our method allows the quantification of both static and collisional quenching constants. Thus, besides being simple and fast, our method opens up the possibility to photophysically characterize labeled biomolecules under application-relevant conditions and with low sample consumption, which is often important in single-molecule studies.

  1. A correlative imaging based methodology for accurate quantitative assessment of bone formation in additive manufactured implants.

    PubMed

    Geng, Hua; Todd, Naomi M; Devlin-Mullin, Aine; Poologasundarampillai, Gowsihan; Kim, Taek Bo; Madi, Kamel; Cartmell, Sarah; Mitchell, Christopher A; Jones, Julian R; Lee, Peter D

    2016-06-01

    A correlative imaging methodology was developed to accurately quantify bone formation in the complex lattice structure of additive manufactured implants. Micro computed tomography (μCT) and histomorphometry were combined, integrating the best features from both, while demonstrating the limitations of each imaging modality. This semi-automatic methodology registered each modality using a coarse graining technique to speed the registration of 2D histology sections to high resolution 3D μCT datasets. Once registered, histomorphometric qualitative and quantitative bone descriptors were directly correlated to 3D quantitative bone descriptors, such as bone ingrowth and bone contact. The correlative imaging allowed the significant volumetric shrinkage of histology sections to be quantified for the first time (~15 %). This technique demonstrated the importance of location of the histological section, demonstrating that up to a 30 % offset can be introduced. The results were used to quantitatively demonstrate the effectiveness of 3D printed titanium lattice implants.

  2. Accurate and reproducible determination of lignin molar mass by acetobromination.

    PubMed

    Asikkala, Janne; Tamminen, Tarja; Argyropoulos, Dimitris S

    2012-09-12

    The accurate and reproducible determination of lignin molar mass by using size exclusion chromatography (SEC) is challenging. The lignin association effects, known to dominate underivatized lignins, have been thoroughly addressed by reaction with acetyl bromide in an excess of glacial acetic acid. The combination of a concerted acetylation with the introduction of bromine within the lignin alkyl side chains is thought to be responsible for the observed excellent solubilization characteristics acetobromination imparts to a variety of lignin samples. The proposed methodology was compared and contrasted to traditional lignin derivatization methods. In addition, side reactions that could possibly be induced under the acetobromination conditions were explored with native softwood (milled wood lignin, MWL) and technical (kraft) lignin. These efforts lend support toward the use of room temperature acetobromination being a facile, effective, and universal lignin derivatization medium proposed to be employed prior to SEC measurements. PMID:22870925

  3. Accurate determination of segmented X-ray detector geometry

    PubMed Central

    Yefanov, Oleksandr; Mariani, Valerio; Gati, Cornelius; White, Thomas A.; Chapman, Henry N.; Barty, Anton

    2015-01-01

    Recent advances in X-ray detector technology have resulted in the introduction of segmented detectors composed of many small detector modules tiled together to cover a large detection area. Due to mechanical tolerances and the desire to be able to change the module layout to suit the needs of different experiments, the pixels on each module might not align perfectly on a regular grid. Several detectors are designed to permit detector sub-regions (or modules) to be moved relative to each other for different experiments. Accurate determination of the location of detector elements relative to the beam-sample interaction point is critical for many types of experiment, including X-ray crystallography, coherent diffractive imaging (CDI), small angle X-ray scattering (SAXS) and spectroscopy. For detectors with moveable modules, the relative positions of pixels are no longer fixed, necessitating the development of a simple procedure to calibrate detector geometry after reconfiguration. We describe a simple and robust method for determining the geometry of segmented X-ray detectors using measurements obtained by serial crystallography. By comparing the location of observed Bragg peaks to the spot locations predicted from the crystal indexing procedure, the position, rotation and distance of each module relative to the interaction region can be refined. We show that the refined detector geometry greatly improves the results of experiments. PMID:26561117

  4. Accurate determination of segmented X-ray detector geometry.

    PubMed

    Yefanov, Oleksandr; Mariani, Valerio; Gati, Cornelius; White, Thomas A; Chapman, Henry N; Barty, Anton

    2015-11-01

    Recent advances in X-ray detector technology have resulted in the introduction of segmented detectors composed of many small detector modules tiled together to cover a large detection area. Due to mechanical tolerances and the desire to be able to change the module layout to suit the needs of different experiments, the pixels on each module might not align perfectly on a regular grid. Several detectors are designed to permit detector sub-regions (or modules) to be moved relative to each other for different experiments. Accurate determination of the location of detector elements relative to the beam-sample interaction point is critical for many types of experiment, including X-ray crystallography, coherent diffractive imaging (CDI), small angle X-ray scattering (SAXS) and spectroscopy. For detectors with moveable modules, the relative positions of pixels are no longer fixed, necessitating the development of a simple procedure to calibrate detector geometry after reconfiguration. We describe a simple and robust method for determining the geometry of segmented X-ray detectors using measurements obtained by serial crystallography. By comparing the location of observed Bragg peaks to the spot locations predicted from the crystal indexing procedure, the position, rotation and distance of each module relative to the interaction region can be refined. We show that the refined detector geometry greatly improves the results of experiments.

  5. Accurate determination of segmented X-ray detector geometry.

    PubMed

    Yefanov, Oleksandr; Mariani, Valerio; Gati, Cornelius; White, Thomas A; Chapman, Henry N; Barty, Anton

    2015-11-01

    Recent advances in X-ray detector technology have resulted in the introduction of segmented detectors composed of many small detector modules tiled together to cover a large detection area. Due to mechanical tolerances and the desire to be able to change the module layout to suit the needs of different experiments, the pixels on each module might not align perfectly on a regular grid. Several detectors are designed to permit detector sub-regions (or modules) to be moved relative to each other for different experiments. Accurate determination of the location of detector elements relative to the beam-sample interaction point is critical for many types of experiment, including X-ray crystallography, coherent diffractive imaging (CDI), small angle X-ray scattering (SAXS) and spectroscopy. For detectors with moveable modules, the relative positions of pixels are no longer fixed, necessitating the development of a simple procedure to calibrate detector geometry after reconfiguration. We describe a simple and robust method for determining the geometry of segmented X-ray detectors using measurements obtained by serial crystallography. By comparing the location of observed Bragg peaks to the spot locations predicted from the crystal indexing procedure, the position, rotation and distance of each module relative to the interaction region can be refined. We show that the refined detector geometry greatly improves the results of experiments. PMID:26561117

  6. Quantitative determination of atmospheric hydroperoxyl radical

    DOEpatents

    Springston, Stephen R.; Lloyd, Judith; Zheng, Jun

    2007-10-23

    A method for the quantitative determination of atmospheric hydroperoxyl radical comprising: (a) contacting a liquid phase atmospheric sample with a chemiluminescent compound which luminesces on contact with hydroperoxyl radical; (b) determining luminescence intensity from the liquid phase atmospheric sample; and (c) comparing said luminescence intensity from the liquid phase atmospheric sample to a standard luminescence intensity for hydroperoxyl radical. An apparatus for automating the method is also included.

  7. A fluorescence-based quantitative real-time PCR assay for accurate Pocillopora damicornis species identification

    NASA Astrophysics Data System (ADS)

    Thomas, Luke; Stat, Michael; Evans, Richard D.; Kennington, W. Jason

    2016-09-01

    Pocillopora damicornis is one of the most extensively studied coral species globally, but high levels of phenotypic plasticity within the genus make species identification based on morphology alone unreliable. As a result, there is a compelling need to develop cheap and time-effective molecular techniques capable of accurately distinguishing P. damicornis from other congeneric species. Here, we develop a fluorescence-based quantitative real-time PCR (qPCR) assay to genotype a single nucleotide polymorphism that accurately distinguishes P. damicornis from other morphologically similar Pocillopora species. We trial the assay across colonies representing multiple Pocillopora species and then apply the assay to screen samples of Pocillopora spp. collected at regional scales along the coastline of Western Australia. This assay offers a cheap and time-effective alternative to Sanger sequencing and has broad applications including studies on gene flow, dispersal, recruitment and physiological thresholds of P. damicornis.

  8. Quantitative proteomics using the high resolution accurate mass capabilities of the quadrupole-orbitrap mass spectrometer.

    PubMed

    Gallien, Sebastien; Domon, Bruno

    2014-08-01

    High resolution/accurate mass hybrid mass spectrometers have considerably advanced shotgun proteomics and the recent introduction of fast sequencing capabilities has expanded its use for targeted approaches. More specifically, the quadrupole-orbitrap instrument has a unique configuration and its new features enable a wide range of experiments. An overview of the analytical capabilities of this instrument is presented, with a focus on its application to quantitative analyses. The high resolution, the trapping capability and the versatility of the instrument have allowed quantitative proteomic workflows to be redefined and new data acquisition schemes to be developed. The initial proteomic applications have shown an improvement of the analytical performance. However, as quantification relies on ion trapping, instead of ion beam, further refinement of the technique can be expected.

  9. Quantitative real-time PCR for rapid and accurate titration of recombinant baculovirus particles.

    PubMed

    Hitchman, Richard B; Siaterli, Evangelia A; Nixon, Clare P; King, Linda A

    2007-03-01

    We describe the use of quantitative PCR (QPCR) to titer recombinant baculoviruses. Custom primers and probe were designed to gp64 and used to calculate a standard curve of QPCR derived titers from dilutions of a previously titrated baculovirus stock. Each dilution was titrated by both plaque assay and QPCR, producing a consistent and reproducible inverse relationship between C(T) and plaque forming units per milliliter. No significant difference was observed between titers produced by QPCR and plaque assay for 12 recombinant viruses, confirming the validity of this technique as a rapid and accurate method of baculovirus titration.

  10. Nanoparticle counting: towards accurate determination of the molar concentration.

    PubMed

    Shang, Jing; Gao, Xiaohu

    2014-11-01

    Innovations in nanotechnology have brought tremendous opportunities for the advancement of many research frontiers, ranging from electronics, photonics, energy, to medicine. To maximize the benefits of nano-scaled materials in different devices and systems, precise control of their concentration is a prerequisite. While concentrations of nanoparticles have been provided in other forms (e.g., mass), accurate determination of molar concentration, arguably the most useful one for chemical reactions and applications, has been a major challenge (especially for nanoparticles smaller than 30 nm). Towards this significant yet chronic problem, a variety of strategies are currently under development. Most of these strategies are applicable to a specialized group of nanoparticles due to their restrictions on the composition and size range of nanoparticles. As research and uses of nanomaterials are being explored in an unprecedented speed, it is necessary to develop universal strategies that are easy to use and are compatible with nanoparticles of different sizes, compositions, and shapes. This review outlines the theories and applications of current strategies to measure nanoparticle molar concentration, discusses the advantages and limitations of these methods, and provides insights into future directions.

  11. Quantitation and accurate mass analysis of pesticides in vegetables by LC/TOF-MS.

    PubMed

    Ferrer, Imma; Thurman, E Michael; Fernández-Alba, Amadeo R

    2005-05-01

    A quantitative method consisting of solvent extraction followed by liquid chromatography/time-of-flight mass spectrometry (LC/TOF-MS) analysis was developed for the identification and quantitation of three chloronicotinyl pesticides (imidacloprid, acetamiprid, thiacloprid) commonly used on salad vegetables. Accurate mass measurements within 3 ppm error were obtained for all the pesticides studied in various vegetable matrixes (cucumber, tomato, lettuce, pepper), which allowed an unequivocal identification of the target pesticides. Calibration curves covering 2 orders of magnitude were linear over the concentration range studied, thus showing the quantitative ability of TOF-MS as a monitoring tool for pesticides in vegetables. Matrix effects were also evaluated using matrix-matched standards showing no significant interferences between matrixes and clean extracts. Intraday reproducibility was 2-3% relative standard deviation (RSD) and interday values were 5% RSD. The precision (standard deviation) of the mass measurements was evaluated and it was less than 0.23 mDa between days. Detection limits of the chloronicotinyl insecticides in salad vegetables ranged from 0.002 to 0.01 mg/kg. These concentrations are equal to or better than the EU directives for controlled pesticides in vegetables showing that LC/TOF-MS analysis is a powerful tool for identification of pesticides in vegetables. Robustness and applicability of the method was validated for the analysis of market vegetable samples. Concentrations found in these samples were in the range of 0.02-0.17 mg/kg of vegetable. PMID:15859598

  12. Accurate and quantitative polarization-sensitive OCT by unbiased birefringence estimator with noise-stochastic correction

    NASA Astrophysics Data System (ADS)

    Kasaragod, Deepa; Sugiyama, Satoshi; Ikuno, Yasushi; Alonso-Caneiro, David; Yamanari, Masahiro; Fukuda, Shinichi; Oshika, Tetsuro; Hong, Young-Joo; Li, En; Makita, Shuichi; Miura, Masahiro; Yasuno, Yoshiaki

    2016-03-01

    Polarization sensitive optical coherence tomography (PS-OCT) is a functional extension of OCT that contrasts the polarization properties of tissues. It has been applied to ophthalmology, cardiology, etc. Proper quantitative imaging is required for a widespread clinical utility. However, the conventional method of averaging to improve the signal to noise ratio (SNR) and the contrast of the phase retardation (or birefringence) images introduce a noise bias offset from the true value. This bias reduces the effectiveness of birefringence contrast for a quantitative study. Although coherent averaging of Jones matrix tomography has been widely utilized and has improved the image quality, the fundamental limitation of nonlinear dependency of phase retardation and birefringence to the SNR was not overcome. So the birefringence obtained by PS-OCT was still not accurate for a quantitative imaging. The nonlinear effect of SNR to phase retardation and birefringence measurement was previously formulated in detail for a Jones matrix OCT (JM-OCT) [1]. Based on this, we had developed a maximum a-posteriori (MAP) estimator and quantitative birefringence imaging was demonstrated [2]. However, this first version of estimator had a theoretical shortcoming. It did not take into account the stochastic nature of SNR of OCT signal. In this paper, we present an improved version of the MAP estimator which takes into account the stochastic property of SNR. This estimator uses a probability distribution function (PDF) of true local retardation, which is proportional to birefringence, under a specific set of measurements of the birefringence and SNR. The PDF was pre-computed by a Monte-Carlo (MC) simulation based on the mathematical model of JM-OCT before the measurement. A comparison between this new MAP estimator, our previous MAP estimator [2], and the standard mean estimator is presented. The comparisons are performed both by numerical simulation and in vivo measurements of anterior and

  13. Flow cytometric determination of quantitative immunophenotypes

    NASA Astrophysics Data System (ADS)

    Redelman, Douglas; Ensign, Wayne; Roberts, Don

    2001-05-01

    Immunofluorescent flow cytometric analysis of peripheral blood leucocytes is most commonly used to identify and enumerate cells defined by one or more clusters of differentiation (CD) antigens. Although less widely employed, quantitative tests that measure the amounts of CD antigens expressed per cell are used in some situations such as the characterization of lymphomas and leukocytes or the measurement of CD38 on CD3plu8pluT cells in HIV infected individuals. The CD antigens used to identify leukocyte populations are functionally important molecules and it is known that under- or over-expression of some CD antigens can affect cellular responses. For example, high or low expression of CD19 on B cells is associated with autoimmune conditions or depressed antibody responses, respectively. In the current studies, the quantitative expression of CD antigens on T cells, B cells and monocytes was determined in a group of age and sex-matched Marines at several times before and after training exercises. There was substantial variation among these individuals in the quantitative expression of CD antigens and in the number of cells in various populations. However, there was relatively little variation within individuals during the two months they were examined. Thus, the number of cells in leukocyte sub-populations and the amount of CD antigens expressed per cell appear to comprise a characteristic quantitative immunophenotype.

  14. Quantitative determination of engine water ingestion

    NASA Technical Reports Server (NTRS)

    Parikh, P.; Hernan, M.; Sarohia, V.

    1986-01-01

    A nonintrusive optical technique is described for determination of liquid mass flux in a droplet laden airstream. The techniques were developed for quantitative determination of engine water ingestion resulting from heavy rain or wheel spray. Independent measurements of the liquid water content (LWC) of the droplet laden airstream and of the droplet velocities were made at the stimulated nacelle inlet plane for the liquid mass flux determination. The LWC was measured by illuminating and photographing the droplets contained within a thin slice of the flow field by means of a sheet of light from a pulsed laser. A fluorescent dye introduced in the water enchanced the droplet image definition. The droplet velocities were determined from double exposed photographs of the moving droplet field. The technique was initially applied to a steady spray generated in a wind tunnel. It was found that although the spray was initially steady, the aerodynamic breakup process was inherently unsteady. This resulted in a wide variation of the instantaneous LWC of the droplet laden airstream. The standard deviation of ten separate LWC measurements was 31% of the average. However, the liquid mass flux calculated from the average LWC and droplet velocities came within 10% of the known water ingestion rate.

  15. Accurate Determination of Conformational Transitions in Oligomeric Membrane Proteins

    PubMed Central

    Sanz-Hernández, Máximo; Vostrikov, Vitaly V.; Veglia, Gianluigi; De Simone, Alfonso

    2016-01-01

    The structural dynamics governing collective motions in oligomeric membrane proteins play key roles in vital biomolecular processes at cellular membranes. In this study, we present a structural refinement approach that combines solid-state NMR experiments and molecular simulations to accurately describe concerted conformational transitions identifying the overall structural, dynamical, and topological states of oligomeric membrane proteins. The accuracy of the structural ensembles generated with this method is shown to reach the statistical error limit, and is further demonstrated by correctly reproducing orthogonal NMR data. We demonstrate the accuracy of this approach by characterising the pentameric state of phospholamban, a key player in the regulation of calcium uptake in the sarcoplasmic reticulum, and by probing its dynamical activation upon phosphorylation. Our results underline the importance of using an ensemble approach to characterise the conformational transitions that are often responsible for the biological function of oligomeric membrane protein states. PMID:26975211

  16. Fast and accurate determination of modularity and its effect size

    NASA Astrophysics Data System (ADS)

    Treviño, Santiago, III; Nyberg, Amy; Del Genio, Charo I.; Bassler, Kevin E.

    2015-02-01

    We present a fast spectral algorithm for community detection in complex networks. Our method searches for the partition with the maximum value of the modularity via the interplay of several refinement steps that include both agglomeration and division. We validate the accuracy of the algorithm by applying it to several real-world benchmark networks. On all these, our algorithm performs as well or better than any other known polynomial scheme. This allows us to extensively study the modularity distribution in ensembles of Erdős-Rényi networks, producing theoretical predictions for means and variances inclusive of finite-size corrections. Our work provides a way to accurately estimate the effect size of modularity, providing a z-score measure of it and enabling a more informative comparison of networks with different numbers of nodes and links.

  17. Apparatus enables accurate determination of alkali oxides in alkali metals

    NASA Technical Reports Server (NTRS)

    Dupraw, W. A.; Gahn, R. F.; Graab, J. W.; Maple, W. E.; Rosenblum, L.

    1966-01-01

    Evacuated apparatus determines the alkali oxide content of an alkali metal by separating the metal from the oxide by amalgamation with mercury. The apparatus prevents oxygen and moisture from inadvertently entering the system during the sampling and analytical procedure.

  18. Accurate Determination of the Volume of an Irregular Helium Balloon

    NASA Astrophysics Data System (ADS)

    Blumenthal, Jack; Bradvica, Rafaela; Karl, Katherine

    2013-02-01

    In a recent paper, Zable described an experiment with a near-spherical balloon filled with impure helium. Measuring the temperature and the pressure inside and outside the balloon, the lift of the balloon, and the mass of the balloon materials, he described how to use the ideal gas laws and Archimedes' principal to compute the average molecular mass and density of the impure helium. This experiment required that the volume of the near-spherical balloon be determined by some approach, such as measuring the girth. The accuracy of the experiment was largely determined by the balloon volume, which had a reported uncertainty of about 4%.

  19. A Global Approach to Accurate and Automatic Quantitative Analysis of NMR Spectra by Complex Least-Squares Curve Fitting

    NASA Astrophysics Data System (ADS)

    Martin, Y. L.

    The performance of quantitative analysis of 1D NMR spectra depends greatly on the choice of the NMR signal model. Complex least-squares analysis is well suited for optimizing the quantitative determination of spectra containing a limited number of signals (<30) obtained under satisfactory conditions of signal-to-noise ratio (>20). From a general point of view it is concluded, on the basis of mathematical considerations and numerical simulations, that, in the absence of truncation of the free-induction decay, complex least-squares curve fitting either in the time or in the frequency domain and linear-prediction methods are in fact nearly equivalent and give identical results. However, in the situation considered, complex least-squares analysis in the frequency domain is more flexible since it enables the quality of convergence to be appraised at every resonance position. An efficient data-processing strategy has been developed which makes use of an approximate conjugate-gradient algorithm. All spectral parameters (frequency, damping factors, amplitudes, phases, initial delay associated with intensity, and phase parameters of a baseline correction) are simultaneously managed in an integrated approach which is fully automatable. The behavior of the error as a function of the signal-to-noise ratio is theoretically estimated, and the influence of apodization is discussed. The least-squares curve fitting is theoretically proved to be the most accurate approach for quantitative analysis of 1D NMR data acquired with reasonable signal-to-noise ratio. The method enables complex spectral residuals to be sorted out. These residuals, which can be cumulated thanks to the possibility of correcting for frequency shifts and phase errors, extract systematic components, such as isotopic satellite lines, and characterize the shape and the intensity of the spectral distortion with respect to the Lorentzian model. This distortion is shown to be nearly independent of the chemical species

  20. Accurate Optical Target Pose Determination for Applications in Aerial Photogrammetry

    NASA Astrophysics Data System (ADS)

    Cucci, D. A.

    2016-06-01

    We propose a new design for an optical coded target based on concentric circles and a position and orientation determination algorithm optimized for high distances compared to the target size. If two ellipses are fitted on the edge pixels corresponding to the outer and inner circles, quasi-analytical methods are known to obtain the coordinates of the projection of the circles center. We show the limits of these methods for quasi-frontal target orientations and in presence of noise and we propose an iterative refinement algorithm based on a geometric invariant. Next, we introduce a closed form, computationally inexpensive, solution to obtain the target position and orientation given the projected circle center and the parameters of the outer circle projection. The viability of the approach is demonstrated based on aerial pictures taken by an UAV from elevations between 10 to 100 m. We obtain a distance RMS below 0.25 % under 50 m and below 1 % under 100 m with a target size of 90 cm, part of which is a deterministic bias introduced by image exposure.

  1. A quantitative method for determining the robustness of complex networks

    NASA Astrophysics Data System (ADS)

    Qin, Jun; Wu, Hongrun; Tong, Xiaonian; Zheng, Bojin

    2013-06-01

    Most current studies estimate the invulnerability of complex networks using a qualitative method that analyzes the decay rate of network performance. This method results in confusion over the invulnerability of various types of complex networks. By normalizing network performance and defining a baseline, this paper defines the invulnerability index as the integral of the normalized network performance curve minus the baseline. This quantitative method seeks to measure network invulnerability under both edge and node attacks and provides a definition on the distinguishment of the robustness and fragility of networks. To demonstrate the proposed method, three small-world networks were selected as test beds. The simulation results indicate that the proposed invulnerability index can effectively and accurately quantify network resilience and can deal with both the node and edge attacks. The index can provide a valuable reference for determining network invulnerability in future research.

  2. An accurate method of extracting fat droplets in liver images for quantitative evaluation

    NASA Astrophysics Data System (ADS)

    Ishikawa, Masahiro; Kobayashi, Naoki; Komagata, Hideki; Shinoda, Kazuma; Yamaguchi, Masahiro; Abe, Tokiya; Hashiguchi, Akinori; Sakamoto, Michiie

    2015-03-01

    The steatosis in liver pathological tissue images is a promising indicator of nonalcoholic fatty liver disease (NAFLD) and the possible risk of hepatocellular carcinoma (HCC). The resulting values are also important for ensuring the automatic and accurate classification of HCC images, because the existence of many fat droplets is likely to create errors in quantifying the morphological features used in the process. In this study we propose a method that can automatically detect, and exclude regions with many fat droplets by using the feature values of colors, shapes and the arrangement of cell nuclei. We implement the method and confirm that it can accurately detect fat droplets and quantify the fat droplet ratio of actual images. This investigation also clarifies the effective characteristics that contribute to accurate detection.

  3. Accurate determination of the vapor pressure of potassium using optical absorption

    NASA Technical Reports Server (NTRS)

    Shirinzadeh, B.; Wang, C. C.

    1983-01-01

    The vapor pressure of potassium has been measured in absorption using a CW tunable laser and calibrated against the accurate radiative lifetime of the 4s-4p doublet of potassium. An accurate value of 20,850 + or - 30 cal/mol for the heat of vaporization (from the liquid phase) at the melting point was determined.

  4. Quantitative Phase Determination by Using a Michelson Interferometer

    ERIC Educational Resources Information Center

    Pomarico, Juan A.; Molina, Pablo F.; D'Angelo, Cristian

    2007-01-01

    The Michelson interferometer is one of the best established tools for quantitative interferometric measurements. It has been, and is still successfully used, not only for scientific purposes, but it is also introduced in undergraduate courses for qualitative demonstrations as well as for quantitative determination of several properties such as…

  5. Quantitative spectroscopy of hot stars: accurate atomic data applied on a large scale as driver of recent breakthroughs

    NASA Astrophysics Data System (ADS)

    Przybilla, Norbert; Schaffenroth, Veronika; Nieva, Maria-Fernanda

    2015-08-01

    OB-type stars present hotbeds for non-LTE physics because of their strong radiation fields that drive the atmospheric plasma out of local thermodynamic equilibrium. We report on recent breakthroughs in the quantitative analysis of the optical and UV-spectra of OB-type stars that were facilitated by application of accurate and precise atomic data on a large scale. An astophysicist's dream has come true, by bringing observed and model spectra into close match over wide parts of the observed wavelength ranges. This facilitates tight observational constraints to be derived from OB-type stars for wide applications in astrophysics. However, despite the progress made, many details of the modelling may be improved further. We discuss atomic data needs in terms of laboratory measurements and also ab-initio calculations. Particular emphasis is given to quantitative spectroscopy in the near-IR, which will be in focus in the era of the upcoming extremely large telescopes.

  6. Restriction Site Tiling Analysis: accurate discovery and quantitative genotyping of genome-wide polymorphisms using nucleotide arrays

    PubMed Central

    2010-01-01

    High-throughput genotype data can be used to identify genes important for local adaptation in wild populations, phenotypes in lab stocks, or disease-related traits in human medicine. Here we advance microarray-based genotyping for population genomics with Restriction Site Tiling Analysis. The approach simultaneously discovers polymorphisms and provides quantitative genotype data at 10,000s of loci. It is highly accurate and free from ascertainment bias. We apply the approach to uncover genomic differentiation in the purple sea urchin. PMID:20403197

  7. Quantitative determination of stratospheric aerosol characteristics

    NASA Technical Reports Server (NTRS)

    Tingey, D. L.; Potter, J. (Principal Investigator)

    1975-01-01

    The author has identified the following significant results. In the S192 data, a peak was apparent in the lower altitudes that was not present in the shorter wavelengths and grew with increasing wavelength beginning with band 7. For ten S192 wavelengths, the relative altitude increment was determined by knowledge of the relative position of the highest point in the scan arc. Using this scheme, results of scaling and inverting data for passes 47 and 61 were put into two models. Each result had three chart representations: (1) limb brightness measurement, (2) attenuation coefficients, and (3) ratio of the aerosol and Rayleigh coefficients to accentuate layers.

  8. Fluorescence polarization immunoassays for rapid, accurate, and sensitive determination of mycotoxins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Analytical methods for the determination of mycotoxins in foods are commonly based on chromatographic techniques (GC, HPLC or LC-MS). Although these methods permit a sensitive and accurate determination of the analyte, they require skilled personnel and are time-consuming, expensive, and unsuitable ...

  9. High Performance Liquid Chromatography of Vitamin A: A Quantitative Determination.

    ERIC Educational Resources Information Center

    Bohman, Ove; And Others

    1982-01-01

    Experimental procedures are provided for the quantitative determination of Vitamin A (retinol) in food products by analytical liquid chromatography. Standard addition and calibration curve extraction methods are outlined. (SK)

  10. Allele-Specific Quantitative PCR for Accurate, Rapid, and Cost-Effective Genotyping.

    PubMed

    Lee, Han B; Schwab, Tanya L; Koleilat, Alaa; Ata, Hirotaka; Daby, Camden L; Cervera, Roberto Lopez; McNulty, Melissa S; Bostwick, Hannah S; Clark, Karl J

    2016-06-01

    Customizable endonucleases such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) enable rapid generation of mutant strains at genomic loci of interest in animal models and cell lines. With the accelerated pace of generating mutant alleles, genotyping has become a rate-limiting step to understanding the effects of genetic perturbation. Unless mutated alleles result in distinct morphological phenotypes, mutant strains need to be genotyped using standard methods in molecular biology. Classic restriction fragment length polymorphism (RFLP) or sequencing is labor-intensive and expensive. Although simpler than RFLP, current versions of allele-specific PCR may still require post-polymerase chain reaction (PCR) handling such as sequencing, or they are more expensive if allele-specific fluorescent probes are used. Commercial genotyping solutions can take weeks from assay design to result, and are often more expensive than assembling reactions in-house. Key components of commercial assay systems are often proprietary, which limits further customization. Therefore, we developed a one-step open-source genotyping method based on quantitative PCR. The allele-specific qPCR (ASQ) does not require post-PCR processing and can genotype germline mutants through either threshold cycle (Ct) or end-point fluorescence reading. ASQ utilizes allele-specific primers, a locus-specific reverse primer, universal fluorescent probes and quenchers, and hot start DNA polymerase. Individual laboratories can further optimize this open-source system as we completely disclose the sequences, reagents, and thermal cycling protocol. We have tested the ASQ protocol to genotype alleles in five different genes. ASQ showed a 98-100% concordance in genotype scoring with RFLP or Sanger sequencing outcomes. ASQ is time-saving because a single qPCR without post-PCR handling suffices to score

  11. Allele-Specific Quantitative PCR for Accurate, Rapid, and Cost-Effective Genotyping.

    PubMed

    Lee, Han B; Schwab, Tanya L; Koleilat, Alaa; Ata, Hirotaka; Daby, Camden L; Cervera, Roberto Lopez; McNulty, Melissa S; Bostwick, Hannah S; Clark, Karl J

    2016-06-01

    Customizable endonucleases such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) enable rapid generation of mutant strains at genomic loci of interest in animal models and cell lines. With the accelerated pace of generating mutant alleles, genotyping has become a rate-limiting step to understanding the effects of genetic perturbation. Unless mutated alleles result in distinct morphological phenotypes, mutant strains need to be genotyped using standard methods in molecular biology. Classic restriction fragment length polymorphism (RFLP) or sequencing is labor-intensive and expensive. Although simpler than RFLP, current versions of allele-specific PCR may still require post-polymerase chain reaction (PCR) handling such as sequencing, or they are more expensive if allele-specific fluorescent probes are used. Commercial genotyping solutions can take weeks from assay design to result, and are often more expensive than assembling reactions in-house. Key components of commercial assay systems are often proprietary, which limits further customization. Therefore, we developed a one-step open-source genotyping method based on quantitative PCR. The allele-specific qPCR (ASQ) does not require post-PCR processing and can genotype germline mutants through either threshold cycle (Ct) or end-point fluorescence reading. ASQ utilizes allele-specific primers, a locus-specific reverse primer, universal fluorescent probes and quenchers, and hot start DNA polymerase. Individual laboratories can further optimize this open-source system as we completely disclose the sequences, reagents, and thermal cycling protocol. We have tested the ASQ protocol to genotype alleles in five different genes. ASQ showed a 98-100% concordance in genotype scoring with RFLP or Sanger sequencing outcomes. ASQ is time-saving because a single qPCR without post-PCR handling suffices to score

  12. Allele-Specific Quantitative PCR for Accurate, Rapid, and Cost-Effective Genotyping

    PubMed Central

    Lee, Han B.; Schwab, Tanya L.; Koleilat, Alaa; Ata, Hirotaka; Daby, Camden L.; Cervera, Roberto Lopez; McNulty, Melissa S.; Bostwick, Hannah S.; Clark, Karl J.

    2016-01-01

    Customizable endonucleases such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) enable rapid generation of mutant strains at genomic loci of interest in animal models and cell lines. With the accelerated pace of generating mutant alleles, genotyping has become a rate-limiting step to understanding the effects of genetic perturbation. Unless mutated alleles result in distinct morphological phenotypes, mutant strains need to be genotyped using standard methods in molecular biology. Classic restriction fragment length polymorphism (RFLP) or sequencing is labor-intensive and expensive. Although simpler than RFLP, current versions of allele-specific PCR may still require post-polymerase chain reaction (PCR) handling such as sequencing, or they are more expensive if allele-specific fluorescent probes are used. Commercial genotyping solutions can take weeks from assay design to result, and are often more expensive than assembling reactions in-house. Key components of commercial assay systems are often proprietary, which limits further customization. Therefore, we developed a one-step open-source genotyping method based on quantitative PCR. The allele-specific qPCR (ASQ) does not require post-PCR processing and can genotype germline mutants through either threshold cycle (Ct) or end-point fluorescence reading. ASQ utilizes allele-specific primers, a locus-specific reverse primer, universal fluorescent probes and quenchers, and hot start DNA polymerase. Individual laboratories can further optimize this open-source system as we completely disclose the sequences, reagents, and thermal cycling protocol. We have tested the ASQ protocol to genotype alleles in five different genes. ASQ showed a 98–100% concordance in genotype scoring with RFLP or Sanger sequencing outcomes. ASQ is time-saving because a single qPCR without post-PCR handling suffices to score

  13. Factors affecting the accurate determination of cerebrovascular blood flow using high-speed droplet imaging

    NASA Astrophysics Data System (ADS)

    Rudin, Stephen; Divani, Afshin; Wakhloo, Ajay K.; Lieber, Baruch B.; Granger, William; Bednarek, Daniel R.; Yang, Chang-Ying J.

    1998-07-01

    Detailed cerebrovascular blood flow can be more accurately determined radiographically from the new droplet tracking method previously introduced by the authors than from standard soluble contrast techniques. For example, arteriovenous malformation (AVM) transit times which are crucial for proper glue embolization treatments, were shown to be about half when using droplets compared to those measured using soluble contrast techniques. In this work, factors such as x-ray pulse duration, frame rate, system spatial resolution (focal spot size), droplet size, droplet and system contrast parameters, and system noise are considered in relation to their affect on the accurate determination of droplet location and velocity.

  14. Aperture taper determination for the half-scale accurate antenna reflector

    NASA Technical Reports Server (NTRS)

    Lambert, Kevin M.

    1990-01-01

    A simulation is described of a proposed microwave reflectance measurement in which the half scale reflector is used in a compact range type of application. The simulation is used to determine an acceptable aperture taper for the reflector which will allow for accurate measurements. Information on the taper is used in the design of a feed for the reflector.

  15. On canonical cylinder sections for accurate determination of contact angle in microgravity

    SciTech Connect

    Concus, P.; Zabihi, F. California Univ., Berkeley, CA . Dept. of Mathematics); Finn, R. . Dept. of Mathematics)

    1992-07-01

    Large shifts of liquid arising from small changes in certain container shapes in zero gravity can be used as a basis for accurately determining contact angle. Canonical'' geometries for this purpose, recently developed mathematically, are investigated here computationally. It is found that the desired nearly- discontinuous'' behavior can be obtained and that the shifts of liquid have sufficient volume to be readily observed.

  16. On canonical cylinder sections for accurate determination of contact angle in microgravity

    SciTech Connect

    Concus, P.; Zabihi, F. |; Finn, R.

    1992-07-01

    Large shifts of liquid arising from small changes in certain container shapes in zero gravity can be used as a basis for accurately determining contact angle. ``Canonical`` geometries for this purpose, recently developed mathematically, are investigated here computationally. It is found that the desired ``nearly- discontinuous`` behavior can be obtained and that the shifts of liquid have sufficient volume to be readily observed.

  17. The determination of accurate dipole polarizabilities alpha and gamma for the noble gases

    NASA Technical Reports Server (NTRS)

    Rice, Julia E.; Taylor, Peter R.; Lee, Timothy J.; Almlof, Jan

    1991-01-01

    Accurate static dipole polarizabilities alpha and gamma of the noble gases He through Xe were determined using wave functions of similar quality for each system. Good agreement with experimental data for the static polarizability gamma was obtained for Ne and Xe, but not for Ar and Kr. Calculations suggest that the experimental values for these latter ions are too low.

  18. Accurate determination of atomic structure of multiwalled carbon nanotubes by nondestructive nanobeam electron diffraction

    SciTech Connect

    Liu Zejian; Zhang Qi; Qin Luchang

    2005-05-09

    We report a method that allows direct, systematic, and accurate determination of the atomic structure of multiwalled carbon nanotubes by analyzing the scattering intensities on the nonequatorial layer lines in the electron diffraction pattern. Complete structure determination of a quadruple-walled carbon nanotube is described as an example, and it was found that the intertubular distance varied from 0.36 nm to 0.5 nm with a mean value of 0.42 nm.

  19. Communication: Accurate determination of side-chain torsion angle χ1 in proteins: Phenylalanine residues

    NASA Astrophysics Data System (ADS)

    Suardíaz, R.; Crespo-Otero, R.; Pérez, C.; Fabián, J. San; de la Vega, J. M. García

    2011-02-01

    Quantitative side-chain torsion angle χ1 determinations of phenylalanine residues in Desulfovibrio vulgaris flavodoxin are carried out using exclusively the correlation between the experimental vicinal coupling constants and theoretically determined Karplus equations. Karplus coefficients for nine vicinal coupling related with the torsion angle χ1 were calculated using the B3LYP functional and basis sets of different size. Optimized χ1 angles are in outstanding agreement with those previously reported by employing x ray and NMR measurements.

  20. Simple, fast, and accurate methodology for quantitative analysis using Fourier transform infrared spectroscopy, with bio-hybrid fuel cell examples.

    PubMed

    Mackie, David M; Jahnke, Justin P; Benyamin, Marcus S; Sumner, James J

    2016-01-01

    The standard methodologies for quantitative analysis (QA) of mixtures using Fourier transform infrared (FTIR) instruments have evolved until they are now more complicated than necessary for many users' purposes. We present a simpler methodology, suitable for widespread adoption of FTIR QA as a standard laboratory technique across disciplines by occasional users.•Algorithm is straightforward and intuitive, yet it is also fast, accurate, and robust.•Relies on component spectra, minimization of errors, and local adaptive mesh refinement.•Tested successfully on real mixtures of up to nine components. We show that our methodology is robust to challenging experimental conditions such as similar substances, component percentages differing by three orders of magnitude, and imperfect (noisy) spectra. As examples, we analyze biological, chemical, and physical aspects of bio-hybrid fuel cells.

  1. Simple, fast, and accurate methodology for quantitative analysis using Fourier transform infrared spectroscopy, with bio-hybrid fuel cell examples

    PubMed Central

    Mackie, David M.; Jahnke, Justin P.; Benyamin, Marcus S.; Sumner, James J.

    2016-01-01

    The standard methodologies for quantitative analysis (QA) of mixtures using Fourier transform infrared (FTIR) instruments have evolved until they are now more complicated than necessary for many users’ purposes. We present a simpler methodology, suitable for widespread adoption of FTIR QA as a standard laboratory technique across disciplines by occasional users.•Algorithm is straightforward and intuitive, yet it is also fast, accurate, and robust.•Relies on component spectra, minimization of errors, and local adaptive mesh refinement.•Tested successfully on real mixtures of up to nine components. We show that our methodology is robust to challenging experimental conditions such as similar substances, component percentages differing by three orders of magnitude, and imperfect (noisy) spectra. As examples, we analyze biological, chemical, and physical aspects of bio-hybrid fuel cells. PMID:26977411

  2. Quantitative determinations of phenol and resorcinol in pharmaceutical dosage forms by high-pressure liquid chromatography.

    PubMed

    Das Gupta, V

    1976-11-01

    The quantitative determinations of phenol in phenolated calamine lotion USP and of phenol and resorcinol in phenol-resorcinol-boric acid solution by high-pressure liquid chromatography are reported. The procedures are simple, rapid (no special preliminary treatment is required), and accurate. There is no interference from other ingredients of the lotion (bentonite magma, calamine, and zinc oxide) or solution (acetone and boric acid).

  3. Preferential access to genetic information from endogenous hominin ancient DNA and accurate quantitative SNP-typing via SPEX

    PubMed Central

    Brotherton, Paul; Sanchez, Juan J.; Cooper, Alan; Endicott, Phillip

    2010-01-01

    The analysis of targeted genetic loci from ancient, forensic and clinical samples is usually built upon polymerase chain reaction (PCR)-generated sequence data. However, many studies have shown that PCR amplification from poor-quality DNA templates can create sequence artefacts at significant levels. With hominin (human and other hominid) samples, the pervasive presence of highly PCR-amplifiable human DNA contaminants in the vast majority of samples can lead to the creation of recombinant hybrids and other non-authentic artefacts. The resulting PCR-generated sequences can then be difficult, if not impossible, to authenticate. In contrast, single primer extension (SPEX)-based approaches can genotype single nucleotide polymorphisms from ancient fragments of DNA as accurately as modern DNA. A single SPEX-type assay can amplify just one of the duplex DNA strands at target loci and generate a multi-fold depth-of-coverage, with non-authentic recombinant hybrids reduced to undetectable levels. Crucially, SPEX-type approaches can preferentially access genetic information from damaged and degraded endogenous ancient DNA templates over modern human DNA contaminants. The development of SPEX-type assays offers the potential for highly accurate, quantitative genotyping from ancient hominin samples. PMID:19864251

  4. A quantitatively accurate theory of stable crack growth in single phase ductile metal alloys under the influence of cyclic loading

    NASA Astrophysics Data System (ADS)

    Huffman, Peter oel

    Although fatigue has been a well studied phenomenon over the past century and a half, there has yet to be found a quantitative link between fatigue crack growth rates and materials properties. This work serves to establish that link, in the case of well behaved, single phase, ductile metals. The primary mechanisms of fatigue crack growth are identified in general terms, followed by a description of the dependence of the stress intensity factor range on those mechanisms. A method is presented for calculating the crack growth rate for an ideal, linear elastic, non-brittle material, which is assumed to be similar to the crack growth rate for a real material at very small crack growth rate values. The threshold stress intensity factor is discussed as a consequence of "crack tip healing". Residual stresses are accounted for in the form of an approximated residual stress intensity factor. The results of these calculations are compared to data available in the literature. It is concluded that this work presents a new way to consider crack growth with respect to cyclic loading which is quantitatively accurate, and introduces a new way to consider fracture mechanics with respect to the relatively small, cyclic loads, normally associated with fatigue.

  5. Accurate and precise determination of isotopic ratios by MC-ICP-MS: a review.

    PubMed

    Yang, Lu

    2009-01-01

    For many decades the accurate and precise determination of isotope ratios has remained a very strong interest to many researchers due to its important applications in earth, environmental, biological, archeological, and medical sciences. Traditionally, thermal ionization mass spectrometry (TIMS) has been the technique of choice for achieving the highest accuracy and precision. However, recent developments in multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS) have brought a new dimension to this field. In addition to its simple and robust sample introduction, high sample throughput, and high mass resolution, the flat-topped peaks generated by this technique provide for accurate and precise determination of isotope ratios with precision reaching 0.001%, comparable to that achieved with TIMS. These features, in combination with the ability of the ICP source to ionize nearly all elements in the periodic table, have resulted in an increased use of MC-ICP-MS for such measurements in various sample matrices. To determine accurate and precise isotope ratios with MC-ICP-MS, utmost care must be exercised during sample preparation, optimization of the instrument, and mass bias corrections. Unfortunately, there are inconsistencies and errors evident in many MC-ICP-MS publications, including errors in mass bias correction models. This review examines "state-of-the-art" methodologies presented in the literature for achievement of precise and accurate determinations of isotope ratios by MC-ICP-MS. Some general rules for such accurate and precise measurements are suggested, and calculations of combined uncertainty of the data using a few common mass bias correction models are outlined.

  6. Accurate determination of absolute carrier-envelope phase dependence using photo-ionization.

    PubMed

    Sayler, A M; Arbeiter, M; Fasold, S; Adolph, D; Möller, M; Hoff, D; Rathje, T; Fetić, B; Milošević, D B; Fennel, T; Paulus, G G

    2015-07-01

    The carrier-envelope phase (CEP) dependence of few-cycle above-threshold ionization (ATI) of Xe is calibrated for use as a reference measurement for determining and controlling the absolute CEP in other interactions. This is achieved by referencing the CEP-dependent ATI measurements of Xe to measurements of atomic H, which are in turn referenced to ab initio calculations for atomic H. This allows for the accurate determination of the absolute CEP dependence of Xe ATI, which enables relatively easy determination of the offset between the relative CEP measured and/or controlled by typical devices and the absolute CEP in the interaction. PMID:26125386

  7. Rapid and accurate determination of the lignin content of lignocellulosic biomass by solid-state NMR.

    PubMed

    Fu, Li; McCallum, Scott A; Miao, Jianjun; Hart, Courtney; Tudryn, Gregory J; Zhang, Fuming; Linhardt, Robert J

    2015-02-01

    Biofuels and biomaterials, produced from lignocellulosic feedstock, require facile access to cellulose and hemicellulose to be competitive with petroleum processing and sugar-based fermentation. Physical-chemical barriers resulting from lignin complicates the hydrolysis biomass into fermentable sugars. Thus, the amount of lignin within a substrate is critical in determining biomass processing. The application of (13)C cross-polarization, magic-angle spinning, and solid-state nuclear magnetic resonance for the direct quantification of lignin content in biomass is examined. Using a standard curve constructed from pristine lignin and cellulose, the lignin content of a biomass sample is accurately determined through direct measurement without chemical or enzymatic pre-treatment.

  8. Accurate determination of aldehydes in amine catalysts or amines by 2,4-dinitrophenylhydrazine derivatization.

    PubMed

    Barman, Bhajendra N

    2014-01-31

    Carbonyl compounds, specifically aldehydes, present in amine catalysts or amines are determined by reversed-phase liquid chromatography using ultraviolet detection of their corresponding 2,4-dinitrophenylhydrazones. The primary focus has been to establish optimum conditions for determining aldehydes accurately because these add exposure concerns when the amine catalysts are used to manufacture polyurethane products. Concentrations of aldehydes determined by this method are found to vary with the pH of the aqueous amine solution and the derivatization time, the latter being problematic when the derivatization reaction proceeds slowly and not to completion in neutral and basic media. Accurate determination of aldehydes in amines through derivatization can be carried out at an effective solution pH of about 2 and with derivatization time of 20min. Hydrochloric acid has been used for neutralization of an amine. For complete derivatization, it is essential to protonate all nitrogen atoms in the amine. An approach for the determination of an adequate amount of acid needed for complete derivatization has been described. Several 0.2M buffer solutions varying in pH from 4 to 8 have also been used to make amine solutions for carrying out derivatization of aldehydes. These solutions have effective pHs of 10 or higher and provide much lower aldehyde concentrations compared to their true values. Mechanisms for the formation of 2,4-dinitrophenylhydrazones in both acidic and basic media are discussed. PMID:24411140

  9. A highly accurate method for determination of dissolved oxygen: gravimetric Winkler method.

    PubMed

    Helm, Irja; Jalukse, Lauri; Leito, Ivo

    2012-09-01

    A high-accuracy Winkler titration method has been developed for determination of dissolved oxygen concentration. Careful analysis of uncertainty sources relevant to the Winkler method was carried out and the method was optimized for minimizing all uncertainty sources as far as practical. The most important improvements were: gravimetric measurement of all solutions, pre-titration to minimize the effect of iodine volatilization, accurate amperometric end point detection and careful accounting for dissolved oxygen in the reagents. As a result, the developed method is possibly the most accurate method of determination of dissolved oxygen available. Depending on measurement conditions and on the dissolved oxygen concentration the combined standard uncertainties of the method are in the range of 0.012-0.018 mg dm(-3) corresponding to the k=2 expanded uncertainty in the range of 0.023-0.035 mg dm(-3) (0.27-0.38%, relative). This development enables more accurate calibration of electrochemical and optical dissolved oxygen sensors for routine analysis than has been possible before.

  10. Accurate, fast and cost-effective diagnostic test for monosomy 1p36 using real-time quantitative PCR.

    PubMed

    Cunha, Pricila da Silva; Pena, Heloisa B; D'Angelo, Carla Sustek; Koiffmann, Celia P; Rosenfeld, Jill A; Shaffer, Lisa G; Stofanko, Martin; Gonçalves-Dornelas, Higgor; Pena, Sérgio Danilo Junho

    2014-01-01

    Monosomy 1p36 is considered the most common subtelomeric deletion syndrome in humans and it accounts for 0.5-0.7% of all the cases of idiopathic intellectual disability. The molecular diagnosis is often made by microarray-based comparative genomic hybridization (aCGH), which has the drawback of being a high-cost technique. However, patients with classic monosomy 1p36 share some typical clinical characteristics that, together with its common prevalence, justify the development of a less expensive, targeted diagnostic method. In this study, we developed a simple, rapid, and inexpensive real-time quantitative PCR (qPCR) assay for targeted diagnosis of monosomy 1p36, easily accessible for low-budget laboratories in developing countries. For this, we have chosen two target genes which are deleted in the majority of patients with monosomy 1p36: PRKCZ and SKI. In total, 39 patients previously diagnosed with monosomy 1p36 by aCGH, fluorescent in situ hybridization (FISH), and/or multiplex ligation-dependent probe amplification (MLPA) all tested positive on our qPCR assay. By simultaneously using these two genes we have been able to detect 1p36 deletions with 100% sensitivity and 100% specificity. We conclude that qPCR of PRKCZ and SKI is a fast and accurate diagnostic test for monosomy 1p36, costing less than 10 US dollars in reagent costs.

  11. Development and evaluation of a liquid chromatography-mass spectrometry method for rapid, accurate quantitation of malondialdehyde in human plasma.

    PubMed

    Sobsey, Constance A; Han, Jun; Lin, Karen; Swardfager, Walter; Levitt, Anthony; Borchers, Christoph H

    2016-09-01

    Malondialdhyde (MDA) is a commonly used marker of lipid peroxidation in oxidative stress. To provide a sensitive analytical method that is compatible with high throughput, we developed a multiple reaction monitoring-mass spectrometry (MRM-MS) approach using 3-nitrophenylhydrazine chemical derivatization, isotope-labeling, and liquid chromatography (LC) with electrospray ionization (ESI)-tandem mass spectrometry assay to accurately quantify MDA in human plasma. A stable isotope-labeled internal standard was used to compensate for ESI matrix effects. The assay is linear (R(2)=0.9999) over a 20,000-fold concentration range with a lower limit of quantitation of 30fmol (on-column). Intra- and inter-run coefficients of variation (CVs) were <2% and ∼10% respectively. The derivative was stable for >36h at 5°C. Standards spiked into plasma had recoveries of 92-98%. When compared to a common LC-UV method, the LC-MS method found near-identical MDA concentrations. A pilot project to quantify MDA in patient plasma samples (n=26) in a study of major depressive disorder with winter-type seasonal pattern (MDD-s) confirmed known associations between MDA concentrations and obesity (p<0.02). The LC-MS method provides high sensitivity and high reproducibility for quantifying MDA in human plasma. The simple sample preparation and rapid analysis time (5x faster than LC-UV) offers high throughput for large-scale clinical applications. PMID:27437618

  12. Accurate, Fast and Cost-Effective Diagnostic Test for Monosomy 1p36 Using Real-Time Quantitative PCR

    PubMed Central

    Cunha, Pricila da Silva; Pena, Heloisa B.; D'Angelo, Carla Sustek; Koiffmann, Celia P.; Rosenfeld, Jill A.; Shaffer, Lisa G.; Stofanko, Martin; Gonçalves-Dornelas, Higgor; Pena, Sérgio Danilo Junho

    2014-01-01

    Monosomy 1p36 is considered the most common subtelomeric deletion syndrome in humans and it accounts for 0.5–0.7% of all the cases of idiopathic intellectual disability. The molecular diagnosis is often made by microarray-based comparative genomic hybridization (aCGH), which has the drawback of being a high-cost technique. However, patients with classic monosomy 1p36 share some typical clinical characteristics that, together with its common prevalence, justify the development of a less expensive, targeted diagnostic method. In this study, we developed a simple, rapid, and inexpensive real-time quantitative PCR (qPCR) assay for targeted diagnosis of monosomy 1p36, easily accessible for low-budget laboratories in developing countries. For this, we have chosen two target genes which are deleted in the majority of patients with monosomy 1p36: PRKCZ and SKI. In total, 39 patients previously diagnosed with monosomy 1p36 by aCGH, fluorescent in situ hybridization (FISH), and/or multiplex ligation-dependent probe amplification (MLPA) all tested positive on our qPCR assay. By simultaneously using these two genes we have been able to detect 1p36 deletions with 100% sensitivity and 100% specificity. We conclude that qPCR of PRKCZ and SKI is a fast and accurate diagnostic test for monosomy 1p36, costing less than 10 US dollars in reagent costs. PMID:24839341

  13. Quantitative determination of benzyl benzoate in benzyl benzoate lotion NF.

    PubMed

    Gupta, V D; Ho, H W

    1976-07-01

    The development of a simple, shorter and more accurate method than the NF method for the determination of benzyl benzoate in Benzyl Benzoate Lotion NF is discussed. Hydrolyzed benzyl benzoate was measured spectrophotometrically. Interference from other ingredients of the lotion, oleic acid and triethanolamine was almost negligible. The proposed method was completed in approximately 15 minutes, as opposed to the two hours required by the NF procedure.

  14. Quantitative determination and validation of avermectin B1a in commercial products using quantitative nuclear magnetic resonance spectroscopy.

    PubMed

    Hou, Zhuoni; Liang, Xianrui; Du, Liping; Su, Feng; Su, Weike

    2014-09-01

    Nuclear magnetic resonance is defined as a quantitative spectroscopic tool that enables a precise determination of the number of substances in liquids as well as in solids. There is few report demonstrating the application of NMR in the quantification of avermectin B1a (AVB1a ); here, a proton nuclear magnetic resonance spectroscopy ((1) H NMR) using benzene [1-methoxy-4-(2-nitroethyl) (PMN)] as an internal standard and deuterochloroform as an NMR solvent was tested for the quantitative determination of AVB1a . The integrated signal of AVB1a at 5.56 ppm and the signal of PMN at 8.14 ppm in the (1) H NMR spectrum were used for quantification purposes. Parameters of specificity, linearity, accuracy, precision, intermediate precision, range, limit of detection (LOD), limit of quantification (LOQ), stability and robustness were validated. The established method was accurate and precise with good recovery (98.86%) and relative standard deviation (RSD) of assay (0.34%) within the linearity of the calibration curve ranging from 5.08 to 13.58 mg/ml (R(2)  = 0.9999). The LOD and LOQ were 0.009 and 0.029 mg/ml, which indicated the excellent sensitivity of the method. The stability of the method was testified by a calculated RSD of 0.11%. The robustness was testified by modification of four different parameters, and the differences among each parameter were all less than 0.1%. Comparing with the assay described by the manufacturer of avermectin tablets, there was no significant difference between the assay obtained by HPLC and quantitative NMR (qNMR), which indicated qNMR was a simple and efficient method for the determination of AVB1a in commercial formulation products. PMID:24943110

  15. Accurate determination of protein methionine oxidation by stable isotope labeling and LC-MS analysis.

    PubMed

    Liu, Hongcheng; Ponniah, Gomathinayagam; Neill, Alyssa; Patel, Rekha; Andrien, Bruce

    2013-12-17

    Methionine (Met) oxidation is a major modification of proteins, which converts Met to Met sulfoxide as the common product. It is challenging to determine the level of Met sulfoxide, because it can be generated during sample preparation and analysis as an artifact. To determine the level of Met sulfoxide in proteins accurately, an isotope labeling and LC-MS peptide mapping method was developed. Met residues in proteins were fully oxidized using hydrogen peroxide enriched with (18)O atoms before sample preparation. Therefore, it was impossible to generate Met sulfoxide as an artifact during sample preparation. The molecular weight difference of 2 Da between Met sulfoxide with the (16)O atom and Met sulfoxide with the (18)O atom was used to differentiate and calculate the level of Met sulfoxide in the sample originally. Using a recombinant monoclonal antibody as a model protein, much lower levels of Met sulfoxide were detected for the two susceptible Met residues with this new method compared to a typical peptide mapping procedure. The results demonstrated efficient elimination of the analytical artifact during LC-MS peptide mapping for the measurement of Met sulfoxide. This method can thus be used when accurate determination of the level of Met sulfoxide is critical.

  16. Accurate Determination of Torsion and Pure Bending Moment for Viscoelastic Measurements

    NASA Astrophysics Data System (ADS)

    Wang, Yun-Che; Ko, Chih-Chin; Shiau, Li-Ming

    Measurements of time-dependent material properties in the context of linear viscoelasticity, at a given frequency and temperature, require accurate determination of both loading and deformation that are subjected to the testing materials. A pendulum-type viscoelastic spectroscopy is developed to experimentally measure loss tangent and the magnitude of dynamic modulus of solid materials. The mechanical system of the device is based on the behavior of the cantilever beam, and torsion and pure bending moment are generated from the interaction between a permanent magnet and the Helmholtz coils. The strength of the magnetic interactions may be determined with a material with known mechanical properties, such as aluminum 6061T4 alloy. The sensitivity of the torque measurement is on the order of one micro N-m level. With the high accurate torque measurement and deformation detection from a laser-based displacement measurement system, viscoelastic properties of materials can be experimentally measured in different frequency regimes. Sinusoidal driving signals are adopted for measuring complex modulus in the sub-resonant regime, and dc bias driving for creep tests in the low frequency limit. At structural resonant frequencies, the full-width-at-half-maximum (FWHM) method or Lorentzian curve fitting method is adopted to extract material properties. The completion of determining material properties in the wide frequency spectrum may help to identify the deformation mechanisms of the material and to create better models for simulation work.

  17. The Development of a Digital Processing System for Accurate Range Determinations. [for Teleoperator Maneuvering Systems

    NASA Technical Reports Server (NTRS)

    Pujol, A., Jr.

    1983-01-01

    The development of an accurate close range (from 0.0 meters to 30.0 meters) radar system for Teleoperator Maneuvering Systems (TMS) is discussed. The system under investigation is a digital processor that converts incoming signals from the radar system into their related frequency spectra. Identification will be attempted by correlating spectral characteristics with accurate range determinataions. The system will utilize an analog to digital converter for sampling and converting the signal from the radar system into 16-bit digital words (two bytes) for RAM storage, data manipulations, and computations. To remove unwanted frequency components the data will be retrieved from RAM and digitally filtered using large scale integration (LSI) circuits. Filtering will be performed by a biquadratic routine within the chip which carries out the required filter algorithm. For conversion to a frequency spectrum the filtered data will be processed by a Fast Fourier Transform chip. Analysis and identification of spectral characteristics for accurate range determinations will be made by microcomputer computations.

  18. A powerful test of independent assortment that determines genome-wide significance quickly and accurately

    PubMed Central

    Stewart, W C L; Hager, V R

    2016-01-01

    In the analysis of DNA sequences on related individuals, most methods strive to incorporate as much information as possible, with little or no attention paid to the issue of statistical significance. For example, a modern workstation can easily handle the computations needed to perform a large-scale genome-wide inheritance-by-descent (IBD) scan, but accurate assessment of the significance of that scan is often hindered by inaccurate approximations and computationally intensive simulation. To address these issues, we developed gLOD—a test of co-segregation that, for large samples, models chromosome-specific IBD statistics as a collection of stationary Gaussian processes. With this simple model, the parametric bootstrap yields an accurate and rapid assessment of significance—the genome-wide corrected P-value. Furthermore, we show that (i) under the null hypothesis, the limiting distribution of the gLOD is the standard Gumbel distribution; (ii) our parametric bootstrap simulator is approximately 40 000 times faster than gene-dropping methods, and it is more powerful than methods that approximate the adjusted P-value; and, (iii) the gLOD has the same statistical power as the widely used maximum Kong and Cox LOD. Thus, our approach gives researchers the ability to determine quickly and accurately the significance of most large-scale IBD scans, which may contain multiple traits, thousands of families and tens of thousands of DNA sequences. PMID:27245422

  19. A powerful test of independent assortment that determines genome-wide significance quickly and accurately.

    PubMed

    Stewart, W C L; Hager, V R

    2016-08-01

    In the analysis of DNA sequences on related individuals, most methods strive to incorporate as much information as possible, with little or no attention paid to the issue of statistical significance. For example, a modern workstation can easily handle the computations needed to perform a large-scale genome-wide inheritance-by-descent (IBD) scan, but accurate assessment of the significance of that scan is often hindered by inaccurate approximations and computationally intensive simulation. To address these issues, we developed gLOD-a test of co-segregation that, for large samples, models chromosome-specific IBD statistics as a collection of stationary Gaussian processes. With this simple model, the parametric bootstrap yields an accurate and rapid assessment of significance-the genome-wide corrected P-value. Furthermore, we show that (i) under the null hypothesis, the limiting distribution of the gLOD is the standard Gumbel distribution; (ii) our parametric bootstrap simulator is approximately 40 000 times faster than gene-dropping methods, and it is more powerful than methods that approximate the adjusted P-value; and, (iii) the gLOD has the same statistical power as the widely used maximum Kong and Cox LOD. Thus, our approach gives researchers the ability to determine quickly and accurately the significance of most large-scale IBD scans, which may contain multiple traits, thousands of families and tens of thousands of DNA sequences.

  20. Accurate bulk density determination of irregularly shaped translucent and opaque aerogels

    NASA Astrophysics Data System (ADS)

    Petkov, M. P.; Jones, S. M.

    2016-05-01

    We present a volumetric method for accurate determination of bulk density of aerogels, calculated from extrapolated weight of the dry pure solid and volume estimates based on the Archimedes' principle of volume displacement, using packed 100 μm-sized monodispersed glass spheres as a "quasi-fluid" media. Hard particle packing theory is invoked to demonstrate the reproducibility of the apparent density of the quasi-fluid. Accuracy rivaling that of the refractive index method is demonstrated for both translucent and opaque aerogels with different absorptive properties, as well as for aerogels with regular and irregular shapes.

  1. Accurate orbit determination strategies for the tracking and data relay satellites

    NASA Technical Reports Server (NTRS)

    Oza, D. H.; Bolvin, D. T.; Lorah, J. M.; Lee, T.; Doll, C. E.

    1995-01-01

    The National Aeronautics and Space Administration (NASA) has developed the Tracking and Data Relay Satellite (TDRS) System (TDRSS) for tracking and communications support of low Earth-orbiting satellites. TDRSS has the operational capability of providing 85% coverage for TDRSS-user spacecraft. TDRSS currently consists of five geosynchronous spacecraft and the White Sands Complex (WSC) at White Sands, New Mexico. The Bilateration Ranging Transponder System (BRTS) provides range and Doppler measurements for each TDRS. The ground-based BRTS transponders are tracked as if they were TDRSS-user spacecraft. Since the positions of the BRTS transponders are known, their radiometric tracking measurements can be used to provide a well-determined ephemeris for the TDRS spacecraft. For high-accuracy orbit determination of a TDRSS user, such as the Ocean Topography Experiment (TOPEX)/Poseidon spacecraft, high-accuracy TDRS orbits are required. This paper reports on successive refinements in improved techniques and procedures leading to more accurate TDRS orbit determination strategies using the Goddard Trajectory Determination System (GTDS). These strategies range from the standard operational solution using only the BRTS tracking measurements to a sophisticated iterative process involving several successive simultaneous solutions for multiple TDRSs and a TDRSS-user spacecraft. Results are presented for GTDS-generated TDRS ephemerides produced in simultaneous solutions with the TOPEX/Poseidon spacecraft. Strategies with different user spacecraft, as well as schemes for recovering accurate TDRS orbits following a TDRS maneuver, are also presented. In addition, a comprehensive assessment and evaluation of alternative strategies for TDRS orbit determination, excluding BRTS tracking measurements, are presented.

  2. Quantitative and cytotoxic activity determinations on Galanthus nivalis subsp. cilicicus.

    PubMed

    Kaya, G I; Gözler, B

    2005-06-01

    Aerial and underground parts of Galanthus nivalis subsp. cilicicus, a wild-growing species in Turkey, were collected during two different vegetation periods in flowering and fruiting seasons. Herba and bulbus Galanthi were prepared from each specimen. With the aim of collecting data for prospective monographs on this drug, contents of humidity, ash, sulphated ash and total alkaloids were determined according to DAB 10. The specimens were also analyzed quantitatively for two of the principal alkaloids of the genus, galanthamine and lycorine, by using a method based on spectrophotometry complemented with TLC. LC50 values were determined for the ethanolic and alkaloidal extracts of each of the specimens using brine shrimp lethality bioassay.

  3. Accurate measurement of circulating mitochondrial DNA content from human blood samples using real-time quantitative PCR.

    PubMed

    Ajaz, Saima; Czajka, Anna; Malik, Afshan

    2015-01-01

    We describe a protocol to accurately measure the amount of human mitochondrial DNA (MtDNA) in peripheral blood samples which can be modified to quantify MtDNA from other body fluids, human cells, and tissues. This protocol is based on the use of real-time quantitative PCR (qPCR) to quantify the amount of MtDNA relative to nuclear DNA (designated the Mt/N ratio). In the last decade, there have been increasing numbers of studies describing altered MtDNA or Mt/N in circulation in common nongenetic diseases where mitochondrial dysfunction may play a role (for review see Malik and Czajka, Mitochondrion 13:481-492, 2013). These studies are distinct from those looking at genetic mitochondrial disease and are attempting to identify acquired changes in circulating MtDNA content as an indicator of mitochondrial function. However, the methodology being used is not always specific and reproducible. As more than 95 % of the human mitochondrial genome is duplicated in the human nuclear genome, it is important to avoid co-amplification of nuclear pseudogenes. Furthermore, template preparation protocols can also affect the results because of the size and structural differences between the mitochondrial and nuclear genomes. Here we describe how to (1) prepare DNA from blood samples; (2) pretreat the DNA to prevent dilution bias; (3) prepare dilution standards for absolute quantification using the unique primers human mitochondrial genome forward primer (hMitoF3) and human mitochondrial genome reverse primer(hMitoR3) for the mitochondrial genome, and human nuclear genome forward primer (hB2MF1) and human nuclear genome reverse primer (hB2MR1) primers for the human nuclear genome; (4) carry out qPCR for either relative or absolute quantification from test samples; (5) analyze qPCR data; and (6) calculate the sample size to adequately power studies. The protocol presented here is suitable for high-throughput use.

  4. Accurate determination of fiber water-retaining capability at process conditions by headspace gas chromatography.

    PubMed

    Zhang, Shu-Xin; Chai, Xin-Sheng; He, Liang

    2016-09-16

    This work reports on a method for the accurate determination of fiber water-retaining capability at process conditions by headspace gas chromatography (HS-GC) method. The method was based the HS-GC measurement of water vapor on a set closed vials containing in a given amount pulp with different amounts of water addition, from under-saturation to over-saturation. By plotting the equilibrated water vapor signal vs. the amount of water added in pulp, two different trend lines can be observed, in which the transition of the lines corresponds to fiber water-retaining capability. The results showed that the HS-GC method has good measurement precision (much better than the reference method) and good accuracy. The present method can be also used for determining pulp fiber water-retaining capability at the process temperatures in both laboratory research and mill applications. PMID:27554029

  5. Rapid and accurate determination of tissue optical properties using least-squares support vector machines

    PubMed Central

    Barman, Ishan; Dingari, Narahara Chari; Rajaram, Narasimhan; Tunnell, James W.; Dasari, Ramachandra R.; Feld, Michael S.

    2011-01-01

    Diffuse reflectance spectroscopy (DRS) has been extensively applied for the characterization of biological tissue, especially for dysplasia and cancer detection, by determination of the tissue optical properties. A major challenge in performing routine clinical diagnosis lies in the extraction of the relevant parameters, especially at high absorption levels typically observed in cancerous tissue. Here, we present a new least-squares support vector machine (LS-SVM) based regression algorithm for rapid and accurate determination of the absorption and scattering properties. Using physical tissue models, we demonstrate that the proposed method can be implemented more than two orders of magnitude faster than the state-of-the-art approaches while providing better prediction accuracy. Our results show that the proposed regression method has great potential for clinical applications including in tissue scanners for cancer margin assessment, where rapid quantification of optical properties is critical to the performance. PMID:21412464

  6. Rapid and accurate determination of tissue optical properties using least-squares support vector machines.

    PubMed

    Barman, Ishan; Dingari, Narahara Chari; Rajaram, Narasimhan; Tunnell, James W; Dasari, Ramachandra R; Feld, Michael S

    2011-01-01

    Diffuse reflectance spectroscopy (DRS) has been extensively applied for the characterization of biological tissue, especially for dysplasia and cancer detection, by determination of the tissue optical properties. A major challenge in performing routine clinical diagnosis lies in the extraction of the relevant parameters, especially at high absorption levels typically observed in cancerous tissue. Here, we present a new least-squares support vector machine (LS-SVM) based regression algorithm for rapid and accurate determination of the absorption and scattering properties. Using physical tissue models, we demonstrate that the proposed method can be implemented more than two orders of magnitude faster than the state-of-the-art approaches while providing better prediction accuracy. Our results show that the proposed regression method has great potential for clinical applications including in tissue scanners for cancer margin assessment, where rapid quantification of optical properties is critical to the performance. PMID:21412464

  7. Rapid and accurate determination of the lignin content of lignocellulosic biomass by solid-state NMR.

    PubMed

    Fu, Li; McCallum, Scott A; Miao, Jianjun; Hart, Courtney; Tudryn, Gregory J; Zhang, Fuming; Linhardt, Robert J

    2015-02-01

    Biofuels and biomaterials, produced from lignocellulosic feedstock, require facile access to cellulose and hemicellulose to be competitive with petroleum processing and sugar-based fermentation. Physical-chemical barriers resulting from lignin complicates the hydrolysis biomass into fermentable sugars. Thus, the amount of lignin within a substrate is critical in determining biomass processing. The application of (13)C cross-polarization, magic-angle spinning, and solid-state nuclear magnetic resonance for the direct quantification of lignin content in biomass is examined. Using a standard curve constructed from pristine lignin and cellulose, the lignin content of a biomass sample is accurately determined through direct measurement without chemical or enzymatic pre-treatment. PMID:25404762

  8. Rapid and accurate determination of the lignin content of lignocellulosic biomass by solid-state NMR

    PubMed Central

    Fu, Li; McCallum, Scott A.; Miao, Jianjun; Hart, Courtney; Tudryn, Gregory J.; Zhang, Fuming; Linhardt, Robert J.

    2014-01-01

    Biofuels and biomaterials, produced from lignocellulosic feedstock, require facile access to cellulose and hemicellulose to be competitive with petroleum processing and sugar-based fermentation. Physical-chemical barriers resulting from lignin complicates the hydrolysis biomass into fermentable sugars. Thus, the amount of lignin within a substrate is critical in determining biomass processing. The application of 13C cross-polarization, magic-angle spinning, and solid-state nuclear magnetic resonance for the direct quantification of lignin content in biomass is examined. Using a standard curve constructed from pristine lignin and cellulose, the lignin content of a biomass sample is accurately determined through direct measurement without chemical or enzymatic pre-treatment. PMID:25404762

  9. Accurate mass determination, quantification and determination of detection limits in liquid chromatography-high-resolution time-of-flight mass spectrometry: challenges and practical solutions.

    PubMed

    Vergeynst, Leendert; Van Langenhove, Herman; Joos, Pieter; Demeestere, Kristof

    2013-07-30

    Uniform guidelines for the data processing and validation of qualitative and quantitative multi-residue analysis using full-spectrum high-resolution mass spectrometry are scarce. Through systematic research, optimal mass accuracy and sensitivity are obtained after refining the post-processing of the HRMS data. For qualitative analysis, transforming the raw profile spectra to centroid spectra is recommended resulting in a 2.3 fold improved precision on the accurate mass determination of spectrum peaks. However, processing centroid data for quantitative purposes could lead to signal interruption when too narrow mass windows are applied for the construction of extracted ion chromatograms. Therefore, peak integration on the raw profile data is recommended. An optimal width of the mass window of 50 ppm, which is a trade-off between sensitivity and selectivity, was obtained for a TOF instrument providing a resolving power of 20,000 at full width at half maximum (FWHM). For the validation of HRMS analytical methods, widespread concepts such as the signal-to-noise ratios for the determination of decision limits and detection capabilities have shown to be not always applicable because in some cases almost no noise can be detected anymore. A statistical methodology providing a reliable alternative is extended and applied. PMID:23856232

  10. Accurate determination of renal function in patients with intestinal urinary diversions

    SciTech Connect

    McDougal, W.S.; Koch, M.O.

    1986-06-01

    The regular determination of renal function is a critical part of the management of patients who have had the urinary tract reconstructed with intestinal segments. These intestinal segments reabsorb urinary solutes and, thereby, complicate the determination of renal function by conventional methods. Urinary clearances of urea, creatinine and inulin were performed in patients with intestinal segments in the urinary tract and controls under varying diuretic conditions. Patients with intestinal diversions also underwent radioisotopic determination of renal function. The urinary clearances of urea, creatinine and inulin are highly dependent on the rate of urine flow in patients with intestinal segments in the urinary tract. Diuresis maximizes the urinary clearances of these solutes by minimizing intestinal reabsorption. Creatinine clearance prediction from the serum creatinine underestimates true glomerular filtration rate. Radioisotopic determination of renal function correlates poorly with true glomerular filtration rate. Only creatinine clearance measured under diuretic conditions correlates well with true renal function. Urine concentrating ability cannot be assessed accurately in patients with intestinal segments in the urinary tract, since osmolality rapidly equilibrates across the segments.

