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Sample records for acid sample preparation

  1. Nucleic Acid Sample Preparation using Spontaneous Biphasic Plug Flow

    PubMed Central

    Thomas, Peter C.; Strotman, Lindsay N.; Theberge, Ashleigh B.; Berthier, Erwin; O’Connell, Rachel; Loeb, Jennifer M.; Berry, Scott M.; Beebe, David J.

    2013-01-01

    Nucleic acid (NA) extraction and purification has become a common technique in both research and clinical laboratories. Current methods require repetitive wash steps with a pipette that is laborious and time consuming making it inefficient for clinical settings. We present here a simple technique that relies on spontaneous biphasic plug flow inside a capillary to achieve sample preparation. By filling the sample with oil, aqueous contaminants were displaced from the captured NA without pipetting wash buffers or use of external force and equipment. mRNA from mammalian cell culture was purified and PCR amplification showed similar threshold cycle values as those obtained from a commercially available kit. HIV viral like particles were spiked into serum and a 5-fold increase in viral RNA extraction yield was achieved compared to the conventional wash method. In addition, viral RNA was successfully purified from human whole blood, and a limit of detection of approximately 14 copies of RNA extracted per sample. The results demonstrate the utility of the current technique for nucleic acid purification for clinical purposes, and the overall approach provides a potential method to implement nucleic acid testing in low resource settings. PMID:23941230

  2. Nucleic acid sample preparation using spontaneous biphasic plug flow.

    PubMed

    Thomas, Peter C; Strotman, Lindsay N; Theberge, Ashleigh B; Berthier, Erwin; O'Connell, Rachel; Loeb, Jennifer M; Berry, Scott M; Beebe, David J

    2013-09-17

    Nucleic acid (NA) extraction and purification has become a common technique in both research and clinical laboratories. Current methods require repetitive wash steps with a pipet that are laborious and time-consuming, making the procedure inefficient for clinical settings. We present here a simple technique that relies on spontaneous biphasic plug flow inside a capillary to achieve sample preparation. By filling the sample with oil, aqueous contaminants were displaced from the captured NA without pipetting wash buffers or use of external force and equipment. mRNA from mammalian cell culture was purified, and polymerase chain reaction (PCR) amplification showed similar threshold cycle values as those obtained from a commercially available kit. Human immunodeficiency virus (HIV) viral-like particles were spiked into serum and a 5-fold increase in viral RNA extraction yield was achieved compared to the conventional wash method. In addition, viral RNA was successfully purified from human whole blood, and a limit of detection of approximately 14 copies of RNA extracted per sample was determined. The results demonstrate the utility of the current technique for nucleic acid purification for clinical purposes, and the overall approach provides a potential method to implement nucleic acid testing in low-resource settings.

  3. Digital microfluidic hub for automated nucleic acid sample preparation.

    SciTech Connect

    He, Jim; Bartsch, Michael S.; Patel, Kamlesh D.; Kittlaus, Eric A.; Remillared, Erin M.; Pezzola, Genevieve L.; Renzi, Ronald F.; Kim, Hanyoup

    2010-07-01

    We have designed, fabricated, and characterized a digital microfluidic (DMF) platform to function as a central hub for interfacing multiple lab-on-a-chip sample processing modules towards automating the preparation of clinically-derived DNA samples for ultrahigh throughput sequencing (UHTS). The platform enables plug-and-play installation of a two-plate DMF device with consistent spacing, offers flexible connectivity for transferring samples between modules, and uses an intuitive programmable interface to control droplet/electrode actuations. Additionally, the hub platform uses transparent indium-tin oxide (ITO) electrodes to allow complete top and bottom optical access to the droplets on the DMF array, providing additional flexibility for various detection schemes.

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

    SciTech Connect

    Cary, Robert E.

    2015-12-08

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

  5. Universal nucleic acids sample preparation method for cells, spores and their mixture

    DOEpatents

    Bavykin, Sergei [Darien, IL

    2011-01-18

    The present invention relates to a method for extracting nucleic acids from biological samples. More specifically the invention relates to a universal method for extracting nucleic acids from unidentified biological samples. An advantage of the presently invented method is its ability to effectively and efficiently extract nucleic acids from a variety of different cell types including but not limited to prokaryotic or eukaryotic cells and/or recalcitrant organisms (i.e. spores). Unlike prior art methods which are focused on extracting nucleic acids from vegetative cell or spores, the present invention effectively extracts nucleic acids from spores, multiple cell types or mixtures thereof using a single method. Important that the invented method has demonstrated an ability to extract nucleic acids from spores and vegetative bacterial cells with similar levels effectiveness. The invented method employs a multi-step protocol which erodes the cell structure of the biological sample, isolates, labels, fragments nucleic acids and purifies labeled samples from the excess of dye.

  6. Study of sample preparation for quantitative analysis of amino acids in human sweat by liquid chromatography-tandem mass spectrometry.

    PubMed

    Delgado-Povedano, M M; Calderón-Santiago, M; Priego-Capote, F; Luque de Castro, M D

    2016-01-01

    The determination of physiological levels of amino acids is important to aid in the diagnosis and treatment of several diseases and nutritional status of individuals. Amino acids are frequently determined in biofluids such as blood (serum or plasma) and urine; however, there are less common biofluids with different concentration profiles of amino acids that could be of interest. One of these biofluids is sweat that can be obtained in a non-invasive manner and is characterized by low complex composition. The analysis of amino acids in human sweat requires the development of sample preparation strategies according to the sample matrix and small collected volume. The influence of sample preparation on the quantitative analysis of amino acids in sweat by LC-MS/MS has been assessed through a comparison between two strategies: dilution of sweat and centrifugal microsolid-phase extraction (c-μSPE). In both cases, several dilution factors were assayed for in-depth knowledge of the matrix effects, and the use of c-μSPE provided the best results in terms of accuracy. The behavior of the target analytes was a function of the dilution factor, thus providing a pattern for sample preparation that depended on the amino acid to be determined. The concentration of amino acids in sweat ranges between 6.20 ng mL(-1) (for homocysteine) and 259.77 µg mL(-1) (for serine) with precision, expressed as relative standard deviation, within 1.1-21.4%.

  7. Final Report Nucleic Acid System - PCR, Multiplex Assays and Sample Preparation Project

    SciTech Connect

    Koopman, R.P.; Langlois, R.G.; Nasarabadi, S.; Benett, W.J.; Richards, J.B.; Hadley, D.R.; Miles, R.R.; Brown, S.B.; Stratton, P.L.; Milanovich, F.P.

    2001-04-20

    The objective of this project was to reduce to practice the detection and identification of biological warfare pathogens by the nucleic acid recognition technique of PCR (polymerase chain reaction). This entailed not only building operationally functional instrumentation but also developing the chemical assays for detection of priority pathogens. This project had two principal deliverables: (1) design, construct, test and deliver a 24 chamber, multiplex capable suitcase sized PCR instrument, and (2) develop and reduce to practice a multiplex assay for the detection of PCR product by flow cytometry. In addition, significant resources were allocated to test and evaluation of the Hand-held Advanced Nucleic Acid Analyzer (HANAA). This project helps provide the signature and intelligence gathering community the ability to perform, on-site or remote, rapid analysis of environmental or like samples for the presence of a suite of biological warfare pathogens.

  8. Strategies for automated sample preparation, nucleic acid purification, and concentration of low-target-number nucleic acids in environmental and food processing samples

    NASA Astrophysics Data System (ADS)

    Bruckner-Lea, Cynthia J.; Holman, David A.; Schuck, Beatrice L.; Brockman, Fred J.; Chandler, Darrell P.

    1999-01-01

    The purpose of this work is to develop a rapid, automated system for nucleic acid purification and concentration from environmental and food processing samples. Our current approach involves off-line filtration and cell lysis (ballistic disintegration) functions in appropriate buffers followed by automated nucleic acid capture and purification on renewable affinity matrix microcolumns. Physical cell lysis and renewable affinity microcolumns eliminate the need for toxic organic solvents, enzyme digestions or other time- consuming sample manipulations. Within the renewable affinity microcolumn, we have examined nucleic acid capture and purification efficiency with various microbead matrices (glass, polymer, paramagnetic), surface derivitization (sequence-specific capture oligonucleotides or peptide nucleic acids), and DNA target size and concentration under variable solution conditions and temperatures. Results will be presented comparing automated system performance relative to benchtop procedures for both clean (pure DNA from a laboratory culture) and environmental (soil extract) samples, including results which demonstrate 8 minute purification and elution of low-copy nucleic acid targets from a crude soil extract in a form suitable for PCR or microarray-based detectors. Future research will involve the development of improved affinity reagents and complete system integration, including upstream cell concentration and cell lysis functions and downstream, gene-based detectors. Results of this research will ultimately lead to improved processes and instrumentation for on-line, automated monitors for pathogenic micro-organisms in food, water, air, and soil samples.

  9. Modular microfluidic system for biological sample preparation

    DOEpatents

    Rose, Klint A.; Mariella, Jr., Raymond P.; Bailey, Christopher G.; Ness, Kevin Dean

    2015-09-29

    A reconfigurable modular microfluidic system for preparation of a biological sample including a series of reconfigurable modules for automated sample preparation adapted to selectively include a) a microfluidic acoustic focusing filter module, b) a dielectrophoresis bacteria filter module, c) a dielectrophoresis virus filter module, d) an isotachophoresis nucleic acid filter module, e) a lyses module, and f) an isotachophoresis-based nucleic acid filter.

  10. Microscopic and mesoscopic structural features of an activated carbon sample, prepared from sorghum via activation by phosphoric acid

    SciTech Connect

    Temleitner, László; Pusztai, László; Rubio-Arroyo, Manuel F.; Aguilar-López, Sergio; Pizio, Orest

    2012-12-15

    Graphical abstract: Display Omitted Highlights: ► Preparation of a new activated carbon sample from sorghum. ► Characterization by adsorption/desorption methods. ► Determination of the structure by synchrotron X-ray diffraction. ► The sample is amorphous and contains distorted graphene fragments. ► A characteristic nanoscale distance is established from the radial distribution function. -- Abstract: An acidic chemical activation procedure has been used for preparing activated carbon with a surface area exceeding 1000 m{sup 2}/g from sorghum. In order to reveal structural features, synchrotron X-ray diffraction measurements have been performed. The structure of the material has been characterized by the total scattering structure factor and the radial distribution function describing short-range arrangement of atoms at distances of the order of a few atomic diameters as well as correlations at a longer scale, of the order of nanometers. The atomic arrangement has been found to be consistent with that of amorphous graphite-like carbon. As far as the mesoscopic structure is concerned, the presence of a characteristic distance is suggested on the basis of the clear nanometer scale oscillations of the radial distribution function, which distance may be assigned as the mesopore size in the material. It is suggested that the approach devized here may later be applied routinely for other activated carbon samples, too, for characterizing atomic and nanoscale order simultaneously.

  11. Rapid and automated sample preparation for nucleic acid extraction on a microfluidic CD (compact disk)

    NASA Astrophysics Data System (ADS)

    Kim, Jitae; Kido, Horacio; Zoval, Jim V.; Gagné, Dominic; Peytavi, Régis; Picard, François J.; Bastien, Martine; Boissinot, Maurice; Bergeron, Michel G.; Madou, Marc J.

    2006-01-01

    Rapid and automated preparation of PCR (polymerase chain reaction)-ready genomic DNA was demonstrated on a multiplexed CD (compact disk) platform by using hard-to-lyse bacterial spores. Cell disruption is carried out while beadcell suspensions are pushed back and forth in center-tapered lysing chambers by angular oscillation of the disk - keystone effect. During this lysis period, the cell suspensions are securely held within the lysing chambers by heatactivated wax valves. Upon application of a remote heat to the disk in motion, the wax valves release lysate solutions into centrifuge chambers where cell debris are separated by an elevated rotation of the disk. Only debris-free DNA extract is then transferred to collection chambers by capillary-assisted siphon and collected for heating that inactivates PCR inhibitors. Lysing capacity was evaluated using a real-time PCR assay to monitor the efficiency of Bacillus globigii spore lysis. PCR analysis showed that 5 minutes' CD lysis run gave spore lysis efficiency similar to that obtained with a popular commercial DNA extraction kit (i.e., IDI-lysis kit from GeneOhm Sciences Inc.) which is highly efficient for microbial cell and spore lysis. This work will contribute to the development of an integrated CD-based assay for rapid diagnosis of infectious diseases.

  12. Microfluidic Sample Preparation for Immunoassays

    SciTech Connect

    Visuri, S; Benett, W; Bettencourt, K; Chang, J; Fisher, K; Hamilton, J; Krulevitch, P; Park, C; Stockton, C; Tarte, L; Wang, A; Wilson, T

    2001-08-09

    Researchers at Lawrence Livermore National Laboratory are developing means to collect and identify fluid-based biological pathogens in the forms of proteins, viruses, and bacteria. to support detection instruments, they are developing a flexible fluidic sample preparation unit. The overall goal of this Microfluidic Module is to input a fluid sample, containing background particulates and potentially target compounds, and deliver a processed sample for detection. They are developing techniques for sample purification, mixing, and filtration that would be useful to many applications including immunologic and nucleic acid assays. Many of these fluidic functions are accomplished with acoustic radiation pressure or dielectrophoresis. They are integrating these technologies into packaged systems with pumps and valves to control fluid flow through the fluidic circuit.

  13. Sample preparation techniques.

    PubMed

    Baumgartner, W A; Hill, V A

    1993-12-01

    Evidentiary false positives are caused by passive exposure to drugs in the environment rather than by active use of drugs. The avoidance of such positives is essential for both hair and urine analysis. Hair analysis enjoys the advantage over urinalysis in having a number of approaches for making this distinction. These include: methylene blue staining of the hair specimen for selecting the appropriate wash solvent; application of hair digestion techniques for the complete release of chemically unaltered analytes; the determination of three diagnostic ratios from wash and digestion data; the measurement of metabolite:drug ratios; the use of cut-off levels setting the limits for passive endogenous drug exposure; reproducibility of results (including segmental analysis) with a newly collected hair specimen; and the reporting of results as either negative, positive, or contaminated. Our sample preparation procedures have been effectively applied to the analyses of nearly 200,000 specimens, i.e. to approximately one million drug analyses for cocaine, opiates, methamphetamine, phencyclidine or marijuana. On the basis of this experience we conclude that hair analysis is a safe and effective method for workplace drug testing.

  14. Microwave sample preparation for analysis of metals in environmental samples

    SciTech Connect

    Collins, L.W.

    1996-10-01

    The unique nature of microwave energy enhances heating efficiency and improves acid digestion sample preparation. Faster sample preparation and improved precision of the analysis occur. These results will be illustrated in this presentation using various standard reference materials and environmentally important samples. The analytical microwave system used offers accurate temperature and pressure feedback control through the use of a hand-held controller or PC-based control. Digestions are performed in patented, user-friendly microwave vessels. USEPA Method 3015, {open_quotes}Microwave-Assisted Acid Digestion of Aqueous Samples and Extracts,{close_quotes} is properly performed when the sample is heated to 170{degrees}C within 10 minutes, and maintained for an additional 10 minutes. USEPA Method 3051, {open_quotes}Microwave-Assisted Acid Digestion of Sediments, Sludges, Soils, and Oils,{close_quotes} is properly performed when the sample is heated to 175{degrees}C within 5.5 minutes, and maintained at 175{degrees}C for an additional 4.5 minutes. After the timesaving microwave digestion period, the samples were analyzed for metals by ICP-AES. Excellent accuracy and precision were obtained, in addition to 90% time reduction when using microwave sample preparation.

  15. Universal sample preparation method integrating trichloroacetic acid/acetone precipitation with phenol extraction for crop proteomic analysis.

    PubMed

    Wu, Xiaolin; Xiong, Erhui; Wang, Wei; Scali, Monica; Cresti, Mauro

    2014-02-01

    Crop plants contain large amounts of secondary compounds that interfere with protein extraction and gel-based proteomic analysis. Thus, a protein extraction protocol that can be easily applied to various crop materials with minimal optimization is essential. Here we describe a universal protocol for total protein extraction involving trichloroacetic acid (TCA)/acetone precipitation followed by SDS and phenol extraction. Through SDS extraction, the proteins precipitated by the TCA/acetone treatment can be fully resolubilized and then further purified by phenol extraction. This protocol combines TCA/acetone precipitation, which aggressively removes nonprotein compounds, and phenol extraction, which selectively dissolves proteins, resulting in effective purification of proteins from crop tissues. This protocol can also produce high-quality protein preparations from various recalcitrant tissues, and therefore it has a wide range of applications in crop proteomic analysis. Designed to run on a small scale, this protocol can be completed within 5 h.

  16. Routine sample preparation and HPLC analysis for ascorbic acid (vitamin C) determination in wheat plants and Arabidopsis leaf tissues.

    PubMed

    Szalai, Gabriella; Janda, T; Pál, Magda

    2014-06-01

    Plants have developed various mechanisms to protect themselves against oxidative stress. One of the most important non-enzymatic antioxidants is ascorbic acid. There is thus a need for a rapid, sensitive method for the analysis of the reduced and oxidised forms of ascorbic acid in crop plants. In this paper a simple, economic, selective, precise and stable HPLC method is presented for the detection of ascorbate in plant tissue. The sensitivity, the short retention time and the simple isocratic elution mean that the method is suitable for the routine quantification of ascorbate in a high daily sample number. The method has been found to be better than previously reported methods, because of the use of an economical, readily available mobile phase, UV detection and the lack of complicated extraction procedures. The method has been tested on Arabidopsis plants with different ascorbate levels and on wheat plants during Cd stress.

  17. Suppression of lysophosphatidic acid and lysophosphatidylcholine formation in the plasma in vitro: proposal of a plasma sample preparation method for laboratory testing of these lipids.

    PubMed

    Nakamura, Kazuhiro; Kishimoto, Tatsuya; Ohkawa, Ryunosuke; Okubo, Shigeo; Tozuka, Minoru; Yokota, Hiromitsu; Ikeda, Hitoshi; Ohshima, Noriko; Mizuno, Koji; Yatomi, Yutaka

    2007-08-01

    It is now established that lysophosphatidic acid (LPA) and lysophosphatidylcholine (LPC) play important roles in a variety of biological responses, especially in the area of vascular biology, and determination of their concentrations in the plasma is believed to be clinically relevant. Preparation of the measurement samples is a difficult task, however, because the blood levels of these lipids can be easily increased by in vitro manipulation after venepuncture. In this study, we examined the optimal conditions for the preparation of plasma samples for the measurement of LPA and LPC. It appears that regulation of platelet activation and the enzymatic activity of lysophospholipase D/autotaxin and lecithin-cholesterol acyltransferase is important to suppress the undesirable formation of LPA and LPC after venepuncture. We found that in vitro formation of LPA and LPC was negligible when whole blood samples were mixed with 7.5 mM EDTA plus 10% (v/v) citrate-theophylline-adenosine-dipyridamole (CTAD) and when all of the procedures, including the plasma preparation and preservation until measurement, were performed at 4 degrees C. Thus, although the plasma levels of LPA and LPC can be easily altered, laboratory testing of these important bioactive lipids for clinical purposes may be conducted reliably if the samples are prepared under stringent conditions.

  18. Statistical evaluation of fatty acid profile and cholesterol content in fish (common carp) lipids obtained by different sample preparation procedures.

    PubMed

    Spiric, Aurelija; Trbovic, Dejana; Vranic, Danijela; Djinovic, Jasna; Petronijevic, Radivoj; Matekalo-Sverak, Vesna

    2010-07-05

    the second principal component (PC2) is recorded by C18:3 n-3, and C20:3 n-6, being present in a higher amount in the samples treated by the modified Soxhlet extraction, while C22:5 n-3, C20:3 n-3, C22:1 and C20:4, C16 and C18 negatively influence the score values of the PC2, showing significantly increased level in the samples treated by ASE method. Hotelling's paired T-square test used on the first three principal components for confirmation of differences in individual fatty acid content obtained by ASE and Soxhlet method in carp muscle showed statistically significant difference between these two data sets (T(2)=161.308, p<0.001).

  19. Sample preparation for STED microscopy.

    PubMed

    Wurm, Christian A; Neumann, Daniel; Schmidt, Roman; Egner, Alexander; Jakobs, Stefan

    2010-01-01

    Since the discovery of the diffraction barrier in the late nineteenth century, it has been commonly accepted that with far-field optical microscopy it is not possible to resolve structural details considerably finer than half the wavelength of light. The emergence of STED microscopy showed that, at least for fluorescence imaging, these limits can be overcome. Since STED microscopy is a far-field technique, in principle, the same sample preparation as for conventional confocal microscopy may be utilized. The increased resolution, however, requires additional precautions to ensure the structural preservation of the specimen. We present robust protocols to generate test samples for STED microscopy. These protocols for bead samples and immunolabeled mammalian cells may be used as starting points to adapt existing labeling strategies for the requirements of sub-diffraction resolution microscopy.

  20. An improved sample preparation for an LC method used in the age estimation based on aspartic acid racemization from human dentin.

    PubMed

    Yekkala, Raja; Meers, Carine; Hoogmartens, Jos; Lambrichts, Ivo; Willems, Guy; Van Schepdael, Ann

    2007-01-01

    The determination of age on the basis of aspartic acid (Asp) racemization in teeth is one of the most reliable and accurate methods to date. In this paper, the usefulness of HPLC coupled with fluorescence detection for determination of Asp racemization was evaluated. A modified sample preparation is proposed for better stability of o-phthaldialdehyde-N-acetyl-L-cysteine derivatives of D/L-Asp (due to the instability below pH 7). To ensure the accuracy of the method, the validation parameters' specificity, precision, linearity, and LOD were determined. Three dentin samples of premolar teeth, extracted from living individuals (bucco-lingual longitudinal sections of 1 mm thickness), were analyzed and quantitative results are discussed.

  1. Development of sample preparation method for isoliquiritigenin, liquiritin, and glycyrrhizic acid analysis in licorice by ionic liquids-ultrasound based extraction and high-performance liquid chromatography detection.

    PubMed

    Yang, Lei; Li, Li-li; Liu, Ting-ting; Zu, Yuan-gang; Yang, Feng-jian; Zhao, Chun-jian; Zhang, Lin; Chen, Xiao-qiang; Zhang, Zhong-hua

    2013-05-01

    An ionic liquid-based ultrasonic-assisted extraction (ILUAE) method had been used for the effective extraction of isoliquiritigenin (IQ), liquiritin (LQ) and glycyrrhizic acid (GA) from licorice. The ionic liquids with different cations and anions were investigated in this work and 0.5 M 1-butyl-3-methylimidazolium bromide solution was selected as solvent. In addition, the technical parameters including soaking time, solid-liquid ratio, ultrasonic power and time were optimized. Compared with the conventional solvent extraction, the proposed approach exhibited higher efficiency, which indicated the ILUAE was an efficient, rapid and simple sample preparation technique. There was no degradation of the target analytes had been observed at the optimum conditions which was evidenced by the stability studies performed with standard of IQ, LQ and GA. The proposed method also showed high reproducibility and was environmental friendly.

  2. Comparative study of sample preparation techniques coupled to GC for the analysis of halogenated acetic acids (HAAs) acids in tap water.

    PubMed

    Waseem, Sadia; Abdullah, Md Pauzi

    2010-03-01

    Halogenated acetic acids (HAAs) are one of the most common disinfection by-products formed during chlorination of drinking water. Currently, there are three U.S. Environmental Protection Agency-approved methods for analysis of HAAs in drinking water: U.S. EPA method 552.2, Standard Method 6251, and U.S. EPA method 552.3. The current U.S.EPA-approved HAA analysis methods require tedious and time-consuming liquid-liquid extraction (LLE) and the use of hazardous chemicals. Besides U.S. EPA methods, capillary electrophoresis (CE), liquid chromatography (LC), including ion chromatography (IC), and electrospray ionization mass spectrometry (ESI-MS) have been applied in order to determine the HAAs in portable water with high detection limits. Detection limits required to analyze portable water samples can be regularly achieved only by gas chromatography-electron capture detector (ECD) and ESI-MS. In this study, improved gas chromatography-mass spectrometry (GC-MS) method was able to achieve HAAs analysis at low detection limits. Thus, a safe and rapid method is needed for the micro-determination of HAAs. A method involving solid-phase extraction (SPE) followed by GC-MS was developed to determine the HAAs in tap water. Selectivity, percent recovery, and detection limit studies were carried out on a LC-SAX (quaternary ammonium anion exchanger) SPE. Under optimized conditions, average recoveries for all nine HAAs spiked in drinking water samples ranged from 78.7% to 100%. The relative standard deviation data was found to range from 1.0% to 12.5% based upon five repeat recovery experiments, and estimated detection limit ranging between 0.16-0.009 microg/L was obtained. On this basis, SPE was studied as a possible alternative to LLE for the analysis of HAAs in water. Finally, the performance of the SPE-GC-MS with spiked drinking water samples was tested, and the results were compared with those obtained using LLE-GC-ECD. The method was applied for determination of HAAs in

  3. Automated Sample Preparation (ASP): Development of a Rapid Method to Sequentially Isolate Nucleic Acids and Protein from Any Sample Type by a Cartridge-Based System

    DTIC Science & Technology

    2013-11-27

    goals. Specifically, CUBRC will design and manufacture a prototype cartridge(s) and test the prototype cartridge for its ability to isolate each...analyte individually and in succession. Testing will be performed on both laboratory derived samples 1. REPORT DATE (DD-MM-YYYY) 4. TITLE AND SUBTITLE...CUBRC will design and manufacture a prototype cartridge(s) and test the prototype cartridge for its ability to isolate each analyte individually and in

  4. Construction of a portable sample preparation device with a magnetic poly(methacrylic acid-co-ethylene dimethacrylate) monolith as the extraction medium and its application in the enrichment of UV filters in water samples.

    PubMed

    Li, Jing; Xu, Li; Yu, Qiong-Wei; Shi, Zhi-Guo; Zhang, Ting; Liu, Yan

    2014-10-01

    A portable sample preparation device with a magnetic polymer monolith as the extraction medium was constructed. The monolith was synthesized by polymerizing methacrylic acid and ethylene dimethacrylate around a cylindrical magnet. In this way, the monolith with a magnetic core could be readily attached to the extraction device by magnetism. The constructed device was evaluated for the enrichment of UV filters in water samples, followed by high-performance liquid chromatographic analysis. The extraction efficiency for the targets was satisfactory with no matrix interference. Good linearities were obtained for the UV filters with the correlation coefficients >0.9986. The limits of detection and quantification for the UV filters were 0.3-0.8 and 1.0-2.4 ng/mL, respectively. The recoveries of the UV filters from the spiked water samples at the concentration of 100 ng/mL were 95.3-101.7%, with relative standard deviations <10%. Accordingly, the proposed portable device was demonstrated to be suitable for on-site simultaneous sampling, purification, and preconcentration within a single step.

  5. Microfluidic Sample Preparation for Medical Diagnostics.

    PubMed

    Cui, Francis; Rhee, Minsoung; Singh, Anup; Tripathi, Anubhav

    2015-01-01

    Fast and reliable diagnoses are invaluable in clinical care. Samples (e.g., blood, urine, and saliva) are collected and analyzed for various biomarkers to quickly and sensitively assess disease progression, monitor response to treatment, and determine a patient's prognosis. Processing conventional samples entails many manual time-consuming steps. Consequently, clinical specimens must be processed by skilled technicians before antigens or nucleic acids are detected, and these are often present at dilute concentrations. Recently, several automated microchip technologies have been developed that potentially offer many advantages over traditional bench-top extraction methods. The smaller length scales and more refined transport mechanisms that characterize these microfluidic devices enable faster and more efficient biomarker enrichment and extraction. Additionally, they can be designed to perform multiple tests or experimental steps on one integrated, automated platform. This review explores the current research on microfluidic methods of sample preparation that are designed to aid diagnosis, and covers a broad spectrum of extraction techniques and designs for various types of samples and analytes.