  11. An improved thin film approximation to accurately determine the optical conductivity of graphene from infrared transmittance

    SciTech Connect

    Weber, J. W.; Bol, A. A.; Sanden, M. C. M. van de

    2014-07-07

    This work presents an improved thin film approximation to extract the optical conductivity from infrared transmittance in a simple yet accurate way. This approximation takes into account the incoherent reflections from the backside of the substrate. These reflections are shown to have a significant effect on the extracted optical conductivity and hence on derived parameters as carrier mobility and density. By excluding the backside reflections, the error for these parameters for typical chemical vapor deposited (CVD) graphene on a silicon substrate can be as high as 17% and 45% for the carrier mobility and density, respectively. For the mid- and near-infrared, the approximation can be simplified such that the real part of the optical conductivity is extracted without the need for a parameterization of the optical conductivity. This direct extraction is shown for Fourier transform infrared (FTIR) transmittance measurements of CVD graphene on silicon in the photon energy range of 370–7000 cm{sup −1}. From the real part of the optical conductivity, the carrier density, mobility, and number of graphene layers are determined but also residue, originating from the graphene transfer, is detected. FTIR transmittance analyzed with the improved thin film approximation is shown to be a non-invasive, easy, and accurate measurement and analysis method for assessing the quality of graphene and can be used for other 2-D materials.

  12. Heuristic determination of quantitative data for knowledge acquisition in medicine.

    PubMed

    Giuse, D A; Giuse, N B; Bankowitz, R A; Miller, R A

    1991-06-01

    Knowledge acquisition for medical knowledge bases can be aided by programs that suggest possible values for portions of the data. The paper presents an experiment which was used in designing a heuristic to help the process of knowledge acquisition. The heuristic helps to determine numerical data from stylized literature excerpts in the context of knowledge acquisition for the QMR medical knowledge base. Quantitative suggestions from the heuristics are shown to agree substantially with the data incorporated in the final version of the knowledge base. The experiment shows the potential of knowledge base specific heuristics in simplifying the task of knowledge base creation. PMID:1868695

  13. Determining suitable image resolutions for accurate supervised crop classification using remote sensing data

    NASA Astrophysics Data System (ADS)

    Löw, Fabian; Duveiller, Grégory

    2013-10-01

    Mapping the spatial distribution of crops has become a fundamental input for agricultural production monitoring using remote sensing. However, the multi-temporality that is often necessary to accurately identify crops and to monitor crop growth generally comes at the expense of coarser observation supports, and can lead to increasingly erroneous class allocations caused by mixed pixels. For a given application like crop classification, the spatial resolution requirement (e.g. in terms of a maximum tolerable pixel size) differs considerably over different landscapes. To analyse the spatial resolution requirements for accurate crop identification via image classification, this study builds upon and extends a conceptual framework established in a previous work1. This framework allows defining quantitatively the spatial resolution requirements for crop monitoring based on simulating how agricultural landscapes, and more specifically the fields covered by a crop of interest, are seen by instruments with increasingly coarser resolving power. The concept of crop specific pixel purity, defined as the degree of homogeneity of the signal encoded in a pixel with respect to the target crop type, is used to analyse how mixed the pixels can be (as they become coarser), without undermining their capacity to describe the desired surface properties. In this case, this framework has been steered towards answering the question: "What is the spatial resolution requirement for crop identification via supervised image classification, in particular minimum and coarsest acceptable pixel sizes, and how do these requirements change over different landscapes?" The framework is applied over four contrasting agro-ecological landscapes in Middle Asia. Inputs to the experiment were eight multi-temporal images from the RapidEye sensor, the simulated pixel sizes range from 6.5 m to 396.5 m. Constraining parameters for crop identification were defined by setting thresholds for classification

  14. Application of an Effective Statistical Technique for an Accurate and Powerful Mining of Quantitative Trait Loci for Rice Aroma Trait.

    PubMed

    Golestan Hashemi, Farahnaz Sadat; Rafii, Mohd Y; Ismail, Mohd Razi; Mohamed, Mahmud Tengku Muda; Rahim, Harun A; Latif, Mohammad Abdul; Aslani, Farzad

    2015-01-01

    When a phenotype of interest is associated with an external/internal covariate, covariate inclusion in quantitative trait loci (QTL) analyses can diminish residual variation and subsequently enhance the ability of QTL detection. In the in vitro synthesis of 2-acetyl-1-pyrroline (2AP), the main fragrance compound in rice, the thermal processing during the Maillard-type reaction between proline and carbohydrate reduction produces a roasted, popcorn-like aroma. Hence, for the first time, we included the proline amino acid, an important precursor of 2AP, as a covariate in our QTL mapping analyses to precisely explore the genetic factors affecting natural variation for rice scent. Consequently, two QTLs were traced on chromosomes 4 and 8. They explained from 20% to 49% of the total aroma phenotypic variance. Additionally, by saturating the interval harboring the major QTL using gene-based primers, a putative allele of fgr (major genetic determinant of fragrance) was mapped in the QTL on the 8th chromosome in the interval RM223-SCU015RM (1.63 cM). These loci supported previous studies of different accessions. Such QTLs can be widely used by breeders in crop improvement programs and for further fine mapping. Moreover, no previous studies and findings were found on simultaneous assessment of the relationship among 2AP, proline and fragrance QTLs. Therefore, our findings can help further our understanding of the metabolomic and genetic basis of 2AP biosynthesis in aromatic rice. PMID:26061689

  15. Accurate determination of the orientational distribution of a fluorescent molecule in a phospholipid membrane.

    PubMed

    Timr, Štěpán; Bondar, Alexey; Cwiklik, Lukasz; Štefl, Martin; Hof, Martin; Vazdar, Mario; Lazar, Josef; Jungwirth, Pavel

    2014-01-30

    Orientation of lipophilic dye molecules within a biological membrane can report on the molecular environment, i.e., the physical and chemical properties of the surrounding membrane. This fact, however, remains under-utilized, largely because of our limited quantitative knowledge of molecular orientational distributions and the fact that robust techniques allowing experimental observation of molecular orientations of dyes in biological membranes are only being developed. In order to begin filling this lack of knowledge and to develop appropriate tools, we have investigated the membrane orientational distribution of the 3-hydroxyflavone-based membrane dye F2N12S. Results of our single- and two-photon polarization microscopy observations of linear dichroism of F2N12S-labeled giant unilamellar vesicles are consistent with a Gaussian-like orientational distribution of the transition dipole moment of the dye, with a mean tilt angle of 53.2 ± 0.1° with respect to the bilayer normal and a standard deviation of 13.3 ± 0.6°. Independently, by combining quantum chemical calculations and molecular dynamics simulations, we obtained very similar values; a mean tilt angle of 48 ± 4° and a standard deviation of 13 ± 2°. The good agreement between the experimentally and computationally obtained values cross-validates both approaches and gives confidence to the results obtained. The results open a door to robust quantitative determinations of orientational distributions of fluorescent molecules (ranging from simple synthetic dyes to fluorescent proteins attached to membrane proteins) associated with lipid membranes. Such determinations enable rational development of a novel class of sensitive fluorescent optical probes, reporting on cellular events through changes in linear dichroism.

  16. Accurate determination of plasmid copy number of flow-sorted cells using droplet digital PCR.

    PubMed

    Jahn, Michael; Vorpahl, Carsten; Türkowsky, Dominique; Lindmeyer, Martin; Bühler, Bruno; Harms, Hauke; Müller, Susann

    2014-06-17

    Many biotechnological processes rely on the expression of a plasmid-based target gene. A constant and sufficient number of plasmids per cell is desired for efficient protein production. To date, only a few methods for the determination of plasmid copy number (PCN) are available, and most of them average the PCN of total populations disregarding heterogeneous distributions. Here, we utilize the highly precise quantification of DNA molecules by droplet digital PCR (ddPCR) and combine it with cell sorting using flow cytometry. A duplex PCR assay was set up requiring only 1000 sorted cells for precise determination of PCN. The robustness of this method was proven by thorough optimization of cell sorting, cell disruption, and PCR conditions. When non plasmid-harboring cells of Pseudomonas putida KT2440 were spiked with different dilutions of the expression plasmid pA-EGFP_B, a PCN from 1 to 64 could be accurately detected. As a proof of principle, induced cultures of P. putida KT2440 producing an EGFP-fused model protein by means of the plasmid pA-EGFP_B were investigated by flow cytometry and showed two distinct subpopulations, fluorescent and nonfluorescent cells. These two subpopulations were sorted for PCN determination with ddPCR. A remarkably diverging plasmid distribution was found within the population, with nonfluorescent cells showing a much lower PCN (≤1) than fluorescent cells (PCN of up to 5) under standard conditions.

  17. Efficient construction of robust artificial neural networks for accurate determination of superficial sample optical properties

    PubMed Central

    Chen, Yu-Wen; Tseng, Sheng-Hao

    2015-01-01

    In general, diffuse reflectance spectroscopy (DRS) systems work with photon diffusion models to determine the absorption coefficient μa and reduced scattering coefficient μs' of turbid samples. However, in some DRS measurement scenarios, such as using short source-detector separations to investigate superficial tissues with comparable μa and μs', photon diffusion models might be invalid or might not have analytical solutions. In this study, a systematic workflow of constructing a rapid, accurate photon transport model that is valid at short source-detector separations (SDSs) and at a wide range of sample albedo is revealed. To create such a model, we first employed a GPU (Graphic Processing Unit) based Monte Carlo model to calculate the reflectance at various sample optical property combinations and established a database at high speed. The database was then utilized to train an artificial neural network (ANN) for determining the sample absorption and reduced scattering coefficients from the reflectance measured at several SDSs without applying spectral constraints. The robustness of the produced ANN model was rigorously validated. We evaluated the performance of a successfully trained ANN using tissue simulating phantoms. We also determined the 500-1000 nm absorption and reduced scattering spectra of in-vivo skin using our ANN model and found that the values agree well with those reported in several independent studies. PMID:25798300

  18. Looking at the cooler hosts - accurate metallicity determination of M dwarfs with and without planets

    NASA Astrophysics Data System (ADS)

    Lindgren, Sara; Heiter, Ulrike

    2015-12-01

    M dwarfs constitute 70% of the stars in the local Galaxy and are becoming attractive targets in the search for Earth-sized planets and planets within the habitable zone. With our research we aim to extend the current understanding of planet formation theory and explore the planet - host metallicity correlation for these cooler hosts.Unlike their solar-type counterparts, the metallicity of M dwarfs is difficult to determine. Their low surface temperature results in plenty of diatomic and triatomic molecules in the photospheric layers. Especially in the optical wavelength region these molecules give rise to a forest of millions of weak lines, making accurate spectroscopy nearly impossible. Previous studies of M dwarfs have therefore established different metallicity calibrations using photometric colors or spectral indices. But these methods exclude the possibility of detailed chemical analysis. High-resolution spectrographs operating in the infrared have recently opened up a new window for investigating M dwarfs. In the infrared the number of molecular transitions is greatly reduced, allowing an accurate continuum placement, and a large number of unblended atomic lines are available. This enabled us to use similar methods as is standard for warmer solar-like stars, and determine the overall metallicity through synthetic spectral fitting.In the first part of our work we used high-resolution spectra taken in the J band (1100-1400nm) with the CRIRES spectrograph, VLT, to verify our method internally and externally by analyzing both components in several M+FGK binaries. In the second part of this study we are analyzing 20 single M dwarfs to achieve a good coverage of effective temperature and metallicity, where our sample covers subtypes M0-M6 and estimated metallicites ranging from +0.8 to -0.8 dex. With these data we aim to derive the to-date most accurate relationship between photometric colors and metallicity for M dwarfs. We will present the current status of our

  19. A method for accurate determination of terminal sequences of viral genomic RNA.

    PubMed

    Weng, Z; Xiong, Z

    1995-09-01

    A combination of ligation-anchored PCR and anchored cDNA cloning techniques were used to clone the termini of the saguaro cactus virus (SCV) RNA genome. The terminal sequences of the viral genome were subsequently determined from the clones. The 5' terminus was cloned by ligation-anchored PCR, whereas the 3' terminus was obtained by a technique we term anchored cDNA cloning. In anchored cDNA cloning, an anchor oligonucleotide was prepared by phosphorylation at the 5' end, followed by addition of a dideoxynucleotide at the 3' end to block the free hydroxyl group. The 5' end of the anchor was subsequently ligated to the 3' end of SCV RNA. The anchor-ligated, chimerical viral RNA was then reverse-transcribed into cDNA using a primer complementary to the anchor. The cDNA containing the complete 3'-terminal sequence was converted into ds-cDNA, cloned, and sequenced. Two restriction sites, one within the viral sequence and one within the primer sequence, were used to facilitate cloning. The combination of these techniques proved to be an easy and accurate way to determine the terminal sequences of SCV RNA genome and should be applicable to any other RNA molecules with unknown terminal sequences. PMID:9132274

  20. Optical coherence tomography enables accurate measurement of equine cartilage thickness for determination of speed of sound.

    PubMed

    Puhakka, Pia H; Te Moller, Nikae C R; Tanska, Petri; Saarakkala, Simo; Tiitu, Virpi; Korhonen, Rami K; Brommer, Harold; Virén, Tuomas; Jurvelin, Jukka S; Töyräs, Juha

    2016-08-01

    Background and purpose - Arthroscopic estimation of articular cartilage thickness is important for scoring of lesion severity, and measurement of cartilage speed of sound (SOS)-a sensitive index of changes in cartilage composition. We investigated the accuracy of optical coherence tomography (OCT) in measurements of cartilage thickness and determined SOS by combining OCT thickness and ultrasound (US) time-of-flight (TOF) measurements. Material and methods - Cartilage thickness measurements from OCT and microscopy images of 94 equine osteochondral samples were compared. Then, SOS in cartilage was determined using simultaneous OCT thickness and US TOF measurements. SOS was then compared with the compositional, structural, and mechanical properties of cartilage. Results - Measurements of non-calcified cartilage thickness using OCT and microscopy were significantly correlated (ρ = 0.92; p < 0.001). With calcified cartilage included, the correlation was ρ = 0.85 (p < 0.001). The mean cartilage SOS (1,636 m/s) was in agreement with the literature. However, SOS and the other properties of cartilage lacked any statistically significant correlation. Interpretation - OCT can give an accurate measurement of articular cartilage thickness. Although SOS measurements lacked accuracy in thin equine cartilage, the concept of SOS measurement using OCT appears promising.

  1. Accurate and easy-to-use assessment of contiguous DNA methylation sites based on proportion competitive quantitative-PCR and lateral flow nucleic acid biosensor.

    PubMed

    Xu, Wentao; Cheng, Nan; Huang, Kunlun; Lin, Yuehe; Wang, Chenguang; Xu, Yuancong; Zhu, Longjiao; Du, Dan; Luo, Yunbo

    2016-06-15

    Many types of diagnostic technologies have been reported for DNA methylation, but they require a standard curve for quantification or only show moderate accuracy. Moreover, most technologies have difficulty providing information on the level of methylation at specific contiguous multi-sites, not to mention easy-to-use detection to eliminate labor-intensive procedures. We have addressed these limitations and report here a cascade strategy that combines proportion competitive quantitative PCR (PCQ-PCR) and lateral flow nucleic acid biosensor (LFNAB), resulting in accurate and easy-to-use assessment. The P16 gene with specific multi-methylated sites, a well-studied tumor suppressor gene, was used as the target DNA sequence model. First, PCQ-PCR provided amplification products with an accurate proportion of multi-methylated sites following the principle of proportionality, and double-labeled duplex DNA was synthesized. Then, a LFNAB strategy was further employed for amplified signal detection via immune affinity recognition, and the exact level of site-specific methylation could be determined by the relative intensity of the test line and internal reference line. This combination resulted in all recoveries being greater than 94%, which are pretty satisfactory recoveries in DNA methylation assessment. Moreover, the developed cascades show significantly high usability as a simple, sensitive, and low-cost tool. Therefore, as a universal platform for sensing systems for the detection of contiguous multi-sites of DNA methylation without external standards and expensive instrumentation, this PCQ-PCR-LFNAB cascade method shows great promise for the point-of-care diagnosis of cancer risk and therapeutics.

  2. Mitochondrial DNA as a non-invasive biomarker: Accurate quantification using real time quantitative PCR without co-amplification of pseudogenes and dilution bias

    SciTech Connect

    Malik, Afshan N.; Shahni, Rojeen; Rodriguez-de-Ledesma, Ana; Laftah, Abas; Cunningham, Phil

    2011-08-19

    Highlights: {yields} Mitochondrial dysfunction is central to many diseases of oxidative stress. {yields} 95% of the mitochondrial genome is duplicated in the nuclear genome. {yields} Dilution of untreated genomic DNA leads to dilution bias. {yields} Unique primers and template pretreatment are needed to accurately measure mitochondrial DNA content. -- Abstract: Circulating mitochondrial DNA (MtDNA) is a potential non-invasive biomarker of cellular mitochondrial dysfunction, the latter known to be central to a wide range of human diseases. Changes in MtDNA are usually determined by quantification of MtDNA relative to nuclear DNA (Mt/N) using real time quantitative PCR. We propose that the methodology for measuring Mt/N needs to be improved and we have identified that current methods have at least one of the following three problems: (1) As much of the mitochondrial genome is duplicated in the nuclear genome, many commonly used MtDNA primers co-amplify homologous pseudogenes found in the nuclear genome; (2) use of regions from genes such as {beta}-actin and 18S rRNA which are repetitive and/or highly variable for qPCR of the nuclear genome leads to errors; and (3) the size difference of mitochondrial and nuclear genomes cause a 'dilution bias' when template DNA is diluted. We describe a PCR-based method using unique regions in the human mitochondrial genome not duplicated in the nuclear genome; unique single copy region in the nuclear genome and template treatment to remove dilution bias, to accurately quantify MtDNA from human samples.

  3. Fuzzy Reasoning to More Accurately Determine Void Areas on Optical Micrographs of Composite Structures

    NASA Technical Reports Server (NTRS)

    Dominquez, Jesus A.; Tate, Lanetra C.; Wright, M. Clara; Caraccio, Anne

    2013-01-01

    Accomplishing the best-performing composite matrix (resin) requires that not only the processing method but also the cure cycle generate low-void-content structures. If voids are present, the performance of the composite matrix will be significantly reduced. This is usually noticed by significant reductions in matrix-dominated properties, such as compression and shear strength. Voids in composite materials are areas that are absent of the composite components: matrix and fibers. The characteristics of the voids and their accurate estimation are critical to determine for high performance composite structures. One widely used method of performing void analysis on a composite structure sample is acquiring optical micrographs or Scanning Electron Microscope (SEM) images of lateral sides of the sample and retrieving the void areas within the micrographs/images using an image analysis technique. Segmentation for the retrieval and subsequent computation of void areas within the micrographs/images is challenging as the gray-scaled values of the void areas are close to the gray-scaled values of the matrix leading to the need of manually performing the segmentation based on the histogram of the micrographs/images to retrieve the void areas. The use of an algorithm developed by NASA and based on Fuzzy Reasoning (FR) proved to overcome the difficulty of suitably differentiate void and matrix image areas with similar gray-scaled values leading not only to a more accurate estimation of void areas on composite matrix micrographs but also to a faster void analysis process as the algorithm is fully autonomous.

  4. Fuzzy Reasoning to More Accurately Determine Void Areas on Optical Micrographs of Composite Structures

    NASA Astrophysics Data System (ADS)

    Dominguez, Jesus A.; Tate, Lanetra C.; Wright, M. Clara; Caraccio, Anne

    2013-12-01

    Accomplishing the best-performing composite matrix (resin) requires that not only the processing method but also the cure cycle generate low-void-content structures. If voids are present, the performance of the composite matrix will be significantly reduced. This is usually noticed by significant reductions in matrix-dominated properties, such as compression and shear strength. Voids in composite materials are areas that are absent of the composite components: matrix and fibers. The characteristics of the voids and their accurate estimation are critical to determine for high performance composite structures. One widely used method of performing void analysis on a composite structure sample is acquiring optical micrographs or Scanning Electron Microscope (SEM) images of lateral sides of the sample and retrieving the void areas within the micrographs/images using an image analysis technique. Segmentation for the retrieval and subsequent computation of void areas within the micrographs/images is challenging as the gray-scaled values of the void areas are close to the gray-scaled values of the matrix leading to the need of manually performing the segmentation based on the histogram of the micrographs/images to retrieve the void areas. The use of an algorithm developed by NASA and based on Fuzzy Reasoning (FR) proved to overcome the difficulty of suitably differentiate void and matrix image areas with similar gray-scaled values leading not only to a more accurate estimation of void areas on composite matrix micrographs but also to a faster void analysis process as the algorithm is fully autonomous.

  5. Quantitative determination of the topological propensities of amyloidogenic peptides.

    PubMed

    Shi, Yuan; Stouten, Pieter F W; Pillalamarri, Nirmala; Barile, Lauren; Rosal, Ramon V; Teichberg, Saul; Bu, Zimei; Callaway, David J E

    2006-03-01

    One of the interesting puzzles of amyloid beta-peptide of Alzheimer's disease (Abeta) is that it appears to polymerize into amyloid fibrils in a parallel beta sheet topology, while smaller subsets of the peptide produce anti-parallel beta sheets. In order to target potential weak points of amyloid fibrils in a rational drug design effort, it would be helpful to understand the forces that drive this change. We have designed two peptides CHQKLVFFAEDYNGKDEAFFVLKQHW and CHQKLVFFAEDYNGKHQKLVFFAEDW that join the significant amyloidogenic Abeta (14-23) sequence HQKLVFFAED in parallel and anti-parallel topologies, respectively. (Here, the word "parallel" refers only to residue sequence and not backbone topology). The N-termini of the hairpins were labeled with the fluorescent dye 5-((((2-iodoacetyl)amino)ethyl)amino)naphthalene-1-sulfonic acid (IAEDANS), forming a fluorescence energy transfer donor-acceptor pair with the C-terminus tryptophan. Circular dichroism results show that the anti-parallel hairpin adopts a beta-sheet conformation, while the parallel hairpin is disordered. Fluorescent Resonance Energy Transfer (FRET) results show that the distance between the donor and the acceptor is significantly shorter in the anti-parallel topology than in the parallel topology. The fluorescence intensity of anti-parallel hairpin also displays a linear concentration dependence, indicating that the FRET observed in the anti-parallel hairpin is from intra-molecular interactions. The results thus provide a quantitative estimate of the relative topological propensities of amyloidogenic peptides. Our FRET and CD results show that beta sheets involving the essential Abeta (14-23) fragment, strongly prefer the anti-parallel topology. Moreover, we provide a quantitative estimate of the relative preference for these two topologies. Such analysis can be repeated for larger subsets of Abeta to determine quantitatively the relative degree of preference for parallel/anti-parallel topologies

  6. Mitochondrial DNA as a non-invasive biomarker: accurate quantification using real time quantitative PCR without co-amplification of pseudogenes and dilution bias.

    PubMed

    Malik, Afshan N; Shahni, Rojeen; Rodriguez-de-Ledesma, Ana; Laftah, Abas; Cunningham, Phil

    2011-08-19

    Circulating mitochondrial DNA (MtDNA) is a potential non-invasive biomarker of cellular mitochondrial dysfunction, the latter known to be central to a wide range of human diseases. Changes in MtDNA are usually determined by quantification of MtDNA relative to nuclear DNA (Mt/N) using real time quantitative PCR. We propose that the methodology for measuring Mt/N needs to be improved and we have identified that current methods have at least one of the following three problems: (1) As much of the mitochondrial genome is duplicated in the nuclear genome, many commonly used MtDNA primers co-amplify homologous pseudogenes found in the nuclear genome; (2) use of regions from genes such as β-actin and 18S rRNA which are repetitive and/or highly variable for qPCR of the nuclear genome leads to errors; and (3) the size difference of mitochondrial and nuclear genomes cause a "dilution bias" when template DNA is diluted. We describe a PCR-based method using unique regions in the human mitochondrial genome not duplicated in the nuclear genome; unique single copy region in the nuclear genome and template treatment to remove dilution bias, to accurately quantify MtDNA from human samples.

  7. Detection and quantitation of trace phenolphthalein (in pharmaceutical preparations and in forensic exhibits) by liquid chromatography-tandem mass spectrometry, a sensitive and accurate method.

    PubMed

    Sharma, Kakali; Sharma, Shiba P; Lahiri, Sujit C

    2013-01-01

    Phenolphthalein, an acid-base indicator and laxative, is important as a constituent of widely used weight-reducing multicomponent food formulations. Phenolphthalein is an useful reagent in forensic science for the identification of blood stains of suspected victims and for apprehending erring officials accepting bribes in graft or trap cases. The pink-colored alkaline hand washes originating from the phenolphthalein-smeared notes can easily be determined spectrophotometrically. But in many cases, colored solution turns colorless with time, which renders the genuineness of bribe cases doubtful to the judiciary. No method is known till now for the detection and identification of phenolphthalein in colorless forensic exhibits with positive proof. Liquid chromatography-tandem mass spectrometry had been found to be most sensitive, accurate method capable of detection and quantitation of trace phenolphthalein in commercial formulations and colorless forensic exhibits with positive proof. The detection limit of phenolphthalein was found to be 1.66 pg/L or ng/mL, and the calibration curve shows good linearity (r(2) = 0.9974). PMID:23106487

  8. Accurate Determination of Comet and Asteroid Orbits Leading to Collision With Earth

    NASA Technical Reports Server (NTRS)

    Roithmayr, Carlos M.; Kay-Bunnell, Linda; Mazanek, Daniel D.; Kumar, Renjith R.; Seywald, Hans; Hausman, Matthew A.

    2005-01-01

    Movements of the celestial bodies in our solar system inspired Isaac Newton to work out his profound laws of gravitation and motion; with one or two notable exceptions, all of those objects move as Newton said they would. But normally harmonious orbital motion is accompanied by the risk of collision, which can be cataclysmic. The Earth s moon is thought to have been produced by such an event, and we recently witnessed magnificent bombardments of Jupiter by several pieces of what was once Comet Shoemaker-Levy 9. Other comets or asteroids may have met the Earth with such violence that dinosaurs and other forms of life became extinct; it is this possibility that causes us to ask how the human species might avoid a similar catastrophe, and the answer requires a thorough understanding of orbital motion. The two red square flags with black square centers displayed are internationally recognized as a warning of an impending hurricane. Mariners and coastal residents who know the meaning of this symbol and the signs evident in the sky and ocean can act in advance to try to protect lives and property; someone who is unfamiliar with the warning signs or chooses to ignore them is in much greater jeopardy. Although collisions between Earth and large comets or asteroids occur much less frequently than landfall of a hurricane, it is imperative that we learn to identify the harbingers of such collisions by careful examination of an object s path. An accurate determination of the orbit of a comet or asteroid is necessary in order to know if, when, and where on the Earth s surface a collision will occur. Generally speaking, the longer the warning time, the better the chance of being able to plan and execute action to prevent a collision. The more accurate the determination of an orbit, the less likely such action will be wasted effort or, what is worse, an effort that increases rather than decreases the probability of a collision. Conditions necessary for a collision to occur are

  9. Accurate and precise determination of critical properties from Gibbs ensemble Monte Carlo simulations

    SciTech Connect

    Dinpajooh, Mohammadhasan; Bai, Peng; Allan, Douglas A.; Siepmann, J. Ilja

    2015-09-21

    Since the seminal paper by Panagiotopoulos [Mol. Phys. 61, 813 (1997)], the Gibbs ensemble Monte Carlo (GEMC) method has been the most popular particle-based simulation approach for the computation of vapor–liquid phase equilibria. However, the validity of GEMC simulations in the near-critical region has been questioned because rigorous finite-size scaling approaches cannot be applied to simulations with fluctuating volume. Valleau [Mol. Simul. 29, 627 (2003)] has argued that GEMC simulations would lead to a spurious overestimation of the critical temperature. More recently, Patel et al. [J. Chem. Phys. 134, 024101 (2011)] opined that the use of analytical tail corrections would be problematic in the near-critical region. To address these issues, we perform extensive GEMC simulations for Lennard-Jones particles in the near-critical region varying the system size, the overall system density, and the cutoff distance. For a system with N = 5500 particles, potential truncation at 8σ and analytical tail corrections, an extrapolation of GEMC simulation data at temperatures in the range from 1.27 to 1.305 yields T{sub c} = 1.3128 ± 0.0016, ρ{sub c} = 0.316 ± 0.004, and p{sub c} = 0.1274 ± 0.0013 in excellent agreement with the thermodynamic limit determined by Potoff and Panagiotopoulos [J. Chem. Phys. 109, 10914 (1998)] using grand canonical Monte Carlo simulations and finite-size scaling. Critical properties estimated using GEMC simulations with different overall system densities (0.296 ≤ ρ{sub t} ≤ 0.336) agree to within the statistical uncertainties. For simulations with tail corrections, data obtained using r{sub cut} = 3.5σ yield T{sub c} and p{sub c} that are higher by 0.2% and 1.4% than simulations with r{sub cut} = 5 and 8σ but still with overlapping 95% confidence intervals. In contrast, GEMC simulations with a truncated and shifted potential show that r{sub cut} = 8σ is insufficient to obtain accurate results. Additional GEMC simulations for hard

  10. Accurate and precise determination of critical properties from Gibbs ensemble Monte Carlo simulations

    NASA Astrophysics Data System (ADS)

    Dinpajooh, Mohammadhasan; Bai, Peng; Allan, Douglas A.; Siepmann, J. Ilja

    2015-09-01

    Since the seminal paper by Panagiotopoulos [Mol. Phys. 61, 813 (1997)], the Gibbs ensemble Monte Carlo (GEMC) method has been the most popular particle-based simulation approach for the computation of vapor-liquid phase equilibria. However, the validity of GEMC simulations in the near-critical region has been questioned because rigorous finite-size scaling approaches cannot be applied to simulations with fluctuating volume. Valleau [Mol. Simul. 29, 627 (2003)] has argued that GEMC simulations would lead to a spurious overestimation of the critical temperature. More recently, Patel et al. [J. Chem. Phys. 134, 024101 (2011)] opined that the use of analytical tail corrections would be problematic in the near-critical region. To address these issues, we perform extensive GEMC simulations for Lennard-Jones particles in the near-critical region varying the system size, the overall system density, and the cutoff distance. For a system with N = 5500 particles, potential truncation at 8σ and analytical tail corrections, an extrapolation of GEMC simulation data at temperatures in the range from 1.27 to 1.305 yields Tc = 1.3128 ± 0.0016, ρc = 0.316 ± 0.004, and pc = 0.1274 ± 0.0013 in excellent agreement with the thermodynamic limit determined by Potoff and Panagiotopoulos [J. Chem. Phys. 109, 10914 (1998)] using grand canonical Monte Carlo simulations and finite-size scaling. Critical properties estimated using GEMC simulations with different overall system densities (0.296 ≤ ρt ≤ 0.336) agree to within the statistical uncertainties. For simulations with tail corrections, data obtained using rcut = 3.5σ yield Tc and pc that are higher by 0.2% and 1.4% than simulations with rcut = 5 and 8σ but still with overlapping 95% confidence intervals. In contrast, GEMC simulations with a truncated and shifted potential show that rcut = 8σ is insufficient to obtain accurate results. Additional GEMC simulations for hard-core square-well particles with various ranges of the

  11. How accurately can the peak skin dose in fluoroscopy be determined using indirect dose metrics?

    SciTech Connect

    Jones, A. Kyle; Ensor, Joe E.; Pasciak, Alexander S.

    2014-07-15

    Purpose: Skin dosimetry is important for fluoroscopically-guided interventions, as peak skin doses (PSD) that result in skin reactions can be reached during these procedures. There is no consensus as to whether or not indirect skin dosimetry is sufficiently accurate for fluoroscopically-guided interventions. However, measuring PSD with film is difficult and the decision to do so must be madea priori. The purpose of this study was to assess the accuracy of different types of indirect dose estimates and to determine if PSD can be calculated within ±50% using indirect dose metrics for embolization procedures. Methods: PSD were measured directly using radiochromic film for 41 consecutive embolization procedures at two sites. Indirect dose metrics from the procedures were collected, including reference air kerma. Four different estimates of PSD were calculated from the indirect dose metrics and compared along with reference air kerma to the measured PSD for each case. The four indirect estimates included a standard calculation method, the use of detailed information from the radiation dose structured report, and two simplified calculation methods based on the standard method. Indirect dosimetry results were compared with direct measurements, including an analysis of uncertainty associated with film dosimetry. Factors affecting the accuracy of the different indirect estimates were examined. Results: When using the standard calculation method, calculated PSD were within ±35% for all 41 procedures studied. Calculated PSD were within ±50% for a simplified method using a single source-to-patient distance for all calculations. Reference air kerma was within ±50% for all but one procedure. Cases for which reference air kerma or calculated PSD exhibited large (±35%) differences from the measured PSD were analyzed, and two main causative factors were identified: unusually small or large source-to-patient distances and large contributions to reference air kerma from cone

  12. A miniature shoe-mounted orientation determination system for accurate indoor heading and trajectory tracking.

    PubMed

    Zhang, Shengzhi; Yu, Shuai; Liu, Chaojun; Liu, Sheng

    2016-06-01

    Tracking the position of pedestrian is urgently demanded when the most commonly used GPS (Global Position System) is unavailable. Benefited from the small size, low-power consumption, and relatively high reliability, micro-electro-mechanical system sensors are well suited for GPS-denied indoor pedestrian heading estimation. In this paper, a real-time miniature orientation determination system (MODS) was developed for indoor heading and trajectory tracking based on a novel dual-linear Kalman filter. The proposed filter precludes the impact of geomagnetic distortions on pitch and roll that the heading is subjected to. A robust calibration approach was designed to improve the accuracy of sensors measurements based on a unified sensor model. Online tests were performed on the MODS with an improved turntable. The results demonstrate that the average RMSE (root-mean-square error) of heading estimation is less than 1°. Indoor heading experiments were carried out with the MODS mounted on the shoe of pedestrian. Besides, we integrated the existing MODS into an indoor pedestrian dead reckoning application as an example of its utility in realistic actions. A human attitude-based walking model was developed to calculate the walking distance. Test results indicate that mean percentage error of indoor trajectory tracking achieves 2% of the total walking distance. This paper provides a feasible alternative for accurate indoor heading and trajectory tracking. PMID:27370490

  13. Diagnostic methodology is critical for accurately determining the prevalence of ichthyophonus infections in wild fish populations

    USGS Publications Warehouse

    Kocan, R.; Dolan, H.; Hershberger, P.

    2011-01-01

    Several different techniques have been employed to detect and identify Ichthyophonus spp. in infected fish hosts; these include macroscopic observation, microscopic examination of tissue squashes, histological evaluation, in vitro culture, and molecular techniques. Examination of the peer-reviewed literature revealed that when more than 1 diagnostic method is used, they often result in significantly different results; for example, when in vitro culture was used to identify infected trout in an experimentally exposed population, 98.7% of infected trout were detected, but when standard histology was used to confirm known infected tissues from wild salmon, it detected ~50% of low-intensity infections and ~85% of high-intensity infections. Other studies on different species reported similar differences. When we examined a possible mechanism to explain the disparity between different diagnostic techniques, we observed non-random distribution of the parasite in 3-dimensionally visualized tissue sections from infected hosts, thus providing a possible explanation for the different sensitivities of commonly used diagnostic techniques. Based on experimental evidence and a review of the peer-reviewed literature, we have concluded that in vitro culture is currently the most accurate diagnostic technique for determining infection prevalence of Ichthyophonus, particularly when the exposure history of the population is not known.

  14. A highly accurate method for the determination of mass and center of mass of a spacecraft

    NASA Technical Reports Server (NTRS)

    Chow, E. Y.; Trubert, M. R.; Egwuatu, A.

    1978-01-01

    An extremely accurate method for the measurement of mass and the lateral center of mass of a spacecraft has been developed. The method was needed for the Voyager spacecraft mission requirement which limited the uncertainty in the knowledge of lateral center of mass of the spacecraft system weighing 750 kg to be less than 1.0 mm (0.04 in.). The method consists of using three load cells symmetrically located at 120 deg apart on a turntable with respect to the vertical axis of the spacecraft and making six measurements for each load cell. These six measurements are taken by cyclic rotations of the load cell turntable and of the spacecraft, about the vertical axis of the measurement fixture. This method eliminates all alignment, leveling, and load cell calibration errors for the lateral center of mass determination, and permits a statistical best fit of the measurement data. An associated data reduction computer program called MASCM has been written to implement this method and has been used for the Voyager spacecraft.

  15. The determination of accurate dipole polarizabilities alpha and gamma for the noble gases

    NASA Technical Reports Server (NTRS)

    Rice, Julia E.; Taylor, Peter R.; Lee, Timothy J.; Almloef, Jan

    1989-01-01

    The static dipole polarizabilities alpha and gamma for the noble gases helium through xenon were determined using large flexible one-particle basis sets in conjunction with high-level treatments of electron correlation. The electron correlation methods include single and double excitation coupled-cluster theory (CCSD), an extension of CCSD that includes a perturbational estimate of connected triple excitations, CCSD(T), and second order perturbation theory (MP2). The computed alpha and gamma values are estimated to be accurate to within a few percent. Agreement with experimental data for the static hyperpolarizability gamma is good for neon and xenon, but for argon and krypton the differences are larger than the combined theoretical and experimental uncertainties. Based on our calculations, we suggest that the experimental value of gamma for argon is too low; adjusting this value would bring the experimental value of gamma for krypton into better agreement with our computed result. The MP2 values for the polarizabilities of neon, argon, krypton and zenon are in reasonabe agreement with the CCSD and CCSD(T) values, suggesting that this less expensive method may be useful in studies of polarizabilities for larger systems.

  16. A miniature shoe-mounted orientation determination system for accurate indoor heading and trajectory tracking

    NASA Astrophysics Data System (ADS)

    Zhang, Shengzhi; Yu, Shuai; Liu, Chaojun; Liu, Sheng

    2016-06-01

    Tracking the position of pedestrian is urgently demanded when the most commonly used GPS (Global Position System) is unavailable. Benefited from the small size, low-power consumption, and relatively high reliability, micro-electro-mechanical system sensors are well suited for GPS-denied indoor pedestrian heading estimation. In this paper, a real-time miniature orientation determination system (MODS) was developed for indoor heading and trajectory tracking based on a novel dual-linear Kalman filter. The proposed filter precludes the impact of geomagnetic distortions on pitch and roll that the heading is subjected to. A robust calibration approach was designed to improve the accuracy of sensors measurements based on a unified sensor model. Online tests were performed on the MODS with an improved turntable. The results demonstrate that the average RMSE (root-mean-square error) of heading estimation is less than 1°. Indoor heading experiments were carried out with the MODS mounted on the shoe of pedestrian. Besides, we integrated the existing MODS into an indoor pedestrian dead reckoning application as an example of its utility in realistic actions. A human attitude-based walking model was developed to calculate the walking distance. Test results indicate that mean percentage error of indoor trajectory tracking achieves 2% of the total walking distance. This paper provides a feasible alternative for accurate indoor heading and trajectory tracking.

  17. Significance of accurate diffraction corrections for the second harmonic wave in determining the acoustic nonlinearity parameter

    SciTech Connect

    Jeong, Hyunjo; Zhang, Shuzeng; Li, Xiongbing; Barnard, Dan

    2015-09-15

    The accurate measurement of acoustic nonlinearity parameter β for fluids or solids generally requires making corrections for diffraction effects due to finite size geometry of transmitter and receiver. These effects are well known in linear acoustics, while those for second harmonic waves have not been well addressed and therefore not properly considered in previous studies. In this work, we explicitly define the attenuation and diffraction corrections using the multi-Gaussian beam (MGB) equations which were developed from the quasilinear solutions of the KZK equation. The effects of making these corrections are examined through the simulation of β determination in water. Diffraction corrections are found to have more significant effects than attenuation corrections, and the β values of water can be estimated experimentally with less than 5% errors when the exact second harmonic diffraction corrections are used together with the negligible attenuation correction effects on the basis of linear frequency dependence between attenuation coefficients, α{sub 2} ≃ 2α{sub 1}.

  18. Final Progress Report: Isotope Identification Algorithm for Rapid and Accurate Determination of Radioisotopes Feasibility Study

    SciTech Connect

    Rawool-Sullivan, Mohini; Bounds, John Alan; Brumby, Steven P.; Prasad, Lakshman; Sullivan, John P.

    2012-04-30

    This is the final report of the project titled, 'Isotope Identification Algorithm for Rapid and Accurate Determination of Radioisotopes,' PMIS project number LA10-HUMANID-PD03. The goal of the work was to demonstrate principles of emulating a human analysis approach towards the data collected using radiation isotope identification devices (RIIDs). It summarizes work performed over the FY10 time period. The goal of the work was to demonstrate principles of emulating a human analysis approach towards the data collected using radiation isotope identification devices (RIIDs). Human analysts begin analyzing a spectrum based on features in the spectrum - lines and shapes that are present in a given spectrum. The proposed work was to carry out a feasibility study that will pick out all gamma ray peaks and other features such as Compton edges, bremsstrahlung, presence/absence of shielding and presence of neutrons and escape peaks. Ultimately success of this feasibility study will allow us to collectively explain identified features and form a realistic scenario that produced a given spectrum in the future. We wanted to develop and demonstrate machine learning algorithms that will qualitatively enhance the automated identification capabilities of portable radiological sensors that are currently being used in the field.

  19. Can scintillation detectors with low spectral resolution accurately determine radionuclides content of building materials?

    PubMed

    Kovler, K; Prilutskiy, Z; Antropov, S; Antropova, N; Bozhko, V; Alfassi, Z B; Lavi, N

    2013-07-01

    The current paper makes an attempt to check whether the scintillation NaI(Tl) detectors, in spite of their poor energy resolution, can determine accurately the content of NORM in building materials. The activity concentrations of natural radionuclides were measured using two types of detectors: (a) NaI(Tl) spectrometer equipped with the special software based on the matrix method of least squares, and (b) high-purity germanium spectrometer. Synthetic compositions with activity concentrations varying in a wide range, from 1/5 to 5 times median activity concentrations of the natural radionuclides available in the earth crust and the samples of popular building materials, such as concrete, pumice and gypsum, were tested, while the density of the tested samples changed in a wide range (from 860 up to 2,410 kg/m(3)). The results obtained in the NaI(Tl) system were similar to those obtained with the HPGe spectrometer, mostly within the uncertainty range. This comparison shows that scintillation spectrometers equipped with a special software aimed to compensate for the lower spectral resolution of NaI(Tl) detectors can be successfully used for the radiation control of mass construction products. PMID:23542118

  20. Rapid, accurate, and direct determination of total lycopene content in tomato paste

    NASA Astrophysics Data System (ADS)

    Bicanic, D.; Anese, M.; Luterotti, S.; Dadarlat, D.; Gibkes, J.; Lubbers, M.

    2003-01-01

    Lycopene that imparts red color to the tomato fruit is the most potent antioxidant among carotenes, an important nutrient and also used as a color ingredient in many food formulations. Since cooked and processed foods derived from tomatoes were shown to provide optimal lycopene boost, products such as paste, puree, juice, etc. are nowadays gaining popularity as dietary sources. The analysis of lycopene in tomato paste (partially dehydrated product prepared by vacuum concentrating tomato juice) is carried out using either high pressure liquid chromatography (HPLC), spectrophotometry, or by evaluating the color. The instability of lycopene during processes of extraction, etc., handling, and disposal of organic solvents makes the preparation of a sample for the analysis a delicate task. Despite a recognized need for accurate and rapid assessment of lycopene in tomato products no such method is available at present. The study described here focuses on a direct determination of a total lycopene content in different tomato pastes by means of the laser optothermal window (LOW) method at 502 nm. The concentration of lycopene in tomato paste ranged between 25 and 150 mg per 100 g product; the results are in excellent agreement with those obtained by spectrophotometry. The time needed to complete LOW analysis is very short, so that decomposition of pigment and the formation of artifacts are minimized. Preliminary results indicate a good degree of reproducibility making the LOW method suitable for routine assays of lycopene content in tomato paste.

  1. Ultrasound Dilution: An Accurate Means of Determining Cardiac Output in Children

    PubMed Central

    Crittendon, Ivory; Dreyer, William J.; Decker, Jamie A; Kim, Jeffrey J.

    2011-01-01

    provides a less invasive method than the traditional pulmonary artery thermodilution for accurately determining cardiac output in children. This is the first validation of the COstatus® system in pediatric patients. Further studies are required to establish its accuracy in pediatric patients with cardiac shunts and other hemodynamically unstable conditions. PMID:21499176

  2. A simple, quantitative method using alginate gel to determine rat colonic tumor volume in vivo.

    PubMed

    Irving, Amy A; Young, Lindsay B; Pleiman, Jennifer K; Konrath, Michael J; Marzella, Blake; Nonte, Michael; Cacciatore, Justin; Ford, Madeline R; Clipson, Linda; Amos-Landgraf, James M; Dove, William F

    2014-04-01

    Many studies of the response of colonic tumors to therapeutics use tumor multiplicity as the endpoint to determine the effectiveness of the agent. These studies can be greatly enhanced by accurate measurements of tumor volume. Here we present a quantitative method to easily and accurately determine colonic tumor volume. This approach uses a biocompatible alginate to create a negative mold of a tumor-bearing colon; this mold is then used to make positive casts of dental stone that replicate the shape of each original tumor. The weight of the dental stone cast correlates highly with the weight of the dissected tumors. After refinement of the technique, overall error in tumor volume was 16.9% ± 7.9% and includes error from both the alginate and dental stone procedures. Because this technique is limited to molding of tumors in the colon, we utilized the Apc(Pirc/+) rat, which has a propensity for developing colonic tumors that reflect the location of the majority of human intestinal tumors. We have successfully used the described method to determine tumor volumes ranging from 4 to 196 mm³. Alginate molding combined with dental stone casting is a facile method for determining tumor volume in vivo without costly equipment or knowledge of analytic software. This broadly accessible method creates the opportunity to objectively study colonic tumors over time in living animals in conjunction with other experiments and without transferring animals from the facility where they are maintained.

  3. A Simple, Quantitative Method Using Alginate Gel to Determine Rat Colonic Tumor Volume In Vivo

    PubMed Central

    Irving, Amy A; Young, Lindsay B; Pleiman, Jennifer K; Konrath, Michael J; Marzella, Blake; Nonte, Michael; Cacciatore, Justin; Ford, Madeline R; Clipson, Linda; Amos-Landgraf, James M; Dove, William F

    2014-01-01

    Many studies of the response of colonic tumors to therapeutics use tumor multiplicity as the endpoint to determine the effectiveness of the agent. These studies can be greatly enhanced by accurate measurements of tumor volume. Here we present a quantitative method to easily and accurately determine colonic tumor volume. This approach uses a biocompatible alginate to create a negative mold of a tumor-bearing colon; this mold is then used to make positive casts of dental stone that replicate the shape of each original tumor. The weight of the dental stone cast correlates highly with the weight of the dissected tumors. After refinement of the technique, overall error in tumor volume was 16.9% ± 7.9% and includes error from both the alginate and dental stone procedures. Because this technique is limited to molding of tumors in the colon, we utilized the ApcPirc/+ rat, which has a propensity for developing colonic tumors that reflect the location of the majority of human intestinal tumors. We have successfully used the described method to determine tumor volumes ranging from 4 to 196 mm3. Alginate molding combined with dental stone casting is a facile method for determining tumor volume in vivo without costly equipment or knowledge of analytic software. This broadly accessible method creates the opportunity to objectively study colonic tumors over time in living animals in conjunction with other experiments and without transferring animals from the facility where they are maintained. PMID:24674588

  4. Quantitative determination of target gene with electrical sensor

    PubMed Central

    Zhang, Xuzhi; Li, Qiufen; Jin, Xianshi; Jiang, Cheng; Lu, Yong; Tavallaie, Roya; Gooding, J. Justin

    2015-01-01

    Integrating loop-mediated isothermal amplification (LAMP) with capacitively coupled contactless conductivity detection (C4D), we have developed an electrical sensor for the simultaneous amplification and detection of specific sequence DNA. Using the O26-wzy gene as a model, the amount of initial target gene could be determined via the threshold time obtained by monitoring the progression of the LAMP reaction in real time. Using the optimal conditions, a detection limit of 12.5 copy/μL can be obtained within 30 min. Monitoring the LAMP reaction by C4D has not only all the advantages that existing electrochemical methods have, but also additional attractive features including being completely free of carryover contamination risk, high simplicity and extremely low cost. These benefits all arise from the fact that the electrodes are separated from the reaction solution, that is C4D is a contactless method. Hence in proof of principle, the new strategy promises a robust, simple, cost-effective and sensitive method for quantitative determination of a target gene, that is applicable either to specialized labs or at point-of-care. PMID:26205714

  5. Quantitative determination of target gene with electrical sensor

    NASA Astrophysics Data System (ADS)

    Zhang, Xuzhi; Li, Qiufen; Jin, Xianshi; Jiang, Cheng; Lu, Yong; Tavallaie, Roya; Gooding, J. Justin

    2015-07-01

    Integrating loop-mediated isothermal amplification (LAMP) with capacitively coupled contactless conductivity detection (C4D), we have developed an electrical sensor for the simultaneous amplification and detection of specific sequence DNA. Using the O26-wzy gene as a model, the amount of initial target gene could be determined via the threshold time obtained by monitoring the progression of the LAMP reaction in real time. Using the optimal conditions, a detection limit of 12.5 copy/μL can be obtained within 30 min. Monitoring the LAMP reaction by C4D has not only all the advantages that existing electrochemical methods have, but also additional attractive features including being completely free of carryover contamination risk, high simplicity and extremely low cost. These benefits all arise from the fact that the electrodes are separated from the reaction solution, that is C4D is a contactless method. Hence in proof of principle, the new strategy promises a robust, simple, cost-effective and sensitive method for quantitative determination of a target gene, that is applicable either to specialized labs or at point-of-care.

  6. Quantitative determination of target gene with electrical sensor.

    PubMed

    Zhang, Xuzhi; Li, Qiufen; Jin, Xianshi; Jiang, Cheng; Lu, Yong; Tavallaie, Roya; Gooding, J Justin

    2015-01-01

    Integrating loop-mediated isothermal amplification (LAMP) with capacitively coupled contactless conductivity detection (C(4)D), we have developed an electrical sensor for the simultaneous amplification and detection of specific sequence DNA. Using the O26-wzy gene as a model, the amount of initial target gene could be determined via the threshold time obtained by monitoring the progression of the LAMP reaction in real time. Using the optimal conditions, a detection limit of 12.5 copy/μL can be obtained within 30 min. Monitoring the LAMP reaction by C(4)D has not only all the advantages that existing electrochemical methods have, but also additional attractive features including being completely free of carryover contamination risk, high simplicity and extremely low cost. These benefits all arise from the fact that the electrodes are separated from the reaction solution, that is C(4)D is a contactless method. Hence in proof of principle, the new strategy promises a robust, simple, cost-effective and sensitive method for quantitative determination of a target gene, that is applicable either to specialized labs or at point-of-care.

  7. [Quantitative determination of morphine in opium powder by addition and correlation method using capillary electrophoresis].

    PubMed

    Sun, Guo-xiang; Miao, Ju-ru; Wang, Yu; Sun, Yu-qing

    2002-01-01

    The morphine in opium powder has been quantitatively determined by addition and correlation method (ACM), in which capillary zone electrophoresis was applied, and the average recovery was 100.6%. The relative standard deviation (RSD) of migration time was not more than 2.4%, the RSD of relative migration time was not more than 1.1%, and the RSD of the relative area was not more than 0.51%. Meanwhile, the contrast test has been done by the calibration curve method with an internal standard correlation. The content of morphine in opium powder determined by ACM was the same as that by using the calibration curve method with an internal standard correlated. The study shows that ACM is simple, quick and accurate.

  8. Wavelet prism decomposition analysis applied to CARS spectroscopy: a tool for accurate and quantitative extraction of resonant vibrational responses.

    PubMed

    Kan, Yelena; Lensu, Lasse; Hehl, Gregor; Volkmer, Andreas; Vartiainen, Erik M

    2016-05-30

    We propose an approach, based on wavelet prism decomposition analysis, for correcting experimental artefacts in a coherent anti-Stokes Raman scattering (CARS) spectrum. This method allows estimating and eliminating a slowly varying modulation error function in the measured normalized CARS spectrum and yields a corrected CARS line-shape. The main advantage of the approach is that the spectral phase and amplitude corrections are avoided in the retrieved Raman line-shape spectrum, thus significantly simplifying the quantitative reconstruction of the sample's Raman response from a normalized CARS spectrum in the presence of experimental artefacts. Moreover, the approach obviates the need for assumptions about the modulation error distribution and the chemical composition of the specimens under study. The method is quantitatively validated on normalized CARS spectra recorded for equimolar aqueous solutions of D-fructose, D-glucose, and their disaccharide combination sucrose. PMID:27410113

  9. Wavelet prism decomposition analysis applied to CARS spectroscopy: a tool for accurate and quantitative extraction of resonant vibrational responses.

    PubMed

    Kan, Yelena; Lensu, Lasse; Hehl, Gregor; Volkmer, Andreas; Vartiainen, Erik M

    2016-05-30

    We propose an approach, based on wavelet prism decomposition analysis, for correcting experimental artefacts in a coherent anti-Stokes Raman scattering (CARS) spectrum. This method allows estimating and eliminating a slowly varying modulation error function in the measured normalized CARS spectrum and yields a corrected CARS line-shape. The main advantage of the approach is that the spectral phase and amplitude corrections are avoided in the retrieved Raman line-shape spectrum, thus significantly simplifying the quantitative reconstruction of the sample's Raman response from a normalized CARS spectrum in the presence of experimental artefacts. Moreover, the approach obviates the need for assumptions about the modulation error distribution and the chemical composition of the specimens under study. The method is quantitatively validated on normalized CARS spectra recorded for equimolar aqueous solutions of D-fructose, D-glucose, and their disaccharide combination sucrose.

  10. Quantitative determination of steroids in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus.

    PubMed

    Gao, Yuan; Xu, Hongyu; Lu, Zhenming; Xu, Zhenghong

    2009-11-01

    This study describes the method of quantitative determination of betulin, ergosterol, cholesterol, lanosterol, stigmasterol and sitosterol in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus. A high performance liquid chromatographic (HPLC) method was applied to separate these steroids. The procedure was carried out on a reversed-phase C, column, using a stepwise gradient of water-methanol as mobile phase with the following profile: 0-10 min, 10% water, 90% methanol; 10-40 min, 3% water, 97% methanol. The flow rate was 1.4 mL/min and the detection wavelength was 202 nm. The analysis was completed within 40 min. The results showed that this method has good reproducibility and satisfactory recoveries for the determination of steroids. The relative standard deviations of the peak areas were less than 2.94% (n = 5) for intraday assays. A good linear correlation was obtained in a range of 0.4-4.8 microg. The recoveries of betulin, ergosterol, cholesterol, lanosterol, stigmasterol, and sitosterol were 100.05%-100.72%, 99.31%-101.04%, 97.52%-101.63%, 96.61%-100.08%, 96.21%-100.76% and 100.04%-100.51%, respectively. This method can be applied to evaluate real samples, and it is rapid, accurate and suitable for the quantitative determination of steroids in the fruiting bodies and submerged-cultured mycelia of Inonotus obliquus.

  11. Determining cleanup levels in bioremediation: Quantitative structure activity relationship techniques

    SciTech Connect

    Arulgnanendran, V.R.J.; Nirmalakhandan, N.

    1995-12-31

    An important feature in the process of planning and initiating bioremediation is the quantification of the toxicity of either an individual chemical or a group of chemicals when multiple chemicals are involved. A laboratory protocol was developed to test the toxicity of single chemicals and mixtures of organic chemicals in a soil medium. Portions of these chemicals are used as a training set to develop Quantitative Structure Activity Relationship (QSAR) models. These predictive models are tested using the chemicals in the testing set, i.e., the remaining chemicals. Moreover mixtures with 10 contaminants in each mixture are tested experimentally to determine joint toxicity for mixtures of chemicals. Using the concepts of Toxic Units, Additivity Index, and Mixture Toxicity Index, the laboratory results are tested for additive, synergistic, or antagonistic effects of the contaminants. These concepts are further validated on mixtures containing eight chemicals that are tested in the laboratory. In addition to the use of the predictive models in evaluating cleanup levels for hazardous waste locations, they are useful to predict microbial toxicity in soils of new chemicals from a congeneric group acting by the same mode of toxicity. These models are applicable when the contaminants act singly or jointly in a mixture.

  12. One-photon mass-analyzed threshold ionization (MATI) spectroscopy of pyridine: determination of accurate ionization energy and cationic structure.

    PubMed

    Lee, Yu Ran; Kang, Do Won; Kim, Hong Lae; Kwon, Chan Ho

    2014-11-01

    Ionization energies and cationic structures of pyridine were intensively investigated utilizing one-photon mass-analyzed threshold ionization (MATI) spectroscopy with vacuum ultraviolet radiation generated by four-wave difference frequency mixing in Kr. The present one-photon high-resolution MATI spectrum of pyridine demonstrated a much finer and richer vibrational structure than that of the previously reported two-photon MATI spectrum. From the MATI spectrum and photoionization efficiency curve, the accurate ionization energy of the ionic ground state of pyridine was confidently determined to be 73,570 ± 6 cm(-1) (9.1215 ± 0.0007 eV). The observed spectrum was almost completely assigned by utilizing Franck-Condon factors and vibrational frequencies calculated through adjustments of the geometrical parameters of cationic pyridine at the B3LYP/cc-pVTZ level. A unique feature unveiled through rigorous analysis was the prominent progression of the 10 vibrational mode, which corresponds to in-plane ring bending, and the combination of other totally symmetric fundamentals with the ring bending overtones, which contribute to the geometrical change upon ionization. Notably, the remaining peaks originate from the upper electronic state ((2)A2), as predicted by high-resolution photoelectron spectroscopy studies and symmetry-adapted cluster configuration interaction calculations. Based on the quantitatively good agreement between the experimental and calculated results, it was concluded that upon ionization the pyridine cation in the ground electronic state should have a planar structure of C(2v) symmetry through the C-N axis.

  13. One-photon mass-analyzed threshold ionization (MATI) spectroscopy of pyridine: Determination of accurate ionization energy and cationic structure

    SciTech Connect

    Lee, Yu Ran; Kang, Do Won; Kim, Hong Lae E-mail: hlkim@kangwon.ac.kr; Kwon, Chan Ho E-mail: hlkim@kangwon.ac.kr

    2014-11-07

    Ionization energies and cationic structures of pyridine were intensively investigated utilizing one-photon mass-analyzed threshold ionization (MATI) spectroscopy with vacuum ultraviolet radiation generated by four-wave difference frequency mixing in Kr. The present one-photon high-resolution MATI spectrum of pyridine demonstrated a much finer and richer vibrational structure than that of the previously reported two-photon MATI spectrum. From the MATI spectrum and photoionization efficiency curve, the accurate ionization energy of the ionic ground state of pyridine was confidently determined to be 73 570 ± 6 cm{sup −1} (9.1215 ± 0.0007 eV). The observed spectrum was almost completely assigned by utilizing Franck-Condon factors and vibrational frequencies calculated through adjustments of the geometrical parameters of cationic pyridine at the B3LYP/cc-pVTZ level. A unique feature unveiled through rigorous analysis was the prominent progression of the 10 vibrational mode, which corresponds to in-plane ring bending, and the combination of other totally symmetric fundamentals with the ring bending overtones, which contribute to the geometrical change upon ionization. Notably, the remaining peaks originate from the upper electronic state ({sup 2}A{sub 2}), as predicted by high-resolution photoelectron spectroscopy studies and symmetry-adapted cluster configuration interaction calculations. Based on the quantitatively good agreement between the experimental and calculated results, it was concluded that upon ionization the pyridine cation in the ground electronic state should have a planar structure of C{sub 2v} symmetry through the C-N axis.

  14. Fluorescent siderophore-based chemosensors: iron(III) quantitative determinations.

    PubMed

    Palanché, T; Marmolle, F; Abdallah, M A; Shanzer, A; Albrecht-Gary, A M

    1999-04-01

    A highly sensitive and selective method is described for a rapid and easy determination of iron(III). This procedure is based on fluorimetric detection combined with the attractive properties of siderophores and biomimetic ligands, which are strong and selective ferric chelators. Azotobactin delta, a bacterial fluorescent siderophore, three fluorescent derivatives of desferriferrioxamine B with a linear structure (NBD-, MA-, NCP-desferriferrioxamine B) and one tripodal biomimetic ligand of desferriferrichrome carrying an anthracenyl fluorescent probe were examined. A very efficient static quenching mechanism by iron was observed for all the ligands considered in this work. Our results identify azotobactin delta as the most promising chemosensor of ferric traces in water, more sensitive than the NBD-desferriferrioxamine B fluorescent ligand. Under more lipophilic conditions, the anthryl-desferriferrichrome biomimetic analogue showed similar analytical potential and was found to be more sensitive than the lipophilic MA- and NCP-desferriferrioxamine B. Their detection limits were respectively 0.5 ng mL-1 for azotobactin delta and 0.6 ng mL-1 for the anthryl tripodal chelator. The calibration curves were linear over the range 0-95 ng mL-1 and 0-180 ng mL-1. Various foreign cations have been examined and only copper(II) and aluminium(III) were shown to interfere when present in similar concentrations as iron(III). The developed procedure using fluorescent siderophores or biomimetic ligands of iron(III) may be applied (1) to monitor iron-(III)-dependent biological systems and (2) to determine iron(III) quantitatively in natural waters and in biological systems.

  15. Modeling of Non-Gravitational Forces for Precise and Accurate Orbit Determination

    NASA Astrophysics Data System (ADS)

    Hackel, Stefan; Gisinger, Christoph; Steigenberger, Peter; Balss, Ulrich; Montenbruck, Oliver; Eineder, Michael

    2014-05-01

    Remote sensing satellites support a broad range of scientific and commercial applications. The two radar imaging satellites TerraSAR-X and TanDEM-X provide spaceborne Synthetic Aperture Radar (SAR) and interferometric SAR data with a very high accuracy. The precise reconstruction of the satellite's trajectory is based on the Global Positioning System (GPS) measurements from a geodetic-grade dual-frequency Integrated Geodetic and Occultation Receiver (IGOR) onboard the spacecraft. The increasing demand for precise radar products relies on validation methods, which require precise and accurate orbit products. An analysis of the orbit quality by means of internal and external validation methods on long and short timescales shows systematics, which reflect deficits in the employed force models. Following the proper analysis of this deficits, possible solution strategies are highlighted in the presentation. The employed Reduced Dynamic Orbit Determination (RDOD) approach utilizes models for gravitational and non-gravitational forces. A detailed satellite macro model is introduced to describe the geometry and the optical surface properties of the satellite. Two major non-gravitational forces are the direct and the indirect Solar Radiation Pressure (SRP). The satellite TerraSAR-X flies on a dusk-dawn orbit with an altitude of approximately 510 km above ground. Due to this constellation, the Sun almost constantly illuminates the satellite, which causes strong across-track accelerations on the plane rectangular to the solar rays. The indirect effect of the solar radiation is called Earth Radiation Pressure (ERP). This force depends on the sunlight, which is reflected by the illuminated Earth surface (visible spectra) and the emission of the Earth body in the infrared spectra. Both components of ERP require Earth models to describe the optical properties of the Earth surface. Therefore, the influence of different Earth models on the orbit quality is assessed. The scope of

  16. Evaluation of Fourier Transform Profilometry for Quantitative Waste Volume Determination under Simulated Hanford Tank Conditions

    SciTech Connect

    Etheridge, J.A.; Jang, P.R.; Leone, T.; Long, Z.; Norton, O.P.; Okhuysen, W.P.; Monts, D.L.; Coggins, T.L.

    2008-07-01

    The Hanford Site is currently in the process of an extensive effort to empty and close its radioactive single-shell and double-shell waste storage tanks. Before this can be accomplished, it is necessary to know how much residual material is left in a given waste tank and the chemical makeup of the residue. The objective of Mississippi State University's Institute for Clean Energy Technology's (ICET) efforts is to develop, fabricate, and deploy inspection tools for the Hanford waste tanks that will (1) be remotely operable; (2) provide quantitative information on the amount of wastes remaining; and (3) provide information on the spatial distribution of chemical and radioactive species of interest. A collaborative arrangement has been established with the Hanford Site to develop probe-based inspection systems for deployment in the waste tanks. ICET is currently developing an in-tank inspection system based on Fourier Transform Profilometry, FTP. FTP is a non-contact, 3-D shape measurement technique. By projecting a fringe pattern onto a target surface and observing its deformation due to surface irregularities from a different view angle, FTP is capable of determining the height (depth) distribution (and hence volume distribution) of the target surface, thus reproducing the profile of the target accurately under a wide variety of conditions. Hence FTP has the potential to be utilized for quantitative determination of residual wastes within Hanford waste tanks. We are conducting a multi-stage performance evaluation of FTP in order to document the accuracy, precision, and operator dependence (minimal) of FTP under conditions similar to those that can be expected to pertain within Hanford waste tanks. The successive stages impose aspects that present increasing difficulty and increasingly more accurate approximations of in-tank environments. In this paper, we report our investigations of the dependence of the analyst upon FTP volume determination results and of the

  17. Plasmid Copy Number Determination by Quantitative Polymerase Chain Reaction.

    PubMed

    Anindyajati; Artarini, A Anita; Riani, Catur; Retnoningrum, Debbie S

    2016-01-01

    Recombinant therapeutic proteins are biopharmaceutical products that develop rapidly for years. Recombinant protein production in certain hosts requires vector expression harboring the gene encoding the corresponding protein. Escherichia coli is the prokaryote organism mostly used in recombinant protein production, commonly using a plasmid as the expression vector. Recombinant protein production is affected by plasmid copy number harboring the encoded gene, hence the determination of plasmid copy number also plays an important role in establishing a recombinant protein production system. On the industrial scale, a low copy number of plasmids are more suitable due to their better stability. In the previous study we constructed pCAD, a plasmid derived from the low copy number pBR322 plasmid. This study was aimed to confirm pCAD's copy number by quantitative polymerase chain reaction (qPCR). Plasmid copy number was determined by comparing the quantification signal from the plasmid to those from the chromosome. Copy number was then calculated by using a known copy number plasmid as a standard. Two pairs of primers, called tdk and ori, were designed for targeting a single gene tdk in the chromosome and a conserved domain in the plasmid's ori, respectively. Primer quality was analyzed in silico using PrimerSelect DNASTAR and PraTo software prior to in vitro evaluation on primer specificity and efficiency as well as optimization of qPCR conditions. Plasmid copy number determination was conducted on E. coli lysates harboring each plasmid, with the number of cells ranging from 10(2)-10(5) cells/μL. Cells were lysed by incubation at 95ºC for 10 minutes, followed by immediate freezing at -4°C. pBR322 plasmid with the copy number of ~19 copies/cell was used as the standard, while pJExpress414-sod plasmid possessing the high copy number pUC ori was also determined to test the method being used. In silico analysis based on primer-primer and primer-template interactions showed

  18. Technique for determination of accurate heat capacities of volatile, powdered, or air-sensitive samples using relaxation calorimetry

    NASA Astrophysics Data System (ADS)

    Marriott, Robert A.; Stancescu, Maria; Kennedy, Catherine A.; White, Mary Anne

    2006-09-01

    We introduce a four-step technique for the accurate determination of the heat capacity of volatile or air-sensitive samples using relaxation calorimetry. The samples are encapsulated in a hermetically sealed differential scanning calorimetry pan, in which there is an internal layer of Apiezon N grease to assist thermal relaxation. Using the Quantum Design physical property measurement system to investigate benzoic acid and copper standards, we find that this method can lead to heat capacity determinations accurate to ±2% over the temperature range of 1-300K, even for very small samples (e.g., <10mg and contributing ca. 20% to the total heat capacity).

  19. Infectious titres of sheep scrapie and bovine spongiform encephalopathy agents cannot be accurately predicted from quantitative laboratory test results.

    PubMed

    González, Lorenzo; Thorne, Leigh; Jeffrey, Martin; Martin, Stuart; Spiropoulos, John; Beck, Katy E; Lockey, Richard W; Vickery, Christopher M; Holder, Thomas; Terry, Linda

    2012-11-01

    It is widely accepted that abnormal forms of the prion protein (PrP) are the best surrogate marker for the infectious agent of prion diseases and, in practice, the detection of such disease-associated (PrP(d)) and/or protease-resistant (PrP(res)) forms of PrP is the cornerstone of diagnosis and surveillance of the transmissible spongiform encephalopathies (TSEs). Nevertheless, some studies question the consistent association between infectivity and abnormal PrP detection. To address this discrepancy, 11 brain samples of sheep affected with natural scrapie or experimental bovine spongiform encephalopathy were selected on the basis of the magnitude and predominant types of PrP(d) accumulation, as shown by immunohistochemical (IHC) examination; contra-lateral hemi-brain samples were inoculated at three different dilutions into transgenic mice overexpressing ovine PrP and were also subjected to quantitative analysis by three biochemical tests (BCTs). Six samples gave 'low' infectious titres (10⁶·⁵ to 10⁶·⁷ LD₅₀ g⁻¹) and five gave 'high titres' (10⁸·¹ to ≥ 10⁸·⁷ LD₅₀ g⁻¹) and, with the exception of the Western blot analysis, those two groups tended to correspond with samples with lower PrP(d)/PrP(res) results by IHC/BCTs. However, no statistical association could be confirmed due to high individual sample variability. It is concluded that although detection of abnormal forms of PrP by laboratory methods remains useful to confirm TSE infection, infectivity titres cannot be predicted from quantitative test results, at least for the TSE sources and host PRNP genotypes used in this study. Furthermore, the near inverse correlation between infectious titres and Western blot results (high protease pre-treatment) argues for a dissociation between infectivity and PrP(res).

  20. Quantitative determinations of antipyrine and benzocaine in ear drops by high-pressure liquid chromatography.

    PubMed

    Gupta, V D; Sachanandani, S

    1977-06-01

    Antipyrine and benzocaine were determined quantitatively in ear drops by high-pressure liquid chromatography on an octadecyltrichlorosilane permanently bonded to a Si-C column, using 0.02 M KH2PO4 in methanol-water as the mobile phase. Both compounds can be assayed in combination directly without interference from each other or from oxyquinoline sulfate (the preservative). The method is accurate, precise (estimated universe standard deviations of 0.68% for antipyrine and 1.18% for benzocaine), simple, and short (requires 30 min versus 2-3 hr by the NF method). The method was tried on a commercial product with excellent results and can be used in the presence of decomposition products.

  1. Accurate determination of specific heat at high temperatures using the flash diffusivity method

    NASA Technical Reports Server (NTRS)

    Vandersande, J. W.; Zoltan, A.; Wood, C.

    1989-01-01

    The flash diffusivity method of Parker et al. (1961) was used to measure accurately the specific heat of test samples simultaneously with thermal diffusivity, thus obtaining the thermal conductivity of these materials directly. The accuracy of data obtained on two types of materials (n-type silicon-germanium alloys and niobium), was + or - 3 percent. It is shown that the method is applicable up to at least 1300 K.

  2. Accurate determination of solid and liquid dispersions from spectra channeled with the Fabry-Perot interferometer.

    PubMed

    Khashan, M A; Nassif, A Y

    1997-09-20

    The band spacing of the fringes of equal chromatic order of a thin Fabry-Perot interferometer is compared when this interferometer contains air, a solid, or a liquid. This comparison enables the dispersion of the group velocity of light in these media to be known accurately to 2.4 parts in one thousand. The Sellmeier dispersion function is used to deduce the refractive indices with the same degree of accuracy. The order-transformation method is used to find the exact order values from the roughly known optical thickness. The exact order values for air and the sample are used to find the refractive index accurately to approximately 3 x 10(-5). A least-squares fitting of the accurate experimental data to the Sellmeier dispersion function enables the coefficients of the latter to be more precisely defined for solids such as glass and mica and for liquids such as glycerin and distilled water. The atomic parameters such as the density of states and the absorption wavelengths in the ultraviolet region of the spectrum for the given samples are deduced from the more precisely found Sellmeier coefficients. PMID:18259554

  3. Validation of Reference Genes for Accurate Normalization of Gene Expression in Lilium davidii var. unicolor for Real Time Quantitative PCR

    PubMed Central

    Zhang, Jing; Teixeira da Silva, Jaime A.; Wang, ChunXia; Sun, HongMei

    2015-01-01

    Lilium is an important commercial market flower bulb. qRT-PCR is an extremely important technique to track gene expression levels. The requirement of suitable reference genes for normalization has become increasingly significant and exigent. The expression of internal control genes in living organisms varies considerably under different experimental conditions. For economically important Lilium, only a limited number of reference genes applied in qRT-PCR have been reported to date. In this study, the expression stability of 12 candidate genes including α-TUB, β-TUB, ACT, eIF, GAPDH, UBQ, UBC, 18S, 60S, AP4, FP, and RH2, in a diverse set of 29 samples representing different developmental processes, three stress treatments (cold, heat, and salt) and different organs, has been evaluated. For different organs, the combination of ACT, GAPDH, and UBQ is appropriate whereas ACT together with AP4, or ACT along with GAPDH is suitable for normalization of leaves and scales at different developmental stages, respectively. In leaves, scales and roots under stress treatments, FP, ACT and AP4, respectively showed the most stable expression. This study provides a guide for the selection of a reference gene under different experimental conditions, and will benefit future research on more accurate gene expression studies in a wide variety of Lilium genotypes. PMID:26509446

  4. Validation of Reference Genes for Accurate Normalization of Gene Expression in Lilium davidii var. unicolor for Real Time Quantitative PCR.

    PubMed

    Li, XueYan; Cheng, JinYun; Zhang, Jing; Teixeira da Silva, Jaime A; Wang, ChunXia; Sun, HongMei

    2015-01-01

    Lilium is an important commercial market flower bulb. qRT-PCR is an extremely important technique to track gene expression levels. The requirement of suitable reference genes for normalization has become increasingly significant and exigent. The expression of internal control genes in living organisms varies considerably under different experimental conditions. For economically important Lilium, only a limited number of reference genes applied in qRT-PCR have been reported to date. In this study, the expression stability of 12 candidate genes including α-TUB, β-TUB, ACT, eIF, GAPDH, UBQ, UBC, 18S, 60S, AP4, FP, and RH2, in a diverse set of 29 samples representing different developmental processes, three stress treatments (cold, heat, and salt) and different organs, has been evaluated. For different organs, the combination of ACT, GAPDH, and UBQ is appropriate whereas ACT together with AP4, or ACT along with GAPDH is suitable for normalization of leaves and scales at different developmental stages, respectively. In leaves, scales and roots under stress treatments, FP, ACT and AP4, respectively showed the most stable expression. This study provides a guide for the selection of a reference gene under different experimental conditions, and will benefit future research on more accurate gene expression studies in a wide variety of Lilium genotypes. PMID:26509446

  5. Quantitative Fourier transform ion cyclotron resonance mass spectrometry--the determination of creatinine by isotope dilution mass spectrometry.

    PubMed

    Bristow, Tony; Stokes, Peter; O'Connor, Gavin

    2005-01-01

    Accurate quantitation has been demonstrated on many different types of mass spectrometer. However, quantitative applications of Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) have been limited. In this study, the quantitative potential of FTICRMS has been investigated using an exact matching isotope dilution method for the determination of creatinine in serum. Creatinine is an important clinical biomarker and its measurement is used as an assessment of renal function. The quantitation of creatinine was selected because a high-accuracy high-performance liquid chromatography/mass spectrometry (HPLC/MS) determination using a triple quadrupole mass spectrometer has already been successfully developed in-house. Therefore, a direct comparison of the quantitative capability of FTICRMS could be made against an established method. The accuracy of the quantitation of creatinine was found to be equivalent to that obtained using LC/MS. However, the expanded measurement uncertainty (k = 2) was larger, at 6%, when using FTICRMS compared with 1% when using HPLC/MS with the triple quadrupole mass spectrometer.

  6. Determining Sample Size for Accurate Estimation of the Squared Multiple Correlation Coefficient.

    ERIC Educational Resources Information Center

    Algina, James; Olejnik, Stephen

    2000-01-01

    Discusses determining sample size for estimation of the squared multiple correlation coefficient and presents regression equations that permit determination of the sample size for estimating this parameter for up to 20 predictor variables. (SLD)

  7. A Simple yet Accurate Method for Students to Determine Asteroid Rotation Periods from Fragmented Light Curve Data

    ERIC Educational Resources Information Center

    Beare, R. A.

    2008-01-01

    Professional astronomers use specialized software not normally available to students to determine the rotation periods of asteroids from fragmented light curve data. This paper describes a simple yet accurate method based on Microsoft Excel[R] that enables students to find periods in asteroid light curve and other discontinuous time series data of…

  8. Evaluation of Faecalibacterium 16S rDNA genetic markers for accurate identification of swine faecal waste by quantitative PCR.

    PubMed

    Duan, Chuanren; Cui, Yamin; Zhao, Yi; Zhai, Jun; Zhang, Baoyun; Zhang, Kun; Sun, Da; Chen, Hang

    2016-10-01

    A genetic marker within the 16S rRNA gene of Faecalibacterium was identified for use in a quantitative PCR (qPCR) assay to detect swine faecal contamination in water. A total of 146,038 bacterial sequences were obtained using 454 pyrosequencing. By comparative bioinformatics analysis of Faecalibacterium sequences with those of numerous swine and other animal species, swine-specific Faecalibacterium 16S rRNA gene sequences were identified and Polymerase Chain Okabe (PCR) primer sets designed and tested against faecal DNA samples from swine and non-swine sources. Two PCR primer sets, PFB-1 and PFB-2, showed the highest specificity to swine faecal waste and had no cross-reaction with other animal samples. PFB-1 and PFB-2 amplified 16S rRNA gene sequences from 50 samples of swine with positive ratios of 86 and 90%, respectively. We compared swine-specific Faecalibacterium qPCR assays for the purpose of quantifying the newly identified markers. The quantification limits (LOQs) of PFB-1 and PFB-2 markers in environmental water were 6.5 and 2.9 copies per 100 ml, respectively. Of the swine-associated assays tested, PFB-2 was more sensitive in detecting the swine faecal waste and quantifying the microbial load. Furthermore, the microbial abundance and diversity of the microbiomes of swine and other animal faeces were estimated using operational taxonomic units (OTUs). The species specificity was demonstrated for the microbial populations present in various animal faeces. PMID:27353369

  9. New methods determine pour point more accurately than ASTM D-97

    SciTech Connect

    Khan, H.U.; Dilawar, S.V.K.; Nautiyal, S.P.; Srivastava, S.P. )

    1993-11-01

    A new, alternative method determines petroleum fluid pour points with [+-] 1 C. precision and better accuracy than the standard ASTM D-97 procedure. The new method measures the pour point of transparent fluids by determining wax appearance temperature (WAT). Also, pour points of waxy crude oils can be determined by measuring a flow characteristic called restart pressure.

  10. Evaluation of errors in quantitative determination of asbestos in rock

    NASA Astrophysics Data System (ADS)

    Baietto, Oliviero; Marini, Paola; Vitaliti, Martina

    2016-04-01

    The quantitative determination of the content of asbestos in rock matrices is a complex operation which is susceptible to important errors. The principal methodologies for the analysis are Scanning Electron Microscopy (SEM) and Phase Contrast Optical Microscopy (PCOM). Despite the PCOM resolution is inferior to that of SEM, PCOM analysis has several advantages, including more representativity of the analyzed sample, more effective recognition of chrysotile and a lower cost. The DIATI LAA internal methodology for the analysis in PCOM is based on a mild grinding of a rock sample, its subdivision in 5-6 grain size classes smaller than 2 mm and a subsequent microscopic analysis of a portion of each class. The PCOM is based on the optical properties of asbestos and of the liquids with note refractive index in which the particles in analysis are immersed. The error evaluation in the analysis of rock samples, contrary to the analysis of airborne filters, cannot be based on a statistical distribution. In fact for airborne filters a binomial distribution (Poisson), which theoretically defines the variation in the count of fibers resulting from the observation of analysis fields, chosen randomly on the filter, can be applied. The analysis in rock matrices instead cannot lean on any statistical distribution because the most important object of the analysis is the size of the of asbestiform fibers and bundles of fibers observed and the resulting relationship between the weights of the fibrous component compared to the one granular. The error evaluation generally provided by public and private institutions varies between 50 and 150 percent, but there are not, however, specific studies that discuss the origin of the error or that link it to the asbestos content. Our work aims to provide a reliable estimation of the error in relation to the applied methodologies and to the total content of asbestos, especially for the values close to the legal limits. The error assessments must

  11. Toward Quantitatively Accurate Calculation of the Redox-Associated Acid–Base and Ligand Binding Equilibria of Aquacobalamin

    DOE PAGES

    Johnston, Ryne C.; Zhou, Jing; Smith, Jeremy C.; Parks, Jerry M.

    2016-07-08

    In redox processes in complex transition metal-containing species are often intimately associated with changes in ligand protonation states and metal coordination number. Moreover, a major challenge is therefore to develop consistent computational approaches for computing pH-dependent redox and ligand dissociation properties of organometallic species. Reduction of the Co center in the vitamin B12 derivative aquacobalamin can be accompanied by ligand dissociation, protonation, or both, making these properties difficult to compute accurately. We examine this challenge here by using density functional theory and continuum solvation to compute Co ligand binding equilibrium constants (Kon/off), pKas and reduction potentials for models of aquacobalaminmore » in aqueous solution. We consider two models for cobalamin ligand coordination: the first follows the hexa, penta, tetra coordination scheme for CoIII, CoII, and CoI species, respectively, and the second model features saturation of each vacant axial coordination site on CoII and CoI species with a single, explicit water molecule to maintain six directly interacting ligands or water molecules in each oxidation state. Comparing these two coordination schemes in combination with five dispersion-corrected density functionals, we find that the accuracy of the computed properties is largely independent of the scheme used, but including only a continuum representation of the solvent yields marginally better results than saturating the first solvation shell around Co throughout. PBE performs best, displaying balanced accuracy and superior performance overall, with RMS errors of 80 mV for seven reduction potentials, 2.0 log units for five pKas and 2.3 log units for two log Kon/off values for the aquacobalamin system. Furthermore, we find that the BP86 functional commonly used in corrinoid studies suffers from erratic behavior and inaccurate descriptions of Co axial ligand binding, leading to substantial errors in predicted

  12. Toward Quantitatively Accurate Calculation of the Redox-Associated Acid-Base and Ligand Binding Equilibria of Aquacobalamin.

    PubMed

    Johnston, Ryne C; Zhou, Jing; Smith, Jeremy C; Parks, Jerry M

    2016-08-01

    Redox processes in complex transition metal-containing species are often intimately associated with changes in ligand protonation states and metal coordination number. A major challenge is therefore to develop consistent computational approaches for computing pH-dependent redox and ligand dissociation properties of organometallic species. Reduction of the Co center in the vitamin B12 derivative aquacobalamin can be accompanied by ligand dissociation, protonation, or both, making these properties difficult to compute accurately. We examine this challenge here by using density functional theory and continuum solvation to compute Co-ligand binding equilibrium constants (Kon/off), pKas, and reduction potentials for models of aquacobalamin in aqueous solution. We consider two models for cobalamin ligand coordination: the first follows the hexa, penta, tetra coordination scheme for Co(III), Co(II), and Co(I) species, respectively, and the second model features saturation of each vacant axial coordination site on Co(II) and Co(I) species with a single, explicit water molecule to maintain six directly interacting ligands or water molecules in each oxidation state. Comparing these two coordination schemes in combination with five dispersion-corrected density functionals, we find that the accuracy of the computed properties is largely independent of the scheme used, but including only a continuum representation of the solvent yields marginally better results than saturating the first solvation shell around Co throughout. PBE performs best, displaying balanced accuracy and superior performance overall, with RMS errors of 80 mV for seven reduction potentials, 2.0 log units for five pKas and 2.3 log units for two log Kon/off values for the aquacobalamin system. Furthermore, we find that the BP86 functional commonly used in corrinoid studies suffers from erratic behavior and inaccurate descriptions of Co-axial ligand binding, leading to substantial errors in predicted pKas and

  13. Toward Quantitatively Accurate Calculation of the Redox-Associated Acid-Base and Ligand Binding Equilibria of Aquacobalamin.

    PubMed

    Johnston, Ryne C; Zhou, Jing; Smith, Jeremy C; Parks, Jerry M

    2016-08-01

    Redox processes in complex transition metal-containing species are often intimately associated with changes in ligand protonation states and metal coordination number. A major challenge is therefore to develop consistent computational approaches for computing pH-dependent redox and ligand dissociation properties of organometallic species. Reduction of the Co center in the vitamin B12 derivative aquacobalamin can be accompanied by ligand dissociation, protonation, or both, making these properties difficult to compute accurately. We examine this challenge here by using density functional theory and continuum solvation to compute Co-ligand binding equilibrium constants (Kon/off), pKas, and reduction potentials for models of aquacobalamin in aqueous solution. We consider two models for cobalamin ligand coordination: the first follows the hexa, penta, tetra coordination scheme for Co(III), Co(II), and Co(I) species, respectively, and the second model features saturation of each vacant axial coordination site on Co(II) and Co(I) species with a single, explicit water molecule to maintain six directly interacting ligands or water molecules in each oxidation state. Comparing these two coordination schemes in combination with five dispersion-corrected density functionals, we find that the accuracy of the computed properties is largely independent of the scheme used, but including only a continuum representation of the solvent yields marginally better results than saturating the first solvation shell around Co throughout. PBE performs best, displaying balanced accuracy and superior performance overall, with RMS errors of 80 mV for seven reduction potentials, 2.0 log units for five pKas and 2.3 log units for two log Kon/off values for the aquacobalamin system. Furthermore, we find that the BP86 functional commonly used in corrinoid studies suffers from erratic behavior and inaccurate descriptions of Co-axial ligand binding, leading to substantial errors in predicted pKas and

  14. Fast and accurate procedure for the determination of Cr(VI) in solid samples by isotope dilution mass spectrometry.

    PubMed

    Fabregat-Cabello, Neus; Rodríguez-González, Pablo; Castillo, Ángel; Malherbe, Julien; Roig-Navarro, Antoni F; Long, Stephen E; García Alonso, J Ignacio

    2012-11-20

    We present here a new environmental measurement method for the rapid extraction and accurate quantification of Cr(VI) in solid samples. The quantitative extraction of Cr(VI) is achieved in 10 minutes by means of focused microwave assisted extraction using 50 mmol/L Ethylendiamintetraacetic acid (EDTA) at pH 10 as extractant. In addition, it enables the separation of Cr species by anion exchange chromatography using a mobile phase which is a 1:10 dilution of the extracting solution. Thus, neutralization or acidification steps which are prone to cause interconversion of Cr species are not needed. Another benefit of using EDTA is that it allows to measure Cr(III)-EDTA complex and Cr(VI) simultaneously in an alkaline extraction solution. The application of a 10 minutes focused microwave assisted extraction (5 min at 90 °C plus 5 min at 110 °C) has been shown to quantitatively extract all forms of hexavalent chromium from the standard reference materials (SRM) candidate NIST 2700 and NIST 2701. A double spike isotope dilution mass spectrometry (IDMS) procedure was employed to study chromium interconversion reactions. It was observed that the formation of a Cr(III)-EDTA complex avoided Cr(III) oxidation for these two reference materials. Thus, the use of a double spiking strategy for quantification is not required and a single spike IDMS procedure using isotopically enriched Cr(VI) provided accurate results.

  15. Effect of sampling size on the determination of accurate pesticide residue levels in Japanese agricultural commodities.

    PubMed

    Fujita, Masahiro; Yajima, Tomonari; Iijima, Kazuaki; Sato, Kiyoshi

    2012-05-01

    The uncertainty in pesticide residue levels (UPRL) associated with sampling size was estimated using individual acetamiprid and cypermethrin residue data from preharvested apple, broccoli, cabbage, grape, and sweet pepper samples. The relative standard deviation from the mean of each sampling size (n = 2(x), where x = 1-6) of randomly selected samples was defined as the UPRL for each sampling size. The estimated UPRLs, which were calculated on the basis of the regulatory sampling size recommended by the OECD Guidelines on Crop Field Trials (weights from 1 to 5 kg, and commodity unit numbers from 12 to 24), ranged from 2.1% for cypermethrin in sweet peppers to 14.6% for cypermethrin in cabbage samples. The percentages of commodity exceeding the maximum residue limits (MRLs) specified by the Japanese Food Sanitation Law may be predicted from the equation derived from this study, which was based on samples of various size ranges with mean residue levels below the MRL. The estimated UPRLs have confirmed that sufficient sampling weight and numbers are required for analysis and/or re-examination of subsamples to provide accurate values of pesticide residue levels for the enforcement of MRLs. The equation derived from the present study would aid the estimation of more accurate residue levels even from small sampling sizes. PMID:22475588

  16. Accurate label-free reaction kinetics determination using initial rate heat measurements.

    PubMed

    Ebrahimi, Kourosh Honarmand; Hagedoorn, Peter-Leon; Jacobs, Denise; Hagen, Wilfred R

    2015-01-01

    Accurate label-free methods or assays to obtain the initial reaction rates have significant importance in fundamental studies of enzymes and in application-oriented high throughput screening of enzyme activity. Here we introduce a label-free approach for obtaining initial rates of enzyme activity from heat measurements, which we name initial rate calorimetry (IrCal). This approach is based on our new finding that the data recorded by isothermal titration calorimetry for the early stages of a reaction, which have been widely ignored, are correlated to the initial rates. Application of the IrCal approach to various enzymes led to accurate enzyme kinetics parameters as compared to spectroscopic methods and enabled enzyme kinetic studies with natural substrate, e.g. proteases with protein substrates. Because heat is a label-free property of almost all reactions, the IrCal approach holds promise in fundamental studies of various enzymes and in use of calorimetry for high throughput screening of enzyme activity.

  17. Accurate label-free reaction kinetics determination using initial rate heat measurements

    PubMed Central

    Ebrahimi, Kourosh Honarmand; Hagedoorn, Peter-Leon; Jacobs, Denise; Hagen, Wilfred R.

    2015-01-01

    Accurate label-free methods or assays to obtain the initial reaction rates have significant importance in fundamental studies of enzymes and in application-oriented high throughput screening of enzyme activity. Here we introduce a label-free approach for obtaining initial rates of enzyme activity from heat measurements, which we name initial rate calorimetry (IrCal). This approach is based on our new finding that the data recorded by isothermal titration calorimetry for the early stages of a reaction, which have been widely ignored, are correlated to the initial rates. Application of the IrCal approach to various enzymes led to accurate enzyme kinetics parameters as compared to spectroscopic methods and enabled enzyme kinetic studies with natural substrate, e.g. proteases with protein substrates. Because heat is a label-free property of almost all reactions, the IrCal approach holds promise in fundamental studies of various enzymes and in use of calorimetry for high throughput screening of enzyme activity. PMID:26574737

  18. Determination of infectious retrovirus concentration from colony-forming assay with quantitative analysis.

    PubMed

    Kwon, Young Jik; Hung, Gene; Anderson, W French; Peng, Ching-An; Yu, Hong

    2003-05-01

    The colony formation assay is the most commonly used titration method for defining the concentration of replication-incompetent murine leukemia virus-derived retroviral vectors. However, titer varies with target cell type and number, transduction time, and concentration of polycation (e.g., Polybrene). Moreover, because most of the viruses cannot encounter target cells due to Brownian motion, their short half-lives, and the requirement for target cell division for activity, the actual infectious retrovirus concentration in the collected supernatant is higher than the viral titer. Here we correlate the physical viral particle concentration with the infectious virus concentration and colony formation titer with the help of a mathematical model. Ecotropic murine leukemia retrovirus supernatant, collected from the GP+E86/LNCX retroviral vector producer cell line, was concentrated by centrifugation and further purified by a sucrose density gradient. The physical concentration of purified viral vectors was determined by direct particle counting with an electron microscope. The concentrations of fresh and concentrated supernatant were determined by a quantitative reverse transcriptase activity assay. Titration of all supernatants by neomycin-resistant colony formation assay was also performed. There were 767 +/- 517 physical viral particles per infectious CFU in the crude viral supernatant. However, the infectious viral concentration determined by mathematical simulation was 143 viral particles per infectious unit, which is more consistent with the concentration determined by particle counting in purified viral solution. Our results suggest that the mathematical model can be used to extract a more accurate and reliable concentration of infectious retrovirus.

  19. Quantitative determination of phenol by high-pressure liquid chromatography.

    PubMed

    Musto, J D; Sane, J N; Warner, V D

    1977-08-01

    High-pressure liquid chromatography was used with a 5-micrometer silica gel column to quantitate the phenol in phenolated calamine lotion USP and a commercial antiseptic solution. This method requires less than 10 min/assay, and other compounds present in the products analyzed did not interfere.

  20. Quantitative determination of phenol in phenolated calamine lotion USP.

    PubMed

    Das Gupta, V; Bomer, K A

    1975-07-01

    A method for the quantitative analysis of phenol in phenolated calamine lotion USP is described. The method is based on spectrophotometrically measuring the color produced by reacting phenol with either ferric chloride or ferric nitrate. Beer's law is followed. The effect of ferric-ion concentration on the sensitivity of the assay method is reported.

  1. Quantitive determination of flavin nucleotide using the bacterial bioluminescent reaction

    NASA Technical Reports Server (NTRS)

    Chappelle, E. W.; Picciolo, G. L.

    1969-01-01

    Photometric method based on the use of bacterial luminiscent reaction quantitatively detects the presence of flavin compounds in all forms of life. Aqueous cellular dispersion of a biological sample with an aqueous perchloric acid ruptures the cells and frees the flavin coenzymes from their proteins.

  2. Rigour in quantitative research.

    PubMed

    Claydon, Leica Sarah

    2015-07-22

    This article which forms part of the research series addresses scientific rigour in quantitative research. It explores the basis and use of quantitative research and the nature of scientific rigour. It examines how the reader may determine whether quantitative research results are accurate, the questions that should be asked to determine accuracy and the checklists that may be used in this process. Quantitative research has advantages in nursing, since it can provide numerical data to help answer questions encountered in everyday practice.

  3. Identification and validation of reference genes for accurate normalization of real-time quantitative PCR data in kiwifruit.

    PubMed

    Ferradás, Yolanda; Rey, Laura; Martínez, Óscar; Rey, Manuel; González, Ma Victoria

    2016-05-01

    Identification and validation of reference genes are required for the normalization of qPCR data. We studied the expression stability produced by eight primer pairs amplifying four common genes used as references for normalization. Samples representing different tissues, organs and developmental stages in kiwifruit (Actinidia chinensis var. deliciosa (A. Chev.) A. Chev.) were used. A total of 117 kiwifruit samples were divided into five sample sets (mature leaves, axillary buds, stigmatic arms, fruit flesh and seeds). All samples were also analysed as a single set. The expression stability of the candidate primer pairs was tested using three algorithms (geNorm, NormFinder and BestKeeper). The minimum number of reference genes necessary for normalization was also determined. A unique primer pair was selected for amplifying the 18S rRNA gene. The primer pair selected for amplifying the ACTIN gene was different depending on the sample set. 18S 2 and ACT 2 were the candidate primer pairs selected for normalization in the three sample sets (mature leaves, fruit flesh and stigmatic arms). 18S 2 and ACT 3 were the primer pairs selected for normalization in axillary buds. No primer pair could be selected for use as the reference for the seed sample set. The analysis of all samples in a single set did not produce the selection of any stably expressing primer pair. Considering data previously reported in the literature, we validated the selected primer pairs amplifying the FLOWERING LOCUS T gene for use in the normalization of gene expression in kiwifruit.

  4. Identification and validation of reference genes for accurate normalization of real-time quantitative PCR data in kiwifruit.

    PubMed

    Ferradás, Yolanda; Rey, Laura; Martínez, Óscar; Rey, Manuel; González, Ma Victoria

    2016-05-01

    Identification and validation of reference genes are required for the normalization of qPCR data. We studied the expression stability produced by eight primer pairs amplifying four common genes used as references for normalization. Samples representing different tissues, organs and developmental stages in kiwifruit (Actinidia chinensis var. deliciosa (A. Chev.) A. Chev.) were used. A total of 117 kiwifruit samples were divided into five sample sets (mature leaves, axillary buds, stigmatic arms, fruit flesh and seeds). All samples were also analysed as a single set. The expression stability of the candidate primer pairs was tested using three algorithms (geNorm, NormFinder and BestKeeper). The minimum number of reference genes necessary for normalization was also determined. A unique primer pair was selected for amplifying the 18S rRNA gene. The primer pair selected for amplifying the ACTIN gene was different depending on the sample set. 18S 2 and ACT 2 were the candidate primer pairs selected for normalization in the three sample sets (mature leaves, fruit flesh and stigmatic arms). 18S 2 and ACT 3 were the primer pairs selected for normalization in axillary buds. No primer pair could be selected for use as the reference for the seed sample set. The analysis of all samples in a single set did not produce the selection of any stably expressing primer pair. Considering data previously reported in the literature, we validated the selected primer pairs amplifying the FLOWERING LOCUS T gene for use in the normalization of gene expression in kiwifruit. PMID:26897117

  5. [A simple test for quantitative determination of LDL-cholesterol].

    PubMed

    Mertz, D P; Thuilot, G

    1986-05-01

    The subject of the report is a novel precipitation test for the quantitative recording of LDL cholesterol based on the precipitation of LDL by dextran sulphate. Parallel assays of LDL cholesterol according to the new method and using quantitative lipoprotein electrophoresis as reference showed the results, in terms of the individual values and collectively, to be practically identical for a wide concentration range of various lipids and lipoproteins in the serum. The concentration ratio of the means obtained according to the two methods is 1.014 +/- 0.102 (standard deviation). The regression function displays a correlation coefficient of 0.9470. Double assays with the new technique yield a variation coefficient of 1.7 +/- 0.4%. Limitations of the method, which are insignificant for application in practice, are pointed out. The new precipitation method is simple, safe and useful for the quantitative estimation of the LDL cholesterol concentration in freshly obtained human serum. The method requires only little time and equipment.

  6. Simultaneous quantitative determination of paracetamol and tramadol in tablet formulation using UV spectrophotometry and chemometric methods.

    PubMed

    Glavanović, Siniša; Glavanović, Marija; Tomišić, Vladislav

    2016-03-15

    The UV spectrophotometric methods for simultaneous quantitative determination of paracetamol and tramadol in paracetamol-tramadol tablets were developed. The spectrophotometric data obtained were processed by means of partial least squares (PLS) and genetic algorithm coupled with PLS (GA-PLS) methods in order to determine the content of active substances in the tablets. The results gained by chemometric processing of the spectroscopic data were statistically compared with those obtained by means of validated ultra-high performance liquid chromatographic (UHPLC) method. The accuracy and precision of data obtained by the developed chemometric models were verified by analysing the synthetic mixture of drugs, and by calculating recovery as well as relative standard error (RSE). A statistically good agreement was found between the amounts of paracetamol determined using PLS and GA-PLS algorithms, and that obtained by UHPLC analysis, whereas for tramadol GA-PLS results were proven to be more reliable compared to those of PLS. The simplest and the most accurate and precise models were constructed by using the PLS method for paracetamol (mean recovery 99.5%, RSE 0.89%) and the GA-PLS method for tramadol (mean recovery 99.4%, RSE 1.69%).

  7. Simultaneous quantitative determination of paracetamol and tramadol in tablet formulation using UV spectrophotometry and chemometric methods

    NASA Astrophysics Data System (ADS)

    Glavanović, Siniša; Glavanović, Marija; Tomišić, Vladislav

    2016-03-01

    The UV spectrophotometric methods for simultaneous quantitative determination of paracetamol and tramadol in paracetamol-tramadol tablets were developed. The spectrophotometric data obtained were processed by means of partial least squares (PLS) and genetic algorithm coupled with PLS (GA-PLS) methods in order to determine the content of active substances in the tablets. The results gained by chemometric processing of the spectroscopic data were statistically compared with those obtained by means of validated ultra-high performance liquid chromatographic (UHPLC) method. The accuracy and precision of data obtained by the developed chemometric models were verified by analysing the synthetic mixture of drugs, and by calculating recovery as well as relative standard error (RSE). A statistically good agreement was found between the amounts of paracetamol determined using PLS and GA-PLS algorithms, and that obtained by UHPLC analysis, whereas for tramadol GA-PLS results were proven to be more reliable compared to those of PLS. The simplest and the most accurate and precise models were constructed by using the PLS method for paracetamol (mean recovery 99.5%, RSE 0.89%) and the GA-PLS method for tramadol (mean recovery 99.4%, RSE 1.69%).

  8. Accurate determination of the binding energy of the formic acid dimer: the importance of geometry relaxation.

    PubMed

    Kalescky, Robert; Kraka, Elfi; Cremer, Dieter

    2014-02-28

    The formic acid dimer in its C2h-symmetrical cyclic form is stabilized by two equivalent H-bonds. The currently accepted interaction energy is 18.75 kcal/mol whereas the experimental binding energy D0 value is only 14.22 ±0.12 kcal/mol [F. Kollipost, R. W. Larsen, A. V. Domanskaya, M. Nörenberg, and M. A. Suhm, J. Chem. Phys. 136, 151101 (2012)]. Calculation of the binding energies De and D0 at the CCSD(T) (Coupled Cluster with Single and Double excitations and perturbative Triple excitations)/CBS (Complete Basis Set) level of theory, utilizing CCSD(T)/CBS geometries and the frequencies of the dimer and monomer, reveals that there is a 3.2 kcal/mol difference between interaction energy and binding energy De, which results from (i) not relaxing the geometry of the monomers upon dissociation of the dimer and (ii) approximating CCSD(T) correlation effects with MP2. The most accurate CCSD(T)/CBS values obtained in this work are De = 15.55 and D0 = 14.32 kcal/mol where the latter binding energy differs from the experimental value by 0.1 kcal/mol. The necessity of employing augmented VQZ and VPZ calculations and relaxing monomer geometries of H-bonded complexes upon dissociation to obtain reliable binding energies is emphasized.

  9. Accurate determination of the binding energy of the formic acid dimer: The importance of geometry relaxation

    NASA Astrophysics Data System (ADS)

    Kalescky, Robert; Kraka, Elfi; Cremer, Dieter

    2014-02-01

    The formic acid dimer in its C2h-symmetrical cyclic form is stabilized by two equivalent H-bonds. The currently accepted interaction energy is 18.75 kcal/mol whereas the experimental binding energy D0 value is only 14.22 ±0.12 kcal/mol [F. Kollipost, R. W. Larsen, A. V. Domanskaya, M. Nörenberg, and M. A. Suhm, J. Chem. Phys. 136, 151101 (2012)]. Calculation of the binding energies De and D0 at the CCSD(T) (Coupled Cluster with Single and Double excitations and perturbative Triple excitations)/CBS (Complete Basis Set) level of theory, utilizing CCSD(T)/CBS geometries and the frequencies of the dimer and monomer, reveals that there is a 3.2 kcal/mol difference between interaction energy and binding energy De, which results from (i) not relaxing the geometry of the monomers upon dissociation of the dimer and (ii) approximating CCSD(T) correlation effects with MP2. The most accurate CCSD(T)/CBS values obtained in this work are De = 15.55 and D0 = 14.32 kcal/mol where the latter binding energy differs from the experimental value by 0.1 kcal/mol. The necessity of employing augmented VQZ and VPZ calculations and relaxing monomer geometries of H-bonded complexes upon dissociation to obtain reliable binding energies is emphasized.

  10. A colorimetric-based accurate method for the determination of enterovirus 71 titer.

    PubMed

    Pourianfar, Hamid Reza; Javadi, Arman; Grollo, Lara

    2012-12-01

    The 50 % tissue culture infectious dose (TCID50) is still one of the most commonly used techniques for estimating virus titers. However, the traditional TCID50 assay is time consuming, susceptible to subjective errors and generates only quantal data. Here, we describe a colorimetric-based approach for the titration of Enterovirus 71 (EV71) using a modified method for making virus dilutions. In summary, the titration of EV71 using MTT or MTS staining with a modified virus dilution method decreased the time of the assay and eliminated the subjectivity of observational results, improving accuracy, reproducibility and reliability of virus titration, in comparison with the conventional TCID50 approach (p < 0.01). In addition, the results provided evidence that there was better correlation between a plaquing assay and our approach when compared to the traditional TCID50 approach. This increased accuracy also improved the ability to predict the number of virus plaque forming units present in a solution. These improvements could be of use for any virological experimentation, where a quick accurate titration of a virus capable of causing cell destruction is required or a sensible estimation of the number of viral plaques based on TCID50 of a virus is desired.

  11. Do Ecological Niche Models Accurately Identify Climatic Determinants of Species Ranges?

    PubMed

    Searcy, Christopher A; Shaffer, H Bradley

    2016-04-01

    Defining species' niches is central to understanding their distributions and is thus fundamental to basic ecology and climate change projections. Ecological niche models (ENMs) are a key component of making accurate projections and include descriptions of the niche in terms of both response curves and rankings of variable importance. In this study, we evaluate Maxent's ranking of environmental variables based on their importance in delimiting species' range boundaries by asking whether these same variables also govern annual recruitment based on long-term demographic studies. We found that Maxent-based assessments of variable importance in setting range boundaries in the California tiger salamander (Ambystoma californiense; CTS) correlate very well with how important those variables are in governing ongoing recruitment of CTS at the population level. This strong correlation suggests that Maxent's ranking of variable importance captures biologically realistic assessments of factors governing population persistence. However, this result holds only when Maxent models are built using best-practice procedures and variables are ranked based on permutation importance. Our study highlights the need for building high-quality niche models and provides encouraging evidence that when such models are built, they can reflect important aspects of a species' ecology. PMID:27028071

  12. Spreading of liquid droplets on cylindrical surfaces: Accurate determination of contact angle

    NASA Astrophysics Data System (ADS)

    Wagner, H. D.

    1990-02-01

    The characterization of the physicochemical nature of interfaces is a key problem in the field of advanced fibrous composites. The macroscopic regime contact angle, which reflects the energetics of wetting at the solid-liquid interface, is difficult to measure by usual methods in the case of very thin cylindrical fibers, but it may be calculated from the shape of a liquid droplet spread onto a cylindrical monofilament using a method developed by Yamaki and Katayama [J. Appl. Polym. Sci. 19, 2897 (1975)], and B. J. Carroll [J. Coll. Interf. Sci. 57, 488 (1976)]. Unfortunately, measurements of the contact angle based on this method are, so far, unable to provide an accuracy of better than about 5°. In the present article two simple extensions of the method of Yamaki and Katayama and Carroll, are presented, from which highly accurate values of the contact angle may be obtained. This is demonstrated experimentally from the spreading of glycerol droplets on carbon fibers and epoxy droplets on aramid fibers.

  13. Variant of more accurate determination of the locations of shortwave radio emission sources

    NASA Astrophysics Data System (ADS)

    Ivanov, V. F.; Myslivtsev, T. O.; Troitskii, B. V.

    2013-04-01

    The paper discusses how the trajectory calculation method can be used to solve the problem of locality determination of shortwave (SW) emission sources. The dependence of the electron concentration on the coordinates is specified using the SPIM model; it is corrected using the ionospheric solar activity index, which is specified with the help of maps of total electron content. We suggested a variant of how a regional map of the total electron content can be plotted according to measurements of signals from GLONASS/GPS navigation systems. It is shown that the trajectory calculation method, coupled with an adjustable ionospheric model, allows for a more exact locality determination of SW radio emission sources.

  14. [Quantitative determination of penicillins by iodometry using potassium hydrogen peroxymonosulfate].

    PubMed

    Blazhevskiĭ, N E; Karpova, S P; Kabachyĭ, V I

    2013-01-01

    The kinetics and stoichiometry of S-oxidation of semisynthetic penicillins (amoxicillin trihydrate, ampicillin trihydrate, sodium salt of oxacillin and ticarcillin disodium salt) by potassium hydrogen peroxymonosulfate in aqueous solutions at pH 3-6 was studied by iodometric titration: 1 mol of KNSO5 per 1 mol of penicillin, the quantitative interaction is achieved in 1 min (time of observation). A unified method was developed and the possibility of quantification of penicillins by the iodometric method using potassium hydrogen peroxymonosulfate as an analytical reagent was shown.

  15. Molecular Simulation of the Free Energy for the Accurate Determination of Phase Transition Properties of Molecular Solids

    NASA Astrophysics Data System (ADS)

    Sellers, Michael; Lisal, Martin; Brennan, John

    2015-06-01

    Investigating the ability of a molecular model to accurately represent a real material is crucial to model development and use. When the model simulates materials in extreme conditions, one such property worth evaluating is the phase transition point. However, phase transitions are often overlooked or approximated because of difficulty or inaccuracy when simulating them. Techniques such as super-heating or super-squeezing a material to induce a phase change suffer from inherent timescale limitations leading to ``over-driving,'' and dual-phase simulations require many long-time runs to seek out what frequently results in an inexact location of phase-coexistence. We present a compilation of methods for the determination of solid-solid and solid-liquid phase transition points through the accurate calculation of the chemical potential. The methods are applied to the Smith-Bharadwaj atomistic potential's representation of cyclotrimethylene trinitramine (RDX) to accurately determine its melting point (Tm) and the alpha to gamma solid phase transition pressure. We also determine Tm for a coarse-grain model of RDX, and compare its value to experiment and atomistic counterpart. All methods are employed via the LAMMPS simulator, resulting in 60-70 simulations that total 30-50 ns. Approved for public release. Distribution is unlimited.

  16. Automated methods for accurate determination of the critical velocity of packed bed chromatography.

    PubMed

    Chang, Yu-Chih; Gerontas, Spyridon; Titchener-Hooker, Nigel J

    2012-01-01

    Knowing the critical velocity (ucrit) of a chromatography column is an important part of process development as it allows the optimization of chromatographic flow conditions. The conventional flow step method for determining ucrit is prone to error as it depends heavily on human judgment. In this study, two automated methods for determining ucrit have been developed: the automatic flow step (AFS) method and the automatic pressure step (APS) method. In the AFS method, the column pressure drop is monitored upon application of automated incremental increases in flow velocity, whereas in the APS method the flow velocity is monitored upon application of automated incremental increases in pressure drop. The APS method emerged as the one with the higher levels of accuracy, efficiency and ease of application having the greater potential to assist defining the best operational parameters of a chromatography column.

  17. Accurate determination of imaging modality using an ensemble of text- and image-based classifiers.

    PubMed

    Kahn, Charles E; Kalpathy-Cramer, Jayashree; Lam, Cesar A; Eldredge, Christina E

    2012-02-01

    Imaging modality can aid retrieval of medical images for clinical practice, research, and education. We evaluated whether an ensemble classifier could outperform its constituent individual classifiers in determining the modality of figures from radiology journals. Seventeen automated classifiers analyzed 77,495 images from two radiology journals. Each classifier assigned one of eight imaging modalities--computed tomography, graphic, magnetic resonance imaging, nuclear medicine, positron emission tomography, photograph, ultrasound, or radiograph-to each image based on visual and/or textual information. Three physicians determined the modality of 5,000 randomly selected images as a reference standard. A "Simple Vote" ensemble classifier assigned each image to the modality that received the greatest number of individual classifiers' votes. A "Weighted Vote" classifier weighted each individual classifier's vote based on performance over a training set. For each image, this classifier's output was the imaging modality that received the greatest weighted vote score. We measured precision, recall, and F score (the harmonic mean of precision and recall) for each classifier. Individual classifiers' F scores ranged from 0.184 to 0.892. The simple vote and weighted vote classifiers correctly assigned 4,565 images (F score, 0.913; 95% confidence interval, 0.905-0.921) and 4,672 images (F score, 0.934; 95% confidence interval, 0.927-0.941), respectively. The weighted vote classifier performed significantly better than all individual classifiers. An ensemble classifier correctly determined the imaging modality of 93% of figures in our sample. The imaging modality of figures published in radiology journals can be determined with high accuracy, which will improve systems for image retrieval.

  18. A rural hospital's use of software to determine accurate Medicare reimbursement levels.

    PubMed

    Weiszbrod, T

    1995-01-01

    This article details how a software program has helped a hospital collect more Medicare payments for which it was eligible than it previously had collected. The software simplifies the process of selecting complications by reminding the user when no complications have yet been entered, by determining whether each item entered qualifies as a complication, and by suggesting other possibilities. In many instances, these complications are possibilities that a person performing manual coding might not consider. PMID:10144983

  19. An accurate method for the determination of carboxyhemoglobin in postmortem blood using GC-TCD.

    PubMed

    Lewis, Russell J; Johnson, Robert D; Canfield, Dennis V

    2004-01-01

    During the investigation of aviation accidents, postmortem samples from accident victims are submitted to the FAA's Civil Aerospace Medical Institute for toxicological analysis. In order to determine if an accident victim was exposed to an in-flight/postcrash fire or faulty heating/exhaust system, the analysis of carbon monoxide (CO) is conducted. Although our laboratory predominantly uses a spectrophotometric method for the determination of carboxyhemoglobin (COHb), we consider it essential to confirm with a second technique based on a different analytical principle. Our laboratory encountered difficulties with many of our postmortem samples while employing a commonly used GC method. We believed these problems were due to elevated methemoglobin (MetHb) concentration in our specimens. MetHb does not bind CO; therefore, elevated MetHb levels will result in a loss of CO-binding capacity. Because most commonly employed GC methods determine %COHb from a ratio of unsaturated blood to CO-saturated blood, a loss of CO-binding capacity will result in an erroneously high %COHb value. Our laboratory has developed a new GC method for the determination of %COHb that incorporates sodium dithionite, which will reduce any MetHb present to Hb. Using blood controls ranging from 1% to 67% COHb, we found no statistically significant differences between %COHb results from our new GC method and our spectrophotometric method. To validate the new GC method, postmortem samples were analyzed with our existing spectrophotometric method, a GC method commonly used without reducing agent, and our new GC method with the addition of sodium dithionite. As expected, we saw errors up to and exceeding 50% when comparing the unreduced GC results with our spectrophotometric method. With our new GC procedure, the error was virtually eliminated. PMID:14987426

  20. Determination of the biotin content of select foods using accurate and sensitive HPLC/avidin binding

    PubMed Central

    Staggs, C.G.; Sealey, W.M.; McCabe, B.J.; Teague, A.M.; Mock, D.M.

    2006-01-01

    Assessing dietary biotin content, biotin bioavailability, and resulting biotin status are crucial in determining whether biotin deficiency is teratogenic in humans. Accuracy in estimating dietary biotin is limited both by data gaps in food composition tables and by inaccuracies in published data. The present study applied sensitive and specific analytical techniques to determine values for biotin content in a select group of foods. Total biotin content of 87 foods was determined using acid hydrolysis and the HPLC/avidin-binding assay. These values are consistent with published values in that meat, fish, poultry, egg, dairy, and some vegetables are relatively rich sources of biotin. However, these biotin values disagreed substantially with published values for many foods. Assay values varied between 247 times greater than published values for a given food to as much as 36% less than the published biotin value. Among 51 foods assayed for which published values were available, only seven agreed within analytical variability (720%). We conclude that published values for biotin content of foods are likely to be inaccurate. PMID:16648879

  1. Use of quantitative shape-activity relationships to model the photoinduced toxicity of polycyclic aromatic hydrocarbons: Electron density shape features accurately predict toxicity

    SciTech Connect

    Mezey, P.G.; Zimpel, Z.; Warburton, P.; Walker, P.D.; Irvine, D.G.; Huang, X.D.; Dixon, D.G.; Greenberg, B.M.

    1998-07-01

    The quantitative shape-activity relationship (QShAR) methodology, based on accurate three-dimensional electron densities and detailed shape analysis methods, has been applied to a Lemna gibba photoinduced toxicity data set of 16 polycyclic aromatic hydrocarbon (PAH) molecules. In the first phase of the studies, a shape fragment QShAR database of PAHs was developed. The results provide a very good match to toxicity based on a combination of the local shape features of single rings in comparison to the central ring of anthracene and a more global shape feature involving larger molecular fragments. The local shape feature appears as a descriptor of the susceptibility of PAHs to photomodification and the global shape feature is probably related to photosensitization activity.

  2. Procedure for the quantitative determination of mixtures of nucleic Acid components based on multivariate spectrophotometric Acid-base titrations.

    PubMed

    Saurina, J; Hernández-Cassou, S; Tauler, R; Izquierdo-Ridorsa, A

    1999-01-01

    A new procedure for the quantitative determination of mixtures of nucleic acid components, based on continuous spectrophotometric acid-base titrations and multivariate curve resolution, is proposed. The procedure simultaneously takes into account the spectroscopic and acid-base properties of the compounds, which leads to a higher selectivity. Furthermore, quantitative determination of an analyte in a complex mixture is performed using a synthetic solution as standard containing only the analyte of interest. An intrinsic difficulty in the analysis of spectrometric titration data is the presence of rank deficiency due to closure for the mixtures of two or more compounds. An additional problem can be encountered in some mixtures if species spectra or species concentration profiles are practically identical (rank overlap). However, even in the presence of these rank difficulties, accurate quantitation with prediction errors lower than 5% was obtained. The presence of unknown and uncalibrated interferences in the samples does not affect the quantitative determination of the analyte of interest. The proposed procedure was successfully applied to the analysis of real samples (pharmaceuticals) using synthetic external standards.

  3. Statistically deformable 2D/3D registration for accurate determination of post-operative cup orientation from single standard X-ray radiograph.

    PubMed

    Zheng, Guoyan

    2009-01-01

    The widely used procedure of evaluation of cup orientation following total hip arthroplasty using single standard anteroposterior (AP) radiograph is known inaccurate, largely due to the wide variability in individual pelvic orientation relative to X-ray plate. 2D/3D rigid image registration methods have been introduced for an accurate determination of the post-operative cup alignment with respect to an anatomical reference extracted from the CT data. Although encouraging results have been reported, their extensive usage in clinical routine is still limited. This may be explained by their requirement of a CAD model of the prosthesis, which is often difficult to be organized from the manufacturer due to the proprietary issue, and by their requirement of a pre-operative CT scan, which is not available for most retrospective studies. To address these issues, we developed and validated a statistically deformable 2D/3D registration approach for accurate determination of post-operative cup orientation. No CAD model and pre-operative CT data is required any more. Quantitative and qualitative results evaluated on cadaveric and clinical datasets are given, which indicate the validity of the approach. PMID:20426064

  4. Raman Spectroscopy Provides a Powerful Diagnostic Tool for Accurate Determination of Albumin Glycation

    PubMed Central

    Dingari, Narahara Chari; Horowitz, Gary L.; Kang, Jeon Woong; Dasari, Ramachandra R.; Barman, Ishan

    2012-01-01

    We present the first demonstration of glycated albumin detection and quantification using Raman spectroscopy without the addition of reagents. Glycated albumin is an important marker for monitoring the long-term glycemic history of diabetics, especially as its concentrations, in contrast to glycated hemoglobin levels, are unaffected by changes in erythrocyte life times. Clinically, glycated albumin concentrations show a strong correlation with the development of serious diabetes complications including nephropathy and retinopathy. In this article, we propose and evaluate the efficacy of Raman spectroscopy for determination of this important analyte. By utilizing the pre-concentration obtained through drop-coating deposition, we show that glycation of albumin leads to subtle, but consistent, changes in vibrational features, which with the help of multivariate classification techniques can be used to discriminate glycated albumin from the unglycated variant with 100% accuracy. Moreover, we demonstrate that the calibration model developed on the glycated albumin spectral dataset shows high predictive power, even at substantially lower concentrations than those typically encountered in clinical practice. In fact, the limit of detection for glycated albumin measurements is calculated to be approximately four times lower than its minimum physiological concentration. Importantly, in relation to the existing detection methods for glycated albumin, the proposed method is also completely reagent-free, requires barely any sample preparation and has the potential for simultaneous determination of glycated hemoglobin levels as well. Given these key advantages, we believe that the proposed approach can provide a uniquely powerful tool for quantification of glycation status of proteins in biopharmaceutical development as well as for glycemic marker determination in routine clinical diagnostics in the future. PMID:22393405

  5. Accurate mass determination of short-lived isotopes by a tandem Penning-trap mass spectrometer

    SciTech Connect

    Stolzenberg, H.; Becker, S.; Bollen, G.; Kern, F.; Kluge, H.; Otto, T.; Savard, G.; Schweikhard, L. ); Audi, G. ); Moore, R.B. ); The ISOLDE Collaboration

    1990-12-17

    A mass spectrometer consisting of two Penning traps has been set up for short-lived isotopes at the on-line mass separator ISOLDE at CERN. The ion beam is collected and cooled in the first trap. After delivery to the second trap, high-accuracy direct mass measurements are made by determining the cyclotron frequency of the stored ions. Measurements have been performed for {sup 118}Cs--{sup 137}Cs. A resolving power of over 10{sup 6} and an accuracy of 1.4{times}10{sup {minus}7} have been achieved, corresponding to about 20 keV.

  6. Photo-detachment signal analysis to accurately determine electronegativity, electron temperature, and charged species density

    NASA Astrophysics Data System (ADS)

    Oudini, N.; Sirse, N.; Taccogna, F.; Ellingboe, A. R.; Bendib, A.

    2016-09-01

    Laser pulse induced photo-detachment combined with Langmuir probing has been introduced to diagnose plasma electronegativity. This technique uses a laser pulse to convert negative ions into electron-atom pairs and tracks the change of electron saturation current by a Langmuir probe. The existing model determines plasma electronegativity as the ratio of electron saturation current before and after detachment. However, this model depends on various assumptions and neglects the formation of a potential barrier between the laser channel and surrounding electronegative plasma. In this letter, we present a new analytical model to analyze photo-detachment signals in order to improve the accuracy of electronegativity measurements and extend this technique for measuring electron temperature and charged species density. This analytical model is supported by Particle-In-Cell simulation of electronegative plasma dynamics following laser photo-detachment. The analysis of the signal, detected on a simulated probe, shows that the present analytical model determines electronegativity, electron temperature, and plasma density with a relative error of ˜20%, ˜20%, and ˜50%, respectively, whereas the electronegativity obtained from a previous model is underestimated by an order of magnitude.

  7. Optical aperture area determination for accurate illuminance and luminous efficacy measurements of LED lamps

    NASA Astrophysics Data System (ADS)

    Dönsberg, Timo; Mäntynen, Henrik; Ikonen, Erkki

    2016-06-01

    The measurement uncertainty of illuminance and, consequently, luminous flux and luminous efficacy of LED lamps can be reduced with a recently introduced method based on the predictable quantum efficient detector (PQED). One of the most critical factors affecting the measurement uncertainty with the PQED method is the determination of the aperture area. This paper describes an upgrade to an optical method for direct determination of aperture area where superposition of equally spaced Gaussian laser beams is used to form a uniform irradiance distribution. In practice, this is accomplished by scanning the aperture in front of an intensity-stabilized laser beam. In the upgraded method, the aperture is attached to the PQED and the whole package is transversely scanned relative to the laser beam. This has the benefit of having identical geometry in the laser scanning of the aperture area and in the actual photometric measurement. Further, the aperture and detector assembly does not have to be dismantled for the aperture calibration. However, due to small acceptance angle of the PQED, differences between the diffraction effects of an overfilling plane wave and of a combination of Gaussian laser beams at the circular aperture need to be taken into account. A numerical calculation method for studying these effects is discussed in this paper. The calculation utilizes the Rayleigh-Sommerfeld diffraction integral, which is applied to the geometry of the PQED and the aperture. Calculation results for various aperture diameters and two different aperture-to-detector distances are presented.

  8. A new sensor system for accurate and precise determination of sediment dynamics and position.

    NASA Astrophysics Data System (ADS)

    Maniatis, Georgios; Hoey, Trevor; Sventek, Joseph; Hodge, Rebecca

    2014-05-01

    Sediment transport processes control many significant geomorphological changes. Consequently, sediment transport dynamics are studied across a wide range of scales leading to application of a variety of conceptually different mathematical descriptions (models) and data acquisition techniques (sensing). For river sediment transport processes both Eulerian and Lagrangian formulations are used. Data are gathered using a very wide range of sensing techniques that are not always compatible with the conceptual formulation applied. We are concerned with small to medium sediment grain-scale motion in gravel-bed rivers, and other coarse-grained environments, and: a) are developing a customised environmental sensor capable of providing coherent data that reliably record the motion; and, b) provide a mathematical framework in which these data can be analysed and interpreted, this being compatible with current stochastic approaches to sediment transport theory. Here we present results from three different aspects of the above developmental process. Firstly, we present a requirement analysis for the sensor based on the state of the art of the existing technologies. We focus on the factors that enhance data coherence and representativeness, extending the common practice for optimization which is based exclusively on electronics/computing related criteria. This analysis leads to formalization of a method that permits accurate control on the physical properties of the sensor using contemporary rapid prototyping techniques [Maniatis et al. 2013]. Secondly the first results are presented from a series of entrainment experiments in a 5 x 0.8 m flume in which a prototype sensor was deployed to monitor entrainment dynamics under increasing flow conditions (0.037 m3.s-1). The sensor was enclosed in an idealized spherical case (111 mm diameter) and placed on a constructed bed of hemispheres of the same diameter. We measured 3-axial inertial acceleration (as a measure of flow stress

  9. A Turn-On Resonance Raman Scattering (BCS/Cu+) Sensor for Quantitative Determination of Proteins.

    PubMed

    Chen, Lei; Xue, Xiangxin; Jiang, Dayu; Yang, Jin; Zhao, Bing; Han, Xiao Xia; Mee Jung, Young

    2016-02-01

    In this study, a new method for the quantitative evaluation of proteins is developed using competitive resonance Raman spectroscopy. A chelation reaction between bathocuproine disulfonate (BCS) and Cu(+) which is reduced by proteins in an alkaline environment, is utilized to create a BCS-Cu(+) complex that has strong resonance Raman activity. As a result, the method presented here enables protein detection over a much wider concentration range than conventional methods. Furthermore, the resonance Raman-based method can provide an improved lower limit of detection (500 pg/mL) compared to that obtained by colorimetric and ultraviolet- (UV-) based methods. Additionally, protein-to-protein variation can be determined using a standard curve created from known concentrations of bovine serum albumin (BSA), and excellent protein recovery is observed. More importantly, the proposed method was employing to the real sample (fetal bovine serum [FBS]). Based on the calibration curve from this method, it is extremely accurate for the determination of total protein concentrations in FBS. Results of this study demonstrate that the proposed method provides a novel and highly sensitive protocol for reagent-based protein assays.

  10. Accurate determination of the nuclear quadrupole moment of xenon from the molecular method

    NASA Astrophysics Data System (ADS)

    Canella, Guilherme A.; Santiago, Régis T.; Haiduke, Roberto L. A.

    2016-09-01

    This study provides a new determination of the nuclear electric quadrupole moment (NQM) for 131Xe, which is achieved by the molecular method. Dirac-Coulomb Coupled Cluster calculations with a Gaunt correction (DC+G-CC) of electric field gradients (EFGs) and experimental nuclear quadrupole coupling constants of six molecular systems (XeH+, XeCuF, XeCuCl, XeAgF, XeAgCl and XeAuF) were considered. The best NQM obtained by our DC+G-CCSD-T EFGs was -114.6(1.1) mbarn, which is recommended as the new reference value for this nuclide given the high level electron structure calculations done here.

  11. An experimental correction proposed for an accurate determination of mass diffusivity of wood in steady regime

    NASA Astrophysics Data System (ADS)

    Zohoun, Sylvain; Agoua, Eusèbe; Degan, Gérard; Perre, Patrick

    2002-08-01

    This paper presents an experimental study of the mass diffusion in the hygroscopic region of four temperate species and three tropical ones. In order to simplify the interpretation of the phenomena, a dimensionless parameter called reduced diffusivity is defined. This parameter varies from 0 to 1. The method used is firstly based on the determination of that parameter from results of the measurement of the mass flux which takes into account the conditions of operating standard device (tightness, dimensional variations and easy installation of samples of wood, good stability of temperature and humidity). Secondly the reasons why that parameter has to be corrected are presented. An abacus for this correction of mass diffusivity of wood in steady regime has been plotted. This work constitutes an advanced deal nowadays for characterising forest species.

  12. Accurate determination of silver nanoparticles in animal tissues by inductively coupled plasma mass spectrometry

    NASA Astrophysics Data System (ADS)

    Veverková, Lenka; Hradilová, Šárka; Milde, David; Panáček, Aleš; Skopalová, Jana; Kvítek, Libor; Petrželová, Kamila; Zbořil, Radek

    2014-12-01

    This study examined recoveries of silver determination in animal tissues after wet digestion by inductively coupled plasma mass spectrometry. The composition of the mineralization mixture for microwave assisted digestion was optimized and the best recoveries were obtained for mineralization with HNO3 and addition of HCl promptly after digestion. The optimization was performed on model samples of chicken meat spiked with silver nanoparticles and a solution of ionic silver. Basic calculations of theoretical distribution of Ag among various silver-containing species were implemented and the results showed that most of the silver is in the form of soluble complexes AgCl2- and AgCl32 - for the optimized composition of the mineralization mixture. Three animal tissue certified reference materials were then analyzed to verify the trueness and precision of the results.

  13. A new trend to determine biochemical parameters by quantitative FRET assays

    PubMed Central

    Liao, Jia-yu; Song, Yang; Liu, Yan

    2015-01-01

    Förster resonance energy transfer (FRET) has been widely used in biological and biomedical research because it can determine molecule or particle interactions within a range of 1–10 nm. The sensitivity and efficiency of FRET strongly depend on the distance between the FRET donor and acceptor. Historically, FRET assays have been used to quantitatively deduce molecular distances. However, another major potential application of the FRET assay has not been fully exploited, that is, the use of FRET signals to quantitatively describe molecular interactive events. In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (Kd), enzymatic velocity (kcat) and Km. We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution. PMID:26567729

  14. Quantitative Determination of DNA-Ligand Binding Using Fluorescence Spectroscopy

    ERIC Educational Resources Information Center

    Healy, Eamonn F.

    2007-01-01

    The effective use of fluorescence spectroscopy for determining the binding of the intercalcating agent crhidium bromide to DNA is being described. The analysis used simple measurement techniques and hence can be easily adopted by the students for a better understanding.

  15. Accurate Determination of Interstrand Distances and Alignment in Amyloid Fibrils by Magic Angle Spinning NMR

    PubMed Central

    Caporini, Marc A.; Bajaj, Vikram S.; Veshtort, Mikhail; Fitzpatrick, Anthony; MacPhee, Cait E; Vendruscolo, Michele; Dobson, Christopher M.; Griffin, Robert G.

    2010-01-01

    Amyloid fibrils are structurally ordered aggregates of proteins whose formation is associated with many neurodegenerative and other diseases. For that reason, their high resolution structures are of considerable interest and have been studied using a wide range of techniques, notably electron microscopy, x-ray diffraction, and magic angle spinning (MAS) NMR. Because of the excellent resolution in the spectra, MAS NMR is uniquely capable of delivering site-specific, atomic resolution information about all levels of amyloid structure: (1) the monomer, which packs into several (2) protofilaments that in turn associate to form a (3) fibril. Building upon our high resolution structure of the monomer of an amyloid-forming peptide from transthyretin (TTR105-115), we introduce single 1-13C labeled amino acids at seven different sites in the peptide and measure intermolecular carbonyl-carbonyl distances with an accuracy of ~0.11 A. Our results conclusively establish a parallel, in register, topology for the packing of this peptide into a β-sheet and provide constraints essential for the determination of an atomic resolution structure of the fibril. Furthermore, the approach we employ, based on a combination of a double-quantum filtered variant of the DRAWS recoupling sequence and multispin numerical simulations in SPINEVOLUTION, is general and should be applicable to a wide range of systems. PMID:20925357

  16. Accurate Determination of Rotational Energy Levels in the Ground State of ^{12}CH_4

    NASA Astrophysics Data System (ADS)

    Abe, M.; Iwakuni, K.; Okubo, S.; Sasada, H.

    2013-06-01

    We have measured absolute frequencies of saturated absorption of 183 allowed and 21 forbidden transitions in the νb{3} band of ^{12}CH_4 using an optical comb-referenced difference-frequency-generation spectrometer from 86.8 to 93.1 THz (from 2890 to 3100 wn). The pump and signal sources are a 1.06-μ m Nd:YAG laser and a 1.5-μ m extended-cavity laser diode. An enhanced-cavity absorption cell increases the optical electric field and enhances the sensitivity. The typical uncertainty is 3 kHz for the allowed transitions and 12 kHz for the forbidden transitions. Twenty combination differences are precisely determined, and the scalar rotational and centrifugal distortion constants of the ground state are thereby yielded as r@ = l@ r@ = l B_{{s}} (157 122 614.2 ± 1.5) kHz, D_{{s}} (3 328.545 ± 0.031) kHz, H_{{s}} (190.90 ± 0.26) Hz, and L_{{s}} (-13.16 ± 0.76) mHz. Here, B_{{s}} is the rotational constant and D_{{s}}, H_{{s}} and L_{{s}} are the scalar quartic, sextic, octic distortion constants. The relative uncertainties are considerably smaller than those obtained from global analysis of Fourier-transform infrared spectroscopy. S. Okubo, H. Nakayama, K. Iwakuni, H. Inaba and H. Sasada, Opt. Express 19, 23878 (2011). M. Abe, K. Iwakuni, S. Okubo, and H. Sasada, J. Opt. Soc. Am. B (to be published). S. Albert, S. Bauerecker, V. Boudon, L. R. Brown, J. -P. Champion, M. Loëte, A. Nikitin, and M. Quack, Chem. Phys. 356, 131 (2009).

  17. Genetic determinants of quantitative traits associated with cardiovascular disease risk.

    PubMed

    Smolková, Božena; Bonassi, Stefano; Buociková, Verona; Dušinská, Mária; Horská, Alexandra; Kuba, Daniel; Džupinková, Zuzana; Rašlová, Katarína; Gašparovič, Juraj; Slíž, Ivan; Ceppi, Marcello; Vohnout, Branislav; Wsólová, Ladislava; Volkovová, Katarína

    2015-08-01

    Established risk factors for cardiovascular diseases (CVD) may be moderated by genetic variants. In 2403 unrelated individuals from general practice (mean age 40.5 years), we evaluated the influence of 15 variants in 12 candidate genes on quantitative traits (QT) associated with CVD (body mass index, abdominal obesity, glucose, serum lipids, and blood pressure). Prior to multiple testing correction, univariate analysis associated APOE rs429358, rs7412 and ATG16L1 rs2241880 variants with serum lipid levels, while LEPR rs1137100 and ATG16L1 rs2241880 variants were linked to obesity related QTs. After taking into account confounding factors and correcting for multiple comparisons only APOE rs429358 and rs7412 variants remained significantly associated with risk of dyslipidemia. APOE rs429358 variant almost tripled the risk in homozygous subjects (OR = 2.97; 95% CI 1.09-8.10, p < 0.03) and had a lesser but still highly significant association also in heterozygous individuals (OR = 1.67; 95% CI 1.24-2.10; p < 0.001). Associations with hypertension, diabetes mellitus, and metabolic syndrome were not significant after Bonferroni correction. The influence of genetic variation is more evident in dyslipidemia than in other analyzed QTs. These results may contribute to strategic research aimed at including genetic variation in the set of data required to identify subjects at high risk of CVD. PMID:26043189

  18. Validation of quantitative magnetic resonance (QMR) for determination of body composition in rats.

    PubMed

    Johnson, M S; Smith, D L; Nagy, T R

    2009-09-01

    OBJECTIVE: To validate the use of quantitative magnetic resonance (QMR) to measure fat and lean mass in conscious rats. METHODS: Fifty Osborne-Mendel rats (249-770 g) were scanned using the Echo Medical 2 MHz body composition analyzer. Each rat was scanned under six settings (three acquisition times, with and without determination of total water). Precision was determined by the calculated coefficient of variation (CV) of three consecutive scans. Accuracy was determined by comparing the first scan to chemical carcass analysis and analyzed by paired t-tests and least-squares regression analyses. Twenty-five rats were used in the validation study, and 25 in the cross-validation study. RESULTS: The precision for fat, lean and water at all settings was <1%. QMR significantly overestimated fat (~5%; P<0.0001), and underestimated both lean (~12.5%; P<0.0001) and total water (~5.5%; P<0.0001). All QMR measures were significantly correlated with carcass measures (r(2)>0.99; P<0.0001). Using prediction equations from the validation study with the cross-validation rats, there were no significant differences between QMR fat and carcass fat at any setting (P>0.400). For four of the six QMR settings, there were no significant differences between QMR and carcass lean (P>0.05). For total water, all QMR settings were significantly different than carcass (P<0.05), but only by ~1%. CONCLUSIONS: QMR showed excellent precision for the determination of fat, lean and water. Despite overestimating fat and underestimating lean and water, all were highly related to carcass values. When tested in the cross-validation group, QMR fat could be accurately predicted at all settings; however, lean mass (two settings) and water were still slightly different (less than 1%). PMID:20686636

  19. Accurate determination of the temperature dependent thermalization coefficient (Q) in InAs/AlAsSb quantum wells

    NASA Astrophysics Data System (ADS)

    Esmaielpour, Hamidreza; Tang, Jinfeng; Whiteside, Vincent R.; Vijeyaragunathan, Sangeetha; Mishima, Tetsuya D.; Santos, Michael B.; Sellers, Ian R.

    2015-03-01

    We present an investigation of hot carriers in InAs/AlAsSb quantum wells as a practical candidate for a hot carrier solar cell absorber. The thermalization coefficient (Q) of the sample is investigated using continuous wave photoluminescence (PL). The Q is accurately determined through transfer matrix calculations of the absorption, analysis of the power density, penetration depth, diffusion, and recombination rates using a combination of simulation and empirical methods. A precise measurement of laser spot size is important in order to determine the absorbed power density. Simulations were performed based on our PL geometry in order to calculate the excitation spot size, which was compared with experiment by measurements using variable diameter pinholes to determine beam radius. Here, these techniques are described, in addition to, the temperature dependent hot carrier dynamics and phonon mediated thermalization coefficient for the InAs/AlAsSb quantum well structure.

  20. Accurate quantification of two key time points used in the determination of hydroxyl polyaluminum species by ferron timed spectrophotometry.

    PubMed

    Zhang, Jing; Yong, Xiaojing; Zhao, Dongyan; Shi, Qiuyi

    2015-01-01

    The content of mononuclear Al (Ala%) changed with its determination time (ta) under different dosages of Ferron (7-iodo-8-hydroxyquinoline-5-sulfonic acid, [Ferron]), and the change of Ala% with [Ferron] at different ta was systematically investigated for the first time. Thus, the most appropriate ta was found with the optimal [Ferron]. Also, the judgment of the platform (flat or level portion) of the complete reaction on the absorption-time curve determined in the hydroxyl polyaluminum solution by Ferron timed spectrophotometry (Ferron assay) was first digitized. The time point (tb) of complete reaction between the medium polyaluminum (Alb) and Ferron reagent depended on the reaction extent, and time could not be used only to judge. Thus, the tb was accurately determined and reduced to half of original, which improved the experiment efficiency significantly. The Ferron assay was completely optimized.

  1. Quantitative determination of 2'-deoxycytidine-5'-triphosphate in cell extracts by radioimmunoassay

    SciTech Connect

    Piall, E.M.; Aherne, G.W.; Marks, V.

    1986-04-01

    A radioimmunoassay (RIA) capable of quantitating dCTP in femtomolar amounts in cell extracts has been developed, and applied to human fibroblast cells lines and L5178Y mouse lymphoma lines. Cross reactivity of the antibody with CTP, though low (2.7%) has necessitated pre-RIA removal of CTP by either boronate affinity gel chromatography or sodium periodate oxidation. Fractions from the boronate gel column or aliquots of NaIO/sub 4/-treated cell extract are quantitated directly by the RIA. Recovery of extracted dCTP standard taken through the entire procedure is quantitative and results are reproductive. Due to the high sensitivity of the quantitation step, dCTP can be accurately measured in relatively small numbers of cells-about 10/sup 4/ cells.

  2. Multiclass semi-volatile compounds determination in wine by gas chromatography accurate time-of-flight mass spectrometry.

    PubMed

    Rodríguez-Cabo, T; Rodríguez, I; Ramil, M; Silva, A; Cela, R

    2016-04-15

    The performance of gas chromatography (GC) with accurate, high resolution mass spectrometry (HRMS) for the determination of a group of 39 semi-volatile compounds related to wine quality (pesticide residues, phenolic off-flavours, phenolic pollutants and bioactive stilbenes) is investigated. Solid-phase extraction (SPE) was used as extraction technique, previously to acetylation (phenolic compounds) and dispersive liquid-liquid microextraction (DLLME) concentration. Compounds were determined by GC coupled to a quadrupole time-of-flight (QTOF) MS system through an electron ionization (EI) source. The final method attained limits of quantification (LOQs) at the very low ng mL(-1) level, covering the range of expected concentrations for target compounds in red and white wines. For 38 out of 39 compounds, performance of sample preparation and determination steps were hardly affected by the wine matrix; thus, accurate recoveries were achieved by using pseudo-external calibration. Levels of target compounds in a set of 25 wine samples are reported. The capabilities of the described approach for the post-run identification of species not considered during method development, without retention time information, are illustrated and discussed with selected examples of compounds from different classes.

  3. Fast and accurate determination of 3D temperature distribution using fraction-step semi-implicit method

    NASA Astrophysics Data System (ADS)

    Cen, Wei; Hoppe, Ralph; Gu, Ning

    2016-09-01

    In this paper, we proposed a method to numerically determinate 3-dimensional thermal response due to electromagnetic exposure quickly and accurately. Due to the stability criterion the explicit finite-difference time-domain (FDTD) method works fast only if the spatial step is not set very small. In this paper, the semi-implicit Crank-Nicholson method for time domain discretization with unconditional time stability is proposed, where the idea of fractional steps method was utilized in 3-dimension so that an efficient numerical implementation is obtained. Compared with the explicit FDTD, with similar numerical precision, the proposed method takes less than 1/200 of the execution time.

  4. Note: Accurate determination of thickness of multiple layers of thin film deposited on a piezoelectric quartz crystal.

    PubMed

    Wajid, Abdul

    2013-10-01

    Modern day piezoelectric quartz crystal microbalances for thin film deposition control are based on Z-match equation, which is mathematically valid for deposition of a single material on a given quartz crystal. When multiple layers are deposited, thickness and deposition rate errors accumulate due to mismatch of acoustic impedance of different materials. Here we present a novel method, based on the acoustic transfer matrix formalism, for accurate determination of thickness of an arbitrary number of layers of dissimilar materials deposited on a quartz crystal. Laboratory data show excellent accuracy of the method compared to conventional Z-match equation. PMID:24182174

  5. Quantitative Determination of Citric and Ascorbic Acid in Powdered Drink Mixes

    ERIC Educational Resources Information Center

    Sigmann, Samuella B.; Wheeler, Dale E.

    2004-01-01

    A procedure by which the reactions are used to quantitatively determine the amount of total acid, the amount of total ascorbic acid and the amount of citric acid in a given sample of powdered drink mix, are described. A safe, reliable and low-cost quantitative method to analyze consumer product for acid content is provided.

  6. Quantitative determination of steroid acetates in pharmaceutical preparations by high performance liquid chromatography.

    PubMed

    van Dame, H C

    1980-11-01

    A high performance liquid chromatographic method is described for the rapid, quantitative determination of corticosteroid acetates in a variety of pharmaceutical preparations. The method not only separates the steroids from other ingredients in tablets, creams, ointments, lotions, or suspensions, but also from their probable degradation products. The steroid acetate is extracted from a tablet matrix or from a suspension with water and acetonitrile (50 + 50), or from a hexane suspension of a cream, ointment, or lotion with acetonitrile. A liquid chromatograph equipped with an accurate flow controller, a variable wavelength detector capable of measurement at 240 nm, and a 10 microL loop injector was used. Components were separated on a microBondpak C18 column with acetonitrile-water as the mobile phase. K' values of 10 commonly used steroid acetates are shown and compared with Rf values from thin layer chromatographic systems. Results were equivalent for cortisone acetate tablets assayed by both the proposed and the official USP methods. Other pharmaceutical preparations were assayed to test the method, with satisfactory results.

  7. Quantitative capillary electrophoresis determination of oversulfated chondroitin sulfate as a contaminant in heparin preparations

    PubMed Central

    Volpi, Nicola; Maccari, Francesca; Linhardt, Robert J.

    2014-01-01

    A simple, accurate and robust quantitative CE method for the determination of oversulfated chondroitin sulfate (OSCS) as a contaminant in heparin (Hep) preparations is described. After degradation of the polysaccharides by acidic hydrolysis, the hexosamines produced, i.e., GlcN from Hep and GalN from OSCS, were derivatized with anthranilic acid (AA) and separated by means of CE in approx. 10 min with high sensitivity detection at 214 nm (limit of detection (LOD) of approx. 200 pg). Furthermore, AA-derivatized GlcN and GalN showed quite similar molar absorptivity allowing for direct and simple quantification of OSCS in Hep samples. Moreover, a preliminary step of specific enzymatic treatment by using chondroitin ABC lyase may be applied for the specific elimination of interference in the analysis due to the possible presence in Hep samples of natural chondroitin sulfate and dermatan sulfate impurities, making this analytical approach highly specific for OSCS contamination, since chondroitin ABC lyase is unable to act on this semi-synthetic polymer. The CE method was validated for specificity, linearity, accuracy, precision, LOD and limit of quantification (LOQ). Due to the very high sensitivity of CE, as little as 1% OSCS contaminant in Hep sample could be detected and quantified. Finally, a contaminated raw Hep sample was found to contain 38.9% OSCS while a formulated contaminated Hep was calculated to have 39.7% OSCS. PMID:19232311

  8. A field instrument for quantitative determination of beryllium by activation analysis

    USGS Publications Warehouse

    Vaughn, William W.; Wilson, E.E.; Ohm, J.M.

    1960-01-01

    A low-cost instrument has been developed for quantitative determinations of beryllium in the field by activation analysis. The instrument makes use of the gamma-neutron reaction between gammas emitted by an artificially radioactive source (Sb124) and beryllium as it occurs in nature. The instrument and power source are mounted in a panel-type vehicle. Samples are prepared by hand-crushing the rock to approximately ?-inch mesh size and smaller. Sample volumes are kept constant by means of a standard measuring cup. Instrument calibration, made by using standards of known BeO content, indicates the analyses are reproducible and accurate to within ? 0.25 percent BeO in the range from 1 to 20 percent BeO with a sample counting time of 5 minutes. Sensitivity of the instrument maybe increased somewhat by increasing the source size, the sample size, or by enlarging the cross-sectional area of the neutron-sensitive phosphor normal to the neutron flux.

  9. Simultaneous measurement in mass and mass/mass mode for accurate qualitative and quantitative screening analysis of pharmaceuticals in river water.