  6. Final Report BW Sample Collection& Preparation Device

    SciTech Connect

    Koopman, R P; Belgrader, P; Meyer, G; Benett, W J; Richards, J B; Hadley, D R; Stratton, P L; Milanovich, F P

    2002-01-31

    The objective of this project was to develop the technique needed to prepare a field collected sample for laboratory analysis and build a portable integrated biological detection instrument with new miniaturized and automated sample purification capabilities. The device will prepare bacterial spores, bacterial vegetative cells, and viral particles for PCR amplification.

  7. 40 CFR 761.323 - Sample preparation.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 761.323 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) TOXIC SUBSTANCES CONTROL... Remediation Waste Samples § 761.323 Sample preparation. (a) The comparison study requires analysis of a minimum of 10 samples weighing at least 300 grams each. Samples of PCB remediation waste used in...

  8. Innovative methods for inorganic sample preparation

    SciTech Connect

    Essling, A.M.; Huff, E.A.; Graczyk, D.G.

    1992-04-01

    Procedures and guidelines are given for the dissolution of a variety of selected materials using fusion, microwave, and Parr bomb techniques. These materials include germanium glass, corium-concrete mixtures, and zeolites. Emphasis is placed on sample-preparation approaches that produce a single master solution suitable for complete multielement characterization of the sample. In addition, data are presented on the soil microwave digestion method approved by the Environmental Protection Agency (EPA). Advantages and disadvantages of each sample-preparation technique are summarized.

  9. Microscale sample preparation for PCR of C. difficile infected stool

    PubMed Central

    Gillers, Sara; Atkinson, Christopher D.; Bartoo, Aaron C.; Mahalanabis, Madhumita; Boylan, Michael O.; Schwartz, John H.; Klapperich, Catherine; Singh, Satish K.

    2015-01-01

    In this paper, we describe the design of a microfluidic sample preparation chip for human stool samples infected with Clostridium difficile. We established a polymerase chain reaction able to distinguish C. difficile in the presence of several other organisms found in the normal intestinal flora. A protocol for on-chip extraction of nucleic acids from clinical samples is described that can detect target DNA down to 5.0×10−3 ng of template. The assay and sample preparation chip were then validated using known positive and known negative clinical samples. The work presented has potential applications in both the developed and developing world. PMID:19505511

  10. Sub-sampling and preparing forensic samples for pollen analysis.

    PubMed

    Horrocks, Mark

    2004-09-01

    The main forensic application of palynology is in providing associative evidence, assisting to prove or disprove a link between people and objects with places or with other people. Although identification and interpretation of pollen is a specialist job, sub-sampling and preparing pollen samples for analysis may be carried out by non-specialists. As few forensic laboratories have residing palynologists, laboratories may wish to reduce the cost of analysis or risk of contamination by doing their own sub-sampling and preparation. Presented is a practical guide for sub-sampling and preparing forensic samples for pollen analysis, providing a complete standard procedure for both the palynologist and non-specialist. Procedures for sub-sampling include a wide variety of materials commonly collected for forensic analysis (soil, clothing and other fabrics, footwear, twine and rope, firearms, granulated materials, plant and animal material, and illicit drugs), many of which palynologists will not be familiar with. Procedures for preparation of samples (pollen concentration) are presented as a detailed, step-by-step method. Minimizing the risks of laboratory and cross-sample contamination during sub-sampling and preparation is emphasized.

  11. Automated sample preparation for CE-SDS.

    PubMed

    Le, M Eleanor; Vizel, Alona; Hutterer, Katariina M

    2013-05-01

    Traditionally, CE with SDS (CE-SDS) places many restrictions on sample composition. Requirements include low salt content, known initial sample concentration, and a narrow window of final sample concentration. As these restrictions require buffer exchange for many sample types, sample preparation is often tedious and yields poor sample recoveries. To improve capacity and streamline sample preparation, an automated robotic platform was developed using the PhyNexus Micro-Extractor Automated Instrument (MEA) for both the reduced and nonreduced CE-SDS assays. This automated sample preparation normalizes sample concentration, removes salts and other contaminants, and adds the required CE-SDS reagents, essentially eliminating manual steps during sample preparation. Fc-fusion proteins and monoclonal antibodies were used in this work to demonstrate benefits of this approach when compared to the manual method. With optimized conditions, this application has demonstrated decreased analyst "hands on" time and reduced total assay time. Sample recovery greater than 90% can be achieved, regardless of initial composition and concentration of analyte.

  12. Soil sample preparation using microwave digestion for uranium analysis

    SciTech Connect

    MOHAGHEGHI,AMIR H.; PRESTON,ROSE; AKBARZADEH,MANSOOR; BAKHTIAR,STEVEN

    2000-04-05

    A new sample preparation procedure has been developed for digestion of soil samples for uranium analysis. The technique employs a microwave oven digestion system to digest the sample and to prepare it for separation chemistry and analysis. The method significantly reduces the volume of acids used, eliminates a large fraction of acid vapor emissions, and speeds up the analysis time. The samples are analyzed by four separate techniques: Gamma Spectrometry, Alpha Spectroscopy using the open digestion method, Kinetic Phosphorescence Analysis (KPA) using open digestion, and KPA by Microwave digestion technique. The results for various analytical methods are compared and used to confirm the validity of the new procedure. The details of the preparation technique along with its benefits are discussed.

  13. Molecularly imprinted polymers for bioanalytical sample preparation.

    PubMed

    Gama, Mariana Roberto; Bottoli, Carla Beatriz Grespan

    2017-02-01

    Molecularly imprinted polymers (MIP) are stable polymers with molecular recognition abilities, provided by the presence of a template during their synthesis, and are excellent materials with high selectivity for sample preparation in bioanalytical methods. This short review discusses aspects of MIP preparation and its applications as a sorbent material in pharmaceutical and biomedical analysis. MIP in different extraction configurations, including classical solid-phase extraction, solid-phase microextraction, magnetic molecularly imprinted solid-phase extraction, microextraction by packed sorbent and solid-phase extraction in pipette tips, are used to illustrate the good performance of this type of sorbent for sample preparation procedures of complex matrices, especially prior to bioanalytical approaches.

  14. [Recent advances in sample preparation methods of plant hormones].

    PubMed

    Wu, Qian; Wang, Lus; Wu, Dapeng; Duan, Chunfeng; Guan, Yafeng

    2014-04-01

    Plant hormones are a group of naturally occurring trace substances which play a crucial role in controlling the plant development, growth and environment response. With the development of the chromatography and mass spectroscopy technique, chromatographic analytical method has become a widely used way for plant hormone analysis. Among the steps of chromatographic analysis, sample preparation is undoubtedly the most vital one. Thus, a highly selective and efficient sample preparation method is critical for accurate identification and quantification of phytohormones. For the three major kinds of plant hormones including acidic plant hormones & basic plant hormones, brassinosteroids and plant polypeptides, the sample preparation methods are reviewed in sequence especially the recently developed methods. The review includes novel methods, devices, extractive materials and derivative reagents for sample preparation of phytohormones analysis. Especially, some related works of our group are included. At last, the future developments in this field are also prospected.

  15. Optimising uncertainty in physical sample preparation.

    PubMed

    Lyn, Jennifer A; Ramsey, Michael H; Damant, Andrew P; Wood, Roger

    2005-11-01

    Uncertainty associated with the result of a measurement can be dominated by the physical sample preparation stage of the measurement process. In view of this, the Optimised Uncertainty (OU) methodology has been further developed to allow the optimisation of the uncertainty from this source, in addition to that from the primary sampling and the subsequent chemical analysis. This new methodology for the optimisation of physical sample preparation uncertainty (u(prep), estimated as s(prep)) is applied for the first time, to a case study of myclobutanil in retail strawberries. An increase in expenditure (+7865%) on the preparatory process was advised in order to reduce the s(prep) by the 69% recommended. This reduction is desirable given the predicted overall saving, under optimised conditions, of 33,000 pounds Sterling per batch. This new methodology has been shown to provide guidance on the appropriate distribution of resources between the three principle stages of a measurement process, including physical sample preparation.

  16. Precise and automated microfluidic sample preparation.

    SciTech Connect

    Crocker, Robert W.; Patel, Kamlesh D.; Mosier, Bruce P.; Harnett, Cindy K.

    2004-07-01

    Autonomous bio-chemical agent detectors require sample preparation involving multiplex fluid control. We have developed a portable microfluidic pump array for metering sub-microliter volumes at flowrates of 1-100 {micro}L/min. Each pump is composed of an electrokinetic (EK) pump and high-voltage power supply with 15-Hz feedback from flow sensors. The combination of high pump fluid impedance and active control results in precise fluid metering with nanoliter accuracy. Automated sample preparation will be demonstrated by labeling proteins with fluorescamine and subsequent injection to a capillary gel electrophoresis (CGE) chip.

  17. Preparation and characterization of magnetic Wells-Dawson heteropoly acid nanoparticles for magnetic solid-phase extraction of aromatic amines in water samples.

    PubMed

    Amiri, Amirhassan; Saadati-Moshtaghin, Hamid Reza; Zonoz, Farokhzad Mohammadi; Targhoo, Azadeh

    2017-02-03

    In this work, aminopropyl modified silica-coated magnetite nanoparticles with Wells-Dawson heteropoly acid (P2W17Fe@APSCMNPs) was first synthesized and underwent highly efficient magnetic solid-phase extraction (MSPE) of aromatic amines from aqueous samples. The resulted nanomaterials were characterized with different physicochemical techniques such as Fourier transform infrared (FT-IR), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and vibrating sample magnetometry (VSM). Aniline, N,N-dimethylaniline, o-toluidine and 3-chloroaniline were selected as target compounds. The sample quantification was carried out using gas chromatography-flame ionization detector (GC-FID). Under optimal working conditions, the developed method showed good linearity (R>0.9912) in the range of 0.01-100ngmL(-1). The method displays detection limits (at an S/N ration of 3) in the range from 0.003 to 0.01ngmL(-1), and the limits of quantification (at an S/N ratio of 10) are between 0.01 and 0.04ngmL(-1). The enrichment factors (EFs) were in the range of 75-113. Relative standard deviations (RSDs) are 4.8-8.3%. The applicability of the developed method was examined by analyzing different water samples (river water, tap water, well water and wastewater) and the relative recovery values for the spiked water samples were found to be in the range of 90.7-99.8%.

  18. [Progress in sample preparation and analytical methods for trace polar small molecules in complex samples].

    PubMed

    Zhang, Qianchun; Luo, Xialin; Li, Gongke; Xiao, Xiaohua

    2015-09-01

    Small polar molecules such as nucleosides, amines, amino acids are important analytes in biological, food, environmental, and other fields. It is necessary to develop efficient sample preparation and sensitive analytical methods for rapid analysis of these polar small molecules in complex matrices. Some typical materials in sample preparation, including silica, polymer, carbon, boric acid and so on, are introduced in this paper. Meanwhile, the applications and developments of analytical methods of polar small molecules, such as reversed-phase liquid chromatography, hydrophilic interaction chromatography, etc., are also reviewed.

  19. [Optimized sample preparation for metabolome studies on Streptomyces coelicolor].

    PubMed

    Li, Yihong; Li, Shanshan; Ai, Guomin; Wang, Weishan; Zhang, Buchang; Yang, Keqian

    2014-04-01

    Streptomycetes produce many antibiotics and are important model microorgansims for scientific research and antibiotic production. Metabolomics is an emerging technological platform to analyze low molecular weight metabolites in a given organism qualitatively and quantitatively. Compared to other Omics platform, metabolomics has greater advantage in monitoring metabolic flux distribution and thus identifying key metabolites related to target metabolic pathway. The present work aims at establishing a rapid, accurate sample preparation protocol for metabolomics analysis in streptomycetes. In the present work, several sample preparation steps, including cell quenching time, cell separation method, conditions for metabolite extraction and metabolite derivatization were optimized. Then, the metabolic profiles of Streptomyces coelicolor during different growth stages were analyzed by GC-MS. The optimal sample preparation conditions were as follows: time of low-temperature quenching 4 min, cell separation by fast filtration, time of freeze-thaw 45 s/3 min and the conditions of metabolite derivatization at 40 degrees C for 90 min. By using this optimized protocol, 103 metabolites were finally identified from a sample of S. coelicolor, which distribute in central metabolic pathways (glycolysis, pentose phosphate pathway and citrate cycle), amino acid, fatty acid, nucleotide metabolic pathways, etc. By comparing the temporal profiles of these metabolites, the amino acid and fatty acid metabolic pathways were found to stay at a high level during stationary phase, therefore, these pathways may play an important role during the transition between the primary and secondary metabolism. An optimized protocol of sample preparation was established and applied for metabolomics analysis of S. coelicolor, 103 metabolites were identified. The temporal profiles of metabolites reveal amino acid and fatty acid metabolic pathways may play an important role in the transition from primary to

  20. Application of hydrocyanic acid vapor generation via focused microwave radiation to the preparation of industrial effluent samples prior to free and total cyanide determinations by spectrophotometric flow injection analysis.

    PubMed

    Quaresma, Maria Cristina Baptista; de Carvalho, Maria de Fátima Batista; Meirelles, Francis Assis; Santiago, Vânia Maria Junqueira; Santelli, Ricardo Erthal

    2007-02-01

    A sample preparation procedure for the quantitative determination of free and total cyanides in industrial effluents has been developed that involves hydrocyanic acid vapor generation via focused microwave radiation. Hydrocyanic acid vapor was generated from free cyanides using only 5 min of irradiation time (90 W power) and a purge time of 5 min. The HCN generated was absorbed into an accepting NaOH solution using very simple glassware apparatus that was appropriate for the microwave oven cavity. After that, the cyanide concentration was determined within 90 s using a well-known spectrophotometric flow injection analysis system. Total cyanide analysis required 15 min irradiation time (90 W power), as well as chemical conditions such as the presence of EDTA-acetate buffer solution or ascorbic acid, depending on the effluent to be analyzed (petroleum refinery or electroplating effluents, respectively). The detection limit was 0.018 mg CN l(-1) (quantification limit of 0.05 mg CN l(-1)), and the measured RSD was better than 8% for ten independent analyses of effluent samples (1.4 mg l(-1) cyanide). The accuracy of the procedure was assessed via analyte spiking (with free and complex cyanides) and by performing an independent sample analysis based on the standard methodology recommended by the APHA for comparison. The sample preparation procedure takes only 10 min for free and 20 min for total cyanide, making this procedure much faster than traditional methodologies (conventional heating and distillation), which are time-consuming (they require at least 1 h). Samples from oil (sour and stripping tower bottom waters) and electroplating effluents were analyzed successfully.

  1. HASE - The Helsinki adaptive sample preparation line

    NASA Astrophysics Data System (ADS)

    Palonen, V.; Pesonen, A.; Herranen, T.; Tikkanen, P.; Oinonen, M.

    2013-01-01

    We have designed and built an adaptive sample preparation line with separate modules for combustion, molecular sieve handling, CO2 gas cleaning, CO2 storage, and graphitization. The line is also connected to an elemental analyzer. Operation of the vacuum equipment, a flow controller, pressure sensors, ovens, and graphitization reactors are automated with a reliable NI-cRIO real-time system. Stepped combustion can be performed in two ovens at temperatures up to 900 °C. Depending on the application, CuO or O2-flow combustion can be used. A flow controller is used to adjust the O2 flow and pressure during combustion. For environmental samples, a module for molecular sieve regeneration and sample desorption is attached to the line replacing the combustion module. In the storage module, CO2 samples can be stored behind a gas-tight diaphragm valve and either stored for later graphitization or taken for measurements with separate equipment (AMS gas ion source or a separate mass spectrometer). The graphitization module consists of four automated reactors, capable of graphitizing samples with masses from 3 mg down to 50 μg.

  2. Amino acid analyses of Apollo 14 samples.

    NASA Technical Reports Server (NTRS)

    Gehrke, C. W.; Zumwalt, R. W.; Kuo, K.; Aue, W. A.; Stalling, D. L.; Kvenvolden, K. A.; Ponnamperuma, C.

    1972-01-01

    Detection limits were between 300 pg and 1 ng for different amino acids, in an analysis by gas-liquid chromatography of water extracts from Apollo 14 lunar fines in which amino acids were converted to their N-trifluoro-acetyl-n-butyl esters. Initial analyses of water and HCl extracts of sample 14240 and 14298 samples showed no amino acids above background levels.

  3. Chapter A1. Preparations for Water Sampling

    USGS Publications Warehouse

    Wilde, Franceska D.; Radtke, Dean B.; Gibs, Jacob; Iwatsubo, Rick T.

    1998-01-01

    The National Field Manual for the Collection of Water-Quality Data (National Field Manual) provides guidelines and standard procedures for U.S. Geological Survey (USGS) personnel who collect data used to assess the quality of the Nation's surface-water and ground-water resources. This chapter addresses field-trip preparations, including selection of sample-collection sites for studies of surface-water quality, site reconnaissance and well selection for studies of groundwater quality, and the establishment of electronic files and field files and folders for a sampling site. Each chapter of the National Field Manual is published separately and revised periodically. Newly published and revised chapters are posted on the World Wide Web on the USGS page 'National Field Manual for the Collection of Water-Quality Data.' The URL for this page is http://pubs.water.usgs.gov/twri9A/ (accessed Jan. 31, 2005).

  4. Microextraction sample preparation techniques in biomedical analysis.

    PubMed

    Szultka, Malgorzata; Pomastowski, Pawel; Railean-Plugaru, Viorica; Buszewski, Boguslaw

    2014-11-01

    Biologically active compounds are found in biological samples at relatively low concentration levels. The sample preparation of target compounds from biological, pharmaceutical, environmental, and food matrices is one of the most time-consuming steps in the analytical procedure. The microextraction techniques are dominant. Metabolomic studies also require application of proper analytical technique for the determination of endogenic metabolites present in biological matrix on trace concentration levels. Due to the reproducibility of data, precision, relatively low cost of the appropriate analysis, simplicity of the determination, and the possibility of direct combination of those techniques with other methods (combination types on-line and off-line), they have become the most widespread in routine determinations. Additionally, sample pretreatment procedures have to be more selective, cheap, quick, and environmentally friendly. This review summarizes the current achievements and applications of microextraction techniques. The main aim is to deal with the utilization of different types of sorbents for microextraction and emphasize the use of new synthesized sorbents as well as to bring together studies concerning the systematic approach to method development. This review is dedicated to the description of microextraction techniques and their application in biomedical analysis.

  5. 7 CFR 27.21 - Preparation of samples of cotton.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Preparation of samples of cotton. 27.21 Section 27.21... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Inspection and Samples § 27.21 Preparation of samples of cotton. The samples from each bale shall be prepared as specified in this...

  6. 7 CFR 27.21 - Preparation of samples of cotton.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Preparation of samples of cotton. 27.21 Section 27.21... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Inspection and Samples § 27.21 Preparation of samples of cotton. The samples from each bale shall be prepared as specified in this...

  7. 7 CFR 27.21 - Preparation of samples of cotton.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Preparation of samples of cotton. 27.21 Section 27.21... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Inspection and Samples § 27.21 Preparation of samples of cotton. The samples from each bale shall be prepared as specified in this...

  8. Sampling almonds for aflatoxin, part I: estimation of uncertainty associated with sampling, sample preparation, and analysis.

    PubMed

    Whitaker, Thomas B; Slate, Andrew B; Jacobs, Merle; Hurley, J Michael; Adams, Julie G; Giesbrecht, Francis G

    2006-01-01

    Domestic and international regulatory limits have been established for aflatoxin in almonds and other tree nuts. It is difficult to obtain an accurate and precise estimate of the true aflatoxin concentration in a bulk lot because of the uncertainty associated with the sampling, sample preparation, and analytical steps of the aflatoxin test procedure. To evaluate the performance of aflatoxin sampling plans, the uncertainty associated with sampling lots of shelled almonds for aflatoxin was investigated. Twenty lots of shelled almonds were sampled for aflatoxin contamination. The total variance associated with measuring B1 and total aflatoxins in bulk almond lots was estimated and partitioned into sampling, sample preparation, and analytical variance components. All variances were found to increase with an increase in aflatoxin concentration (both B1 and total). By using regression analysis, mathematical expressions were developed to predict the relationship between each variance component (total, sampling, sample preparation, and analysis variances) and aflatoxin concentration. Variance estimates were the same for B1 and total aflatoxins. The mathematical relationships can be used to estimate each variance for a given sample size, subsample size, and number of analyses other than that measured in the study. When a lot with total aflatoxins at 15 ng/g was tested by using a 10 kg sample, a vertical cutter mixer type of mill, a 100 g subsample, and high-performance liquid chromatography analysis, the sampling, sample preparation, analytical, and total variances (coefficient of variation, CV) were 394.7 (CV, 132.4%), 14.7 (CV, 25.5%), 0.8 (CV, 6.1%), and 410.2 (CV, 135.0%), respectively. The percentages of the total variance associated with sampling, sample preparation, and analytical steps were 96.2, 3.6, and 0.2, respectively.

  9. Sampling hazelnuts for aflatoxin: uncertainty associated with sampling, sample preparation, and analysis.

    PubMed

    Ozay, Guner; Seyhan, Ferda; Yilmaz, Aysun; Whitaker, Thomas B; Slate, Andrew B; Giesbrecht, Francis

    2006-01-01

    The variability associated with the aflatoxin test procedure used to estimate aflatoxin levels in bulk shipments of hazelnuts was investigated. Sixteen 10 kg samples of shelled hazelnuts were taken from each of 20 lots that were suspected of aflatoxin contamination. The total variance associated with testing shelled hazelnuts was estimated and partitioned into sampling, sample preparation, and analytical variance components. Each variance component increased as aflatoxin concentration (either B1 or total) increased. With the use of regression analysis, mathematical expressions were developed to model the relationship between aflatoxin concentration and the total, sampling, sample preparation, and analytical variances. The expressions for these relationships were used to estimate the variance for any sample size, subsample size, and number of analyses for a specific aflatoxin concentration. The sampling, sample preparation, and analytical variances associated with estimating aflatoxin in a hazelnut lot at a total aflatoxin level of 10 ng/g and using a 10 kg sample, a 50 g subsample, dry comminution with a Robot Coupe mill, and a high-performance liquid chromatographic analytical method are 174.40, 0.74, and 0.27, respectively. The sampling, sample preparation, and analytical steps of the aflatoxin test procedure accounted for 99.4, 0.4, and 0.2% of the total variability, respectively.

  10. Preparation of stir cake sorptive extraction based on poly(4-vinylbenzoic acid-divinylbenzene) monolith and its application in sensitive determination of β-agonists in milk and swine urine samples.

    PubMed

    Huang, Xiaojia; Chen, Linli; Yuan, Dongxing

    2013-11-15

    In this study, a new stir cake sorptive extraction (SCSE) based on poly(4-vinylbenzoic acid-divinylbenzene) (VBADB) monolith was prepared. The effect of preparation conditions of monolith on extraction efficiencies was investigated in detail. Several characteristic techniques, such as elemental analysis, infrared spectroscopy, mercury intrusion porosimetry and scanning electron microscopy were used to characterize the monolithic material. The combination of SCSE-VBADB with high-performance liquid chromatography tandem mass spectrometry (HPLC/MS/MS) detection was developed for sensitive determination of ultra-trace β-agonists in milk and swine urine samples. In order to obtain the optimal extraction conditions of SCSE-VBADB for β-agonists, several extractive parameters, including pH values and ionic strength in sample matrix, extraction and desorption time were optimized. Under the optimum conditions, the limits of detection (S/N=3) for the target analytes were 0.007-0.030 μg/L in milk and 0.002-0.011 μg/L in swine urine, respectively. Excellent method reproducibility was achieved in terms of intraday and interday precisions, indicated by the RSDs of both <10.0%, respectively. Finally, the proposed method was successfully used to detect β-agonists in different milk and swine urines samples. Acceptable recoveries ranged from 50.3% to 113% and 50.1% to 92.2% for milk and swine urine samples, respectively; and the RSDs for reproducibility were less than 8.0% for target analytes in all real samples.

  11. Study on dicarboxylic acids in aerosol samples with capillary electrophoresis.

    PubMed

    Adler, Heidi; Sirén, Heli

    2014-01-01

    The research was performed to study the simultaneous detection of a homologous series of α , ω -dicarboxylic acids (C2-C10), oxalic, malonic, succinic, glutaric, adipic, pimelic, suberic, azelaic, and sebacic acids, with capillary electrophoresis using indirect UV detection. Good separation efficiency in 2,6-pyridinedicarboxylic acid as background electrolyte modified with myristyl trimethyl ammonium bromide was obtained. The dicarboxylic acids were ionised and separated within five minutes. For the study, authentic samples were collected onto dry cellulose membrane filters of a cascade impactor (12 stages) from outdoor spring aerosols in an urban area. Hot water and ultrasonication extraction methods were used to isolate the acids from membrane filters. Due to the low concentrations of acids in the aerosols, the extracts were concentrated with solid-phase extraction (SPE) before determination. The enrichment of the carboxylic acids was between 86 and 134% with sample pretreatment followed by 100-time increase by preparation of the sample to 50  μ L. Inaccuracy was optimised for all the sample processing steps. The aerosols contained dicarboxylic acids C2-C10. Then, mostly they contained C2, C5, and C10. Only one sample contained succinic acid. In the study, the concentrations of the acids in aerosols were lower than 10 ng/m(3).

  12. Preparation of graphene thin films for radioactive samples.

    PubMed

    Roteta, Miguel; Fernández-Martínez, Rodolfo; Mejuto, Marcos; Rucandio, Isabel

    2016-03-01

    A new method for the preparation of conductive thin films is presented. The metallization of VYNS films guarantees the electrical conductivity but it results in the breaking of a high proportion of them. Graphene, a two-dimensional nanostructure of monolayer or few layers graphite has attracted a great deal of attention because of its excellent properties such as a good chemical stability, mechanical resistance and extraordinary electronic transport properties. In this work, the possibilities of graphene have been explored as a way to produce electrical conductive thin films without an extra metallization process. The procedure starts with preparing homogenous suspensions of reduced graphene oxide (rGO) in conventional VYNS solutions. Ultra-sonication is used to ensure a good dispersibility of rGO. Graphene oxide (GO) is prepared via oxidation of graphite and subsequent exfoliation by sonication. Different chemically rGO were obtained by reaction with hydrazine sulfate, sodium borohydride, ascorbic acid and hydroiodic acid as reducing agents. The preparation of the thin graphene films is done in a similar way as the conventional VYNS foil preparation procedure. Drops of the solution are deposited onto water. The graphene films have been used to prepare sources containing some electron capture radionuclides ((109)Cd, (55)Fe, (139)Ce) with an activity in the order of 3kBq. The samples have been measured to test the attainable low energy electron efficiency and the energy resolution of Auger and conversion electrons by 4π (electron capture)-γ coincidence measurements. The 4π (electron capture)-γ coincidence setup includes a pressurized proportional counter and a NaI(Tl) detector. Tests with different pressures up to 1000kPa were carried out. All these tests show similar values in both parameters (efficiency and resolution) as those obtained by using the conventional metallized films without the drawback of the high percentage of broken films.

  13. Preparation of urine samples for proteomic analysis.

    PubMed

    Pieper, Rembert

    2008-01-01

    Reproducible procedures for the preparation of protein samples isolated from human urine are essential for meaningful proteomic analyses. Key applications are the discovery of novel proteins or their modifications in the human urine as well as protein biomarker discovery for diseases and drug treatments. The methodology presented here features experimental steps aimed at limiting protein losses because of organic solvent precipitation, effective separation of proteins from other compounds in the human urine and molecular weight-based enrichment of proteins in two distinct fractions. Urinary proteins are separated from cellular debris in the urine via centrifugation, concentrated with 5-kDa-cutoff membrane concentration devices and separated via size exclusion chromatography into fractions with a higher and a lower molecular weight than 30 kDa, respectively. A successive optional affinity removal step for highly abundant plasma proteins is described. Finally, buffer exchange steps useful for specific downstream proteomic analysis experiments of urinary proteins are presented, such as 2-dimensional gel electrophoresis, differential protein or peptide labeling and digestion with trypsin for LC-MS/MS analysis.