    PubMed

    Martínez Bueno, M J; Ulaszewska, Maria M; Gomez, M J; Hernando, M D; Fernández-Alba, A R

    2012-09-21

    A new approach for the analysis of pharmaceuticals (target and non-target) in water by LC-QTOF-MS is described in this work. The study has been designed to assess the performance of the simultaneous quantitative screening of target compounds, and the qualitative analysis of non-target analytes, in just one run. The features of accurate mass full scan mass spectrometry together with high MS/MS spectral acquisition rates - by means of information dependent acquisition (IDA) - have demonstrated their potential application in this work. Applying this analytical strategy, an identification procedure is presented based on library searching for compounds which were not included a priori in the analytical method as target compounds, thus allowing their characterization by data processing of accurate mass measurements in MS and MS/MS mode. The non-target compounds identified in river water samples were ketorolac, trazodone, fluconazole, metformin and venlafaxine. Simultaneously, this strategy allowed for the identification of other compounds which were not included in the library by screening the highest intensity peaks detected in the samples and by analysis of the full scan TOF-MS, isotope pattern and MS/MS spectra - the example of loratadine (histaminergic) is described. The group of drugs of abuse selected as target compounds for evaluation included analgesics, opioids and psychostimulants. Satisfactory results regarding sensitivity and linearity of the developed method were obtained. Limits of detection for the selected target compounds were from 0.003 to 0.01 μg/L and 0.01 to 0.5 μg/L, in MS and MS/MS mode, respectively - by direct sample injection of 100 μL.

  10. Accurate, quantitative assays for the hydrolysis of soluble type I, II, and III /sup 3/H-acetylated collagens by bacterial and tissue collagenases

    SciTech Connect

    Mallya, S.K.; Mookhtiar, K.A.; Van Wart, H.E.

    1986-11-01

    Accurate and quantitative assays for the hydrolysis of soluble /sup 3/H-acetylated rat tendon type I, bovine cartilage type II, and human amnion type III collagens by both bacterial and tissue collagenases have been developed. The assays are carried out at any temperature in the 1-30/sup 0/C range in a single reaction tube and the progress of the reaction is monitored by withdrawing aliquots as a function of time, quenching with 1,10-phenanthroline, and quantitation of the concentration of hydrolysis fragments. The latter is achieved by selective denaturation of these fragments by incubation under conditions described in the previous paper of this issue. The assays give percentages of hydrolysis of all three collagen types by neutrophil collagenase that agree well with the results of gel electrophoresis experiments. The initial rates of hydrolysis of all three collagens are proportional to the concentration of both neutrophil or Clostridial collagenases over a 10-fold range of enzyme concentrations. All three assays can be carried out at collagen concentrations that range from 0.06 to 2 mg/ml and give linear double reciprocal plots for both tissue and bacterial collagenases that can be used to evaluate the kinetic parameters K/sub m/ and k/sub cat/ or V/sub max/. The assay developed for the hydrolysis of rat type I collagen by neutrophil collagenase is shown to be more sensitive by at least one order of magnitude than comparable assays that use rat type I collagen fibrils or gels as substrate.

  11. Quantitative determination of fucoidan using polyion-sensitive membrane electrodes.

    PubMed

    Kim, Ji Min; Nguyen, Loc; Barr, Mary Frances; Morabito, Michael; Stringer, Damien; Fitton, J Helen; Mowery, Kelly A

    2015-06-01

    The use of polyanion and polycation-sensitive membrane electrodes to detect five different preparations of fucoidan is described. Unlike linear polyanionic molecules previously measured with polymer membrane-based electrochemical sensors, fucoidans from marine brown algae are all highly branched, sulfated polysaccharides with varying charge densities and structures, depending on the species of seaweed, method of extraction used and extent of purification. When tridodecylmethylammonium (TDMA) was used as the ion-exchanger, a large, non-equilibrium EMF response was observed over a concentration range of 0.5-50 μg mL(-1) fucoidan. Fucoidan was also measured by titration with polycationic protamine, using a dinonylnaphthalene sulfonate (DNNS)-doped membrane electrode as the potentiometric endpoint detector. Potentiometric titration was used to determine the binding ratio between protamine and fucoidan at the neutralization endpoint for each fucoidan preparation. This binding ratio was then used to successfully determine the fucoidan content of commercially available nutritional supplements. Fucoidan was also measured in undiluted blood serum, demonstrating that this method may be applicable for measuring fucoidan for clinical applications.

  12. Qualitative and quantitative determination of ubiquinones by the method of high-efficiency liquid chromatography

    SciTech Connect

    Yanotovskii, M.T.; Mogilevskaya, M.P.; Obol'nikova, E.A.; Kogan, L.M.; Samokhvalov, G.I.

    1986-07-10

    A method has been developed for the qualitative and quantitative determination of ubiquinones CoQ/sub 6/-CoQ/sub 10/, using high-efficiency reversed-phase liquid chromatography. Tocopherol acetate was used as the internal standard.

  13. A rapid and accurate method for the quantitative estimation of natural polysaccharides and their fractions using high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector.

    PubMed

    Cheong, Kit-Leong; Wu, Ding-Tao; Zhao, Jing; Li, Shao-Ping

    2015-06-26

    In this study, a rapid and accurate method for quantitative analysis of natural polysaccharides and their different fractions was developed. Firstly, high performance size exclusion chromatography (HPSEC) was utilized to separate natural polysaccharides. And then the molecular masses of their fractions were determined by multi-angle laser light scattering (MALLS). Finally, quantification of polysaccharides or their fractions was performed based on their response to refractive index detector (RID) and their universal refractive index increment (dn/dc). Accuracy of the developed method for the quantification of individual and mixed polysaccharide standards, including konjac glucomannan, CM-arabinan, xyloglucan, larch arabinogalactan, oat β-glucan, dextran (410, 270, and 25 kDa), mixed xyloglucan and CM-arabinan, and mixed dextran 270 K and CM-arabinan was determined, and their average recoveries were between 90.6% and 98.3%. The limits of detection (LOD) and quantification (LOQ) were ranging from 10.68 to 20.25 μg/mL, and 42.70 to 68.85 μg/mL, respectively. Comparing to the conventional phenol sulfuric acid assay and HPSEC coupled with evaporative light scattering detection (HPSEC-ELSD) analysis, the developed HPSEC-MALLS-RID method based on universal dn/dc for the quantification of polysaccharides and their fractions is much more simple, rapid, and accurate with no need of individual polysaccharide standard, as well as free of calibration curve. The developed method was also successfully utilized for quantitative analysis of polysaccharides and their different fractions from three medicinal plants of Panax genus, Panax ginseng, Panax notoginseng and Panax quinquefolius. The results suggested that the HPSEC-MALLS-RID method based on universal dn/dc could be used as a routine technique for the quantification of polysaccharides and their fractions in natural resources.

  14. A rapid and accurate method for the quantitative estimation of natural polysaccharides and their fractions using high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector.

    PubMed

    Cheong, Kit-Leong; Wu, Ding-Tao; Zhao, Jing; Li, Shao-Ping

    2015-06-26

    In this study, a rapid and accurate method for quantitative analysis of natural polysaccharides and their different fractions was developed. Firstly, high performance size exclusion chromatography (HPSEC) was utilized to separate natural polysaccharides. And then the molecular masses of their fractions were determined by multi-angle laser light scattering (MALLS). Finally, quantification of polysaccharides or their fractions was performed based on their response to refractive index detector (RID) and their universal refractive index increment (dn/dc). Accuracy of the developed method for the quantification of individual and mixed polysaccharide standards, including konjac glucomannan, CM-arabinan, xyloglucan, larch arabinogalactan, oat β-glucan, dextran (410, 270, and 25 kDa), mixed xyloglucan and CM-arabinan, and mixed dextran 270 K and CM-arabinan was determined, and their average recoveries were between 90.6% and 98.3%. The limits of detection (LOD) and quantification (LOQ) were ranging from 10.68 to 20.25 μg/mL, and 42.70 to 68.85 μg/mL, respectively. Comparing to the conventional phenol sulfuric acid assay and HPSEC coupled with evaporative light scattering detection (HPSEC-ELSD) analysis, the developed HPSEC-MALLS-RID method based on universal dn/dc for the quantification of polysaccharides and their fractions is much more simple, rapid, and accurate with no need of individual polysaccharide standard, as well as free of calibration curve. The developed method was also successfully utilized for quantitative analysis of polysaccharides and their different fractions from three medicinal plants of Panax genus, Panax ginseng, Panax notoginseng and Panax quinquefolius. The results suggested that the HPSEC-MALLS-RID method based on universal dn/dc could be used as a routine technique for the quantification of polysaccharides and their fractions in natural resources. PMID:25990349

  15. Quantitative determination of activation energies in mechanochemical reactions.

    PubMed

    Fischer, Franziska; Wenzel, Klaus-Jürgen; Rademann, Klaus; Emmerling, Franziska

    2016-08-17

    Mechanochemical reactions often result in 100% yields of single products, making purifying procedures obsolete. Mechanochemistry is also a sustainable and eco-friendly method. The ever increasing interest in this method is contrasted by a lack in mechanistic understanding of the mechanochemical reactivity and selectivity. Recent in situ investigations provided direct insight into formation pathways. However, the currently available theories do not predict temperature T as an influential factor. Here, we report the first determination of an apparent activation energy for a mechanochemical reaction. In a temperature-dependent in situ study the cocrystallisation of ibuprofen and nicotinamide was investigated as a model system. These experiments provide a pivotal step towards a comprehensive understanding of milling reaction mechanisms.

  16. Quantitative Interpretation of Tracks for Determination of Body Mass

    PubMed Central

    Schanz, Tom; Lins, Yvonne; Viefhaus, Hanna; Barciaga, Thomas; Läbe, Sashima; Preuschoft, Holger; Witzel, Ulrich; Sander, P. Martin

    2013-01-01

    To better understand the biology of extinct animals, experimentation with extant animals and innovative numerical approaches have grown in recent years. This research project uses principles of soil mechanics and a neoichnological field experiment with an African elephant to derive a novel concept for calculating the mass (i.e., the weight) of an animal from its footprints. We used the elephant's footprint geometry (i.e., vertical displacements, diameter) in combination with soil mechanical analyses (i.e., soil classification, soil parameter determination in the laboratory, Finite Element Analysis (FEA) and gait analysis) for the back analysis of the elephant's weight from a single footprint. In doing so we validated the first component of a methodology for calculating the weight of extinct dinosaurs. The field experiment was conducted under known boundary conditions at the Zoological Gardens Wuppertal with a female African elephant. The weight of the elephant was measured and the walking area was prepared with sediment in advance. Then the elephant was walked across the test area, leaving a trackway behind. Footprint geometry was obtained by laser scanning. To estimate the dynamic component involved in footprint formation, the velocity the foot reaches when touching the subsoil was determined by the Digital Image Correlation (DIC) technique. Soil parameters were identified by performing experiments on the soil in the laboratory. FEA was then used for the backcalculation of the elephant's weight. With this study, we demonstrate the adaptability of using footprint geometry in combination with theoretical considerations of loading of the subsoil during a walk and soil mechanical methods for prediction of trackmakers weight. PMID:24204890

  17. Quantitative interpretation of tracks for determination of body mass.

    PubMed

    Schanz, Tom; Lins, Yvonne; Viefhaus, Hanna; Barciaga, Thomas; Läbe, Sashima; Preuschoft, Holger; Witzel, Ulrich; Sander, P Martin

    2013-01-01

    To better understand the biology of extinct animals, experimentation with extant animals and innovative numerical approaches have grown in recent years. This research project uses principles of soil mechanics and a neoichnological field experiment with an African elephant to derive a novel concept for calculating the mass (i.e., the weight) of an animal from its footprints. We used the elephant's footprint geometry (i.e., vertical displacements, diameter) in combination with soil mechanical analyses (i.e., soil classification, soil parameter determination in the laboratory, Finite Element Analysis (FEA) and gait analysis) for the back analysis of the elephant's weight from a single footprint. In doing so we validated the first component of a methodology for calculating the weight of extinct dinosaurs. The field experiment was conducted under known boundary conditions at the Zoological Gardens Wuppertal with a female African elephant. The weight of the elephant was measured and the walking area was prepared with sediment in advance. Then the elephant was walked across the test area, leaving a trackway behind. Footprint geometry was obtained by laser scanning. To estimate the dynamic component involved in footprint formation, the velocity the foot reaches when touching the subsoil was determined by the Digital Image Correlation (DIC) technique. Soil parameters were identified by performing experiments on the soil in the laboratory. FEA was then used for the backcalculation of the elephant's weight. With this study, we demonstrate the adaptability of using footprint geometry in combination with theoretical considerations of loading of the subsoil during a walk and soil mechanical methods for prediction of trackmakers weight. PMID:24204890

  18. Rapid and accurate determination of total lung capacity (TLC) from routine chest radiograms using a programmable hand-held calculator.

    PubMed

    Rodgers, R P; Tannen, R

    1983-01-01

    Since its appearance in 1960, the method of Barnhard and associates for the determination of total lung capacity (TLC) from routine chest radiograms has been widely studied in normal and diseased subjects. The method appears to be as accurate as the current definitive procedure, total body plethysmography. The method is in routine use in major medical institutions where the procedure has been automated, but the method does not seem to have gained the wide use it deserves. This is likely due to the tedium of the technique when performed manually--a single determination can require 30 min. We present here an implementation of the Barnhard method for the HP41-C hand-held programmable calculator. In conjunction with the use of a transparent reticle used for obtaining the required measurements, the program allows a single measurement to be made in under 12 minutes. We hope this technique will make radiographic TLC measurements more broadly accessible to the medical profession. PMID:6872526

  19. Quantitative determination of mineral composition by powder X-ray diffraction

    DOEpatents

    Pawloski, Gayle A.

    1986-01-01

    An external standard intensity ratio method is used for quantitatively determining mineralogic compositions of samples by x-ray diffraction. The method uses ratios of x-ray intensity peaks from a single run. Constants are previously determined for each mineral which is to be quantitatively measured. Ratios of the highest intensity peak of each mineral to be quantified in the sample and the highest intensity peak of a reference mineral contained in the sample are used to calculate sample composition.

  20. Quantitative determination of mineral composition by powder x-ray diffraction

    DOEpatents

    Pawloski, G.A.

    1984-08-10

    An external standard intensity ratio method is used for quantitatively determining mineralogic compositions of samples by x-ray diffraction. The method uses ratios of x-ray intensity peaks from a single run. Constants are previously determined for each mineral which is to be quantitatively measured. Ratios of the highest intensity peak of each mineral to be quantified in the sample and the highest intensity peak of a reference mineral contained in the sample are used to calculate sample composition.

  1. Two-dimensional flow nanometry of biological nanoparticles for accurate determination of their size and emission intensity

    PubMed Central

    Block, Stephan; Fast, Björn Johansson; Lundgren, Anders; Zhdanov, Vladimir P.; Höök, Fredrik

    2016-01-01

    Biological nanoparticles (BNPs) are of high interest due to their key role in various biological processes and use as biomarkers. BNP size and composition are decisive for their functions, but simultaneous determination of both properties with high accuracy remains challenging. Optical microscopy allows precise determination of fluorescence/scattering intensity, but not the size of individual BNPs. The latter is better determined by tracking their random motion in bulk, but the limited illumination volume for tracking this motion impedes reliable intensity determination. Here, we show that by attaching BNPs to a supported lipid bilayer, subjecting them to hydrodynamic flows and tracking their motion via surface-sensitive optical imaging enable determination of their diffusion coefficients and flow-induced drifts, from which accurate quantification of both BNP size and emission intensity can be made. For vesicles, the accuracy of this approach is demonstrated by resolving the expected radius-squared dependence of their fluorescence intensity for radii down to 15 nm. PMID:27658367

  2. Quantitative Determination of Isotope Ratios from Experimental Isotopic Distributions

    PubMed Central

    Kaur, Parminder; O’Connor, Peter B.

    2008-01-01

    Isotope variability due to natural processes provides important information for studying a variety of complex natural phenomena from the origins of a particular sample to the traces of biochemical reaction mechanisms. These measurements require high-precision determination of isotope ratios of a particular element involved. Isotope Ratio Mass Spectrometers (IRMS) are widely employed tools for such a high-precision analysis, which have some limitations. This work aims at overcoming the limitations inherent to IRMS by estimating the elemental isotopic abundance from the experimental isotopic distribution. In particular, a computational method has been derived which allows the calculation of 13C/12C ratios from the whole isotopic distributions, given certain caveats, and these calculations are applied to several cases to demonstrate their utility. The limitations of the method in terms of the required number of ions and S/N ratio are discussed. For high-precision estimates of the isotope ratios, this method requires very precise measurement of the experimental isotopic distribution abundances, free from any artifacts introduced by noise, sample heterogeneity, or other experimental sources. PMID:17263354

  3. Quantitative Determination of Lineshape Parameters from Velocity Modulation Spectroscopy

    NASA Astrophysics Data System (ADS)

    Hodges, James N.; McCall, Benjamin J.

    2016-06-01

    Velocity Modulation Spectroscopy (VMS) has stood as the gold standard in molecular ion spectroscopy for 30 years. Whether in a traditional uni-directional experiment or more complicated cavity-enhanced layouts with additional layers of modulation, VMS remains the preferred ion detection scheme and is responsible for the detection and transition frequency determination of around 50 molecules. Despite its success, VMS still has a great deal of untapped potential. There have only been two other published studies of VMS lineshapes and both struggle with the highly correlated parameters: linewidth, intensity, and velocity modulation amplitude, i.e. the maximum Doppler shift during a period of the discharge. Due to this difficulty, both Gao and Civis made concessions to achieve a good fit. Careful analysis of the contour of the transition profile allows us to properly disentangle those parameters in order to probe the environment of the positive column. We can extract the precise values for the translational temperature of the ion, the relative transition intensity, the ion mobility, and the electric field strength just from the lineshape of a single transition. A firm understanding of the lineshape will facilitate chemical and physical investigations of positive columns and allow for a better understanding of more complicated detection schemes. H. Gao et al., Acta Phys. Sin. 50, 1463 (2001) S. Civis, Chem. Phys. 186, 63 (1994)

  4. Allele Specific Locked Nucleic Acid Quantitative PCR (ASLNAqPCR): An Accurate and Cost-Effective Assay to Diagnose and Quantify KRAS and BRAF Mutation

    PubMed Central

    Morandi, Luca; de Biase, Dario; Visani, Michela; Cesari, Valentina; De Maglio, Giovanna; Pizzolitto, Stefano; Pession, Annalisa; Tallini, Giovanni

    2012-01-01

    The use of tyrosine kinase inhibitors (TKIs) requires the testing for hot spot mutations of the molecular effectors downstream the membrane-bound tyrosine kinases since their wild type status is expected for response to TKI therapy. We report a novel assay that we have called Allele Specific Locked Nucleic Acid quantitative PCR (ASLNAqPCR). The assay uses LNA-modified allele specific primers and LNA-modified beacon probes to increase sensitivity, specificity and to accurately quantify mutations. We designed primers specific for codon 12/13 KRAS mutations and BRAF V600E, and validated the assay with 300 routine samples from a variety of sources, including cytology specimens. All were analyzed by ASLNAqPCR and Sanger sequencing. Discordant cases were pyrosequenced. ASLNAqPCR correctly identified BRAF and KRAS mutations in all discordant cases and all had a mutated/wild type DNA ratio below the analytical sensitivity of the Sanger method. ASLNAqPCR was 100% specific with greater accuracy, positive and negative predictive values compared with Sanger sequencing. The analytical sensitivity of ASLNAqPCR is 0.1%, allowing quantification of mutated DNA in small neoplastic cell clones. ASLNAqPCR can be performed in any laboratory with real-time PCR equipment, is very cost-effective and can easily be adapted to detect hot spot mutations in other oncogenes. PMID:22558339

  5. Allele specific locked nucleic acid quantitative PCR (ASLNAqPCR): an accurate and cost-effective assay to diagnose and quantify KRAS and BRAF mutation.

    PubMed

    Morandi, Luca; de Biase, Dario; Visani, Michela; Cesari, Valentina; De Maglio, Giovanna; Pizzolitto, Stefano; Pession, Annalisa; Tallini, Giovanni

    2012-01-01

    The use of tyrosine kinase inhibitors (TKIs) requires the testing for hot spot mutations of the molecular effectors downstream the membrane-bound tyrosine kinases since their wild type status is expected for response to TKI therapy. We report a novel assay that we have called Allele Specific Locked Nucleic Acid quantitative PCR (ASLNAqPCR). The assay uses LNA-modified allele specific primers and LNA-modified beacon probes to increase sensitivity, specificity and to accurately quantify mutations. We designed primers specific for codon 12/13 KRAS mutations and BRAF V600E, and validated the assay with 300 routine samples from a variety of sources, including cytology specimens. All were analyzed by ASLNAqPCR and Sanger sequencing. Discordant cases were pyrosequenced. ASLNAqPCR correctly identified BRAF and KRAS mutations in all discordant cases and all had a mutated/wild type DNA ratio below the analytical sensitivity of the Sanger method. ASLNAqPCR was 100% specific with greater accuracy, positive and negative predictive values compared with Sanger sequencing. The analytical sensitivity of ASLNAqPCR is 0.1%, allowing quantification of mutated DNA in small neoplastic cell clones. ASLNAqPCR can be performed in any laboratory with real-time PCR equipment, is very cost-effective and can easily be adapted to detect hot spot mutations in other oncogenes.

  6. Quantitative determination of cesium binding to ferric hexacyanoferrate: Prussian blue.

    PubMed

    Faustino, Patrick J; Yang, Yongsheng; Progar, Joseph J; Brownell, Charles R; Sadrieh, Nakissa; May, Joan C; Leutzinger, Eldon; Place, David A; Duffy, Eric P; Houn, Florence; Loewke, Sally A; Mecozzi, Vincent J; Ellison, Christopher D; Khan, Mansoor A; Hussain, Ajaz S; Lyon, Robbe C

    2008-05-12

    Ferric hexacyanoferrate (Fe4III[FeII(CN)6]3), also known as insoluble Prussian blue (PB) is the active pharmaceutical ingredient (API) of the drug product, Radiogardase. Radiogardase is the first FDA approved medical countermeasure for the treatment of internal contamination with radioactive cesium (Cs) or thallium in the event of a major radiological incident such as a "dirty bomb". A number of pre-clinical and clinical studies have evaluated the use of PB as an investigational decorporation agent to enhance the excretion of metal cations. There are few sources of published in vitro data that detail the binding capacity of cesium to insoluble PB under various chemical and physical conditions. The study objective was to determine the in vitro binding capacity of PB APIs and drug products by evaluating certain chemical and physical factors such as medium pH, particle size, and storage conditions (temperature). In vitro experimental conditions ranged from pH 1 to 9, to cover the range of pH levels that PB may encounter in the gastrointestinal (GI) tract in humans. Measurements of cesium binding were made between 1 and 24h, to cover gastric and intestinal tract residence time using a validated atomic emission spectroscopy (AES) method. The results indicated that pH, exposure time, storage temperature (affecting moisture content) and particle size play significant roles in the cesium binding to both the PB API and the drug product. The lowest cesium binding was observed at gastric pH of 1 and 2, whereas the highest cesium binding was observed at physiological pH of 7.5. It was observed that dry storage conditions resulted in a loss of moisture from PB, which had a significant negative effect on the PB cesium binding capacity at time intervals consistent with gastric residence. Differences were also observed in the binding capacity of PB with different particle sizes. Significant batch to batch differences were also observed in the binding capacity of some PB API and

  7. Quantitative determination of cesium binding to ferric hexacyanoferrate: Prussian blue.

    PubMed

    Faustino, Patrick J; Yang, Yongsheng; Progar, Joseph J; Brownell, Charles R; Sadrieh, Nakissa; May, Joan C; Leutzinger, Eldon; Place, David A; Duffy, Eric P; Houn, Florence; Loewke, Sally A; Mecozzi, Vincent J; Ellison, Christopher D; Khan, Mansoor A; Hussain, Ajaz S; Lyon, Robbe C

    2008-05-12

    Ferric hexacyanoferrate (Fe4III[FeII(CN)6]3), also known as insoluble Prussian blue (PB) is the active pharmaceutical ingredient (API) of the drug product, Radiogardase. Radiogardase is the first FDA approved medical countermeasure for the treatment of internal contamination with radioactive cesium (Cs) or thallium in the event of a major radiological incident such as a "dirty bomb". A number of pre-clinical and clinical studies have evaluated the use of PB as an investigational decorporation agent to enhance the excretion of metal cations. There are few sources of published in vitro data that detail the binding capacity of cesium to insoluble PB under various chemical and physical conditions. The study objective was to determine the in vitro binding capacity of PB APIs and drug products by evaluating certain chemical and physical factors such as medium pH, particle size, and storage conditions (temperature). In vitro experimental conditions ranged from pH 1 to 9, to cover the range of pH levels that PB may encounter in the gastrointestinal (GI) tract in humans. Measurements of cesium binding were made between 1 and 24h, to cover gastric and intestinal tract residence time using a validated atomic emission spectroscopy (AES) method. The results indicated that pH, exposure time, storage temperature (affecting moisture content) and particle size play significant roles in the cesium binding to both the PB API and the drug product. The lowest cesium binding was observed at gastric pH of 1 and 2, whereas the highest cesium binding was observed at physiological pH of 7.5. It was observed that dry storage conditions resulted in a loss of moisture from PB, which had a significant negative effect on the PB cesium binding capacity at time intervals consistent with gastric residence. Differences were also observed in the binding capacity of PB with different particle sizes. Significant batch to batch differences were also observed in the binding capacity of some PB API and

  8. Accurate retention time determination of co-eluting proteins in analytical chromatography by means of spectral data.

    PubMed

    Dismer, Florian; Hansen, Sigrid; Oelmeier, Stefan Alexander; Hubbuch, Jürgen

    2013-03-01

    Chromatography is the method of choice for the separation of proteins, at both analytical and preparative scale. Orthogonal purification strategies for industrial use can easily be implemented by combining different modes of adsorption. Nevertheless, with flexibility comes the freedom of choice and optimal conditions for consecutive steps need to be identified in a robust and reproducible fashion. One way to address this issue is the use of mathematical models that allow for an in silico process optimization. Although this has been shown to work, model parameter estimation for complex feedstocks becomes the bottleneck in process development. An integral part of parameter assessment is the accurate measurement of retention times in a series of isocratic or gradient elution experiments. As high-resolution analytics that can differentiate between proteins are often not readily available, pure protein is mandatory for parameter determination. In this work, we present an approach that has the potential to solve this problem. Based on the uniqueness of UV absorption spectra of proteins, we were able to accurately measure retention times in systems of up to four co-eluting compounds. The presented approach is calibration-free, meaning that prior knowledge of pure component absorption spectra is not required. Actually, pure protein spectra can be determined from co-eluting proteins as part of the methodology. The approach was tested for size-exclusion chromatograms of 38 mixtures of co-eluting proteins. Retention times were determined with an average error of 0.6 s (1.6% of average peak width), approximated and measured pure component spectra showed an average coefficient of correlation of 0.992.

  9. THE EFFECT OF STARSPOTS ON ACCURATE RADIUS DETERMINATION OF THE LOW-MASS DOUBLE-LINED ECLIPSING BINARY GU Boo

    SciTech Connect

    Windmiller, G.; Orosz, J. A.; Etzel, P. B. E-mail: orosz@sciences.sdsu.ed

    2010-04-01

    GU Boo is one of only a relatively small number of well-studied double-lined eclipsing binaries that contain low-mass stars. Lopez-Morales and Ribas present a comprehensive analysis of multi-color light and radial velocity curves for this system. The GU Boo light curves presented by Lopez-Morales and Ribas had substantial asymmetries, which were attributed to large spots. In spite of the asymmetry, Lopez-Morales and Ribas derived masses and radii accurate to {approx_equal}2%. We obtained additional photometry of GU Boo using both a CCD and a single-channel photometer and modeled the light curves with the ELC software to determine if the large spots in the light curves give rise to systematic errors at the few percent level. We also modeled the original light curves from the work of Lopez-Morales and Ribas using models with and without spots. We derived a radius of the primary of 0.6329 +- 0.0026 R{sub sun}, 0.6413 +- 0.0049 R{sub sun}, and 0.6373 +- 0.0029 R{sub sun} from the CCD, photoelectric, and Lopez-Morales and Ribas data, respectively. Each of these measurements agrees with the value reported by Lopez-Morales and Ribas (R{sub 1} = 0.623 +- 0.016 R{sub sun}) at the level of {approx}2%. In addition, the spread in these values is {approx}1%-2% from the mean. For the secondary, we derive radii of 0.6074 +- 0.0035 R{sub sun}, 0.5944 +- 0.0069 R{sub sun}, and 0.5976 +- 0.0059 R{sub sun} from the three respective data sets. The Lopez-Morales and Ribas value is R{sub 2} = 0.620 +- 0.020 R{sub sun}, which is {approx}2%-3% larger than each of the three values we found. The spread in these values is {approx}2% from the mean. The systematic difference between our three determinations of the secondary radius and that of Lopez-Morales and Ribas might be attributed to differences in the modeling process and codes used. Our own fits suggest that, for GU Boo at least, using accurate spot modeling of a single set of multi-color light curves results in radii determinations

  10. Accurate determination of the valence band edge in hard x-ray photoemission spectra using GW theory

    NASA Astrophysics Data System (ADS)

    Lischner, Johannes; Nemšák, Slavomír; Conti, Giuseppina; Gloskovskii, Andrei; Pálsson, Gunnar Karl; Schneider, Claus M.; Drube, Wolfgang; Louie, Steven G.; Fadley, Charles

    2016-04-01

    We introduce a new method for determining accurate values of the valence-band maximum in x-ray photoemission spectra. Specifically, we align the sharpest peak in the valence-band region of the experimental spectrum with the corresponding feature of a theoretical valence-band density of states curve from ab initio GW theory calculations. This method is particularly useful for soft and hard x-ray photoemission studies of materials with a mixture of valence-band characters, where strong matrix element effects can render standard methods for extracting the valence-band maximum unreliable. We apply our method to hydrogen-terminated boron-doped diamond, which is a promising substrate material for novel solar cell devices. By carrying out photoemission experiments with variable light polarizations, we verify the accuracy of our analysis and the general validity of the method.

  11. Accurate determination of genetic identity for a single cacao bean, using molecular markers with a nanofluidic system, ensures cocoa authentication.

    PubMed

    Fang, Wanping; Meinhardt, Lyndel W; Mischke, Sue; Bellato, Cláudia M; Motilal, Lambert; Zhang, Dapeng

    2014-01-15

    Cacao (Theobroma cacao L.), the source of cocoa, is an economically important tropical crop. One problem with the premium cacao market is contamination with off-types adulterating raw premium material. Accurate determination of the genetic identity of single cacao beans is essential for ensuring cocoa authentication. Using nanofluidic single nucleotide polymorphism (SNP) genotyping with 48 SNP markers, we generated SNP fingerprints for small quantities of DNA extracted from the seed coat of single cacao beans. On the basis of the SNP profiles, we identified an assumed adulterant variety, which was unambiguously distinguished from the authentic beans by multilocus matching. Assignment tests based on both Bayesian clustering analysis and allele frequency clearly separated all 30 authentic samples from the non-authentic samples. Distance-based principle coordinate analysis further supported these results. The nanofluidic SNP protocol, together with forensic statistical tools, is sufficiently robust to establish authentication and to verify gourmet cacao varieties. This method shows significant potential for practical application.

  12. Accurate Determination of Radionuclidic Purity and Half-Life of Reactor Produced LU-177G for Metabolic Radioimmunotherapy

    NASA Astrophysics Data System (ADS)

    Groppi, F.; Canella, L.; Bonardi, M. L.; Zona, C.; Morzenti, S.; Menapace, E.; Alfassi, Z. B.; Chinol, M.; Papi, S.; Tosi, G.

    2006-04-01

    The accurate determination of radionuclidic purity and half-life of the beta emitter 177gLu used for metabolic radioimmunotherapy is presented. High-resolution gamma spectrometry, as well as deconvolution of beta decay curve measured by spectrometry with liquid scintillation counting, have been adopted for quality control of different samples available on the market. A simple method was developed to distinguish between the different methods available for production of 177gLu: i.e. either direct (n,γ) reactions of enriched 176Lu or indirect (n,γ) activation of enriched 176Yb followed by β- decay. In the first case, the long-lived metastable level 177mLu is present in the radioactive preparation and a low specific activity radionuclide is obtained, in the latter a very high purity and high specific activity 177gLu is produced.

  13. Ion chromatography as highly suitable method for rapid and accurate determination of antibiotic fosfomycin in pharmaceutical wastewater.

    PubMed

    Zeng, Ping; Xie, Xiaolin; Song, Yonghui; Liu, Ruixia; Zhu, Chaowei; Galarneau, Anne; Pic, Jean-Stéphane

    2014-01-01

    A rapid and accurate ion chromatography (IC) method (limit of detection as low as 0.06 mg L(-1)) for fosfomycin concentration determination in pharmaceutical industrial wastewater was developed. This method was compared with the performance of high performance liquid chromatography determination (with a high detection limit of 96.0 mg L(-1)) and ultraviolet spectrometry after reacting with alizarin (difficult to perform in colored solutions). The accuracy of the IC method was established in the linear range of 1.0-15.0 mg L(-1) and a linear correlation was found with a correlation coefficient of 0.9998. The recoveries of fosfomycin from industrial pharmaceutical wastewater at spiking concentrations of 2.0, 5.0 and 8.0 mg L(-1) ranged from 81.91 to 94.74%, with a relative standard deviation (RSD) from 1 to 4%. The recoveries of effluent from a sequencing batch reactor treated fosfomycin with activated sludge at spiking concentrations of 5.0, 8.0, 10.0 mg L(-1) ranging from 98.25 to 99.91%, with a RSD from 1 to 2%. The developed IC procedure provided a rapid, reliable and sensitive method for the determination of fosfomycin concentration in industrial pharmaceutical wastewater and samples containing complex components.

  14. Quantitative FRET (Förster Resonance Energy Transfer) analysis for SENP1 protease kinetics determination.

    PubMed

    Liu, Yan; Liao, Jiayu

    2013-02-21

    self-fluorescence and thus was not accurate. We developed a novel highly sensitive and quantitative FRET-based protease assay for determining the kinetic parameters of pre-SUMO1 maturation by SENP1. An engineered FRET pair CyPet and YPet with significantly improved FRET efficiency and fluorescence quantum yield, were used to generate the CyPet-(pre-SUMO1)-YPet substrate (16). We differentiated and quantified absolute fluorescence signals contributed by the donor and acceptor and FRET at the acceptor and emission wavelengths, respectively. The value of kcat/KM was obtained as (3.2 ± 0.55) x10(7) M(-1)s(-1) of SENP1 toward pre-SUMO1, which is in agreement with general enzymatic kinetic parameters. Therefore, this methodology is valid and can be used as a general approach to characterize other proteases as well.

  15. The quantitative determination of FeS2 phases in coal by means of 57Fe Mössbauer spectroscopy

    USGS Publications Warehouse

    Evans, B.J.; King, Hobart M.; Renton, John J.; Stiller, A.

    1990-01-01

    A knowledge of the concentration of pyrite and marcasite in coals can provide important insight into the genesis of coal deposits. Determinations of the relative amounts of pyrite and marcasite by traditional methods of coal analysis are, however, beset with many difficulties. Using 57Fe Mössbauer spectroscopy and a mild chemical treatment with hydrofluoric acid, a technique has been devised for the quantitative determination of the relative concentrations of pyrite and marcasite in samples of whole coals or their low-temperature ashes. The sample preparation procedure is comparable to less accurate methods. Good qualitative agreement has been obtained between ore microscopic and Mössbauer spectroscopic techniques for a series of extensively investigated whole coal samples.

  16. Under proper control, oxidation of proteins with known chemical structure provides an accurate and absolute method for the determination of their molar concentration.

    PubMed

    Guermant, C; Azarkan, M; Smolders, N; Baeyens-Volant, D; Nijs, M; Paul, C; Brygier, J; Vincentelli, J; Looze, Y

    2000-01-01

    Oxidation at 120 degrees C of inorganic and organic (including amino acids, di- and tripeptides) model compounds by K(2)Cr(2)O(7) in the presence of H(2)SO(4) (mass fraction: 0.572), Ag(2)SO(4) (catalyst), and HgSO(4) results in the quantitative conversion of their C-atoms into CO(2) within 24 h or less. Under these stressed, well-defined conditions, the S-atoms present in cysteine and cystine residues are oxidized into SO(3) while, interestingly, the oxidation states of all the other (including the N-) atoms normally present in a protein do remain quite unchanged. When the chemical structure of a given protein is available, the total number of electrons the protein is able to transfer to K(2)Cr(2)O(7) and thereof, the total number of moles of Cr(3+) ions which the protein is able to generate upon oxidation can be accurately calculated. In such cases, unknown protein molar concentrations can thus be determined through straightforward spectrophotometric measurements of Cr(3+) concentrations. The values of molar absorption coefficients for several well-characterized proteins have been redetermined on this basis and observed to be in excellent agreement with the most precise values reported in the literature, which fully assesses the validity of the method. When applied to highly purified proteins of known chemical structure (more generally of known atomic composition), this method is absolute and accurate (+/-1%). Furthermore, it is well adapted to series measurements since available commercial kits for chemical oxygen demand (COD) measurements can readily be adapted to work under the experimental conditions recommended here for the protein assay. PMID:10610688

  17. A vector boundary matching technique for efficient and accurate determination of photonic bandgaps in photonic bandgap fibers.

    PubMed

    Dong, Liang

    2011-06-20

    A vector boundary matching technique has been proposed and demonstrated for finding photonic bandgaps in photonic bandgap fibers with circular nodes. Much improved accuracy, comparing to earlier works, comes mostly from using more accurate cell boundaries for each mode at the upper and lower edges of the band of modes. It is recognized that the unit cell boundary used for finding each mode at band edges of the 2D cladding lattice is not only dependent on whether it is a mode at upper or lower band edge, but also on the azimuthal mode number and lattice arrangements. Unit cell boundaries for these modes are determined by mode symmetries which are governed by the azimuthal mode number as well as lattice arrangement due to mostly geometrical constrains. Unit cell boundaries are determined for modes at both upper and lower edges of bands of modes dominated by m = 1 and m = 2 terms in their longitudinal field Fourier-Bessel expansion series, equivalent to LP0s and LP1s modes in the approximate LP mode representations, for hexagonal lattice to illustrate the technique. The novel technique is also implemented in vector form and incorporates a transfer matrix algorithm for the consideration of nodes with arbitrary refractive index profiles. Both are desired new capabilities for further explorations of advanced new designs of photonic bandgap fibers. PMID:21716499

  18. Nonlinear Optical Properties of Fluorescent Dyes Allow for Accurate Determination of Their Molecular Orientations in Phospholipid Membranes.

    PubMed

    Timr, Štěpán; Brabec, Jiří; Bondar, Alexey; Ryba, Tomáš; Železný, Miloš; Lazar, Josef; Jungwirth, Pavel

    2015-07-30

    Several methods based on single- and two-photon fluorescence detected linear dichroism have recently been used to determine the orientational distributions of fluorescent dyes in lipid membranes. However, these determinations relied on simplified descriptions of nonlinear anisotropic properties of the dye molecules, using a transition dipole-moment-like vector instead of an absorptivity tensor. To investigate the validity of the vector approximation, we have now carried out a combination of computer simulations and polarization microscopy experiments on two representative fluorescent dyes (DiI and F2N12S) embedded in aqueous phosphatidylcholine bilayers. Our results indicate that a simplified vector-like treatment of the two-photon transition tensor is applicable for molecular geometries sampled in the membrane at ambient conditions. Furthermore, our results allow evaluation of several distinct polarization microscopy techniques. In combination, our results point to a robust and accurate experimental and computational treatment of orientational distributions of DiI, F2N12S, and related dyes (including Cy3, Cy5, and others), with implications to monitoring physiologically relevant processes in cellular membranes in a novel way.

  19. Quantitative determination based on the differences between spectra-temperature relationships.

    PubMed

    Li, Zhe; Zhou, Mei; Luo, Yongshun; Li, Gang; Lin, Ling

    2016-08-01

    In the Near-infrared (NIR) spectral measurement it is not always possible to keep the experimental conditions constant. The fluctuations in external variables, such as temperature, will result in a nonlinear shift and a broadening of the spectral bands. In this study, the temperature-induced spectral variation coefficient (TSVC) was obtained by using loading space standardization (LSS). The relationship between TSVC and normalized squared temperature was quantitatively analyzed and applied to the quantitative determination of the compositions in mixtures. NIR spectra of peanut-soy-corn oil mixtures measured at seven temperatures were analyzed. It was found that, the relationship between TSVC and normalized squared temperature can be established by using LSS. Furthermore, the quantitative determination of the compositions in a mixture can be achieved by using the difference between the relationships, i.e., the slope of the relationship. The calibration curves between slope and composition volume are found to be reliable with the correlation coefficients (R(2)) as high as 0.9992. Quantitative determination by the calibration curves were also validated. Therefore, the method can be an effective tool for investigating the effect of temperature and quantitatively analysis. PMID:27216655

  20. High Fidelity Non-Gravitational Force Models for Precise and Accurate Orbit Determination of TerraSAR-X

    NASA Astrophysics Data System (ADS)

    Hackel, Stefan; Montenbruck, Oliver; Steigenberger, -Peter; Eineder, Michael; Gisinger, Christoph

    Remote sensing satellites support a broad range of scientific and commercial applications. The two radar imaging satellites TerraSAR-X and TanDEM-X provide spaceborne Synthetic Aperture Radar (SAR) and interferometric SAR data with a very high accuracy. The increasing demand for precise radar products relies on sophisticated validation methods, which require precise and accurate orbit products. Basically, the precise reconstruction of the satellite’s trajectory is based on the Global Positioning System (GPS) measurements from a geodetic-grade dual-frequency receiver onboard the spacecraft. The Reduced Dynamic Orbit Determination (RDOD) approach utilizes models for the gravitational and non-gravitational forces. Following a proper analysis of the orbit quality, systematics in the orbit products have been identified, which reflect deficits in the non-gravitational force models. A detailed satellite macro model is introduced to describe the geometry and the optical surface properties of the satellite. Two major non-gravitational forces are the direct and the indirect Solar Radiation Pressure (SRP). Due to the dusk-dawn orbit configuration of TerraSAR-X, the satellite is almost constantly illuminated by the Sun. Therefore, the direct SRP has an effect on the lateral stability of the determined orbit. The indirect effect of the solar radiation principally contributes to the Earth Radiation Pressure (ERP). The resulting force depends on the sunlight, which is reflected by the illuminated Earth surface in the visible, and the emission of the Earth body in the infrared spectra. Both components of ERP require Earth models to describe the optical properties of the Earth surface. Therefore, the influence of different Earth models on the orbit quality is assessed within the presentation. The presentation highlights the influence of non-gravitational force and satellite macro models on the orbit quality of TerraSAR-X.

  1. Accurate Focal Depth Determination of Oceanic Earthquakes Using Water-column Reverberation and Some Implications for the Shrinking Plate Hypothesis

    NASA Astrophysics Data System (ADS)

    Niu, F.; Huang, J.; Gordon, R. G.

    2015-12-01

    Investigation of oceanic earthquakes can play an important role in constraining the lateral and depth variations of the stress and strain-rate fields in oceanic lithosphere and of the thickness of the seismogenic layer as a function of lithosphere age, thereby providing us with critical insight into thermal and dynamic processes associated with the cooling and evolution of oceanic lithosphere. With the goal of estimating hypocentral depths more accurately, we observe clear water reverberations after the direct P wave on teleseismic records of oceanic earthquakes and develop a technique to estimate earthquake depths by using these reverberations. The Z-H grid search method allows the simultaneous determination of the sea floor depth (H) and earthquake depth (Z) with an uncertainty less than 1 km, which compares favorably with alternative approaches. We apply this method to two closely located earthquakes beneath the eastern Pacific. These earthquakes occur in ≈25 Ma-old lithosphere and were previously estimated to have very similar depths of ≈10-12 km. We find that the two events actually occurred at dissimilar depths of 2.5 km and 16.8 km beneath the seafloor, respectively within the oceanic crust and lithospheric mantle. The shallow and deep events are determined to be a thrust and normal earthquake, respectively, indicating that the stress field within the oceanic lithosphere changes from horizontal compression to horizontal extension as depth increases, which is consistent with the prediction of the lithospheric cooling model. Furthermore, we show that the P-axis of the newly investigated thrust-faulting earthquake is roughly perpendicular to that of the previously studied thrust event, consistent with the predictions of the shrinking-plate hypothesis.

  2. Accurate focal depth determination of oceanic earthquakes using water-column reverberation and some implications for the shrinking plate hypothesis

    NASA Astrophysics Data System (ADS)

    Huang, Jianping; Niu, Fenglin; Gordon, Richard G.; Cui, Chao

    2015-12-01

    Investigation of oceanic earthquakes is useful for constraining the lateral and depth variations of the stress and strain-rate fields in oceanic lithosphere, and the thickness of the seismogenic layer as a function of lithosphere age, thereby providing us with critical insight into thermal and dynamic processes associated with the cooling and evolution of oceanic lithosphere. With the goal of estimating hypocentral depths more accurately, we observe clear water reverberations after the direct P wave on teleseismic records of oceanic earthquakes and develop a technique to estimate earthquake depths by using these reverberations. The Z-H grid search method allows the simultaneous determination of the sea floor depth (H) and earthquake depth (Z) with an uncertainty less than 1 km, which compares favorably with alternative approaches. We apply this method to two closely located earthquakes beneath the eastern Pacific. These earthquakes occurred in ∼25 Ma-old lithosphere and were previously estimated to have similar depths of ∼10-12 km. We find that the two events actually occurred at dissimilar depths of 2.5 km and 16.8 km beneath the seafloor, respectively, within the oceanic crust and lithospheric mantle. The shallow and deep events are determined to be a thrust and normal earthquake, respectively, indicating that the stress field within the oceanic lithosphere changes from horizontal deviatoric compression to horizontal deviatoric tension as depth increases, which is consistent with the prediction of lithospheric cooling models. Furthermore, we show that the P-axis of the newly investigated thrust-faulting earthquake is perpendicular to that of the previously studied thrust event, consistent with the predictions of the shrinking-plate hypothesis.

  3. Development and validation of a novel, simple, and accurate spectrophotometric method for the determination of lead in human serum.

    PubMed

    Shayesteh, Tavakol Heidari; Khajavi, Farzad; Khosroshahi, Abolfazl Ghafuri; Mahjub, Reza

    2016-01-01

    The determination of blood lead levels is the most useful indicator of the determination of the amount of lead that is absorbed by the human body. Various methods, like atomic absorption spectroscopy (AAS), have already been used for the detection of lead in biological fluid, but most of these methods are based on complicated, expensive, and highly instructed instruments. In this study, a simple and accurate spectroscopic method for the determination of lead has been developed and applied for the investigation of lead concentration in biological samples. In this study, a silica gel column was used to extract lead and eliminate interfering agents in human serum samples. The column was washed with deionized water. The pH was adjusted to the value of 8.2 using phosphate buffer, and then tartrate and cyanide solutions were added as masking agents. The lead content was extracted into the organic phase containing dithizone as a complexion reagent and the dithizone-Pb(II) complex was formed and approved by visible spectrophotometry at 538 nm. The recovery was found to be 84.6 %. In order to validate the method, a calibration curve involving the use of various concentration levels was calculated and proven to be linear in the range of 0.01-1.5 μg/ml, with an R (2) regression coefficient of 0.9968 by statistical analysis of linear model validation. The largest error % values were found to be -5.80 and +11.6 % for intra-day and inter-day measurements, respectively. The largest RSD % values were calculated to be 6.54 and 12.32 % for intra-day and inter-day measurements, respectively. Further, the limit of detection (LOD) was calculated to be 0.002 μg/ml. The developed method was applied to determine the lead content in the human serum of voluntary miners, and it has been proven that there is no statistically significant difference between the data provided from this novel method and the data obtained from previously studied AAS. PMID:26631397

  4. Development and validation of a novel, simple, and accurate spectrophotometric method for the determination of lead in human serum.

    PubMed

    Shayesteh, Tavakol Heidari; Khajavi, Farzad; Khosroshahi, Abolfazl Ghafuri; Mahjub, Reza

    2016-01-01

    The determination of blood lead levels is the most useful indicator of the determination of the amount of lead that is absorbed by the human body. Various methods, like atomic absorption spectroscopy (AAS), have already been used for the detection of lead in biological fluid, but most of these methods are based on complicated, expensive, and highly instructed instruments. In this study, a simple and accurate spectroscopic method for the determination of lead has been developed and applied for the investigation of lead concentration in biological samples. In this study, a silica gel column was used to extract lead and eliminate interfering agents in human serum samples. The column was washed with deionized water. The pH was adjusted to the value of 8.2 using phosphate buffer, and then tartrate and cyanide solutions were added as masking agents. The lead content was extracted into the organic phase containing dithizone as a complexion reagent and the dithizone-Pb(II) complex was formed and approved by visible spectrophotometry at 538 nm. The recovery was found to be 84.6 %. In order to validate the method, a calibration curve involving the use of various concentration levels was calculated and proven to be linear in the range of 0.01-1.5 μg/ml, with an R (2) regression coefficient of 0.9968 by statistical analysis of linear model validation. The largest error % values were found to be -5.80 and +11.6 % for intra-day and inter-day measurements, respectively. The largest RSD % values were calculated to be 6.54 and 12.32 % for intra-day and inter-day measurements, respectively. Further, the limit of detection (LOD) was calculated to be 0.002 μg/ml. The developed method was applied to determine the lead content in the human serum of voluntary miners, and it has been proven that there is no statistically significant difference between the data provided from this novel method and the data obtained from previously studied AAS.

  5. Caregiver's Country of Birth Is a Significant Determinant of Accurate Perception of Preschool-Age Children's Weight

    ERIC Educational Resources Information Center

    Natale, Ruby; Uhlhorn, Susan B.; Lopez-Mitnik, Gabriela; Camejo, Stephanie; Englebert, Nicole; Delamater, Alan M.; Messiah, Sarah E.

    2016-01-01

    Background: One in four preschool-age children in the United States are currently overweight or obese. Previous studies have shown that caregivers of this age group often have difficulty accurately recognizing their child's weight status. The purpose of this study was to examine factors associated with accurate/inaccurate perception of child body…

  6. On the feasibility of quantitative ultrasonic determination of fracture toughness: A literature review

    NASA Technical Reports Server (NTRS)

    Fu, L. S.

    1980-01-01

    The three main topics covered are: (1) fracture toughness and microstructure, (2) quantitative ultrasonic and microstructure; and (3) scattering and related mathematical methods. Literature in these areas is reviewed to give insight to the search of a theoretical foundation for quantitative ultrasonic measurement of fracture toughness. The literature review shows that fracture toughness is inherently related to the microstructure and in particular, it depends upon the spacing of inclusions or second particles and the aspect ratio of second phase particles. There are indications that ultrasonic velocity attenuation measurements can be used to determine fracture toughness. The leads to a review of the mathematical models available in solving boundary value problems related to microstructural factors that govern facture toughness and wave motion. A framework towards the theoretical study for the quantitative determination of fracture toughness is described and suggestions for future research are proposed.

  7. Accurate determination of selected pesticides in soya beans by liquid chromatography coupled to isotope dilution mass spectrometry.

    PubMed

    Huertas Pérez, J F; Sejerøe-Olsen, B; Fernández Alba, A R; Schimmel, H; Dabrio, M

    2015-05-01

    A sensitive, accurate and simple liquid chromatography coupled with mass spectrometry method for the determination of 10 selected pesticides in soya beans has been developed and validated. The method is intended for use during the characterization of selected pesticides in a reference material. In this process, high accuracy and appropriate uncertainty levels associated to the analytical measurements are of utmost importance. The analytical procedure is based on sample extraction by the use of a modified QuEChERS (quick, easy, cheap, effective, rugged, safe) extraction and subsequent clean-up of the extract with C18, PSA and Florisil. Analytes were separated on a C18 column using gradient elution with water-methanol/2.5 mM ammonium acetate mobile phase, and finally identified and quantified by triple quadrupole mass spectrometry in the multiple reaction monitoring mode (MRM). Reliable and accurate quantification of the analytes was achieved by means of stable isotope-labelled analogues employed as internal standards (IS) and calibration with pure substance solutions containing both, the isotopically labelled and native compounds. Exceptions were made for thiodicarb and malaoxon where the isotopically labelled congeners were not commercially available at the time of analysis. For the quantification of those compounds methomyl-(13)C2(15)N and malathion-D10 were used respectively. The method was validated according to the general principles covered by DG SANCO guidelines. However, validation criteria were set more stringently. Mean recoveries were in the range of 86-103% with RSDs lower than 8.1%. Repeatability and intermediate precision were in the range of 3.9-7.6% and 1.9-8.7% respectively. LODs were theoretically estimated and experimentally confirmed to be in the range 0.001-0.005 mg kg(-1) in the matrix, while LOQs established as the lowest spiking mass fractionation level were in the range 0.01-0.05 mg kg(-1). The method reliably identifies and quantifies the

  8. Determination of contents of four alkaloids in Pericarpium arecae by quantitative analysis of multi-components by single-marker.

    PubMed

    Yu, Hongli; Tang, Lijuan; Wu, Hao; Wang, Kuilong; Cai, Baochang; Zhang, Xingde; Zhang, Chengchao; Yin, Yunzhi

    2016-07-01

    By using a typical component in traditional Chinese medicine Pericarpium Arecae (PA), quantitative analysis of multi-components by single-marker (QAMS) was performed to determine the contents of four alkaloids. With a column packed with strong cation exchange bonded silica particles, the alkaloids were well separated, showing linear relationships within certain ranges. The limit of detection, limit of quantitation, precision, stability, repeatability and recovery all met requirements. By employing arecoline as internal standard, relative correction factors for arecaidine, guvacine and guvacoline at five concentrations were detected with three HPLC systems and three HPLC columns. The peaks of arecaidine, guvacine and guvacoline were positioned, during which the columns with the same packing materials from different manufacturers significantly affected relative retention values and retention time differences of the alkaloids. However, the columns, from different batches, managed to give relative retention values satisfying the requirements of HPLC peak positioning. The Thermo Fisher Scientific column packed with strong cation exchange bonded silica particles was finally selected by considering resolution and peak time. Compared with the external standard method, QAMS detected the alkaloid contents in 12 PA samples more accurately and reliably. The results provide valuable evidence for content determination and quality control of alkaloids in PA.

  9. A reference-gene-based quantitative PCR method as a tool to determine Fusarium resistance in wheat.

    PubMed

    Brunner, Kurt; Kovalsky Paris, Maria P; Paolino, Guadalupe; Bürstmayr, Hermann; Lemmens, Marc; Berthiller, Franz; Schuhmacher, Rainer; Krska, Rudolf; Mach, Robert L

    2009-11-01

    In recent years, plant breeders made great progress in breeding Fusarium-tolerant wheat lines. However, total resistance to this genus of plant pathogenic fungi has not yet been achieved as the resistance genes are located on several distinct genetic regions. Visual scoring of disease symptoms in combination with the analysis of mycotoxins is commonly applied to assess the tolerance of new lines. Both approaches are indirect methods and do not mandatorily determine the accumulated fungal biomass. Quantitative PCR is a useful tool to assess fungal biomass based on the abundance of organism-specific DNA. The aim of this study was the development of a quantitative PCR assay for trichothecene-producing Fusarium species and to adapt this method for resistance assessment of wheat lines artificially infected with Fusarium graminearum and Fusarium culmorum. Several DNA-extraction methods for wheat samples were evaluated and optimized for downstream real-time PCR analysis and furthermore, a new reference-gene-based approach for more accurate quantification of Fusarium biomass in cereals is presented. The co-determination of a plant gene was used to compensate for unequal DNA-extraction efficiencies.

  10. Determination of contents of four alkaloids in Pericarpium arecae by quantitative analysis of multi-components by single-marker.

    PubMed

    Yu, Hongli; Tang, Lijuan; Wu, Hao; Wang, Kuilong; Cai, Baochang; Zhang, Xingde; Zhang, Chengchao; Yin, Yunzhi

    2016-07-01

    By using a typical component in traditional Chinese medicine Pericarpium Arecae (PA), quantitative analysis of multi-components by single-marker (QAMS) was performed to determine the contents of four alkaloids. With a column packed with strong cation exchange bonded silica particles, the alkaloids were well separated, showing linear relationships within certain ranges. The limit of detection, limit of quantitation, precision, stability, repeatability and recovery all met requirements. By employing arecoline as internal standard, relative correction factors for arecaidine, guvacine and guvacoline at five concentrations were detected with three HPLC systems and three HPLC columns. The peaks of arecaidine, guvacine and guvacoline were positioned, during which the columns with the same packing materials from different manufacturers significantly affected relative retention values and retention time differences of the alkaloids. However, the columns, from different batches, managed to give relative retention values satisfying the requirements of HPLC peak positioning. The Thermo Fisher Scientific column packed with strong cation exchange bonded silica particles was finally selected by considering resolution and peak time. Compared with the external standard method, QAMS detected the alkaloid contents in 12 PA samples more accurately and reliably. The results provide valuable evidence for content determination and quality control of alkaloids in PA. PMID:27393441

  11. Quantitatively determined self-incompatibility : 3. Genetical variability in Borago officinalis.

    PubMed

    Leach, C R; Mayo, O

    1991-05-01

    It is shown by simulation that a hypothetical multilocus, quantitatively determined self-incompatibility system, whether gametophytic or sporophytic, should maintain variability in small populations at a higher level than would panmixia. Studies of more than 20 isozyme loci show that borage has almost no variability.

  12. Antimicrobial screening and quantitative determination of benzoic acid derivative of Gomphrena celosioides by TLC-densitometry.

    PubMed

    de Moura, Rute Mendonça Xavier; Pereira, Paulo Sérgio; Januário, Ana Helena; França, Suzelei de Castro; Dias, Diones Aparecida

    2004-11-01

    The antimicrobial activity of ethanolic extract and pure compounds of Gomphrena celosioides have been screened by Kirby-Bauer method. Quantitative determination of 4-hydroxy-3-methoxy-benzoic acid in stems, leaves, flowers and roots was established by TLC-densitometry. Results showed significant activity against Staphylococcus aureus and Salmonella typhi. There were no significant differences in the determined benzoic acid derivative. PMID:15516759

  13. Rapid Quantitative Determination of Squalene in Shark Liver Oils by Raman and IR Spectroscopy.

    PubMed

    Hall, David W; Marshall, Susan N; Gordon, Keith C; Killeen, Daniel P

    2016-01-01

    Squalene is sourced predominantly from shark liver oils and to a lesser extent from plants such as olives. It is used for the production of surfactants, dyes, sunscreen, and cosmetics. The economic value of shark liver oil is directly related to the squalene content, which in turn is highly variable and species-dependent. Presented here is a validated gas chromatography-mass spectrometry analysis method for the quantitation of squalene in shark liver oils, with an accuracy of 99.0 %, precision of 0.23 % (standard deviation), and linearity of >0.999. The method has been used to measure the squalene concentration of 16 commercial shark liver oils. These reference squalene concentrations were related to infrared (IR) and Raman spectra of the same oils using partial least squares regression. The resultant models were suitable for the rapid quantitation of squalene in shark liver oils, with cross-validation r (2) values of >0.98 and root mean square errors of validation of ≤4.3 % w/w. Independent test set validation of these models found mean absolute deviations of the 4.9 and 1.0 % w/w for the IR and Raman models, respectively. Both techniques were more accurate than results obtained by an industrial refractive index analysis method, which is used for rapid, cheap quantitation of squalene in shark liver oils. In particular, the Raman partial least squares regression was suited to quantitative squalene analysis. The intense and highly characteristic Raman bands of squalene made quantitative analysis possible irrespective of the lipid matrix.

  14. Rapid Quantitative Determination of Squalene in Shark Liver Oils by Raman and IR Spectroscopy.

    PubMed

    Hall, David W; Marshall, Susan N; Gordon, Keith C; Killeen, Daniel P

    2016-01-01

    Squalene is sourced predominantly from shark liver oils and to a lesser extent from plants such as olives. It is used for the production of surfactants, dyes, sunscreen, and cosmetics. The economic value of shark liver oil is directly related to the squalene content, which in turn is highly variable and species-dependent. Presented here is a validated gas chromatography-mass spectrometry analysis method for the quantitation of squalene in shark liver oils, with an accuracy of 99.0 %, precision of 0.23 % (standard deviation), and linearity of >0.999. The method has been used to measure the squalene concentration of 16 commercial shark liver oils. These reference squalene concentrations were related to infrared (IR) and Raman spectra of the same oils using partial least squares regression. The resultant models were suitable for the rapid quantitation of squalene in shark liver oils, with cross-validation r (2) values of >0.98 and root mean square errors of validation of ≤4.3 % w/w. Independent test set validation of these models found mean absolute deviations of the 4.9 and 1.0 % w/w for the IR and Raman models, respectively. Both techniques were more accurate than results obtained by an industrial refractive index analysis method, which is used for rapid, cheap quantitation of squalene in shark liver oils. In particular, the Raman partial least squares regression was suited to quantitative squalene analysis. The intense and highly characteristic Raman bands of squalene made quantitative analysis possible irrespective of the lipid matrix. PMID:26620374

  15. Determination of Baylisascaris schroederi infection in wild giant pandas by an accurate and sensitive PCR/CE-SSCP method.

    PubMed

    Zhang, Wenping; Yie, Shangmian; Yue, Bisong; Zhou, Jielong; An, Renxiong; Yang, Jiangdong; Chen, Wangli; Wang, Chengdong; Zhang, Liang; Shen, Fujun; Yang, Guangyou; Hou, Rong; Zhang, Zhihe

    2012-01-01

    It has been recognized that other than habitat loss, degradation and fragmentation, the infection of the roundworm Baylisascaris schroederi (B. schroederi) is one of the major causes of death in wild giant pandas. However, the prevalence and intensity of the parasite infection has been inconsistently reported through a method that uses sedimentation-floatation followed by a microscope examination. This method fails to accurately determine infection because there are many bamboo residues and/or few B. schroederi eggs in the examined fecal samples. In the present study, we adopted a method that uses PCR and capillary electrophoresis combined with a single-strand conformation polymorphism analysis (PCR/CE-SSCP) to detect B. schroederi infection in wild giant pandas at a nature reserve, and compared it to the traditional microscope approach. The PCR specifically amplified a single band of 279-bp from both fecal samples and positive controls, which was confirmed by sequence analysis to correspond to the mitochondrial COII gene of B. schroederi. Moreover, it was demonstrated that the amount of genomic DNA was linearly correlated with the peak area of the CE-SSCP analysis. Thus, our adopted method can reliably detect the infectious prevalence and intensity of B. schroederi in wild giant pandas. The prevalence of B. schroederi was found to be 54% in the 91 fecal samples examined, and 48% in the fecal samples of 31 identified individual giant pandas. Infectious intensities of the 91 fecal samples were detected to range from 2.8 to 959.2 units/gram, and from 4.8 to 959.2 units/gram in the fecal samples of the 31 identified giant pandas. For comparison, by using the traditional microscope method, the prevalence of B. schroederi was found to be only 33% in the 91 fecal samples, 32% in the fecal samples of the 31 identified giant pandas, and no reliable infectious intensity was observed. PMID:22911871

  16. Determination of Baylisascaris schroederi Infection in Wild Giant Pandas by an Accurate and Sensitive PCR/CE-SSCP Method

    PubMed Central

    Zhang, Wenping; Yie, Shangmian; Yue, Bisong; Zhou, Jielong; An, Renxiong; Yang, Jiangdong; Chen, Wangli; Wang, Chengdong; Zhang, Liang; Shen, Fujun; Yang, Guangyou; Hou, Rong; Zhang, Zhihe

    2012-01-01

    It has been recognized that other than habitat loss, degradation and fragmentation, the infection of the roundworm Baylisascaris schroederi (B. schroederi) is one of the major causes of death in wild giant pandas. However, the prevalence and intensity of the parasite infection has been inconsistently reported through a method that uses sedimentation-floatation followed by a microscope examination. This method fails to accurately determine infection because there are many bamboo residues and/or few B. schroederi eggs in the examined fecal samples. In the present study, we adopted a method that uses PCR and capillary electrophoresis combined with a single-strand conformation polymorphism analysis (PCR/CE-SSCP) to detect B. schroederi infection in wild giant pandas at a nature reserve, and compared it to the traditional microscope approach. The PCR specifically amplified a single band of 279-bp from both fecal samples and positive controls, which was confirmed by sequence analysis to correspond to the mitochondrial COII gene of B. schroederi. Moreover, it was demonstrated that the amount of genomic DNA was linearly correlated with the peak area of the CE-SSCP analysis. Thus, our adopted method can reliably detect the infectious prevalence and intensity of B. schroederi in wild giant pandas. The prevalence of B. schroederi was found to be 54% in the 91 fecal samples examined, and 48% in the fecal samples of 31 identified individual giant pandas. Infectious intensities of the 91 fecal samples were detected to range from 2.8 to 959.2 units/gram, and from 4.8 to 959.2 units/gram in the fecal samples of the 31 identified giant pandas. For comparison, by using the traditional microscope method, the prevalence of B. schroederi was found to be only 33% in the 91 fecal samples, 32% in the fecal samples of the 31 identified giant pandas, and no reliable infectious intensity was observed. PMID:22911871

  17. Determination of Baylisascaris schroederi infection in wild giant pandas by an accurate and sensitive PCR/CE-SSCP method.

    PubMed

    Zhang, Wenping; Yie, Shangmian; Yue, Bisong; Zhou, Jielong; An, Renxiong; Yang, Jiangdong; Chen, Wangli; Wang, Chengdong; Zhang, Liang; Shen, Fujun; Yang, Guangyou; Hou, Rong; Zhang, Zhihe

    2012-01-01

    It has been recognized that other than habitat loss, degradation and fragmentation, the infection of the roundworm Baylisascaris schroederi (B. schroederi) is one of the major causes of death in wild giant pandas. However, the prevalence and intensity of the parasite infection has been inconsistently reported through a method that uses sedimentation-floatation followed by a microscope examination. This method fails to accurately determine infection because there are many bamboo residues and/or few B. schroederi eggs in the examined fecal samples. In the present study, we adopted a method that uses PCR and capillary electrophoresis combined with a single-strand conformation polymorphism analysis (PCR/CE-SSCP) to detect B. schroederi infection in wild giant pandas at a nature reserve, and compared it to the traditional microscope approach. The PCR specifically amplified a single band of 279-bp from both fecal samples and positive controls, which was confirmed by sequence analysis to correspond to the mitochondrial COII gene of B. schroederi. Moreover, it was demonstrated that the amount of genomic DNA was linearly correlated with the peak area of the CE-SSCP analysis. Thus, our adopted method can reliably detect the infectious prevalence and intensity of B. schroederi in wild giant pandas. The prevalence of B. schroederi was found to be 54% in the 91 fecal samples examined, and 48% in the fecal samples of 31 identified individual giant pandas. Infectious intensities of the 91 fecal samples were detected to range from 2.8 to 959.2 units/gram, and from 4.8 to 959.2 units/gram in the fecal samples of the 31 identified giant pandas. For comparison, by using the traditional microscope method, the prevalence of B. schroederi was found to be only 33% in the 91 fecal samples, 32% in the fecal samples of the 31 identified giant pandas, and no reliable infectious intensity was observed.

  18. A gel-free MS-based quantitative proteomic approach accurately measures cytochrome P450 protein concentrations in human liver microsomes.

    PubMed

    Wang, Michael Zhuo; Wu, Judy Qiju; Dennison, Jennifer B; Bridges, Arlene S; Hall, Stephen D; Kornbluth, Sally; Tidwell, Richard R; Smith, Philip C; Voyksner, Robert D; Paine, Mary F; Hall, James Edwin

    2008-10-01

    The human cytochrome P450 (P450) superfamily consists of membrane-bound proteins that metabolize a myriad of xenobiotics and endogenous compounds. Quantification of P450 expression in various tissues under normal and induced conditions has an important role in drug safety and efficacy. Conventional immunoquantification methods have poor dynamic range, low throughput, and a limited number of specific antibodies. Recent advances in MS-based quantitative proteomics enable absolute protein quantification in a complex biological mixture. We have developed a gel-free MS-based protein quantification strategy to quantify CYP3A enzymes in human liver microsomes (HLM). Recombinant protein-derived proteotypic peptides and synthetic stable isotope-labeled proteotypic peptides were used as calibration standards and internal standards, respectively. The lower limit of quantification was approximately 20 fmol P450. In two separate panels of HLM examined (n = 11 and n = 22), CYP3A, CYP3A4 and CYP3A5 concentrations were determined reproducibly (CV or=0.87) and marker activities (r(2)>or=0.88), including testosterone 6beta-hydroxylation (CYP3A), midazolam 1'-hydroxylation (CYP3A), itraconazole 6-hydroxylation (CYP3A4) and CYP3A5-mediated vincristine M1 formation (CYP3A5). Taken together, our MS-based method provides a specific, sensitive and reliable means of P450 protein quantification and should facilitate P450 characterization during drug development, especially when specific substrates and/or antibodies are unavailable.

  19. Quantitative Doppler-Echocardiographic Determination of Regurgitant Volume in Patients with Aortic Insufficiency

    PubMed Central

    Schoenhagen, Paul; Drude, Ludwig; Klein, Hermann H; Garcia, Mario J

    2008-01-01

    Background: The severity of aortic regurgitation (AR) can be determined by invasive or echocardiographic methods. We systematically compared quantitative invasive and echocardiographic data with semiquantitative invasive grades in a prospective series of patients. Methods: Using Doppler-echocardiography we determined the cardiac output over the aortic, pulmonary and mitral valve in 27 patients (20 with, 7 without AR). Aortic regurgitant volume was calculated as the difference between the cardiac output over aortic and pulmonary valve/ mitral valve. During angiography the severity of AR was assessed semiquantitatively by aortography and the regurgitant volume was calculated invasively as the difference between the left- and right ventricular cardiac output. Results: The echocardiographically and invasively determined regurgitant blood volume correlated closely (R≈0.8). The regurgitant volume increased with higher angiographic grade but there was significant overlap between adjoining qualitative grades. Conclusion: In patients with AR, quantitative echocardiographic and angiographic measurements of the regurgitant volume correlate closely. PMID:19590613

  20. DeconMSn: A Software Tool for accurate parent ion monoisotopic mass determination for tandem mass spectra

    SciTech Connect

    Mayampurath, Anoop M.; Jaitly, Navdeep; Purvine, Samuel O.; Monroe, Matthew E.; Auberry, Kenneth J.; Adkins, Joshua N.; Smith, Richard D.

    2008-04-01

    We present a new software tool for tandem MS analyses that: • accurately calculates the monoisotopic mass and charge of high–resolution parent ions • accurately operates regardless of the mass selected for fragmentation • performs independent of instrument settings • enables optimal selection of search mass tolerance for high mass accuracy experiments • is open source and thus can be tailored to individual needs • incorporates a SVM-based charge detection algorithm for analyzing low resolution tandem MS spectra • creates multiple output data formats (.dta, .MGF) • handles .RAW files and .mzXML formats • compatible with SEQUEST, MASCOT, X!Tandem

  1. Fast and accurate determination of K, Ca, and Mg in human serum by sector field ICP-MS.

    PubMed

    Yu, Lee L; Davis, W Clay; Nuevo Ordonez, Yoana; Long, Stephen E

    2013-11-01

    Electrolytes in serum are important biomarkers for skeletal and cellular health. The levels of electrolytes are monitored by measuring the Ca, Mg, K, and Na in blood serum. Many reference methods have been developed for the determination of Ca, Mg, and K in clinical measurements; however, isotope dilution thermal ionization mass spectrometry (ID-TIMS) has traditionally been the primary reference method serving as an anchor for traceability and accuracy to these secondary reference methods. The sample matrix must be separated before ID-TIMS measurements, which is a slow and tedious process that hindered the adoption of the technique in routine clinical measurements. We have developed a fast and accurate method for the determination of Ca, Mg, and K in serum by taking advantage of the higher mass resolution capability of the modern sector field inductively coupled plasma mass spectrometry (SF-ICP-MS). Each serum sample was spiked with a mixture containing enriched (44)Ca, (26)Mg, and (41)K, and the (42)Ca(+):(44)Ca(+), (24)Mg(+):(26)Mg(+), and (39)K(+):(41)K(+) ratios were measured. The Ca and Mg ratios were measured in medium resolution mode (m/Δm ≈ 4 500), and the K ratio in high resolution mode (m/Δm ≈ 10 000). Residual (40)Ar(1)H(+) interference was still observed but the deleterious effects of the interference were minimized by measuring the sample at K > 100 ng g(-1). The interferences of Sr(++) at the two Ca isotopes were less than 0.25 % of the analyte signal, and they were corrected with the (88)Sr(+) intensity by using the Sr(++):Sr(+) ratio. The sample preparation involved only simple dilutions, and the measurement using this sample preparation approach is known as dilution-and-shoot (DNS). The DNS approach was validated with samples prepared via the traditional acid digestion approach followed by ID-SF-ICP-MS measurement. DNS and digested samples of SRM 956c were measured with ID-SF-ICP-MS for quality assurance, and the results (mean

  2. Quantitative determination of van Gogh's painting grounds using SEM/EDX.

    PubMed

    Haswell, Ralph; Carlyle, Leslie; Mensch, Kees T J

    2011-10-01

    We have investigated the potential of utilizing analytical electron microscopy to quantitatively examine the grounds used by van Gogh and, in particular, the absolute amount of extender employed. To determine the accuracy that can be achieved, a series of oil paint reconstructions were used as standards. The proportion of extender was measured using scanning electron microscopy and energy dispersive X-ray spectroscopy, and a relative error of 10% or better was achieved. The same method was then used to determine the ground composition of real samples from van Gogh paintings. The results obtained in this work are part of a more quantitative method of comparing and classifying paint cross sections, which will supplement the more traditional qualitative approach. The information obtained from this study is being used to add to our knowledge of the methods and materials used by van Gogh, which is helping in the reconstruction of van Gogh's oeuvre and attribution. PMID:21144113

  3. Procedure for quantitative determination of effectiveness of photoinduced destruction of malignant tumors

    NASA Astrophysics Data System (ADS)

    Bizyuk, S. A.; Istomin, Yu. P.; Dzhagarov, B. M.

    2006-07-01

    We have developed a procedure for analysis of the functional status of blood vessels in tumor tissues using computer-assisted color scanning of tumor slices and also for a quantitative assessment of the effectiveness of photoinduced destruction of tumor tissues in animal experiments. Its major advantage is direct determination of the size of the tumor necrosis zone. The procedure has been tested in an experiment on three strains of malignant tumors with different morphologies.

  4. Quantitative determination of alpha-tocopherol in Arbutus unedo by TLC-densitometry and colorimetry.

    PubMed

    Kivçak, B; Mert, T

    2001-08-01

    A quantitative determination of alpha-tocopherol in Arbutus unedo leaves was established by TLC-densitometry and colorimetry. Data obtained by TLC-densitometry were compared with those obtained by colorimetry. Also, the alpha-tocopherol content in leaves collected at different times of the year was studied comparatively. The highest amount of alpha-tocopherol was found in the March collection. PMID:11543963

  5. Quantitative determination of oil content in small quantity of oilseed rape by ultrasound-assisted extraction combined with gas chromatography.

    PubMed

    Wei, Fang; Gao, Gui-Zhen; Wang, Xin-Fa; Dong, Xu-Yan; Li, Ping-Ping; Hua, Wei; Wang, Xu; Wu, Xiao-Ming; Chen, Hong

    2008-09-01

    Accurately quantitative determination of oil content in oilseed rape plays an important role in varieties breeding for improving oil content in seeds. However, large quantity of oilseeds were needed in order to obtain accuracy and precision results by using standard Soxhlet extraction method, which may be a handicap in analysis of small, rare and precious samples in plant breeding. In the present work, ultrasound-assisted extraction was evaluated as a simpler and more effective alternative to conventional extraction method for the isolation of oil from small quantity of oilseed rape (<20 mg). The oil of oilseed rape samples was extracted by ultrasound-assisted method, and then the fatty acids and total oil content of the seeds were qualitatively and quantitatively determined by gas chromatography (GC). Extraction efficiency of total oil obtained by ultrasound-assisted extraction through an orthogonal experiment (L(9) (3(4))) were investigated to get the best extraction conditions. Statistical analysis showed that the variable with the largest effect was the ultrasound-assisted extraction time which was followed by the ultrasound-assisted extraction power, and the liquid:solid ratio. A liquid:solid ratio of 1:4 (L:g), an ultrasound-assisted extraction time of 60 min and an ultrasound-assisted extraction power of 500 W were found to be optimal for oil extraction from oilseed rape. By comparing with the conventional method, it was found that the ultrasound-assisted extraction of oil from oilseed rape was about five times faster than the traditional extraction method. By the use of ultrasound-assisted extraction combined with GC analysis, the fatty acids and total oil content in small quantity of seeds (<20 mg) were successfully qualitatively determined and the results are in agreement with that obtained by traditional standard method.

  6. Quantitative radiochemical method for determination of major sources of natural radioactivity in ores and minerals

    USGS Publications Warehouse

    Rosholt, J.N.

    1954-01-01

    When an ore sample contains radioactivity other than that attributable to the uranium series in equilibrium, a quantitative analysis of the other emitters must be made in order to determine the source of this activity. Thorium-232, radon-222, and lead-210 have been determined by isolation and subsequent activity analysis of some of their short-lived daughter products. The sulfides of bismuth and polonium are precipitated out of solutions of thorium or uranium ores, and the ??-particle activity of polonium-214, polonium-212, and polonium-210 is determined by scintillation-counting techniques. Polonium-214 activity is used to determine radon-222, polonium-212 activity for thorium-232, and polonium-210 for lead-210. The development of these methods of radiochemical analysis will facilitate the rapid determination of some of the major sources of natural radioactivity.

  7. Quantitative Förster resonance energy transfer analysis for kinetic determinations of SUMO-specific protease.

    PubMed

    Liu, Yan; Song, Yang; Madahar, Vipul; Liao, Jiayu

    2012-03-01

    Förster resonance energy transfer (FRET) technology has been widely used in biological and biomedical research, and it is a very powerful tool for elucidating protein interactions in either dynamic or steady state. SUMOylation (the process of SUMO [small ubiquitin-like modifier] conjugation to substrates) is an important posttranslational protein modification with critical roles in multiple biological processes. Conjugating SUMO to substrates requires an enzymatic cascade. Sentrin/SUMO-specific proteases (SENPs) act as an endopeptidase to process the pre-SUMO or as an isopeptidase to deconjugate SUMO from its substrate. To fully understand the roles of SENPs in the SUMOylation cycle, it is critical to understand their kinetics. Here, we report a novel development of a quantitative FRET-based protease assay for SENP1 kinetic parameter determination. The assay is based on the quantitative analysis of the FRET signal from the total fluorescent signal at acceptor emission wavelength, which consists of three components: donor (CyPet-SUMO1) emission, acceptor (YPet) emission, and FRET signal during the digestion process. Subsequently, we developed novel theoretical and experimental procedures to determine the kinetic parameters, k(cat), K(M), and catalytic efficiency (k(cat)/K(M)) of catalytic domain SENP1 toward pre-SUMO1. Importantly, the general principles of this quantitative FRET-based protease kinetic determination can be applied to other proteases.

  8. [Diagnostic value of the quantitative HBsAg determination in hepatitis B infections].

    PubMed

    van Helden, J; Weiskirchen, R

    2016-04-01

    Introduction of systematic hepatitis B vaccination has lead to a strong decrease of new infections, but there are still a high numbers of chronically infected persons suffering on long-term complications. Using quantitative assays for the determination of HbsAg (qHBsAg) has improved our understanding of chronic hepatitis B (CHB). The concentrations of HBsAg are strongly varying through the different stages of infection. The quantitative determination of HBsAg does not only yield in additional information to the infection activity, but also provides data for an improved follow up independent from the virus load. As to the prediction of disease progression, low-viremic carriers with high HbsAg levels have been shown to be at higher risk of HBeAg negative hepatitis, cirrhosis and hepatocellular carcinoma (HCC). Although, quantitative HBsAg determination has been widely used in CHB patients receiving pegylated interferon therapy, the HbsAg decline is slow compared to HBV-DNA levels during nucleos(t)ide analogue (NUC) therapy. However a rapid HbsAg decline during NUC therapy may identify patients who will finally clear HbsAg. A 6- to 12-monthly assessment of HbsAg level could be considered during NUC therapy. Taking these lines of evidence together, qHBsAg can complement HBV-DNA levels to optimize the management of CHB patients.

  9. Species Determination and Quantitation in Mixtures Using MRM Mass Spectrometry of Peptides Applied to Meat Authentication.

    PubMed

    Gunning, Yvonne; Watson, Andrew D; Rigby, Neil M; Philo, Mark; Peazer, Joshua K; Kemsley, E Kate

    2016-09-20

    We describe a simple protocol for identifying and quantifying the two components in binary mixtures of species possessing one or more similar proteins. Central to the method is the identification of 'corresponding proteins' in the species of interest, in other words proteins that are nominally the same but possess species-specific sequence differences. When subject to proteolysis, corresponding proteins will give rise to some peptides which are likewise similar but with species-specific variants. These are 'corresponding peptides'. Species-specific peptides can be used as markers for species determination, while pairs of corresponding peptides permit relative quantitation of two species in a mixture. The peptides are detected using multiple reaction monitoring (MRM) mass spectrometry, a highly specific technique that enables peptide-based species determination even in complex systems. In addition, the ratio of MRM peak areas deriving from corresponding peptides supports relative quantitation. Since corresponding proteins and peptides will, in the main, behave similarly in both processing and in experimental extraction and sample preparation, the relative quantitation should remain comparatively robust. In addition, this approach does not need the standards and calibrations required by absolute quantitation methods. The protocol is described in the context of red meats, which have convenient corresponding proteins in the form of their respective myoglobins. This application is relevant to food fraud detection: the method can detect 1% weight for weight of horse meat in beef. The corresponding protein, corresponding peptide (CPCP) relative quantitation using MRM peak area ratios gives good estimates of the weight for weight composition of a horse plus beef mixture.

  10. Species Determination and Quantitation in Mixtures Using MRM Mass Spectrometry of Peptides Applied to Meat Authentication.

    PubMed

    Gunning, Yvonne; Watson, Andrew D; Rigby, Neil M; Philo, Mark; Peazer, Joshua K; Kemsley, E Kate

    2016-01-01

    We describe a simple protocol for identifying and quantifying the two components in binary mixtures of species possessing one or more similar proteins. Central to the method is the identification of 'corresponding proteins' in the species of interest, in other words proteins that are nominally the same but possess species-specific sequence differences. When subject to proteolysis, corresponding proteins will give rise to some peptides which are likewise similar but with species-specific variants. These are 'corresponding peptides'. Species-specific peptides can be used as markers for species determination, while pairs of corresponding peptides permit relative quantitation of two species in a mixture. The peptides are detected using multiple reaction monitoring (MRM) mass spectrometry, a highly specific technique that enables peptide-based species determination even in complex systems. In addition, the ratio of MRM peak areas deriving from corresponding peptides supports relative quantitation. Since corresponding proteins and peptides will, in the main, behave similarly in both processing and in experimental extraction and sample preparation, the relative quantitation should remain comparatively robust. In addition, this approach does not need the standards and calibrations required by absolute quantitation methods. The protocol is described in the context of red meats, which have convenient corresponding proteins in the form of their respective myoglobins. This application is relevant to food fraud detection: the method can detect 1% weight for weight of horse meat in beef. The corresponding protein, corresponding peptide (CPCP) relative quantitation using MRM peak area ratios gives good estimates of the weight for weight composition of a horse plus beef mixture. PMID:27685654

  11. Quantitative Determination of Photosynthetic Pigments in Green Beans Using Thin-Layer Chromatography and a Flatbed Scanner as Densitometer

    ERIC Educational Resources Information Center

    Valverde, Juan; This, Herve; Vignolle, Marc

    2007-01-01

    A simple method for the quantitative determination of photosynthetic pigments extracted from green beans using thin-layer chromatography is proposed. Various extraction methods are compared, and it is shown how a simple flatbed scanner and free software for image processing can give a quantitative determination of pigments. (Contains 5 figures.)

  12. Temperature-dependent turnovers in sex-determination mechanisms: a quantitative model.

    PubMed

    Grossen, Christine; Neuenschwander, Samuel; Perrin, Nicolas

    2011-01-01

    Sex determination is often seen as a dichotomous process: individual sex is assumed to be determined either by genetic (genotypic sex determination, GSD) or by environmental factors (environmental sex determination, ESD), most often temperature (temperature sex determination, TSD). We endorse an alternative view, which sees GSD and TSD as the ends of a continuum. Both effects interact a priori, because temperature can affect gene expression at any step along the sex-determination cascade. We propose to define sex-determination systems at the population- (rather than individual) level, via the proportion of variance in phenotypic sex stemming from genetic versus environmental factors, and we formalize this concept in a quantitative-genetics framework. Sex is seen as a threshold trait underlain by a liability factor, and reaction norms allow modeling interactions between genotypic and temperature effects (seen as the necessary consequences of thermodynamic constraints on the underlying physiological processes). As this formalization shows, temperature changes (due to e.g., climatic changes or range expansions) are expected to provoke turnovers in sex-determination mechanisms, by inducing large-scale sex reversal and thereby sex-ratio selection for alternative sex-determining genes. The frequency of turnovers and prevalence of homomorphic sex chromosomes in cold-blooded vertebrates might thus directly relate to the temperature dependence in sex-determination mechanisms.

  13. Quantitative determination of aflatoxin B1 concentration in acetonitrile by chemometric methods using terahertz spectroscopy.

    PubMed

    Ge, Hongyi; Jiang, Yuying; Lian, Feiyu; Zhang, Yuan; Xia, Shanhong

    2016-10-15

    Aflatoxins contaminate and colonize agricultural products, such as grain, and thereby potentially cause human liver carcinoma. Detection via conventional methods has proven to be time-consuming and complex. In this paper, the terahertz (THz) spectra of aflatoxin B1 in acetonitrile solutions with concentration ranges of 1-50μg/ml and 1-50μg/l are obtained and analyzed for the frequency range of 0.4-1.6THz. Linear and nonlinear regression models are constructed to relate the absorption spectra and the concentrations of 160 samples using the partial least squares (PLS), principal component regression (PCR), support vector machine (SVM), and PCA-SVM methods. Our results indicate that PLS and PCR models are more accurate for the concentration range of 1-50μg/ml, whereas SVM and PCA-SVM are more accurate for the concentration range of 1-50μg/l. Furthermore, ten unknown concentration samples extracted from mildewed maize are analyzed quantitatively using these methods. PMID:27173565

  14. Quantitative Determination of Fluorine Content in Blends of Polylactide (PLA)-Talc Using Near Infrared Spectroscopy.

    PubMed

    Tamburini, Elena; Tagliati, Chiara; Bonato, Tiziano; Costa, Stefania; Scapoli, Chiara; Pedrini, Paola

    2016-08-02

    Near-infrared spectroscopy (NIRS) has been widely used for quantitative and/or qualitative determination of a wide range of matrices. The objective of this study was to develop a NIRS method for the quantitative determination of fluorine content in polylactide (PLA)-talc blends. A blending profile was obtained by mixing different amounts of PLA granules and talc powder. The calibration model was built correlating wet chemical data (alkali digestion method) and NIR spectra. Using FT (Fourier Transform)-NIR technique, a Partial Least Squares (PLS) regression model was set-up, in a concentration interval of 0 ppm of pure PLA to 800 ppm of pure talc. Fluorine content prediction (R²cal = 0.9498; standard error of calibration, SEC = 34.77; standard error of cross-validation, SECV = 46.94) was then externally validated by means of a further 15 independent samples (R²EX.V = 0.8955; root mean standard error of prediction, RMSEP = 61.08). A positive relationship between an inorganic component as fluorine and NIR signal has been evidenced, and used to obtain quantitative analytical information from the spectra.

  15. Quantitative Determination of Fluorine Content in Blends of Polylactide (PLA)-Talc Using Near Infrared Spectroscopy.

    PubMed

    Tamburini, Elena; Tagliati, Chiara; Bonato, Tiziano; Costa, Stefania; Scapoli, Chiara; Pedrini, Paola

    2016-01-01

    Near-infrared spectroscopy (NIRS) has been widely used for quantitative and/or qualitative determination of a wide range of matrices. The objective of this study was to develop a NIRS method for the quantitative determination of fluorine content in polylactide (PLA)-talc blends. A blending profile was obtained by mixing different amounts of PLA granules and talc powder. The calibration model was built correlating wet chemical data (alkali digestion method) and NIR spectra. Using FT (Fourier Transform)-NIR technique, a Partial Least Squares (PLS) regression model was set-up, in a concentration interval of 0 ppm of pure PLA to 800 ppm of pure talc. Fluorine content prediction (R²cal = 0.9498; standard error of calibration, SEC = 34.77; standard error of cross-validation, SECV = 46.94) was then externally validated by means of a further 15 independent samples (R²EX.V = 0.8955; root mean standard error of prediction, RMSEP = 61.08). A positive relationship between an inorganic component as fluorine and NIR signal has been evidenced, and used to obtain quantitative analytical information from the spectra. PMID:27490548

  16. Quantitative determination of bromine and iodine in food samples using ICP-MS.

    PubMed

    Nguyen, Thi Kim Dzung; Ludwig, Rainer

    2014-01-01

    Trace concentrations of bromine and iodine in food samples and certified reference materials (CRMs) were determined by an inductively coupled plasma-mass spectrometry (ICP-MS) technique after low-power microwave digestion and extraction into an aqueous quaternary ammonium hydroxide solution. The recovery after sample preparation was quantitative. The internal standard for the measurement of the analyte on ICP-MS was optimized in this study. The detection limits were 0.19 and 0.68 ng g(-1) for I and Br, respectively, when a 10 ng g(-1) Te solution as an internal standard was used, applying the signal of (125)Te. The high recovery and reproducibility are sufficient for the quantitative analysis of these elements, and the analytical procedure is recommended for the analysis of Br and I in various kinds of bio-samples.

  17. Quantitative determination of local potential values in inhomogeneously doped semiconductors by scanning tunneling microscopy

    NASA Astrophysics Data System (ADS)

    Weidlich, P. H.; Dunin-Borkowski, R. E.; Ebert, Ph.

    2011-08-01

    Local potential changes arising from nanoscale three-dimensional spatial fluctuations in the dopant distribution in Zn-doped GaAs were investigated quantitatively by scanning tunneling microscopy and spectroscopy at (110) cleavage surfaces. Tunneling spectra measured in areas with different local doping concentration show apparent shifts of the valence band edge and apparent changes of the band gap. A quantitative analysis, combined with band bending and tunnel current simulations, demonstrates that these effects arise from tip-induced band bending that modulates the real potential changes. It is illustrated how exact potential changes between locally high and low doped areas can be determined. It is found that the local potential fluctuations in three-dimensionally doped semiconductors are approximately one order of magnitude smaller that those observed in two-dimensionally doped semiconductors.

  18. Determination of the quantitative stereoselectivity fingerprint of lipases during hydrolysis of a prochiral triacylglycerol.

    PubMed

    Mitchell, David Alexander; Rodriguez, Jorge A; Carrière, Frederic; Krieger, Nadia

    2008-06-01

    We propose a method for characterizing quantitatively the stereoselectivity of lipases during hydrolysis of triacylglycerols. Although it is of general applicability, we demonstrate it specifically for sn-1,3-regiospecific lipases. In this case the method generates a "stereoselectivity fingerprint" that consists of ratios of the specificity constants for the various reactions that produce and consume the 1,2-sn- and 2,3-sn-diacylglycerols. We use the method to determine the stereoselectivity fingerprint of several lipases during the hydrolysis of the prochiral substrate triolein. Our method opens up the possibility of correlating quantitative fingerprints with structural information, in the quest to elucidate the mechanisms underlying the stereoselectivity of lipases.

  19. Rapid screening and quantitative determination of bioactive compounds from fruit extracts of Myristica species and their in vitro antiproliferative activity.

    PubMed

    Pandey, Renu; Mahar, Rohit; Hasanain, Mohammad; Shukla, Sanjeev K; Sarkar, Jayanta; Rameshkumar, K B; Kumar, Brijesh

    2016-11-15

    Efficient and sensitive LC-MS/MS methods have been developed for the rapid screening and determination of bioactive compounds in different fruit parts of four Myristica species, viz., Myristica beddomeii, Myristica fragrans, Myristica fatua and Myristica malabarica. Twenty-one compounds were identified and characterized on the basis of their accurate mass and MS/MS fragmentation pattern using HPLC-QTOF-MS/MS and NMR analysis. Quantitative determination of five major bioactive compounds was performed using multiple-reaction monitoring mode with continuous polarity switching by UHPLC-QqQLIT-MS/MS. Moreover, in vitro antiproliferative activity of these Myristica species was evaluated against five human cancer cell lines A549, DLD-1, DU145, FaDu and MCF-7 using SRB assay. Seventeen phytoconstituents were identified and reported for the first time from M. beddomeii and sixteen from M. fatua. Quantification result showed highest total content of five major bioactive compounds in mace of M. fragrans. Evaluation of in vitro antiproliferative activity revealed potent activity in all investigated species except M. fragrans.

  20. Quantitative Determination of L-DOPA in Seeds of Mucuna Pruriens Germplasm by High Performance Thin Layer Chromatography.

    PubMed

    Raina, Archana P; Khatri, Renu

    2011-07-01

    Mucuna pruriens Linn. is an important medicinal plant used for treatment of Parkinson's disease and many others in ancient Indian medical system. L-DOPA extracted from seeds of Mucuna is a constituent of more than 200 indigenous drug formulations and is more effective as drug than the synthetic counterpart. A densitometric high performance thin-layer chromatographic (HPTLC) method was developed for quantification of L-DOPA content present in the seeds extract. The method involves separation of L-DOPA on precoated silica gel 60 GF(254) HPTLC plates using a solvent system of n-butanol-acetic-acid-water (4:1:1, v/v) as the mobile phase. Quantification was done at 280 nm using absorbance reflectance mode. Linearity was found in the concentration range of 100 to 1000 ng/spot with the correlation coefficient value of 0.9980. The method was validated for accuracy, precision and repeatability. Mean recovery was 100.89%. The LOD and LOQ for L-DOPA determination were found to be 3.41 ng/spot and 10.35 ng/spot respectively. The proposed HPTLC method was found to be precise, specific and accurate for quantitative determination of L-DOPA. It can be used for rapid screening of large germplasm collections of Mucuna pruriens for L-DOPA content. The method was used to study variation in fifteen accessions of Mucuna germplasm collected from different geographical regions. PMID:22707835

  1. Rapid screening and quantitative determination of bioactive compounds from fruit extracts of Myristica species and their in vitro antiproliferative activity.

    PubMed

    Pandey, Renu; Mahar, Rohit; Hasanain, Mohammad; Shukla, Sanjeev K; Sarkar, Jayanta; Rameshkumar, K B; Kumar, Brijesh

    2016-11-15

    Efficient and sensitive LC-MS/MS methods have been developed for the rapid screening and determination of bioactive compounds in different fruit parts of four Myristica species, viz., Myristica beddomeii, Myristica fragrans, Myristica fatua and Myristica malabarica. Twenty-one compounds were identified and characterized on the basis of their accurate mass and MS/MS fragmentation pattern using HPLC-QTOF-MS/MS and NMR analysis. Quantitative determination of five major bioactive compounds was performed using multiple-reaction monitoring mode with continuous polarity switching by UHPLC-QqQLIT-MS/MS. Moreover, in vitro antiproliferative activity of these Myristica species was evaluated against five human cancer cell lines A549, DLD-1, DU145, FaDu and MCF-7 using SRB assay. Seventeen phytoconstituents were identified and reported for the first time from M. beddomeii and sixteen from M. fatua. Quantification result showed highest total content of five major bioactive compounds in mace of M. fragrans. Evaluation of in vitro antiproliferative activity revealed potent activity in all investigated species except M. fragrans. PMID:27283658

  2. [Quantitative determination of the infectivity of nuclear polyhedrosis virus DNA on honeycomb moth (Galleria mellonella) larvae].

    PubMed

    Gorbunova, E E; Makarova, N I; Shchelkunova, G A

    1980-01-01

    The possibility of using honeycomb moth larvae for titration of nuclear polyhedrosis virus (NPV) infectious DNA and determinations of transfection effectiveness was studied. Honeycomb moth larvae were shown to be a sensitive system for NPV DNA titration. DEAE-dextran used as a protector increased NPV DNA infectivity 1000-fold, LD50 in this instance being 2 X 10(8) molecules per larva. The method of NPV DNA infectivity determinations by the number of larvae with polyhedreae in the fatty tissue is more sensitive than infectivity determinations by the number of dead larvae and permits titrations of low DNA concentrations. The curve of DNA titration in the presence of DEAE-dextran by the number of larvae with polyhedrae in the fatty tissue allows to quantitate native DNA within the range of 0.01 to 5 micrograms/ml.

  3. 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

    2016-08-01

    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.

  4. Quantitative head ultrasound measurements to determine thresholds for preterm neonates requiring interventional therapies following intraventricular hemorrhage

    NASA Astrophysics Data System (ADS)

    Kishimoto, Jessica; Fenster, Aaron; Salehi, Fateme; Romano, Walter; Lee, David S. C.; de Ribaupierre, Sandrine

    2016-04-01

    Dilation of the cerebral ventricles is a common condition in preterm neonates with intraventricular hemorrhage (IVH). This post hemorrhagic ventricle dilation (PHVD) can lead to lifelong neurological impairment through ischemic injury due to increased intracranial pressure and without treatment, can lead to death. Clinically, 2D ultrasound (US) through the fontanelles ('soft spots') of the patients are serially acquired to monitor the progression of the ventricle dilation. These images are used to determine when interventional therapies such as needle aspiration of the built up cerebrospinal fluid (CSF) ('ventricle tap', VT) might be indicated for a patient; however, quantitative measurements of the growth of the ventricles are often not performed. There is no consensus on when a neonate with PHVD should have an intervention and often interventions are performed after the potential for brain damage is quite high. Previously we have developed and validated a 3D US system to monitor the progression of ventricle volumes (VV) in IVH patients. We will describe the potential utility of quantitative 2D and 3D US to monitor and manage PHVD in neonates. Specifically, we will look to determine image-based measurement thresholds for patients who will require VT in comparison to patients with PHVD who resolve without intervention. Additionally, since many patients who have an initial VT will require subsequent interventions, we look at the potential for US to determine which PHVD patients will require additional VT after the initial one has been performed.

  5. Quantitative determination of G6Pase activity in histochemically defined zones of the liver acinus.

    PubMed

    Teutsch, H F

    1978-12-13

    Qualitative histochemical G6Pase distribution patterns obtained with an improved method (Teutsch, 1978) served as the basis for a zonal microdissection of the liver acinus. G6Pase activity was determined quantitatively in tissue samples of zones 1 and 3 by a microfluorometric method (Burch et al., 1978). Using a correlation system it could be demonstrated that the histochemical distribution pattern obtained with the improved method was in better agreement with quantitatively estimated zonal differences of G6Pase activity, both in fed and starved female rats, than with the Wachstein and Meisel medium (1956). From a total of 50 tissue samples analyzed the following average G6Pase activities were calculated: in fed animals 15.36 +/- 3.48 U/g dry weight in zone 1, and 9.28 +/- 2.15 U/g dry weight in zone 3; in starved female rats 42.50 +/- 8.20 U/g dry weight in zone 1, and 29.25 +/- 5.68 U/g dry weight in zone 3. The qualitative histochemical as well as quantitative zonal differences of G6Pase activities are taken as further support for the hypothesis of metabolic zonation of liver parenchyma.

  6. Quantitative determination of electric field strengths within dynamically operated devices using EBIC analysis in the SEM.

    PubMed

    Pugatschow, Anton; Heiderhoff, Ralf; Balk, Ludwig J

    2008-01-01

    Although electron beam-induced current (EBIC) technique was invented in the seventies, it is still a powerful technique for failure analysis and reliability investigations of modern materials and devices. Time-resolved and stroboscopic microanalyses using sampling Fourier components decomposed by modulated charge carrier excitation are introduced. Quantitative determination of electric field strengths within dynamically operated devices in the scanning electron microscope (SEM) will be demonstrated. This technique allows investigations of diffusion and drift processes and of variations of electric field distributions inside active devices.

  7. Optical fibre spectroscopy sensor for the quantitative determination of industrial textile dyes

    NASA Astrophysics Data System (ADS)

    Cubillas, Ana M.; Conde, Olga M.; Anuarbe, Pedro; Gutierrez, Monica; Martinez, Vicente; Lopez-Higuera, Jose M.

    2009-10-01

    In this paper, an extrinsic optical fibre sensor (OFS) for the quantitative determination of dyes used in the textile industry is presented. The system proposed is based on absorption spectroscopy and multivariate calibration methods to infer the concentration of different textile dyes. The performance of the sensor has been successfully assessed using calibrated dyes, with a very good correlation between the multivariate calibration models and the predicted values. The sensor system here demonstrated could be used to predict the colour of dye mixtures during the dyebath and, therefore, reduce the manufacturing costs.

  8. A rapid, economical, and accurate method to determining the physical risk of storm marine inundations using sedimentary evidence

    NASA Astrophysics Data System (ADS)

    Nott, Jonathan F.

    2015-04-01

    The majority of physical risk assessments from storm surge inundations are derived from synthetic time series generated from short climate records, which can often result in inaccuracies and are time-consuming and expensive to develop. A new method is presented here for the wet tropics region of northeast Australia. It uses lidar-generated topographic cross sections of beach ridge plains, which have been demonstrated to be deposited by marine inundations generated by tropical cyclones. Extreme value theory statistics are applied to data derived from the cross sections to generate return period plots for a given location. The results suggest that previous methods to estimate return periods using synthetic data sets have underestimated the magnitude/frequency relationship by at least an order of magnitude. The new method promises to be a more rapid, economical, and accurate assessment of the physical risk of these events.

  9. Invited review: quantitative trait nucleotide determination in the era of genomic selection.

    PubMed

    Weller, J I; Ron, M

    2011-03-01

    Genome-wide association studies based on tens of thousands of single nucleotide polymorphisms have been completed for several dairy cattle populations. Methods have been proposed to directly incorporate genome scan data into breeding programs, chiefly by selection of young sires based on their genotypes for the genetic markers and pedigree without progeny test. Thus, the rate of genetic gain is increased by reduction of the mean generation interval. The methods developed so far for application of genomic selection do not require identification of the actual quantitative trait nucleotides (QTN) responsible for the observed variation of quantitative trait loci (QTL). To date, 2 QTN affecting milk production traits have been detected in dairy cattle: DGAT1 and ABCG2. This review will attempt to address the following questions based on the current state of bovine genomics and statistics. What are the pros and cons for QTN determination? How can data obtained from high-density, genome-wide scans be used most efficiently for QTN determination? Can the genome scan results already available and next-generation sequencing data be used to determine QTN? Should QTN be treated differently than markers at linkage disequilibrium with QTL in genetic evaluation programs? Data obtained by genome-wide association studies can be used to deduce QTL genotypes of sires via application of the a posteriori granddaughter design for concordance testing of putative QTN. This, together with next-generation sequencing technology, will dramatically reduce costs for QTN determination. By complete genome sequencing of 21 sires with many artificial insemination sons, it should be possible to determine concordance for all potential QTN, thus establishing the field of QTNomics. PMID:21338774

  10. A Simple Accurate Alternative to the Minimum-Deviation Method for the Determination of the Refractive Index of a Prism.

    ERIC Educational Resources Information Center

    Waldenstrom, S.; Naqvi, K. Razi

    1978-01-01

    Proposes an alternative to the classical minimum-deviation method for determining the refractive index of a prism. This new "fixed angle of incidence method" may find applications in research. (Author/GA)

  11. Quantitative analysis of electrophoresis data: novel curve fitting methodology and its application to the determination of a protein-DNA binding constant.

    PubMed

    Shadle, S E; Allen, D F; Guo, H; Pogozelski, W K; Bashkin, J S; Tullius, T D

    1997-02-15

    A computer program, GelExplorer, which uses a new methodology for obtaining quantitative information about electrophoresis has been developed. It provides a straightforward, easy-to-use graphical interface, and includes a number of features which offer significant advantages over existing methods for quantitative gel analysis. The method uses curve fitting with a nonlinear least-squares optimization to deconvolute overlapping bands. Unlike most curve fitting approaches, the data is treated in two dimensions, fitting all the data across the entire width of the lane. This allows for accurate determination of the intensities of individual, overlapping bands, and in particular allows imperfectly shaped bands to be accurately modeled. Experiments described in this paper demonstrate empirically that the Lorentzian lineshape reproduces the contours of an individual gel band and provides a better model than the Gaussian function for curve fitting of electrophoresis bands. Results from several fitting applications are presented and a discussion of the sources and magnitudes of uncertainties in the results is included. Finally, the method is applied to the quantitative analysis of a hydroxyl radical footprint titration experiment to obtain the free energy of binding of the lambda repressor protein to the OR1 operator DNA sequence.

  12. Near-infrared spectroscopy quantitative determination of Pefloxacin mesylate concentration in pharmaceuticals by using partial least squares and principal component regression multivariate calibration

    NASA Astrophysics Data System (ADS)

    Xie, Yunfei; Song, Yan; Zhang, Yong; Zhao, Bing

    2010-05-01

    Pefloxacin mesylate, a broad-spectrum antibacterial fluoroquinolone, has been widely used in clinical practice. Therefore, it is very important to detect the concentration of Pefloxacin mesylate. In this research, the near-infrared spectroscopy (NIRS) has been applied to quantitatively analyze on 108 injection samples, which was divided into a calibration set containing 89 samples and a prediction set containing 19 samples randomly. In order to get a satisfying result, partial least square (PLS) regression and principal components regression (PCR) have been utilized to establish quantitative models. Also, the process of establishing the models, parameters of the models, and prediction results were discussed in detail. In the PLS regression, the values of the coefficient of determination ( R2) and root mean square error of cross-validation (RMSECV) of PLS regression are 0.9263 and 0.00119, respectively. For comparison, though applying PCR method to get the values of R2 and RMSECV we obtained are 0.9685 and 0.00108, respectively. And the values of the standard error of prediction set (SEP) of PLS and PCR models are 0.001480 and 0.001140. The result of the prediction set suggests that these two quantitative analysis models have excellent generalization ability and prediction precision. However, for this PFLX injection samples, the PCR quantitative analysis model achieved more accurate results than the PLS model. The experimental results showed that NIRS together with PCR method provide rapid and accurate quantitative analysis of PFLX injection samples. Moreover, this study supplied technical support for the further analysis of other injection samples in pharmaceuticals.

  13. iMet-Q: A User-Friendly Tool for Label-Free Metabolomics Quantitation Using Dynamic Peak-Width Determination.

    PubMed

    Chang, Hui-Yin; Chen, Ching-Tai; Lih, T Mamie; Lynn, Ke-Shiuan; Juo, Chiun-Gung; Hsu, Wen-Lian; Sung, Ting-Yi

    2016-01-01

    Efficient and accurate quantitation of metabolites from LC-MS data has become an important topic. Here we present an automated tool, called iMet-Q (intelligent Metabolomic Quantitation), for label-free metabolomics quantitation from high-throughput MS1 data. By performing peak detection and peak alignment, iMet-Q provides a summary of quantitation results and reports ion abundance at both replicate level and sample level. Furthermore, it gives the charge states and isotope ratios of detected metabolite peaks to facilitate metabolite identification. An in-house standard mixture and a public Arabidopsis metabolome data set were analyzed by iMet-Q. Three public quantitation tools, including XCMS, MetAlign, and MZmine 2, were used for performance comparison. From the mixture data set, seven standard metabolites were detected by the four quantitation tools, for which iMet-Q had a smaller quantitation error of 12% in both profile and centroid data sets. Our tool also correctly determined the charge states of seven standard metabolites. By searching the mass values for those standard metabolites against Human Metabolome Database, we obtained a total of 183 metabolite candidates. With the isotope ratios calculated by iMet-Q, 49% (89 out of 183) metabolite candidates were filtered out. From the public Arabidopsis data set reported with two internal standards and 167 elucidated metabolites, iMet-Q detected all of the peaks corresponding to the internal standards and 167 metabolites. Meanwhile, our tool had small abundance variation (≤ 0.19) when quantifying the two internal standards and had higher abundance correlation (≥ 0.92) when quantifying the 167 metabolites. iMet-Q provides user-friendly interfaces and is publicly available for download at http://ms.iis.sinica.edu.tw/comics/Software_iMet-Q.html.

  14. iMet-Q: A User-Friendly Tool for Label-Free Metabolomics Quantitation Using Dynamic Peak-Width Determination

    PubMed Central

    Chang, Hui-Yin; Chen, Ching-Tai; Lih, T. Mamie; Lynn, Ke-Shiuan; Juo, Chiun-Gung; Hsu, Wen-Lian; Sung, Ting-Yi

    2016-01-01

    Efficient and accurate quantitation of metabolites from LC-MS data has become an important topic. Here we present an automated tool, called iMet-Q (intelligent Metabolomic Quantitation), for label-free metabolomics quantitation from high-throughput MS1 data. By performing peak detection and peak alignment, iMet-Q provides a summary of quantitation results and reports ion abundance at both replicate level and sample level. Furthermore, it gives the charge states and isotope ratios of detected metabolite peaks to facilitate metabolite identification. An in-house standard mixture and a public Arabidopsis metabolome data set were analyzed by iMet-Q. Three public quantitation tools, including XCMS, MetAlign, and MZmine 2, were used for performance comparison. From the mixture data set, seven standard metabolites were detected by the four quantitation tools, for which iMet-Q had a smaller quantitation error of 12% in both profile and centroid data sets. Our tool also correctly determined the charge states of seven standard metabolites. By searching the mass values for those standard metabolites against Human Metabolome Database, we obtained a total of 183 metabolite candidates. With the isotope ratios calculated by iMet-Q, 49% (89 out of 183) metabolite candidates were filtered out. From the public Arabidopsis data set reported with two internal standards and 167 elucidated metabolites, iMet-Q detected all of the peaks corresponding to the internal standards and 167 metabolites. Meanwhile, our tool had small abundance variation (≤0.19) when quantifying the two internal standards and had higher abundance correlation (≥0.92) when quantifying the 167 metabolites. iMet-Q provides user-friendly interfaces and is publicly available for download at http://ms.iis.sinica.edu.tw/comics/Software_iMet-Q.html. PMID:26784691

  15. iMet-Q: A User-Friendly Tool for Label-Free Metabolomics Quantitation Using Dynamic Peak-Width Determination.

    PubMed

    Chang, Hui-Yin; Chen, Ching-Tai; Lih, T Mamie; Lynn, Ke-Shiuan; Juo, Chiun-Gung; Hsu, Wen-Lian; Sung, Ting-Yi

    2016-01-01

    Efficient and accurate quantitation of metabolites from LC-MS data has become an important topic. Here we present an automated tool, called iMet-Q (intelligent Metabolomic Quantitation), for label-free metabolomics quantitation from high-throughput MS1 data. By performing peak detection and peak alignment, iMet-Q provides a summary of quantitation results and reports ion abundance at both replicate level and sample level. Furthermore, it gives the charge states and isotope ratios of detected metabolite peaks to facilitate metabolite identification. An in-house standard mixture and a public Arabidopsis metabolome data set were analyzed by iMet-Q. Three public quantitation tools, including XCMS, MetAlign, and MZmine 2, were used for performance comparison. From the mixture data set, seven standard metabolites were detected by the four quantitation tools, for which iMet-Q had a smaller quantitation error of 12% in both profile and centroid data sets. Our tool also correctly determined the charge states of seven standard metabolites. By searching the mass values for those standard metabolites against Human Metabolome Database, we obtained a total of 183 metabolite candidates. With the isotope ratios calculated by iMet-Q, 49% (89 out of 183) metabolite candidates were filtered out. From the public Arabidopsis data set reported with two internal standards and 167 elucidated metabolites, iMet-Q detected all of the peaks corresponding to the internal standards and 167 metabolites. Meanwhile, our tool had small abundance variation (≤ 0.19) when quantifying the two internal standards and had higher abundance correlation (≥ 0.92) when quantifying the 167 metabolites. iMet-Q provides user-friendly interfaces and is publicly available for download at http://ms.iis.sinica.edu.tw/comics/Software_iMet-Q.html. PMID:26784691

  16. Can we manage groundwater? A method to determine the quantitative testability of groundwater management plans

    NASA Astrophysics Data System (ADS)

    White, E. K.; Peterson, T. J.; Costelloe, J.; Western, A. W.; Carrara, E.

    2016-06-01

    Groundwater is the world's largest freshwater resource and due to overextraction, levels have declined in many regions causing extensive social and environmental impacts. Groundwater management seeks to balance and mitigate the detrimental impacts of development, with plans commonly used to outline management pathways. Thus, plan efficiency is crucial, but seldom are plans systematically and quantitatively assessed for effectiveness. This study frames groundwater management as a system control problem in order to develop a novel testability assessment rubric to determine if plans meet the requirements of a control loop, and subsequently, whether they can be quantitatively tested. Seven components of a management plan equivalent to basic components of a control loop were determined, and requirements of each component necessary to enable testability were defined. Each component was weighted based upon proposed relative importance, then segmented into rated categories depending on the degree the requirements were met. Component importance varied but, a defined objective or acceptable impact was necessary for plans to be testable. The rubric was developed within the context of the Australian groundwater management industry, and while use of the rubric is not limited to Australia, it was applied to 15 Australian groundwater management plans and approximately 47% were found to be testable. Considering the importance of effective groundwater management, and the central role of plans, our lack of ability to test many plans is concerning.

  17. Selective DNA Pooling for Determination of Linkage between a Molecular Marker and a Quantitative Trait Locus

    PubMed Central

    Darvasi, A.; Soller, M.

    1994-01-01

    Selective genotyping is a method to reduce costs in marker-quantitative trait locus (QTL) linkage determination by genotyping only those individuals with extreme, and hence most informative, quantitative trait values. The DNA pooling strategy (termed: ``selective DNA pooling'') takes this one step further by pooling DNA from the selected individuals at each of the two phenotypic extremes, and basing the test for linkage on marker allele frequencies as estimated from the pooled samples only. This can reduce genotyping costs of marker-QTL linkage determination by up to two orders of magnitude. Theoretical analysis of selective DNA pooling shows that for experiments involving backcross, F(2) and half-sib designs, the power of selective DNA pooling for detecting genes with large effect, can be the same as that obtained by individual selective genotyping. Power for detecting genes with small effect, however, was found to decrease strongly with increase in the technical error of estimating allele frequencies in the pooled samples. The effect of technical error, however, can be markedly reduced by replication of technical procedures. It is also shown that a proportion selected of 0.1 at each tail will be appropriate for a wide range of experimental conditions. PMID:7896115

  18. Quantitative dissection and stoichiometry determination of the human SET1/MLL histone methyltransferase complexes.

    PubMed

    van Nuland, Rick; Smits, Arne H; Pallaki, Paschalina; Jansen, Pascal W T C; Vermeulen, Michiel; Timmers, H T Marc

    2013-05-01

    Methylation of lysine 4 on histone H3 (H3K4) at promoters is tightly linked to transcriptional regulation in human cells. At least six different COMPASS-like multisubunit (SET1/MLL) complexes that contain methyltransferase activity for H3K4 have been described, but a comprehensive and quantitative analysis of these SET1/MLL complexes is lacking. We applied label-free quantitative mass spectrometry to determine the subunit composition and stoichiometry of the human SET1/MLL complexes. We identified both known and novel, unique and shared interactors and determined their distribution and stoichiometry over the different SET1/MLL complexes. In addition to being a core COMPASS subunit, the Dpy30 protein is a genuine subunit of the NURF chromatin remodeling complex. Furthermore, we identified the Bod1 protein as a discriminator between the SET1B and SET1A complexes, and we show that the H3K36me-interactor Psip1 preferentially binds to the MLL2 complex. Finally, absolute protein quantification in crude lysates mirrors many of the observed SET1/MLL complex stoichiometries. Our findings provide a molecular framework for understanding the diversity and abundance of the different SET1/MLL complexes, which together establish the H3K4 methylation landscape in human cells. PMID:23508102

  19. Quantitative determination of glyphosate in human serum by 1H NMR spectroscopy.

    PubMed

    Cartigny, Bernard; Azaroual, Nathalie; Imbenotte, Michel; Mathieu, Daniel; Parmentier, Erika; Vermeersch, Gaston; Lhermitte, Michel

    2008-01-15

    The determination and quantification of glyphosate in serum using (1)H NMR spectroscopy is reported. This method permitted serum samples to be analysed without derivatization or any other sample pre-treatment, using 3-trimethylsilyl 2,2',3,3'-tetradeuteropropionic acid (TSP-d(4)) as a qualitative and quantitative standard. Characterization of the herbicide N-(phosphonomethyl)glycine was performed by analysing chemical shifts and coupling constant patterns. Quantification was performed by relative integration of CH(2)-P protons to the TSP-d(4) resonance peak. The method was tested for repeatability (n=5) and yielded coefficients of variation of 1% and 3%, respectively: detection and quantification limits were also determined and were 0.03 and 0.1mmol/L, respectively. The method was applied to the quantification of glyphosate in a case of acute poisoning. PMID:18371753

  20. Communication: Vibrationally resolved photoelectron spectroscopy of the tetracyanoquinodimethane (TCNQ) anion and accurate determination of the electron affinity of TCNQ

    SciTech Connect

    Zhu, Guo-Zhu; Wang, Lai-Sheng

    2015-12-14

    Tetracyanoquinodimethane (TCNQ) is widely used as an electron acceptor to form highly conducting organic charge-transfer solids. Surprisingly, the electron affinity (EA) of TCNQ is not well known and has never been directly measured. Here, we report vibrationally resolved photoelectron spectroscopy (PES) of the TCNQ{sup −} anion produced using electrospray and cooled in a cryogenic ion trap. Photoelectron spectrum taken at 354.7 nm represents the detachment transition from the ground state of TCNQ{sup −} to that of neutral TCNQ with a short vibrational progression. The EA of TCNQ is measured accurately to be 3.383 ± 0.001 eV (27 289 ± 8 cm{sup −1}), compared to the 2.8 ± 0.1 eV value known in the literature and measured previously using collisional ionization technique. In addition, six vibrational peaks are observed in the photoelectron spectrum, yielding vibrational frequencies for three totally symmetric modes of TCNQ. Two-photon PES via a bound electronic excited state of TCNQ{sup −} at 3.100 eV yields a broad low kinetic energy peak due to fast internal conversion to vibrationally excited levels of the anion ground electronic state. The high EA measured for TCNQ underlies its ability as a good electron acceptor.

  1. Communication: Vibrationally resolved photoelectron spectroscopy of the tetracyanoquinodimethane (TCNQ) anion and accurate determination of the electron affinity of TCNQ

    NASA Astrophysics Data System (ADS)

    Zhu, Guo-Zhu; Wang, Lai-Sheng

    2015-12-01

    Tetracyanoquinodimethane (TCNQ) is widely used as an electron acceptor to form highly conducting organic charge-transfer solids. Surprisingly, the electron affinity (EA) of TCNQ is not well known and has never been directly measured. Here, we report vibrationally resolved photoelectron spectroscopy (PES) of the TCNQ- anion produced using electrospray and cooled in a cryogenic ion trap. Photoelectron spectrum taken at 354.7 nm represents the detachment transition from the ground state of TCNQ- to that of neutral TCNQ with a short vibrational progression. The EA of TCNQ is measured accurately to be 3.383 ± 0.001 eV (27 289 ± 8 cm-1), compared to the 2.8 ± 0.1 eV value known in the literature and measured previously using collisional ionization technique. In addition, six vibrational peaks are observed in the photoelectron spectrum, yielding vibrational frequencies for three totally symmetric modes of TCNQ. Two-photon PES via a bound electronic excited state of TCNQ- at 3.100 eV yields a broad low kinetic energy peak due to fast internal conversion to vibrationally excited levels of the anion ground electronic state. The high EA measured for TCNQ underlies its ability as a good electron acceptor.

  2. Quantitative profiling of bile acids in biofluids and tissues based on accurate mass high resolution LC-FT-MS: compound class targeting in a metabolomics workflow.

    PubMed

    Bobeldijk, Ivana; Hekman, Maarten; de Vries-van der Weij, Jitske; Coulier, Leon; Ramaker, Raymond; Kleemann, Robert; Kooistra, Teake; Rubingh, Carina; Freidig, Andreas; Verheij, Elwin

    2008-08-15

    We report a sensitive, generic method for quantitative profiling of bile acids and other endogenous metabolites in small quantities of various biological fluids and tissues. The method is based on a straightforward sample preparation, separation by reversed-phase high performance liquid-chromatography mass spectrometry (HPLC-MS) and electrospray ionisation in the negative ionisation mode (ESI-). Detection is performed in full scan using the linear ion trap Fourier transform mass spectrometer (LTQ-FTMS) generating data for many (endogenous) metabolites, not only bile acids. A validation of the method in urine, plasma and liver was performed for 17 bile acids including their taurine, sulfate and glycine conjugates. The method is linear in the 0.01-1 microM range. The accuracy in human plasma ranges from 74 to 113%, in human urine 77 to 104% and in mouse liver 79 to 140%. The precision ranges from 2 to 20% for pooled samples even in studies with large number of samples (n>250). The method was successfully applied to a multi-compartmental APOE*3-Leiden mouse study, the main goal of which was to analyze the effect of increasing dietary cholesterol concentrations on hepatic cholesterol homeostasis and bile acid synthesis. Serum and liver samples from different treatment groups were profiled with the new method. Statistically significant differences between the diet groups were observed regarding total as well as individual bile acid concentrations.

  3. Influence of storage time on DNA of Chlamydia trachomatis, Ureaplasma urealyticum, and Neisseria gonorrhoeae for accurate detection by quantitative real-time polymerase chain reaction.

    PubMed

    Lu, Y; Rong, C Z; Zhao, J Y; Lao, X J; Xie, L; Li, S; Qin, X

    2016-01-01

    The shipment and storage conditions of clinical samples pose a major challenge to the detection accuracy of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) when using quantitative real-time polymerase chain reaction (qRT-PCR). The aim of the present study was to explore the influence of storage time at 4°C on the DNA of these pathogens and its effect on their detection by qRT-PCR. CT, NG, and UU positive genital swabs from 70 patients were collected, and DNA of all samples were extracted and divided into eight aliquots. One aliquot was immediately analyzed with qRT-PCR to assess the initial pathogen load, whereas the remaining samples were stored at 4°C and analyzed after 1, 2, 3, 7, 14, 21, and 28 days. No significant differences in CT, NG, and UU DNA loads were observed between baseline (day 0) and the subsequent time points (days 1, 2, 3, 7, 14, 21, and 28) in any of the 70 samples. Although a slight increase in DNA levels was observed at day 28 compared to day 0, paired sample t-test results revealed no significant differences between the mean DNA levels at different time points following storage at 4°C (all P>0.05). Overall, the CT, UU, and NG DNA loads from all genital swab samples were stable at 4°C over a 28-day period. PMID:27580005

  4. Influence of storage time on DNA of Chlamydia trachomatis, Ureaplasma urealyticum, and Neisseria gonorrhoeae for accurate detection by quantitative real-time polymerase chain reaction.

    PubMed

    Lu, Y; Rong, C Z; Zhao, J Y; Lao, X J; Xie, L; Li, S; Qin, X

    2016-08-25

    The shipment and storage conditions of clinical samples pose a major challenge to the detection accuracy of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) when using quantitative real-time polymerase chain reaction (qRT-PCR). The aim of the present study was to explore the influence of storage time at 4°C on the DNA of these pathogens and its effect on their detection by qRT-PCR. CT, NG, and UU positive genital swabs from 70 patients were collected, and DNA of all samples were extracted and divided into eight aliquots. One aliquot was immediately analyzed with qRT-PCR to assess the initial pathogen load, whereas the remaining samples were stored at 4°C and analyzed after 1, 2, 3, 7, 14, 21, and 28 days. No significant differences in CT, NG, and UU DNA loads were observed between baseline (day 0) and the subsequent time points (days 1, 2, 3, 7, 14, 21, and 28) in any of the 70 samples. Although a slight increase in DNA levels was observed at day 28 compared to day 0, paired sample t-test results revealed no significant differences between the mean DNA levels at different time points following storage at 4°C (all P>0.05). Overall, the CT, UU, and NG DNA loads from all genital swab samples were stable at 4°C over a 28-day period.

  5. Influence of storage time on DNA of Chlamydia trachomatis, Ureaplasma urealyticum, and Neisseria gonorrhoeae for accurate detection by quantitative real-time polymerase chain reaction

    PubMed Central

    Lu, Y.; Rong, C.Z.; Zhao, J.Y.; Lao, X.J.; Xie, L.; Li, S.; Qin, X.

    2016-01-01

    The shipment and storage conditions of clinical samples pose a major challenge to the detection accuracy of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) when using quantitative real-time polymerase chain reaction (qRT-PCR). The aim of the present study was to explore the influence of storage time at 4°C on the DNA of these pathogens and its effect on their detection by qRT-PCR. CT, NG, and UU positive genital swabs from 70 patients were collected, and DNA of all samples were extracted and divided into eight aliquots. One aliquot was immediately analyzed with qRT-PCR to assess the initial pathogen load, whereas the remaining samples were stored at 4°C and analyzed after 1, 2, 3, 7, 14, 21, and 28 days. No significant differences in CT, NG, and UU DNA loads were observed between baseline (day 0) and the subsequent time points (days 1, 2, 3, 7, 14, 21, and 28) in any of the 70 samples. Although a slight increase in DNA levels was observed at day 28 compared to day 0, paired sample t-test results revealed no significant differences between the mean DNA levels at different time points following storage at 4°C (all P>0.05). Overall, the CT, UU, and NG DNA loads from all genital swab samples were stable at 4°C over a 28-day period. PMID:27580005

  6. Quantitative determination of calcium, magnesium, and zinc in fingernails by laser-induced breakdown spectroscopy.

    PubMed

    Rusak, David A; Zeleniak, Ann E; Obuhosky, Jillian L; Holdren, Scott M; Noldy, Craig A

    2013-12-15

    Quantitative determination of Ca, Mg, and Zn in fingernails was performed with laser-induced breakdown spectroscopy. Two different methods of producing solid standards for calibration were explored - preparation of keratin pellets and deposition of aqueous solutions to filter papers. Measurements of the temperature and electron density of the plasma produced on keratin pellets, filter paper, and nails were performed, and it was determined that the standards prepared on filter paper gave plasma temperatures and electron densities closer to those observed on the nails. The ablation rate of the filter paper was also more similar to that of the nails. Using calibration curves produced using these filter paper standards, Ca, Mg, and Zn were determined in fingernails of 11 subjects. For comparison, the same samples were digested and atomic absorption was used to determine the same three elements. The differences in results are discussed in light of sample homogeneity and instrumental precision; the best agreement was obtained for determination of Zn. The work suggests that the filter paper method of standard preparation may be appropriate for LIBS analysis of other samples that give relatively low temperature, low electron density plasmas (i.e., polymers).

  7. Noninvasive determination of bone stiffness for distraction osteogenesis by quantitative computed tomography scans.

    PubMed

    Harp, J H; Aronson, J; Hollis, M

    1994-04-01

    Apparent density of canine tibial specimens was correlated to measured quantitative computed tomography (QCT) numbers, and a conversion equation was derived. Finite element analysis models of canine tibiae were constructed using spatial coordinates of converted QCT data. Two QCT-derived finite elemental analysis models were constructed for a pair of matched tibiae: one lengthened 15% by distraction osteogenesis and one contralateral control. Two equations relating the modulus of elasticity to apparent density were used. These models were then verified mechanically by measuring strains of the two specimens under applied load. Equation I correlated best with measured strains in the less-dense regenerate bone of the distracted tibia (R2 = 0.914, p = 0.0028, n = 6); Equation II performed best in the denser control tibia (R2 = 0.820, p = 0.0129, n = 6). These results demonstrate that the stiffness of tubular bones, including cortical and heterogenous cancellous structure, can be accurately predicted by a noninvasive QCT scan.

  8. Light fluence correction for quantitative determination of tissue absorption coefficient using multi-spectral optoacoustic tomography

    NASA Astrophysics Data System (ADS)

    Brochu, Frederic M.; Joseph, James; Tomaszewski, Michal; Bohndiek, Sarah E.

    2015-07-01

    MultiSpectral Optoacoustic Tomography (MSOT) is a fast developing imaging modality, combining the high resolution and penetration depth of ultrasound with the excellent contrast from optical imaging of tissue. Absorption and scattering of the near infrared excitation light modulates the spectral profile of light as it propagates deep into biological tissue, meaning the images obtained provide only qualitative insight into the distribution of tissue chromophores. The goal of this work is to accurately recover the spectral profile of excitation light by modelling light fluence in the data reconstruction, to enable quantitative imaging. We worked with a commercial small animal MSOT scanner and developed our light fluence correction for its' cylindrical geometry. Optoacoustic image reconstruction pinpoints the sources of acoustic waves detected by the transducers and returns the initial pressure amplitude at these points. This pressure is the product of the dimensionless Grüneisen parameter, the absorption coefficient and the light fluence. Under the condition of constant Grüneisen parameter and well modelled light fluence, there is a linear relationship between the initial pressure amplitude measured in the optoacoustic image and the absorption coefficient. We were able to reproduce this linear relationship in different physical regions of an agarose gel phantom containing targets of known optical absorption coefficient, demonstrating that our light fluence model was working. We also demonstrate promising results of light fluence correction effects on in vivo data.

  9. 45 CFR 261.64 - How will we determine whether a State's work verification procedures ensure an accurate work...

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... interim Work Verification Plan by September 30, 2006 and a complete Work Verification Plan that we approved by September 30, 2007. (b) A “complete” Work Verification Plan means that: (1) The plan includes... 45 Public Welfare 2 2010-10-01 2010-10-01 false How will we determine whether a State's...

  10. Determination of accurate electron chiral asymmetries in fenchone and camphor in the VUV range: sensitivity to isomerism and enantiomeric purity.

    PubMed

    Nahon, Laurent; Nag, Lipsa; Garcia, Gustavo A; Myrgorodska, Iuliia; Meierhenrich, Uwe; Beaulieu, Samuel; Wanie, Vincent; Blanchet, Valérie; Géneaux, Romain; Powis, Ivan

    2016-05-14

    Photoelectron circular dichroism (PECD) manifests itself as an intense forward/backward asymmetry in the angular distribution of photoelectrons produced from randomly-oriented enantiomers by photoionization with circularly-polarized light (CPL). As a sensitive probe of both photoionization dynamics and of the chiral molecular potential, PECD attracts much interest especially with the recent performance of related experiments with visible and VUV laser sources. Here we report, by use of quasi-perfect CPL VUV synchrotron radiation and using a double imaging photoelectron/photoion coincidence (i(2)PEPICO) spectrometer, new and very accurate values of the corresponding asymmetries on showcase chiral isomers: camphor and fenchone. These data have additionally been normalized to the absolute enantiopurity of the sample as measured by a chromatographic technique. They can therefore be used as benchmarking data for new PECD experiments, as well as for theoretical models. In particular we found, especially for the outermost orbital of both molecules, a good agreement with CMS-Xα PECD modeling over the whole VUV range. We also report a spectacular sensitivity of PECD to isomerism for slow electrons, showing large and opposite asymmetries when comparing R-camphor to R-fenchone (respectively -10% and +16% around 10 eV). In the course of this study, we could also assess the analytical potential of PECD. Indeed, the accuracy of the data we provide are such that limited departure from perfect enantiopurity in the sample we purchased could be detected and estimated in excellent agreement with the analysis performed in parallel via a chromatographic technique, establishing a new standard of accuracy, in the ±1% range, for enantiomeric excess measurement via PECD. The i(2)PEPICO technique allows correlating PECD measurements to specific parent ion masses, which would allow its application to analysis of complex mixtures. PMID:27095534

  11. Validation of reference genes for accurate normalization of gene expression for real time-quantitative PCR in strawberry fruits using different cultivars and osmotic stresses.

    PubMed

    Galli, Vanessa; Borowski, Joyce Moura; Perin, Ellen Cristina; Messias, Rafael da Silva; Labonde, Julia; Pereira, Ivan dos Santos; Silva, Sérgio Delmar Dos Anjos; Rombaldi, Cesar Valmor

    2015-01-10

    The increasing demand of strawberry (Fragaria×ananassa Duch) fruits is associated mainly with their sensorial characteristics and the content of antioxidant compounds. Nevertheless, the strawberry production has been hampered due to its sensitivity to abiotic stresses. Therefore, to understand the molecular mechanisms highlighting stress response is of great importance to enable genetic engineering approaches aiming to improve strawberry tolerance. However, the study of expression of genes in strawberry requires the use of suitable reference genes. In the present study, seven traditional and novel candidate reference genes were evaluated for transcript normalization in fruits of ten strawberry cultivars and two abiotic stresses, using RefFinder, which integrates the four major currently available software programs: geNorm, NormFinder, BestKeeper and the comparative delta-Ct method. The results indicate that the expression stability is dependent on the experimental conditions. The candidate reference gene DBP (DNA binding protein) was considered the most suitable to normalize expression data in samples of strawberry cultivars and under drought stress condition, and the candidate reference gene HISTH4 (histone H4) was the most stable under osmotic stresses and salt stress. The traditional genes GAPDH (glyceraldehyde-3-phosphate dehydrogenase) and 18S (18S ribosomal RNA) were considered the most unstable genes in all conditions. The expression of phenylalanine ammonia lyase (PAL) and 9-cis epoxycarotenoid dioxygenase (NCED1) genes were used to further confirm the validated candidate reference genes, showing that the use of an inappropriate reference gene may induce erroneous results. This study is the first survey on the stability of reference genes in strawberry cultivars and osmotic stresses and provides guidelines to obtain more accurate RT-qPCR results for future breeding efforts.

  12. Accurate determination of electronic transport properties of silicon wafers by nonlinear photocarrier radiometry with multiple pump beam sizes

    SciTech Connect

    Wang, Qian; Li, Bincheng

    2015-12-07

    In this paper, photocarrier radiometry (PCR) technique with multiple pump beam sizes is employed to determine simultaneously the electronic transport parameters (the carrier lifetime, the carrier diffusion coefficient, and the front surface recombination velocity) of silicon wafers. By employing the multiple pump beam sizes, the influence of instrumental frequency response on the multi-parameter estimation is totally eliminated. A nonlinear PCR model is developed to interpret the PCR signal. Theoretical simulations are performed to investigate the uncertainties of the estimated parameter values by investigating the dependence of a mean square variance on the corresponding transport parameters and compared to that obtained by the conventional frequency-scan method, in which only the frequency dependences of the PCR amplitude and phase are recorded at single pump beam size. Simulation results show that the proposed multiple-pump-beam-size method can improve significantly the accuracy of the determination of the electronic transport parameters. Comparative experiments with a p-type silicon wafer with resistivity 0.1–0.2 Ω·cm are performed, and the electronic transport properties are determined simultaneously. The estimated uncertainties of the carrier lifetime, diffusion coefficient, and front surface recombination velocity are approximately ±10.7%, ±8.6%, and ±35.4% by the proposed multiple-pump-beam-size method, which is much improved than ±15.9%, ±29.1%, and >±50% by the conventional frequency-scan method. The transport parameters determined by the proposed multiple-pump-beam-size PCR method are in good agreement with that obtained by a steady-state PCR imaging technique.

  13. Combination of Eight Alleles at Four Quantitative Trait Loci Determines Grain Length in Rice.

    PubMed

    Zeng, Yuxiang; Ji, Zhijuan; Wen, Zhihua; Liang, Yan; Yang, Changdeng

    2016-01-01

    Grain length is an important quantitative trait in rice (Oryza sativa L.) that influences both grain yield and exterior quality. Although many quantitative trait loci (QTLs) for grain length have been identified, it is still unclear how different alleles from different QTLs regulate grain length coordinately. To explore the mechanisms of QTL combination in the determination of grain length, five mapping populations, including two F2 populations, an F3 population, an F7 recombinant inbred line (RIL) population, and an F8 RIL population, were developed from the cross between the U.S. tropical japonica variety 'Lemont' and the Chinese indica variety 'Yangdao 4' and grown under different environmental conditions. Four QTLs (qGL-3-1, qGL-3-2, qGL-4, and qGL-7) for grain length were detected using both composite interval mapping and multiple interval mapping methods in the mapping populations. In each locus, there was an allele from one parent that increased grain length and another allele from another parent that decreased it. The eight alleles in the four QTLs were analyzed to determine whether these alleles act additively across loci, and lead to a linear relationship between the predicted breeding value of QTLs and phenotype. Linear regression analysis suggested that the combination of eight alleles determined grain length. Plants carrying more grain length-increasing alleles had longer grain length than those carrying more grain length-decreasing alleles. This trend was consistent in all five mapping populations and demonstrated the regulation of grain length by the four QTLs. Thus, these QTLs are ideal resources for modifying grain length in rice.

  14. High-throughput double quantitative competitive polymerase chain reaction for determination of genetically modified organisms.

    PubMed

    Mavropoulou, Anastasia K; Koraki, Theodora; Ioannou, Penelope C; Christopoulos, Theodore K

    2005-08-01

    Quantitative competitive polymerase chain reaction (PCR), especially the double competitive PCR methods (DC-PCR), have evolved as reliable approaches to quantification of genetically modified organisms (GMO) in food. However, DC-PCR is a low-throughput method because it requires titration of each sample with various amounts of a competitive internal standard, a protocol that involves several PCRs per sample followed by electrophoresis and densitometry. To address this drawback, we have developed a new method for GMO quantification, namely, a high-throughput double quantitative competitive PCR (HT-DCPCR). In HT-DCPCR, electrophoresis and densitometry are replaced by a rapid, microtiter well-based bioluminometric hybridization assay and there is no need for titration of each sample. The determination of GM soya was chosen as a model. We have constructed internal standards (DNA competitors) both for the 35S promoter sequence and for a plant-specific reference gene (lectin). The competitors have identical size and share the same primer binding sites with the target sequences but differ in a 24-bp internal segment. Each target sequence (35S and lectin) is coamplified with a constant amount (1000 copies) of the respective competitor. The four amplified fragments are hybridized with specific probes and captured on a universal solid phase to achieve simplicity and high throughput. The hybrids are determined by using streptavidin conjugated to the photoprotein aequorin. The ratio of the luminescence values obtained for the target and the competitor is linearly related to the starting amount of target DNA. The limit of quantification for the 35S promoter is 24 copies. The proposed method was evaluated by determining the GMO content of soybean powder certified reference materials. Also HT-DCPCR was compared to real-time PCR in a variety of real samples.

  15. Evaluation of reference genes for accurate normalization of gene expression for real time-quantitative PCR in Pyrus pyrifolia using different tissue samples and seasonal conditions.

    PubMed

    Imai, Tsuyoshi; Ubi, Benjamin E; Saito, Takanori; Moriguchi, Takaya

    2014-01-01

    We have evaluated suitable reference genes for real time (RT)-quantitative PCR (qPCR) analysis in Japanese pear (Pyrus pyrifolia). We tested most frequently used genes in the literature such as β-Tubulin, Histone H3, Actin, Elongation factor-1α, Glyceraldehyde-3-phosphate dehydrogenase, together with newly added genes Annexin, SAND and TIP41. A total of 17 primer combinations for these eight genes were evaluated using cDNAs synthesized from 16 tissue samples from four groups, namely: flower bud, flower organ, fruit flesh and fruit skin. Gene expression stabilities were analyzed using geNorm and NormFinder software packages or by ΔCt method. geNorm analysis indicated three best performing genes as being sufficient for reliable normalization of RT-qPCR data. Suitable reference genes were different among sample groups, suggesting the importance of validation of gene expression stability of reference genes in the samples of interest. Ranking of stability was basically similar between geNorm and NormFinder, suggesting usefulness of these programs based on different algorithms. ΔCt method suggested somewhat different results in some groups such as flower organ or fruit skin; though the overall results were in good correlation with geNorm or NormFinder. Gene expression of two cold-inducible genes PpCBF2 and PpCBF4 were quantified using the three most and the three least stable reference genes suggested by geNorm. Although normalized quantities were different between them, the relative quantities within a group of samples were similar even when the least stable reference genes were used. Our data suggested that using the geometric mean value of three reference genes for normalization is quite a reliable approach to evaluating gene expression by RT-qPCR. We propose that the initial evaluation of gene expression stability by ΔCt method, and subsequent evaluation by geNorm or NormFinder for limited number of superior gene candidates will be a practical way of finding out

  16. The role of cognitive switching in head-up displays. [to determine pilot ability to accurately extract information from either of two sources

    NASA Technical Reports Server (NTRS)

    Fischer, E.

    1979-01-01

    The pilot's ability to accurately extract information from either one or both of two superimposed sources of information was determined. Static, aerial, color 35 mm slides of external runway environments and slides of corresponding static head-up display (HUD) symbology were used as the sources. A three channel tachistoscope was utilized to show either the HUD alone, the scene alone, or the two slides superimposed. Cognitive performance of the pilots was assessed by determining the percentage of correct answers given to two HUD related questions, two scene related questions, or one HUD and one scene related question.

  17. A simple method for the accurate determination of the Henry's law constant for highly sorptive, semivolatile organic compounds.

    PubMed

    Kim, Yong-Hyun; Kim, Ki-Hyun

    2016-01-01

    A novel technique is developed to determine the Henry's law constants (HLCs) of seven volatile fatty acids (VFAs) with significantly high solubility using a combined application of thermal desorber/gas chromatography/mass spectrometry (TD/GC/MS). In light of the strong sorptive properties of these semi-volatile organic compounds (SVOCs), their HLCs were determined by properly evaluating the fraction lost on the surface of the materials used to induce equilibrium (vial, gas-tight syringe, and sorption tube). To this end, a total of nine repeated experiments were conducted in a closed (static) system at three different gas/liquid volume ratios. The best estimates for HLCs (M/atm) were thus 7,200 (propionic acid), 4,700 (i-butyric acid), 4,400 (n-butyric acid), 2,700 (i-valeric acid), 2,400 (n-valeric acid), 1,000 (hexanoic acid), and 1,500 (heptanoic acid). The differences in the HLC values between this study and previous studies, if assessed in terms of the percent difference, ranged from 9.2% (n-valeric acid) to 55.7% (i-valeric acid). We overcame the main cause of errors encountered in previous studies by performing the proper correction of the sorptive losses of the SVOCs that inevitably took place, particularly on the walls of the equilibration systems (mainly the headspace vial and/or the glass tight syringe). PMID:26577086

  18. Accurate determination of the Gibbs energy of Cu-Zr melts using the thermodynamic integration method in Monte Carlo simulations

    NASA Astrophysics Data System (ADS)

    Harvey, J.-P.; Gheribi, A. E.; Chartrand, P.

    2011-08-01

    The design of multicomponent alloys used in different applications based on specific thermo-physical properties determined experimentally or predicted from theoretical calculations is of major importance in many engineering applications. A procedure based on Monte Carlo simulations (MCS) and the thermodynamic integration (TI) method to improve the quality of the predicted thermodynamic properties calculated from classical thermodynamic calculations is presented in this study. The Gibbs energy function of the liquid phase of the Cu-Zr system at 1800 K has been determined based on this approach. The internal structure of Cu-Zr melts and amorphous alloys at different temperatures, as well as other physical properties were also obtained from MCS in which the phase trajectory was modeled by the modified embedded atom model formalism. A rigorous comparison between available experimental data and simulated thermo-physical properties obtained from our MCS is presented in this work. The modified quasichemical model in the pair approximation was parameterized using the internal structure data obtained from our MCS and the precise Gibbs energy function calculated at 1800 K from the TI method. The predicted activity of copper in Cu-Zr melts at 1499 K obtained from our thermodynamic optimization was corroborated by experimental data found in the literature. The validity of the amplitude of the entropy of mixing obtained from the in silico procedure presented in this work was analyzed based on the thermodynamic description of hard sphere mixtures.

  19. Quantitative Determination of Common Urinary Odorants and Their Glucuronide Conjugates in Human Urine

    PubMed Central

    Wagenstaller, Maria; Buettner, Andrea

    2013-01-01

    Our previous study on the identification of common odorants and their conjugates in human urine demonstrated that this substance fraction is a little-understood but nonetheless a promising medium for analysis and diagnostics in this easily accessible physiological medium. Smell as an indicator for diseases, or volatile excretion in the course of dietary processes bares high potential for a series of physiological insights. Still, little is known today about the quantitative composition of odorous or volatile targets, as well as their non-volatile conjugates, both with regard to their common occurrence in urine of healthy subjects, as well as in that of individuals suffering from diseases or other physiological misbalancing. Accordingly, the aim of our study was to develop a highly sensitive and selective approach to determine the common quantitative composition of selected odorant markers in healthy human subjects, as well as their corresponding glucuronide conjugates. We used one- and two-dimensional high resolution gas chromatography-mass spectrometry in combination with stable isotope dilution assays to quantify commonly occurring and potent odorants in human urine. The studies were carried out on both native urine and on urine that had been treated by glucuronidase assays, with analysis of the liberated odor-active compounds using the same techniques. Analytical data are discussed with regard to their potential translation as future diagnostic tool. PMID:24958143

  20. Qualitative and Quantitative Determination and Resistance Patterns of Salmonella from Poultry Carcasses.

    PubMed

    Yamatogi, R S; Oliveira, H C; Possebon, F S; Pantoja, J C F; Joaquim, J G F; Pinto, J P A N; Araújo, J P

    2016-06-01

    Poultry products are important in the transmission of zoonotic pathogens, mainly Salmonella. This genus causes millions of foodborne diseases worldwide every year. Cross-contamination by food sources in human cases of salmonellosis and the increase in resistant strains have become important issues. A qualitative and quantitative Salmonella detection method was utilized in a poultry slaughterhouse in São Paulo State, Brazil. We collected 33 samples from different batches of carcasses. Each sample was analyzed at three process points: postbleeding, postdefeathering, and postchilling. A fourth point, retail simulation, was added to simulate retail market storage at 5°C for 72 h. The qualitative methods revealed 100% (33 samples) contamination at postbleeding, 39% (13 samples) contamination at postdefeathering, 58% (19 samples) contamination at postchilling, and 30% (10 samples) contamination at the retail simulation. The quantitative results, determined by the most-probable-number (MPN) technique, ranged from <0.03 to >2,400 MPN/g. We identified 23 Salmonella serovars; the most prevalent were Mbandaka, Senftenberg, and Enteritidis. Resistance to nalidixic acid was significantly more common (P < 0.05) than resistance to other antimicrobial agents. Five multidrug-resistant strains were identified. This study contributes important epidemiological data and demonstrates the need to improve sanitary conditions in slaughterhouses.

  1. Determination of melengestrol acetate in feedstuffs with liquid chromatographic preparatory column cleanup and quantitative analysis.