  14. Microfluidic DNA sample preparation method and device

    DOEpatents

    Krulevitch, Peter A.; Miles, Robin R.; Wang, Xiao-Bo; Mariella, Raymond P.; Gascoyne, Peter R. C.; Balch, Joseph W.

    2002-01-01

    Manipulation of DNA molecules in solution has become an essential aspect of genetic analyses used for biomedical assays, the identification of hazardous bacterial agents, and in decoding the human genome. Currently, most of the steps involved in preparing a DNA sample for analysis are performed manually and are time, labor, and equipment intensive. These steps include extraction of the DNA from spores or cells, separation of the DNA from other particles and molecules in the solution (e.g. dust, smoke, cell/spore debris, and proteins), and separation of the DNA itself into strands of specific lengths. Dielectrophoresis (DEP), a phenomenon whereby polarizable particles move in response to a gradient in electric field, can be used to manipulate and separate DNA in an automated fashion, considerably reducing the time and expense involved in DNA analyses, as well as allowing for the miniaturization of DNA analysis instruments. These applications include direct transport of DNA, trapping of DNA to allow for its separation from other particles or molecules in the solution, and the separation of DNA into strands of varying lengths.

  15. Waste minimization in analytical chemistry through innovative sample preparation techniques.

    SciTech Connect

    Smith, L. L.

    1998-05-28

    Because toxic solvents and other hazardous materials are commonly used in analytical methods, characterization procedures result in significant and costly amount of waste. We are developing alternative analytical methods in the radiological and organic areas to reduce the volume or form of the hazardous waste produced during sample analysis. For the radiological area, we have examined high-pressure, closed-vessel microwave digestion as a way to minimize waste from sample preparation operations. Heated solutions of strong mineral acids can be avoided for sample digestion by using the microwave approach. Because reactivity increases with pressure, we examined the use of less hazardous solvents to leach selected contaminants from soil for subsequent analysis. We demonstrated the feasibility of this approach by extracting plutonium from a NET reference material using citric and tartaric acids with microwave digestion. Analytical results were comparable to traditional digestion methods, while hazardous waste was reduced by a factor often. We also evaluated the suitability of other natural acids, determined the extraction performance on a wider variety of soil types, and examined the extraction efficiency of other contaminants. For the organic area, we examined ways to minimize the wastes associated with the determination of polychlorinated biphenyls (PCBs) in environmental samples. Conventional methods for analyzing semivolatile organic compounds are labor intensive and require copious amounts of hazardous solvents. For soil and sediment samples, we have a method to analyze PCBs that is based on microscale extraction using benign solvents (e.g., water or hexane). The extraction is performed at elevated temperatures in stainless steel cells containing the sample and solvent. Gas chromatography-mass spectrometry (GC/MS) was used to quantitate the analytes in the isolated extract. More recently, we developed a method utilizing solid-phase microextraction (SPME) for natural

  16. Spectral Reproducibility and Quantification of Peptides in MALDI of Samples Prepared by Micro-Spotting

    NASA Astrophysics Data System (ADS)

    Bae, Yong Jin; Park, Kyung Man; Ahn, Sung Hee; Moon, Jeong Hee; Kim, Myung Soo

    2014-08-01

    Previously, we reported that MALDI spectra of peptides became reproducible when temperature was kept constant. Linear calibration curves derived from such spectral data could be used for quantification. Homogeneity of samples was one of the requirements. Among the three popular matrices used in peptide MALDI [i.e., α-cyano-4-hydroxycinnamic acid (CHCA), 2,5-dihydroxybenzoic acid (DHB), and sinapinic acid (SA)], homogeneous samples could be prepared by conventional means only for CHCA. In this work, we showed that sample preparation by micro-spotting improved the homogeneity for all three cases.

  17. Research for amino acids in lunar samples.

    NASA Technical Reports Server (NTRS)

    Gehrke, C. W.; Zumwalt, R. W.; Kuo, K.; Rash, J. J.; Aue , W. A.; Stalling, D. L.; Kvenvolden, K. A.; Ponnamperuma, C.

    1972-01-01

    The study was primarily directed toward the examination of Apollo 14 lunar fines for indigenous amino acids or materials which could be converted to amino acids on hydrolysis with 6 N hydrochloric acid. Initial experiments were conducted to confirm the integrity of the derivatization reactions and reagents, and to optimize the gas-liquid chromatographic (GLC) instrumental and chromatographic system for the separation and flame ionization detection of the amino acid derivatives. In studies on the recovery of amino acids added to lunar fines, low recoveries were obtained when 10 ng of each amino acid were added to 50 mg of virgin fines, but the subsequent addition of 50 ng of each to the previously extracted sample resulted in much higher recoveries.

  18. Preparation of Cytology Samples: Tricks of the Trade.

    PubMed

    Moore, A Russell

    2017-01-01

    General principles and techniques for collection, preparation, and staining of cytologic samples in the general practice setting are reviewed. Tips for collection of digital images are also discussed.

  19. Components for automated microfluidics sample preparation and analysis

    NASA Astrophysics Data System (ADS)

    Archer, M.; Erickson, J. S.; Hilliard, L. R.; Howell, P. B., Jr.; Stenger, D. A.; Ligler, F. S.; Lin, B.

    2008-02-01

    The increasing demand for portable devices to detect and identify pathogens represents an interdisciplinary effort between engineering, materials science, and molecular biology. Automation of both sample preparation and analysis is critical for performing multiplexed analyses on real world samples. This paper selects two possible components for such automated portable analyzers: modified silicon structures for use in the isolation of nucleic acids and a sheath flow system suitable for automated microflow cytometry. Any detection platform that relies on the genetic content (RNA and DNA) present in complex matrices requires careful extraction and isolation of the nucleic acids in order to ensure their integrity throughout the process. This sample pre-treatment step is commonly performed using commercially available solid phases along with various molecular biology techniques that require multiple manual steps and dedicated laboratory space. Regardless of the detection scheme, a major challenge in the integration of total analysis systems is the development of platforms compatible with current isolation techniques that will ensure the same quality of nucleic acids. Silicon is an ideal candidate for solid phase separations since it can be tailored structurally and chemically to mimic the conditions used in the laboratory. For analytical purposes, we have developed passive structures that can be used to fully ensheath one flow stream with another. As opposed to traditional flow focusing methods, our sheath flow profile is truly two dimensional, making it an ideal candidate for integration into a microfluidic flow cytometer. Such a microflow cytometer could be used to measure targets captured on either antibody- or DNA-coated beads.

  20. Rapid Automated Sample Preparation for Biological Assays

    SciTech Connect

    Shusteff, M

    2011-03-04

    Our technology utilizes acoustic, thermal, and electric fields to separate out contaminants such as debris or pollen from environmental samples, lyse open cells, and extract the DNA from the lysate. The objective of the project is to optimize the system described for a forensic sample, and demonstrate its performance for integration with downstream assay platforms (e.g. MIT-LL's ANDE). We intend to increase the quantity of DNA recovered from the sample beyond the current {approx}80% achieved using solid phase extraction methods. Task 1: Develop and test an acoustic filter for cell extraction. Task 2: Develop and test lysis chip. Task 3: Develop and test DNA extraction chip. All chips have been fabricated based on the designs laid out in last month's report.

  1. 40 CFR 761.392 - Preparing validation study samples.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 31 2014-07-01 2014-07-01 false Preparing validation study samples..., AND USE PROHIBITIONS Comparison Study for Validating a New Performance-Based Decontamination Solvent Under § 761.79(d)(4) § 761.392 Preparing validation study samples. (a)(1) To validate a procedure...

  2. 40 CFR 761.392 - Preparing validation study samples.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false Preparing validation study samples..., AND USE PROHIBITIONS Comparison Study for Validating a New Performance-Based Decontamination Solvent Under § 761.79(d)(4) § 761.392 Preparing validation study samples. (a)(1) To validate a procedure...

  3. 40 CFR 761.392 - Preparing validation study samples.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false Preparing validation study samples..., AND USE PROHIBITIONS Comparison Study for Validating a New Performance-Based Decontamination Solvent Under § 761.79(d)(4) § 761.392 Preparing validation study samples. (a)(1) To validate a procedure...

  4. Ultra-Fast Sample Preparation for High-Throughput Proteomics

    SciTech Connect

    Lopez-Ferrer, Daniel; Hixson, Kim K.; Belov, Mikhail E.; Smith, Richard D.

    2011-06-21

    Sample preparation oftentimes can be the Achilles Heel of any analytical process and in the field of proteomics, preparing samples for mass spectrometric analysis is no exception. Current goals, concerning proteomic sample preparation on a large scale, include efforts toward improving reproducibility, reducing the time of processing and ultimately the automation of the entire workflow. This chapter reviews an array of recent approaches applied to bottom-up proteomics sample preparation to reduce the processing time down from hours to minutes. The current state-of-the-art in the field uses different energy inputs like microwave, ultrasound or pressure to perform the four basic steps in sample preparation: protein extraction, denaturation, reduction and alkylation, and digestion. No single energy input for enhancement of proteome sample preparation has become the universal gold standard. Instead, a combination of different energy inputs tend to produce the best results. This chapter further describes the future trends in the field such as the hyphenation of sample preparation with downstream detection and analysis systems. Finally, a detailed protocol describing the combined use of both pressure cycling technology and ultrasonic energy inputs to hasten proteomic sample preparation is presented.

  5. DICARBOXYLIC ACID CONCENTRATION TRENDS AND SAMPLING ARTIFACTS

    EPA Science Inventory

    Dicarboxylic acids associated with airborne particulate matter were measured during a summer period in Philadelphia that included multiple air pollution episodes. Samples were collected for two ten hour periods each day using a high volume sampler with two quartz fiber filters in...

  6. Potential artifacts associated with historical preparation of joint compound samples and reported airborne asbestos concentrations.

    PubMed

    Brorby, G P; Sheehan, P J; Berman, D W; Bogen, K T; Holm, S E

    2011-05-01

    Airborne samples collected in the 1970s for drywall workers using asbestos-containing joint compounds were likely prepared and analyzed according to National Institute of Occupational Safety and Health Method P&CAM 239, the historical precursor to current Method 7400. Experimentation with a re-created, chrysotile-containing, carbonate-based joint compound suggested that analysis following sample preparation by the historical vs. current method produces different fiber counts, likely because of an interaction between the different clearing and mounting chemicals used and the carbonate-based joint compound matrix. Differences were also observed during analysis using Method 7402, depending on whether acetic acid/dimethylformamide or acetone was used during preparation to collapse the filter. Specifically, air samples of sanded chrysotile-containing joint compound prepared by the historical method yielded fiber counts significantly greater (average of 1.7-fold, 95% confidence interval: 1.5- to 2.0-fold) than those obtained by the current method. In addition, air samples prepared by Method 7402 using acetic acid/dimethylformamide yielded fiber counts that were greater (2.8-fold, 95% confidence interval: 2.5- to 3.2-fold) than those prepared by this method using acetone. These results indicated (1) there is an interaction between Method P&CAM 239 preparation chemicals and the carbonate-based joint compound matrix that reveals fibers that were previously bound in the matrix, and (2) the same appeared to be true for Method 7402 preparation chemicals acetic acid/dimethylformamide. This difference in fiber counts is the opposite of what has been reported historically for samples of relatively pure chrysotile dusts prepared using the same chemicals. This preparation artifact should be considered when interpreting historical air samples for drywall workers prepared by Method P&CAM 239.

  7. Snow White Trench Prepared for Sample Collection

    NASA Technical Reports Server (NTRS)

    2008-01-01

    The informally named 'Snow White' trench is the source for the next sample to be acquired by NASA's Phoenix Mars Lander for analysis by the wet chemistry lab.

    The Surface Stereo Imager on Phoenix took this shadow-enhanced image of the trench, on the eastern end of Phoenix's work area, on Sol 103, or the 103rd day of the mission, Sept. 8, 2008. The trench is about 23 centimeters (9 inches) wide.

    The wet chemistry lab is part of Phoenix's Microscopy, Electrochemistry and Conductivity suite of instruments.

    The Phoenix Mission is led by the University of Arizona, Tucson, on behalf of NASA. Project management of the mission is by NASA's Jet Propulsion Laboratory, Pasadena, Calif. Spacecraft development is by Lockheed Martin Space Systems, Denver.

  8. Preparation of ultra small samples for optical and microprobe analysis

    NASA Technical Reports Server (NTRS)

    Inman, C. S.

    1973-01-01

    This paper describes a simple but satisfactory new method for the preparation of tiny, varied and specialized specimens for electron or ion-microprobe analysis developed over the past five years. Microtektites, individual chondrules, single grains, blebs from lunar samples and meteoritic minerals have been prepared by this technique. A description of the preparation of these usually difficult samples from the initial mounting through the various polishing steps to their final polish is presented in detail. The procedures used to prevent any contamination of these specimens by the polishing agents and to prevent cross contamination to the other samples used for geochronology studies are presented.

  9. Free amino acids in botanicals and botanical preparations.

    PubMed

    Carratù, B; Boniglia, C; Giammarioli, S; Mosca, M; Sanzini, E

    2008-06-01

    Numerous studies were carried out about aminoacidic composition of vegetable proteins, but information about the free amino acid pool and the role of these substances is very incomplete. The aim of this paper was to contribute to the scarce knowledge concerning the composition of free amino acids in botanicals and botanical preparations widely used as food, in dietary supplements, and in pharmaceutical products. This work studied the composition of free amino acids, identified the major components of 19 species of plants, and evaluated the influence of different types of extraction on the amino acid profile. Amino acids were determined using an automatic precolumn derivatization with fluorenylmethyl-chloroformate and reversed-phase liquid chromatography with fluorescence and ultraviolet detection. The amounts of total free amino acids varied widely between plants, from approximately 12 g in 100 g of Echinacea pallida extract to less than 60 mg in the same amount of Coleus forskohlii, Garcinia cambogia, and Glycine max. In 13 plants arginine, asparagine, glutamine, proline, and gamma-aminobutyric acid were the free amino acids found in preponderant quantities. The levels of free amino acids above the quantification limit in 36 assayed samples of botanicals, extracts, and supplements are shown.

  10. Curatorial Works for the Hayabusa-Returned Sample and Preparation for Hayabusa2 Sample Curation

    NASA Astrophysics Data System (ADS)

    Yada, T.; Abe, M.; Okada, T.; Yurimoto, H.; Uesugi, M.; Karouji, Y.; Nakato, A.; Hashiguchi, M.; Nishimura, M.; Kumagai, K.; Matsui, S.; Yoshitake, M.; Sakamoto, K.; Nakano, Y.; Kawasaki, N.; Fujimoto, M.

    2016-08-01

    We continue describing Hayabusa-returned samples after its return in 2010. The number of described particles reaches around 650 and >540 of them are identified as Itokawa origin. We also start preparation for Hayabusa2 sample curation.

  11. Novel Sample-handling Approach for XRD Analysis with Minimal Sample Preparation

    NASA Technical Reports Server (NTRS)

    Sarrazin, P.; Chipera, S.; Bish, D.; Blake, D.; Feldman, S.; Vaniman, D.; Bryson, C.

    2004-01-01

    Sample preparation and sample handling are among the most critical operations associated with X-ray diffraction (XRD) analysis. These operations require attention in a laboratory environment, but they become a major constraint in the deployment of XRD instruments for robotic planetary exploration. We are developing a novel sample handling system that dramatically relaxes the constraints on sample preparation by allowing characterization of coarse-grained material that would normally be impossible to analyze with conventional powder-XRD techniques.

  12. Preparation and bactericide activity of gallic acid stabilized gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Moreno-Álvarez, S. A.; Martínez-Castañón, G. A.; Niño-Martínez, N.; Reyes-Macías, J. F.; Patiño-Marín, N.; Loyola-Rodríguez, J. P.; Ruiz, Facundo

    2010-10-01

    In this work, gold nanoparticles with three different sizes (13.7, 39.4, and 76.7 nm) were prepared using a simple aqueous method with gallic acid as the reducing and stabilizing agent, the different sizes were obtained varying some experimental parameters as the pH of the reaction and the amount of the gallic acid. The prepared nanoparticles were characterized using X-ray diffraction, transmission electron microscopy, dynamic light scattering, and UV-Vis spectroscopy. Samples were identified as elemental gold and present spherical morphology, a narrow size distribution and good stabilization according to TEM and DLS results. The antibacterial activity of this gallic acid stabilized gold nanoparticles against S. mutans (the etiologic agent of dental caries) was assessed using a microdilution method obtaining a minimum inhibitory concentration of 12.31, 12.31, and 49.25 μg/mL for 13.7, 39.4, and 76.7 nm gold nanoparticles, respectively. The antibacterial assay showed that gold nanoparticles prepared in this work present a bactericide activity by a synergistic action with gallic acid. The MIC found for this nanoparticles are much lower than those reported for mixtures of gold nanoparticles and antibiotics.

  13. Analytical Chemistry Laboratory (ACL) procedure compendium. Volume 2, Sample preparation methods

    SciTech Connect

    Not Available

    1993-08-01

    This volume contains the interim change notice for sample preparation methods. Covered are: acid digestion for metals analysis, fusion of Hanford tank waste solids, water leach of sludges/soils/other solids, extraction procedure toxicity (simulate leach in landfill), sample preparation for gamma spectroscopy, acid digestion for radiochemical analysis, leach preparation of solids for free cyanide analysis, aqueous leach of solids for anion analysis, microwave digestion of glasses and slurries for ICP/MS, toxicity characteristic leaching extraction for inorganics, leach/dissolution of activated metal for radiochemical analysis, extraction of single-shell tank (SST) samples for semi-VOC analysis, preparation and cleanup of hydrocarbon- containing samples for VOC and semi-VOC analysis, receiving of waste tank samples in onsite transfer cask, receipt and inspection of SST samples, receipt and extrusion of core samples at 325A shielded facility, cleaning and shipping of waste tank samplers, homogenization of solutions/slurries/sludges, and test sample preparation for bioassay quality control program.

  14. Preparation of privatization samples for envelopes `A` and `C`

    SciTech Connect

    Winters, W.I., Westinghouse Hanford

    1996-07-17

    As part of the TWRS Privatization process, the DOE has committed to provide each of the two contractors who submitted successful bids with ten 125 mL samples of Hanford tank waste meeting chemical and radionuclide criteria specified as Waste Envelope A, B, and C. This test plan describes how the samples will be prepared before shipment.

  15. Preparation of bone samples in the Gliwice Radiocarbon Laboratory for AMS radiocarbon dating.

    PubMed

    Piotrowska, N; Goslar, T

    2002-12-01

    In the Gliwice Radiocarbon Laboratory, a system for preparation of samples for AMS dating has been built. At first it was used to produce graphite targets from plant macrofossils and sediments. In this study we extended its capabilities with the preparation of bones. We dealt with 3 methods; the first was the classical Longin method of collagen extraction, the second one included additional treatment of powdered bone in alkali solution, while in the third one carboxyl carbon was separated from amino acids obtained after hydrolysis of protein. The suitability of the methods was tested on 2 bone samples. Most of our samples gave ages > 40 kyr BP, suggesting good performance of the adapted methods, except for one sample prepared with simple Longin method. For routine preparation of bones we chose the Longin method with additional alkali treatment.

  16. New materials for sample preparation techniques in bioanalysis.

    PubMed

    Nazario, Carlos Eduardo Domingues; Fumes, Bruno Henrique; da Silva, Meire Ribeiro; Lanças, Fernando Mauro

    2017-02-01

    The analysis of biological samples is a complex and difficult task owing to two basic and complementary issues: the high complexity of most biological matrices and the need to determine minute quantities of active substances and contaminants in such complex sample. To succeed in this endeavor samples are usually subject to three steps of a comprehensive analytical methodological approach: sample preparation, analytes isolation (usually utilizing a chromatographic technique) and qualitative/quantitative analysis (usually with the aid of mass spectrometric tools). Owing to the complex nature of bio-samples, and the very low concentration of the target analytes to be determined, selective sample preparation techniques is mandatory in order to overcome the difficulties imposed by these two constraints. During the last decade new chemical synthesis approaches has been developed and optimized, such as sol-gel and molecularly imprinting technologies, allowing the preparation of novel materials for sample preparation including graphene and derivatives, magnetic materials, ionic liquids, molecularly imprinted polymers, and much more. In this contribution we will review these novel techniques and materials, as well as their application to the bioanalysis niche.

  17. Automation of preparation of nonmetallic samples for analysis by atomic absorption and inductively coupled plasma spectrometry

    NASA Technical Reports Server (NTRS)

    Wittmann, A.; Willay, G.

    1986-01-01

    For a rapid preparation of solutions intended for analysis by inductively coupled plasma emission spectrometry or atomic absorption spectrometry, an automatic device called Plasmasol was developed. This apparatus used the property of nonwettability of glassy C to fuse the sample in an appropriate flux. The sample-flux mixture is placed in a composite crucible, then heated at high temperature, swirled until full dissolution is achieved, and then poured into a water-filled beaker. After acid addition, dissolution of the melt, and filling to the mark, the solution is ready for analysis. The analytical results obtained, either for oxide samples or for prereduced iron ores show that the solutions prepared with this device are undistinguished from those obtained by manual dissolutions done by acid digestion or by high temperature fusion. Preparation reproducibility and analytical tests illustrate the performance of Plasmasol.

  18. Advanced Curation Preparation for Mars Sample Return and Cold Curation

    NASA Technical Reports Server (NTRS)

    Fries, M. D.; Harrington, A. D.; McCubbin, F. M.; Mitchell, J.; Regberg, A. B.; Snead, C.

    2017-01-01

    NASA Curation is tasked with the care and distribution of NASA's sample collections, such as the Apollo lunar samples and cometary material collected by the Stardust spacecraft. Curation is also mandated to perform Advanced Curation research and development, which includes improving the curation of existing collections as well as preparing for future sample return missions. Advanced Curation has identified a suite of technologies and techniques that will require attention ahead of Mars sample return (MSR) and missions with cold curation (CCur) requirements, perhaps including comet sample return missions.

  19. Magnetic separation techniques in sample preparation for biological analysis: a review.

    PubMed

    He, Jincan; Huang, Meiying; Wang, Dongmei; Zhang, Zhuomin; Li, Gongke

    2014-12-01

    Sample preparation is a fundamental and essential step in almost all the analytical procedures, especially for the analysis of complex samples like biological and environmental samples. In past decades, with advantages of superparamagnetic property, good biocompatibility and high binding capacity, functionalized magnetic materials have been widely applied in various processes of sample preparation for biological analysis. In this paper, the recent advancements of magnetic separation techniques based on magnetic materials in the field of sample preparation for biological analysis were reviewed. The strategy of magnetic separation techniques was summarized. The synthesis, stabilization and bio-functionalization of magnetic nanoparticles were reviewed in detail. Characterization of magnetic materials was also summarized. Moreover, the applications of magnetic separation techniques for the enrichment of protein, nucleic acid, cell, bioactive compound and immobilization of enzyme were described. Finally, the existed problems and possible trends of magnetic separation techniques for biological analysis in the future were proposed.

  20. Fluidics platform and method for sample preparation and analysis

    SciTech Connect

    Benner, W. Henry; Dzenitis, John M.; Bennet, William J.; Baker, Brian R.

    2014-08-19

    Herein provided are fluidics platform and method for sample preparation and analysis. The fluidics platform is capable of analyzing DNA from blood samples using amplification assays such as polymerase-chain-reaction assays and loop-mediated-isothermal-amplification assays. The fluidics platform can also be used for other types of assays and analyzes. In some embodiments, a sample in a sealed tube can be inserted directly. The following isolation, detection, and analyzes can be performed without a user's intervention. The disclosed platform may also comprises a sample preparation system with a magnetic actuator, a heater, and an air-drying mechanism, and fluid manipulation processes for extraction, washing, elution, assay assembly, assay detection, and cleaning after reactions and between samples.

  1. Method and apparatus for the preparation of liquid samples for determination of boron

    DOEpatents

    Siemer, Darryl D.

    1986-03-04

    A method and apparatus for the preparation of a liquid sample for the quantitative determination of boron by flame photometry. The sample is combined in a vessel with sulfuric acid, and an excess of methanol is added thereto. The methanol reacts with any boron present in the sample to form trimethyl borate which is volatilized by the heat of reaction between the excess methanol and sulfuric acid. The volatilized trimethyl borate is withdrawn from the vessel by either a partial vacuum or a positive pressure and is rapidly transferred to a standard flame photometer. The method is free of interference from typical boron concomitants.

  2. Method and apparatus for the preparation of liquid samples for determination of boron

    DOEpatents

    Siemer, Darryl D.

    1986-01-01

    A method and apparatus for the preparation of a liquid sample for the quantitative determination of boron by flame photometry. The sample is combined in a vessel with sulfuric acid, and an excess of methanol is added thereto. The methanol reacts with any boron present in the sample to form trimethyl borate which is volatilized by the heat of reaction between the excess methanol and sulfuric acid. The volatilized trimethyl borate is withdrawn from the vessel by either a partial vacuum or a positive pressure and is rapidly transferred to a standard flame photometer. The method is free of interference from typical boron concomitants.

  3. Method and apparatus for the preparation of liquid samples for determination of boron

    DOEpatents

    Siemer, D.D.

    A method and apparatus are described for the preparation of a liquid sample for the quantitative determination of boron by flame photometry. The sample is combined in a vessel with sulfuric acid, and an excess of methanol is added thereto. The methanol reacts with any boron present in the sample to form trimethyl borate which is volatilized by the heat of reaction between the excess methanol and sulfuric acid. The volatilized trimethyl borate is withdrawn from the vessel by either a partial vacuum or a positive pressure and is rapidly transferred to a standard flame photometer. The method is free of interference from typical boron concomitants.

  4. Enhanced Resolution of Chiral Amino Acids with Capillary Electrophoresis for Biosignature Detection in Extraterrestrial Samples.

    PubMed

    Creamer, Jessica S; Mora, Maria F; Willis, Peter A

    2017-01-17

    Amino acids are fundamental building blocks of terrestrial life as well as ubiquitous byproducts of abiotic reactions. In order to distinguish between amino acids formed by abiotic versus biotic processes it is possible to use chemical distributions to identify patterns unique to life. This article describes two capillary electrophoresis methods capable of resolving 17 amino acids found in high abundance in both biotic and abiotic samples (seven enantiomer pairs d/l-Ala, -Asp, -Glu, -His, -Leu, -Ser, -Val and the three achiral amino acids Gly, β-Ala, and GABA). To resolve the 13 neutral amino acids one method utilizes a background electrolyte containing γ-cyclodextrin and sodium taurocholate micelles. The acidic amino acid enantiomers were resolved with γ-cyclodextrin alone. These methods allow detection limits down to 5 nM for the neutral amino acids and 500 nM for acidic amino acids and were used to analyze samples collected from Mono Lake with minimal sample preparation.

  5. New Methods of Sample Preparation for Atom Probe Specimens

    NASA Technical Reports Server (NTRS)

    Kuhlman, Kimberly, R.; Kowalczyk, Robert S.; Ward, Jennifer R.; Wishard, James L.; Martens, Richard L.; Kelly, Thomas F.

    2003-01-01

    Magnetite is a common conductive mineral found on Earth and Mars. Disk-shaped precipitates approximately 40 nm in diameter have been shown to have manganese and aluminum concentrations. Atom-probe field-ion microscopy (APFIM) is the only technique that can potentially quantify the composition of these precipitates. APFIM will be used to characterize geological and planetary materials, analyze samples of interest for geomicrobiology; and, for the metrology of nanoscale instrumentation. Prior to APFIM sample preparation was conducted by electropolishing, the method of sharp shards (MSS), or Bosch process (deep reactive ion etching) with focused ion beam (FIB) milling as a final step. However, new methods are required for difficult samples. Many materials are not easily fabricated using electropolishing, MSS, or the Bosch process, FIB milling is slow and expensive, and wet chemistry and the reactive ion etching are typically limited to Si and other semiconductors. APFIM sample preparation using the dicing saw is commonly used to section semiconductor wafers into individual devices following manufacture. The dicing saw is a time-effective method for preparing high aspect ratio posts of poorly conducting materials. Femtosecond laser micromachining is also suitable for preparation of posts. FIB time required is reduced by about a factor of 10 and multi-tip specimens can easily be fabricated using the dicing saw.