    PubMed

    Weigand, J L; Dille, D S

    1988-01-01

    Melengestrol acetate (MGA) is determined by liquid chromatography using a fraction from preparatory LC as a means of sample cleanup for feedstuffs, both dry and liquid. Dry ground feed is Soxhlet extracted with hexane and passed through a 2% deactivated alumina column for initial cleanup. The eluate is evaporated, redissolved in methanol, filtered, and injected onto a preparatory LC column. The fraction containing MGA is separated from the remaining matrix, evaporated to dryness, dissolved in methanol, and quantitated by LC analysis. Liquid supplements are extracted in methanol, and the extract is evaporated to near dryness. The residue is diluted with water, extracted with chloroform, passed through sodium sulfate, and evaporated to dryness. The remaining sample is dissolved in methanol prior to preparative LC and quantitative LC. Recoveries for 2 laboratory-fortified commercial feeds, one dry and one liquid, containing 0.39 and 0.40 mg/lb, were 98.3% +/- 4.4 and 95.8% +/- 4.3, respectively. Results compare favorably with existing methods. Up to a 4-fold time savings was realized by this method without automation.

  2. Quantitative HPLC Analysis of an Analgesic/Caffeine Formulation: Determination of Caffeine

    NASA Astrophysics Data System (ADS)

    Ferguson, Glenda K.

    1998-04-01

    A modern high performance liquid chromatography (HPLC) laboratory experiment which entails the separation of acetaminophen, aspirin, and caffeine and the quantitative assay of caffeine in commercial mixtures of these compounds has been developed. Our HPLC protocol resolves these compounds in only three minutes with a straightforward chromatographic apparatus which consists of a C-18 column, an isocratic mobile phase, UV detection at 254 nm, and an integrator; an expensive, sophisticated system is not required. The separation is both repeatable and rapid. Moreover, the experiment can be completed in a single three-hour period. The experiment is appropriate for any chemistry student who has completed a minimum of one year of general chemistry and is ideal for an analytical or instrumental analysis course. The experiment detailed herein involves the determination of caffeine in Goody's Extra Strength Headache Powders, a commercially available medication which contains acetaminophen, aspirin, and caffeine as active ingredients. However, the separation scheme is not limited to this brand of medication nor is it limited to caffeine as the analyte. With only minor procedural modifications, students can simultaneously quantitate all of these compounds in a commercial mixture. In our procedure, students prepare a series of four caffeine standard solutions as well as a solution from a pharmaceutical analgesic/caffeine mixture, chromatographically analyze each solution in quadruplicate, and plot relative average caffeine standard peak area versus concentration. From the mathematical relationship that results, the concentration of caffeine in the commercial formulation is obtained. Finally, the absolute standard deviation of the mean concentration is calculated.

  3. Qualitative and Quantitative Determination and Resistance Patterns of Salmonella from Poultry Carcasses.

    PubMed

    Yamatogi, R S; Oliveira, H C; Possebon, F S; Pantoja, J C F; Joaquim, J G F; Pinto, J P A N; Araújo, J P

    2016-06-01

    Poultry products are important in the transmission of zoonotic pathogens, mainly Salmonella. This genus causes millions of foodborne diseases worldwide every year. Cross-contamination by food sources in human cases of salmonellosis and the increase in resistant strains have become important issues. A qualitative and quantitative Salmonella detection method was utilized in a poultry slaughterhouse in São Paulo State, Brazil. We collected 33 samples from different batches of carcasses. Each sample was analyzed at three process points: postbleeding, postdefeathering, and postchilling. A fourth point, retail simulation, was added to simulate retail market storage at 5°C for 72 h. The qualitative methods revealed 100% (33 samples) contamination at postbleeding, 39% (13 samples) contamination at postdefeathering, 58% (19 samples) contamination at postchilling, and 30% (10 samples) contamination at the retail simulation. The quantitative results, determined by the most-probable-number (MPN) technique, ranged from <0.03 to >2,400 MPN/g. We identified 23 Salmonella serovars; the most prevalent were Mbandaka, Senftenberg, and Enteritidis. Resistance to nalidixic acid was significantly more common (P < 0.05) than resistance to other antimicrobial agents. Five multidrug-resistant strains were identified. This study contributes important epidemiological data and demonstrates the need to improve sanitary conditions in slaughterhouses. PMID:27296598

  4. [Sulpiride poisoning--case report confirmed with the quantitative determination of the xenobiotic serum level].

    PubMed

    Ciszowski, Krzysztof; Szpak, Dorota; Wilimowska, Jolanta; Groszek, Barbara

    2011-01-01

    Despite above 40 years the presence of sulpride on the pharmaceutical market, the acute poisonings are poorly reported in the medical literature. The discussed case of sulpiride intoxication concerns ingestion probably dose of 12 g, that exceeded 10-fold maximum therapeutic dose. 16-year-old girl, with no previous sulpiride treatment, was admitted to the Toxicology Department about 3 hours after ingestion. In clinical picture she presented quantitative consciousness disturbances with maximum 10 scores in GCS scale, with tendency to low BP (minimum 88/45 mmHg) and episode of orthostatic hypotension. The ECG demonstrated: normogram, sinus tachycardia with a heart rate of 125 beats/min, PQ = 120 ms, QRS = 80 ms, prolongation of QTc to 519,6 ms and unspecific changes of ST-T syndrome. The qualitative toxicological test confirmed the presence of chlorprothixene in urine, but the serum therapeutic concentration (0.126 microg/ml) excluded the overdose. The quantitative determination of sulpiride serum concentration confirmed acute sulpiride poisoning. The measured sulpiride toxic concentration on admission and in the consecutive hours were from 13.2 to 8.2 microg/ml. Sulpiride toxicokinetic parameters such as t max = about 3 h, t 1/2 = 24.02 h, k(el) = 0.029 h(-1) were also estimated. They point out that the absorption rate is similar and the elimination is prorogated in sulpiride acute poisoning compared to therapeutic doses.

  5. Opportunistic pathogens in roof-captured rainwater samples, determined using quantitative PCR.

    PubMed

    Ahmed, W; Brandes, H; Gyawali, P; Sidhu, J P S; Toze, S

    2014-04-15

    In this study, quantitative PCR (qPCR) was used for the detection of four opportunistic bacterial pathogens in water samples collected from 72 rainwater tanks in Southeast Queensland, Australia. Tank water samples were also tested for fecal indicator bacteria (Escherichia coli and Enterococcus spp.) using culture-based methods. Among the 72 tank water samples tested, 74% and 94% samples contained E. coli and Enterococcus spp., respectively, and the numbers of E. coli and Enterococcus spp. in tank water samples ranged from 0.3 to 3.7 log₁₀ colony forming units (CFU) per 100 mL of water. In all, 29%, 15%, 13%, and 6% of tank water samples contained Aeromonas hydrophila, Staphylococcus aureus, Pseudomonas aeruginosa and Legionella pneumophila, respectively. The genomic units (GU) of opportunistic pathogens in tank water samples ranged from 1.5 to 4.6 log₁₀ GU per 100 mL of water. A significant correlation was found between E. coli and Enterococcus spp. numbers in pooled tank water samples data (Spearman's rs = 0.50; P < 0.001). In contrast, fecal indicator bacteria numbers did not correlate with the presence/absence of opportunistic pathogens tested in this study. Based on the results of this study, it would be prudent, to undertake a Quantitative Microbial Risk Assessment (QMRA) analysis of opportunistic pathogens to determine associated health risks for potable and nonpotable uses of tank water.

  6. Accurate determination of ⁴¹Ca concentrations in spent resins from the nuclear industry by accelerator mass spectrometry.

    PubMed

    Nottoli, Emmanuelle; Bourlès, Didier; Bienvenu, Philippe; Labet, Alexandre; Arnold, Maurice; Bertaux, Maité

    2013-12-01

    The radiological characterisation of nuclear waste is essential for managing storage sites. Determining the concentration of Long-Lived RadioNuclides (LLRN) is fundamental for their long-term management. This paper focuses on the measurement of low (41)Ca concentrations in ions exchange resins used for primary fluid purification in Pressurised Water Reactors (PWR). (41)Ca concentrations were successfully measured by Accelerator Mass Spectrometry (AMS) after the acid digestion of resin samples, followed by radioactive decontamination and isobaric suppression through successive hydroxide, carbonate, nitrate and final CaF2 precipitations. Measured (41)Ca concentrations ranged from 0.02 to 0.03 ng/g, i.e. from 0.06 to 0.09 Bq/g. The (41)Ca/(60)Co activity ratios obtained were remarkably reproducible and in good agreement with the current ratio used for resins management. PMID:24144617

  7. A quantitative thin layer chromatography method for determination of theophylline in plasma.

    PubMed

    Mirfazaelian, A; Goudarzi, M; Tabatabaiefar, M; Mahmoudian, M

    2002-01-01

    A simple assay method for theophylline in plasma using thin layer chromatography (TLC) was developed. The method involves extraction of the drug and internal standard (acetaminophen) by chloroform-isopropanol (75:25) followed by separation on TLC silica plates using a mixture of acetic acid, isopropanol, toluene (1: 12: 6), as the eluting solvent. Both peak height ratios and peak are ratios showed high correlation coefficient (r>0.98, p<0.001). However we used peak heights for the determinations. Within-day and between-day coefficients of variation were less than 4.4% and 7.8% respectively. The assay proved inexpensive, accurate and reproducible with a limit of detection of 100 ng/ml that makes it suitable for bioavailability studies.

  8. A method to accurately quantitate intensities of (32)P-DNA bands when multiple bands appear in a single lane of a gel is used to study dNTP insertion opposite a benzo[a]pyrene-dG adduct by Sulfolobus DNA polymerases Dpo4 and Dbh.

    PubMed

    Sholder, Gabriel; Loechler, Edward L

    2015-01-01

    Quantitating relative (32)P-band intensity in gels is desired, e.g., to study primer-extension kinetics of DNA polymerases (DNAPs). Following imaging, multiple (32)P-bands are often present in lanes. Though individual bands appear by eye to be simple and well-resolved, scanning reveals they are actually skewed-Gaussian in shape and neighboring bands are overlapping, which complicates quantitation, because slower migrating bands often have considerable contributions from the trailing edges of faster migrating bands. A method is described to accurately quantitate adjacent (32)P-bands, which relies on having a standard: a simple skewed-Gaussian curve from an analogous pure, single-component band (e.g., primer alone). This single-component scan/curve is superimposed on its corresponding band in an experimentally determined scan/curve containing multiple bands (e.g., generated in a primer-extension reaction); intensity exceeding the single-component scan/curve is attributed to other components (e.g., insertion products). Relative areas/intensities are determined via pixel analysis, from which relative molarity of components is computed. Common software is used. Commonly used alternative methods (e.g., drawing boxes around bands) are shown to be less accurate. Our method was used to study kinetics of dNTP primer-extension opposite a benzo[a]pyrene-N(2)-dG-adduct with four DNAPs, including Sulfolobus solfataricus Dpo4 and Sulfolobus acidocaldarius Dbh. Vmax/Km is similar for correct dCTP insertion with Dpo4 and Dbh. Compared to Dpo4, Dbh misinsertion is slower for dATP (∼20-fold), dGTP (∼110-fold) and dTTP (∼6-fold), due to decreases in Vmax. These findings provide support that Dbh is in the same Y-Family DNAP class as eukaryotic DNAP κ and bacterial DNAP IV, which accurately bypass N(2)-dG adducts, as well as establish the scan-method described herein as an accurate method to quantitate relative intensity of overlapping bands in a single lane, whether generated

  9. A method to accurately quantitate intensities of (32)P-DNA bands when multiple bands appear in a single lane of a gel is used to study dNTP insertion opposite a benzo[a]pyrene-dG adduct by Sulfolobus DNA polymerases Dpo4 and Dbh.

    PubMed

    Sholder, Gabriel; Loechler, Edward L

    2015-01-01

    Quantitating relative (32)P-band intensity in gels is desired, e.g., to study primer-extension kinetics of DNA polymerases (DNAPs). Following imaging, multiple (32)P-bands are often present in lanes. Though individual bands appear by eye to be simple and well-resolved, scanning reveals they are actually skewed-Gaussian in shape and neighboring bands are overlapping, which complicates quantitation, because slower migrating bands often have considerable contributions from the trailing edges of faster migrating bands. A method is described to accurately quantitate adjacent (32)P-bands, which relies on having a standard: a simple skewed-Gaussian curve from an analogous pure, single-component band (e.g., primer alone). This single-component scan/curve is superimposed on its corresponding band in an experimentally determined scan/curve containing multiple bands (e.g., generated in a primer-extension reaction); intensity exceeding the single-component scan/curve is attributed to other components (e.g., insertion products). Relative areas/intensities are determined via pixel analysis, from which relative molarity of components is computed. Common software is used. Commonly used alternative methods (e.g., drawing boxes around bands) are shown to be less accurate. Our method was used to study kinetics of dNTP primer-extension opposite a benzo[a]pyrene-N(2)-dG-adduct with four DNAPs, including Sulfolobus solfataricus Dpo4 and Sulfolobus acidocaldarius Dbh. Vmax/Km is similar for correct dCTP insertion with Dpo4 and Dbh. Compared to Dpo4, Dbh misinsertion is slower for dATP (∼20-fold), dGTP (∼110-fold) and dTTP (∼6-fold), due to decreases in Vmax. These findings provide support that Dbh is in the same Y-Family DNAP class as eukaryotic DNAP κ and bacterial DNAP IV, which accurately bypass N(2)-dG adducts, as well as establish the scan-method described herein as an accurate method to quantitate relative intensity of overlapping bands in a single lane, whether generated

  10. Simple saponification method for the quantitative determination of carotenoids in green vegetables.

    PubMed

    Larsen, Erik; Christensen, Lars P

    2005-08-24

    A simple, reliable, and gentle saponification method for the quantitative determination of carotenoids in green vegetables was developed. The method involves an extraction procedure with acetone and the selective removal of the chlorophylls and esterified fatty acids from the organic phase using a strongly basic resin (Ambersep 900 OH). Extracts from common green vegetables (beans, broccoli, green bell pepper, chive, lettuce, parsley, peas, and spinach) were analyzed by high-performance liquid chromatography (HPLC) for their content of major carotenoids before and after action of Ambersep 900 OH. The mean recovery percentages for most carotenoids [(all-E)-violaxanthin, (all-E)-lutein epoxide, (all-E)-lutein, neolutein A, and (all-E)-beta-carotene] after saponification of the vegetable extracts with Ambersep 900 OH were close to 100% (99-104%), while the mean recovery percentages of (9'Z)-neoxanthin increased to 119% and that of (all-E)-neoxanthin and neolutein B decreased to 90% and 72%, respectively.

  11. The renal quantitative scintillation camera study for determination of renal function

    SciTech Connect

    Thompson, I.M. Jr.; Boineau, F.G.; Evans, B.B.; Schlegel, J.U.

    1983-03-01

    The renal quantitative scintillation camera study assesses glomerular filtration rate and effective renal plasma flow based upon renal uptake of 99mtechnetium-iron ascorbate and 131iodine-hippuran, respectively. The method was compared to inulin, para-aminohippuric acid and creatinine clearance studies in 7 normal subjects and 9 patients with various degrees of reduced renal function. The reproducibility of the technique was determined in 15 randomly selected pediatric patients. The values of glomerular filtration rate and effective renal plasma flow were not significantly different from those of inulin and para-aminohippuric acid studies. The reproducibility of the technique was comparable to that of inulin and para-aminohippuric acid studies. Patient acceptance of the technique is excellent and the cost is minimal. Renal morphology and excretory dynamics also are demonstrated. The technique is advocated as a clinical measure of renal function.

  12. Ethanol determination in frozen fruit pulps: an application of quantitative nuclear magnetic resonance.

    PubMed

    da Silva Nunes, Wilian; de Oliveira, Caroline Silva; Alcantara, Glaucia Braz

    2016-04-01

    This study reports the chemical composition of five types of industrial frozen fruit pulps (acerola, cashew, grape, passion fruit and pineapple fruit pulps) and compares them with homemade pulps at two different stages of ripening. The fruit pulps were characterized by analyzing their metabolic profiles and determining their ethanol content using quantitative Nuclear Magnetic Resonance (qNMR). In addition, principal component analysis (PCA) was applied to extract more information from the NMR data. We detected ethanol in all industrial and homemade pulps; and acetic acid in cashew, grape and passion fruit industrial and homemade pulps. The ethanol content in some industrial pulps is above the level recommended by regulatory agencies and is near the levels of some post-ripened homemade pulps. This study demonstrates that qNMR can be used to rapidly detect ethanol content in frozen fruit pulps and food derivatives. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26578064

  13. Quantitative determination of mineral types and abundances from reflectance spectra using principal components analysis

    NASA Technical Reports Server (NTRS)

    Smith, M. O.; Adams, J. B.; Johnson, P. E.

    1985-01-01

    A procedure was developed for analyzing remote reflectance spectra, including multispectral images, that quantifies parameters such as types of mineral mixtures, the abundances of mixed minerals, and particle sizes. Principal components analysis reduced the spectral dimensionality and allowed testing the uniqueness and validity of spectral mixing models. By analyzing variations in the overall spectral reflectance curves, the type of spectral mixture was identified, mineral abundances quantified and the effects of particle size identified. The results demonstrate an advantage in classification accuracy over classical forms of analysis that ignore effects of particle-size or mineral-mixture systematics on spectra. The approach is applicable to remote sensing data of planetary surfaces for quantitative determinations of mineral abundances.

  14. ANSI/ASHRAE/IES Standard 90.1-2013 Determination of Energy Savings: Quantitative Analysis

    SciTech Connect

    Halverson, Mark A.; Athalye, Rahul A.; Rosenberg, Michael I.; Xie, YuLong; Wang, Weimin; Hart, Philip R.; Zhang, Jian; Goel, Supriya; Mendon, Vrushali V.

    2014-09-04

    This report provides a final quantitative analysis to assess whether buildings constructed according to the requirements of ANSI/ASHRAE/IES Standard 90.1-2013 would result in improved energy efficiency in commercial buildings. The final analysis considered each of the 110 addenda to Standard 90.1-2010 that were included in Standard 90.1-2013. PNNL reviewed all addenda included by ASHRAE in creating Standard 90.1-2013 from Standard 90.1-2010, and considered their combined impact on a suite of prototype building models across all U.S. climate zones. Most addenda were deemed to have little quantifiable impact on building efficiency for the purpose of DOE’s final determination. However, out of the 110 total addenda, 30 were identified as having a measureable and quantifiable impact.

  15. Characterization and quantitative determination of impurities in piperaquine phosphate by HPLC and LC/MS/MS.

    PubMed

    Dongre, Vaijanath G; Karmuse, Pravin P; Ghugare, Pradeep D; Gupta, Mukesh; Nerurkar, Bipin; Shaha, Chirag; Kumar, Ashok

    2007-01-01

    Four impurities in piperaquine phosphate bulk drug substance were detected by a newly developed gradient reverse phase high performance liquid chromatographic (HPLC) method. These impurities were identified by LC/MS/MS. The structures of impurities were confirmed by spectroscopic studies (NMR and IR) conducted using synthesized authentic compounds. The synthesized reference samples of the impurity compounds were used for the quantitative HPLC determination. The system suitability of HPLC analysis established the validity of the separation. The method was validated according to ICH guidelines with respect to specificity, precision, accuracy and linearity. Forced degradation studies were also performed for piperaquine phosphate bulk drug samples to demonstrate the stability indicating power of the newly developed HPLC method. PMID:16916594

  16. Quantitative determination of tilmicosin in canine serum by high performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Herrera, Michael; Ding, Haiqing; McClanahan, Robert; Owens, Jane G; Hunter, Robert P

    2007-09-15

    A highly sensitive and quantitative LC/MS/MS assay for the determination of tilmicosin in serum has been developed and validated. For sample preparation, 0.2 mL of canine serum was extracted with 3 mL of methyl tert-butyl ether. The organic layer was transferred to a new vessel and dried under nitrogen. The sample was then reconstituted for analysis by high performance liquid chromatography-tandem mass spectrometry. A Phenomenex Luna C8(2) analytical column was used for the chromatographic separation. The eluent was subsequently introduced to the mass spectrometer by electrospray ionization. A single range was validated for 50-5000 ng/mL for support of toxicokinetic studies. The inter-day relative error (inaccuracy) for the LLOQ samples ranged from -5.5% to 0.3%. The inter-day relative standard deviations (imprecision) at the respective LLOQ levels were < or =10.1%.

  17. Polymorphs of Aspirin Solid-state IR-LD spectroscopic and quantitative determination in solid mixtures

    NASA Astrophysics Data System (ADS)

    Koleva, Bojidarka B.

    2006-12-01

    Solid-state linear-dichroic infrared (IR-LD) spectroscopy, using an orientation technique as a suspension in nematic liquid crystal, has been carried out of Aspirin polymorphs (forms I and II). Reducing-difference procedure for polarized IR-spectra interpretation has been applied for structural analysis of both modifications and the data have been compared with known crystallographic ones. A vibration assignment of forms I and II has been included and on this basis, a quantitative determination by FT-IR spectra for form I in mixtures with second one has been presented, using intensity ratio of 1606 cm -1 peak (characteristic for both forms) to 599 cm -1 one (attributed to form I). The obtained reliability is 99.78%.

  18. Spectroscopic characterization and quantitative determination of atorvastatin calcium impurities by novel HPLC method

    NASA Astrophysics Data System (ADS)

    Gupta, Lokesh Kumar

    2012-11-01

    Seven process related impurities were identified by LC-MS in the atorvastatin calcium drug substance. These impurities were identified by LC-MS. The structure of impurities was confirmed by modern spectroscopic techniques like 1H NMR and IR and physicochemical studies conducted by using synthesized authentic reference compounds. The synthesized reference samples of the impurity compounds were used for the quantitative HPLC determination. These impurities were detected by newly developed gradient, reverse phase high performance liquid chromatographic (HPLC) method. The system suitability of HPLC analysis established the validity of the separation. The analytical method was validated according to International Conference of Harmonization (ICH) with respect to specificity, precision, accuracy, linearity, robustness and stability of analytical solutions to demonstrate the power of newly developed HPLC method.

  19. Hyaluronidase in ram semen. Quantitative determination, and isolation of multiple forms.

    PubMed Central

    Harrison, R A

    1988-01-01

    A study was made of hyaluronidase in ram semen. The end-group assay conditions used to determine activity quantitatively were chosen to ensure reliability as well as sensitivity [Gacesa, Savitsky, Dodgson & Olavesen (1981) Anal. Biochem. 118, 76-84]; they led to 1 W.H.O. Standard International Hyaluronidase Unit displaying 0.1263 EC munit (1 EC unit of activity releases 1 mumol equivalent of N-acetylglucosamine end groups/min at 37 degrees C). All the activity in the semen was shown to be sperm-derived, and intact spermatozoa were estimated to contain 1.23 EC units per 10(9) cells. In a low-ionic-strength medium, only some 20% of the hyaluronidase was extractable, although up to 80% of the activity could be extracted as the ionic strength was increased; further addition of detergent extracted the remainder. During purification of the enzyme, it was found that inclusion of poly(vinyl alcohol) in the media stabilized the activity; detergent inclusion also improved the yield, especially during early stages. As a consequence both of reliable quantitative determination and of stabilization, a number of forms of hyaluronidase could be isolated in high yield, by using anion-exchange chromatography, cation-exchange chromatography, affinity chromatography and gel filtration. The existence of all these forms was confirmed by electrophoresis and immunoblotting with the use of a monoclonal anti-(ram hyaluronidase) antibody, and their presence in very freshly prepared sperm extracts was demonstrated. The specific activity of the isolated major hyaluronidase form was 15.0 EC units/mg; this was equivalent to 119,000 W.H.O. units/mg, higher than any other previously reported values. Images Fig. 4. Fig. 5. PMID:3421926

  20. Effect of heterogeneity on the quantitative determination of trace elements in concrete.

    PubMed

    Weritz, Friederike; Schaurich, Dieter; Taffe, Alexander; Wilsch, Gerd

    2006-05-01

    Laser-induced breakdown spectroscopy has been used for quantitative measurement of trace elements, e.g. sulfur and chlorine, in concrete. Chloride and sulfate ions have a large effect on the durability of concrete structures, and quantitative measurement is important for condition assessment and quality assurance. Concrete is a highly heterogeneous material in composition and grain-size distribution, i.e. the spatial distribution of elements. Calibration plots were determined by use of laboratory-made reference samples consisting of pressings of cement powder, hydrated cement, cement mortar, and concrete, in which the heterogeneity of the material is increasing because of the aggregates. Coarse aggregate and cement paste are distinguishable by the intensity of the Ca spectral lines. More advanced evaluation is necessary to account for the effect of the fine aggregate. The three series of reference samples enable systematic study of the effects of heterogeneity on spectral intensity, signal fluctuation, uncertainty, and limits of detection. Spatially resolved measurements and many spectra enable statistical evaluation of the data. The heterogeneity has an effect on measurement of the sulfur and chlorine content, because both occur mainly in the cement matrix. Critical chloride concentrations are approximately 0.04% (m/m). The chlorine spectral line at 837.6 nm is evaluated. The natural sulfur content of concrete is approximately 0.1% (m/m). The spectral line at 921.3 nm is evaluated. One future application may be simultaneous determination of the amount of damaging trace elements and the cement content of the concrete. PMID:16520935

  1. Effect of heterogeneity on the quantitative determination of trace elements in concrete.

    PubMed

    Weritz, Friederike; Schaurich, Dieter; Taffe, Alexander; Wilsch, Gerd

    2006-05-01

    Laser-induced breakdown spectroscopy has been used for quantitative measurement of trace elements, e.g. sulfur and chlorine, in concrete. Chloride and sulfate ions have a large effect on the durability of concrete structures, and quantitative measurement is important for condition assessment and quality assurance. Concrete is a highly heterogeneous material in composition and grain-size distribution, i.e. the spatial distribution of elements. Calibration plots were determined by use of laboratory-made reference samples consisting of pressings of cement powder, hydrated cement, cement mortar, and concrete, in which the heterogeneity of the material is increasing because of the aggregates. Coarse aggregate and cement paste are distinguishable by the intensity of the Ca spectral lines. More advanced evaluation is necessary to account for the effect of the fine aggregate. The three series of reference samples enable systematic study of the effects of heterogeneity on spectral intensity, signal fluctuation, uncertainty, and limits of detection. Spatially resolved measurements and many spectra enable statistical evaluation of the data. The heterogeneity has an effect on measurement of the sulfur and chlorine content, because both occur mainly in the cement matrix. Critical chloride concentrations are approximately 0.04% (m/m). The chlorine spectral line at 837.6 nm is evaluated. The natural sulfur content of concrete is approximately 0.1% (m/m). The spectral line at 921.3 nm is evaluated. One future application may be simultaneous determination of the amount of damaging trace elements and the cement content of the concrete.

  2. Quantitative determination of sulfur content in concrete with laser-induced breakdown spectroscopy

    NASA Astrophysics Data System (ADS)

    Weritz, F.; Ryahi, S.; Schaurich, D.; Taffe, A.; Wilsch, G.

    2005-08-01

    Laser-induced breakdown spectroscopy has been employed for the investigation of the sulfur content of concrete. Sulfur compounds are a natural but minor component in building materials. The ingress of sulfates or sulfuric acid constitutes a major risk of chemical aggression for concrete. There is a need for a fast method, which can be used on-site and is able to investigate a wide range of different measuring points, so that damages can be characterized. For quantitative determination the sulfur spectral line at 921.3 nm is used. The optimum ambient atmosphere has been determined by comparison of measurements accomplished under air, argon and helium atmosphere. Reference samples have been produced and calibration curves have been determined, the results of LIBS measurements are compared with results from chemical analysis. Defining a limit for the intensity ratio of a calcium and a oxygen spectral line can reduce the influence of the heterogeneity of the material, so that only spectra with a high amount of cementitious material are evaluated. Depth profiles and spatial resolved sulfur distributions are presented measured on concrete cores originating from a highly sulfate contaminated clarifier.

  3. Simultaneous quantitative determinations of fluticasone propionate and salmeterol xinafote in diskus inhalers.

    PubMed

    Duran, A; Dogan, H N; Ulgen, M

    2014-01-01

    In the present study, two new methods were developed for the quantitative determination of active components of Seretide(®), commercially available pharmaceutical preparation in the diskus form. One of these methods was based on derivative spectrophotometry and used a zero-crossing technique. The determinations of fluticasone propionate and salmeterol xinafoate were performed by first order derivatisation at 216.5 nm and second order derivatisation at 250 nm, respectively. The concentration ranges were 5.0-32.5 μg/mL for fluticasone propionate and 2-12 μg/mL for salmeterol xinafoate. The second method developed also included high performance liquid chromatography. In this method, a methanol-water mobile phase mixture (95:5, v/v) and a C18 chromasil column as a stationary phase were used. The wavelength of the diode array UV detector was 260 nm; the flow rate was 1 mL/min. The concentration ranges were 2-16 μg/mL for fluticasone propionate and 1-8 μg/mL for salmeterol xinafoate. The results for both methods from diskus are in the pharmacopea limits. For the statistical determination of these results, these two methods were compared with t-test for the means and with F-test for the standard deviations. PMID:25313021

  4. Development of a rapid method for the quantitative determination of deoxynivalenol using Quenchbody.

    PubMed

    Yoshinari, Tomoya; Ohashi, Hiroyuki; Abe, Ryoji; Kaigome, Rena; Ohkawa, Hideo; Sugita-Konishi, Yoshiko

    2015-08-12

    Quenchbody (Q-body) is a novel fluorescent biosensor based on the antigen-dependent removal of a quenching effect on a fluorophore attached to antibody domains. In order to develop a method using Q-body for the quantitative determination of deoxynivalenol (DON), a trichothecene mycotoxin produced by some Fusarium species, anti-DON Q-body was synthesized from the sequence information of a monoclonal antibody specific to DON. When the purified anti-DON Q-body was mixed with DON, a dose-dependent increase in the fluorescence intensity was observed and the detection range was between 0.0003 and 3 mg L(-1). The coefficients of variation were 7.9% at 0.003 mg L(-1), 5.0% at 0.03 mg L(-1) and 13.7% at 0.3 mg L(-1), respectively. The limit of detection was 0.006 mg L(-1) for DON in wheat. The Q-body showed an antigen-dependent fluorescence enhancement even in the presence of wheat extracts. To validate the analytical method using Q-body, a spike-and-recovery experiment was performed using four spiked wheat samples. The recoveries were in the range of 94.9-100.2%. The concentrations of DON in twenty-one naturally contaminated wheat samples were quantitated by the Q-body method, LC-MS/MS and an immunochromatographic assay kit. The LC-MS/MS analysis showed that the levels of DON contamination in the samples were between 0.001 and 2.68 mg kg(-1). The concentrations of DON quantitated by LC-MS/MS were more strongly correlated with those using the Q-body method (R(2) = 0.9760) than the immunochromatographic assay kit (R(2) = 0.8824). These data indicate that the Q-body system for the determination of DON in wheat samples was successfully developed and Q-body is expected to have a range of applications in the field of food safety.

  5. Accurate determination of ochratoxin A in Korean fermented soybean paste by isotope dilution-liquid chromatography tandem mass spectrometry.

    PubMed

    Ahn, Seonghee; Lee, Suyoung; Lee, Joonhee; Kim, Byungjoo

    2016-01-01

    Ochratoxin A (OTA), a naturally occurring mycotoxin, has been frequently detected in doenjang, a traditional fermented soybean paste, when it is fermented under improper conditions. Reliable screening of OTA in traditional fermented soybean paste (doenjang) is a special food-safety issue in Korea. Our laboratory, the National Metrology Institute of Korea, established an isotope dilution-liquid chromatography tandem mass spectrometry (ID-LC/MS/MS) method as a higher-order reference method to be used for SI-traceable value-assignment of OTA in certified reference materials (CRMs). (13)C20-OTA was used as an internal standard. Sample preparation conditions and LC/MS measurement parameters were optimised for this purpose. The analytical method was validated by measuring samples fortified with OTA at various levels. Repeatability and reproducibility studies showed that the ID-LC/MS/MS method is reliable and reproducible within 2% relative standard deviation. The analytical method was applied to determine OTA in various commercial doenjang products and home-made doenjang products.

  6. A validated high-throughput UHPLC-MS/MS assay for accurate determination of rivaroxaban in plasma sample.

    PubMed

    Iqbal, Muzaffar; Khalil, Nasr Y; Imam, Faisal; Khalid Anwer, Md

    2015-01-01

    Rivaroxaban is a novel, selective and potent oral direct factor Xa inhibitor, therapeutically indicated in the treatment of thromboembolic diseases. Like traditional anticoagulants, routine coagulation monitoring of rivaroxaban is not necessary, but important in some clinical circumstances. In this study, a sensitive UHPLC-MS/MS assay for rapid determination of rivaroxaban in human plasma was developed and validated. Rivaroxaban and its internal standard (IS) were extracted from plasma using acetonitrile as protein precipitating agent. An isocratic mobile phase of acetonitrile: 10 mM ammonium acetate (80:20, v/v) at a flow rate of 0.3 mL/min was used for the separation of rivaroxaban and IS. Both rivaroxaban and IS was eluted within 1 min with a total run time of 1.5 min only. Electrospray ionization source in positive mode was used for the detections of rivaroxaban and IS. Precursor to product ion transition of m/z 436.00 > 144.87 for rivaroxaban and m/z 411.18 > 191.07 for IS were used in multiple reaction monitoring mode. Developed assay was fully validated in terms of selectivity, linearity, accuracy, precision, recovery, matrix effects and stability using official guideline on bioanalytical method. PMID:25106734

  7. The importance of suppressing spin diffusion effects in the accurate determination of the spatial structure of a flexible molecule by nuclear Overhauser effect spectroscopy

    NASA Astrophysics Data System (ADS)

    Khodov, I. A.; Efimov, S. V.; Klochkov, V. V.; Batista de Carvalho, L. A. E.; Kiselev, M. G.

    2016-02-01

    Two-dimensional nuclear Overhauser effect spectroscopy is applied to the elucidation of conformation distribution of small molecules in solution. An essential influence of the nonlinear multistep magnetization transfer (spin diffusion) on the NMR-based analysis of conformers distribution for small druglike molecules in solution was revealed. Therefore, the spin diffusion should be eliminated from the obtained NMR data in order to obtain accurate results. Efficiency of QUIET-NOESY spectroscopy in solving the problem of accurate determination of inter-proton distances in a small molecule was shown in a study of ibuprofen. Although it requires much experimental time, this technique was found to be helpful to solve the spin diffusion problem.

  8. Accurate determination of interface trap state parameters by admittance spectroscopy in the presence of a Schottky barrier contact: Application to ZnO-based solar cells

    NASA Astrophysics Data System (ADS)

    Marin, Andrew T.; Musselman, Kevin P.; MacManus-Driscoll, Judith L.

    2013-04-01

    This work shows that when a Schottky barrier is present in a photovoltaic device, such as in a device with an ITO/ZnO contact, equivalent circuit analysis must be performed with admittance spectroscopy to accurately determine the pn junction interface recombination parameters (i.e., capture cross section and density of trap states). Without equivalent circuit analysis, a Schottky barrier can produce an error of ˜4-orders of magnitude in the capture cross section and ˜50% error in the measured density of trap states. Using a solution processed ZnO/Cu2O photovoltaic test system, we apply our analysis to clearly separate the contributions of interface states at the pn junction from the Schottky barrier at the ITO/ZnO contact so that the interface state recombination parameters can be accurately characterized. This work is widely applicable to the multitude of photovoltaic devices, which use ZnO adjacent to ITO.

  9. Quantitative metallographic method for determining delta ferrite content in austenitic stainless steels. Final report

    SciTech Connect

    Pressly, G.A.

    1986-01-01

    Delta ferrite is a magnetic form of iron and has a body centered cubic crystal structure. It is often present as a nonequilibrium phase in austenitic stainless steel welds, castings, and wrought materials. The ferrite content of austenitic stainless steel can directly affect its properties, especially weldability and formability. Therefore, it is highly desirable to be able to predict and/or measure the ferrite content accurately. Current magnetic ferrite measuring methods are not applicable when test materials are geometrically small (less than 2.54 mm thick and 6.35 mm wide). Therefore, a standard metallographic test method STM 00107-A was established to determine delta ferrite content in small weldments and base metals of austenitic stainless steel. This standard test method (STM 00107-A) was then performed on several exemplary metallographic specimens to illustrate its capabilities and applications. The results from the exemplary tests were compared and contrasted to metallographic manual point count measurements, Ferritescope measurements, and predicted values calculated from chemical analyses. By utilizing the manual metallographic point count data, an accuracy of +-16% and a precision of +-0.77% were determined for the standard test method. The comparison of Ferritescope data to standard test method revealed that the results obtained by the two methods are close at low (0 to 3%) ferrite contents and Ferritscope results are substantially greater at higher (6 to 10%) ferrite contents. The standard test method data compiled from the exemplary weld specimens was noted to be very similar to the predicted values calculated from chemical analyses. It was also shown that because the standard test method utilizes optics the morphology of the delta ferrite particles can be determined. This type of determination is possible only with metallographic methods.

  10. GIS Integration for Quantitatively Determining the Capabilities of Five Remote Sensors for Resource Exploration

    NASA Technical Reports Server (NTRS)

    Pascucci, R. F.; Smith, A.

    1982-01-01

    To assist the U.S. Geological Survey in carrying out a Congressional mandate to investigate the use of side-looking airborne radar (SLAR) for resources exploration, a research program was conducted to define the contribution of SLAR imagery to structural geologic mapping and to compare this with contributions from other remote sensing systems. Imagery from two SLAR systems and from three other remote sensing systems was interpreted, and the resulting information was digitized, quantified and intercompared using a computer-assisted geographic information system (GIS). The study area covers approximately 10,000 square miles within the Naval Petroleum Reserve, Alaska, and is situated between the foothills of the Brooks Range and the North Slope. The principal objectives were: (1) to establish quantitatively, the total information contribution of each of the five remote sensing systems to the mapping of structural geology; (2) to determine the amount of information detected in common when the sensors are used in combination; and (3) to determine the amount of unique, incremental information detected by each sensor when used in combination with others. The remote sensor imagery that was investigated included real-aperture and synthetic-aperture radar imagery, standard and digitally enhanced LANDSAT MSS imagery, and aerial photos.

  11. Spectrophotometric Method for Quantitative Determination of Cefixime in Bulk and Pharmaceutical Preparation Using Ferroin Complex

    NASA Astrophysics Data System (ADS)

    Naeem Khan, M.; Qayum, A.; Ur Rehman, U.; Gulab, H.; Idrees, M.

    2015-09-01

    A method was developed for the quantitative determination of cefixime in bulk and pharmaceutical preparations using ferroin complex. The method is based on the oxidation of the cefixime with Fe(III) in acidic medium. The formed Fe(II) reacts with 1,10-phenanthroline, and the ferroin complex is measured spectrophotometrically at 510 nm against reagent blank. Beer's law was obeyed in the concentration range 0.2-10 μg/ml with a good correlation of 0.993. The molar absorptivity was calculated and was found to be 1.375×105 L/mol × cm. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.030 and 0.101 μg/ml respectively. The proposed method has reproducibility with a relative standard deviation of 5.28% (n = 6). The developed method was validated statistically by performing a recoveries study and successfully applied for the determination of cefixime in bulk powder and pharmaceutical formulations without interferences from common excipients. Percent recoveries were found to range from 98.00 to 102.05% for the pure form and 97.83 to 102.50% for pharmaceutical preparations.

  12. ANSI/ASHRAE/IESNA Standard 90.1-2010 Preliminary Determination Quantitative Analysis

    SciTech Connect

    Halverson, Mark A.; Liu, Bing; Rosenberg, Michael I.

    2010-11-01

    The United States (U.S.) Department of Energy (DOE) conducted a preliminary quantitative analysis to assess whether buildings constructed according to the requirements of the American National Standards Institute (ANSI)/American Society of Heating, Refrigerating and Air-Conditioning Engineers (ASHRAE)/Illuminating Engineering Society of North America (IESNA) Standard 90.1-2010 (ASHRAE Standard 90.1-2010, Standard 90.1-2010, or 2010 edition) would result in energy savings compared with buildings constructed to ANSI/ASHRAE/IESNA Standard 90.1-2007(ASHRAE Standard 90.1-2007, Standard 90.1-2007, or 2007 edition). The preliminary analysis considered each of the 109 addenda to ASHRAE Standard 90.1-2007 that were included in ASHRAE Standard 90.1-2010. All 109 addenda processed by ASHRAE in the creation of Standard 90.1-2010 from Standard 90.1-2007 were reviewed by DOE, and their combined impact on a suite of 16 building prototype models in 15 ASHRAE climate zones was considered. Most addenda were deemed to have little quantifiable impact on building efficiency for the purpose of DOE’s preliminary determination. However, out of the 109 addenda, 34 were preliminarily determined to have measureable and quantifiable impact.

  13. A Miniaturized Technique for Assessing Protein Thermodynamics and Function Using Fast Determination of Quantitative Cysteine Reactivity

    PubMed Central

    Isom, Daniel G.; Marguet, Philippe R.; Oas, Terrence G.; Hellinga, Homme W.

    2010-01-01

    Protein thermodynamic stability is a fundamental physical characteristic that determines biological function. Furthermore, alteration of thermodynamic stability by macromolecular interactions or biochemical modifications is a powerful tool for assessing the relationship between protein structure, stability, and biological function. High-throughput approaches for quantifying protein stability are beginning to emerge that enable thermodynamic measurements on small amounts of material, in short periods of time, and using readily accessible instrumentation. Here we present such a method, fast quantitative cysteine reactivity (fQCR), which exploits the linkage between protein stability, sidechain protection by protein structure, and structural dynamics to characterize the thermodynamic and kinetic properties of proteins. In this approach, the reaction of a protected cysteine and thiol-reactive fluorogenic indicator is monitored over a gradient of temperatures after a short incubation time. These labeling data can be used to determine the midpoint of thermal unfolding, measure the temperature dependence of protein stability, quantify ligand-binding affinity, and, under certain conditions, estimate folding rate constants. Here, we demonstrate the fQCR method by characterizing these thermodynamic and kinetic properties for variants of Staphylococcal nuclease and E. coli ribose-binding protein engineered to contain single, protected cysteines. These straightforward, information-rich experiments are likely to find applications in protein engineering and functional genomics. PMID:21387407

  14. ANSI/ASHRAE/IESNA Standard 90.1-2007 Final Determination Quantitative Analysis

    SciTech Connect

    Halverson, Mark A.; Liu, Bing; Richman, Eric E.; Winiarski, David W.

    2011-05-01

    The United States (U.S.) Department of Energy (DOE) conducted a final quantitative analysis to assess whether buildings constructed according to the requirements of the American National Standards Institute (ANSI)/American Society of Heating, Refrigerating and Air-Conditioning Engineers (ASHRAE)/Illuminating Engineering Society of North America (IESNA) Standard 90.1-2007 would result in energy savings compared with buildings constructed to ANSI/ASHRAE/IESNA Standard 90.1-2004. The final analysis considered each of the 44 addenda to ANSI/ASHRAE/IESNA Standard 90.1-2004 that were included in ANSI/ASHRAE/IESNA Standard 90.1-2007. All 44 addenda processed by ASHRAE in the creation of Standard 90.1-2007 from Standard 90.1-2004 were reviewed by DOE, and their combined impact on a suite of 15 building prototype models in 15 ASHRAE climate zones was considered. Most addenda were deemed to have little quantifiable impact on building efficiency for the purpose of DOE’s final determination. However, out of the 44 addenda, 9 were preliminarily determined to have measureable and quantifiable impact.

  15. Quantitative trait loci determining autogeny and body size in the Asian tiger mosquito (Aedes albopictus)

    PubMed Central

    Mori, A; Romero-Severson, J; Black, WC; Severson, DW

    2014-01-01

    The majority of mosquito species require a blood meal to stimulate vitellogenesis and subsequent oviposition (anautogeny), but some autogenous individuals complete their first ovarian cycle without a blood meal. Autogeny may be facultative or obligatory. In this study, we selected for an autogenous strain in the Asian tiger mosquito Aedes albopictus and examined an F1 intercross population for quantitative trait loci (QTL) determining the autogeny trait as well as wing length as a proxy for body size. Using composite interval mapping, we identified four QTL for each trait and observed considerable overlap in genome positions between each QTL for autogeny (follicle size) and wing length. Most QTL were minor in magnitude, individually explaining <10% of the phenotypic variation. Alleles from the autogenous parent generally showed a dominance or overdominance effect on both phenotypes. Strong genetic and phenotypic correlations indicate that autogeny and wing length are determined by up to four clusters of tightly linked genes or the potential pleiotropic effects of single genes. Although females from the autogenous strain produced ~5-fold more eggs following a blood meal than via autogeny, we suggest that maintenance of alleles for autogeny in natural populations is likely due to balancing selection. Autogeny should be favored under conditions of limited host availability for blood feeding or increased defensive behavior by the host and adequate larval nutrition. Correlation between autogeny and body size may reflect an increased ability for larger females to accumulate sufficient nutrient reserves to support oogenesis without a requirement for a blood meal. PMID:18461085

  16. Quantitative Microplate-Based Growth Assay for Determination of Antifungal Susceptibility of Histoplasma capsulatum Yeasts

    PubMed Central

    Goughenour, Kristie D.; Balada-Llasat, Joan-Miquel

    2015-01-01

    Standardized methodologies for determining the antifungal susceptibility of fungal pathogens is central to the clinical management of invasive fungal disease. Yeast-form fungi can be tested using broth macrodilution and microdilution assays. Reference procedures exist for Candida species and Cryptococcus yeasts; however, no standardized methods have been developed for testing the antifungal susceptibility of yeast forms of the dimorphic systemic fungal pathogens. For the dimorphic fungal pathogen Histoplasma capsulatum, susceptibility to echinocandins differs for the yeast and the filamentous forms, which highlights the need to employ Histoplasma yeasts, not hyphae, in antifungal susceptibility tests. To address this, we developed and optimized methodology for the 96-well microtiter plate-based measurement of Histoplasma yeast growth in vitro. Using optical density, the assay is quantitative for fungal growth with a dynamic range greater than 30-fold. Concentration and assay reaction time parameters were also optimized for colorimetric (MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction) and fluorescent (resazurin reduction) indicators of fungal vitality. We employed this microtiter-based assay to determine the antifungal susceptibility patterns of multiple clinical isolates of Histoplasma representing different phylogenetic groups. This methodology fulfills a critical need for the ability to monitor the effectiveness of antifungals on Histoplasma yeasts, the morphological form present in mammalian hosts and, thus, the form most relevant to disease. PMID:26246483

  17. Efficient Qualitative and Quantitative Determination of Antigen-induced Immune Responses*

    PubMed Central

    2016-01-01

    To determine the effectiveness of immunization strategies used in therapeutic antibody or vaccine development, it is critical to assess the quality of immunization-induced polyclonal antibody responses. Here, we developed a workflow that uses sensitive methods to quantitatively and qualitatively assess immune responses against foreign antigens with regard to antibody binding affinity and epitope diversity. The application of such detailed assessments throughout an immunization campaign can significantly reduce the resources required to generate highly specific antibodies. Our workflow consists of the following two steps: 1) the use of surface plasmon resonance to quantify antigen-specific antibodies and evaluate their apparent binding affinities, and 2) the recovery of serum IgGs using an automated small scale purification system, followed by the determination of their epitope diversity using hydrogen deuterium exchange coupled with mass spectrometry. We showed that these methods were sensitive enough to detect antigen-specific IgGs in the nanogram/μl range and that they provided information for differentiating the antibody responses of the various immunized animals that could not be obtained by conventional methods. We also showed that this workflow can guide the selection of an animal that produces high affinity antibodies with a desired epitope coverage profile, resulting in the generation of potential therapeutic monoclonal antibody clones with desirable functional profiles. We postulate that this workflow will be an important tool in the development of effective vaccines to combat the highly sophisticated evasion mechanisms of pathogens. PMID:27288409

  18. Quantitative Determination of Lateral Mode Dispersion in Film Bulk Acoustic Resonators through Laser Acoustic Imaging

    SciTech Connect

    Ken Telschow; John D. Larson III

    2006-10-01

    Film Bulk Acoustic Resonators are useful for many signal processing applications. Detailed knowledge of their operation properties are needed to optimize their design for specific applications. The finite size of these resonators precludes their use in single acoustic modes; rather, multiple wave modes, such as, lateral wave modes are always excited concurrently. In order to determine the contributions of these modes, we have been using a newly developed full-field laser acoustic imaging approach to directly measure their amplitude and phase throughout the resonator. This paper describes new results comparing modeling of both elastic and piezoelectric effects in the active material with imaging measurement of all excited modes. Fourier transformation of the acoustic amplitude and phase displacement images provides a quantitative determination of excited mode amplitude and wavenumber at any frequency. Images combined at several frequencies form a direct visualization of lateral mode excitation and dispersion for the device under test allowing mode identification and comparison with predicted operational properties. Discussion and analysis are presented for modes near the first longitudinal thickness resonance (~900 MHz) in an AlN thin film resonator. Plate wave modeling, taking account of material crystalline orientation, elastic and piezoelectric properties and overlayer metallic films, will be discussed in relation to direct image measurements.

  19. Quantitative determination of micronization-induced changes in the solid state of lactose.

    PubMed

    Della Bella, A; Müller, M; Soldati, L; Elviri, L; Bettini, R

    2016-05-30

    Lactose, in particular α-lactose monohydrate, is the most used carrier for inhalation. Its surface and solid-state properties play a key role in determining Dry Powder Inhalers (DPIs) performance. Techniques such as X-Ray Powder Diffraction (XRPD) and Differential Scanning Calorimetry (DSC), which are commonly used for the characterization of lactose, are not always capable of explaining the solid-state changes induced by processing, such as micronization. In the present work, the evaluation of the effect of the micronization process on the solid-state properties of lactose was carried out by XRPD and DSC and a satisfactory, although not unequivocal, interpretation of the thermal behaviour of lactose was obtained. Thus, a new gravimetric method correlating in a quantitative manner the weight change in specific sections of the Dynamic Vapour Sorption (DVS) profile and the amount of different forms of α-lactose (hygroscopic anhydrous, stable anhydrous and amorphous) simultaneously present in a given sample was developed and validated. The method is very simple and provides acceptable accuracy in phase quantitation (LOD=1.6, 2.4 and 2.7%, LOQ=5.4, 8.0 and 8.9% for hygroscopic anhydrous, stable anhydrous and amorphous α-lactose, respectively). The application of this method to a sample of micronized lactose led to results in agreement with those obtained by DSC and evidenced that hygroscopic anhydrous α-lactose, rather than amorphous lactose, can be generated in the micronization process. The proposed method may find a more general application for the quantification of polymorphs of compounds different than lactose, provided that the various solid phases afford different weight variations in specific regions of the DVS profile. PMID:27090154

  20. Quantitative determination of rarity of freshwater fishes and implications for imperiled-species designations.

    PubMed

    Pritt, Jeremy J; Frimpong, Emmanuel A

    2010-10-01

    Conserving rare species and protecting biodiversity and ecosystem functioning depends on sound information on the nature of rarity. Rarity is multidimensional and has a variety of definitions, which presents the need for a quantitative classification scheme with which to categorize species as rare or common. We constructed such a classification for North American freshwater fishes to better describe rarity in fishes and provide researchers and managers with a tool to streamline conservation efforts. We used data on range extents, habitat specificities, and local population sizes of North American freshwater fishes and a variety of quantitative methods and statistical decision criteria, including quantile regression and a cost-function algorithm to determine thresholds for categorizing a species as rare or common. Species fell into eight groups that conform to an established framework for rarity. Fishes listed by the American Fisheries Society (AFS) as endangered, threatened, or vulnerable were most often rare because their local population sizes were low, ranges were small, and they had specific habitat needs, in that order, whereas unlisted species were most often considered common on the basis of these three factors. Species with large ranges generally had few specific habitat needs, whereas those with small ranges tended to have narrow habitat specificities. We identified 30 species not designated as imperiled by AFS that were rare along all dimensions of rarity and may warrant further study or protection, and we found three designated species that were common along all dimensions and may require a review of their imperilment status. Our approach could be applied to other taxa to aid conservation decisions and serve as a useful tool for future revisions of listings of fish species. PMID:20337684

  1. Formation resistivity measurements from within a cased well used to quantitatively determine the amount of oil and gas present

    DOEpatents

    Vail, III, William B.

    1997-01-01

    Methods to quantitatively determine the separate amounts of oil and gas in a geological formation adjacent to a cased well using measurements of formation resistivity are disclosed. The steps include obtaining resistivity measurements from within a cased well of a given formation, obtaining the porosity, obtaining the resistivity of formation water present, computing the combined amounts of oil and gas present using Archie's Equations, determining the relative amounts of oil and gas present from measurements within a cased well, and then quantitatively determining the separate amounts of oil and gas present in the formation.

  2. Formation resistivity measurements from within a cased well used to quantitatively determine the amount of oil and gas present

    DOEpatents

    Vail, W.B. III

    1997-05-27

    Methods to quantitatively determine the separate amounts of oil and gas in a geological formation adjacent to a cased well using measurements of formation resistivity are disclosed. The steps include obtaining resistivity measurements from within a cased well of a given formation, obtaining the porosity, obtaining the resistivity of formation water present, computing the combined amounts of oil and gas present using Archie`s Equations, determining the relative amounts of oil and gas present from measurements within a cased well, and then quantitatively determining the separate amounts of oil and gas present in the formation. 7 figs.

  3. Accurate determination of small one-bond heteronuclear residual dipolar couplings by F1 coupled HSQC modified with a G-BIRD (r) module

    NASA Astrophysics Data System (ADS)

    Fehér, Krisztina; Berger, Stefan; Kövér, Katalin E.

    2003-08-01

    We report a G-BIRD (r) modified coupled HSQC experiment for the accurate determination of one-bond heteronuclear residual dipolar couplings. The G-BIRD (r) module has been employed to refocus the long-range coupling evolution of the heteronucleus during the t1 frequency labeling period. As a result, the crosspeaks obtained are split by only the direct one-bond coupling that can be extracted by measuring simple frequency differences between singlet maxima. Additionally the decoupling of long-range multiple bond splittings leads to considerable sensitivity enhancement. The modification also has been applied in a TROSY sequence resulting in a significant sensitivity and resolution improvement.

  4. Accurate determination of the fine-structure intervals in the 3P ground states of C-13 and C-12 by far-infrared laser magnetic resonance

    NASA Technical Reports Server (NTRS)

    Cooksy, A. L.; Saykally, R. J.; Brown, J. M.; Evenson, K. M.

    1986-01-01

    Accurate values are presented for the fine-structure intervals in the 3P ground state of neutral atomic C-12 and C-13 as obtained from laser magnetic resonance spectroscopy. The rigorous analysis of C-13 hyperfine structure, the measurement of resonant fields for C-12 transitions at several additional far-infrared laser frequencies, and the increased precision of the C-12 measurements, permit significant improvement in the evaluation of these energies relative to earlier work. These results will expedite the direct and precise measurement of these transitions in interstellar sources and should assist in the determination of the interstellar C-12/C-13 abundance ratio.

  5. [Effect of algorithms for calibration set selection on quantitatively determining asiaticoside content in Centella total glucosides by near infrared spectroscopy].

    PubMed

    Zhan, Xue-yan; Zhao, Na; Lin, Zhao-zhou; Wu, Zhi-sheng; Yuan, Rui-juan; Qiao, Yan-jiang

    2014-12-01

    The appropriate algorithm for calibration set selection was one of the key technologies for a good NIR quantitative model. There are different algorithms for calibration set selection, such as Random Sampling (RS) algorithm, Conventional Selection (CS) algorithm, Kennard-Stone(KS) algorithm and Sample set Portioning based on joint x-y distance (SPXY) algorithm, et al. However, there lack systematic comparisons between two algorithms of the above algorithms. The NIR quantitative models to determine the asiaticoside content in Centella total glucosides were established in the present paper, of which 7 indexes were classified and selected, and the effects of CS algorithm, KS algorithm and SPXY algorithm for calibration set selection on the accuracy and robustness of NIR quantitative models were investigated. The accuracy indexes of NIR quantitative models with calibration set selected by SPXY algorithm were significantly different from that with calibration set selected by CS algorithm or KS algorithm, while the robustness indexes, such as RMSECV and |RMSEP-RMSEC|, were not significantly different. Therefore, SPXY algorithm for calibration set selection could improve the predicative accuracy of NIR quantitative models to determine asiaticoside content in Centella total glucosides, and have no significant effect on the robustness of the models, which provides a reference to determine the appropriate algorithm for calibration set selection when NIR quantitative models are established for the solid system of traditional Chinese medcine.

  6. [Effect of algorithms for calibration set selection on quantitatively determining asiaticoside content in Centella total glucosides by near infrared spectroscopy].

    PubMed

    Zhan, Xue-yan; Zhao, Na; Lin, Zhao-zhou; Wu, Zhi-sheng; Yuan, Rui-juan; Qiao, Yan-jiang

    2014-12-01

    The appropriate algorithm for calibration set selection was one of the key technologies for a good NIR quantitative model. There are different algorithms for calibration set selection, such as Random Sampling (RS) algorithm, Conventional Selection (CS) algorithm, Kennard-Stone(KS) algorithm and Sample set Portioning based on joint x-y distance (SPXY) algorithm, et al. However, there lack systematic comparisons between two algorithms of the above algorithms. The NIR quantitative models to determine the asiaticoside content in Centella total glucosides were established in the present paper, of which 7 indexes were classified and selected, and the effects of CS algorithm, KS algorithm and SPXY algorithm for calibration set selection on the accuracy and robustness of NIR quantitative models were investigated. The accuracy indexes of NIR quantitative models with calibration set selected by SPXY algorithm were significantly different from that with calibration set selected by CS algorithm or KS algorithm, while the robustness indexes, such as RMSECV and |RMSEP-RMSEC|, were not significantly different. Therefore, SPXY algorithm for calibration set selection could improve the predicative accuracy of NIR quantitative models to determine asiaticoside content in Centella total glucosides, and have no significant effect on the robustness of the models, which provides a reference to determine the appropriate algorithm for calibration set selection when NIR quantitative models are established for the solid system of traditional Chinese medcine. PMID:25881421

  7. Development of a quantitative GC-FID method for the determination of sucrose mono- and diesters in foods.

    PubMed

    Cucu, Tatiana; De Meulenaer, Bruno

    2015-01-01

    Sucrose esters (E 473) are emulsifiers used in foods to improve different technological properties. They should conform to the specifications laid down in Commission Regulation No. 231/2012 and be used at amounts not exceeding the maximal ones set by Commission Regulation No. 1129/2011. In order to be able to characterise commercial sucrose ester formulations and to evaluate whether they are used correctly by the food industry, a quantitative GC-FID method was developed. Standards of monoesters and diesters were isolated from commercial additive preparations because no commercial ones were available. Commercial sucrose monolaureate and in-house-synthesised sucrose diarachidonate were used as internal standards. The method showed limits of detection and quantification of 2.9 and 5.7 µg ml(-1) respectively for the monoesters and 42.8 and 129.7 µg ml(-1) respectively for the diesters. The analysed commercial additive formulations contained mainly mono- and diesters of palmitic and stearic acid with low amounts of free fatty acid and sucrose. Different food matrices were incurred with commercial sucrose esters formulations and recoveries ranged between 92% and 118% for the monoesters and between 77% and 120% for the diesters. Recovery of sucrose monoesters in cake was around 34% when no enzymatic treatment was applied, and about 64% when enzymatic treatment with Clara-Diastase was applied. This indicated that sucrose esters can interact strongly with the matrix during food production and that treatment with enzymes is essential to determine the esters' content accurately in some classes of food products.

  8. Hepatotoxic potential of asarones: in vitro evaluation of hepatotoxicity and quantitative determination in herbal products

    PubMed Central

    Patel, Dhavalkumar N.; Ho, Han K.; Tan, Liesbet L.; Tan, Mui-Mui B.; Zhang, Qian; Low, Min-Yong; Chan, Cheng-Leng; Koh, Hwee-Ling

    2015-01-01

    α and β asarones are natural constituents of some aromatic plants, especially species of the genus Acorus (Araceae). In addition to reports of beneficial properties of asarones, genotoxicity and carcinogenicity are also reported. Due to potential toxic effects of β-asarone, a limit of exposure from herbal products of ~2 μg/kg body weight/day has been set temporarily until a full benefit/risk assessment has been carried out by the European Medicines Agency. Therefore, it is important to monitor levels of β-asarone in herbal products. In this study, we developed a simple, rapid and validated GC-MS method for quantitative determination of asarones and applied it in 20 pediatric herbal products after detecting high concentrations of β-asarone in a product suspected to be implicated in hepatotoxicity in a 3 month old infant. Furthermore, targeted toxicological effects were further investigated in human hepatocytes (THLE-2 cells) by employing various in vitro assays, with the goal of elucidating possible mechanisms for the observed toxicity. Results showed that some of the products contained as much as 4–25 times greater amounts of β-asarone than the recommended levels. In 4 of 10 samples found to contain asarones, the presence of asarones could not be linked to the labeled ingredients, possibly due to poor quality control. Cell-based investigations in THLE-2 cells confirmed the cytotoxicity of β-asarone (IC50 = 40.0 ± 2.0 μg/mL) which was associated with significant lipid peroxidation and glutathione depletion. This observed cytotoxic effect is likely due to induction of oxidative stress by asarones. Overall, the results of this study ascertained the usability of this GC-MS method for the quantitative determination of asarones from herbal products, and shed light on the importance of controlling the concentration of potentially toxic asarones in herbal products to safeguard consumer safety, especially when the target consumers are young children. Further

  9. Comparison of a quantitative microtiter method, a quantitative automated method, and the plate-count method for determining microbial complement resistance.

    PubMed

    Lee, M D; Wooley, R E; Brown, J; Spears, K R; Nolan, L K; Shotts, E B

    1991-01-01

    A quantitative microtiter method for determining the degree of complement resistance or sensitivity of microorganisms is described. The microtiter method is compared with a quantitative automated system and the standard plate-count technique. Data were accumulated from 30 avian Escherichia coli isolates incubated at 35 C with either chicken plasma or heat-inactivated chicken plasma. Analysis of data generated by the automated system and plate-count techniques resulted in a classification of the microorganisms into three groups: those sensitive to the action of complement; those of intermediate sensitivity to the action of complement; and those resistant to the action of complement. Although the three methods studied did not agree absolutely, there were statistically significant correlations among them.

  10. 13C-labeled gluconate tracing as a direct and accurate method for determining the pentose phosphate pathway split ratio in Penicillium chrysogenum.

    PubMed

    Kleijn, Roelco J; van Winden, Wouter A; Ras, Cor; van Gulik, Walter M; Schipper, Dick; Heijnen, Joseph J

    2006-07-01

    In this study we developed a new method for accurately determining the pentose phosphate pathway (PPP) split ratio, an important metabolic parameter in the primary metabolism of a cell. This method is based on simultaneous feeding of unlabeled glucose and trace amounts of [U-13C]gluconate, followed by measurement of the mass isotopomers of the intracellular metabolites surrounding the 6-phosphogluconate node. The gluconate tracer method was used with a penicillin G-producing chemostat culture of the filamentous fungus Penicillium chrysogenum. For comparison, a 13C-labeling-based metabolic flux analysis (MFA) was performed for glycolysis and the PPP of P. chrysogenum. For the first time mass isotopomer measurements of 13C-labeled primary metabolites are reported for P. chrysogenum and used for a 13C-based MFA. Estimation of the PPP split ratio of P. chrysogenum at a growth rate of 0.02 h(-1) yielded comparable values for the gluconate tracer method and the 13C-based MFA method, 51.8% and 51.1%, respectively. A sensitivity analysis of the estimated PPP split ratios showed that the 95% confidence interval was almost threefold smaller for the gluconate tracer method than for the 13C-based MFA method (40.0 to 63.5% and 46.0 to 56.5%, respectively). From these results we concluded that the gluconate tracer method permits accurate determination of the PPP split ratio but provides no information about the remaining cellular metabolism, while the 13C-based MFA method permits estimation of multiple fluxes but provides a less accurate estimate of the PPP split ratio.

  11. Quantitative modal determination of geological samples based on X-ray multielemental map acquisition.

    PubMed

    Cossio, Roberto; Borghi, Alessandro; Ruffini, Raffaella

    2002-04-01

    Multielemental X-ray maps collected by a remote scanning system of the electron beam are processed by a dedicated software program performing accurate modal determination of geological samples. The classification of different mineral phases is based on elemental concentrations. The software program Petromod loads the maps into a database and computes a matrix consisting of numerical values proportional to the elemental concentrations. After an initial calibration, the program can perform the chemical composition calculated on the basis of a fixed number of oxygens for a selected area. In this way, it is possible to identify all the mineral phases occurring in the sample. Up to three elements can be selected to calculate the modal percentage of the identified mineral. An automated routine scans the whole set of maps and assigns each pixel that satisfies the imposed requirements to the selected phase. Repeating this procedure for every mineral phase occurring in the mapped area, a modal distribution of the rock-forming minerals can be performed. The final output consists of a digitized image, which can be further analyzed by common image analysis software, and a table containing the calculated modal percentages. The method is here applied to a volcanic and a metamorphic rock sample. PMID:12533243

  12. Quantitative acylcarnitine determination by UHPLC-MS/MS--Going beyond tandem MS acylcarnitine "profiles".

    PubMed

    Minkler, Paul E; Stoll, Maria S K; Ingalls, Stephen T; Kerner, Janos; Hoppel, Charles L

    2015-12-01

    Tandem MS "profiling" of acylcarnitines and amino acids was conceived as a first-tier screening method, and its application to expanded newborn screening has been enormously successful. However, unlike amino acid screening (which uses amino acid analysis as its second-tier validation of screening results), acylcarnitine "profiling" also assumed the role of second-tier validation, due to the lack of a generally accepted second-tier acylcarnitine determination method. In this report, we present results from the application of our validated UHPLC-MS/MS second-tier method for the quantification of total carnitine, free carnitine, butyrobetaine, and acylcarnitines to patient samples with known diagnoses: malonic acidemia, short-chain acyl-CoA dehydrogenase deficiency (SCADD) or isobutyryl-CoA dehydrogenase deficiency (IBD), 3-methyl-crotonyl carboxylase deficiency (3-MCC) or ß-ketothiolase deficiency (BKT), and methylmalonic acidemia (MMA). We demonstrate the assay's ability to separate constitutional isomers and diastereomeric acylcarnitines and generate values with a high level of accuracy and precision. These capabilities are unavailable when using tandem MS "profiles". We also show examples of research interest, where separation of acylcarnitine species and accurate and precise acylcarnitine quantification is necessary.

  13. Quantitative modal determination of geological samples based on X-ray multielemental map acquisition.

    PubMed

    Cossio, Roberto; Borghi, Alessandro; Ruffini, Raffaella

    2002-04-01

    Multielemental X-ray maps collected by a remote scanning system of the electron beam are processed by a dedicated software program performing accurate modal determination of geological samples. The classification of different mineral phases is based on elemental concentrations. The software program Petromod loads the maps into a database and computes a matrix consisting of numerical values proportional to the elemental concentrations. After an initial calibration, the program can perform the chemical composition calculated on the basis of a fixed number of oxygens for a selected area. In this way, it is possible to identify all the mineral phases occurring in the sample. Up to three elements can be selected to calculate the modal percentage of the identified mineral. An automated routine scans the whole set of maps and assigns each pixel that satisfies the imposed requirements to the selected phase. Repeating this procedure for every mineral phase occurring in the mapped area, a modal distribution of the rock-forming minerals can be performed. The final output consists of a digitized image, which can be further analyzed by common image analysis software, and a table containing the calculated modal percentages. The method is here applied to a volcanic and a metamorphic rock sample.

  14. Quantitative determination of oxygen yield in a chemical oxygen-iodine laser

    NASA Astrophysics Data System (ADS)

    Kendrick, Kip R.; Helms, Charles A.; Quillen, Brian; Copland, R. J.

    1998-05-01

    With the advent of the Airborne Laser program, the emphasis of chemical oxygen-iodine laser (COIL) research has shifted toward improving the overall efficiency. A key component of COIL is the singlet-oxygen generator (SOG). To asses the efficiency of the SOG an accurate method of determining the yield of O2((alpha) 1(Delta) g),[O2((alpha) 1(Delta) g)]/[O2(total)] where [O2(total)]equals[O2((alpha) 1(Delta) g)]+[O2(X3(Sigma) g-)], has been developed. Absorption measurements of ground-state oxygen utilizing the magnetic-dipole transition, O2(X3(Sigma) g-) at 763 nm, have been obtained using a diode laser in conjunction with a multiple-pass Herriot-cell on a 10 kW class supersonic SOIL (RADICL). When RADICL is configured with a 0.35' throat, 15' diskpack, and a medium volume transition duct, with a diluent ratio (He:O2) of 3:1, the yield of O2((alpha) 1(Delta) g) in the diagnostic duct is 0.41 +/- 0.02.

  15. Quantitative and isomeric determination of amphetamine and methamphetamine from urine using a nonprotic elution solvent and R(-)-alpha-methoxy-alpha-trifluoromethylphenylacetic acid chloride derivatization.

    PubMed

    Holler, Justin M; Vorce, Shawn P; Bosy, Thomas Z; Jacobs, Aaron

    2005-10-01

    ). The method allows for a simple and accurate quantitation and isomeric determination of amphetamine and methamphetamine using a process that eliminates extraction and derivatization complications common in current methods.

  16. Quantitative gas chromatographic mass spectrometric determination of pinaverium-bromide in human serum.

    PubMed

    de Weerdt, G A; Beke, R P; Verdievel, H G; Barbier, F; Jonckheere, J A; de Leenheer, A P

    1983-03-01

    A method has been developed for quantitative determination of pinaverium-bromide, a quaternary ammonium derivative with papaverine-like activity, in human serum. The method involves a chloroform extraction of serum spiked with N-(6,6-dimethyl bicyclo[3.1.1]2-heptenyl-ethoxyethyl) perhydro-1,4-oxazine as internal standard. After evaporation of the solvent, and reduction of the residue with Raney-Nickel, the internal standard and the reduced pinaverium-bromide are re-extracted from the reaction mixture with toluene and analysed isothermally on a fused silica column coated with OV-101. Although chemical ionization with methane revealed intense protonated molecular ions for both pinaverium-bromide and the internal standard, selectivity and sensitivity were significantly lower in comparison with electron impact ionization at 70 e V. Therefore, quantification was performed in the electron impact mode by single ion monitoring of the common fragment ion at m/z 100.2. A linear detector response was observed up to 160 ng ml-1. A within-run assay precision better than 2% CV (n = 5) was found, and a detection limit of 1 ng pinaverium-bromide ml-1 of serum was attained. PMID:6850068

  17. Quantitative Determination of Catechin as Chemical Marker in Pediatric Polyherbal Syrup by HPLC/DAD.

    PubMed

    Sheikh, Zeeshan A; Siddiqui, Zafar A; Naveed, Safila; Usmanghani, Khan

    2016-09-01

    Vivabon syrup is a balanced composition of dietary ingredients of phytopharmaceutical nature for maintaining the physique, vigor, vitality and balanced growth of children. The herbal ingredients of pediatric syrup are rich in bioflavonoid, proteins, vitamins, glycosides and trace elements. Vivabon is formulated with herbal drugs such as Phoenix sylvestris, Emblica officinalis, Withania somnifera, Centella asiatica, Amomum subulatum, Zingiber officinalis, Trigonella foenum-graecum, Centaurea behen and Piper longum Catechins are flavan-3-ols that are found widely in the medicinal herbs and are utilized for anti-inflammatory, cardio protective, hepato-protective, neural protection and other biological activities. In general, the dietary intake of flavonoids has been regarded traditionally as beneficial for body growth. Standardization of Vivabon syrup dosage form using HPLC/DAD has been developed for quantitative estimation of Catechin as a chemical marker. The method was validated as per ICH guidelines. Validation studies demonstrated that the developed HPLC method is quite distinct, reproducible as well as quick and fast. The relatively high recovery and low comparable standard deviation confirm the suitability of the developed method for the determination of Catechin in syrup. PMID:27165575

  18. Determination of single photon ionization cross sections for quantitative analysis of complex organic mixtures.

    PubMed

    Adam, Thomas; Zimmermann, Ralf

    2007-11-01

    Soft single photon ionization (SPI)-time-of-flight mass spectrometry (TOFMS) is well suited for fast and comprehensive analysis of complex organic gas mixtures, which has been demonstrated in various applications. This work describes a calibration scheme for SPI, which enables quantification of a large number of compounds by only calibrating one compound of choice, in this case benzene. Photoionization cross sections of 22 substances were determined and related to the yield of benzene. These substances included six alkanes (pentane, hexane, heptane, octane, nonane, decane), three alkenes (propene, butane, pentene), two alkynes (propyne, butyne), two dienes (butadiene, isoprene), five monoaromatic species (benzene, toluene, xylene, styrene, monochlorobenzene) and NO. The cross sections of organic compounds differ by about one order of magnitude but the photoionization properties of compounds belonging to one compound class are rather similar. Therefore, the scheme can also be used for an approximate quantification of compound classes. This is demonstrated by a fast characterization and pattern recognition of two gasoline samples with different origins (Germany and South Africa) and a diesel sample (Germany). The on-line capability of the technique and the scheme is demonstrated by quantitatively monitoring and comparing the cold engine start of four vehicles: a gasoline passenger car, a diesel van, a motorbike and a two-stroke scooter.