  6. Process for the preparation of lactic acid and glyceric acid

    DOEpatents

    Jackson, James E [Haslett, MI; Miller, Dennis J [Okemos, MI; Marincean, Simona [Dewitt, MI

    2008-12-02

    Hexose and pentose monosaccharides are degraded to lactic acid and glyceric acid in an aqueous solution in the presence of an excess of a strongly anionic exchange resin, such as AMBERLITE IRN78 and AMBERLITE IRA400. The glyceric acid and lactic acid can be separated from the aqueous solution. Lactic acid and glyceric acid are staple articles of commerce.

  7. Preparation of water samples for carbon-14 dating

    USGS Publications Warehouse

    Feltz, H.R.; Hanshaw, Bruce B.

    1963-01-01

    For most natural water, a large sample is required to provide the 3 grams of carbon needed for a carbon-14 determination. A field procedure for isolating total dissolved-carbonate species is described. Carbon dioxide gas is evolved by adding sulfuric acid to the water sample; the gas is then collected in a sodium hydroxide trap by recycling in a closed system. The trap is then transported to the dating laboratory where the carbon-14 is counted.

  8. Alternate method of source preparation for alpha spectrometry: No electrodeposition, no hydrofluoric acid

    DOE PAGES

    Kurosaki, Hiromu; Mueller, Rebecca J.; Lambert, Susan B.; ...

    2016-07-15

    An alternate method of preparing actinide alpha counting sources was developed in place of electrodeposition or lanthanide fluoride micro-precipitation. The method uses lanthanide hydroxide micro-precipitation to avoid the use of hazardous hydrofluoric acid. Lastly, it provides a quicker, simpler, and safer way of preparing actinide alpha counting sources in routine, production-type laboratories that process many samples daily.

  9. Sample treatment and preparation for laser-induced breakdown spectroscopy

    NASA Astrophysics Data System (ADS)

    Jantzi, Sarah C.; Motto-Ros, Vincent; Trichard, Florian; Markushin, Yuri; Melikechi, Noureddine; De Giacomo, Alessandro

    2016-01-01

    One of the most widely cited advantages of laser-induced breakdown spectroscopy (LIBS) is that it does not require sample preparation, but this may also be the biggest factor holding it back from becoming a mature analytical technique like LA-ICP-MS, ICP-OES, or XRF. While there are certain specimen types that have enjoyed excellent LIBS results without any sample treatment (mostly homogeneous solids such as metals, glass, and polymers), the possible applications of LIBS have been greatly expanded through the use of sample preparation techniques that have resulted in analytical performance (i.e., limits of detection, accuracy, and repeatability) on par with XRF, ICP-OES, and often ICP-MS. This review highlights the work of many LIBS researchers who have developed, adapted, and improved upon sample preparation techniques for various specimen types in order to improve the quality of the analytical data that LIBS can produce in a large number of research domains. Strategies, not only for solids, but also liquids, gases, and aerosols are discussed, including newly developed nanoparticle enhancement and biological imaging and tagging techniques.

  10. Quantitation of protein in samples prepared for 2-D electrophoresis.

    PubMed

    Berkelman, Tom

    2008-01-01

    The concentration of protein in a sample prepared for two dimensional (2-D) electrophoretic analysis is usually determined by protein assay. Reasons for this include the following. (1) Protein quantitation ensures that the amount of protein to be separated is appropriate for the gel size and visualization method. (2) Protein quantitation facilitates comparison among similar samples, as image-based analysis is simplified when equivalent quantities of proteins have been loaded on the gels to be compared. (3) Quantitation is necessary in cases where the protein sample is labeled with dye before separation (1,2). The labeling chemistry is affected by the dye to protein ratio so it is essential to know the protein concentration before setting up the labeling reaction.A primary consideration with quantitating protein in samples prepared for 2-D electrophoresis is interference by nonprotein substances that may be present in the sample. These samples generally contain chaotropic solubilizing agents, detergents, reductants, buffers or carrier ampholytes, all of which potentially interfere with protein quantitation. The most commonly used protein assays in proteomics research are colorimetric assays in which the presence of protein causes a color change that can be measured spectrophotometrically (3). All protein assays utilize standards, a dilution series of a known concentration of a known protein, to create a standard curve. Two methods will be considered that circumvent some of the problems associated with interfering substances and are well suited for samples prepared for 2-D electrophoresis. The first method (4.1.1) relies on a color change that occurs upon binding of a dye to protein and the second (4.1.2) relies on binding and reduction of cupric ion (Cu2+) ion to cuprous ion (Cu+) by proteins.

  11. Target preparation for milligram sized 14C samples and data evaluation for AMS measurements

    NASA Astrophysics Data System (ADS)

    Andree, Michael; Beer, Jürg; Oeschger, Hans; Bonani, G.; Hofmann, H. J.; Morenzoni, E.; Nessi, M.; Suter, M.; Wölfli, W.

    1984-11-01

    Our preparation technique produces in a glow-discharge an amorphous carbon deposit on a copper substrate. The process starts with 1.6 cm 3 CO 2 STP (900 μg carbon) which is reduced over hot zinc to CO and subsequently cracked in the discharge. The yield of the process is typically 80%. With these targets in the Zürich ion source ion currents up to 20 μA are obtained. The background of samples prepared with this technique is presently around 30 ka (2.5% MODERN). The precision after half an hour measuring time for a modern sample is 0.7% and 2.7% for a three half-lives old sample, including the errors of the background and the NBS oxalic acid measurement. The method we use to correct for the background of the preparation and the accelerator as well as for the fractionation in the accelerator is presented.

  12. Sample preparation methods for determination of drugs of abuse in hair samples: A review.

    PubMed

    Vogliardi, Susanna; Tucci, Marianna; Stocchero, Giulia; Ferrara, Santo Davide; Favretto, Donata

    2015-02-01

    Hair analysis has assumed increasing importance in the determination of substances of abuse, both in clinical and forensic toxicology investigations. Hair analysis offers particular advantages over other biological matrices (blood and urine), including a larger window of detection, ease of collection and sample stability. In the present work, an overview of sample preparation techniques for the determination of substances of abuse in hair is provided, specifically regarding the principal steps in hair sample treatment-decontamination, extraction and purification. For this purpose, a survey of publications found in the MEDLINE database from 2000 to date was conducted. The most widely consumed substances of abuse and psychotropic drugs were considered. Trends in simplification of hair sample preparation, washing procedures and cleanup methods are discussed. Alternative sample extraction techniques, such as head-space solid phase microextraction (HS-SPDE), supercritical fluid extraction (SFE) and molecularly imprinted polymers (MIP) are also reported.

  13. Investigation into Alternative Sample Preparation Techniques for the Determination of Heavy Metals in Stationary Source Emission Samples Collected on Quartz Filters

    PubMed Central

    Goddard, Sharon L.; Brown, Richard J. C.

    2014-01-01

    Monitoring stationary source emissions for heavy metals generally requires the use of quartz filters to collect samples because of the high temperature and high moisture sampling environment. The documentary standard method sample preparation technique in Europe, EN 14385, uses digestion in hydrofluoric acid and nitric acid (HF/HNO3) followed by complexing with boric acid (H3BO3) prior to analysis. However, the use of this method presents a number of problems, including significant instrumental drift during analysis caused by the matrix components, often leading to instrument breakdown and downtime for repairs, as well as posing significant health and safety risks. The aim of this work was to develop an alternative sample preparation technique for emissions samples on quartz filters. The alternative techniques considered were: (i) acid digestion in a fluoroboric acid (HBF4) and HNO3 mixture and (ii) acid extraction in an aqua regia (AR) mixture (HCl and HNO3). Assessment of the effectiveness of these options included determination of interferences and signal drift, as well as validating the different methods by measurement of matrix certified reference materials (CRMs), and comparing the results obtained from real test samples and sample blanks to determine limits of detection. The results showed that the HBF4/HNO3 mixture provides the most viable alternative to the documentary standard preparation technique. PMID:25407906

  14. Preparation of SELEX Samples for Next-Generation Sequencing.

    PubMed

    Tolle, Fabian; Mayer, Günter

    2016-01-01

    Fuelled by massive whole genome sequencing projects such as the human genome project, enormous technological advancements and therefore tremendous price drops could be achieved, rendering next-generation sequencing very attractive for deep sequencing of SELEX libraries. Herein we describe the preparation of SELEX samples for Illumina sequencing, based on the already established whole genome sequencing workflow. We describe the addition of barcode sequences for multiplexing and the adapter ligation, avoiding associated pitfalls.

  15. Sample preparation and detection device for infectious agents

    DOEpatents

    Miles, Robin R.; Wang, Amy W.; Fuller, Christopher K.; Lemoff, Asuncion V.; Bettencourt, Kerry A.; Yu, June

    2003-06-10

    A sample preparation and analysis device which incorporates both immunoassays and PCR assays in one compact, field-portable microchip. The device provides new capabilities in fluid and particle control which allows the building of a fluidic chip with no moving parts, thus decreasing fabrication cost and increasing the robustness of the device. The device can operate in a true continuous (not batch) mode. The device incorporates magnetohydrodynamic (MHD) pumps to move the fluid through the system, acoustic mixing and fractionation, dielectropheretic (DEP) sample concentration and purification, and on-chip optical detection capabilities.

  16. Drug discovery from Nature: automated high-quality sample preparation

    PubMed Central

    Thiericke, Ralf

    2000-01-01

    Secondary metabolites from plants, animals and microorganisms have been proven to be an outstanding source for new and innovative drugs and show a striking structural diversity that supplements chemically synthesized compounds or libraries in drug discovery programs. Unfortunately, extracts from natural sources are usually complex mixtures of compounds:: often generated in time consuming and for the most part manual processes. As quality and quantity of the provided samples play a pivotal role in the success of high-throughput screening programs this poses serious problems. In order to make samples of natural origin competitive with synthetic compound libraries, we devised a novel, automated sample preparation procedure based on solid-phase extraction (SPE). By making use of a modified Zymark RapidTrace® SPE workstation an easy-to-handle and effective fractionation method has been developed which allows the generation of highquality samples from natural origin, fulfilling the requirements of an integration into high-throughput screening programs. PMID:18924703

  17. Apparatus for preparing a sample for mass spectrometry

    DOEpatents

    Villa-Aleman, E.

    1994-05-10

    An apparatus is described for preparing a sample for analysis by a mass spectrometer system. The apparatus has an entry chamber and an ionization chamber separated by a skimmer. A capacitor having two space-apart electrodes followed by one or more ion-imaging lenses is disposed in the ionization chamber. The chamber is evacuated and the capacitor is charged. A valve injects a sample gas in the form of sample pulses into the entry chamber. The pulse is collimated by the skimmer and enters the ionization chamber. When the sample pulse passes through the gap between the electrodes, it discharges the capacitor and is thereby ionized. The ions are focused by the imaging lenses and enter the mass analyzer, where their mass and charge are analyzed. 1 figures.

  18. Apparatus for preparing a sample for mass spectrometry

    DOEpatents

    Villa-Aleman, Eliel

    1994-01-01

    An apparatus for preparing a sample for analysis by a mass spectrometer system. The apparatus has an entry chamber and an ionization chamber separated by a skimmer. A capacitor having two space-apart electrodes followed by one or more ion-imaging lenses is disposed in the ionization chamber. The chamber is evacuated and the capacitor is charged. A valve injects a sample gas in the form of sample pulses into the entry chamber. The pulse is collimated by the skimmer and enters the ionization chamber. When the sample pulse passes through the gap between the electrodes, it discharges the capacitor and is thereby ionized. The ions are focused by the imaging lenses and enter the mass analyzer, where their mass and charge are analyzed.

  19. Recent advances in sample preparation techniques for effective bioanalytical methods.

    PubMed

    Kole, Prashant Laxman; Venkatesh, Gantala; Kotecha, Jignesh; Sheshala, Ravi

    2011-01-01

    This paper reviews the recent developments in bioanalysis sample preparation techniques and gives an update on basic principles, theory, applications and possibilities for automation, and a comparative discussion on the advantages and limitation of each technique. Conventional liquid-liquid extraction (LLE), protein precipitation (PP) and solid-phase extraction (SPE) techniques are now been considered as methods of the past. The last decade has witnessed a rapid development of novel sample preparation techniques in bioanalysis. Developments in SPE techniques such as selective sorbents and in the overall approach to SPE, such as hybrid SPE and molecularly imprinted polymer SPE, have been addressed. Considerable literature has been published in the area of solid-phase micro-extraction and its different versions, e.g. stir bar sorptive extraction, and their application in the development of selective and sensitive bioanalytical methods. Techniques such as dispersive solid-phase extraction, disposable pipette extraction and micro-extraction by packed sorbent offer a variety of extraction phases and provide unique advantages to bioanalytical methods. On-line SPE utilizing column-switching techniques is rapidly gaining acceptance in bioanalytical applications. PP sample preparation techniques such as PP filter plates/tubes offer many advantages like removal of phospholipids and proteins in plasma/serum. Newer approaches to conventional LLE techniques (salting-out LLE) are also covered in this review article.

  20. Sampling of illicit drugs for quantitative analysis--part III: sampling plans and sample preparations.

    PubMed

    Csesztregi, T; Bovens, M; Dujourdy, L; Franc, A; Nagy, J

    2014-08-01

    The findings in this paper are based on the results of our drug homogeneity studies and particle size investigations. Using that information, a general sampling plan (depicted in the form of a flow-chart) was devised that could be applied to the quantitative instrumental analysis of the most common illicit drugs: namely heroin, cocaine, amphetamine, cannabis resin, MDMA tablets and herbal cannabis in 'bud' form (type I). Other more heterogeneous forms of cannabis (type II) were found to require alternative, more traditional sampling methods. A table was constructed which shows the sampling uncertainty expected when a particular number of random increments are taken and combined to form a single primary sample. It also includes a recommended increment size; which is 1 g for powdered drugs and cannabis resin, 1 tablet for MDMA and 1 bud for herbal cannabis in bud form (type I). By referring to that table, individual laboratories can ensure that the sampling uncertainty for a particular drug seizure can be minimised, such that it lies in the same region as their analytical uncertainty for that drug. The table shows that assuming a laboratory wishes to quantitatively analyse a seizure of powdered drug or cannabis resin with a 'typical' heterogeneity, a primary sample of 15×1 g increments is generally appropriate. The appropriate primary sample for MDMA tablets is 20 tablets, while for herbal cannabis (in bud form) 50 buds were found to be appropriate. Our study also showed that, for a suitably homogenised primary sample of the most common powdered drugs, an analytical sample size of between 20 and 35 mg was appropriate and for herbal cannabis the appropriate amount was 200 mg. The need to ensure that the results from duplicate or multiple incremental sampling were compared, to demonstrate whether or not a particular seized material has a 'typical' heterogeneity and that the sampling procedure applied has resulted in a 'correct sample', was highlighted and the setting

  1. Optimization for Peptide Sample Preparation for Urine Peptidomics

    SciTech Connect

    Sigdel, Tara K.; Nicora, Carrie D.; Hsieh, Szu-Chuan; Dai, Hong; Qian, Weijun; Camp, David G.; Sarwal, Minnie M.

    2014-02-25

    Analysis of native or endogenous peptides in biofluids can provide valuable insights into disease mechanisms. Furthermore, the detected peptides may also have utility as potential biomarkers for non-invasive monitoring of human diseases. The non-invasive nature of urine collection and the abundance of peptides in the urine makes analysis by high-throughput ‘peptidomics’ methods , an attractive approach for investigating the pathogenesis of renal disease. However, urine peptidomics methodologies can be problematic with regards to difficulties associated with sample preparation. The urine matrix can provide significant background interference in making the analytical measurements that it hampers both the identification of peptides and the depth of the peptidomics read when utilizing LC-MS based peptidome analysis. We report on a novel adaptation of the standard solid phase extraction (SPE) method to a modified SPE (mSPE) approach for improved peptide yield and analysis sensitivity with LC-MS based peptidomics in terms of time, cost, clogging of the LC-MS column, peptide yield, peptide quality, and number of peptides identified by each method. Expense and time requirements were comparable for both SPE and mSPE, but more interfering contaminants from the urine matrix were evident in the SPE preparations (e.g., clogging of the LC-MS columns, yellowish background coloration of prepared samples due to retained urobilin, lower peptide yields) when compared to the mSPE method. When we compared data from technical replicates of 4 runs, the mSPE method provided significantly improved efficiencies for the preparation of samples from urine (e.g., mSPE peptide identification 82% versus 18% with SPE; p = 8.92E-05). Additionally, peptide identifications, when applying the mSPE method, highlighted the biology of differential activation of urine peptidases during acute renal transplant rejection with distinct laddering of specific peptides, which was obscured for most proteins

  2. Properties of Copolymers of Aspartic Acid and Aliphatic Dicarboxylic Acids Prepared by Reactive Extrusion

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspartic acid may be prepared chemically or by the fermentation of carbohydrates. Currently, low molecular weight polyaspartic acids are prepared commercially by heating aspartic acid at high temperatures (greater than 220 degrees C) for several hours in the solid state. In an effort to develop a ...

  3. Combining Electrochemical Sensors with Miniaturized Sample Preparation for Rapid Detection in Clinical Samples

    PubMed Central

    Bunyakul, Natinan; Baeumner, Antje J.

    2015-01-01

    Clinical analyses benefit world-wide from rapid and reliable diagnostics tests. New tests are sought with greatest demand not only for new analytes, but also to reduce costs, complexity and lengthy analysis times of current techniques. Among the myriad of possibilities available today to develop new test systems, amperometric biosensors are prominent players—best represented by the ubiquitous amperometric-based glucose sensors. Electrochemical approaches in general require little and often enough only simple hardware components, are rugged and yet provide low limits of detection. They thus offer many of the desirable attributes for point-of-care/point-of-need tests. This review focuses on investigating the important integration of sample preparation with (primarily electrochemical) biosensors. Sample clean up requirements, miniaturized sample preparation strategies, and their potential integration with sensors will be discussed, focusing on clinical sample analyses. PMID:25558994

  4. Effects of sample preparation on bacterial colonization of polymers.

    PubMed

    Zeiger, Diana N; Stafford, Christopher M; Cheng, Yajun; Leigh, Stefan D; Lin-Gibson, Sheng; Lin, Nancy J

    2010-02-16

    Characterization of materials developed for medical usage frequently includes studies in which the materials are inoculated with bacteria in order to assess bacterial colonization and biofilm formation. Observed differences in bacterial growth are typically considered to be due to the material or the incubation conditions. To our knowledge, the method used to prepare the materials has generally not been considered with regard to its influence on bacterial colonization. The objective of this study was to determine the effects that various preparation methods exert on bacterial colonization of polymer disks. Polymer disks of the same dimethacrylate composition were photopolymerized: (1) between untreated glass slides, (2) between polyester release film, (3) between glass slides treated with an alkyl silane, (4) between glass slides treated with a perfluorinated silane, or (5) with one free surface in an argon-purged chamber. Surface chemistry was quantified using X-ray photoelectron spectroscopy, hydrophobicity was assessed by water contact angle, and topography was characterized using atomic force microscopy. The disks were inoculated with Streptococcus mutans for 4 h, fixed, and visualized using confocal laser scanning microscopy. Differences among all groups were found with regard to surface chemistry, hydrophobicity, topography, and bacteria morphology, density, and coverage, indicating that the method of sample preparation strongly affects both the surface properties and the initial bacterial colonization. Polymerization on untreated slides was selected as the preferred method of preparation due to minimal material transfer to the polymer and consistent, reproducible bacterial colonization.

  5. Preparation of DNA and nucleoprotein samples for AFM imaging

    PubMed Central

    Lyubchenko, Yuri L.

    2010-01-01

    Sample preparation techniques allowing reliable and reproducible imaging of DNA with various structures, topologies and complexes with proteins are reviewed. The major emphasis is given to methods utilizing chemical functionalization of mica, enabling preparation of the surfaces with required characteristics. The methods are illustrated by examples of imaging of different DNA structures. Special attention is given to the possibility of AFM to image the dynamics of DNA at the nanoscale. The capabilities of time-lapse AFM in aqueous solutions are illustrated by imaging of dynamic processes as transitions of local alternative structures (transition of DNA between H and B forms). The application of AFM to studies of protein-DNA complexes is illustrated by a few examples of imaging site-specific complexes, as well as such systems as chromatin. The time-lapse AFM studies of protein-DNA complexes including very recent advances with the use of high-speed AFM are reviewed. PMID:20864349

  6. Isotopic analysis of eggs: evaluating sample collection and preparation.

    PubMed

    Rock, Luc; Rowe, Sylwia; Czerwiec, Agnieszka; Richmond, Harold

    2013-02-15

    Egg traceability/authenticity is a worldwide concern. Stable isotope techniques have been suggested as a tool to address this issue. To further validate the use of these techniques, a research project was undertaken to evaluate what effect sample collection and preparation have on the measured isotopic composition of egg components. The timing of egg collection, the timing of egg preparation after collection, and the use of pasteurisation were investigated. The C, N, O, and S isotopic compositions of egg components from 7 different production systems were measured. Two sets of eggs were collected (4 months apart). It was found that the 'isotopic fingerprint' of a particular production system was maintained over time, and that it may be possible to trace liquid egg products based on isotopic data from fresh eggs. The findings from this study support the integration of stable isotope techniques in egg traceability/authenticity systems.

  7. Leaching of metals from steel samples in peracetic acid

    NASA Astrophysics Data System (ADS)

    Yabutani, Tomoki; Nakamura, Takamasa; Takayabagi, Toshio

    2015-03-01

    In this paper, leaching behavior of metallic species from steel samples in peracetic acid was investigated. We compared the leaching efficiency between peracetic acid and acetic acid to estimate the role of peroxo functional group for the leaching. As a result, peracetic acid enhanced the leaching ability of metallic species from the high speed steel and the alloy steel samples. MoO3, Mo, MO2C, W, WO3, VC and MnO2 were effectively leached by peracetic acid, while the stainless steel had a high resistance against corrosion by peracetic acid.

  8. 40 CFR 1065.1107 - Sample media and sample system preparation; sample system assembly.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... number of repeat tests. Note that when repeating test cycles to increase sample mass, you may replace the... with methylene chloride for 24 hours and dry over dry nitrogen in a low-temperature vacuum oven. (2... methanol, 22 hours with methylene chloride, and 22 hours with toluene, followed by drying with nitrogen....

  9. Sampling and sample preparation methods for determining concentrations of mycotoxins in foods and feeds.

    PubMed

    2012-01-01

    Sample variation is often the largest error in determining concentrations of mycotoxins in food commodities. The worldwide safety evaluation of mycotoxins requires sampling plans that give acceptably accurate values for the levels of contamination in specific batches or lots of a commodity. Mycotoxin concentrations show a skewed or uneven distribution in foods and feeds, especially in whole kernels (or nuts), so it is extremely difficult to collect a sample that accurately represents the mean batch concentration. Sample variance studies and sampling plans have been published for select mycotoxins such as aflatoxin, fumonisin, and deoxynivalenol, emphasizing the importance of sample selection, sample size, and the number of incremental samples. For meaningful data to be generated from surveillance studies, representative samples should be collected from carefully selected populations (batches or lots) of food that, in turn, should be representative of clearly defined locations (e.g. a country, a region within a country). Although sampling variability is unavoidable, it is essential that the precision of the sampling plan be clearly defined and be considered acceptable by those responsible for interpreting and reporting the surveillance data. The factors influencing variability are detailed here, with reference to both major mycotoxins and major commodities. Sampling of large bag stacks, bulk shipments, and domestic supplies are all discussed. Sampling plans currently accepted in international trade are outlined. Acceptance sampling plans and the variabilities that affect operating characteristic curves of such plans are also detailed. The constraints and issues related to the sampling of harvested crops within subsistence farming areas are also discussed in this chapter, as are the essential rules of sample labelling and storage. The chapter concludes with a short section on sample preparation methods.

  10. Mixing apparatus for preparing NMR samples under pressure

    NASA Astrophysics Data System (ADS)

    Wu, Wen-Jin; Vidugiris, Gediminas; Mooberry, Ed S.; Westler, William M.; Markley, John L.

    2003-09-01

    The size limit for protein NMR spectroscopy in solution arises in large part from line broadening caused by slow molecular tumbling. One way to alleviate this problem is to increase the effective tumbling rate by reducing the viscosity of the solvent. Because proteins generally require an aqueous environment to remain folded, one approach has been to encapsulate hydrated proteins in reverse micelles formed by a detergent and to dissolve the encapsulated protein in a low-viscosity fluid. The high volatility of suitable low-viscosity fluids requires that the samples be prepared and maintained under pressure. We describe a novel apparatus used for the preparation of such samples. The apparatus includes a chamber for mixing the detergent with the low-viscosity solvent, a second chamber for mixing this with hydrated protein, and a 5-mm (o.d.) zirconium oxide NMR sample tube with shut-off valves designed to contain pressures on the order of 10 bar, sufficient for liquid propane. Liquids are moved from one location to another by introducing minor pressure differentials between two pressurization vessels. We discuss the operation of this apparatus and illustrate this with data on a 30-kDa protein complex (chymotrypsin:turkey ovomucoid third domain) encapsulated in reverse micelles of the detergent, sodium bis (2-ethylhexyl) sulfosuccinate, aerosol-ot (AOT), dissolved in liquid propane.

  11. Sample preparation and EFTEM of Meat Samples for Nanoparticle Analysis in Food

    NASA Astrophysics Data System (ADS)

    Lari, L.; Dudkiewicz, A.

    2014-06-01

    Nanoparticles are used in industry for personal care products and the preparation of food. In the latter application, their functions include the prevention of microbes' growth, increase of the foods nutritional value and sensory quality. EU regulations require a risk assessment of the nanoparticles used in foods and food contact materials before the products can reach the market. However, availability of validated analytical methodologies for detection and characterisation of the nanoparticles in food hampers appropriate risk assessment. As part of a research on the evaluation of the methods for screening and quantification of Ag nanoparticles in meat we have tested a new TEM sample preparation alternative to resin embedding and cryo-sectioning. Energy filtered TEM analysis was applied to evaluate thickness and the uniformity of thin meat layers acquired at increasing input of the sample demonstrating that the protocols used ensured good stability under the electron beam, reliable sample concentration and reproducibility.

  12. Comparison of sample preparation methods for the recovery of foodborne pathogens from fresh produce.

    PubMed

    Kim, Se-Ri; Yoon, Yohan; Kim, Won-Il; Park, Kyeong-Hun; Yun, Hye-Jeong; Chung, Duck Hwa; Yun, Jong Chul; Ryu, Kyoung Yul

    2012-07-01

    Sample preparation methods (pummeling, pulsifying, sonication, and shaking by hand) were compared for achieving maximum recovery of foodborne pathogens from iceberg lettuce, perilla leaves, cucumber, green pepper, and cherry tomato. Antimicrobial and dehydration effects also were examined to investigate causes of poor recovery of pathogens. Each produce type was inoculated with Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus at 6.0 log CFU/cm(2), and samples were prepared using the four methods. Bacterial populations recovered from the five types of produce were significantly different (P < 0.05) according to sample preparation methods and produce type. The bacterial populations recovered from pummeled and pulsified samples were higher (P < 0.05) than those recovered from sonicated and hand-shaken samples, except for cherry tomato. The number of bacteria recovered from produce was reduced (P < 0.05) from that of the inoculum by 0.16 to 2.69 log CFU/cm(2). Although extracts of iceberg lettuce, perilla leaves, cucumber, and green pepper had no antimicrobial activity, the populations of E. coli O157:H7, Salmonella Typhimurium, B. cereus, and L. monocytogenes in cherry tomato extract were slightly reduced after these treatments (P < 0.05). The pathogen populations on perilla leaves and cherry tomatoes decreased by >2 log CFU/cm(2) after exposure to 40% relative humidity for 1 h. No reduction was observed when the five pathogens were exposed to 90% relative humidity. These data suggest that pummeling and pulsifying are optimal sample preparation methods for detection of microorganisms. Acidic produce such as cherry tomato should be treated with a method that does not cause sample breakdown so that acid stress on the bacteria can be minimized. Dehydration stress also affects recovery of pathogens from produce.