  19. Sensitive time-resolved fluoroimmunoassay for quantitative determination of clothianidin in agricultural samples.

    PubMed

    Li, Ming; Sheng, Enze; Yuan, Yulong; Liu, Xiaofeng; Hua, Xiude; Wang, Minghua

    2014-05-01

    Europium (Eu(3+))-labeled antibody was used as a fluorescent label to develop a highly sensitive time-resolved fluoroimmunoassay (TRFIA) for determination of clothianidin residues in agricultural samples. Toward this goal, the Eu(3+)-labeled polyclonal antibody and goat anti-rabbit antibody were prepared for developing and evaluating direct competitive TRFIA (dc-TRFIA) and indirect competitive TRFIA (ic-TRFIA). Under optimal conditions, the half-maximal inhibition concentration (IC50) and the limit of detection (LOD, IC10) of clothianidin were 9.20 and 0.0909 μg/L for the dc-TRFIA and 2.07 and 0.0220 μg/L for the ic-TRFIA, respectively. The ic-TRFIA has no obvious cross-reactivity with the analogues of clothianidin except for dinotefuran. The average recoveries of clothianidin from spiked water, soil, cabbage, and rice samples were estimated to range from 74.1 to 115.9 %, with relative standard deviations of 3.3 to 11.7 %. The results of TRFIA for the blind samples were largely consistent with gas chromatography (R (2) = 0.9902). The optimized ic-TRFIA might become a sensitive and satisfactory analytical method for the quantitative monitoring of clothianidin residues in agricultural samples.

  20. Determination of internal controls for quantitative gene expression of Isochrysis zhangjiangensis at nitrogen stress condition

    NASA Astrophysics Data System (ADS)

    Wu, Shuang; Zhou, Jiannan; Cao, Xupeng; Xue, Song

    2016-02-01

    Isochrysis zhangjiangensis is a potential marine microalga for biodiesel production, which accumulates lipid under nitrogen limitation conditions, but the mechanism on molecular level is veiled. Quantitative real-time polymerase chain reaction (qPCR) provides the possibility to investigate the gene expression levels, and a valid reference for data normalization is an essential prerequisite for firing up the analysis. In this study, five housekeeping genes, actin (ACT), α-tubulin (TUA), ß-tubulin (TUB), ubiquitin (UBI), 18S rRNA (18S) and one target gene, diacylglycerol acyltransferase (DGAT), were used for determining the reference. By analyzing the stabilities based on calculation of the stability index and on operating the two types of software, geNorm and bestkeeper, it showed that the reference genes widely used in higher plant and microalgae, such as UBI, TUA and 18S, were not the most stable ones in nitrogen-stressed I. zhangjiangensis, and thus are not suitable for exploring the mRNA expression levels under these experimental conditions. Our results show that ACT together with TUB is the most feasible internal control for investigating gene expression under nitrogen-stressed conditions. Our findings will contribute not only to future qPCR studies of I. zhangjiangensis, but also to verification of comparative transcriptomics studies of the microalgae under similar conditions.

  1. An Evaluation of Quantitative Methods of Determining the Degree of Melting Experienced by a Chondrule

    NASA Technical Reports Server (NTRS)

    Nettles, J. W.; Lofgren, G. E.; Carlson, W. D.; McSween, H. Y., Jr.

    2004-01-01

    Many workers have considered the degree to which partial melting occurred in chondrules they have studied, and this has led to attempts to find reliable methods of determining the degree of melting. At least two quantitative methods have been used in the literature: a convolution index (CVI), which is a ratio of the perimeter of the chondrule as seen in thin section divided by the perimeter of a circle with the same area as the chondrule, and nominal grain size (NGS), which is the inverse square root of the number density of olivines and pyroxenes in a chondrule (again, as seen in thin section). We have evaluated both nominal grain size and convolution index as melting indicators. Nominal grain size was measured on the results of a set of dynamic crystallization experiments previously described, where aliquots of LEW97008(L3.4) were heated to peak temperatures of 1250, 1350, 1370, and 1450 C, representing varying degrees of partial melting of the starting material. Nominal grain size numbers should correlate with peak temperature (and therefore degree of partial melting) if it is a good melting indicator. The convolution index is not directly testable with these experiments because the experiments do not actually create chondrules (and therefore they have no outline on which to measure a CVI). Thus we had no means to directly test how well the CVI predicted different degrees of melting. Therefore, we discuss the use of the CVI measurement and support the discussion with X-ray Computed Tomography (CT) data.

  2. Quantitative determination of fluorotelomer sulfonates in groundwater by LC MS/MS.

    PubMed

    Schultz, Melissa M; Barofsky, Douglas F; Field, Jennifer A

    2004-03-15

    Aqueous film-forming foams (AFFF) are complex mixtures containing fluorocarbon- and hydrocarbon-based surfactants that are used to fight hydrocarbon-fueled fires. The military is the largest consumer of AFFF in the United States, and fire-training activities conducted at military bases have led to groundwater contamination by unspent fuels and AFFF chemicals. A direct-injection, liquid-chromatography tandem mass spectrometry (LC MS/MS) method was developed to quantify a suite of fluorotelomer sulfonate surfactants in groundwater collected from military bases where fire-training activities were conducted. The 4:2, 6:2, and 8:2 fluorotelomer sulfonates were detected and quantified in groundwater from two of the three military bases. The total fluorotelomer sulfonate concentrations observed at Wurtsmith AFB, MI, and Tyndall AFB, FL, ranged respectively from below quantitation (< or = 0.60) to 182 microg/L and from 1100 to 14,600 microg/L. Analyses of a fluorotelomer-based AFFF concentrate by negative ion fast atom bombardment/mass spectrometry and LC MS/MS analyses indicate that the AFFF concentrate contains only a small amount of fluorotelomer sulfonates and that fluoroalkylthioamido sulfonates are the main anionic fluorosurfactant in the mixtures. More research is needed to determine the fate of fluoroalkylthioamido sulfonates in the environment.

  3. Identification and quantitative determination of lignans in Cedrus atlantica resins using 13C NMR spectroscopy.

    PubMed

    Nam, Anne-Marie; Paoli, Mathieu; Castola, Vincent; Casanova, Joseph; Bighelli, Ange

    2011-03-01

    Identification and quantitative determination of individual components of resin collected on the trunk of 28 Cedrus atlantica trees, grown in Corsica, has been carried out using 13C NMR spectroscopy. Eight resin acids bearing either the pimarane or abietane skeleton, two monoterpene hydrocarbons and four oxygenated neutral diterpenes have been identified, as well as three lignans, scarcely found in resins. Three groups could be distinguished within the 28 resin samples. The nine samples of Group I had their composition dominated by diterpene acids (33.7-45.8%), with abietic acid (6.2-18.7%) and isopimaric acid (5.1-12.6%) being the major components. The four samples of Group II contained resin acids (main components) and lignans in moderate amounts (up to 10.3%). Conversely, lignans (38.8-63.8%) were by far the major components of the 15 samples of Group III. Depending on the sample, the major component was pinoresinol (18.1-38.9%), lariciresinol (17.2-33.7%) or lariciresinol 9'-acetate (16.9-29.1%). Finally, due to the high biological interest in lignans, a rapid procedure, based on 1H NMR spectroscopy, was developed for quantification of lignans in resins of C. atlantica.

  4. Quantitative determination of secondary metabolites in Cladina stellaris and other lichens by micellar electrokinetic chromatography.

    PubMed

    Falk, Adelia; Green, Thomas K; Barboza, Perry

    2008-02-22

    The quantitative determination of usnic acid (UA), perlatolic acid (PA), and atranorin (AT) in Alaska lichens by micellar electrokinetic chromatography (MEKC) is reported. The background electrolyte (BGE) included sodium docecyl sulfate (SDS), and beta-cyclodextrin (beta-CD) in a high-pH borate buffer. The presence of beta-CD in the buffer significantly decreases peak width, especially for UA, as it decreases migration time for both UA and PA. Linear calibration curves for UA, PA, and AT were established using an internal standard of benzoic acid (BA). Concentration limits of detection (cLODs) are 2.5, 2.2 and 2.0microg/mL (S/N 3) for UA, PA, and AT, respectively. Dry samples of lichen were extracted at room temperature with acetone for 24h in the presence of BA as internal standard. Recoveries of UA from spiked samples ranged from 92 to 98%. Amounts of UA and PA in the lichen samples ranged from 0.28 to 1.7% dry weight and 0.02 to 0.23%, respectively. PMID:18207149

  5. [Quantitative determination of trace heavy metals in polluted soil by laser induced breakdown spectroscopy].

    PubMed

    Wu, Wen-Tao; Ma, Xiao-Hong; Zhao, Hua-Feng; Zheng, Ze-Ke; Zhang, Min; Liag, Yan-Biao

    2011-02-01

    Based on laser induced breakdown spectroscopy data analysis, a quantitative determination method of elements was introduced. This method takes the advantage of Fourier analysis to study different part of the spectrum-white noise, thermal radiation and emission spectral lines. Through band-pass filter, emission spectral lines and most noise were separated, only a little white noise was missed. By calculated similarity between detected line and its corresponding unit characteristic line, convolution intensity was introduced to measure the detected line intensity. With Cu 324.75 nm as the analysis line and Ti 337.28 nm as the base line, the way of separating spectrum mentioned before was carried out to detect trace Cu pollution in soil. Results contained two bright spots. First, The correlation coefficient between Cu concentration and the spectral intensity was up to 0.997 9. Last but not the least, the detectability of Cu in soil was 44 mg x kg(-1), which is below the trace element thresholds for Class 2 soil used in the environmental quality standard for soil in China. At the same time, the relative error is always below 10%. Therefore the feasibility of this method for portable and instant detection of heavy metals in polluted soil was verified.

  6. Optimized external IR reflection spectroscopy for quantitative determination of borophosphosilicate glass parameters

    SciTech Connect

    Zhang, L.; Franke, J.E.; Niemczyk, T.M.; Haaland, D.M.

    1997-02-01

    Infrared (IR) external reflection spectroscopy has been optimized for the quantitative determination of composition and film thickness of borophosphosilicate glass (BPSG) deposited on silicon wafer substrates. The precision of the partial least-squares calibrations for boron and phosphorus contents and thin-film thickness were measured as the cross-validated standard error of prediction statistic. The results showed that BPSG IR reflection spectra collected over a wide range of incident IR radiation angles (15{degree}, 25{degree}, 45{degree}, and 60{degree}) can be used for the simultaneous quantification of these three BPSG parameters. When high angles of incidence were employed, the measurement was found to be more sensitive to small errors in the angle of incidence. The polarization state of the incident IR radiation did not noticeably affect the prediction of the three calibrated BPSG parameters. The results achieved in this study provide guidelines for at-line process monitoring and quality control of BPSG thin films used in the fabrication of microelectronic devices. {copyright} {ital 1997} {ital Society for Applied Spectroscopy}

  7. Fuzzy hidden Markov chains segmentation for volume determination and quantitation in PET

    NASA Astrophysics Data System (ADS)

    Hatt, M.; Lamare, F.; Boussion, N.; Turzo, A.; Collet, C.; Salzenstein, F.; Roux, C.; Jarritt, P.; Carson, K.; Cheze-LeRest, C.; Visvikis, D.

    2007-07-01

    Accurate volume of interest (VOI) estimation in PET is crucial in different oncology applications such as response to therapy evaluation and radiotherapy treatment planning. The objective of our study was to evaluate the performance of the proposed algorithm for automatic lesion volume delineation; namely the fuzzy hidden Markov chains (FHMC), with that of current state of the art in clinical practice threshold based techniques. As the classical hidden Markov chain (HMC) algorithm, FHMC takes into account noise, voxel intensity and spatial correlation, in order to classify a voxel as background or functional VOI. However the novelty of the fuzzy model consists of the inclusion of an estimation of imprecision, which should subsequently lead to a better modelling of the 'fuzzy' nature of the object of interest boundaries in emission tomography data. The performance of the algorithms has been assessed on both simulated and acquired datasets of the IEC phantom, covering a large range of spherical lesion sizes (from 10 to 37 mm), contrast ratios (4:1 and 8:1) and image noise levels. Both lesion activity recovery and VOI determination tasks were assessed in reconstructed images using two different voxel sizes (8 mm3 and 64 mm3). In order to account for both the functional volume location and its size, the concept of % classification errors was introduced in the evaluation of volume segmentation using the simulated datasets. Results reveal that FHMC performs substantially better than the threshold based methodology for functional volume determination or activity concentration recovery considering a contrast ratio of 4:1 and lesion sizes of <28 mm. Furthermore differences between classification and volume estimation errors evaluated were smaller for the segmented volumes provided by the FHMC algorithm. Finally, the performance of the automatic algorithms was less susceptible to image noise levels in comparison to the threshold based techniques. The analysis of both

  8. Quantitative determination of pairing interactions for high-temperature superconductivity in cuprates.

    PubMed

    Bok, Jin Mo; Bae, Jong Ju; Choi, Han-Yong; Varma, Chandra M; Zhang, Wentao; He, Junfeng; Zhang, Yuxiao; Yu, Li; Zhou, X J

    2016-03-01

    A profound problem in modern condensed matter physics is discovering and understanding the nature of fluctuations and their coupling to fermions in cuprates, which lead to high-temperature superconductivity and the invariably associated strange metal state. We report the quantitative determination of normal and pairing self-energies, made possible by laser-based angle-resolved photoemission measurements of unprecedented accuracy and stability. Through a precise inversion procedure, both the effective interactions in the attractive d-wave symmetry and the repulsive part in the full symmetry are determined. The latter is nearly angle-independent. Near T c, both interactions are nearly independent of frequency and have almost the same magnitude over the complete energy range of up to about 0.4 eV, except for a low-energy feature at around 50 meV that is present only in the repulsive part, which has less than 10% of the total spectral weight. Well below T c, they both change similarly, with superconductivity-induced features at low energies. Besides finding the pairing self-energy and the attractive interactions for the first time, these results expose the central paradox of the problem of high T c: how the same frequency-independent fluctuations can dominantly scatter at angles ±π/2 in the attractive channel to give d-wave pairing and lead to angle-independent repulsive scattering. The experimental results are compared with available theoretical calculations based on antiferromagnetic fluctuations, the Hubbard model, and quantum-critical fluctuations of the loop-current order.

  9. Quantitative determination of pairing interactions for high-temperature superconductivity in cuprates.

    PubMed

    Bok, Jin Mo; Bae, Jong Ju; Choi, Han-Yong; Varma, Chandra M; Zhang, Wentao; He, Junfeng; Zhang, Yuxiao; Yu, Li; Zhou, X J

    2016-03-01

    A profound problem in modern condensed matter physics is discovering and understanding the nature of fluctuations and their coupling to fermions in cuprates, which lead to high-temperature superconductivity and the invariably associated strange metal state. We report the quantitative determination of normal and pairing self-energies, made possible by laser-based angle-resolved photoemission measurements of unprecedented accuracy and stability. Through a precise inversion procedure, both the effective interactions in the attractive d-wave symmetry and the repulsive part in the full symmetry are determined. The latter is nearly angle-independent. Near T c, both interactions are nearly independent of frequency and have almost the same magnitude over the complete energy range of up to about 0.4 eV, except for a low-energy feature at around 50 meV that is present only in the repulsive part, which has less than 10% of the total spectral weight. Well below T c, they both change similarly, with superconductivity-induced features at low energies. Besides finding the pairing self-energy and the attractive interactions for the first time, these results expose the central paradox of the problem of high T c: how the same frequency-independent fluctuations can dominantly scatter at angles ±π/2 in the attractive channel to give d-wave pairing and lead to angle-independent repulsive scattering. The experimental results are compared with available theoretical calculations based on antiferromagnetic fluctuations, the Hubbard model, and quantum-critical fluctuations of the loop-current order. PMID:26973872

  10. Reflection thermal diffuse x-ray scattering for quantitative determination of phonon dispersion relations

    SciTech Connect

    Mei, A. B.; Hellman, O.; Schlepuetz, C. M.; Rockett, A.; Chiang, T. -C.; Hultman, L.; Petrov, I.; Greene, J. E.

    2015-11-03

    Synchrotron reflection x-ray thermal diffuse scattering (TDS) measurements, rather than previously reported transmission TDS, are carried out at room temperature and analyzed using a formalism based upon second-order interatomic force constants and long-range Coulomb interactions to obtain quantitative determinations of MgO phonon dispersion relations (h) over bar omega(j) (q), phonon densities of states g((h) over bar omega), and isochoric temperature-dependent vibrational heat capacities cv (T). We use MgO as a model system for investigating reflection TDS due to its harmonic behavior as well as its mechanical and dynamic stability. Resulting phonon dispersion relations and densities of states are found to be in good agreement with independent reports from inelastic neutron and x-ray scattering experiments. Temperature-dependent isochoric heat capacities cv (T), computed within the harmonic approximation from (h) over bar omega(j) (q) values, increase with temperature from 0.4 x 10-4 eV/atom K at 100 K to 1.4 x 10-4 eV/atom K at 200 K and 1.9 x 10-4 eV/atom K at 300 K, in excellent agreement with isobaric heat capacity values cp (T) between 4 and 300 K. We anticipate that the experimental approach developed here will be valuable for determining vibrational properties of heteroepitaxial thin films since the use of grazing-incidence (θ ≲ θc where θc is the density-dependent critical angle) allows selective tuning of x-ray penetration depths to ≲ 10 nm.

  11. Reflection thermal diffuse x-ray scattering for quantitative determination of phonon dispersion relations

    DOE PAGES

    Mei, A. B.; Hellman, O.; Schlepuetz, C. M.; Rockett, A.; Chiang, T. -C.; Hultman, L.; Petrov, I.; Greene, J. E.

    2015-11-03

    Synchrotron reflection x-ray thermal diffuse scattering (TDS) measurements, rather than previously reported transmission TDS, are carried out at room temperature and analyzed using a formalism based upon second-order interatomic force constants and long-range Coulomb interactions to obtain quantitative determinations of MgO phonon dispersion relations (h) over bar omega(j) (q), phonon densities of states g((h) over bar omega), and isochoric temperature-dependent vibrational heat capacities cv (T). We use MgO as a model system for investigating reflection TDS due to its harmonic behavior as well as its mechanical and dynamic stability. Resulting phonon dispersion relations and densities of states are found tomore » be in good agreement with independent reports from inelastic neutron and x-ray scattering experiments. Temperature-dependent isochoric heat capacities cv (T), computed within the harmonic approximation from (h) over bar omega(j) (q) values, increase with temperature from 0.4 x 10-4 eV/atom K at 100 K to 1.4 x 10-4 eV/atom K at 200 K and 1.9 x 10-4 eV/atom K at 300 K, in excellent agreement with isobaric heat capacity values cp (T) between 4 and 300 K. We anticipate that the experimental approach developed here will be valuable for determining vibrational properties of heteroepitaxial thin films since the use of grazing-incidence (θ ≲ θc where θc is the density-dependent critical angle) allows selective tuning of x-ray penetration depths to ≲ 10 nm.« less

  12. Imaging of Lamb Waves in Plates for Quantitative Determination of Anisotropy using Photorefractive Dynamic Holography

    SciTech Connect

    Telschow, Kenneth Louis; Deason, Vance Albert; Schley, Robert Scott; Watson, Scott Marshall

    1998-06-01

    Anisotropic properties of sheet materials can be determined by measuring the propagation of Lamb waves in different directions. Electromagnetic acoustic transduction and laser ultrasonic methods provide noncontacting approaches that are often desired for application to industrial and processing environments. This paper describes a laser imaging approach utilizing the adaptive property of photorefractive materials to produce a real-time measurement of the antisymmetric Lamb wave mode in all directions simultaneously. Continuous excitation is employed enabling the data to be recorded and displayed by a CCD camera. Analysis of the image produces a direct quantitative determination of the phase velocity in all directions showing plate anisotropy in the plane. Many optical techniques for measuring ultrasonic motion at surfaces have been developed for use in applications such as vibration measurement and laser ultrasonics. Most of these methods have similar sensitivities and are based on time domain processing using homodyne, Fabry-Perot [1], and, more recently, photorefractive interferometry [2]. Generally, the methods described above do not allow measurement at more than one surface point simultaneously, requiring multiple beam movements and scanning in order to produce images of surface ultrasonic motion over a large area. Electronic speckle interferometry, including shearography, does provide images directly of vibrations over large surface areas. This method has proven very durable in the field for large displacement amplitudes of several wavelengths. In addition, a sensitivity of ë/3000 has been demonstrated under laboratory conditions [3]. Full-field imaging of traveling ultrasonic waves using digital shearography has been recently reported with sensitivity in the nanometer range [4]. With this method, optical interference occurs at the photodetector

  13. Quantitative determination of pairing interactions for high-temperature superconductivity in cuprates

    PubMed Central

    Bok, Jin Mo; Bae, Jong Ju; Choi, Han-Yong; Varma, Chandra M.; Zhang, Wentao; He, Junfeng; Zhang, Yuxiao; Yu, Li; Zhou, X. J.

    2016-01-01

    A profound problem in modern condensed matter physics is discovering and understanding the nature of fluctuations and their coupling to fermions in cuprates, which lead to high-temperature superconductivity and the invariably associated strange metal state. We report the quantitative determination of normal and pairing self-energies, made possible by laser-based angle-resolved photoemission measurements of unprecedented accuracy and stability. Through a precise inversion procedure, both the effective interactions in the attractive d-wave symmetry and the repulsive part in the full symmetry are determined. The latter is nearly angle-independent. Near Tc, both interactions are nearly independent of frequency and have almost the same magnitude over the complete energy range of up to about 0.4 eV, except for a low-energy feature at around 50 meV that is present only in the repulsive part, which has less than 10% of the total spectral weight. Well below Tc, they both change similarly, with superconductivity-induced features at low energies. Besides finding the pairing self-energy and the attractive interactions for the first time, these results expose the central paradox of the problem of high Tc: how the same frequency-independent fluctuations can dominantly scatter at angles ±π/2 in the attractive channel to give d-wave pairing and lead to angle-independent repulsive scattering. The experimental results are compared with available theoretical calculations based on antiferromagnetic fluctuations, the Hubbard model, and quantum-critical fluctuations of the loop-current order. PMID:26973872

  14. Combining Quantitative Genetic Footprinting and Trait Enrichment Analysis to Identify Fitness Determinants of a Bacterial Pathogen

    PubMed Central

    Wiles, Travis J.; Norton, J. Paul; Russell, Colin W.; Dalley, Brian K.; Fischer, Kael F.; Mulvey, Matthew A.

    2013-01-01

    Strains of Extraintestinal Pathogenic Escherichia c oli (ExPEC) exhibit an array of virulence strategies and are a major cause of urinary tract infections, sepsis and meningitis. Efforts to understand ExPEC pathogenesis are challenged by the high degree of genetic and phenotypic variation that exists among isolates. Determining which virulence traits are widespread and which are strain-specific will greatly benefit the design of more effective therapies. Towards this goal, we utilized a quantitative genetic footprinting technique known as transposon insertion sequencing (Tn-seq) in conjunction with comparative pathogenomics to functionally dissect the genetic repertoire of a reference ExPEC isolate. Using Tn-seq and high-throughput zebrafish infection models, we tracked changes in the abundance of ExPEC variants within saturated transposon mutant libraries following selection within distinct host niches. Nine hundred and seventy bacterial genes (18% of the genome) were found to promote pathogen fitness in either a niche-dependent or independent manner. To identify genes with the highest therapeutic and diagnostic potential, a novel Trait Enrichment Analysis (TEA) algorithm was developed to ascertain the phylogenetic distribution of candidate genes. TEA revealed that a significant portion of the 970 genes identified by Tn-seq have homologues more often contained within the genomes of ExPEC and other known pathogens, which, as suggested by the first axiom of molecular Koch's postulates, is considered to be a key feature of true virulence determinants. Three of these Tn-seq-derived pathogen-associated genes—a transcriptional repressor, a putative metalloendopeptidase toxin and a hypothetical DNA binding protein—were deleted and shown to independently affect ExPEC fitness in zebrafish and mouse models of infection. Together, the approaches and observations reported herein provide a resource for future pathogenomics-based research and highlight the diversity of

  15. Thin-layer chromatography of aluminium: quantitative densitometric determination of Fe2+, Ni2+, Cu2+, and Si4+.

    PubMed

    Mohamed Najar, P A; Chouhan, R N; Jeurkar, J U; Dolas, S D; Ramana Rao, K V

    2007-01-01

    Thin-layer chromatography in combination with scanning densitometry is used as a tool for the quantitative determination of some impurity and additive elements in aluminium. Microgram levels of iron, silicon, copper, nickel, titanium, magnesium, manganese, and zinc present in a high concentration aluminium matrix is detected, and selective separations of some of these elements are achieved on silica gel H layers developed with a mobile phase containing aqueous sodium chloride solution. The quantitative determination of iron, silicon, nickel, and copper are obtained from the densitometric evaluation of chromatograms and are compared with the respective optical emission spectral analytical data. PMID:17555634

  16. Accurate determination of ultra-trace impurities, including europium, in ultra-pure barium carbonate materials through inductively coupled plasma-tandem mass spectrometry

    NASA Astrophysics Data System (ADS)

    Wu, Shuchao; Zeng, Xiangcheng; Dai, Xuefeng; Hu, Yongping; Li, Gang; Zheng, Cunjiang

    2016-09-01

    Impurities, especially ultra-trace europium (Eu), in ultra-pure barium carbonate materials were accurately determined through inductively coupled plasma-tandem mass spectrometry (ICP-MS/MS). Two reaction modes, namely, mass shift (with O2 as reaction gas) and on-mass modes(with NH3/He and He as reaction gases), were extensively investigated using Eu+ as target analyte. The use of Eu+ → EuO2+, instead of Eu+ → EuO+, as ion pairs in mass shift mode eliminated polyatomic interferences based on Ba matrix ions (135Ba16O2+ on 151Eu16O+ and 137Ba16O2+ on 153Eu16O+). This procedure exhibited enhanced sensitivity and selectivity. When the ICP-MS/MS was operated in NH3 on-mass mode, Eu+ can be determined in its original mass in interference-free conditions because NH3 did not react with Eu+ but with BaO+ to form a neutral product (BaO). The two reaction modes, especially NH3 on mass mode, were validated to be accurate because their resultant isotope ratios of 153Eu/151Eu matched well with that of the natural abundance ratio. The proposed ICP-MS/MS method is a sensitive technique with a limit of detection as low as 2.0 ng L- 1 for 153Eu+. Compared with conventional single-quadrupole (SQ) ICP-MS, both NH3 on-mass mode and O2 mass shift mode in ICP-MS/MS can be used to accurately determine Eu+ in ultra-pure BaCO3 materials. The detected concentration of Eu+ was 4.0 ng L- 1 to 15 ng L- 1, with spiked recoveries ranging from 100%-110%. ICP-MS/MS was also used to eliminate polyatomic interferences, particularly Ba-based interferences, prior to measurement of Gd and Sm. Impurities, including Na, Mg, Al, K, Mn, Fe, Cr, Sr, and Cs, in ultra-pure BaCO3 materials were also determined using ICP-MS/MS in conventional SQ mode.

  17. Quantitative mineralogy of the Yukon River system: Changes with reach and season, and determining sediment provenance

    USGS Publications Warehouse

    Eberl, D.D.

    2004-01-01

    mineral dissolution during transport between Eagle and Pilot Station, a distance of over 2000 km. We estimate that approximately 3 wt% of the quartz, 15 wt% of the feldspar (1 wt% of the alkali and 25 wt% of the plagioclase), and 26 wt% of the carbonates (31 wt% of the calcite and 15 wt% of the dolomite) carried by the river dissolve in this reach. The mineralogies of the suspended sediments change with the season. For example, during the summer of 2002 the quartz content varied by 20 wt%, with a minimum in mid-summer. The calcite content varied by a similar amount, and had a maximum corresponding to the quartz minimum. These modes are related to the relative amount of sediment flowing from the White River system, which is relatively poor in quartz, but rich in carbonate minerals. Suspended total clay minerals varied by as much as 25 wt%, with maxima in mid July, and suspended feldspar varied up to 10 wt%. Suspended sediment data from the summers of 2001 and 2003 support the 2002 trends. A calculation technique was developed to determine theproportion of various sediment sources in a mixed sediment by unmixing its quantitative mineralogy. Results from this method indicate that at least three sediment sources can be identified quantitatively with good accuracy. With this technique, sediment mineralogies can be used to calculate the relative flux of sediment from different tributaries, thereby identifying sediment provenance.

  18. Rapid and Accurate Determination of Lipopolysaccharide O-Antigen Types in Klebsiella pneumoniae with a Novel PCR-Based O-Genotyping Method

    PubMed Central

    Shih, Yun-Jui; Cheong, Cheng-Man; Yi, Wen-Ching

    2015-01-01

    Klebsiella pneumoniae, a Gram-negative bacillus that causes life-threatening infections in both hospitalized patients and ambulatory persons, can be classified into nine lipopolysaccharide (LPS) O-antigen serotypes. The O-antigen type has important clinical and epidemiological significance. However, K. pneumoniae O serotyping is cumbersome, and the reagents are not commercially available. To overcome the limitations of conventional serotyping methods, we aimed to create a rapid and accurate PCR method for K. pneumoniae O genotyping. We sequenced the genetic determinants of LPS O antigen from serotypes O1, O2a, O2ac, O3, O4, O5, O8, O9, and O12. We established a two-step genotyping scheme, based on the two genomic regions associated with O-antigen biosynthesis. The first set of PCR primers, which detects alleles at the wzm-wzt loci of the wb gene cluster, distinguishes between O1/O2, O3, O4, O5, O8, O9, and O12. The second set of PCR primers, which detects alleles at the wbbY region, further differentiates between O1, O2a, and O2ac. We verified the specificity of O genotyping against the O-serotype reference strains. We then tested the sensitivity and specificity of O genotyping in K. pneumoniae, using the 56 K-serotype reference strains with known O serotypes determined by an inhibition enzyme-linked immunosorbent assay (iELISA). There is a very good correlation between the O genotypes and classical O serotypes. Three discrepancies were observed and resolved by nucleotide sequencing—all in favor of O genotyping. The PCR-based O genotyping, which can be easily performed in clinical and research microbiology laboratories, is a rapid and accurate method for determining the LPS O-antigen types of K. pneumoniae isolates. PMID:26719438

  19. Application of double-spike isotope dilution for the accurate determination of Cr(III), Cr(VI) and total Cr in yeast.

    PubMed

    Yang, Lu; Ciceri, Elena; Mester, Zoltán; Sturgeon, Ralph E

    2006-11-01

    A method is presented for the simultaneous determination of Cr(III) and Cr(VI) in yeast using species-specific double-spike isotope dilution (SSDSID) with anion-exchange liquid chromatography (LC) separation and sector field inductively coupled plasma mass spectrometric (SF-ICP-MS) detection. Total Cr is quantitated using ID SF-ICP-MS. Samples were digested on a hot plate at 95+/-2 degrees C for 6 h in an alkaline solution of 0.5 M NaOH and 0.28 M Na2CO3 for the determination of Cr(III) and Cr(VI), whereas microwave-assisted decomposition with HNO3 and H2O2 was used for the determination of total Cr. Concentrations of 2,014+/-16, 1,952+/-103 and 76+/-48 mg kg-1 (one standard deviation, n=4, 3, 3), respectively were obtained for total Cr, Cr(III) and Cr(VI) in the yeast sample. Significant oxidation of Cr(III) to Cr(VI) (24.2+/-7.6% Cr(III) oxidized, n=3) and reduction of Cr(VI) to Cr(III) (37.6+/-6.5% Cr(VI) reduced, n=3) occurred during alkaline extraction and subsequent chromatographic separation at pH 7. Despite this significant bidirectional redox transformation, quantitative recoveries for both Cr(III) and Cr(VI) were achieved using the SSDSID method. In addition, mass balance between total Cr and the sum of Cr(III) and Cr(VI) concentrations was achieved. Method detection limits of 0.3, 2 and 30 mg kg-1 were obtained for total Cr, Cr(VI) and Cr(III), respectively, based on a 0.2-g sub-sample.

  20. Infrared Spectroscopy as a Versatile Analytical Tool for the Quantitative Determination of Antioxidants in Agricultural Products, Foods and Plants

    PubMed Central

    Cozzolino, Daniel

    2015-01-01

    Spectroscopic methods provide with very useful qualitative and quantitative information about the biochemistry and chemistry of antioxidants. Near infrared (NIR) and mid infrared (MIR) spectroscopy are considered as powerful, fast, accurate and non-destructive analytical tools that can be considered as a replacement of traditional chemical analysis. In recent years, several reports can be found in the literature demonstrating the usefulness of these methods in the analysis of antioxidants in different organic matrices. This article reviews recent applications of infrared (NIR and MIR) spectroscopy in the analysis of antioxidant compounds in a wide range of samples such as agricultural products, foods and plants. PMID:26783838

  1. A simple and inclusive method to determine the habit plane in transmission electron microscope based on accurate measurement of foil thickness

    SciTech Connect

    Qiu, Dong Zhang, Mingxing

    2014-08-15

    A simple and inclusive method is proposed for accurate determination of the habit plane between bicrystals in transmission electron microscope. Whilst this method can be regarded as a variant of surface trace analysis, the major innovation lies in the improved accuracy and efficiency of foil thickness measurement, which involves a simple tilt of the thin foil about a permanent tilting axis of the specimen holder, rather than cumbersome tilt about the surface trace of the habit plane. Experimental study has been done to validate this proposed method in determining the habit plane between lamellar α{sub 2} plates and γ matrix in a Ti–Al–Nb alloy. Both high accuracy (± 1°) and high precision (± 1°) have been achieved by using the new method. The source of the experimental errors as well as the applicability of this method is discussed. Some tips to minimise the experimental errors are also suggested. - Highlights: • An improved algorithm is formulated to measure the foil thickness. • Habit plane can be determined with a single tilt holder based on the new algorithm. • Better accuracy and precision within ± 1° are achievable using the proposed method. • The data for multi-facet determination can be collected simultaneously.

  2. Accurate approach for determining fresh-water carbonate (H2CO3(*)) alkalinity, using a single H3PO4 titration point.

    PubMed

    Birnhack, Liat; Sabach, Sara; Lahav, Ori

    2012-10-15

    A new, simple and accurate method is introduced for determining H(2)CO(3)(*) alkalinity in fresh waters dominated by the carbonate weak-acid system. The method relies on a single H(3)PO(4) dosage and two pH readings (acidic pH value target: pH~4.0). The computation algorithm is based on the concept that the overall alkalinity mass of a solution does not change upon the addition of a non-proton-accepting species. The accuracy of the new method was assessed batch-wise with both synthetic and actual tap waters and the results were compared to those obtained from two widely used alkalinity analysis methods (titration to pH~4.5 and the Gran titration method). The experimental results, which were deliberately obtained with simple laboratory equipment (glass buret, general-purpose pH electrode, magnetic stirrer) proved the method to be as accurate as the conventional methods at a wide range of alkalinity values (20-400 mg L(-1) as CaCO(3)). Analysis of the relative error attained in the proposed method as a function of the target (acidic) pH showed that at the range 4.0

  3. ANSI/ASHRAE/IES Standard 90.1-2010 Final Determination Quantitative Analysis

    SciTech Connect

    Halverson, Mark A.; Rosenberg, Michael I.; Liu, Bing

    2011-10-31

    The U.S. Department of Energy (DOE) conducted a final quantitative analysis to assess whether buildings constructed according to the requirements of the American National Standards Institute (ANSI)/American Society of Heating, Refrigerating and Air-Conditioning Engineers (ASHRAE)/Illuminating Engineering Society of North America (IESNA) Standard 90.1-2010 (ASHRAE Standard 90.1-2010, Standard 90.1-2010, or 2010 edition) would result in energy savings compared with buildings constructed to ANSI/ASHRAE/IESNA Standard 90.1-2007(ASHRAE Standard 90.1-2007, Standard 90.1-2007, or 2007 edition). The final analysis considered each of the 109 addenda to ASHRAE Standard 90.1-2007 that were included in ASHRAE Standard 90.1-2010. All 109 addenda processed by ASHRAE in the creation of Standard 90.1-2010 from Standard 90.1-2007 were reviewed by DOE, and their combined impact on a suite of 16 building prototype models in 15 ASHRAE climate zones was considered. Most addenda were deemed to have little quantifiable impact on building efficiency for the purpose of DOE's final determination. However, out of the 109 addenda, 34 were preliminarily determined to have a measureable and quantifiable impact. A suite of 240 computer energy simulations for building prototypes complying with ASHRAE 90.1-2007 was developed. These prototypes were then modified in accordance with these 34 addenda to create a second suite of corresponding building simulations reflecting the same buildings compliant with Standard 90.1-2010. The building simulations were conducted using the DOE EnergyPlus building simulation software. The resulting energy use from the complete suite of 480 simulation runs was then converted to energy use intensity (EUI, or energy use per unit floor area) metrics (Site EUI, Primary EUI, and energy cost intensity [ECI]) results for each simulation. For each edition of the standard, these EUIs were then aggregated to a national basis for each prototype using weighting factors based on

  4. [Hyperspectral technology combined with CARS algorithm to quantitatively determine the SSC in Korla fragrant pear].

    PubMed

    Zhan, Bai-Shao; Ni, Jun-Hui; Li, Jun

    2014-10-01

    Hyperspectral imaging has large data volume and high dimensionality, and original spectra data includes a lot of noises and severe scattering. And, quality of acquired hyperspectral data can be influenced by non-monochromatic light, external stray light and temperature, which resulted in having some non-linear relationship between the acquired hyperspectral data and the predicted quality index. Therefore, the present study proposed that competitive adaptive reweighted sampling (CARS) algorithm is used to select the key variables from visible and near infrared hyperspectral data. The performance of CARS was compared with full spectra, successive projections algorithm (SPA), Monte Carlo-uninformative variable elimination (MC-UVE), genetic algorithm (GA) and GA-SPA (genetic algorithm-successive projections algorithm). Two hundred Korla fragrant pears were used as research object. SPXY algorithm was used to divided sample set to correction set with 150 samples and prediction set with 50 samples, respectively. Based on variables selected by different methods, linear PLS and nonlinear LS-SVM models were developed, respectively, and the performance of models was assessed using parameters r2, RMSEP and RPD. A comprehensive comparison found that GA, GA-SPA and CARS can effectively select the variables with strong and useful information. These methods can be used for selection of Vis-NIR hyperspectral data variables, particularly for CARS. LS-SVM model can obtain the best results for SSC prediction of Korla fragrant pear based on variables obtained from CARS method. r2, RMSEP and RPD were 0.851 2, 0.291 3 and 2.592 4, respectively. The study showed that CARS is an effectively hyperspectral variable selection method, and nonlinear LS-SVM model is more suitable than linear PLS model for quantitatively determining the quality of fra- grant pear based on hyperspectral information. PMID:25739220

  5. Quantitative 3D Determination of Radiosensitization by Bismuth-Based Nanoparticles.

    PubMed

    Alqathami, Mamdooh; Blencowe, Anton; Geso, Moshi; Ibbott, Geoffrey

    2016-03-01

    The nanoparticle-induced dose enhancement effect has been shown to improve the therapeutic efficacy of ionizing radiation in external beam radiotherapy. Whereas previous studies have focused on gold nanoparticles (AuNPs), no quantitative studies have been conducted to investigate the potential superiority of other high atomic number (Z) nanomaterials such as bismuth-based nanoparticles. The aims of this study were to experimentally validate and quantify the dose enhancement properties of commercially available bismuth-based nanoparticles (bismuth oxide (Bi2O3-NPs) and bismuth sulfide (Bi2S3-NPs)), and investigate their potential superiority over AuNPs in terms of radiation dose enhancement. Phantom cuvettes doped with and without nanoparticles where employed for measuring radiation dose enhancement produced from the interaction of radiation with metal nanoparticles. Novel 3D phantoms were employed to investigate the 3D spatial distribution of ionising radiation dose deposition. The phantoms were irradiated with kilovoltage and megavoltage X-ray beams and optical absorption changes were measured using a spectrophotometer and optical CT scanner. The radiation dose enhancement factors (DEFs) obtained for 50 nm diameter Bi2O3-NPs and AuNPs were 1.90 and 1.77, respectively, for 100 kV energy and a nanoparticle concentration of 0.5 mM. In addition, the DEFs of 5 nm diameter Bi2S3-NPs and AuNPs were determined to be 1.38 and 1.51, respectively, for 150 kV energy and a nanoparticle concentration of 0.25 mM. The results demonstrate that both bismuth-based nanoparticles can enhance the effects of radiation. For 6 MV energy the DEFs for all the investigated nanoparticles were lower (< 15%) than with kilovoltage energy. PMID:27280244

  6. Comparative qualitative and quantitative determination of alkaloids in narcotic and condiment Papaver somniferum cultivars.

    PubMed

    Frick, Susanne; Kramell, Robert; Schmidt, Jürgen; Fist, Anthony J; Kutchan, Toni M

    2005-05-01

    In the present study morphinan, tetrahydrobenzylisoquinoline, benzo[c]phenanthridine, and phthalideisoquinoline alkaloids were determined qualitatively and quantitatively by HPLC and LC-MS analysis in tissues of the Tasmanian Papaver somniferum L. elite cultivar C048-6-14-64. The data were compared with the results from the low-morphine cultivar "Marianne". In the elite cultivar, 91.2% of the latex alkaloids consist of the three pharmaceutically most valuable alkaloids: morphine, codeine, and thebaine. In the root system, the major alkaloids are sanguinarine/10-hydroxysanguinarine and dihydrosanguinarine/10-hydroxydihydrosanguinarine. In the stems and leaves of C048-6-14-64, the same alkaloids were measured as in the latex. In the stems, a gradient in relative total alkaloid content from the top downward toward the roots was observed. The concentration of morphine was decreasing toward the roots, whereas an increasing gradient from the upper to the lower stem parts was detected for codeine. The relative total alkaloid concentration in leaves remained constant; no gradient was observed. The cultivar "Marianne" displayed a shifted pattern of alkaloid accumulation and reduced levels of total alkaloid. In the condiment cultivar, 80.5% of the alkaloids of the latex consisted of the two phthalideisoquinoline alkaloids narcotoline and noscapine. Only 18.8% of the relative total alkaloid content were morphinan alkaloids. In contrast to the narcotic cultivar, in which the benzo[c]phenanthridines in roots dominated over the morphinan and tetrahydrobenzylisoquinoline alkaloids, the concentration of benzo[c]phenanthridines in "Marianne" was similar to that of morphinan and tetrahydrobenzylisoquinoline alkaloids. These data suggest a differential alkaloid regulation in each cultivar of P. somniferum.

  7. Quantitative determination and classification of energy drinks using near-infrared spectroscopy.

    PubMed

    Rácz, Anita; Héberger, Károly; Fodor, Marietta

    2016-09-01

    Almost a hundred commercially available energy drink samples from Hungary, Slovakia, and Greece were collected for the quantitative determination of their caffeine and sugar content with FT-NIR spectroscopy and high-performance liquid chromatography (HPLC). Calibration models were built with partial least-squares regression (PLSR). An HPLC-UV method was used to measure the reference values for caffeine content, while sugar contents were measured with the Schoorl method. Both the nominal sugar content (as indicated on the cans) and the measured sugar concentration were used as references. Although the Schoorl method has larger error and bias, appropriate models could be developed using both references. The validation of the models was based on sevenfold cross-validation and external validation. FT-NIR analysis is a good candidate to replace the HPLC-UV method, because it is much cheaper than any chromatographic method, while it is also more time-efficient. The combination of FT-NIR with multidimensional chemometric techniques like PLSR can be a good option for the detection of low caffeine concentrations in energy drinks. Moreover, three types of energy drinks that contain (i) taurine, (ii) arginine, and (iii) none of these two components were classified correctly using principal component analysis and linear discriminant analysis. Such classifications are important for the detection of adulterated samples and for quality control, as well. In this case, more than a hundred samples were used for the evaluation. The classification was validated with cross-validation and several randomization tests (X-scrambling). Graphical Abstract The way of energy drinks from cans to appropriate chemometric models.

  8. Quantitative determination and classification of energy drinks using near-infrared spectroscopy.

    PubMed

    Rácz, Anita; Héberger, Károly; Fodor, Marietta

    2016-09-01

    Almost a hundred commercially available energy drink samples from Hungary, Slovakia, and Greece were collected for the quantitative determination of their caffeine and sugar content with FT-NIR spectroscopy and high-performance liquid chromatography (HPLC). Calibration models were built with partial least-squares regression (PLSR). An HPLC-UV method was used to measure the reference values for caffeine content, while sugar contents were measured with the Schoorl method. Both the nominal sugar content (as indicated on the cans) and the measured sugar concentration were used as references. Although the Schoorl method has larger error and bias, appropriate models could be developed using both references. The validation of the models was based on sevenfold cross-validation and external validation. FT-NIR analysis is a good candidate to replace the HPLC-UV method, because it is much cheaper than any chromatographic method, while it is also more time-efficient. The combination of FT-NIR with multidimensional chemometric techniques like PLSR can be a good option for the detection of low caffeine concentrations in energy drinks. Moreover, three types of energy drinks that contain (i) taurine, (ii) arginine, and (iii) none of these two components were classified correctly using principal component analysis and linear discriminant analysis. Such classifications are important for the detection of adulterated samples and for quality control, as well. In this case, more than a hundred samples were used for the evaluation. The classification was validated with cross-validation and several randomization tests (X-scrambling). Graphical Abstract The way of energy drinks from cans to appropriate chemometric models. PMID:27531031

  9. Quantitative Determination of Dielectric Thin-Film Properties Using Infrared Emission Spectroscopy

    SciTech Connect

    Franke, J.E.; Haaland, D.M.; Niemczyk, T.M.; Zhang, S.

    1998-10-14

    We have completed an experimental study to investigate the use of infrared emission spectroscopy (IRES) for the quantitative analysis of borophosphosilicate glass (BPSG) thin films on silicon monitor wafers. Experimental parameters investigated included temperatures within the range used in the microelectronics industry to produce these films; hence the potential for using the IRES technique for real-time monitoring of the film deposition process has been evaluated. The film properties that were investigated included boron content, phosphorus content, film thickness, and film temperature. The studies were conducted over two temperature ranges, 125 to 225 *C and 300 to 400 *C. The later temperature range includes realistic processing temperatures for the chemical vapor deposition (CVD) of the BPSG films. Partial least squares (PLS) multivariate calibration methods were applied to spectral and film property calibration data. The cross-validated standard errors of prediction (CVSEP) fi-om the PLS analysis of the IRES spectraof21 calibration samples each measured at 6 temperatures in the 300 to 400 "C range were found to be 0.09 wt. `?40 for B, 0.08 wt. `%0 for P, 3.6 ~m for film thickness, and 1.9 *C for temperature. By lowering the spectral resolution fi-om 4 to 32 cm-l and decreasing the number of spectral scans fi-om 128 to 1, we were able to determine that all the film properties could be measured in less than one second to the precision required for the manufacture and quality control of integrated circuits. Thus, real-time in-situ monitoring of BPSG thin films formed by CVD deposition on Si monitor wafers is possible with the methods reported here.

  10. Determination of burn patient outcome by large-scale quantitative discovery proteomics

    PubMed Central

    Finnerty, Celeste C.; Jeschke, Marc G.; Qian, Wei-Jun; Kaushal, Amit; Xiao, Wenzhong; Liu, Tao; Gritsenko, Marina A.; Moore, Ronald J.; Camp, David G.; Moldawer, Lyle L.; Elson, Constance; Schoenfeld, David; Gamelli, Richard; Gibran, Nicole; Klein, Matthew; Arnoldo, Brett; Remick, Daniel; Smith, Richard D.; Davis, Ronald; Tompkins, Ronald G.; Herndon, David N.

    2013-01-01

    Objective Emerging proteomics techniques can be used to establish proteomic outcome signatures and to identify candidate biomarkers for survival following traumatic injury. We applied high-resolution liquid chromatography-mass spectrometry (LC-MS) and multiplex cytokine analysis to profile the plasma proteome of survivors and non-survivors of massive burn injury to determine the proteomic survival signature following a major burn injury. Design Proteomic discovery study. Setting Five burn hospitals across the U.S. Patients Thirty-two burn patients (16 non-survivors and 16 survivors), 19–89 years of age, were admitted within 96 h of injury to the participating hospitals with burns covering >20% of the total body surface area and required at least one surgical intervention. Interventions None. Measurements and Main Results We found differences in circulating levels of 43 proteins involved in the acute phase response, hepatic signaling, the complement cascade, inflammation, and insulin resistance. Thirty-two of the proteins identified were not previously known to play a role in the response to burn. IL-4, IL-8, GM-CSF, MCP-1, and β2-microglobulin correlated well with survival and may serve as clinical biomarkers. Conclusions These results demonstrate the utility of these techniques for establishing proteomic survival signatures and for use as a discovery tool to identify candidate biomarkers for survival. This is the first clinical application of a high-throughput, large-scale LC-MS-based quantitative plasma proteomic approach for biomarker discovery for the prediction of patient outcome following burn, trauma or critical illness. PMID:23507713

  11. Quantitative Analysis by Isotopic Dilution Using Mass Spectroscopy: The Determination of Caffeine by GC-MS.

    ERIC Educational Resources Information Center

    Hill, Devon W.; And Others

    1988-01-01

    Describes a laboratory technique for quantitative analysis of caffeine by an isotopic dilution method for coupled gas chromatography-mass spectroscopy. Discusses caffeine analysis and experimental methodology. Lists sample caffeine concentrations found in common products. (MVL)

  12. Accurate determination of one-bond heteronuclear coupling constants with “pure shift” broadband proton-decoupled CLIP/CLAP-HSQC experiments

    NASA Astrophysics Data System (ADS)

    Timári, István; Kaltschnee, Lukas; Kolmer, Andreas; Adams, Ralph W.; Nilsson, Mathias; Thiele, Christina M.; Morris, Gareth A.; Kövér, Katalin E.

    2014-02-01

    We report broadband proton-decoupled CLIP/CLAP-HSQC experiments for the accurate determination of one-bond heteronuclear couplings and, by extension, for the reliable measurement of small residual dipolar coupling constants. The combination of an isotope-selective BIRD(d) filter module with a non-selective 1H inversion pulse is employed to refocus proton-proton coupling evolution prior to the acquisition of brief chunks of free induction decay that are subsequently assembled to reconstruct the fully-decoupled signal evolution. As a result, the cross-peaks obtained are split only by the heteronuclear one-bond coupling along the F2 dimension, allowing coupling constants to be extracted by measuring simple frequency differences between singlet maxima. The proton decoupling scheme presented has also been utilized in standard HSQC experiments, resulting in a fully-decoupled pure shift correlation map with significantly improved resolution.

  13. Hydrolysis Studies and Quantitative Determination of Aluminum Ions Using [superscript 27]Al NMR: An Undergraduate Analytical Chemistry Experiment

    ERIC Educational Resources Information Center

    Curtin, Maria A.; Ingalls, Laura R.; Campbell, Andrew; James-Pederson, Magdalena

    2008-01-01

    This article describes a novel experiment focused on metal ion hydrolysis and the equilibria related to metal ions in aqueous systems. Using [superscript 27]Al NMR, the students become familiar with NMR spectroscopy as a quantitative analytical tool for the determination of aluminum by preparing a standard calibration curve using standard aluminum…

  14. Determination of Calcium in Cereal with Flame Atomic Absorption Spectroscopy: An Experiment for a Quantitative Methods of Analysis Course

    ERIC Educational Resources Information Center

    Bazzi, Ali; Kreuz, Bette; Fischer, Jeffrey

    2004-01-01

    An experiment for determination of calcium in cereal using two-increment standard addition method in conjunction with flame atomic absorption spectroscopy (FAAS) is demonstrated. The experiment is intended to introduce students to the principles of atomic absorption spectroscopy giving them hands on experience using quantitative methods of…

  15. The Quantitative Determination of Food Dyes in Powdered Drink Mixes: A High School or General Science Experiment

    ERIC Educational Resources Information Center

    Sigmann, Samuella B.; Wheeler, Dale E.

    2004-01-01

    The development of a simple spectro photometric method to quantitatively determine the quantity of FD&C color additives present in powdered drink mixes, are focused by the investigations. Samples containing single dyes of binary mixtures of dyes can be analyzed using this method.

  16. QUANTITATIVE ENZYME-LINKED IMMUNOSORBENT ASSAY FOR DETERMINATION OF POLYCHLORINATED BIPHENYLS IN ENVIRONMENTAL SOIL AND SEDIMENT SAMPLES

    EPA Science Inventory

    An enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of Aroclors 1242, 1248, 1254, and 1260 in soil and sediments was developed and its performance compared with that of gas chromatography (GC). The detection limits for Aroclors 1242 and 1248 in soil ar...

  17. Fuzzy hidden Markov chains segmentation for volume determination and quantitation in PET

    PubMed Central

    Hatt, Mathieu; Lamare, Frédéric; Boussion, Nicolas; Roux, Christian; Turzo, Alexandre; Cheze-Lerest, Catherine; Jarritt, Peter; Carson, Kathryn; Salzenstein, Fabien; Collet, Christophe; Visvikis, Dimitris

    2007-01-01

    Accurate volume of interest (VOI) estimation in PET is crucial in different oncology applications such as response to therapy evaluation and radiotherapy treatment planning. The objective of our study was to evaluate the performance of the proposed algorithm for automatic lesion volume delineation; namely the Fuzzy Hidden Markov Chains (FHMC), with that of current state of the art in clinical practice threshold based techniques. As the classical Hidden Markov Chain (HMC) algorithm, FHMC takes into account noise, voxel’s intensity and spatial correlation, in order to classify a voxel as background or functional VOI. However the novelty of the fuzzy model consists of the inclusion of an estimation of imprecision, which should subsequently lead to a better modelling of the “fuzzy” nature of the object on interest boundaries in emission tomography data. The performance of the algorithms has been assessed on both simulated and acquired datasets of the IEC phantom, covering a large range of spherical lesion sizes (from 10 to 37mm), contrast ratios (4:1 and 8:1) and image noise levels. Both lesion activity recovery and VOI determination tasks were assessed in reconstructed images using two different voxel sizes (8mm3 and 64mm3). In order to account for both the functional volume location and its size, the concept of % classification errors was introduced in the evaluation of volume segmentation using the simulated datasets. Results reveal that FHMC performs substantially better than the threshold based methodology for functional volume determination or activity concentration recovery considering a contrast ratio of 4:1 and lesion sizes of <28mm. Furthermore differences between classification and volume estimation errors evaluated were smaller for the segmented volumes provided by the FHMC algorithm. Finally, the performance of the automatic algorithms was less susceptible to image noise levels in comparison to the threshold based techniques. The analysis of both

  18. A quantitative comparative analysis of Advancement via Independent Determination (AVID) in Texas middle schools

    NASA Astrophysics Data System (ADS)

    Reed, Krystal Astra

    The "Advancement via Individual Determination (AVID) program was designed to provide resources and strategies that enable underrepresented minority students to attend 4-year colleges" (AVID Center, 2013, p. 2). These students are characterized as the forgotten middle in that they have high test scores, average-to-low grades, minority or low socioeconomic status, and will be first-generation college students (AVID, 2011). Research indicates (Huerta, Watt, & Butcher, 2013) that strict adherence to 11 program components supports success of students enrolled in AVID, and AVID certification depends on districts following those components. Several studies (AVID Center, 2013) have investigated claims about the AVID program through qualitative analyses; however, very few have addressed this program quantitatively. This researcher sought to determine whether differences existed between student achievement and attendance rates between AVID and non-AVID middle schools. To achieve this goal, the researcher compared eighth-grade science and seventh- and eighth-grade mathematics scores from the 2007 to 2011 Texas Assessment of Knowledge and Skills (TAKS) and overall attendance rates in demographically equivalent AVID and non-AVID middle schools. Academic Excellence Indicator System (AEIS) reports from the Texas Education Agency (TEA) were used to obtain 2007 to 2011 TAKS results and attendance information for the selected schools. The results indicated a statistically significant difference between AVID demonstration students and non-AVID students in schools with similar CI. No statistically significant differences were found on any component of the TAKS for AVID economically disadvantaged students. The mean scores indicated an achievement gap between non-AVID and AVID demonstration middle schools. The findings from the other three research questions indicated no statistically significant differences between AVID and non-AVID student passing rates on the seventh- and eighth

  19. Accurate determination of the thickness or mass per unit area of thin foils and single-crystal wafers for x-ray attenuation measurements

    NASA Astrophysics Data System (ADS)

    Tran, C. Q.; Chantler, C. T.; Barnea, Z.; de Jonge, M. D.

    2004-09-01

    The determination of the local mass per unit area m/A=∫ρdt and the thickness of a specimen is an important aspect of its characterization and is often required for material quality control in fabrication. We discuss common methods which have been used to determine the local thickness of thin specimens. We then propose an x-ray technique which is capable of determining the local thickness and the x-ray absorption profile of a foil or wafer to high accuracy. This technique provides an accurate integration of the column density which is not affected by the presence of voids and internal defects in the material. The technique is best suited to specimens with thickness substantially greater than the dimensions of the surface and void structure. We also show that the attenuation of an x-ray beam by a nonuniform specimen is significantly different from that calculated by using a simple linear average of the mass per unit area and quantify this effect. For much thinner specimens or in the presence of a very structured surface profile we propose a complementary technique capable of attaining high accuracy by the use of a secondary standard. The technique is demonstrated by absolute measurements of the x-ray mass attenuation coefficient of copper and silver.

  20. Accurate determination of the thickness or mass per unit area of thin foils and single-crystal wafers for x-ray attenuation measurements

    SciTech Connect

    Tran, C.Q.; Chantler, C.T.; Barnea, Z.; Jonge, M.D. de

    2004-09-01

    The determination of the local mass per unit area m/A={integral}{rho}dt and the thickness of a specimen is an important aspect of its characterization and is often required for material quality control in fabrication. We discuss common methods which have been used to determine the local thickness of thin specimens. We then propose an x-ray technique which is capable of determining the local thickness and the x-ray absorption profile of a foil or wafer to high accuracy. This technique provides an accurate integration of the column density which is not affected by the presence of voids and internal defects in the material. The technique is best suited to specimens with thickness substantially greater than the dimensions of the surface and void structure. We also show that the attenuation of an x-ray beam by a nonuniform specimen is significantly different from that calculated by using a simple linear average of the mass per unit area and quantify this effect. For much thinner specimens or in the presence of a very structured surface profile we propose a complementary technique capable of attaining high accuracy by the use of a secondary standard. The technique is demonstrated by absolute measurements of the x-ray mass attenuation coefficient of copper and silver.

  1. Development of a coal/water-slurry-fueled diesel engine for industrial cogeneration: Task 6. 0 Determination of accurate heat release diagrams and mechanical efficiency

    SciTech Connect

    Nydick, S.E.

    1987-02-01

    Purpose of the program was to foster the long-range development of a coal/water slurry fuel-fired, slow-speed, two-stroke diesel engine for efficient and economical power generation in the 8 to 30 MW range for use in industrial cogeneration applications. This report contains the results of Task 6 of the program, a detailed analysis of heat release diagrams and determination of the mechanical efficiency for a single-cylinder, slow-speed diesel test engine when operated on coal/water slurry and diesel fuels. A digitized technique was utilized to determine the cylinder pressure history. The results of the program showed that the averages of previously reported thermal efficiency values for operation on coal/water slurry fuels were very accurate determinations. In addition, the slight differences in thermal efficiency between diesel and slurry operation are most likely related to changes in the mechanical efficiency resulting from degradation of the piston ring/cylinder liner interface rather than changes in combustion efficiency. Our previous prediction that, at 50% water content, the efficiency of a coal/water slurry engine is comparable to a diesel engine, has been confirmed experimentally and explained analytically.

  2. Quantitative determination of sulfonamide residues in foods of animal origin by high-performance liquid chromatography with fluorescence detection.

    PubMed

    Stoev, G; Michailova, A

    2000-02-25

    An HPLC method with fluorescence detection is proposed for the quantitative determination of residues of ten of the most used sulfonamides as their derivatives. Sulfonamides were isolated from meat, mix meat and kidney with ethyl acetate (first extraction) and acetone (second extraction) and further purified by partitioning three times with water-methylene chloride. The recovery for mix meat spiked with 1, 5 and 10 microg/kg of sulfonamides averaged 64%, 68% and 75%, respectively. Limits of quantitation were 1 microg/kg for sulfaquinoxaline and 0.5 microg/kg for the remaining sulfonamides. PMID:10735284

  3. Quantitative determination of nebivolol from human plasma using liquid chromatography-tandem mass spectrometry.

    PubMed

    Nandania, Jatin; Rajput, S J; Contractor, Pritesh; Vasava, Pragnesh; Solanki, Bhavik; Vohra, Mohsin

    2013-04-01

    In the present work, a rapid, sensitive, specific, precise and accurate liquid chromatography-tandem mass spectrometry method for determination of nebivolol in human plasma was developed and validated with a large calibration curve range (50-5000 pg/mL) which can be used for routine drug analysis and bioequivalence studies. Liquid-liquid extraction method was used to extract the analyte from the human plasma. The separation was achieved using Waters symmetry, C18, 4.6 × 150 mm, 5 μm column with formic acid in water, 0.01%, v/v: Acetonitrile (40:60) as a mobile phase. A flow rate of 0.8 mL/min, no splitting and run time 2.00 min was used for the chromatographic analysis of nebivolol. Sensitivity of this method was found to be 30 pg/mL. The analyte was analyzed by mass spectrometry in the multiple reaction monitoring mode. A Turbo-Ion spray source was interfaced between the HPLC and triple quadrupole mass spectrometer (MDS Sciex API 4000). The precursor-product ion m/z 406.00-151.00 for nebivolol and m/z 410.20-151.00 for nebivolol-D4 were used for quantification of an analyte and its IS. The method was validated in terms of accuracy, precision, selectivity, absolute recovery, freeze-thaw stability, bench-top stability, dry extract stability, short and long term stock solution stability, wet extract stability and re-injection reproducibility. The within- and between-batch accuracy was found to lie within the range of 87.00-100.40% and within- and between-batch precision was obtained within the range 0.33-8.67%. The mean recovery of all three concentration levels for drug was obtained 67.67% where as the mean recovery of IS was 68.74%. The %RSD value at higher concentration and lower concentration in all stability experiments was within 15%. This method is free from ion suppression, ion enhancement and any type of abnormal ionization.

  4. Formation resistivity measurements from within a cased well used to quantitatively determine the amount of oil and gas present

    DOEpatents

    Vail, III, William Banning

    2000-01-01

    Methods to quantitatively determine the separate amounts of oil and gas in a geological formation adjacent to a cased well using measurements of formation resistivity. The steps include obtaining resistivity measurements from within a cased well of a given formation, obtaining the porosity, obtaining the resistivity of formation water present, computing the combined amounts of oil and gas present using Archie's Equations, determining the relative amounts of oil and gas present from measurements within a cased well, and then quantitatively determining the separate amounts of oil and gas present in the formation. Resistivity measurements are obtained from within the cased well by conducting A.C. current from within the cased well to a remote electrode at a frequency that is within the frequency range of 0.1 Hz to 20 Hz.

  5. A Pure-Phase Homonuclear J-Modulated HMQC Experiment with Tilted Cross-Peak Patterns for an Accurate Determination of Homonuclear Coupling Constants

    NASA Astrophysics Data System (ADS)

    Koź Miń Ski, Wiktor

    1999-11-01

    A new HMQC-based experiment is presented which allows for an efficient determination of accurate homonuclear coupling-constant values. Pure absorption lineshapes with tilted cross-peak patterns are obtained by a combination of the active-coupling-pattern tilting (ACT) scheme with J-scaling. Characteristic features include separate heteronuclear echo and antiecho acquisition with a BIRDy pulse positioned before or after the t1 period, respectively, to refocus I-spin homonuclear coupling evolution. Additionally, due to the incorporation of J-scaling the relative spacing of the S-spin chemical-shift differences and I-spin homonuclear coupling splittings in the F1 domain is largely under experimental control. The most important advantage of the proposed method is that the I-spin homonuclear coupling evolution occurs simultaneously with the evolution of the heteronuclear zero and double-quantum coherences, which exhibit a slower transverse relaxation than I-spin single-quantum coherences. The effectiveness of the new sequence is demonstrated by a determination of the 3JHN,Hα couplings in a peptide sample. Additionally, the broadband property of the new sequence is verified with a sucrose sample.

  6. An optimized method for the accurate determination of patulin in apple products by isotope dilution-liquid chromatography/mass spectrometry.

    PubMed

    Seo, Miyeong; Kim, Byungjoo; Baek, Song-Yee

    2015-07-01

    Patulin, a mycotoxin produced by several molds in fruits, has been frequently detected in apple products. Therefore, regulatory bodies have established recommended maximum permitted patulin concentrations for each type of apple product. Although several analytical methods have been adopted to determine patulin in food, quality control of patulin analysis is not easy, as reliable certified reference materials (CRMs) are not available. In this study, as a part of a project for developing CRMs for patulin analysis, we developed isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS) as a higher-order reference method for the accurate value-assignment of CRMs. (13)C7-patulin was used as internal standard. Samples were extracted with ethyl acetate to improve recovery. For further sample cleanup with solid-phase extraction (SPE), the HLB SPE cartridge was chosen after comparing with several other types of SPE cartridges. High-performance liquid chromatography was performed on a multimode column for proper retention and separation of highly polar and water-soluble patulin from sample interferences. Sample extracts were analyzed by LC/MS/MS with electrospray ionization in negative ion mode with selected reaction monitoring of patulin and (13)C7-patulin at m/z 153→m/z 109 and m/z 160→m/z 115, respectively. The validity of the method was tested by measuring gravimetrically fortified samples of various apple products. In addition, the repeatability and the reproducibility of the method were tested to evaluate the performance of the method. The method was shown to provide accurate measurements in the 3-40 μg/kg range with a relative expanded uncertainty of around 1%.

  7. Accurate and self-consistent procedure for determining pH in seawater desalination brines and its manifestation in reverse osmosis modeling.

    PubMed

    Nir, Oded; Marvin, Esra; Lahav, Ori

    2014-11-01

    Measuring and modeling pH in concentrated aqueous solutions in an accurate and consistent manner is of paramount importance to many R&D and industrial applications, including RO desalination. Nevertheless, unified definitions and standard procedures have yet to be developed for solutions with ionic strength higher than ∼0.7 M, while implementation of conventional pH determination approaches may lead to significant errors. In this work a systematic yet simple methodology for measuring pH in concentrated solutions (dominated by Na(+)/Cl(-)) was developed and evaluated, with the aim of achieving consistency with the Pitzer ion-interaction approach. Results indicate that the addition of 0.75 M of NaCl to NIST buffers, followed by assigning a new standard pH (calculated based on the Pitzer approach), enabled reducing measured errors to below 0.03 pH units in seawater RO brines (ionic strength up to 2 M). To facilitate its use, the method was developed to be both conceptually and practically analogous to the conventional pH measurement procedure. The method was used to measure the pH of seawater RO retentates obtained at varying recovery ratios. The results matched better the pH values predicted by an accurate RO transport model. Calibrating the model by the measured pH values enabled better boron transport prediction. A Donnan-induced phenomenon, affecting pH in both retentate and permeate streams, was identified and quantified.

  8. Accurate determination of 2,4,6-trichloroanisole in wines at low parts per trillion by solid-phase microextraction followed by GC-ECD.

    PubMed

    Alzaga, Roberto; Ortiz, Laura; Sánchez-Baeza, Francisco; Marco, M-Pilar; Bayona, Josep Maria

    2003-06-01

    A headspace solid-phase microextraction (HS-SPME) procedure at 30 degrees C with a 100 microm PDMS fiber of a saturated NaCl solution stirred at 1100 rpm combined to GC-ECD for the 2,4,6-trichloroanisol (TCA) determination in wines has been developed. Due to the matrix complexity and ethanol absorption into the fiber, the internal standard selection was crucial to obtain unbiased results. Thus, matrix effects were observed when analyzing different types of Spanish wines (white, early, and vintage red wines) spiked with TCA at low concentration levels (i.e., <40 ng L(-)(1)). In contrast, the use of 2,4,6-tribromoanisole (TBA) as internal standard overcame these matrix effects, whereas the use of 2,4,6-trichlorophenyl ethyl ether led to inconsistent results. The developed HS-SPME-GC-ECD methodology reaches a limit of quantitation for TCA in wine within 2.9-18 ng L(-)(1), with a relative standard deviation of 2.5-13.4%, depending on the TCA concentration level and wine characteristics. This analytical method is comparable to the existing methodologies based on HS-SPME followed by GC-MS in terms of accuracy, precision, length of determination, and length of quantification; however, analysis cost is reduced.

  9. ANSI/ASHRAE/IES Standard 90.1-2013 Preliminary Determination: Quantitative Analysis

    SciTech Connect

    Halverson, Mark A.; Rosenberg, Michael I.; Wang, Weimin; Zhang, Jian; Mendon, Vrushali V.; Athalye, Rahul A.; Xie, YuLong; Hart, Reid; Goel, Supriya

    2014-03-01

    This report provides a preliminary quantitative analysis to assess whether buildings constructed according to the requirements of ANSI/ASHRAE/IES Standard 90.1-2013 would result in energy savings compared with buildings constructed to ANSI/ASHRAE/IES Standard 90.1-2010.

  10. A Colorimetric Analysis Experiment Not Requiring a Spectrophotometer: Quantitative Determination of Albumin in Powdered Egg White

    ERIC Educational Resources Information Center

    Charlton, Amanda K.; Sevcik, Richard S.; Tucker, Dorie A.; Schultz, Linda D.

    2007-01-01

    A general science experiment for high school chemistry students might serve as an excellent review of the concepts of solution preparation, solubility, pH, and qualitative and quantitative analysis of a common food product. The students could learn to use safe laboratory techniques, collect and analyze data using proper scientific methodology and…

  11. Validation of a Stability-Indicating Hydrophilic Interaction Liquid Chromatographic Method for the Quantitative Determination of Vitamin K3 (Menadione Sodium Bisulfite) in Injectable Solution Formulation

    PubMed Central

    Ghanem, Mashhour M.; Abu-Lafi, Saleh A.; Hallak, Hussein O.

    2013-01-01

    A simple, specific, accurate, and stability-indicating method was developed and validated for the quantitative determination of menadione sodium bisulfite in the injectable solution formulation. The method is based on zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) coupled with a photodiode array detector. The desired separation was achieved on the ZIC-HILIC column (250 mm × 4.6 mm, 5 μm) at 25°C temperature. The optimized mobile phase consisted of an isocratic solvent mixture of 200mM ammonium acetate (NH4AC) solution and acetonitrile (ACN) (20:80; v/v) pH-adjusted to 5.7 by glacial acetic acid. The mobile phase was fixed at 0.5 ml/min and the analytes were monitored at 261 nm using a photodiode array detector. The effects of the chromatographic conditions on the peak retention, peak USP tailing factor, and column efficiency were systematically optimized. Forced degradation experiments were carried out by exposing menadione sodium bisulfite standard and the injectable solution formulation to thermal, photolytic, oxidative, and acid-base hydrolytic stress conditions. The degradation products were well-resolved from the main peak and the excipients, thus proving that the method is a reliable, stability-indicating tool. The method was validated as per ICH and USP guidelines (USP34/NF29) and found to be adequate for the routine quantitative estimation of menadione sodium bisulfite in commercially available menadione sodium bisulfite injectable solution dosage forms. PMID:24106670

  12. A binary image reconstruction technique for accurate determination of the shape and location of metal objects in x-ray computed tomography.

    PubMed

    Wang, Jing; Xing, Lei

    2010-01-01

    The presence of metals in patients causes streaking artifacts in X-ray CT and has been recognized as a problem that limits various applications of CT imaging. Accurate localization of metals in CT images is a critical step for metal artifacts reduction in CT imaging and many practical applications of CT images. The purpose of this work is to develop a method of auto-determination of the shape and location of metallic object(s) in the image space. The proposed method is based on the fact that when a metal object is present in a patient, a CT image can be divided into two prominent components: high density metal and low density normal tissues. This prior knowledge is incorporated into an objective function as the regularization term whose role is to encourage the solution to take a form of two intensity levels. A computer simulation study and four experimental studies are performed to evaluate the proposed approach. Both simulation and experimental studies show that the presented algorithm works well even in the presence of complicated shaped metal objects. For a hexagonally shaped metal embedded in a water phantom, for example, it is found that the accuracy of metal reconstruction is within sub-millimeter.

  13. Selective and Accurate Determination Method of Propofol in Human Plasma by Mixed-Mode Cation Exchange Cartridge and GC-MS

    PubMed Central

    2016-01-01

    A gas chromatography-mass spectrometry (GC-MS) method for the determination of propofol in human plasma has been developed and validated. Propofol was extracted from human plasma by using mixed-mode cation exchange/reversed-phase (MCX) cartridges. As propofol easily volatilizes during concentration, 100% methanol was injected directly into GC-MS to elute propofol. Despite avoiding concentration process of the eluted solution, lower limit of quantization (LLOQ) of propofol was 25 ng/mL. The validated method exhibited good linearity (R2 = 0.9989) with accuracy and precision −5.8%~11.7% and 3.7%~11.6%, respectively. The other validation parameters, recovery and matrix effect, ranged from 96.6% to 99.4% and 95.3% to 101.4%, respectively. Propofol standard was quantified to evaluate possible loss due to the concentration processes, nitrogen gas and centrifugal vacuum. These two concentration processes resulted in notable decrease in the quantity of propofol, signifying avoiding any concentration processes during propofol quantification. Also, to confirm suitability of the developed method, authentic human plasma samples were analyzed. The selective assay method using MCX cartridge and GC-MS facilitated quantification of propofol in plasma sample accurately by preventing any losses due to the concentration processes.