  13. Distribution and Origin of Amino Acids in Lunar Regolith Samples

    NASA Technical Reports Server (NTRS)

    Elsila, J. E.; Callahan, M. P.; Glavin, D. P.; Dworkin, J. P.; McLain, H. L.; Noble, S. K.; Gibson, E. K., Jr.

    2015-01-01

    The existence of organic compounds on the lunar surface has been a question of interest from the Apollo era to the present. Investigations of amino acids immediately after collection of lunar samples yielded inconclusive identifications, in part due to analytical limitations including insensitivity to certain compounds, an inability to separate enantiomers, and lack of compound-specific isotopic measurements. It was not possible to determine if the detected amino acids were indigenous to the lunar samples or the result of terrestrial contamination. Recently, we presented initial data from the analysis of amino acid abundances in 12 lunar regolith samples and discussed those results in the context of four potential amino acid sources [5]. Here, we expand on our previous work, focusing on amino acid abundances and distributions in seven regolith samples and presenting the first compound-specific carbon isotopic ratios measured for amino acids in a lunar sample.

  14. An apparatus for preparing benthic samples aboard ship

    USGS Publications Warehouse

    Pepper, Phillip N.; Girard, Thomas L.; Stapanian, Martin A.

    2001-01-01

    We describe a safe and effective apparatus for washing and reducing the volume of benthic samples collected by grab samplers aboard ship. The sample is transferred directly from the dredge to the apparatus and then washed with water pumped through pipes in the apparatus and from onboard hoses. Wastewater and materials smaller than 0.541 mm in diameter are washed overboard. Larger materials, including benthic organisms, collect on an upper 0.64-cm screen and on a lower 30-mm-mesh stainless steel bolt cloth. A collection jar is screwed into the bottom of the apparatus. Therefore, transfer of sample material from the apparatus to the jar is quick and easy. This apparatus has several advantages for use aboard ship over others described in the literature, especially in rough seas, in cold weather, and at night. The apparatus provides a safe and convenient platform for washing and reducing samples, and samples can be prepared while the vessel is traveling at full speed.

  15. On sample preparation and dielectric breakdown in nanostructured epoxy resins

    NASA Astrophysics Data System (ADS)

    Reading, M.; Xu, Z.; Vaughan, A. S.; Lewin, P. L.

    2011-08-01

    There are many methods available to achieve a good dispersion of fillers within a polymeric matrix. This investigation considered several methods of dispersing three chosen fillers within an epoxy resin; the same processes were also performed on unfilled materials to investigate any effects they may have on the host material. For this investigation, the epoxy system (EP) was combined with sodium montmorillonite (MMT), micrometric silicon dioxide (SD) or nanometric silicon dioxide (NSD) as fillers. The effect of the different sample preparation routes on breakdown behaviour was then evaluated. While more thorough mixing protocols were found to lead to improved breakdown behaviour in the case of the various filled systems, surprisingly, an entirely equivalent form of behaviour was also seen in the unfilled epoxy. The influence of changes in sample geometry on the breakdown strength was established.

  16. Study of exhaled breath condensate sample preparation for metabolomics analysis by LC-MS/MS in high resolution mode.

    PubMed

    Fernández-Peralbo, M A; Calderón Santiago, M; Priego-Capote, F; Luque de Castro, M D

    2015-11-01

    Metabolomic analysis of exhaled breath condensate (EBC) requires an unavoidable sample preparation step because of the low concentration of its components, and potential cleanup for possible interferents. Sample preparation based on protein precipitation (PP), solid-phase extraction (SPE) by hydrophilic and lipophilic sorbents or lyophilization has demonstrated that the analytical sample from the last is largely the best because lyophilization allows reconstitution in a volume as small as required (preconcentration factors up to 80-times with respect to the original sample), thus doubling the number of detected compounds as compared with the other alternatives (47 versus 25). In addition, PP and/or SPE cleanup are unnecessary as no effect from the EBC components removed by these steps appears in the chromatograms. The total 49 EBC compounds tentatively identified and confirmed by MS/MS in this research include amino acids, fatty acids, fatty amides, fatty aldehydes, sphingoid bases, oxoanionic compounds, imidazoles, hydroxy acids and aliphatic acyclic acids.

  17. Porous monoliths for on-line sample preparation: A review.

    PubMed

    Masini, Jorge C; Svec, Frantisek

    2017-04-29

    This review aims at presenting the state of the art concerning monolithic materials for on-line sample preparation emphasizing solid-phase extraction, matrix exchange, and analyte conversion. Emphasis was given to organic and silica-based, as well as hybrid monoliths reported in the literature mostly after 2010. The first part of this review presents materials and strategies for enrichment of inorganic species in environmental and biological samples using mostly ICP-MS detectors. In the second part we focus on organic analytes, discussing the role of surface area of the polymer monoliths and density of adsorption sites for specific interactions, including incorporation of nanoparticles, metal organic frameworks, as well as the preparation of hybrid organic-silica monoliths to increase the surface area. Incorporation of ionic liquids to increase the number of types of interaction mechanisms available for retention is also discussed. Monoliths affording molecular recognition properties achieved by including boronate moieties for cis-diol recognition, as well as antibodies and aptamers for specific molecular recognition are also reviewed. The largest number of applications of molecular recognition mechanisms was observed for molecularly imprinted polymer monoliths as a consequence of the simplicity of this approach when compared to the use of immunosorbents or aptamers. The final part examines the on-line applications of immobilized enzyme reactors used for protein digestion in proteomic analysis and for kinetic studies in drug discovery and clinical assays usually coupling the reactors to mass spectrometers.

  18. S- to N-Palmitoyl Transfer During Proteomic Sample Preparation

    NASA Astrophysics Data System (ADS)

    Ji, Yuhuan; Bachschmid, Markus M.; Costello, Catherine E.; Lin, Cheng

    2016-04-01

    N-palmitoylation has been reported in a number of proteins and suggested to play an important role in protein localization and functions. However, it remains unclear whether N-palmitoylation is a direct enzyme-catalyzed process, or results from intramolecular S- to N-palmitoyl transfer. Here, using the S-palmitoyl peptide standard, GCpalmLGNAK, as the model system, we observed palmitoyl migration from the cysteine residue to either the peptide N-terminus or the lysine side chain during incubation in both neutral and slightly basic buffers commonly used in proteomic sample preparation. Palmitoyl transfer can take place either intra- or inter-molecularly, with the peptide N-terminus being the preferred migration site, presumably because of its lower basicity. The extent of intramolecular palmitoyl migration was low in the system studied, as it required the formation of an entropically unfavored macrocycle intermediate. Intermolecular palmitoyl transfer, however, remained a tangible problem, and may lead to erroneous reporting of in vivo N-palmitoylation. It was found that addition of the MS-compatible detergent RapiGest could significantly inhibit intermolecular palmitoyl transfer, as well as thioester hydrolysis and DTT-induced thioester cleavage. Finally, palmitoyl transfer from the cysteine residue to the peptide N-terminus can also occur in the gas phase, during collision-induced dissociation, and result in false identification of N-palmitoylation. Therefore, one must be careful with both sample preparation and interpretation of tandem mass spectra in the study of N-palmitoylation.

  19. Preparation of .alpha.,.beta.-unsaturated carboxylic acids and esters

    DOEpatents

    Gogate, Makarand Ratnakar; Spivey, James Jerry; Zoeller, Joseph Robert

    1998-01-01

    Disclosed is a process for the preparation of .alpha.,.beta.-unsaturated carboxylic acids and esters thereof which comprises contacting formaldehyde or a source of formaldehyde with a carboxylic acid, ester or anhydride in the presence of a catalyst comprising an oxide of niobium.

  20. Preparation of {alpha},{beta}-unsaturated carboxylic acids and esters

    DOEpatents

    Gogate, M.R.; Spivey, J.J.; Zoeller, J.R.

    1998-09-15

    Disclosed is a process for the preparation of {alpha},{beta}-unsaturated carboxylic acids and esters thereof which comprises contacting formaldehyde or a source of formaldehyde with a carboxylic acid, ester or anhydride in the presence of a catalyst comprising an oxide of niobium.

  1. The Origin of Amino Acids in Lunar Regolith Samples

    NASA Technical Reports Server (NTRS)

    Cook, Jamie E.; Callahan, Michael P.; Dworkin, Jason P.; Glavin, Daniel P.; McLain, Hannah L.; Noble, Sarah K.; Gibson, Everett K., Jr.

    2016-01-01

    We analyzed the amino acid content of seven lunar regolith samples returned by the Apollo 16 and Apollo 17 missions and stored under NASA curation since collection using ultrahigh-performance liquid chromatography with fluorescence detection and time-of-flight mass spectrometry. Consistent with results from initial analyses shortly after collection in the 1970s, we observed amino acids at low concentrations in all of the curated samples, ranging from 0.2 parts-per-billion (ppb) to 42.7 ppb in hot-water extracts and 14.5 ppb to 651.1 ppb in 6M HCl acid-vapor-hydrolyzed, hot-water extracts. Amino acids identified in the Apollo soil extracts include glycine, D- and L-alanine, D- and L-aspartic acid, D- and L-glutamic acid, D- and L-serine, L-threonine, and L-valine, all of which had previously been detected in lunar samples, as well as several compounds not previously identified in lunar regoliths: -aminoisobutyric acid (AIB), D-and L-amino-n-butyric acid (-ABA), DL-amino-n-butyric acid, -amino-n-butyric acid, -alanine, and -amino-n-caproic acid. We observed an excess of the L enantiomer in most of the detected proteinogenic amino acids, but racemic alanine and racemic -ABA were present in some samples.

  2. Preparation of Soybean Seed Samples for FT-IR Microspectroscopy

    SciTech Connect

    Miller,S.; Pietrzak, L.

    2005-01-01

    Typical preparation of seed samples for infrared (IR) microspectroscopy involves imbibition of the seed for varying time periods followed by cryosectioning. Imbibition, however, may initiate germination even at 4 C with associated changes in the chemistry of the sample. We have found that it is possible to section seeds that are sufficiently hard, such as soybeans, on a standard laboratory microtome without imbibition. The use of dry sectioning of unimbibed seeds is reported here, as well as a comparison of different mounting media and modes of analysis. Glycerol, Tissue-Tek, and ethanol were used as mounting media, and the quality of the resulting spectra was assessed. Ethanol was the preferred mountant, because it dried quickly with no residue and thus did not interfere with the spectrum of interest. Analysis in transmission mode using barium fluoride windows to hold the samples was compared with transmission-reflection analysis with sections mounted on special infrared-reflecting slides. The two modes of analysis performed well in different regions of the spectrum. The mode of analysis (transmission vs. transmission-reflection) should be based on the components of greatest interest in the sample.

  3. Sample preparation of metal alloys by electric discharge machining

    NASA Technical Reports Server (NTRS)

    Chapman, G. B., II; Gordon, W. A.

    1976-01-01

    Electric discharge machining was investigated as a noncontaminating method of comminuting alloys for subsequent chemical analysis. Particulate dispersions in water were produced from bulk alloys at a rate of about 5 mg/min by using a commercially available machining instrument. The utility of this approach was demonstrated by results obtained when acidified dispersions were substituted for true acid solutions in an established spectrochemical method. The analysis results were not significantly different for the two sample forms. Particle size measurements and preliminary results from other spectrochemical methods which require direct aspiration of liquid into flame or plasma sources are reported.

  4. A self-contained polymeric cartridge for automated biological sample preparation.

    PubMed

    Xu, Guolin; Lee, Daniel Yoke San; Xie, Hong; Chiew, Deon; Hsieh, Tseng-Ming; Ali, Emril Mohamed; Lun Looi, Xing; Li, Mo-Huang; Ying, Jackie Y

    2011-09-01

    Sample preparation is one of the most crucial processes for nucleic acids based disease diagnosis. Several steps are required for nucleic acids extraction, impurity washes, and DNA/RNA elution. Careful sample preparation is vital to the obtaining of reliable diagnosis, especially with low copies of pathogens and cells. This paper describes a low-cost, disposable lab cartridge for automatic sample preparation, which is capable of handling flexible sample volumes of 10 μl to 1 ml. This plastic cartridge contains all the necessary reagents for pathogen and cell lysis, DNA/RNA extraction, impurity washes, DNA/RNA elution and waste processing in a completely sealed cartridge. The entire sample preparation processes are automatically conducted within the cartridge on a desktop unit using a pneumatic fluid manipulation approach. Reagents transportation is achieved with a combination of push and pull forces (with compressed air and vacuum, respectively), which are connected to the pneumatic inlets at the bottom of the cartridge. These pneumatic forces are regulated by pinch valve manifold and two pneumatic syringe pumps within the desktop unit. The performance of this pneumatic reagent delivery method was examined. We have demonstrated the capability of the on-cartridge RNA extraction and cancer-specific gene amplification from 10 copies of MCF-7 breast cancer cells. The on-cartridge DNA recovery efficiency was 54-63%, which was comparable to or better than the conventional manual approach using silica spin column. The lab cartridge would be suitable for integration with lab-chip real-time polymerase chain reaction devices in providing a portable system for decentralized disease diagnosis.

  5. Rapid Filtration Separation-Based Sample Preparation Method for Bacillus Spores in Powdery and Environmental Matrices

    PubMed Central

    Isabel, Sandra; Boissinot, Maurice; Charlebois, Isabelle; Fauvel, Chantal M.; Shi, Lu-E; Lévesque, Julie-Christine; Paquin, Amélie T.; Bastien, Martine; Stewart, Gale; Leblanc, Éric; Sato, Sachiko

    2012-01-01

    Authorities frequently need to analyze suspicious powders and other samples for biothreat agents in order to assess environmental safety. Numerous nucleic acid detection technologies have been developed to detect and identify biowarfare agents in a timely fashion. The extraction of microbial nucleic acids from a wide variety of powdery and environmental samples to obtain a quality level adequate for these technologies still remains a technical challenge. We aimed to develop a rapid and versatile method of separating bacteria from these samples and then extracting their microbial DNA. Bacillus atrophaeus subsp. globigii was used as a simulant of Bacillus anthracis. We studied the effects of a broad variety of powdery and environmental samples on PCR detection and the steps required to alleviate their interference. With a benchmark DNA extraction procedure, 17 of the 23 samples investigated interfered with bacterial lysis and/or PCR-based detection. Therefore, we developed the dual-filter method for applied recovery of microbial particles from environmental and powdery samples (DARE). The DARE procedure allows the separation of bacteria from contaminating matrices that interfere with PCR detection. This procedure required only 2 min, while the DNA extraction process lasted 7 min, for a total of <10 min. This sample preparation procedure allowed the recovery of cleaned bacterial spores and relieved detection interference caused by a wide variety of samples. Our procedure was easily completed in a laboratory facility and is amenable to field application and automation. PMID:22210204

  6. Rapid filtration separation-based sample preparation method for Bacillus spores in powdery and environmental matrices.

    PubMed

    Isabel, Sandra; Boissinot, Maurice; Charlebois, Isabelle; Fauvel, Chantal M; Shi, Lu-E; Lévesque, Julie-Christine; Paquin, Amélie T; Bastien, Martine; Stewart, Gale; Leblanc, Eric; Sato, Sachiko; Bergeron, Michel G

    2012-03-01

    Authorities frequently need to analyze suspicious powders and other samples for biothreat agents in order to assess environmental safety. Numerous nucleic acid detection technologies have been developed to detect and identify biowarfare agents in a timely fashion. The extraction of microbial nucleic acids from a wide variety of powdery and environmental samples to obtain a quality level adequate for these technologies still remains a technical challenge. We aimed to develop a rapid and versatile method of separating bacteria from these samples and then extracting their microbial DNA. Bacillus atrophaeus subsp. globigii was used as a simulant of Bacillus anthracis. We studied the effects of a broad variety of powdery and environmental samples on PCR detection and the steps required to alleviate their interference. With a benchmark DNA extraction procedure, 17 of the 23 samples investigated interfered with bacterial lysis and/or PCR-based detection. Therefore, we developed the dual-filter method for applied recovery of microbial particles from environmental and powdery samples (DARE). The DARE procedure allows the separation of bacteria from contaminating matrices that interfere with PCR detection. This procedure required only 2 min, while the DNA extraction process lasted 7 min, for a total of <10 min. This sample preparation procedure allowed the recovery of cleaned bacterial spores and relieved detection interference caused by a wide variety of samples. Our procedure was easily completed in a laboratory facility and is amenable to field application and automation.

  7. Formation of an artifact of diclofenac during acidic extraction of environmental water samples.

    PubMed

    Reddersen, K; Heberer, Th

    2003-09-05

    Solid-phase extraction at an acidic pH is used as a common sample preparation method for analyzing residues of the analgesic drug diclofenac (2-[(2,6-dichlorophenyl)amino] benzeneacetic acid) in environmental water samples. This paper describes the matrix-dependent formation of an artifact of diclofenac during sample preparation resulting in an up to 40% underestimation of diclofenac concentrations especially in matrix-prone samples such as sewage effluents or surface water. The artifact most likely being formed during acidification of the sample was unequivocally identified as 1-(2,6-dichlorophenyl)indolin-2-one by capillary gas chromatography-mass spectrometry. To avoid an underestimation of the analytical results quantification of both diclofenac and its artifact is recommended.

  8. An instrument for automated purification of nucleic acids from contaminated forensic samples.

    PubMed

    Broemeling, David J; Pel, Joel; Gunn, Dylan C; Mai, Laura; Thompson, Jason D; Poon, Hiron; Marziali, Andre

    2008-02-01

    Forensic crime scene sample analysis, by its nature, often deals with samples in which there are low amounts of nucleic acids, on substrates that often lead to inhibition of subsequent enzymatic reactions such as PCR amplification for STR profiling. Common substrates include denim from blue jeans, which yields indigo dye as a PCR inhibitor, and soil, which yields humic substances as inhibitors. These inhibitors frequently co-extract with nucleic acids in standard column or bead-based preps, leading to frequent failure of STR profiling. We present a novel instrument for DNA purification of forensic samples that is capable of highly effective concentration of nucleic acids from soil particulates, fabric, and other complex samples including solid components. The novel concentration process, known as SCODA, is inherently selective for long charged polymers such as DNA, and therefore is able to effectively reject known contaminants. We present an automated sample preparation instrument based on this process, and preliminary results based on mock forensic samples.

  9. Preparation and characterization Al3+-bentonite Turen Malang for esterification fatty acid (palmitic acid, oleic acid and linoleic acid)

    NASA Astrophysics Data System (ADS)

    Abdulloh, Abdulloh; Aminah, Nanik Siti; Triyono, Mudasir, Trisunaryanti, Wega

    2016-03-01

    Catalyst preparation and characterization of Al3+-bentonite for esterification of palmitic acid, oleic acid and linoleic acid has been done. Al3+-bentonite catalyst was prepared from natural bentonite of Turen Malang through cation exchange reaction using AlCl3 solution. The catalysts obtained were characterized by XRD, XRF, pyridine-FTIR and surface area analyser using the BET method. Catalyst activity test of Al3+-bentonite for esterification reaction was done at 65°C using molar ratio of metanol-fatty acid of 30:1 and 0.25 g of Al3+-bentonite catalyst for the period of ½, 1, 2, 3, 4 and 5 hours. Based on the characterization results, the Al3+-bentonite Turen Malang catalyst has a d-spacing of 15.63 Ǻ, acid sites of Brönsted and Lewis respectively of 230.79 µmol/g and 99.39 µmol/g, surface area of 507.3 m2/g and the average of radius pore of 20.09 Å. GC-MS analysis results of the oil phase after esterification reaction showed the formation of biodiesel (FAME: Fatty acid methyl ester), namely methyl palmitate, methyl oleate and methyl linoleate. The number of conversions resulted in esterification reaction using Al3+-bentonite Turen Malang catalyst was 74.61%, 37.75%, and 20, 93% for the esterification of palmitic acid, oleic acid and linoleic acid respectively.

  10. The role of sample preparation in interpretation of trace element concentration variability in moss bioindication studies

    USGS Publications Warehouse

    Migaszewski, Z.M.; Lamothe, P.J.; Crock, J.G.; Galuszka, A.; Dolegowska, S.

    2011-01-01

    Trace element concentrations in plant bioindicators are often determined to assess the quality of the environment. Instrumental methods used for trace element determination require digestion of samples. There are different methods of sample preparation for trace element analysis, and the selection of the best method should be fitted for the purpose of a study. Our hypothesis is that the method of sample preparation is important for interpretation of the results. Here we compare the results of 36 element determinations performed by ICP-MS on ashed and on acid-digested (HNO3, H2O2) samples of two moss species (Hylocomium splendens and Pleurozium schreberi) collected in Alaska and in south-central Poland. We found that dry ashing of the moss samples prior to analysis resulted in considerably lower detection limits of all the elements examined. We also show that this sample preparation technique facilitated the determination of interregional and interspecies differences in the chemistry of trace elements. Compared to the Polish mosses, the Alaskan mosses displayed more positive correlations of the major rock-forming elements with ash content, reflecting those elements' geogenic origin. Of the two moss species, P. schreberi from both Alaska and Poland was also highlighted by a larger number of positive element pair correlations. The cluster analysis suggests that the more uniform element distribution pattern of the Polish mosses primarily reflects regional air pollution sources. Our study has shown that the method of sample preparation is an important factor in statistical interpretation of the results of trace element determinations. ?? 2010 Springer-Verlag.

  11. MALDI MS sample preparation by using paraffin wax film: systematic study and application for peptide analysis.

    PubMed

    Wang, Junhua; Chen, Ruibing; Ma, Mingming; Li, Lingjun

    2008-01-15

    Recently developed sample preparation techniques employing hydrophobic sample support have improved the detection sensitivity and mass spectral quality of matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). These methods concentrate the samples on target by minimizing the sample area via the solvent repellent effect of the target surface. In the current study, we employed the use of paraffin wax film (Parafilm M) for improved MALDI MS analysis of low-abundance peptide mixtures, including neuronal tissue releasate and protein tryptic digests. This thin film was found to strongly repel polar solvents including water, methanol, and acetonitrile, which enabled the application of a wide range of sample preparation protocols that involved the use of various organic solvents. A "nanoliter-volume deposition" technique employing a capillary column has been used to produce tiny ( approximately 400 microm) matrix spots of 2,5-dihydroxybenzoic acid on the film. By systematically optimizing the sample volume, solvent composition, and film treatment, the Parafilm M substrate in combination with the nanoliter-volume matrix deposition method allowed dilute sample to be concentrated on the film for MALDI MS analysis. Peptide mixtures with nanomolar concentrations have been detected by MALDI time-of-flight and MALDI Fourier transform ion cyclotron resonance mass spectrometers. Overall, the use of Parafilm M enabled improved sensitivity and spectral quality for the analysis of complex peptide mixtures.

  12. Effect of sample preparation on charged impurities in graphene substrates

    NASA Astrophysics Data System (ADS)

    Burson, K. M.; Dean, C. R.; Watanabe, K.; Taniguchi, T.; Hone, J.; Kim, P.; Cullen, W. G.; Fuhrer, M. S.

    2013-03-01

    The mobility of graphene as fabricated on SiO2 has been found to vary widely depending on sample preparation conditions. Additionally, graphene mobility on SiO2 appears to be limited to ~20,000 cm2/Vs, likely due to charged impurities in the substrate. Here we present a study of the effect of fabrication procedures on substrate charged impurity density (nimp) utilizing ultrahigh-vacuum Kelvin probe force microscopy. We conclude that even minimal SEM exposure, as from e-beam lithography, induces an increased impurity density, while heating reduces the number of charges for sample substrates which already exhibit a higher impurity density. We measure both SiO2 and h-BN and find that all nimp values observed for SiO2 are higher than those observed for h-BN; this is consistent with the observed improvement in mobility for graphene devices fabricated on h-BN over those fabricated on SiO2 substrates. This work was supported by the US ONR MURI program, and the University of Maryland NSF-MRSEC under Grant No. DMR 05-20471.

  13. Automated acoustic matrix deposition for MALDI sample preparation.

    PubMed

    Aerni, Hans-Rudolf; Cornett, Dale S; Caprioli, Richard M

    2006-02-01

    Novel high-throughput sample preparation strategies for MALDI imaging mass spectrometry (IMS) and profiling are presented. An acoustic reagent multispotter was developed to provide improved reproducibility for depositing matrix onto a sample surface, for example, such as a tissue section. The unique design of the acoustic droplet ejector and its optimization for depositing matrix solution are discussed. Since it does not contain a capillary or nozzle for fluid ejection, issues with clogging of these orifices are avoided. Automated matrix deposition provides better control of conditions affecting protein extraction and matrix crystallization with the ability to deposit matrix accurately onto small surface features. For tissue sections, matrix spots of 180-200 microm in diameter were obtained and a procedure is described for generating coordinate files readable by a mass spectrometer to permit automated profile acquisition. Mass spectral quality and reproducibility was found to be better than that obtained with manual pipet spotting. The instrument can also deposit matrix spots in a dense array pattern so that, after analysis in a mass spectrometer, two-dimensional ion images may be constructed. Example ion images from a mouse brain are presented.

  14. Miniaturized sample preparation method for determination of amphetamines in urine.

    PubMed

    Nishida, Manami; Namera, Akira; Yashiki, Mikio; Kimura, Kojiro

    2004-07-16

    A simple and miniaturized sample preparation method for determination of amphetamines in urine was developed using on-column derivatization and gas chromatography-mass spectrometry (GC-MS). Urine was directly applied to the extraction column that was pre-packed with Extrelut and sodium carbonate. Amphetamine (AP) and methamphetamine (MA) in urine were adsorbed on the surface of Extrelut. AP and MA were then converted to a free base and derivatized to N-propoxycarbonyl derivatives using propylchloroformate on the column. Pentadeuterated MA was used as an internal standard. The recoveries of AP and MA from urine were 100 and 102%, respectively. The calibration curves showed linearity in the range of 0.50-50 microg/mL for AP and MA in urine. When urine samples containing two different concentrations (0.50 and 5.0 microg/mL) of AP and MA were determined, the intra-day and inter-day coefficients of variation were 1.4-7.7%. This method was applied to 14 medico-legal cases of MA intoxication. The results were compared and a good agreement was obtained with a HPLC method.

  15. The Comparison of Hydrochloric Acid and Phosphoric Acid Treatments in the Preparation of Montmorillonite Catalysts for RNA Synthesis.

    PubMed

    Aldersley, Michael Frank; Joshi, Prakash C; Huang, Yixing

    2017-02-16

    The treatment of clay minerals with a preliminary acid wash and titration to pH 7 has proven to generate catalysts for the most interesting of oligomerization reactions in which activated RNA-nucleotides generate oligomers up to 40-mers. Significantly, not all clay minerals become catalytic following this treatment and none are catalytic in the absence of such treatment. The washing procedure has been modified and explored further using phosphoric acid and the outcomes are compared to those obtained when clay samples are prepared following a hydrochloric acid wash.

  16. The Comparison of Hydrochloric Acid and Phosphoric Acid Treatments in the Preparation of Montmorillonite Catalysts for RNA Synthesis

    NASA Astrophysics Data System (ADS)

    Aldersley, Michael Frank; Joshi, Prakash C.; Huang, Yixing

    2017-02-01

    The treatment of clay minerals with a preliminary acid wash and titration to pH 7 has proven to generate catalysts for the most interesting of oligomerization reactions in which activated RNA-nucleotides generate oligomers up to 40-mers. Significantly, not all clay minerals become catalytic following this treatment and none are catalytic in the absence of such treatment. The washing procedure has been modified and explored further using phosphoric acid and the outcomes are compared to those obtained when clay samples are prepared following a hydrochloric acid wash.

  17. Portable system for microbial sample preparation and oligonucleotide microarray analysis.

    SciTech Connect

    Bavykin, S. G.; Akowski, J. P.; Zakhariev, V. M.; Barsky, V. E.; Mirzabekov, A. D.; Perov, A. N.; Biochip Technology Center; Engelhardt Inst. of Molecular Biology

    2001-02-01

    We have developed a three-component system for microbial identification that consists of (i) a universal syringe-operated silica minicolumn for successive DNA and RNA isolation, fractionation, fragmentation, fluorescent labeling, and removal of excess free label and short oligonucleotides; (ii) microarrays of immobilized oligonucleotide probes for 16S rRNA identification; and (iii) a portable battery-powered device for imaging the hybridization of fluorescently labeled RNA fragments with the arrays. The minicolumn combines a guanidine thiocyanate method of nucleic acid isolation with a newly developed hydroxyl radical-based technique for DNA and RNA labeling and fragmentation. DNA and RNA can also be fractionated through differential binding of double- and single-stranded forms of nucleic acids to the silica. The procedure involves sequential washing of the column with different solutions. No vacuum filtration steps, phenol extraction, or centrifugation is required. After hybridization, the overall fluorescence pattern is captured as a digital image or as a Polaroid photo. This three-component system was used to discriminate Escherichia coli, Bacillus subtilis, Bacillus thuringiensis, and human HL60 cells. The procedure is rapid: beginning with whole cells, it takes approximately 25 min to obtain labeled DNA and RNA samples and an additional 25 min to hybridize and acquire the microarray image using a stationary image analysis system or the portable imager.

  18. Sample Preparation Report of the Fourth OPCW Confidence Building Exercise on Biomedical Sample Analysis

    SciTech Connect

    Udey, R. N.; Corzett, T. H.; Alcaraz, A.

    2014-07-03

    Following the successful completion of the 3rd biomedical confidence building exercise (February 2013 – March 2013), which included the analysis of plasma and urine samples spiked at low ppb levels as part of the exercise scenario, another confidence building exercise was targeted to be conducted in 2014. In this 4th exercise, it was desired to focus specifically on the analysis of plasma samples. The scenario was designed as an investigation of an alleged use of chemical weapons where plasma samples were collected, as plasma has been reported to contain CWA adducts which remain present in the human body for several weeks (Solano et al. 2008). In the 3rd exercise most participants used the fluoride regeneration method to analyze for the presence of nerve agents in plasma samples. For the 4th biomedical exercise it was decided to evaluate the analysis of human plasma samples for the presence/absence of the VX adducts and aged adducts to blood proteins (e.g., VX-butyrylcholinesterase (BuChE) and aged BuChE adducts using a pepsin digest technique to yield nonapeptides; or equivalent). As the aging of VX-BuChE adducts is relatively slow (t1/2 = 77 hr at 37 °C [Aurbek et al. 2009]), soman (GD), which ages much more quickly (t1/2 = 9 min at 37 °C [Masson et al. 2010]), was used to simulate an aged VX sample. Additional objectives of this exercise included having laboratories assess novel OP-adducted plasma sample preparation techniques and analytical instrumentation methodologies, as well as refining/designating the reporting formats for these new techniques.

  19. Acid digestion of geological and environmental samples using open-vessel focused microwave digestion.

    PubMed

    Taylor, Vivien F; Toms, Andrew; Longerich, Henry P

    2002-01-01

    The application of open vessel focused microwave acid digestion is described for the preparation of geological and environmental samples for analysis using inductively coupled plasma-mass spectrometry (ICP-MS). The method is compared to conventional closed-vessel high pressure methods which are limited in the use of HF to break down silicates. Open-vessel acid digestion more conveniently enables the use of HF to remove Si from geological and plant samples as volatile SiF4, as well as evaporation-to-dryness and sequential acid addition during the procedure. Rock reference materials (G-2 granite, MRG-1 gabbros, SY-2 syenite, JA-1 andesite, and JB-2 and SRM-688 basalts) and plant reference materials (BCR and IAEA lichens, peach leaves, apple leaves, Durham wheat flour, and pine needles) were digested with results comparable to conventional hotplate digestion. The microwave digestion method gave poor results for granitic samples containing refractory minerals, however fusion was the preferred method of preparation for these samples. Sample preparation time was reduced from several days, using conventional hotplate digestion method, to one hour per sample using our microwave method.

  20. The origin of amino acids in lunar regolith samples

    NASA Astrophysics Data System (ADS)

    Elsila, Jamie E.; Callahan, Michael P.; Dworkin, Jason P.; Glavin, Daniel P.; McLain, Hannah L.; Noble, Sarah K.; Gibson, Everett K.

    2016-01-01

    We analyzed the amino acid content of seven lunar regolith samples returned by the Apollo 16 and Apollo 17 missions and stored under NASA curation since collection using ultrahigh-performance liquid chromatography with fluorescence detection and time-of-flight mass spectrometry. Consistent with results from initial analyses shortly after collection in the 1970s, we observed amino acids at low concentrations in all of the curated samples, ranging from 0.2 parts-per-billion (ppb) to 42.7 ppb in hot-water extracts and 14.5-651.1 ppb in 6 M HCl acid-vapor-hydrolyzed, hot-water extracts. Amino acids identified in the Apollo soil extracts include glycine, D- and L-alanine, D- and L-aspartic acid, D- and L-glutamic acid, D- and L-serine, L-threonine, and L-valine, all of which had previously been detected in lunar samples, as well as several compounds not previously identified in lunar regoliths: α-aminoisobutyric acid (AIB), D- and L-β-amino-n-butyric acid (β-ABA), DL-α-amino-n-butyric acid, γ-amino-n-butyric acid, β-alanine, and ε-amino-n-caproic acid. We observed an excess of the L enantiomer in most of the detected proteinogenic amino acids, but racemic alanine and racemic β-ABA were present in some samples. We also examined seven samples from Apollo 15, 16, and 17 that had been previously allocated to a non-curation laboratory, as well as two samples of terrestrial dunite from studies of lunar module engine exhaust that had been stored in the same laboratory. The amino acid content of these samples suggested that contamination had occurred during non-curatorial storage. We measured the compound-specific carbon isotopic ratios of glycine, β-alanine, and L-alanine in Apollo regolith sample 70011 and found values of -21‰ to -33‰. These values are consistent with those seen in terrestrial biology and, together with the enantiomeric compositions of the proteinogenic amino acids, suggest that terrestrial biological contamination is a primary source of the

  1. A simple supported liquid hollow fiber membrane microextraction for sample preparation of trihalomethanes in water samples.

    PubMed

    Vora-adisak, Narongchai; Varanusupakul, Pakorn

    2006-07-21

    A simple and efficient liquid-phase microextraction (LPME) technique using a supported liquid hollow fiber membrane, in conjunction with gas chromatography-electron capture detector has been developed for extraction and determination of trihalomethanes (THMs) in water samples. THMs were extracted from water samples through an organic extracting solvent impregnated in the pores and filled inside the porous hollow fiber membrane. Our simple conditions were conducted at 35 degrees C with no stirring and no salt addition in order to minimize sample preparation steps. Parameters such as types of hollow fiber membranes, extracting solvents and extraction time were studied and optimized. The method exhibited enrichment factors ranged from 28- to 62-fold within 30 min extraction time. The linearity of the method ranged from 0.2 to 100 microg l(-1). The limits of detection were in the low microg l(-1) level, ranging between 0.01 and 0.2 microg l(-1). The recoveries of spiked THMs at 5 microg l(-1) in water were between 98 and 105% with relative standard deviations (RSDs) less than 4%. Furthermore, the method was applied for determination of THMs in drinking water and tap water samples was reported.

  2. [Preparation of ferulic acid, senkyunolide I and senkyunolide H from Ligusticum chuanxiong by preparative HPLC].

    PubMed

    Xiong, Yao-Kun; Liang, Shuang; Hong, Yan-Long; Yang, Xiu-Juan; Shen, Lan; Du, Yan; Feng, Yi

    2013-06-01

    Preparative HPLC was used to prepare ferulic acid, senkyunolide I and senkyunolide H from Ligusticum chuanxiong. The separation was conducted on a Shim-Pack Prep-ODS (20.0 mm x 250 mm, 5 microm) column with the mobile phase of methanol-0.2% glacial acetic acid (50:50)at the flow rate of 5 mL x min(-1). The detection wavelength was 278 nm, and the purity of each compound was detected by HPLC analysis. Spectral data analyses including UV, ESI-MS and NMR were used to identify their structures. This method is simple, fast, which is suitable for preparation of standard reference of ferulic acid, senkyunolide I and senkyunolide H from L. chuanxiong and can meet the requirement of new drug research and development.

  3. [Study on preparation process of artesunate polylactic acid microspheres].

    PubMed

    Pan, Xu-Wang; Wang, Wei; Fang, Hong-Ying; Wang, Fu-Gen; Cai, Zhao-Bin

    2013-12-01

    This study aims to investigate the preparation process and in vitro release behavior of artesunate polylactic acid microspheres, in order to prepare an artesunate polylactic acid (PLA) administration method suitable for hepatic arterial embolization. With PLA as the material and polyvinyl alcohol (PVA) as the emulsifier, O/W emulsion/solvent evaporation method was adopted to prepare artesunate polylactic acid microspheres, and optimize the preparation process. With drug loading capacity, encapsulation efficiency and particle size as indexes, a single factor analysis was made on PLA concentration, PVA concentration, drug loading ratio and stirring velocity. Through an orthogonal experiment, the optimal processing conditions were determined as follows: PLA concentration was 9. 0% , PVA concentration was 0. 9% , drug loading ratio was 1:2 and stirring velocity was 1 000 r x min(-1). According to the verification of the optimal process, microsphere size, drug loading and entrapment rate of artesunate polylactic acid microspheres were (101.7 +/- 0.37) microm, (30.8 +/- 0.84)%, (53.6 +/- 0.62)%, respectively. The results showed that the optimal process was so reasonable and stable that it could lay foundation for further studies.

  4. Preparation of .alpha., .beta.-unsaturated carboxylic acids and anhydrides

    DOEpatents

    Spivey, James Jerry; Gogate, Makarand Ratnakav; Zoeller, Joseph Robert; Tustin, Gerald Charles

    1998-01-01

    Disclosed is a process for the preparation of .alpha.,.beta.-unsaturated carboxylic acids and anhydrides thereof which comprises contacting formaldehyde or a source of formaldehyde with a carboxylic anhydride in the presence of a catalyst comprising mixed oxides of vanadium, phosphorus and, optionally, a third component selected from titanium, aluminum or, preferably silicon.

  5. Preparation of {alpha}, {beta}-unsaturated carboxylic acids and anhydrides

    DOEpatents

    Spivey, J.J.; Gogate, M.R.; Zoeller, J.R.; Tustin, G.C.

    1998-01-20

    Disclosed is a process for the preparation of {alpha},{beta}-unsaturated carboxylic acids and anhydrides thereof which comprises contacting formaldehyde or a source of formaldehyde with a carboxylic anhydride in the presence of a catalyst comprising mixed oxides of vanadium, phosphorus and, optionally, a third component selected from titanium, aluminum or, preferably silicon.

  6. Method for preparing 6-.beta.-halopenicillanic acids

    DOEpatents

    Hansen, Erik I.; Kran-Nielsen, Mogens P.; Von Daehne, Welf

    1989-01-01

    The present invention relates to a new and improved method for the preparation of a compound of the formula I ##STR1## in which R stands for halogen, giving rise to high yields of substantially pure 6.beta.-halopenicillanic acids, obtained in one step.

  7. Sample to answer: a fully integrated nucleic acid identification system for bacteria monitoring

    NASA Astrophysics Data System (ADS)

    Kim, Jungkyu; Elsnab, John; Johnson, Michael; Gale, Bruce K.

    2010-02-01

    A fully integrated microfluidic system was developed and incorporates an EC-MWCNT (electrochemical multiwalled carbon nanotube) sensor for the detection of bacteria. Sample metering, reagent metering and delivery was implemented with microvalves and pumps embedded inside the microfluidic system. The nucleic acid extraction was performed using microchannels controlled using automated platforms and a disposable microfluidic silica cartridge. The target samples were flowed and hybridized with probe ssDNA (single strand DNA) across the MWCNT-EC sensor (built on a silicon chip), which was embedded in a microfluidic cell. The 9-pad sensor was scanned before and after hybridization to measure the quantity of RNA (Ribonucleic acid) bound to the array surface. A rapid and accurate sample-in answer-out nucleic acid system was realized with automated volume metering, microfluidic sample preparation, and integrated nano-biosensors.

  8. [Preparation and spectroscopic properties of terbium polypropenic acid film].

    PubMed

    Wang, Xi-gui; Wu, Hong-ying; Weng, Shi-fu; Wu, Jin-guang

    2002-12-01

    The bonding-type rare earth polymers-polypropenic-acid terbium film was prepared through the bonding way. Three-dimension fluorescence spectra showed that the optimum excitation wavelength was 306 nm, the strongest emission wavelength was 544 nm. The terbium polypropenic-acid film showed the characteristic Tb3+ emission when excited at 306 nm due to 5D4-7FJ transition (J = 6, 5, 4 and 3). The emission maximum at 544 nm was ascribed to 5D4-7F5 transition of Tb3+ and presented strong green emission. The luminescent properties of Tb3+ were not affected by polymerism of propenic-acid and the transparency of polypropenic-acid in visible light region was not affected by the dopping Tb3+. The fluorescence properties and spectroscopic properties of the terbium polypropenic-acid were investigated by excitation spectrum, emission spectrum, IR, far-IR, and Raman spectrum.

  9. Preparation and evaluations in vitro of oxaliplatin polylactic acid nanoparticles.

    PubMed

    Cui, Zhaoyuan; Sun, Yong; Liu, Xiaohong; Ju, Fang; Chen, Qian; Gao, Wen; Wei, Haitian

    2013-08-01

    The oxaliplatin nanoparticles were prepared with polylactic acid matrix, orthogonal test was applied to optimize the prescriptions, and the qualities of oxaliplatin nanoparticles were characterized by the shape, particle size, encapsulation efficiency (EE), and drug loading (DL). Oxaliplatin nanoparticle was prepared by solution replacement method. The formation of 0.25% Tween80, DMF-water 1:8 (v/v), oxaliplatin-polylactic acid 1:5 (w/w), and 20 mg/ml polylactic acid showed the suitable EE (17.4 ± 0.47%), DL (3.52 ± 0.07%). We observed the shape of oxaliplatin nanoparticles through SEM. The average size of the particles was 120.5 ± 8.7 nm, which was detected by N5 submicron particle size analyzer.

  10. Analysis of Carbohydrate and Fatty Acid Marker Abundance in Ricin Toxin Preparations for Forensic Information

    SciTech Connect

    Colburn, Heather A.; Wunschel, David S.; Kreuzer-Martin, Helen W.; Moran, James J.; Antolick, Kathryn C.; Melville, Angela M.

    2010-07-15

    One challenge in the forensic analysis of ricin samples is determining the method and extent of sample preparation. Ricin purification from the source castor seeds is essentially a protein purification through removal of the non-protein fractions of the seed. Two major, non-protein constituents in the seed are the castor oil and carbohydrates. Ricinoleic acid is a relatively unique fatty acid in nature and is the most abundant component of castor oil, which comprises roughly half the seed weight. The carbohydrate component comprises roughly half of the remaining “mash” left after oil and hull removal. We used derivatization of carbohydrate and fatty acid markers followed by identification and quantification using gas chromatography/mass spectrometry (GC/MS) to assess compositional changes in ricin samples purified by different methods. The loss of ricinoleic acid indicated steps for oil removal had occurred. Changes to the carbohydrate content of the sample were also observed following protein precipitation. The differential loss of arabinose relative to mannose indicated removal of the major carbohydrate fraction of the seed and enrichment of the protein content. Taken together, these changes in fatty acid and carbohydrate abundance are indicative of the preparation method used for each sample.

  11. Preparation, characterization and catalytic properties of MCM-48 supported tungstophosphoric acid mesoporous materials for green synthesis of benzoic acid

    SciTech Connect

    Wu, Hai-Yan; Zhang, Xiao-Li; Chen, Xi; Chen, Ya; Zheng, Xiu-Cheng

    2014-03-15

    MCM-48 and tungstophosphoric acid (HPW) were prepared and applied for the synthesis of HPW/MCM-48 mesoporous materials. The characterization results showed that HPW/MCM-48 obtained retained the typical mesopore structure of MCM-48, and the textural parameters decreased with the increase loading of HPW. The catalytic oxidation results of benzyl alcohol and benzaldehyde with 30% H{sub 2}O{sub 2} indicated that HPW/MCM-48 was an efficient catalyst for the green synthesis of benzoic acid. Furthermore, 35 wt% HPW/MCM-48 sample showed the highest activity under the reaction conditions. Highlights: • 5–45 wt% HPW/MCM-48 mesoporous catalysts were prepared and characterized. • Their catalytic activities for the green synthesis of benzoic acid were investigated. • HPW/MCM-48 was approved to be an efficient catalyst. • 5 wt% HPW/MCM-48 exhibited the highest catalytic activity.

  12. Thermal-driven attachment of gold nanoparticles prepared with ascorbic acid onto indium tin oxide surfaces

    NASA Astrophysics Data System (ADS)

    Aziz, Md. Abdul; Oyama, Munetaka

    2013-05-01

    Thermal-driven attachment of gold nanoparticles (AuNPs), of which size was less than 50 nm, onto the surfaces of indium tin oxide (ITO) is reported as a new phenomenon. This was permitted by preparing AuNPs via the reduction of hydrogen tetrachloroaurate (HAuCl4) with ascorbic acid (AA). While the AuNPs prepared via the AA reduction sparsely attached on the surface of ITO even at room temperature, a heat-up treatment at ca. 75 °C caused denser attachment of AuNPs on ITO surfaces. The attached density and the homogeneity after the thermal treatment were better than those of AuNP/ITO prepared using 3-aminopropyl-trimethoxysilane linker molecules. The denser attachment was observed similarly both by the immersion of ITO samples after the preparations of AuNPs by AA and by the in situ preparation of AuNPs with AA together with ITO samples. Thus, it is considered that the thermal-driven attachment of AuNPs would occur after the formation of AuNPs in the aqueous solutions, not via the growth of AuNPs on ITO surfaces. The preparation of AuNPs with AA would be a key for the thermal-driven attachment because the same attachments were not observed for AuNPs prepared with citrate ions or commercially available tannic acid-capped AuNPs.

  13. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING LIQUID FOOD SAMPLES FOR ANALYSIS OF POLAR ORGANIC POLLUTANTS (SOP-5.29)

    EPA Science Inventory

    This SOP describes the extraction and preparation of a liquid food sample for analysis of acidic persistent organic pollutants such as acid herbicides, pentachlorphenol, and 3,5,6-trichloro-2-phenol. It covers the extraction, concentration and derivatization of samples that are t...

  14. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING SOLID FOOD SAMPLES FOR ANALYSIS OF POLAR ORGANIC POLLUTANTS (SOP-5.28)

    EPA Science Inventory

    This SOP describes the extraction and preparation of a solid food sample for analysis of acidic persistent organic pollutants such as acid herbicides, pentachlorphenol, and 3,5,6-trichloro-2-phenol. It covers the extraction, concentration and derivatization of samples that are to...

  15. Total airborne mold particle sampling: evaluation of sample collection, preparation and counting procedures, and collection devices.

    PubMed

    Godish, Diana; Godish, Thad

    2008-02-01

    This study was conducted to evaluate (i) procedures used to collect, prepare, and count total airborne mold spore/particle concentrations, and (ii) the relative field performance of three commercially available total airborne mold spore/particle sampling devices. Differences between factory and laboratory airflow calibration values of axial fan-driven sampling instruments (used in the study) indicated a need for laboratory calibration using a mass flow meter to ensure that sample results were accurately calculated. An aniline blue-amended Calberla's solution adjusted to a pH of 4.2-4.4 provided good sample mounting/counting results using Dow Corning high vacuum grease, Dow Corning 280A adhesive, and Dow Corning 316 silicone release spray for samples collected using mini-Burkard and Allergenco samplers. Count variability among analysts was most pronounced in 5% counts of relatively low mold particle deposition density samples and trended downward with increased count percentage and particle deposition density. No significant differences were observed among means of 5, 10, and 20% counts and among analysts; a significant interaction effect was observed between analysts' counts and particle deposition densities. Significantly higher mini-Burkard and Air-O-Cell total mold spore/particle counts for 600x vs. 400x (1.9 and 2.3 x higher, respectively), 1000x vs. 600x (1.9 and 2.2 x higher, respectively) and 1000x vs. 400x (3.6 and 4.6 x higher, respectively) comparisons indicated that 1000x magnification counts best quantified total airborne mold spore/particles using light microscopy, and that lower magnification counts may result in unacceptable underreporting of airborne mold spore/particle concentrations. Modest but significantly higher (1.2x) total mold spore concentrations were observed with Allergenco vs. mini-Burkard samples collected in co-located, concurrently operated sampler studies; moderate but significantly higher mini-Burkard count values (1.4x) were

  16. Hybrid molecularly imprinted polymers synthesized with 3-aminopropyltriethoxysilane-methacrylic acid monomer for miniaturized solid-phase extraction: A new and economical sample preparation strategy for determination of acyclovir in urine.

    PubMed

    Yan, Hongyuan; Wang, Mingyu; Han, Yehong; Qiao, Fengxia; Row, Kyung Ho

    2014-06-13

    The miniaturized molecularly imprinted solid-phase extraction (mini-MISPE) coupled with high-performance liquid chromatography was proposed for the determination of acyclovir in urine. 1.5-mL tapered plastic centrifuge tube filled with hybrid molecularly imprinted polymers (HMIPs) was used as the cartridge of mini-MISPE, and the HMIPs synthesized with 3-aminopropyltriethoxy silane-methacrylic acid as monomer exhibited good recognition and selectivity for acyclovir. Under the optimized condition, good linear calibration was obtained in a range of 0.5-15μgmL(-1) with the correlation coefficient of 0.9994, and the recoveries at three spiked levels were 91.6-103.3% in urine with the relative standard deviation (RSD) of ≤3.5%. Excellent intra-day and inter-day repeatability were achieved with RSD of ≤2.6% and 4.0% in three different concentrations. This method combined the advantages of HMIPs and mini-MISPE, and it could become an alternative tool for analyzing the residues of acyclovir in complex urine matrices.

  17. Back-etch method for plan view transmission electron microscopy sample preparation of optically opaque films.

    PubMed

    Yao, Bo; Coffey, Kevin R

    2008-04-01

    Back-etch methods have been widely used to prepare plan view transmission electron microscopy (TEM) samples of thin films on membranes by removal of the Si substrate below the membrane by backside etching. The conventional means to determine when to stop the etch process is to observe the color of the light transmitted through the sample, which is sensitive to the remaining Si thickness. However, most metallic films thicker than 75 nm are opaque, and there is no detectable color change prior to film perforation. In this paper, a back-etch method based on the observation of an abrupt change of optical reflection contrast is introduced as a means to determine the etch endpoint to prepare TEM samples for these films. As the acid etchant removes the Si substrate material a rough interface is generated. This interface becomes a relatively smooth and featureless region when the etchant reaches the membrane (film/SiO2). This featureless region is caused by the mirror reflection of the film plane (film/SiO2 interface) through the optically transparent SiO2 layer. The lower etch rate of SiO2 (compared with Si) gives the operator enough time to stop the etching without perforating the film. A clear view of the morphology and control of Si roughness during etching are critical to this method, which are discussed in detail. The procedures of mounting wax removal and sample rinsing are also described in detail, as during these steps damage to the membrane may easily occur without appropriate consideration. As examples, the preparation of 100-nm-thick Fe-based amorphous alloy thin film and 160-nm-thick Cu-thin film samples for TEM imaging is described.

  18. Adsorption of bile acid by chitosan salts prepared with cinnamic acid and analogue compounds.

    PubMed

    Murata, Yoshifumi; Nagaki, Kumiko; Kofuji, Kyouko; Sanae, Fujiko; Kontani, Hitoshi; Kawashima, Susumu

    2006-01-01

    A chitosan (CS) powder treated with cinnamic acid and an analogue compound (CN) was prepared as CS-CN. Using it, bile acid adsorption by CS-CN and the release of CN were investigated in vitro. When CS-CN was soaked in a taurocholate solution, it released CN and simultaneously adsorbed the bile acid. For CS-CN prepared with cinnamic acid, the amount of CN released was 0.286 +/- 0.001 mmol/g CS-CN; the amount of taurocholate adsorbed was 0.284 +/- 0.003 mmol/g CS-CN. These two functions were recognized on alginate or pectin gel beads containing CS-CN. The amount of released CN was altered extensively by the species of CN used for gel-bead preparation. Results suggest that CS-CN is a candidate for complementary medicine to prevent lifestyle-related diseases.

  19. Determination of the presence of hyaluronic acid in preparations containing amino acids: the molecular weight characterization.

    PubMed

    Bellomaria, A; Nepravishta, R; Mazzanti, U; Marchetti, M; Piccioli, P; Paci, M

    2014-10-15

    Several pharmaceutical preparations contain hyaluronic acid in the presence of a large variety of low molecular weight charged molecules like amino acids. In these mixtures, it is particularly difficult to determine the concentration and the molecular weight of the hyaluronic acid fragments. In fact zwitterionic compounds in high concentration behave by masking the hyaluronic acid due to the electrostatic interactions between amino acids and hyaluronic acid. In such conditions the common colorimetric test of the hyaluronic acid determination appears ineffective and in the (1)H NMR spectra the peaks of the polymer disappear completely. By a simple separation procedure the presence of hyaluronic acid was revealed by the DMAB test and (1)H NMR while its average molecular weight in the final product was determined by DOSY NMR spectroscopy alone. The latter determination is very important due to the healthy effects of some sizes of this polymer's fragments.

  20. Final LDRD report : development of sample preparation methods for ChIPMA-based imaging mass spectrometry of tissue samples.

    SciTech Connect

    Maharrey, Sean P.; Highley, Aaron M.; Behrens, Richard, Jr.; Wiese-Smith, Deneille

    2007-12-01

    The objective of this short-term LDRD project was to acquire the tools needed to use our chemical imaging precision mass analyzer (ChIPMA) instrument to analyze tissue samples. This effort was an outgrowth of discussions with oncologists on the need to find the cellular origin of signals in mass spectra of serum samples, which provide biomarkers for ovarian cancer. The ultimate goal would be to collect chemical images of biopsy samples allowing the chemical images of diseased and nondiseased sections of a sample to be compared. The equipment needed to prepare tissue samples have been acquired and built. This equipment includes an cyro-ultramicrotome for preparing thin sections of samples and a coating unit. The coating unit uses an electrospray system to deposit small droplets of a UV-photo absorbing compound on the surface of the tissue samples. Both units are operational. The tissue sample must be coated with the organic compound to enable matrix assisted laser desorption/ionization (MALDI) and matrix enhanced secondary ion mass spectrometry (ME-SIMS) measurements with the ChIPMA instrument Initial plans to test the sample preparation using human tissue samples required development of administrative procedures beyond the scope of this LDRD. Hence, it was decided to make two types of measurements: (1) Testing the spatial resolution of ME-SIMS by preparing a substrate coated with a mixture of an organic matrix and a bio standard and etching a defined pattern in the coating using a liquid metal ion beam, and (2) preparing and imaging C. elegans worms. Difficulties arose in sectioning the C. elegans for analysis and funds and time to overcome these difficulties were not available in this project. The facilities are now available for preparing biological samples for analysis with the ChIPMA instrument. Some further investment of time and resources in sample preparation should make this a useful tool for chemical imaging applications.