  14. Selective and Accurate Determination Method of Propofol in Human Plasma by Mixed-Mode Cation Exchange Cartridge and GC-MS

    PubMed Central

    2016-01-01

    A gas chromatography-mass spectrometry (GC-MS) method for the determination of propofol in human plasma has been developed and validated. Propofol was extracted from human plasma by using mixed-mode cation exchange/reversed-phase (MCX) cartridges. As propofol easily volatilizes during concentration, 100% methanol was injected directly into GC-MS to elute propofol. Despite avoiding concentration process of the eluted solution, lower limit of quantization (LLOQ) of propofol was 25 ng/mL. The validated method exhibited good linearity (R2 = 0.9989) with accuracy and precision −5.8%~11.7% and 3.7%~11.6%, respectively. The other validation parameters, recovery and matrix effect, ranged from 96.6% to 99.4% and 95.3% to 101.4%, respectively. Propofol standard was quantified to evaluate possible loss due to the concentration processes, nitrogen gas and centrifugal vacuum. These two concentration processes resulted in notable decrease in the quantity of propofol, signifying avoiding any concentration processes during propofol quantification. Also, to confirm suitability of the developed method, authentic human plasma samples were analyzed. The selective assay method using MCX cartridge and GC-MS facilitated quantification of propofol in plasma sample accurately by preventing any losses due to the concentration processes. PMID:27597928

  15. Selective and Accurate Determination Method of Propofol in Human Plasma by Mixed-Mode Cation Exchange Cartridge and GC-MS.

    PubMed

    Pyo, Jae Sung

    2016-01-01

    A gas chromatography-mass spectrometry (GC-MS) method for the determination of propofol in human plasma has been developed and validated. Propofol was extracted from human plasma by using mixed-mode cation exchange/reversed-phase (MCX) cartridges. As propofol easily volatilizes during concentration, 100% methanol was injected directly into GC-MS to elute propofol. Despite avoiding concentration process of the eluted solution, lower limit of quantization (LLOQ) of propofol was 25 ng/mL. The validated method exhibited good linearity (R (2) = 0.9989) with accuracy and precision -5.8%~11.7% and 3.7%~11.6%, respectively. The other validation parameters, recovery and matrix effect, ranged from 96.6% to 99.4% and 95.3% to 101.4%, respectively. Propofol standard was quantified to evaluate possible loss due to the concentration processes, nitrogen gas and centrifugal vacuum. These two concentration processes resulted in notable decrease in the quantity of propofol, signifying avoiding any concentration processes during propofol quantification. Also, to confirm suitability of the developed method, authentic human plasma samples were analyzed. The selective assay method using MCX cartridge and GC-MS facilitated quantification of propofol in plasma sample accurately by preventing any losses due to the concentration processes. PMID:27597928

  16. A rapid and accurate method for determining protein content in dairy products based on asynchronous-injection alternating merging zone flow-injection spectrophotometry.

    PubMed

    Liang, Qin-Qin; Li, Yong-Sheng

    2013-12-01

    An accurate and rapid method and a system to determine protein content using asynchronous-injection alternating merging zone flow-injection spectrophotometry based on reaction between coomassie brilliant blue G250 (CBBG) and protein was established. Main merit of our approach is that it can avoid interferences of other nitric-compounds in samples, such as melamine and urea. Optimized conditions are as follows: Concentrations of CBBG, polyvinyl alcohol (PVA), NaCl and HCl are 150 mg/l, 30 mg/l, 0.1 mol/l and 1.0% (v/v), respectively; volumes of the sample and reagent are 150 μl and 30 μl, respectively; length of a reaction coil is 200 cm; total flow rate is 2.65 ml/min. The linear range of the method is 0.5-15 mg/l (BSA), its detection limit is 0.05 mg/l, relative standard deviation is less than 1.87% (n=11), and analytical speed is 60 samples per hour.

  17. Accurate determination of Curium and Californium isotopic ratios by inductively coupled plasma quadrupole mass spectrometry (ICP-QMS) in 248Cm samples for transmutation studies

    SciTech Connect

    Gourgiotis, A.; Isnard, H.; Aubert, M.; Dupont, E.; AlMahamid, I.; Cassette, P.; Panebianco, S.; Letourneau, A.; Chartier, F.; Tian, G.; Rao, L.; Lukens, W.

    2011-02-01

    The French Atomic Energy Commission has carried out several experiments including the mini-INCA (INcineration of Actinides) project for the study of minor-actinide transmutation processes in high intensity thermal neutron fluxes, in view of proposing solutions to reduce the radiotoxicity of long-lived nuclear wastes. In this context, a Cm sample enriched in {sup 248}Cm ({approx}97 %) was irradiated in thermal neutron flux at the High Flux Reactor (HFR) of the Laue-Langevin Institute (ILL). This work describes a quadrupole ICP-MS (ICP-QMS) analytical procedure for precise and accurate isotopic composition determination of Cm before sample irradiation and of Cm and Cf after sample irradiation. The factors that affect the accuracy and reproducibility of isotopic ratio measurements by ICP-QMS, such as peak centre correction, detector dead time, mass bias, abundance sensitivity and hydrides formation, instrumental background, and memory blank were carefully evaluated and corrected. Uncertainties of the isotopic ratios, taking into account internal precision of isotope ratio measurements, peak tailing, and hydrides formations ranged from 0.3% to 1.3%. This uncertainties range is quite acceptable for the nuclear data to be used in transmutation studies.

  18. In vivo skin analysis (INSA) for quantitative determination of lotion transfer to human skin.

    PubMed

    Ebrahimpour, Arman; Ullman, Alan H

    2009-01-01

    There is a need during the development of cosmetic and skin products for simple, quantitative, noninvasive measurements of product deposition onto skin. In this article we describe INSA (in vivo skin analysis) as such a method for measuring the amount of lotion transferred to the skin from tissue products. Using Fourier transform infrared spectroscopy with an attenuated total reflectance (ATR FT-IR) sampling accessory, we were able to quantify lotion levels on the arms of subjects in minutes.

  19. Quantitative performance of a quadrupole-orbitrap-MS in targeted LC-MS determinations of small molecules.

    PubMed

    Grund, Baptiste; Marvin, Laure; Rochat, Bertrand

    2016-05-30

    High-resolution mass spectrometry (HRMS) has been associated with qualitative and research analysis and QQQ-MS with quantitative and routine analysis. This view is now challenged and for this reason, we have evaluated the quantitative LC-MS performance of a new high-resolution mass spectrometer (HRMS), a Q-orbitrap-MS, and compared the results obtained with a recent triple-quadrupole MS (QQQ-MS). High-resolution full-scan (HR-FS) and MS/MS acquisitions have been tested with real plasma extracts or pure standards. Limits of detection, dynamic range, mass accuracy and false positive or false negative detections have been determined or investigated with protease inhibitors, tyrosine kinase inhibitors, steroids and metanephrines. Our quantitative results show that today's available HRMS are reliable and sensitive quantitative instruments and comparable to QQQ-MS quantitative performance. Taking into account their versatility, user-friendliness and robustness, we believe that HRMS should be seen more and more as key instruments in quantitative LC-MS analyses. In this scenario, most targeted LC-HRMS analyses should be performed by HR-FS recording virtually "all" ions. In addition to absolute quantifications, HR-FS will allow the relative quantifications of hundreds of metabolites in plasma revealing individual's metabolome and exposome. This phenotyping of known metabolites should promote HRMS in clinical environment. A few other LC-HRMS analyses should be performed in single-ion-monitoring or MS/MS mode when increased sensitivity and/or detection selectivity will be necessary.

  20. Accurate experimental determination of the isotope effects on the triple point temperature of water. I. Dependence on the 2H abundance

    NASA Astrophysics Data System (ADS)

    Faghihi, V.; Peruzzi, A.; Aerts-Bijma, A. T.; Jansen, H. G.; Spriensma, J. J.; van Geel, J.; Meijer, H. A. J.

    2015-12-01

    Variation in the isotopic composition of water is one of the major contributors to uncertainty in the realization of the triple point of water (TPW). Although the dependence of the TPW on the isotopic composition of the water has been known for years, there is still a lack of a detailed and accurate experimental determination of the values for the correction constants. This paper is the first of two articles (Part I and Part II) that address quantification of isotope abundance effects on the triple point temperature of water. In this paper, we describe our experimental assessment of the 2H isotope effect. We manufactured five triple point cells with prepared water mixtures with a range of 2H isotopic abundances encompassing widely the natural abundance range, while the 18O and 17O isotopic abundance were kept approximately constant and the 18O  -  17O ratio was close to the Meijer-Li relationship for natural waters. The selected range of 2H isotopic abundances led to cells that realised TPW temperatures between approximately  -140 μK to  +2500 μK with respect to the TPW temperature as realized by VSMOW (Vienna Standard Mean Ocean Water). Our experiment led to determination of the value for the δ2H correction parameter of A2H  =  673 μK / (‰ deviation of δ2H from VSMOW) with a combined uncertainty of 4 μK (k  =  1, or 1σ).

  1. Quantitative determination of the enantiomers of methadone in human plasma and saliva by chiral column chromatography coupled with mass spectrometric detection.

    PubMed

    George, Rani; Lobb, Michael; Haywood, Alison; Khan, Sohil; Hardy, Janet; Good, Phillip; Hennig, Stefanie; Norris, Ross

    2016-01-01

    Methadone is a potent lipophilic synthetic opioid that is effective in the treatment of cancer pain and perceived benefit in difficult pain control scenarios (especially in cases of neuropathic pain). The use of methadone in clinical practice is challenging however, due to the narrow therapeutic window and large inter- and intra-individual variability in therapeutic response. Quantitation of the enantiomers d- and l-methadone (d- and l-MTD) in plasma and saliva provides a basis for studying its pharmacokinetics in patients with cancer and for monitoring efficacy, toxicity and side-effects. This assay involves quantitation of the enantiomers of methadone using their respective deuterated internal standards, in plasma and saliva matrices with no impact of ion suppression in either matrix. The analytical recoveries of d- and l-MTD from the saliva collection devices (Salivette®) are optimised in this novel method with an accurate and simple extraction method employing dichloromethane. Optimal enantioselective separations were achieved using an α1-acid glycoprotein chiral stationary phase and triple quadrupole tandem mass spectrometer. Linearity was demonstrated over 0.05-1000µg/L for both enantiomers in plasma and in saliva with correlation coefficients greater than 0.998. The lower limit of quantitation (LLOQ) was determined to be 0.1µg/L in plasma and saliva for d- and l-MTD. Accuracy of the method ranges from 100% to 106% even at the LLOQ and total precision, expressed as the coefficient of variation, was between 0.2% and 4.4% for both analytes in both matrices. A simple one step extraction procedure resulted in recoveries greater than 95% for both analytes, at concentrations as low as 0.5µg/L, from the Salivette®. The validated method was applied successfully in 14 paired plasma and saliva samples obtained from adult patients with cancer pain receiving methadone.

  2. Quantitative High-Pressure Liquid Chromatography-Fluorescence Determination of Some Important Lower Fatty Acids in Lake Sediments

    PubMed Central

    Hordijk, Kees A.; Cappenberg, Thomas E.

    1983-01-01

    For the quantitative determination of traces of fatty acids in pore water, several gas and liquid chromatographic methods were tested and discussed. Direct determination by gas-liquid chromatography with the use of formic acid-saturated carrier gas was found to be the least laborious method, but it is only recommended for the determination of volatile acids such as acetate and higher homologs. For the determination of lactate and formate, a derivatization procedure is necessary. The determination of these acids as phenacyl or benzyl esters was complicated by contaminants in the reagents. For this reason, a high-pressure liquid chromatography procedure with 4-bromomethyl-7-methoxycoumarin as a fluorescent labeling reagent is preferred. With this method, lactic, acetic, and formic acids could be demonstrated simultaneously at the nanogram level in 5-ml samples. Profiles of these acids in the sediment of Lake Vechten were measured, and they showed correlations with sulfate-reducing and methanogenic bacterial activities. PMID:16346362

  3. Quantitative determination of amorphous nicardipine hydrochloride in long acting formula (NIC-LA) using light anhydrous silicic acid.

    PubMed

    Kohinata, Takeru; Fujii, Mitsuo; Nakamura, Souichiro; Hamada, Noritaka; Yonemochi, Etsuo; Terada, Katsuhide

    2004-12-01

    We investigated a method to quantitatively determine amorphous nicardipine hydrochloride (NIC) in the NIC-long acting formula (LA) model formulas prepared using NIC, light anhydrous silicic acid (LASA) and carboxymethylethylcellulose (CMEC). Consequently, since the quantity of total NIC in the formula can be determined by means of HPLC and crystal NIC can be determined by the differential scanning calorimetry (DSC) method because the heat of fusion (85.08 J/g) of NIC is constant and unaffected by excipients, we developed the HPLC-DSC method by which the quantity of amorphous NIC is calculated as the difference between the quantity of total NIC determined by HPLC and the quantity of crystal NIC determined by DSC. This practical HPLC-DSC method was confirmed to have good accuracy and reproducibility.

  4. Quantitative method for the determination of Iso-fludelone (KOS-1803) in human plasma by LC-MS/MS

    PubMed Central

    Christner, Susan M.; Parise, Robert A.; Levine, Erica D.; Rizvi, Naiyer A.; Gounder, Mrinal M.; Beumer, Jan H.

    2014-01-01

    Epothilones are relatively new tubulin-poison anticancer drugs. Iso-fludelone (KOS-1803) is a synthetic third generation epothilone drug discovered at Memorial Sloan Kettering Cancer Center, and currently in Phase I clinical trials. We report an LC-MS/MS assay for the sensitive, accurate and precise quantitation of Iso-fludelone in 0.2 mL of human plasma. Validation was performed according to FDA guidance. The assay comprised of KOS-1724 as the internal standard and an MTBE liquid-liquid extraction with a water wash step. Separation was achieved with an YMC-Pack ODS-AQ column and an isocratic mobile phase of 0.1% formic acid in acetonitrile and water (70:30, v/v) at 0.3 mL/min for 4 min. Chromatographic separation was followed by electrospray, positive-mode ionization tandem mass spectrometric detection in the multiple reaction monitoring (MRM) mode. The assay was linear from 0.1– 300 ng/mL and was accurate (−9.41–7.07%) and precise (1.03–13.7%) which fulfilled FDA criteria for validation. Recovery from plasma was 73.9–79.7% and ion suppression was negligible (−22.8 to −31.3%). Plasma freeze thaw stability (99.97–105.7%), stability for 11 months at −80 °C (94.93–107.9%), and stability for 6 h at room temperature (94.75–105.5%) were all acceptable. This assay is currently being applied to quantitate Iso-fludelone in clinical samples. PMID:25168219

  5. Critical considerations for the qualitative and quantitative determination of process-induced disorder in crystalline solids.

    PubMed

    Newman, Ann; Zografi, George

    2014-09-01

    Solid-state instabilities in crystalline solids arise during processing primarily because a certain level of structural disorder has been introduced into the crystal. Many physical instabilities appear to be associated with the recrystallization of molecules from these disordered regions, while chemical instabilities arise from sufficient molecular mobility to allow solid-state chemical reactivity. In this Commentary we discuss the various forms of structural disorder, processing which can produce disorder, the quantitative analysis of process-induced order, and strategies to limit disorder and its effects.

  6. Apparatus and method for quantitative determination of materials contained in fluids

    DOEpatents

    Radziemski, Leon J.; Cremers, David A.

    1985-01-01

    Apparatus and method for near real-time in-situ monitoring of particulates and vapors contained in fluids. Initial filtration of a known volume of the fluid sample is combined with laser-induced dielectric breakdown spectroscopy of the filter employed to obtain qualitative and quantitative information with high sensitivity. Application of the invention to monitoring of beryllium, beryllium oxide, or other beryllium-alloy dusts is demonstrated. Significant shortening of analysis time is achieved from those of the usual chemical techniques of analysis.

  7. Apparatus and method for quantitative determination of materials contained in fluids

    DOEpatents

    Radziemski, L.J.; Cremers, D.A.

    1982-09-07

    Apparatus and method for near real-time in-situ monitoring of particulates and vapors contained in fluids are described. Initial filtration of a known volume of the fluid sample is combined with laser-induced dielectric breakdown spectroscopy of the filter employed to obtain qualitative and quantitative information with high sensitivity. Application of the invention to monitoring of beryllium, beryllium oxide, or other beryllium-alloy dusts is shown. Significant shortening of analysis time is achieved from the usual chemical techniques of analysis.

  8. Simple methods for the qualitative identification and quantitative determination of macrolide antibiotics.

    PubMed

    Danielson, N D; Holeman, J A; Bristol, D C; Kirzner, D H

    1993-02-01

    Pyrolysis-gas chromatography is shown to be a rapid straightforward method for the qualitative differentiation of the macrolide antibiotics erythromycin, oleandomycin, troleandomycin, spiramycin and tylosin. Organic salts do not interfere and identification of erythromycin and troleandomycin in commercial products is viable. Spectrophotometric quantitation of these same five antibiotics after reaction with concentrated sulphuric acid is studied at about 470 nm. Reaction conditions such as acid concentration, time and temperature are provided. The sugar moieties of the antibiotics are proposed as the reactive sites. Detection limits are about 0.2-1.0 microg ml-1 [corrected] and analysis of pharmaceutical products should be possible.

  9. A quantitative method for determination of aflatoxin B in roasted corn.

    PubMed

    Shannon, G M; Shotwell, O L

    1975-07-01

    Roasting aflatoxin-contaminated corn will reduce toxin levels. A quantitative analysis for aflatoxin in roasted corn has been developed by modifying a cleanup technique for green coffee extracts approved as official first action by the AOAC. A chloroform extract is partially purified on a Florisil column, and thin layer chromatographic (TLC) plates are developed with methylene chloride-chloroform-isoamyl alcohol-formic acid (81+15+3+1). Recoveries average 101% and the sensitivity limit is 5 ppb aflatoxin B1. A 2-dimensional TLC procedure can also be used to separate the aflatoxins from background interferences. PMID:1150613

  10. Multiplex, quantitative, ligation-dependent probe amplification for determination of allergens in food.

    PubMed

    Mustorp, Stina L; Drømtorp, Signe M; Holck, Askild L

    2011-05-25

    Legislation requires labeling of foods containing allergenic ingredients. Here, we present a robust 10-plex quantitative and sensitive ligation-dependent probe amplification method, the allergen-multiplex ligation-dependent probe amplification (MLPA) method, for specific detection of eight allergens: sesame, soy, hazelnut, peanut, lupine, gluten, mustard, and celery. Ligated probes were amplified by polymerase chain reaction (PCR), and amplicons were detected using capillary electrophoresis. Quantitative results were obtained by comparing signals with an internal positive control. The limit of detection varied from approximately 5 to 400 gene copies, depending on the allergen. The method was tested using different foods spiked with mustard, celery, soy, or lupine flour in the 1-0.001% range. Depending on the allergen, sensitivities were similar or better than those obtained with qPCR. The allergen-MLPA method is modular and can be adapted by adding probe pairs for other allergens. The DNA-based allergen-MLPA method will constitute a complementary method to the traditional protein-based methods.

  11. Quantitative Determination of Fluorine Content in Blends of Polylactide (PLA)–Talc Using Near Infrared Spectroscopy

    PubMed Central

    Tamburini, Elena; Tagliati, Chiara; Bonato, Tiziano; Costa, Stefania; Scapoli, Chiara; Pedrini, Paola

    2016-01-01

    Near-infrared spectroscopy (NIRS) has been widely used for quantitative and/or qualitative determination of a wide range of matrices. The objective of this study was to develop a NIRS method for the quantitative determination of fluorine content in polylactide (PLA)-talc blends. A blending profile was obtained by mixing different amounts of PLA granules and talc powder. The calibration model was built correlating wet chemical data (alkali digestion method) and NIR spectra. Using FT (Fourier Transform)-NIR technique, a Partial Least Squares (PLS) regression model was set-up, in a concentration interval of 0 ppm of pure PLA to 800 ppm of pure talc. Fluorine content prediction (R2cal = 0.9498; standard error of calibration, SEC = 34.77; standard error of cross-validation, SECV = 46.94) was then externally validated by means of a further 15 independent samples (R2EX.V = 0.8955; root mean standard error of prediction, RMSEP = 61.08). A positive relationship between an inorganic component as fluorine and NIR signal has been evidenced, and used to obtain quantitative analytical information from the spectra. PMID:27490548

  12. Simultaneous quantitative determination of cinnamaldehyde and methyl eugenol from stem bark of Cinnamomum zeylanicum Blume using RP-HPLC.

    PubMed

    Gursale, Atish; Dighe, Vidya; Parekh, Guarang

    2010-01-01

    A simple, sensitive, and precise reversed-phase high performance liquid chromatographic (HPLC) method has been developed, validated and used for simultaneous quantitative determination of cinnamaldehyde and methyl eugenol from the methanolic extract of dried bark powder of Cinnamomum zeylanicum Blume (family Lauraceae). The ultrasonic extraction method was used for the extraction of these compounds. The reversed-phase HPLC analysis was carried out using a Intersil ODS-3V-C(18) (150 mm x 4.6 mm, 5 microm) column and a mobile phase comprising of methanol-acetonitrile-water in the volume ratio of 35:20:45, delivered at a flow rate of 1.0 cm(3)/min. The detection and quantitation of both the compounds was carried out at 221 nm.

  13. Quantitative determination of fatty acid chain composition in pork meat products by high resolution 1H NMR spectroscopy.

    PubMed

    Siciliano, Carlo; Belsito, Emilia; De Marco, Rosaria; Di Gioia, Maria Luisa; Leggio, Antonella; Liguori, Angelo

    2013-01-15

    High resolution (1)H NMR spectroscopy was proposed for the determination of the fatty acid chain profile of lipids in pork meat products during ripening. Two typical Mediterranean PDO salami produced in Calabria, a region in the Southern Italy, were chosen as a case of study. Quantitative NMR analysis provided the fatty acid chain profiles of total lipid extracts. The transesterification of total lipid extracts furnished FAME mixtures that enabled quantitation of fatty acid acyl chains in the acylglycerol and FFA portions. In all cases, oleyl chains were predominant, and high amounts of polyunsaturated fatty acid chains were observed. The proposed spectroscopic method allowed also the estimation of the most important nutritional parameters of dry fermented meat products.

  14. Quantitative determination of terbutaline and orciprenaline in human plasma by gas chromatography/negative ion chemical ionization/mass spectrometry.

    PubMed

    Leis, H J; Gleispach, H; Nitsche, V; Malle, E

    1990-06-01

    A method for the determination of unconjugated terbutaline and orciprenaline in human plasma is described. The assay is based on stable isotope dilution gas chromatography/negative ion chemical ionization/mass spectrometry. An inexpensive and rapid method for preparation of stable isotope labelled analogues as well as their use in quantitative gas chromatography/mass spectrometry is shown. A highly efficient sample work-up procedure with product recoveries of more than 95% is presented. The method developed permits quantitative measurement of terbutaline and orciprenaline in human plasma down to 100 pg ml-1, using 1 ml of sample. Plasma levels of terbutaline after oral administration of 5 mg of terbutaline sulphate were estimated. PMID:2357489

  15. Investigation of mass dependence effects for the accurate determination of molybdenum isotope amount ratios by MC-ICP-MS using synthetic isotope mixtures.

    PubMed

    Malinovsky, Dmitry; Dunn, Philip J H; Petrov, Panayot; Goenaga-Infante, Heidi

    2015-01-01

    Methodology for absolute Mo isotope amount ratio measurements by multicollector inductively coupled plasma-mass spectrometry (MC-ICP-MS) using calibration with synthetic isotope mixtures (SIMs) is presented. For the first time, synthetic isotope mixtures prepared from seven commercially available isotopically enriched molybdenum metal powders ((92)Mo, (94)Mo, (95)Mo, (96)Mo, (97)Mo, (98)Mo, and (100)Mo) are used to investigate whether instrumental mass discrimination of Mo isotopes in MC-ICP-MS is consistent with mass-dependent isotope distribution. The parent materials were dissolved and mixed as solutions to obtain mixtures with accurately known isotope amount ratios. The level of elemental impurities in the isotopically enriched molybdenum metal powders was quantified by ICP-MS by using both high-resolution and reaction cell instruments to completely resolve spectral interferences. The Mo isotope amount ratio values with expanded uncertainty (k = 2), determined by MC-ICP-MS for a high-purity Mo rod from Johnson Matthey, were as follows: (92)Mo/(95)Mo = 0.9235(9), (94)Mo/(95)Mo = 0.5785(8), (96)Mo/(95)Mo = 1.0503(9), (97)Mo/(95)Mo = 0.6033(6), (98)Mo/(95)Mo = 1.5291(20), and (100)Mo/(95)Mo = 0.6130(7). A full uncertainty budget for the measurements is presented which shows that the largest contribution to the uncertainty budget comes from correction for elemental impurities (∼51%), followed by the contribution from weighing operations (∼26 %). The atomic weight of molybdenum was calculated to be 95.947(2); the uncertainty in parentheses is expanded uncertainty with the coverage factor of 2. A particular advantage of the developed method is that calibration factors for all six Mo isotope amount ratios, involving the (95)Mo isotope, were experimentally determined. This allows avoiding any assumption on mass-dependent isotope fractions in MC-ICP-MS, inherent to the method of double spike previously used for Mo isotope amount ratio

  16. A method for volume determination of the orbit and its contents by high resolution axial tomography and quantitative digital image analysis.

    PubMed Central

    Cooper, W C

    1985-01-01

    The various congenital and acquired conditions which alter orbital volume are reviewed. Previous investigative work to determine orbital capacity is summarized. Since these studies were confined to postmortem evaluations, the need for a technique to measure orbital volume in the living state is presented. A method for volume determination of the orbit and its contents by high-resolution axial tomography and quantitative digital image analysis is reported. This procedure has proven to be accurate (the discrepancy between direct and computed measurements ranged from 0.2% to 4%) and reproducible (greater than 98%). The application of this method to representative clinical problems is presented and discussed. The establishment of a diagnostic system versatile enough to expand the usefulness of computerized axial tomography and polytomography should add a new dimension to ophthalmic investigation and treatment. Images FIGURE 8 FIGURE 9 FIGURE 10 A FIGURE 10 B FIGURE 11 A FIGURE 11 B FIGURE 12 FIGURE 13 FIGURE 14 FIGURE 15 FIGURE 16 FIGURE 17 FIGURE 18 FIGURE 19 FIGURE 20 FIGURE 21 FIGURE 22 FIGURE 23 FIGURE 24 FIGURE 25 FIGURE 26 A FIGURE 26 B FIGURE 27 FIGURE 28 FIGURE 29 FIGURE 30 FIGURE 31 FIGURE 32 PMID:3938582

  17. Development of a quantitative GC-FID method for the determination of stearoyl-lactylates (E481/482) in foods.

    PubMed

    Cucu, Tatiana; Shrestha, Kshitij; De Meulenaer, Bruno

    2014-01-01

    Sodium and calcium salts of stearoyl-lactylates (SLs) are food emulsifiers especially used in bread and bakery products to improve texture. They should be used at the lowest level at which the desired technological effect is achieved in a specific food category and at amounts not exceeding the maximums set by European Commission Regulation No. 1129/2011. In order to be able to evaluate whether these emulsifiers are used correctly but also to evaluate whether the commercial additive formulations comply with legislation, a quantitative GC-FID method was developed. An internal standard (nonadecanoyl-1-lactylate) was synthesized in-house and pure ester standards were isolated from commercial additive formulations. The method showed a limit of detection of 0.04 and a limit of quantification of 0.12 mg esters ml⁻¹. The commercial additive formulations analysed proved to be complex mixtures of free lactic and fatty acids together with only 50-60% esters. Besides SLs important amounts of palmitoyl-lactylates were present. Different food matrices (with low- and high-fat contents) were spiked with commercial SL formulations and recoveries ranged between 85% and 109%. Determination of SLs in commercial foods (such as bakery and bread) indicated that pre-treatment with amylase was essential to determine accurately the SL content due to the interaction of SL with the amylose. PMID:25268514

  18. Combination of differential growth at two different temperatures with a quantitative real-time polymerase chain reaction to determine temperature-sensitive phenotype of Mycoplasma synoviae.

    PubMed

    Shahid, Muhammad A; Ghorashi, Seyed A; Agnew-Crumpton, Rebecca; Markham, Philip F; Marenda, Marc S; Noormohammadi, Amir H

    2013-04-01

    Mycoplasma synoviae infections result in significant economic losses in the chicken and turkey industries. A commercially available live temperature-sensitive (ts (+)) vaccine strain MS-H has been found to be effective in controlling M. synoviae infections in commercial layer and broiler breeder farms in various countries, including Australia. Detection and differentiation of MS-H from field strains (ts (-)) and from ts (-) MS-H reisolates in vaccinated flocks is vital in routine flock status monitoring. At present microtitration is the only available technique to determine the ts phenotype of M. synoviae. This technique is time consuming and not amenable to automation. In the present study, a quantitative real-time polymerase chain reaction (Q-PCR) was combined with simultaneous culturing of M. synoviae at two different temperatures (33°C and 39.5°C) to determine the ts phenotype of 22 Australian M. synoviae strains/isolates. The M. synoviae type strain WVU-1853 was also included for comparison. A ratio of the copy numbers of the variable lipoprotein haemagglutinin (vlhA) gene at the two temperatures was calculated and a cut-off value was determined and used to delineate the ts phenotype. In all M. synoviae strains/isolates tested in this study, the ts phenotype determined using Q-PCR was in agreement with that determined using conventional microtitration. Combination of Q-PCR with differential growth at two different temperatures is a rapid, reliable and accurate technique that could be used as an effective tool in laboratories actively involved in ts phenotyping of M. synoviae strains/isolates.

  19. [Quantitative and qualitative determination of lactic acid in fecal filtrate in children].

    PubMed

    Filippskiĭ, G K; Tsaplin, Iu I

    1994-01-01

    The authors describe qualitative and quantitative methods for lactic acid assessment in children's feces. A total of 102 children with mainly lactose intolerance were examined. Benedict's test with feces filtrate, lactose tolerance test, and measurements of fecal carbohydrates were used in the diagnostic investigations. Mean lactic acid levels made up (mean +/- m) 33.3 +/- 3.6 mmol/l (6.3 +/- 1.0 in healthy infants on breast feeding and 6.0 +/- 1.3 mmol/l in those on mixed and formula feeding). Fecal carbohydrate levels were normal (below 5 g/l) in 12 infants, but lactic acid levels made up 67.8 +/- 6.1 mmol/l. Qualitative reaction to lactic acid in feces, starting from 44.4 mmol/l, was detected in 27% of infants. Differences in the values of healthy children and those with lactic carbohydrate intolerance were found significant (p < 0.001). PMID:7921814

  20. [Stich-plaque test--an economic method for quantitative determination of viruses].

    PubMed

    Waschke, K; Stäber, H

    1976-01-01

    An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID 50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity , accuracy, and reproductibility of this method are described and discussed.

  1. [Stick-plaque test--an economic method of quantitative determination of viruses].

    PubMed

    Waschke, K; Stäber, H

    1976-01-01

    An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity, accuracy, and reproductibility of this method are described and discussed.

  2. A method for quantitative determination of furanocoumarins in capsules and tablets of phytochemical preparations.

    PubMed

    Cardoso, Claudia Andréa Lima; Pires, Adriana Elias; Honda, Neli Kika

    2006-04-01

    A method for sample preparation and analysis by high-performance liquid chromatography with UV detection (HPLC-UV) was developed for analysis of psoralen, bergapten and 5-[3-(4,5-dihydro-5,5-dimethyl-4-oxo-2-furanyl)-butoxy]-7H-furo[3-2-g][1]benzopyran-7-one in capsules and tablets employed in Brazil for certain illnesses. The linearity, accuracy, the inter- and intra-day precision of the procedure were evaluated. Analytical curves for furanocoumarins were linear in the range of 1.0-50.0 microg/ml. The recoveries of the furanocoumarins in the products analyzed were 97.3-99.5%, and the percent coefficient of variation for the quantitative analysis of the furanocoumarins in the analyses was under 5%. For inter-equipment study gas chromatography (GC) was employed. PMID:16595942

  3. Quantitative determination of PAHs in biochar: a prerequisite to ensure its quality and safe application.

    PubMed

    Hilber, Isabel; Blum, Franziska; Leifeld, Jens; Schmidt, Hans-Peter; Bucheli, Thomas D

    2012-03-28

    Biochar is increasingly promoted as a beneficial soil conditioner. However, it may contain residues of polycyclic aromatic hydrocarbons (PAHs) as a result of its production by pyrolysis. To date, analytical methods to analyze PAHs in biochar quantitatively are hardly available. This study presents an optimized and validated procedure to quantify the 16 U.S. EPA PAHs in biochar. PAHs were best extracted with Soxhlet for 36 h using 100% toluene. Average absolute recoveries of isotope labeled internal standards used for each analyte from three different biochars ranged from 42% to 72%, and relative recoveries were between 71% and 105%. The limits of detection were biochar-dependent, but on average a factor of >50 lower than quantified PAH concentrations (9-355 mg kg(dry weight)(-1)). The established method prepares the ground for a harmonized protocol for PAH analysis of biochars, a necessity for biochar quality control, registration, and legislation.

  4. Quantitative Determination of Bandpasses for Producing Vegetation Indices from Recombined NEON Hyperspectral Imagery

    NASA Astrophysics Data System (ADS)

    Hulslander, D.

    2015-12-01

    Hyperspectral imaging systems can be used to produce spectral reflectance curves giving rich information about composition, relative abundances of materials, mixes and combinations. However, as each spectral return from these systems is a vector with several hundred elements, they can be very difficult to process and analyze, and problemeatic to compare within, across, and between datasets over time and space. Vegetation indices (e.g. NDVI, ARVI, EVI, et al) attempt to combine spectral features in to single-value scores. When derived from calibrated and atmospherically compensated reflectance data, these indices can be quantitatively compared. Historically, these indices have been calculated from multispectral sensor data. These sensors have a handful (4 to 16 or so) of bandbasses ranging from 20 nm to 200 nm FWHM covering specific spectral regions for a variety of reasons, including both intended applications and system limitations. Hyperspectral sensors, however, cover the spectrum with many, many narrow (5 to 10 nm) bandpasses. This allows for analyses using the full, detailed spectral curve, or combination of the bands in to regions by averaging or in to composites using transforms or other techniques. This raises the question of exactly which bands should be used and combined in what manner for ideally deriving well-known vegetation indices typically made from multispectral data. In this study we use derivatives and other curve and signal analysis techniques to analyze vegetation reflectance spectra to quantitatively define optimal bandpasses for several vegetation indices and combine the 5 nm hypserspectral bandpasses of the NEON Imaging Spectrometer to synthesize them.

  5. Quantitative determination of triterpenoid glycosides in Fatsia japonica Decne. & Planch. using high performance liquid chromatography.

    PubMed

    Ye, Xuewei; Yu, Siran; Lian, Xiao-Yuan; Zhang, Zhizhen

    2014-01-01

    Fatsia japonica Decne. & Planch. is a triterpenoid glycoside-rich herb with anti-inflammatory activity for the treatment of rheumatoid arthritis. A method for quantitative analysis of the complex triterpenoid glycosides in this medicinal plant has not been established so far. In this study, a high performance liquid chromatography (HPLC) method was developed for simultaneous qualification of 11 glycosides in F. japonica. The analysis was performed on an ODS-2 Hypersil column (250mm×4.6mm, 5μm) with a binary gradient mobile phase of water and acetonitrile. The established HPLC method was validated in terms of linearity, sensitivity, stability, precision, accuracy, and recovery. Results showed that this method had good linearity with R(2) at 0.99992-0.99999 in the test range of 0.04-9.00μg/μL. The limit of detection (LOD) and limit of quantification (LOQ) for the standard compounds were 0.013-0.020μg/μL and 0.040-0.060μg/μL. The relative standard deviations (RSDs%) of run variations were 0.83-1.40% for intra-day and 0.84-3.59% for inter-day. The analyzed compounds in the samples were stable for at least 36h, and the spike recoveries of the detected glycosides were 99.67-103.11%. The developed HPLC method was successfully applied for the measurements of the contents of 11 triterpenoid glycoside in different parts of F. japonica. Taken together, the HPLC method newly developed in this study could be used for qualitative and quantitative analysis of the bioactive triterpenoid glycosides in F. japonica and its products.

  6. The Quantitative Determination of Butylated Hydroxytoluene in Chewing Gum Using GC--MS

    ERIC Educational Resources Information Center

    Witter, A. E.

    2005-01-01

    The experiment to measure concentration of Photophysical Characterization(BHT) and determine percent recovery in chewing gum is described. The results demonstrated that over time, the concentration of BHT in the extract decreased owing to aerial oxidation.

  7. Liquid Chromatographic Determination of Nitroanilines: An Experiment for the Quantitative Analysis Laboratory.

    ERIC Educational Resources Information Center

    Cantwell, Frederick F.; Brown, David W.

    1981-01-01

    Describes a three-hour liquid chromatography experiment involving rapid separation of colored compounds in glass columns packed with a nonpolar absorbent. Includes apparatus design, sample preparation, experimental procedures, and advantages for this determination. (SK)

  8. Quantitative Chromatographic Determination of Dissolved Elemental Sulfur in the Non-aqueous Electrolyte for Lithium-Sulfur Batteries

    DOE PAGES

    Zheng, Dong; Yang, Xiao-Qing; Zhang, Xuran; Li, Chao; McKinnon, Meaghan E.; Sadok, Rachel G.; Qu, Deyu; Yu, Xiqian; Lee, Hung-Sui; Qu, Deyang

    2014-12-02

    A fast and reliable analytical method is reported for the quantitative determination of dissolved elemental sulfur in non-aqueous electrolytes for Li-S batteries. By using high performance liquid chromatography with a UV detector, the solubility of S in 12 different pure solvents and in 22 different electrolytes was determined. It was found that the solubility of elemental sulfur is dependent on the Lewis basicity, the polarity of solvents and the salt concentration in the electrolytes. In addition, the S content in the electrolyte recovered from a discharged Li-S battery was successfully determined by the proposed HPLC/UV method. Thus, the feasibility ofmore » the method to the online analysis for a Li-S battery is demonstrated. Interestingly, the S was found super-saturated in the electrolyte recovered from a discharged Li-S cell.« less

  9. Quantitative Chromatographic Determination of Dissolved Elemental Sulfur in the Non-aqueous Electrolyte for Lithium-Sulfur Batteries

    SciTech Connect

    Zheng, Dong; Yang, Xiao-Qing; Zhang, Xuran; Li, Chao; McKinnon, Meaghan E.; Sadok, Rachel G.; Qu, Deyu; Yu, Xiqian; Lee, Hung-Sui; Qu, Deyang

    2014-12-02

    A fast and reliable analytical method is reported for the quantitative determination of dissolved elemental sulfur in non-aqueous electrolytes for Li-S batteries. By using high performance liquid chromatography with a UV detector, the solubility of S in 12 different pure solvents and in 22 different electrolytes was determined. It was found that the solubility of elemental sulfur is dependent on the Lewis basicity, the polarity of solvents and the salt concentration in the electrolytes. In addition, the S content in the electrolyte recovered from a discharged Li-S battery was successfully determined by the proposed HPLC/UV method. Thus, the feasibility of the method to the online analysis for a Li-S battery is demonstrated. Interestingly, the S was found super-saturated in the electrolyte recovered from a discharged Li-S cell.

  10. A New Technique for Quantitative Determination of Dexamethasone in Pharmaceutical and Biological Samples Using Kinetic Spectrophotometric Method

    PubMed Central

    Akhoundi-Khalafi, Ali Mohammad; Shishehbore, Masoud Reza

    2015-01-01

    Dexamethasone is a type of steroidal medications that is prescribed in many cases. In this study, a new reaction system using kinetic spectrophotometric method for quantitative determination of dexamethasone is proposed. The method is based on the catalytic effect of dexamethasone on the oxidation of Orange G by bromate in acidic media. The change in absorbance as a criterion of the oxidation reaction progress was followed spectrophotometrically. To obtain the maximum sensitivity, the effective reaction variables were optimized. Under optimized experimental conditions, calibration graph was linear over the range 0.2–54.0 mg L−1. The calculated detection limit (3sb/m) was 0.14 mg L−1 for six replicate determinations of blank signal. The interfering effect of various species was also investigated. The present method was successfully applied for the determination of dexamethasone in pharmaceutical and biological samples satisfactorily. PMID:25737724

  11. Quantitative method of determining beryllium or a compound thereof in a sample

    DOEpatents

    McCleskey, T. Mark; Ehler, Deborah S.; John, Kevin D.; Burrell, Anthony K.; Collis, Gavin E.; Minogue, Edel M.; Warner, Benjamin P.

    2010-08-24

    A method of determining beryllium or a beryllium compound thereof in a sample, includes providing a sample suspected of comprising beryllium or a compound thereof, extracting beryllium or a compound thereof from the sample by dissolving in a solution, adding a fluorescent indicator to the solution to thereby bind any beryllium or a compound thereof to the fluorescent indicator, and determining the presence or amount of any beryllium or a compound thereof in the sample by measuring fluorescence.

  12. Quantitative method of determining beryllium or a compound thereof in a sample

    DOEpatents

    McCleskey, T. Mark; Ehler, Deborah S.; John, Kevin D.; Burrell, Anthony K.; Collis, Gavin E.; Minogue, Edel M.; Warner, Benjamin P.

    2006-10-31

    A method of determining beryllium or a beryllium compound thereof in a sample, includes providing a sample suspected of comprising beryllium or a compound thereof, extracting beryllium or a compound thereof from the sample by dissolving in a solution, adding a fluorescent indicator to the solution to thereby bind any beryllium or a compound thereof to the fluorescent indicator, and determining the presence or amount of any beryllium or a compound thereof in the sample by measuring fluorescence.

  13. A quantitative study for determination of sugar concentration using attenuated total reflectance terahertz (ATR-THz) spectroscopy

    NASA Astrophysics Data System (ADS)

    Suhandy, Diding; Suzuki, Tetsuhito; Ogawa, Yuichi; Kondo, Naoshi; Ishihara, Takeshi; Takemoto, Yuichiro

    2011-06-01

    The objective of our research was to use ATR-THz spectroscopy together with chemometric for quantitative study in food analysis. Glucose, fructose and sucrose are main component of sugar both in fresh and processed fruits. The use of spectroscopic-based method for sugar determination is well reported especially using visible, near infrared (NIR) and middle infrared (MIR) spectroscopy. However, the use of terahertz spectroscopy for sugar determination in fruits has not yet been reported. In this work, a quantitative study for sugars determination using attenuated total reflectance terahertz (ATR-THz) spectroscopy was conducted. Each samples of glucose, fructose and sucrose solution with different concentrations were prepared respectively and their absorbance spectra between wavenumber 20 and 450 cm-1 (between 0.6 THz and 13.5 THz) were acquired using a terahertz-based Fourier Transform spectrometer (FARIS-1S, JASCO Co., Japan). This spectrometer was equipped with a high pressure of mercury lamp as light source and a pyroelectric sensor made from deuterated L-alanine triglycine sulfate (DLTGS) as detector. Each spectrum was acquired using 16 cm-1 of resolution and 200 scans for averaging. The spectra of water and sugar solutions were compared and discussed. The results showed that increasing sugar concentration caused decreasing absorbance. The correlation between sugar concentration and its spectra was investigated using multivariate analysis. Calibration models for glucose, fructose and sucrose determination were developed using partial least squares (PLS) regression. The calibration model was evaluated using some parameters such as coefficient of determination (R2), standard error of calibration (SEC), standard error of prediction (SEP), bias between actual and predicted sugar concentration value and ratio prediction to deviation (RPD) parameter. The cross validation method was used to validate each calibration model. It is showed that the use of ATR

  14. A versatile phenomenological model for the S-shaped temperature dependence of photoluminescence energy for an accurate determination of the exciton localization energy in bulk and quantum well structures

    NASA Astrophysics Data System (ADS)

    Dixit, V. K.; Porwal, S.; Singh, S. D.; Sharma, T. K.; Ghosh, Sandip; Oak, S. M.

    2014-02-01

    Temperature dependence of the photoluminescence (PL) peak energy of bulk and quantum well (QW) structures is studied by using a new phenomenological model for including the effect of localized states. In general an anomalous S-shaped temperature dependence of the PL peak energy is observed for many materials which is usually associated with the localization of excitons in band-tail states that are formed due to potential fluctuations. Under such conditions, the conventional models of Varshni, Viña and Passler fail to replicate the S-shaped temperature dependence of the PL peak energy and provide inconsistent and unrealistic values of the fitting parameters. The proposed formalism persuasively reproduces the S-shaped temperature dependence of the PL peak energy and provides an accurate determination of the exciton localization energy in bulk and QW structures along with the appropriate values of material parameters. An example of a strained InAs0.38P0.62/InP QW is presented by performing detailed temperature and excitation intensity dependent PL measurements and subsequent in-depth analysis using the proposed model. Versatility of the new formalism is tested on a few other semiconductor materials, e.g. GaN, nanotextured GaN, AlGaN and InGaN, which are known to have a significant contribution from the localized states. A quantitative evaluation of the fractional contribution of the localized states is essential for understanding the temperature dependence of the PL peak energy of bulk and QW well structures having a large contribution of the band-tail states.

  15. HipMatch: an object-oriented cross-platform program for accurate determination of cup orientation using 2D-3D registration of single standard X-ray radiograph and a CT volume.

    PubMed

    Zheng, Guoyan; Zhang, Xuan; Steppacher, Simon D; Murphy, Stephen B; Siebenrock, Klaus A; Tannast, Moritz

    2009-09-01

    The widely used procedure of evaluation of cup orientation following total hip arthroplasty using single standard anteroposterior (AP) radiograph is known inaccurate, largely due to the wide variability in individual pelvic orientation relative to X-ray plate. 2D-3D image registration methods have been introduced for an accurate determination of the post-operative cup alignment with respect to an anatomical reference extracted from the CT data. Although encouraging results have been reported, their extensive usage in clinical routine is still limited. This may be explained by their requirement of a CAD model of the prosthesis, which is often difficult to be organized from the manufacturer due to the proprietary issue, and by their requirement of either multiple radiographs or a radiograph-specific calibration, both of which are not available for most retrospective studies. To address these issues, we developed and validated an object-oriented cross-platform program called "HipMatch" where a hybrid 2D-3D registration scheme combining an iterative landmark-to-ray registration with a 2D-3D intensity-based registration was implemented to estimate a rigid transformation between a pre-operative CT volume and the post-operative X-ray radiograph for a precise estimation of cup alignment. No CAD model of the prosthesis is required. Quantitative and qualitative results evaluated on cadaveric and clinical datasets are given, which indicate the robustness and the accuracy of the program. HipMatch is written in object-oriented programming language C++ using cross-platform software Qt (TrollTech, Oslo, Norway), VTK, and Coin3D and is transportable to any platform. PMID:19328585

  16. Qualitative and quantitative determination of ten iridoids and secoiridoids in Gentiana straminea Maxim. by LC-UV-ESI-MS.

    PubMed

    Wei, Shihu; Zhang, Peicheng; Feng, Xizhi; Kodama, Hiroyuki; Yu, Changyuan; Chen, Guang

    2012-01-01

    A simple and accurate HPLC-UV/MS method was developed for the simultaneous determination of ten iridoids and secoiridoids in the roots of Gentiana straminea Maxim. Separations were performed on a Kromasil-C18 column by gradient elution using methanol and water containing phosphoric acid. Analytes were identified by HPLC coupled with ESI-MS experiments. The chromatographic method was validated for selectivity, linearity, precision, limit of detection, limit of quantification, accuracy, and stability. The developed assay could be considered as a suitable quality control method for G. straminea and other "Qinjiao" herbs.

  17. Mesoscopic properties of interfacial ordering in amorphous germanium on Si(111) determined by quantitative digital image series matching.

    PubMed

    Thiel, K; Borgardt, N I; Plikat, B; Seibt, M

    2013-03-01

    For the quantitative characterization of atomic ordering in the transition region between crystalline and amorphous materials we have previously described a method based on averaging HREM images along the interface, simulation of averaged images with the use of the averaged projected potential approximation and determination of the atom arrangement by means of an iterative matching procedure for high-resolution focus series. In order to study mesoscopic properties of crystal induced ordering a fully quantitative procedure is developed in this work. For this purpose, the width of the averaging region is defined as a compromise providing necessary accuracy of calculations and desirable locality of characterization of the atom distribution. Fluctuations of the obtained atom distribution on the amorphous side of the interface are estimated by means a of special Monte-Carlo simulation technique. As a result, distribution functions obtained from different regions can be quantitatively compared and statistically significant differences can be identified and related to the atomic structure. The method is applied to investigate the near interfacial atom order at the interface between atomically flat crystalline Si(111) and amorphous Ge. It is shown that significant variations in the atomic density distribution occur on a 5-10nm scale for germanium atoms in the second and third atomic layer lying parallel to the interface.

  18. A new indicator and test paper for the semi-quantitative determination of carbaryl in traces in water

    SciTech Connect

    Rathore, H.S.; Saxena, S.K. )

    1990-01-01

    A chromatographic paper impregnated with sodium hydroxide (test paper), when spotted with a carbaryl solution and dipped in 2,6-dichloroquinone-4-chloroimine (Gibbs reagent) in benzene, turns blue with a violet tinge. The color formation is based upon the hydrolysis of carbaryl to give 1-naphthol which reacts with 2,6-dichloroquinone-4-chloroimine to produce indophenol that turns blue with a violet tinge in alkaline medium. The test paper can be successfully used for the detection and semi-quantitative determination of carbaryl at ppm levels in water.

  19. Quantitative determination of casein genetic variants in goat milk: Application in Girgentana dairy goat breed.

    PubMed

    Montalbano, Maria; Segreto, Roberta; Di Gerlando, Rosalia; Mastrangelo, Salvatore; Sardina, Maria Teresa

    2016-02-01

    The study was conducted to develop a high-performance liquid chromatographic (HPLC) method to quantify casein genetic variants (αs2-, β-, and κ-casein) in milk of homozygous individuals of Girgentana goat breed. For calibration experiments, pure genetic variants were extracted from individual milk samples of animals with known genotypes. The described HPLC approach was precise, accurate and highly suitable for quantification of goat casein genetic variants of homozygous individuals. The amount of each casein per allele was: αs2-casein A = 2.9 ± 0.8 g/L and F = 1.8 ± 0.4 g/L; β-casein C = 3.0 ± 0.8 g/L and C1 = 2.0 ± 0.7 g/L and κ-casein A = 1.6 ± 0.3 g/L and B = 1.1 ± 0.2 g/L. A good correlation was found between the quantities of αs2-casein genetic variants A and F, and β-casein C and C1 with other previously described method. The main important result was obtained for κ-casein because, till now, no data were available on quantification of single genetic variants for this protein. PMID:26304408

  20. Quantitative determination of casein genetic variants in goat milk: Application in Girgentana dairy goat breed.

    PubMed

    Montalbano, Maria; Segreto, Roberta; Di Gerlando, Rosalia; Mastrangelo, Salvatore; Sardina, Maria Teresa

    2016-02-01

    The study was conducted to develop a high-performance liquid chromatographic (HPLC) method to quantify casein genetic variants (αs2-, β-, and κ-casein) in milk of homozygous individuals of Girgentana goat breed. For calibration experiments, pure genetic variants were extracted from individual milk samples of animals with known genotypes. The described HPLC approach was precise, accurate and highly suitable for quantification of goat casein genetic variants of homozygous individuals. The amount of each casein per allele was: αs2-casein A = 2.9 ± 0.8 g/L and F = 1.8 ± 0.4 g/L; β-casein C = 3.0 ± 0.8 g/L and C1 = 2.0 ± 0.7 g/L and κ-casein A = 1.6 ± 0.3 g/L and B = 1.1 ± 0.2 g/L. A good correlation was found between the quantities of αs2-casein genetic variants A and F, and β-casein C and C1 with other previously described method. The main important result was obtained for κ-casein because, till now, no data were available on quantification of single genetic variants for this protein.

  1. Quantitative determination of total coliforms and Escherichia coli in marine waters with chromogenic and fluorogenic media.

    PubMed

    Geissler, K; Manafi, M; Amorós, I; Alonso, J L

    2000-02-01

    This study compared the performance of LMX(R) broth (LMX), Chromocult Coliform(R) agar (CC) and Chromocult Coliform agar plus cefsulodin (10 microg ml-1) (CC-CFS), with standard methods multiple tube fermentation (MTF), for the enumeration of total coliforms and Escherichia coli from marine recreational waters. LMX and CC are two media designed to concurrently detect total coliform (TC) bacteria and E. coli by the specific action of beta-galactosidase (total coliforms) and beta-glucuronidase (E. coli). Overall results for the TC test showed that LMX, CC and MTF recovered 2.63, 1.95 and 1.90 times as many TCs as CC-CFS, respectively. Data from the multiple range test showed significant differences (P < 0.05) between TC counts on CC-CFS and LMX. The traditional MTF was less sensitive for E. coli enumeration. However, there was no statistically significant differences between LMX, CC, CC-CFS and the MTF method for E. coli enumeration. Background interference was reduced on CC-CFS and the counts obtained reflected more accurately the number of TCs. Therefore, the contribution of beta-galactosidase positive, non coliform bacteria (Aeromonas spp. and Vibrio spp.) to TC counts should not be neglected.

  2. Quantitative and combinatory determination of in situ phosphorylation of tau and its FTDP-17 mutants.

    PubMed

    Kimura, Taeko; Hosokawa, Tomohisa; Taoka, Masato; Tsutsumi, Koji; Ando, Kanae; Ishiguro, Koichi; Hosokawa, Masato; Hasegawa, Masato; Hisanaga, Shin-Ichi

    2016-01-01

    Tau is hyperphosphorylated in the brains of patients with tauopathies, such as Alzheimer's disease and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). However, neither the mechanism of hyperphosphorylation nor its contribution to pathogenesis is known. We applied Phos-tag SDS-PAGE, a phosphoaffinity electrophoresis, to the analysis of tau phosphorylation in vitro by Cdk5, in cultured cells and in mouse brain. Here, we found that Cdk5-p25 phosphorylated tau in vitro at Ser404, Ser235, Thr205 and Ser202 in this order. In contrast in cultured cells, Ser404 was preferentially phosphorylated by Cdk5-p35, whereas Thr205 was not phosphorylated. Ser202 and Ser235 were phosphorylated by endogenous kinases. Tau exhibited ~12 phosphorylation isotypes in COS-7 cells with different combinations of phosphorylation at Thr181, Ser202, Thr231, Ser235 and Ser404. These phosphorylation sites were similar to tau phosphorylated in mouse brains. FTDP-17 tau with a mutation in the C-terminal region had different banding patterns, indicating a different phosphorylation pattern. In particular, it was clear that the R406W mutation causes loss of Ser404 phosphorylation. These results demonstrate the usefulness of the Phos-tag technique in the quantitative analysis of site-specific in vivo phosphorylation of tau and provide detailed information on in situ combinatory phosphorylation of tau. PMID:27641626

  3. Variables selection for quantitative determination of cotton content in textile blends by near infrared spectroscopy

    NASA Astrophysics Data System (ADS)

    Sun, Xu-dong; Zhou, Ming-xing; Sun, Yi-ze

    2016-07-01

    Investigations were initiated to develop near infrared (NIR) techniques coupled with variables selection method to rapidly measure cotton content in blend fabrics of cotton and polyester. Multiplicative scatter correction (MSC), smooth, first derivative (1Der), second derivative (2Der) and their combination were employed to preprocess the spectra. Monte Carlo uninformative variables elimination (MCUVE), successive projections algorithm (SPA), and genetic algorithm (GA) were performed comparatively to choose characteristic variables associated with cotton content distributions. One hundred and thirty-five and fifty-nine samples were used to calibrate models and assess the performance of the models, respectively. Through comparing the performance of partial least squares (PLS) regression models with new samples, the optimal model of cotton content was obtained with spectral pretreatment method of 2 Der-Smooth-MSC and variables selection method of MCUVE-SPA-PLS. The correlation coefficient of prediction (rp) and root mean square errors of prediction (RMSEP) were 0.988% and 2.100%, respectively. The results suggest that NIR technique combining with variables selection method of MCUVE-SPA has significant potential to quantitatively analyze cotton content in blend fabrics of cotton and polyester; moreover, it could indicate the related spectral contributions.

  4. Quantitative and combinatory determination of in situ phosphorylation of tau and its FTDP-17 mutants

    PubMed Central

    Kimura, Taeko; Hosokawa, Tomohisa; Taoka, Masato; Tsutsumi, Koji; Ando, Kanae; Ishiguro, Koichi; Hosokawa, Masato; Hasegawa, Masato; Hisanaga, Shin-ichi

    2016-01-01

    Tau is hyperphosphorylated in the brains of patients with tauopathies, such as Alzheimer’s disease and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). However, neither the mechanism of hyperphosphorylation nor its contribution to pathogenesis is known. We applied Phos-tag SDS-PAGE, a phosphoaffinity electrophoresis, to the analysis of tau phosphorylation in vitro by Cdk5, in cultured cells and in mouse brain. Here, we found that Cdk5-p25 phosphorylated tau in vitro at Ser404, Ser235, Thr205 and Ser202 in this order. In contrast in cultured cells, Ser404 was preferentially phosphorylated by Cdk5-p35, whereas Thr205 was not phosphorylated. Ser202 and Ser235 were phosphorylated by endogenous kinases. Tau exhibited ~12 phosphorylation isotypes in COS-7 cells with different combinations of phosphorylation at Thr181, Ser202, Thr231, Ser235 and Ser404. These phosphorylation sites were similar to tau phosphorylated in mouse brains. FTDP-17 tau with a mutation in the C-terminal region had different banding patterns, indicating a different phosphorylation pattern. In particular, it was clear that the R406W mutation causes loss of Ser404 phosphorylation. These results demonstrate the usefulness of the Phos-tag technique in the quantitative analysis of site-specific in vivo phosphorylation of tau and provide detailed information on in situ combinatory phosphorylation of tau. PMID:27641626

  5. Fast-Neutron Hodoscope at TREAT: Methods for Quantitative Determination of Fuel Dispersal

    SciTech Connect

    De Volci, A.; Fink, C. L.; Marsh, G. E.; Rhodes, E. A.; Stanford, G. S.

    1980-01-01

    Fuel-motion surveillance using the fast-neutron hodoscope in TREAT experiments has advanced from an initial role of providing time/location/velocity data to that of offering quantitative mass results. The material and radiation surroundings of tha test section contribute to intrinsic and instrumental effects upon hodoscope detectors that require detailed corrections. Depending upon the experiment, count rate compensation is usually required for deadtime, power level, nonlinear response, efficiency, background, and detector calibration. Depending on their magnitude and amenability to analytical and empirical treatment, systematic corrections may be needed for self-shielding, self-multiplication, self-attenuation, flux depression, and other effects. Current verified hodoscope response (for 1- to 7-pin fuel bundles) may be paramatrically characterized under optimum conditions by 1-ms time resolution; 0.25-mm lateral and 5-mm axial-motion displacement resolution; and 50-mg single-pin mass resolution. The experimental and theoretical foundation for this performance is given, with particular emphasis on the geometrical response function and the statistical limits of fuel-motion resolution. Comparisons are made with alternative diagnostic systems.

  6. Evaluation of Real-Time Quantitative Polymerase Chain Reaction (qPCR) to Determine Escherichia coli Concentrations at Two Lake Erie Beaches

    USGS Publications Warehouse

    Kephart, Christopher M.; Bushon, Rebecca N.

    2009-01-01

    During the recreational seasons of 2006 and 2007, the quantitative polymerase chain reaction (qPCR) method was used to determine Escherichia coli (E. coli) concentrations in samples from two Lake Erie beaches. Results from the qPCR method were compared to those obtained by traditional culturing on modified mTEC agar. Regression analysis showed strong, statistically significant correlations between results from the two methods for both years. Correlation coefficients at Edgewater and Villa Angela Beaches were 0.626 and 0.789 for 2006 and 0.667 and 0.829 for 2007, respectively. Linear regression analyses were done to determine how well E. coli concentrations could have been predicted from qPCR results. These hypothetical predictions were compared to the current practice of determining recreational water quality from E. coli concentrations determined for samples collected on the previous day. The qPCR method resulted in a greater percentage of correct predictions of water-quality exceedances than the current method for both beaches and both years. However, because regression equations differed somewhat between both sites and both years, the study did not result in any single relation reliable enough to use for actual real-time prediction of water-quality exceedances for either beach; therefore, a posterior analysis of data was done. Additional years of data may be needed to develop such a relation. Results from this study support the continued development and testing of a qPCR method for providing rapid and accurate estimates of E. coli concentrations for monitoring recreational water quality.

  7. Quantitative radiochemical methods for determination of the sources of natural radioactivity

    USGS Publications Warehouse

    Rosholt, J.N.

    1957-01-01

    Study of the state of equilibrium of any natural radioactive source requires determination of several key nuclides or groups of nuclides to find their contribution to the total amount of radioactivity. Alpha activity measured by scintillation counting is used for determination of protactinium-231, thorium-232, thorium-230, and radium-226. The chemical procedures for the separations of the specific elements are described, as well as the measurement techniques used to determine the abundances of the individual isotopes. To correct for deviations in the ore standards, an independent means of evaluating the efficiencies of the individual separations and measurements is used. The development of these methods of radiochemical analysis facilitates detailed investigation of the major sources of natural radioactivity.

  8. I. QUANTITATIVE DETERMINATION OF THE AMOUNT OF DNA PER NUCLEUS BY INTERFERENCE MICROSCOPY

    PubMed Central

    Bibbiani, Cecilia; Tongiani, Roberto; Viola-Magni, Maria Pia

    1969-01-01

    A method for the determination of the DNA content of isolated nuclei of different ploidy has been developed. It is based on measurement of the nuclear dry mass, with an integrating microinterferometer, before and after DNase treatment. The values found are slightly low, because, as indicated by biochemical determinations, consistently 5% to 8% of DNA is not extracted by DNase under these conditions. The average DNA values thus obtained for diploid and tetraploid nuclei of adult rat liver are 7.7 and 15.6 pg (10-12 g), respectively. Definite advantages of this procedure are: i) comparisons with biochemical determinations to give DNA values for each class of ploidy, ii) comparisons with histophotometry of the Feulgen dye-DNA complex to give absolute values instead of arbitrary units. PMID:5792332

  9. Quantitative determination of fluoride in pure water using luminescent europium complexes.

    PubMed

    Butler, Stephen J

    2015-07-11

    Two luminescent probes [Eu.L¹⁻²]⁺ are reported for the rapid detection of fluoride in water. Probes [Eu.L¹⁻²]⁺ exhibit exceptional enhancements in Eu emission in the presence of fluoride, permitting its selective determination within the environmentally relevant concentration range (20-210 μM).

  10. Quantitative determination of chitosan and the percentage of free amino groups.

    PubMed

    Curotto, E; Aros, F

    1993-06-01

    Chitosans were quantified with ninhydrin, a reagent normally used for recognizing and quantifying amino groups. The reaction was time dependent and there was no effect when different acids were added to the mixture. This method was used to determine the percentage of free amino groups in chitosans of different origins. PMID:8317699

  11. Quantitative Ultrasound-Assisted Extraction for Trace-Metal Determination: An Experiment for Analytical Chemistry

    ERIC Educational Resources Information Center

    Lavilla, Isela; Costas, Marta; Pena-Pereira, Francisco; Gil, Sandra; Bendicho, Carlos

    2011-01-01

    Ultrasound-assisted extraction (UAE) is introduced to upper-level analytical chemistry students as a simple strategy focused on sample preparation for trace-metal determination in biological tissues. Nickel extraction in seafood samples and quantification by electrothermal atomic absorption spectrometry (ETAAS) are carried out by a team of four…

  12. Quantitative determination of chitosan and the percentage of free amino groups.

    PubMed

    Curotto, E; Aros, F

    1993-06-01

    Chitosans were quantified with ninhydrin, a reagent normally used for recognizing and quantifying amino groups. The reaction was time dependent and there was no effect when different acids were added to the mixture. This method was used to determine the percentage of free amino groups in chitosans of different origins.

  13. Quantitative Determination of Levonorgestrel in Fish Plasma using UPLC-MS/MS

    EPA Science Inventory

    In this study, a sensitive high-performance liquid chromatography electrospray tandem mass spectrometric method was developed for the determination of levonorgestrel in fish plasma using levonorgestrel-d6 as an internal standard (IS). In the laboratory, the fish cunner, (Tautogol...

  14. Uranium isotopes quantitatively determined by modified method of atomic absorption spectrophotometry

    NASA Technical Reports Server (NTRS)

    Lee, G. H.

    1967-01-01

    Hollow-cathode discharge tubes determine the quantities of uranium isotopes in a sample by using atomic absorption spectrophotometry. Dissociation of the uranium atoms allows a large number of ground state atoms to be produced, absorbing the incident radiation that is different for the two major isotopes.

  15. A fast semi-quantitative method for Plutonium determination in an alpine firn/ice core

    NASA Astrophysics Data System (ADS)

    Gabrieli, J.; Cozzi, G.; Vallelonga, P.; Schwikowski, M.; Sigl, M.; Boutron, C.; Barbante, C.

    2009-04-01

    deposition decreased very sharply reaching a minimum in 1967. The third period (1967-1975) is characterized by irregular Pu profiles with smaller peaks (about 20-30% compared to the 1964 peak) which could be due to French and Chinese tests. Comparison with the Pu profiles obtained from the Col du Dome and Belukha ice cores by AMS (Accelerator Mass Spectrometry) shows very good agreement. Considering the semi-quantitative method and the analytical uncertainty, the results are also quantitatively comparable. However, the Pu concentrations at Colle Gnifetti are normally 2-3 times greater than in Col du Dome. This could be explained by different air mass transport or, more likely, different accumulation rates at each site.

  16. Determination of Quantitative Trait Loci (QTL) for Early Maturation in Rainbow Trout (Oncorhynchus mykiss)

    PubMed Central

    Haidle, Lisa; Janssen, Jennifer E.; Gharbi, Karim; Moghadam, Hooman K.; Ferguson, Moira M.

    2008-01-01

    To identify quantitative trait loci (QTL) influencing early maturation (EM) in rainbow trout (Oncorhynchus mykiss), a genome scan was performed using 100 microsatellite loci across 29 linkage groups. Six inter-strain paternal half-sib families using three inter-strain F1 brothers (approximately 50 progeny in each family) derived from two strains that differ in the propensity for EM were used in the study. Alleles derived from both parental sources were observed to contribute to the expression of EM in the progeny of the brothers. Four genome-wide significant QTL regions (i.e., RT-8, -17, -24, and -30) were observed. EM QTL detected on RT-8 and -24 demonstrated significant and suggestive QTL effects in both male and female progeny. Furthermore, within both male and female full-sib groupings, QTL on RT-8 and -24 were detected in two or more of the five parents used. Significant genome-wide and several strong chromosome-wide QTL for EM localized to different regions in males and females, suggesting some sex-specific control. Namely, QTL detected on RT-13, -15, -21, and -30 were associated with EM only in females, and those on RT-3, -17, and -19 were associated with EM only in males. Within the QTL regions identified, a comparison of syntenic EST markers from the rainbow trout linkage map with the zebrafish (Danio rerio) genome identified several putative candidate genes that may influence EM. Electronic supplementary material The online version of this article (doi:10.1007/s10126-008-9098-5) contains supplementary material, which is available to authorized users. PMID:18491191

  17. A convenient method for the quantitative determination of elemental sulfur in coal by HPLC analysis of perchloroethylene extracts

    USGS Publications Warehouse

    Buchanan, D.H.; Coombs, K.J.; Murphy, P.M.; Chaven, C.

    1993-01-01

    A convenient method for the quantitative determination of elemental sulfur in coal is described. Elemental sulfur is extracted from the coal with hot perchloroethylene (PCE) (tetrachloroethene, C2Cl4) and quantitatively determined by HPLC analysis on a C18 reverse-phase column using UV detection. Calibration solutions were prepared from sublimed sulfur. Results of quantitative HPLC analyses agreed with those of a chemical/spectroscopic analysis. The HPLC method was found to be linear over the concentration range of 6 ?? 10-4 to 2 ?? 10-2 g/L. The lower detection limit was 4 ?? 10-4 g/L, which for a coal sample of 20 g is equivalent to 0.0006% by weight of coal. Since elemental sulfur is known to react slowly with hydrocarbons at the temperature of boiling PCE, standard solutions of sulfur in PCE were heated with coals from the Argonne Premium Coal Sample program. Pseudo-first-order uptake of sulfur by the coals was observed over several weeks of heating. For the Illinois No. 6 premium coal, the rate constant for sulfur uptake was 9.7 ?? 10-7 s-1, too small for retrograde reactions between solubilized sulfur and coal to cause a significant loss in elemental sulfur isolated during the analytical extraction. No elemental sulfur was produced when the following pure compounds were heated to reflux in PCE for up to 1 week: benzyl sulfide, octyl sulfide, thiane, thiophene, benzothiophene, dibenzothiophene, sulfuric acid, or ferrous sulfate. A sluury of mineral pyrite in PCE contained elemental sulfur which increased in concentration with heating time. ?? 1993 American Chemical Society.

  18. Ultratrace Level Determination and Quantitative Analysis of Kidney Injury Biomarkers in Patient Samples Attained by Zinc Oxide Nanorods

    PubMed Central

    Singh, Manpreet; Alabanza, Anginelle; Gonzalez, Lorelis E.; Wang, Weiwei; Reeves, W. Brian; Hahm, Jong-in

    2016-01-01

    Determining ultratrace amounts of protein biomarkers in patient samples in a straightforward and quantitative manner is extremely important for early disease diagnosis and treatment. Here, we successfully demonstrate the novel use of zinc oxide nanorods (ZnO NRs) in the ultrasensitive and quantitative detection of two acute kidney injury (AKI)-related protein biomarkers, tumor necrosis factor (TNF)-α and interleukin (IL)-8, directly from patient samples. We first validate the ZnO NRs-based IL-8 results via comparison with those obtained from using a conventional enzyme-linked immunosorbent method in samples from 38 individuals. We further assess the full detection capability of the ZnO NRs-based technique by quantifying TNF-α, whose levels in human urine are often below the detection limits of conventional methods. Using the ZnO NR platforms, we determine the TNF-α concentrations of all 46 patient samples tested, down to the fg/mL level. Subsequently, we screen for TNF-α levels in approximately 50 additional samples collected from different patient groups in order to demonstrate a potential use of the ZnO NRs-based assay in assessing cytokine levels useful for further clinical monitoring. Our research efforts demonstrate that ZnO NRs can be straightforwardly employed in the rapid, ultrasensitive, quantitative, and simultaneous detection of multiple AKI-related biomarkers directly in patient urine samples, providing an unparalleled detection capability beyond those of conventional analysis methods. Additional key advantages of the ZnO NRs-based approach include a fast detection speed, low-volume assay condition, multiplexing ability, and easy automation/integration capability to existing fluorescence instrumentation. Therefore, we anticipate that our ZnO NRs-based detection method will be highly beneficial for overcoming the frequent challenges in early biomarker development and treatment assessment, pertaining to the facile and ultrasensitive quantification of

  19. Ultratrace level determination and quantitative analysis of kidney injury biomarkers in patient samples attained by zinc oxide nanorods.

    PubMed

    Singh, Manpreet; Alabanza, Anginelle; Gonzalez, Lorelis E; Wang, Weiwei; Reeves, W Brian; Hahm, Jong-in

    2016-02-28

    Determining ultratrace amounts of protein biomarkers in patient samples in a straightforward and quantitative manner is extremely important for early disease diagnosis and treatment. Here, we successfully demonstrate the novel use of zinc oxide nanorods (ZnO NRs) in the ultrasensitive and quantitative detection of two acute kidney injury (AKI)-related protein biomarkers, tumor necrosis factor (TNF)-α and interleukin (IL)-8, directly from patient samples. We first validate the ZnO NRs-based IL-8 results via comparison with those obtained from using a conventional enzyme-linked immunosorbent method in samples from 38 individuals. We further assess the full detection capability of the ZnO NRs-based technique by quantifying TNF-α, whose levels in human urine are often below the detection limits of conventional methods. Using the ZnO NR platforms, we determine the TNF-α concentrations of all 46 patient samples tested, down to the fg per mL level. Subsequently, we screen for TNF-α levels in approximately 50 additional samples collected from different patient groups in order to demonstrate a potential use of the ZnO NRs-based assay in assessing cytokine levels useful for further clinical monitoring. Our research efforts demonstrate that ZnO NRs can be straightforwardly employed in the rapid, ultrasensitive, quantitative, and simultaneous detection of multiple AKI-related biomarkers directly in patient urine samples, providing an unparalleled detection capability beyond those of conventional analysis methods. Additional key advantages of the ZnO NRs-based approach include a fast detection speed, low-volume assay condition, multiplexing ability, and easy automation/integration capability to existing fluorescence instrumentation. Therefore, we anticipate that our ZnO NRs-based detection method will be highly beneficial for overcoming the frequent challenges in early biomarker development and treatment assessment, pertaining to the facile and ultrasensitive quantification

  20. Capillary zone electrophoresis for separation and quantitative determination of mexiletine and its main phase I metabolites.

    PubMed

    Bruno, Claudio; Cavalluzzi, Maria Maddalena; Carocci, Alessia; Catalano, Alessia; Franchini, Carlo; Lentini, Giovanni

    2013-03-01

    The simultaneous separation and quantification of the analytes within the minimum analysis time and the maximum resolution and efficiency are the main objectives in the development of a capillary electrophoretic method for the determination of solutes. In this paper we describe a specific, sensitive and robust method, using capillary zone electrophoresis with internal standard and UV detection, for the separation and quantification of the anti-arrhythmic drug mexiletine, its main phase I metabolites, and its main nitrogenous degradation product. PMID:23826880