  1. Amphiphilic mediated sample preparation for micro-flow cytometry

    DOEpatents

    Clague, David S.; Wheeler, Elizabeth K.; Lee, Abraham P.

    2006-07-25

    A flow cytometer includes a flow cell for detecting the sample, an oil phase in the flow cell, a water phase in the flow cell, an oil-water interface between the oil phase and the water phase, a detector for detecting the sample at the oil-water interface, and a hydrophobic unit operatively connected to the sample. The hydrophobic unit is attached to the sample. The sample and the hydrophobic unit are placed in an oil and water combination. The sample is detected at the interface between the oil phase and the water phase.

  2. Amphiphilic mediated sample preparation for micro-flow cytometry

    DOEpatents

    Clague, David S.; Wheeler, Elizabeth K.; Lee, Abraham P.

    2009-03-17

    A flow cytometer includes a flow cell for detecting the sample, an oil phase in the flow cell, a water phase in the flow cell, an oil-water interface between the oil phase and the water phase, a detector for detecting the sample at the oil-water interface, and a hydrophobic unit operatively connected to the sample. The hydrophobic unit is attached to the sample. The sample and the hydrophobic unit are placed in an oil and water combination. The sample is detected at the interface between the oil phase and the water phase.

  3. Voltammetric Determination of Ferulic Acid Using Polypyrrole-Multiwalled Carbon Nanotubes Modified Electrode with Sample Application

    PubMed Central

    Abdel-Hamid, Refat; Newair, Emad F.

    2015-01-01

    A polypyrrole-multiwalled carbon nanotubes modified glassy carbon electrode-based sensor was devised for determination of ferulic acid (FA). The fabricated sensor was prepared electrochemically using cyclic voltammetry (CV) and characterized using CV and scanning electron microscope (SEM). The electrode shows an excellent electrochemical catalytic activity towards FA oxidation. Under optimal conditions, the anodic peak current correlates linearly to the FA concentration throughout the range of 3.32 × 10−6 to 2.59 × 10−5 M with a detection limit of 1.17 × 10−6 M (S/N = 3). The prepared sensor is highly selective towards ferulic acid without the interference of ascorbic acid. The sensor applicability was tested for total content determination of FA in a commercial popcorn sample and showed a robust functionality.

  4. Selecting Sample Preparation Workflows for Mass Spectrometry-Based Proteomic and Phosphoproteomic Analysis of Patient Samples with Acute Myeloid Leukemia

    PubMed Central

    Hernandez-Valladares, Maria; Aasebø, Elise; Selheim, Frode; Berven, Frode S.; Bruserud, Øystein

    2016-01-01

    Global mass spectrometry (MS)-based proteomic and phosphoproteomic studies of acute myeloid leukemia (AML) biomarkers represent a powerful strategy to identify and confirm proteins and their phosphorylated modifications that could be applied in diagnosis and prognosis, as a support for individual treatment regimens and selection of patients for bone marrow transplant. MS-based studies require optimal and reproducible workflows that allow a satisfactory coverage of the proteome and its modifications. Preparation of samples for global MS analysis is a crucial step and it usually requires method testing, tuning and optimization. Different proteomic workflows that have been used to prepare AML patient samples for global MS analysis usually include a standard protein in-solution digestion procedure with a urea-based lysis buffer. The enrichment of phosphopeptides from AML patient samples has previously been carried out either with immobilized metal affinity chromatography (IMAC) or metal oxide affinity chromatography (MOAC). We have recently tested several methods of sample preparation for MS analysis of the AML proteome and phosphoproteome and introduced filter-aided sample preparation (FASP) as a superior methodology for the sensitive and reproducible generation of peptides from patient samples. FASP-prepared peptides can be further fractionated or IMAC-enriched for proteome or phosphoproteome analyses. Herein, we will review both in-solution and FASP-based sample preparation workflows and encourage the use of the latter for the highest protein and phosphorylation coverage and reproducibility. PMID:28248234

  5. Quality analysis of salmon calcitonin in a polymeric bioadhesive pharmaceutical formulation: sample preparation optimization by DOE.

    PubMed

    D'Hondt, Matthias; Van Dorpe, Sylvia; Mehuys, Els; Deforce, Dieter; DeSpiegeleer, Bart

    2010-12-01

    A sensitive and selective HPLC method for the assay and degradation of salmon calcitonin, a 32-amino acid peptide drug, formulated at low concentrations (400 ppm m/m) in a bioadhesive nasal powder containing polymers, was developed and validated. The sample preparation step was optimized using Plackett-Burman and Onion experimental designs. The response functions evaluated were calcitonin recovery and analytical stability. The best results were obtained by treating the sample with 0.45% (v/v) trifluoroacetic acid at 60 degrees C for 40 min. These extraction conditions did not yield any observable degradation, while a maximum recovery for salmon calcitonin of 99.6% was obtained. The HPLC-UV/MS methods used a reversed-phase C(18) Vydac Everest column, with a gradient system based on aqueous acid and acetonitrile. UV detection, using trifluoroacetic acid in the mobile phase, was used for the assay of calcitonin and related degradants. Electrospray ionization (ESI) ion trap mass spectrometry, using formic acid in the mobile phase, was implemented for the confirmatory identification of degradation products. Validation results showed that the methodology was fit for the intended use, with accuracy of 97.4+/-4.3% for the assay and detection limits for degradants ranging between 0.5 and 2.4%. Pilot stability tests of the bioadhesive powder under different storage conditions showed a temperature-dependent decrease in salmon calcitonin assay value, with no equivalent increase in degradation products, explained by the chemical interaction between salmon calcitonin and the carbomer polymer.

  6. Aquatic hazard assessment of a commercial sample of naphthenic acids.

    PubMed

    Swigert, James P; Lee, Carol; Wong, Diana C L; White, Russell; Scarlett, Alan G; West, Charles E; Rowland, Steven J

    2015-04-01

    This paper presents chemical composition and aquatic toxicity characteristics of a commercial sample of naphthenic acids (NAs). Naphthenic acids are derived from the refining of petroleum middle distillates and can contribute to refinery effluent toxicity. NAs are also present in oil sands process-affected water (OSPW), but differences in the NAs compositions from these sources precludes using a common aquatic toxicity dataset to represent the aquatic hazards of NAs from both origins. Our chemical characterization of a commercial sample of NAs showed it to contain in order of abundance, 1-ring>2-ring>acyclic>3-ring acids (∼84%). Also present were monoaromatic acids (7%) and non-acids (9%, polyaromatic hydrocarbons and sulfur heterocyclic compounds). While the acyclic acids were only the third most abundant group, the five most abundant individual compounds were identified as C(10-14) n-acids (n-decanoic acid to n-tetradecanoic acid). Aquatic toxicity testing of fish (Pimephales promelas), invertebrate (Daphnia magna), algae (Pseudokirchneriella subcapitata), and bacteria (Vibrio fischeri) showed P. promelas to be the most sensitive species with 96-h LL50=9.0 mg L(-1) (LC50=5.6 mg L(-1)). Acute EL50 values for the other species ranged 24-46 mg L(-1) (EC50 values ranged 20-30 mg L(-1)). Biomimetic extraction via solid-phase-microextraction (BE-SPME) suggested a nonpolar narcosis mode of toxic action for D. magna, P. subcapitata, and V. fischeri. The BE analysis under-predicted fish toxicity, which indicates that a specific mode of action, besides narcosis, may be a factor for fishes.

  7. Preparation of Samples for Single-Worm Tracking

    PubMed Central

    Yemini, Eviatar; Kerr, Rex A.; Schafer, William R.

    2016-01-01

    Neurobiological research in genetically tractable organisms relies heavily on robust assays for behavioral phenotypes. The simple body plan of the nematode Caenorhabditis elegans makes it particularly amenable to the use of automated microscopy and image analysis to describe behavioral patterns quantitatively. This protocol first describes the preparation and use of media for growing and maintaining worms for tracking. The second part of the protocol describes how to prepare a single young adult worm for recording during video analysis. Although the protocol was developed for use in a single-worm tracker, it addresses factors important for the generation of reproducible, standardized images in all systems. PMID:22135667

  8. Hydrophobic starch nanocrystals preparations through crosslinking modification using citric acid.

    PubMed

    Zhou, Jiang; Tong, Jin; Su, Xingguang; Ren, Lili

    2016-10-01

    Biodegradable starch nanocrystals prepared by an acid treatment process were modified through crosslinking modification using citric acid as reactant by a dry reaction method. The occurrence of crosslinking modification was evaluated by Fourier transform infrared spectroscopy and swelling degree. X-ray diffraction, wettability tests and contact angle measurements were used to characterize the modified starch nanocrystals. It was found that the crosslinked starch nanocrystals displayed a higher affinity for low polar solvents such as dichloromethane. The surface of starch nanocrystals became more roughness after crosslinking modification with citric acid and the size decreased as revealed by scanning electron microscopy and dynamic light scattering results. XRD analysis showed that the crystalline structure of starch nanocrystals was basically not changed after the crosslinking modification with shorter heating time. The resulting hydrophobic starch nanocrystals are versatile precursors to the development of nanocomposites.

  9. A Method for Microalgae Proteomics Analysis Based on Modified Filter-Aided Sample Preparation.

    PubMed

    Li, Song; Cao, Xupeng; Wang, Yan; Zhu, Zhen; Zhang, Haowei; Xue, Song; Tian, Jing

    2017-04-11

    With the fast development of microalgal biofuel researches, the proteomics studies of microalgae increased quickly. A filter-aided sample preparation (FASP) method is widely used proteomics sample preparation method since 2009. Here, a method of microalgae proteomics analysis based on modified filter-aided sample preparation (mFASP) was described to meet the characteristics of microalgae cells and eliminate the error caused by over-alkylation. Using Chlamydomonas reinhardtii as the model, the prepared sample was tested by standard LC-MS/MS and compared with the previous reports. The results showed mFASP is suitable for most of occasions of microalgae proteomics studies.

  10. EVALUATION OF ARG-1 SAMPLES PREPARED BY CESIUM CARBONATE DISSOLUTION DURING THE ISOLOK SME ACCEPTABILITY TESTING

    SciTech Connect

    Edwards, T.; Hera, K.; Coleman, C.

    2011-12-05

    Evaluation of Defense Waste Processing Facility (DWPF) Chemical Process Cell (CPC) cycle time identified several opportunities to improve the CPC processing time. The Mechanical Systems & Custom Equipment Development (MS&CED) Section of the Savannah River National Laboratory (SRNL) recently completed the evaluation of one of these opportunities - the possibility of using an Isolok sampling valve as an alternative to the Hydragard valve for taking DWPF process samples at the Slurry Mix Evaporator (SME). The use of an Isolok for SME sampling has the potential to improve operability, reduce maintenance time, and decrease CPC cycle time. The SME acceptability testing for the Isolok was requested in Task Technical Request (TTR) HLW-DWPF-TTR-2010-0036 and was conducted as outlined in Task Technical and Quality Assurance Plan (TTQAP) SRNLRP-2011-00145. RW-0333P QA requirements applied to the task, and the results from the investigation were documented in SRNL-STI-2011-00693. Measurement of the chemical composition of study samples was a critical component of the SME acceptability testing of the Isolok. A sampling and analytical plan supported the investigation with the analytical plan directing that the study samples be prepared by a cesium carbonate (Cs{sub 2}CO{sub 3}) fusion dissolution method and analyzed by Inductively Coupled Plasma - Optical Emission Spectroscopy (ICP-OES). The use of the cesium carbonate preparation method for the Isolok testing provided an opportunity for an additional assessment of this dissolution method, which is being investigated as a potential replacement for the two methods (i.e., sodium peroxide fusion and mixed acid dissolution) that have been used at the DWPF for the analysis of SME samples. Earlier testing of the Cs{sub 2}CO{sub 3} method yielded promising results which led to a TTR from Savannah River Remediation, LLC (SRR) to SRNL for additional support and an associated TTQAP to direct the SRNL efforts. A technical report resulting

  11. Preview of the NASA NNWG NDE Sample Preparation Handbook

    NASA Technical Reports Server (NTRS)

    2010-01-01

    This viewgraph presents a step-by-step how-to fabrication documentation of every kind of sample that is fabricated for MSFC by UA Huntsville, including photos and illustrations. The tabulation of what kind of samples are being fabricated for what NDE method, detailed instructions/documentation of the inclusion/creation of defects, detailed specifications for materials, processes, and equipment, case histories and/or experiences with the different fabrication methods and defect inclusion techniques, discussion of pitfalls and difficulties associated with sample fabrication and defect inclusion techniques, and a discussion of why certain fabrication techniques are needed as related to the specific NDE methods are included in this presentation.

  12. Preparing monodisperse macromolecular samples for successful biological small-angle X-ray and neutron-scattering experiments.

    PubMed

    Jeffries, Cy M; Graewert, Melissa A; Blanchet, Clément E; Langley, David B; Whitten, Andrew E; Svergun, Dmitri I

    2016-11-01

    Small-angle X-ray scattering (SAXS) and small-angle neutron scattering (SANS) are techniques used to extract structural parameters and determine the overall structures and shapes of biological macromolecules, complexes and assemblies in solution. The scattering intensities measured from a sample contain contributions from all atoms within the illuminated sample volume, including the solvent and buffer components, as well as the macromolecules of interest. To obtain structural information, it is essential to prepare an exactly matched solvent blank so that background scattering contributions can be accurately subtracted from the sample scattering to obtain the net scattering from the macromolecules in the sample. In addition, sample heterogeneity caused by contaminants, aggregates, mismatched solvents, radiation damage or other factors can severely influence and complicate data analysis, so it is essential that the samples be pure and monodisperse for the duration of the experiment. This protocol outlines the basic physics of SAXS and SANS, and it reveals how the underlying conceptual principles of the techniques ultimately 'translate' into practical laboratory guidance for the production of samples of sufficiently high quality for scattering experiments. The procedure describes how to prepare and characterize protein and nucleic acid samples for both SAXS and SANS using gel electrophoresis, size-exclusion chromatography (SEC) and light scattering. Also included are procedures that are specific to X-rays (in-line SEC-SAXS) and neutrons, specifically preparing samples for contrast matching or variation experiments and deuterium labeling of proteins.

  13. Determination of malic acid and other C4 dicarboxylic acids in atmospheric aerosol samples.

    PubMed

    Röhrl, Andreas; Lammel, Gerhard

    2002-03-01

    An ion chromatographic method was developed which is able to separate five unsubstituted and hydroxy C4 dicarboxylic acids, succinic, malic, tartaric, maleic and fumaric acid, besides the other unsubstituted C2-C5 dicarboxylic acids, oxalic, malonic and glutaric acids, as well as inorganic ions in samples extracted from atmospheric particulate matter. By the application of this method it was found for both rural and urban sites and for various types of air masses that in the summer-time malic acid is the most prominent C4 diacid (64 ng m(-3) by average), exceeding succinic acid concentration (28 ng m(-3) by average) considerably. In winter-time considerably less, a factor of 4-15, C4 acids occurred and succinic acid was more concentrated than malic acid. Tartaric, fumaric and maleic acids were less concentrated (5.1, 5.0 and 4.5 ng m(-3) by average, respectively). Tartaric acid was observed for the first time in ambient air. The results indicate that in particular anthropogenic sources are important for the precursors of succinic, maleic and fumaric acids. Biogenic sources seem to influence the occurrence of malic acid significantly.

  14. Intelligent front-end sample preparation tool using acoustic streaming.

    SciTech Connect

    Cooley, Erika J.; McClain, Jaime L.; Murton, Jaclyn K.; Edwards, Thayne L.; Achyuthan, Komandoor E.; Branch, Darren W.; Clem, Paul Gilbert; Anderson, John Mueller; James, Conrad D.; Smith, Gennifer; Kotulski, Joseph Daniel

    2009-09-01

    We have successfully developed a nucleic acid extraction system based on a microacoustic lysis array coupled to an integrated nucleic acid extraction system all on a single cartridge. The microacoustic lysing array is based on 36{sup o} Y cut lithium niobate, which couples bulk acoustic waves (BAW) into the microchannels. The microchannels were fabricated using Mylar laminates and fused silica to form acoustic-fluidic interface cartridges. The transducer array consists of four active elements directed for cell lysis and one optional BAW element for mixing on the cartridge. The lysis system was modeled using one dimensional (1D) transmission line and two dimensional (2D) FEM models. For input powers required to lyse cells, the flow rate dictated the temperature change across the lysing region. From the computational models, a flow rate of 10 {micro}L/min produced a temperature rise of 23.2 C and only 6.7 C when flowing at 60 {micro}L/min. The measured temperature changes were 5 C less than the model. The computational models also permitted optimization of the acoustic coupling to the microchannel region and revealed the potential impact of thermal effects if not controlled. Using E. coli, we achieved a lysing efficacy of 49.9 {+-} 29.92 % based on a cell viability assay with a 757.2 % increase in ATP release within 20 seconds of acoustic exposure. A bench-top lysing system required 15-20 minutes operating up to 58 Watts to achieve the same level of cell lysis. We demonstrate that active mixing on the cartridge was critical to maximize binding and release of nucleic acid to the magnetic beads. Using a sol-gel silica bead matrix filled microchannel the extraction efficacy was 40%. The cartridge based magnetic bead system had an extraction efficiency of 19.2%. For an electric field based method that used Nafion films, a nucleic acid extraction efficiency of 66.3 % was achieved at 6 volts DC. For the flow rates we tested (10-50 {micro}L/min), the nucleic acid extraction

  15. 7 CFR 61.34 - Drawing and preparation of sample.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... forwarding to a licensed cottonseed chemist for analysis and grading. The duplicate shall be sealed and retained by the sampler until the original official sample shall have been analyzed by a licensed...

  16. 7 CFR 61.34 - Drawing and preparation of sample.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... forwarding to a licensed cottonseed chemist for analysis and grading. The duplicate shall be sealed and retained by the sampler until the original official sample shall have been analyzed by a licensed...

  17. 7 CFR 61.34 - Drawing and preparation of sample.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... forwarding to a licensed cottonseed chemist for analysis and grading. The duplicate shall be sealed and retained by the sampler until the original official sample shall have been analyzed by a licensed...

  18. 7 CFR 61.34 - Drawing and preparation of sample.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... forwarding to a licensed cottonseed chemist for analysis and grading. The duplicate shall be sealed and retained by the sampler until the original official sample shall have been analyzed by a licensed...

  19. 7 CFR 61.34 - Drawing and preparation of sample.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... forwarding to a licensed cottonseed chemist for analysis and grading. The duplicate shall be sealed and retained by the sampler until the original official sample shall have been analyzed by a licensed...

  20. Influence of sample preparation on the assay of isoflavones

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The complexity of sample matrices, coexistence of multiple forms of bioactive phytochemicals, and their interaction of with other cellular components pose a significant challenge for optimize extraction and accurate estimation of bioactive phytochemicals in foods and dietary supplements. This artic...

  1. Sample preparation of solid samples for metal determination by atomic spectroscopy - An overview and selected recent applications

    SciTech Connect

    Sneddon, J.; Hardaway, C.; Bobbadi, K.; Reddy, A.

    2006-07-01

    Classical methods involving dry dissolution, wet decomposition, and microwave methods for digestion/dissolution of solid samples for metal determination by various atomic spectroscopic techniques are discussed. Recent applications of solid sample preparation are presented including soils, sediments, food, cosmetics, oils, and coal.

  2. Atmospheric pressure microwave sample preparation procedure for the combined analysis of total phosphorus and kjeldahl nitrogen.

    PubMed

    Collins, L W; Chalk, S J; Kingston, H M

    1996-08-01

    An atmospheric pressure microwave digestion method has been developed for the combined analysis of total phosphorus and Kjeldahl nitrogen in complex matrices. In comparison to the digestion steps in EPA Methods 365.4 (total phosphorus) and 351.x (Kjeldahl nitrogen), this method requires less time, eliminates the need for a catalyst, and reduces the toxicity of the waste significantly. It employs a microwave-assisted digestion step, using refluxing borosilicate glass vessels at atmospheric pressure. Traditionally, this method has a time-consuming sample preparation step and generates toxic waste through the use of heavy metal catalysts. These advantages are gained by the combination of a high boiling point acid (sulfuric acid) and the application of focused microwave irradiation, which enhances the digestion process by direct energy coupling. NIST standard reference materials 1572 (citrus leaves), 1577a (bovine liver), and 1566 (oyster tissue) and tryptophan were analyzed to validate the method. Phosphorus concentrations were determined by the colorimetric ascorbic acid method outlined in EPA Method 365.3. Kjeldahl nitrogen concentrations were determined using EPA Method 351.1. The results of the analyses showed good precision and are in excellent agreement with the NIST published values for both elements.

  3. Extraction, separation, and intramolecular carbon isotope characterization of athabasca oil sands acids in environmental samples.

    PubMed

    Ahad, Jason M E; Pakdel, Hooshang; Savard, Martine M; Simard, Marie-Christine; Smirnoff, Anna

    2012-12-04

    Here we report a novel approach to extract, isolate, and characterize high molecular weight organic acids found in the Athabasca oil sands region using preparative capillary gas chromatography (PCGC) followed by thermal conversion/elemental analysis-isotope ratio mass spectrometry (TC/EA-IRMS). A number of different "naphthenic acids" surrogate standards were analyzed as were samples from the bitumen-rich unprocessed McMurray Formation, oil sands process water, groundwater from monitoring wells, and surface water from the Athabasca River. The intramolecular carbon isotope signature generated by online pyrolysis (δ(13)C(pyr)) showed little variation (±0.6‰) within any given sample across a large range of mass fractions separated by PCGC. Oil sand, tailings ponds, and deep McMurray Formation groundwater were significantly heavier (up to ∼9‰) compared to surface water and shallow groundwater samples, demonstrating the potential use of this technique in source apportionment studies.

  4. Effects of Sample Preparation on the Infrared Reflectance Spectra of Powders

    SciTech Connect

    Brauer, Carolyn S.; Johnson, Timothy J.; Myers, Tanya L.; Su, Yin-Fong; Blake, Thomas A.; Forland, Brenda M.

    2015-05-22

    While reflectance spectroscopy is a useful tool in identifying molecular compounds, laboratory measurement of solid (particularly powder) samples often is confounded by sample preparation methods. For example, both the packing density and surface roughness can have an effect on the quantitative reflectance spectra of powdered samples. Recent efforts in our group have focused on developing standard methods for measuring reflectance spectra that accounts for sample preparation, as well as other factors such as particle size and provenance. In this work, the effect of preparation method on sample reflectivity was investigated by measuring the directional-hemispherical spectra of samples that were hand-packed as well as pressed into pellets using an integrating sphere attached to a Fourier transform infrared spectrometer. The results show that the methods used to prepare the sample have a substantial effect on the measured reflectance spectra, as do other factors such as particle size.

  5. Preparation and characterization of uniform drug particles: dehydrocholic acid.

    PubMed

    Mohamed, Amr Ali; Matijević, Egon

    2012-02-15

    Two methods for the preparation of uniform dispersions of dehydrocholic acid of different morphologies are described. In the first case, the drug was dissolved in acetone and then re-precipitated by adding a non-solvent (either water or an aqueous stabilizer solution), which yielded rod-like particles. In the second procedure, spheres, consisting of small elongated subunits, were obtained by acidification of basic aqueous solutions of the drug. The resulting particles were characterized in terms of their structure and surface charge characteristics.

  6. Microsystem strategies for sample preparation in biological detection.

    SciTech Connect

    James, Conrad D.; Galambos, Paul C.; Bennett, Dawn Jonita; Manginell, Monica; Okandan, Murat; Acrivos, Andreas; Brozik, Susan Marie; Khusid, Boris

    2005-03-01

    The objective of this LDRD was to develop microdevice strategies for dealing with samples to be examined in biological detection systems. This includes three sub-components: namely, microdevice fabrication, sample delivery to the microdevice, and sample processing within the microdevice. The first component of this work focused on utilizing Sandia's surface micromachining technology to fabricate small volume (nanoliter) fluidic systems for processing small quantities of biological samples. The next component was to develop interfaces for the surface-micromachined silicon devices. We partnered with Micronics, a commercial company, to produce fluidic manifolds for sample delivery to our silicon devices. Pressure testing was completed to examine the strength of the bond between the pressure-sensitive adhesive layer and the silicon chip. We are also pursuing several other methods, both in house and external, to develop polymer-based fluidic manifolds for packaging silicon-based microfluidic devices. The second component, sample processing, is divided into two sub-tasks: cell collection and cell lysis. Cell collection was achieved using dielectrophoresis, which employs AC fields to collect cells at energized microelectrodes, while rejecting non-cellular particles. Both live and dead Staph. aureus bacteria have been collected using RF frequency dielectrophoresis. Bacteria have been separated from polystyrene microspheres using frequency-shifting dielectrophoresis. Computational modeling was performed to optimize device separation performance, and to predict particle response to the dielectrophoretic traps. Cell lysis is continuing to be pursued using microactuators to mechanically disrupt cell membranes. Novel thermal actuators, which can generate larger forces than previously tested electrostatic actuators, have been incorporated with and tested with cell lysis devices. Significant cell membrane distortion has been observed, but more experiments need to be conducted to

  7. Hydroxyapatite-phosphonoformic acid hybrid compounds prepared by hydrothermal method

    NASA Astrophysics Data System (ADS)

    Turki, Thouraya; Othmani, Masseoud; Bantignies, Jean-Louis; Bouzouita, Khaled

    2014-01-01

    Hydroxyapatites were prepared in the presence of different amounts of phosphonoformic acid (PFA) via the hydrothermal method. The obtained powders were characterized through chemical analysis, XRD, IR, 31P MAS-NMR, TEM, and TG-TDA. The XRD showed that the PFA did not affect the apatite composition. Indeed, only a reduction of the crystallite size was noted. After grafting of PFA, the IR spectroscopy revealed the appearance of new bands belonging to HPO42- and carboxylate groups of the apatite and organic moiety, respectively. Moreover, the 31P MAS-NMR spectra exhibited a peak with a low intensity assigned to the terminal phosphonate group of the organic moiety in addition to that of the apatite. Based on these results, a reaction mechanism involving the surface hydroxyl groups (tbnd Casbnd OH) of the apatite and the carboxyl group of the acid was proposed.

  8. A high acid mesoporous USY zeolite prepared by alumination

    NASA Astrophysics Data System (ADS)

    Ma, Jinghong; Kang, Yuhong; Ma, Ning; Hao, Wenming; Wang, Yan; Li, Ruifeng

    2013-01-01

    A high-acidity HUSY zeolite with mesoporous structure was prepared by alumination with a dilute aqueous NaAlO2 solution and characterized by XRD, N2 adsorption, IR framework vibration and 29Si MAS NMR methods. The results indicated the extra-framework aluminum was reinserted into the tetrahedral framework through isomorphic substitution of framework Si (0Al) sites by Al ions, whereas the crystal and micropore structure were unaltered. FTIR spectra of hydroxyl vibrations and pyridine adsorbed on realuminated zeolites showed that the number of Brønsted acid sites and strong Lewis acid sites increased whereas weak Lewis acid sites decreased twice. The mesoporous structure composed of inter-and intra-crystalline pores in the aluminated HUSY increased the external surface area of the zeolite, improving accessibility of molecules to the active sites and enhancing its catalytic ability. The realuminated HUSY zeolite supported with Ru catalyst exhibited a higher catalytic activity for benzene hydrogenation than the parent HUSY zeolite; the reaction rate in comparison to the mesozeolite increased by 5.5 times.

  9. Rapid sample preparation for determination of iron in tissue by closed-vessel digestion and microwave energy.

    PubMed

    Van Wyck, D B; Schifman, R B; Stivelman, J C; Ruiz, J; Martin, D

    1988-06-01

    We developed a rapid acid-digestion method for preparing tissue samples for iron determination. Specimens were digested in nitric acid and hydrogen peroxide under high temperature and pressure in closed Teflon vessels, with microwave energy. Analysis for iron in 25- to 250-mg portions of digested bovine liver powder (National Bureau of Standards Certified Reference Material no. 1577a) showed excellent linearity ([predicted] = 1.007[actual] - 0.166 micrograms per sample) and analytical recovery (98%). Precision (CV) was 5.4% when iron content was 10 micrograms per sample. Assaying split samples of mouse tissues, we found a close correlation between iron concentrations obtained with closed vs open vessels ([closed] = 0.878[open] + 68 micrograms/g, r = 0.994, range 400-4600 micrograms/g dry weight). In contrast to time-consuming conventional procedures for tissue dissolution, closed-vessel digestion with microwave energy dramatically shortens time for tissue preparation, minimizes use of caustic acid, reduces risk of sample loss or contamination, and yields accurate and reproducible results.

  10. Increased amplification success from forensic samples with locked nucleic acids.

    PubMed

    Ballantyne, Kaye N; van Oorschot, Roland A H; Mitchell, R John

    2011-08-01

    Inadequate sample quantities and qualities can commonly result in poor DNA amplification success rates for forensic case samples. In some instances, modifying the PCR protocol or components may assist profiling by overcoming inhibition, or reducing the threshold required for successful amplification and detection. Incorporation of locked nucleic acids (LNAs) into PCR primers has previously been shown to increase amplification success for a range of non-forensic sample types and applications. To investigate their use in a forensic context, the PCR primers for four commonly used STR loci have been redesigned to include LNA bases. The modified LNA primers provided significantly increased amplification success when compared to standard DNA primers, with both high-quality buccal samples and simulated forensic casework samples. Peak heights increased by as much as 5.75× for the singleplex amplifications. When incorporated into multiplexes, the LNA primers continued to outperform standard DNA primers, with increased ease of optimisation, and increased amplification success. The use of LNAs in PCR primers can greatly assist the profiling of a range of samples, and increase success rates from challenging forensic samples.

  11. Sample preparation and assay refinements for pathogen detection platforms

    NASA Astrophysics Data System (ADS)

    Lim, Daniel V.; Kearns, Elizabeth A.; Leskinen, Stephaney D.; Magaña, Sonia; Stroot, Joyce M.; Hunter, Dawn M.; Schlemmer, Sarah M.

    2009-02-01

    Food-borne and waterborne microbial pathogens are a potential problem in biowarfare and public health. Such pathogens can affect the health, combat readiness, and effectiveness of the warfighter in a battlefield environment and present potential threats to the civilian population through intentional or natural contamination of food and water. Conventional procedures to detect and identify microbial pathogens in food, water, and other materials can take days to perform and may provide inconclusive information. Research at the University of South Florida's Advanced Biosensors Laboratory (ABL) focuses on development of sample processing procedures and biosensor-based assays for rapid detection of biothreat agents. Rapid processing methods, including use of an automated concentrator of microorganisms in water, have been developed for complex matrix samples including ground beef, apple juice, produce, potable water and recreational water, enabling such samples to be directly tested by biosensor assays for target analytes. Bacillus atrophaeus spores and other bacteria can be concentrated from potable and recreational water at low levels with a dead-end hollow-fiber ultrafiltration concentration system. Target bacteria recovered by these processing procedures can be identified by evanescent wave, fiber optic biosensors or other detection platforms. Fiber optic biosensor assays have been improved to include subsequent PCR analysis and viability determination of captured target bacteria using broth enrichment and/or ATP luminescence.

  12. Sample preparation in determination of lead in garden vegetables by flame atomic absorption spectrophotometry.

    PubMed

    Preer, J R; Stephens, B R; Bland, C W

    1982-07-01

    Dry and wet ashing methods have been used in the analysis of garden vegetables for Pb. The reliability of wet ashing has been verified by the method of standard additions. Comparison of dry and wet ashing showed good agreement for a variety of garden vegetables. Sample size was more strictly limited for the wet-ashed samples, which led to lower sensitivity. Vegetable samples are commonly analyzed for a number of trace elements, which introduces additional constraints on sample preparation, notably because of Cd loss on dry ashing. Pretreatment with HNO3/H2SO4 ash aid eliminated Cd loss. Reliability of dry ashing with pretreatment was shown with NBS SRM Orchard Leaves, Pine Needles, Spinach, and Tomato Leaves. The analysis was insensitive to ashing temperature in the range 480-625 degrees C. A practical detection limit for the method is about 2 ppm Pb, dry weight basis (DWB). Care must be exercised to avoid contamination of the sample with lead at this level by improper handling. Segregation and acid washing of glassware and protection of the sample from contact with any object not demonstrably clean was necessary. No evidence was found of Pb contamination at this level from tap water washing of fresh vegetables, forced-air oven drying, or grinding with mortar and pestle. No special clean room facilities or laboratory air purification measures were used. Sensitivity was increased 3-fold by extraction with dithizone in CHCl3 followed by back-extraction into dilute HCl. Detection limits were not improved, however, because of variation in the extraction results. The instrumental method for assessing effective correction for back-ground absorbance showed adequate compensation, although comparison of direct and extractive determinations showed a small but significant difference between the methods of about 1 ppm Pb (DWB).

  13. Oxidation-resistant acidic resins prepared by partial carbonization as cocatalysts in synthesis of adipic acid.

    PubMed

    Wei, Huijuan; Li, Hongbian; Liu, Yangqing; Jin, Peng; Wang, Xiangyu; Li, Baojun

    2012-08-01

    The oxidation-resistant acidic resins are of great importance for the catalytic oxidation systems. In this paper, the oxidatively stable acidic resins are obtained from the cation ion exchange resins (CIERs) through the thermal treatment in N(2) atmosphere. The structure and properties of the thermally treated CIERs were characterized by chemical analysis, Fourier transform infrared (FT-IR) spectra, acid capacity measurement and scanning electron microscope (SEM). The thermally treated CIERs possess high acid capacity up to 4.09 mmol g(-1). A partial carbonization is observed in the thermal treatment process of CIERs, but the morphology of resin spheres maintains well. The as-prepared CIERs are used as solid acids to assist the hydrogen peroxide oxidation of cyclohexene to adipic acid (ADA) with tungstic acid as the catalyst precursor. The improved yields of ADA in the recycling reaction are obtained in the presence of acidic CIERs. Meanwhile, the unproductive decomposition of H(2)O(2) is effectively suppressed. The high yields of ADA (about 81%) are kept by the thermally treated CIERs even after the fifth cycle. The thermally treated CIERs exhibit excellent acid-catalytic performance and possess remarkable oxidation-resistant capability.

  14. Optimisation of a sample preparation method for the determination of fumonisin B(1) in rice.

    PubMed

    Petrarca, Mateus Henrique; Rodrigues, Maria Isabel; Rossi, Elizeu Antonio; de Sylos, Célia Maria

    2014-09-01

    A simple, rapid and cost-effective sample preparation method for the determination of fumonisin B1 in rice was optimised using a strategy of sequential experimental designs. Initially, a Plackett-Burman design was applied to select the statistically significant variables for the determination of fumonisin B1, and then, a central composite rotatable design was used to define the optimal conditions of these variables. The method involves extraction with a 50% acetonitrile aqueous solution and glacial acetic acid, liquid-liquid partitioning with addition of anhydrous sodium sulphate and sodium chloride, followed by dispersive SPE clean-up with diatomaceous earth. The final extract was analysed by HPLC-FLD after precolumn derivatisation with ortho-phthaldialdehyde. The optimised method was validated for selectivity, linearity, matrix effect, limits of detection and quantification, trueness, and precision, and then applied to commercial samples of polished rice. This is the first report of the occurrence of fumonisin B1 in commercial samples of polished rice from the Southeast region of Brazil.

  15. Sample Preparation for Mass Spectrometry Imaging of Plant Tissues: A Review

    PubMed Central

    Dong, Yonghui; Li, Bin; Malitsky, Sergey; Rogachev, Ilana; Aharoni, Asaph; Kaftan, Filip; Svatoš, Aleš; Franceschi, Pietro

    2016-01-01

    Mass spectrometry imaging (MSI) is a mass spectrometry based molecular ion imaging technique. It provides the means for ascertaining the spatial distribution of a large variety of analytes directly on tissue sample surfaces without any labeling or staining agents. These advantages make it an attractive molecular histology tool in medical, pharmaceutical, and biological research. Likewise, MSI has started gaining popularity in plant sciences; yet, information regarding sample preparation methods for plant tissues is still limited. Sample preparation is a crucial step that is directly associated with the quality and authenticity of the imaging results, it therefore demands in-depth studies based on the characteristics of plant samples. In this review, a sample preparation pipeline is discussed in detail and illustrated through selected practical examples. In particular, special concerns regarding sample preparation for plant imaging are critically evaluated. Finally, the applications of MSI techniques in plants are reviewed according to different classes of plant metabolites. PMID:26904042

  16. Microfluidic devices for DNA sequencing: sample preparation and electrophoretic analysis.

    PubMed

    Paegel, Brian M; Blazej, Robert G; Mathies, Richard A

    2003-02-01

    Modern DNA sequencing 'factories' have revolutionized biology by completing the human genome sequence, but in the race to completion we are left with inefficient, cumbersome, and costly macroscale processes and supporting facilities. During the same period, microfabricated DNA sequencing, sample processing and analysis devices have advanced rapidly toward the goal of a 'sequencing lab-on-a-chip'. Integrated microfluidic processing dramatically reduces analysis time and reagent consumption, and eliminates costly and unreliable macroscale robotics and laboratory apparatus. A microfabricated device for high-throughput DNA sequencing that couples clone isolation, template amplification, Sanger extension, purification, and electrophoretic analysis in a single microfluidic circuit is now attainable.

  17. Preparation, characterization and application in deep catalytic ODS of the mesoporous silica pillared clay incorporated with phosphotungstic acid.

    PubMed

    Li, Baoshan; Liu, Zhenxing; Liu, Jianjun; Zhou, Zhiyuan; Gao, Xiaohui; Pang, Xinmei; Sheng, Huiting

    2011-10-15

    Mesoporous silica pillared clay (SPC) materials with different contents of H(3)PW(12)O(40) (HPW) heteropoly acid were synthesized by introducing HPW into clay interlayer template in an acidic suspension using sol-gel method. Samples with similar HPW loadings were also prepared by impregnation method using SPC as the support. The results of the characterizations showed that HPW was dispersed more homogeneously in the encapsulated samples than in the impregnated samples. The encapsulated materials exhibited better catalytic performance than the impregnated samples in oxidative desulfurization of dibenzothiophene-containing model oil. The sulfur removal reached up to 98.6% for the model oil under the experiential conditions.

  18. Evaluation of Sampling and Sample Preparation Modifications for Soil Containing Metallic Residues

    DTIC Science & Technology

    2012-01-01

    Depth Interval Number of Increments per Sample Field Implementation Sample Tool Selection Collection of Soil Sample Sample Processing Air Drying...86 159 29 Palms (CA)4 IA  Artillery/ Bombing 100 6 100 x 100 RDX 9.4 3.9 4.8 5.6 2.1 38 Hill AFB (UT)5 TTA  Thermal treatment 100 3 100 x 100...representation of the overall experimental design. Soil samples were transported to CRREL and air -dried on alumi- num trays. Once air -dried, each

  19. Analysis of underivatized amino acids in geological samples using ion-pairing liquid chromatography and electrospray tandem mass spectrometry.

    PubMed

    Liu, De-Ling; Beegle, Luther W; Kanik, Isik

    2008-04-01

    The capability of detecting biomarkers, such as amino acids, in chemically complex field samples is essential to establishing the knowledge required to search for chemical signatures of life in future planetary explorations. However, due to the complexities of in situ investigations, it is important to establish a new analytical scheme that utilizes a minimal amount of sample preparation. This paper reports the feasibility of a novel and sensitive technique, which has been established to quantitate amino acids in terrestrial crust samples directly without derivatization using volatile ion-pairing liquid chromatography and tandem mass spectrometry equipped with an electrospray ionization source. Adequate separation of 20 underivatized amino acids was achieved on a C(18) capillary column within 26 min with nonafluoropentanoic acid (NFPA) as ion-pairing reagent. Each amino acid was identified from its retention time as well as from its characteristic parent-to-daughter ion transition. Using tandem mass spectrometry as a detection technique allows co-elution of some amino acids, as it is more specific than traditional spectrophotometric methods. In the present study, terrestrial samples collected from 3 different locations were analyzed for their water-extractable free amino acid contents, following the removal of metal and organic interferences via ion exchange procedures. This is the first time that amino acids in geological samples were directly determined quantitatively without complicated derivatization steps. Depending on the amino acid, the detection limits varied from 0.02 to 5.7 pmol with the use of a 1 microl sample injection loop.

  20. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING DERMAL WIPE AND SURFACE WIPE SAMPLES FOR ANALYSIS OF POLAR ORGANIC POLLUTANTS (SOP-5.27)

    EPA Science Inventory

    The method for extracting and preparing a dermal or surface wipe sample for analysis of acidic persistent organic pollutants is summarized in this standard operating procedure. It covers the extraction and concentration of samples that are to be analyzed by gas chromatography/mas...

  1. Current trends and challenges in sample preparation for metallic nanoparticles analysis in daily products and environmental samples: A review

    NASA Astrophysics Data System (ADS)

    De la Calle, Inmaculada; Menta, Mathieu; Séby, Fabienne

    2016-11-01

    Due to the increasing use of nanoparticles (NPs) in consumer products, it becomes necessary to develop different strategies for their detection, identification, characterization and quantification in a wide variety of samples. Since the analysis of NPs in consumer products and environmental samples is particularly troublesome, a detailed description of challenges and limitations is given here. This review mainly focuses on sample preparation procedures applied for the mostly used techniques for metallic and metal oxide NPs characterization in consumer products and most outstanding publications of biological and environmental samples (from 2006 to 2015). We summarize the procedures applied for total metal content, extraction/separation and/or preconcentration of NPs from the matrix, separation of metallic NPs from their ions or from larger particles and NPs' size fractionation. Sample preparation procedures specifically for microscopy are also described. Selected applications in cosmetics, food, other consumer products, biological tissues and environmental samples are presented. Advantages and inconveniences of those procedures are considered. Moreover, selected simplified schemes for NPs sample preparation, as well as usual techniques applied are included. Finally, promising directions for further investigations are discussed.

  2. A multi-step transmission electron microscopy sample preparation technique for cracked, heavily damaged, brittle materials.

    PubMed

    Weiss Brennan, Claire V; Walck, Scott D; Swab, Jeffrey J

    2014-12-01

    A new technique for the preparation of heavily cracked, heavily damaged, brittle materials for examination in a transmission electron microscope (TEM) is described in detail. In this study, cross-sectional TEM samples were prepared from indented silicon carbide (SiC) bulk ceramics, although this technique could also be applied to other brittle and/or multiphase materials. During TEM sample preparation, milling-induced damage must be minimized, since in studying deformation mechanisms, it would be difficult to distinguish deformation-induced cracking from cracking occurring due to the sample preparation. The samples were prepared using a site-specific, two-step ion milling sequence accompanied by epoxy vacuum infiltration into the cracks. This technique allows the heavily cracked, brittle ceramic material to stay intact during sample preparation and also helps preserve the true microstructure of the cracked area underneath the indent. Some preliminary TEM results are given and discussed in regards to deformation studies in ceramic materials. This sample preparation technique could be applied to other cracked and/or heavily damaged materials, including geological materials, archaeological materials, fatigued materials, and corrosion samples.

  3. Flavonoids and Phenolic Acids in Methanolic Extracts, Infusions and Tinctures from Commercial Samples of Lemon Balm.

    PubMed

    Arceusz, Agnieszka; Wesolowski, Marek; Ulewicz-Magulska, Beata

    2015-06-01

    The aim of this study was to quantify the levels of flavonoids (rutin, myricetin, quercetin, kaempferol) and phenolic acids (gallic, p-coumaric, rosmarinic, syringic, caffeic, chlorogenic, ellagic, ferulic) in lemon balm (Melissa officinalis L.) commonly used as a culinary, aromatic and medicinal herb. A rapid and reliable HPLC procedure was developed to determine the phenolic compounds in methanolic extracts, infusions and tinctures prepared from lemon balm. Except for myricetin and quercetin, as well as ellagic, gallic and rosmarinic acids, higher levels of the analytes under study were determined in the methanolic extracts (up to 22 mg/g of dry weight, DW), than in infusions (up to 5 mg/g DW). Tinctures were the poorest in flavonoids and phenolic acids (below 550 μg/g DW), except for ellagic and rosmarinic acids, which were quantified in tinctures at higher levels (mg/g DW). To sum up, the flavonoids were extracted more effectively in the infusions and tinctures than the phenolic acids. Statistically significant correlations were found between phenolic acids, possibly owing to similar biochemical pathways of the compounds. The hierarchical cluster and principal component analyses have also shown that the samples of lemon balm could be differentiated based on the levels of flavonoids and phenolic acids.

  4. Sulfuric acid intercalated graphite oxide for graphene preparation.

    PubMed

    Hong, Yanzhong; Wang, Zhiyong; Jin, Xianbo

    2013-12-06

    Graphene has shown enormous potential for innovation in various research fields. The current chemical approaches based on exfoliation of graphite via graphite oxide (GO) are potential for large-scale synthesis of graphene but suffer from high cost, great operation difficulties, and serious waste discharge. We report a facile preparation of graphene by rapid reduction and expansion exfoliation of sulfuric acid intercalated graphite oxide (SIGO) at temperature just above 100°C in ambient atmosphere, noting that SIGO is easily available as the immediate oxidation descendent of graphite in sulfuric acid. The oxygenic and hydric groups in SIGO are mainly removed through dehydration as catalyzed by the intercalated sulfuric acid (ISA). The resultant consists of mostly single layer graphene sheets with a mean diameter of 1.07 μm after dispersion in DMF. This SIGO process is reductant free, easy operation, low-energy, environmental friendly and generates graphene with low oxygen content, less defect and high conductivity. The provided synthesis route from graphite to graphene via SIGO is compact and readily scalable.

  5. Accurate analysis of taurine, anserine, carnosine and free amino acids in a cattle muscle biopsy sample.

    PubMed

    Imanari, Mai; Higuchi, Mikito; Shiba, Nobuya; Watanabe, Akira

    2010-06-01

    We have established an analysis method for some free amino acids (FAAs), as well as taurine (Tau), anserine (Ans) and carnosine (Car), in a fresh biopsy sample from cattle muscle. A series of model biopsy samples, corresponding to the mixtures of lean meat, fat and connective tissue, was prepared and showed high correlation coefficients between the compound concentration and the 3-methylhistidine (3-MeHis) content derived from hydrolysis of the biopsy sample (r = 0.74-0.95, P < 0.01). Interference from blood contamination could not be neglected, because the concentration of some FAAs in blood was comparable to that in muscle. However, it was possible to control the contamination of Tau, Ans, Car, glutamic acid, glutamine, asparatic acid and alanine to less than 5.0% when the blood contamination was controlled to less than 23%.These results suggest the necessity of measuring 3-MeHis as an index of lean meat and hemoglobin as an index of blood contamination when compounds in muscle biopsy samples are evaluated. We have carried out a series of these analyses using one biopsy sample and reveal differences in Tau, Ans, Car and some FAAs in beef muscle after different feeding regimes.

  6. Sample pretreatment and nucleic acid-based detection for fast diagnosis utilizing microfluidic systems.

    PubMed

    Wang, Jung-Hao; Wang, Chih-Hung; Lee, Gwo-Bin

    2012-06-01

    Recently, micro-electro-mechanical-systems (MEMS) technology and micromachining techniques have enabled miniaturization of biomedical devices and systems. Not only do these techniques facilitate the development of miniaturized instrumentation for biomedical analysis, but they also open a new era for integration of microdevices for performing accurate and sensitive diagnostic assays. A so-called "micro-total-analysis-system", which integrates sample pretreatment, transport, reaction, and detection on a small chip in an automatic format, can be realized by combining functional microfluidic components manufactured by specific MEMS technologies. Among the promising applications using microfluidic technologies, nucleic acid-based detection has shown considerable potential recently. For instance, micro-polymerase chain reaction chips for rapid DNA amplification have attracted considerable interest. In addition, microfluidic devices for rapid sample pretreatment prior to nucleic acid-based detection have also achieved significant progress in the recent years. In this review paper, microfluidic systems for sample preparation, nucleic acid amplification and detection for fast diagnosis will be reviewed. These microfluidic devices and systems have several advantages over their large-scale counterparts, including lower sample/reagent consumption, lower power consumption, compact size, faster analysis, and lower per unit cost. The development of these microfluidic devices and systems may provide a revolutionary platform technology for fast sample pretreatment and accurate, sensitive diagnosis.

  7. Advances in sample preparation in electromigration, chromatographic and mass spectrometric separation methods.

    PubMed

    Gilar, M; Bouvier, E S; Compton, B J

    2001-02-16

    The quality of sample preparation is a key factor in determining the success of analysis. While analysis of pharmaceutically important compounds in biological matrixes has driven forward the development of sample clean-up procedures in last 20 years, today's chemists face an additional challenge: sample preparation and analysis of complex biochemical samples for characterization of genotypic or phenotypic information contained in DNA and proteins. This review focuses on various sample pretreatment methods designed to meet the requirements for the analysis of biopolymers and small drugs in complex matrices. We discuss the advances in development of solid-phase extraction (SPE) sorbents, on-line SPE, membrane-based sample preparation, and sample clean-up of biopolymers prior to their analysis by mass spectrometry.

  8. Sample pretreatment for the capillary electrophoretic determination of organic acids in chromium(III) plating baths.

    PubMed

    Taraba, Lukáš; Křížek, Tomáš; Kubíčková, Anna; Coufal, Pavel

    2015-12-01

    This work deals with the development and optimization of the sample pretreatment and consequent electrophoretic analysis of two modern plating baths containing chromium(III) and either citric acid or oxalic acid. Some model mixtures containing known amounts of components of industrial baths have been prepared to simulate simplified bath matrices. Prior to analysis, a sample pretreatment consisting of the addition of some agents that could release acid from the stable chromium complex was tested. Determination of organic anions was accomplished by indirect UV detection. The best results were achieved by precipitation of chromium(III) hydroxide. The content of oxalate and citrate in real samples was calculated as 96.5% (SD 2.3%) and 97.3% (SD 0.8%), respectively, of the declared amount. Very good robustness of the method and satisfactory repeatability of migration time and peak area were obtained. This simple inexpensive method is suitable for routine determination of citric and oxalic acid in chromium(III)-based plating baths.

  9. SIMPLE, SENSITIVE AND SELECTIVE SPECTROPHOTOMETRIC ASSAY OF NAPROXEN IN PURE, PHARMACEUTICAL PREPARATION AND HUMAN SERUM SAMPLES.

    PubMed

    Alizadeh, Nina; Keyhanian, Fereshteh

    2015-01-01

    Two simple, rapid and sensitive spectrophotometric methods have been developed for the determination of naproxen in pure, pharmaceutical preparation and human serum samples. These methods are based on the formation of yellow ion-pair complexes between naproxen and two sulfophthalein acid dyes, namely bromocresol green (BCG method) and bromothymol blue (BTB method). The resulting complexes were measured at 424 nm (BCG method) and at 422 nm (BTB method). The effects of variables such as reagent concentration and reaction time were investigated to optimize the procedure. Beer's law was obeyed in the concentration range of 10-105 µg/mL and 5-85 µg/mL and the detection limits were found to be 0.347 and 0.31 µg/mL for BCG and BTB methods, respectively. The developed methods have been successfully applied for the determination of naproxen in bulk drugs, pharmaceutical formulations and human serum samples with good accuracy and precision. The results are comparable to those of reference methods, and hence are recommended for quality control and routine analysis.

  10. Solventless and solvent-minimized sample preparation techniques for determining currently used pesticides in water samples: a review.

    PubMed

    Tankiewicz, Maciej; Fenik, Jolanta; Biziuk, Marek

    2011-10-30

    The intensification of agriculture means that increasing amounts of toxic organic and inorganic compounds are entering the environment. The pesticides generally applied nowadays are regarded as some of the most dangerous contaminants of the environment. Their presence in the environment, especially in water, is hazardous because they cause human beings to become more susceptible to disease. For these reasons, it is essential to monitor pesticide residues in the environment with the aid of all accessible analytical methods. The analysis of samples for the presence of pesticides is problematic, because of the laborious and time-consuming operations involved in preparing samples for analysis, which themselves may be a source of additional contaminations and errors. To date, it has been standard practice to use large quantities of organic solvents in the sample preparation process; but as these solvents are themselves hazardous, solventless and solvent-minimized techniques are coming into use. This paper discusses the most commonly used over the last 15 years sample preparation techniques for monitoring organophosphorus and organonitrogen pesticides residue in water samples. Furthermore, a significant trend in sample preparation, in accordance with the principles of 'Green Chemistry' is the simplification, miniaturization and automation of analytical techniques. In view of this aspect, several novel techniques are being developed in order to reduce the analysis step, increase the sample throughput and to improve the quality and the sensitivity of analytical methods. The paper describes extraction techniques requiring the use of solvents - liquid-liquid extraction (LLE) and its modifications, membrane extraction techniques, hollow fibre-protected two-phase solvent microextraction, liquid phase microextraction based on the solidification of a floating organic drop (LPME-SFO), solid-phase extraction (SPE) and single-drop microextraction (SDME) - as well as solvent

  11. Novel techniques of preparing TEM samples for characterization of irradiation damage.

    PubMed

    Zhang, H K; Long, F; Yao, Z; Daymond, M R

    2013-12-01

    Focus ion beam preparation of transmission electron microscopy (TEM) samples has become increasingly popular due to the relative ease of extraction of TEM foils from specific locations within a larger sample. However the sputtering damage induced by Ga ion bombardment in focus ion beam means that traditional electropolishing may be a preferable method. First, we describe a special electropolishing method for the preparation of irregular TEM samples from ex-service nuclear reactor components, spring-shaped spacers. This method has also been used to prepare samples from a nonirradiated component for a TEM in situ heavy ion irradiation study. Because the specimen size is small (0.7 × 0.7 × 3 mm), a sandwich installation is adopted to obtain high quality polishing. Second, we describe some modifications to a conventional TEM cross-section sample preparation method that employs Ni electroplating. There are limitations to this method when preparing cross-section samples from either (1) metals which are difficult to activate for electroplating, or (2) a heavy ion irradiated foil with a very shallow damage layer close to the surface, which may be affected by the electroplating process. As a consequence, a novel technique for preparing cross-section samples was developed and is described.

  12. Electrodeposition as an alternate method for preparation of environmental samples for iodide by AMS

    SciTech Connect

    Adamic, M. L.; Lister, T. E.; Dufek, E. J.; Jenson, D. D.; Olson, J. E.; Vockenhuber, C.; Watrous, M. G.

    2015-03-25

    This paper presents an evaluation of an alternate method for preparing environmental samples for 129I analysis by accelerator mass spectrometry (AMS) at Idaho National Laboratory. The optimal sample preparation method is characterized by ease of preparation, capability of processing very small quantities of iodide, and ease of loading into a cathode. Electrodeposition of iodide on a silver wire was evaluated using these criteria. This study indicates that the electrochemically-formed silver iodide deposits produce ion currents similar to those from precipitated silver iodide for the same sample mass. Furthermore, precipitated silver iodide samples are usually mixed with niobium or silver powder prior to loading in a cathode. Using electrodeposition, the silver is already mixed with the sample and can simply be picked up with tweezers, placed in the sample die, and pressed into a cathode. The major advantage of this method is that the silver wire/electrodeposited silver iodide is much easier to load into a cathode.

  13. Microwave-assisted sample preparation of coal and coal fly ash for subsequent metal determination

    SciTech Connect

    Srogi, K.

    2007-01-15

    The aim of this paper is to review microwave-assisted digestion of coal and coal fly ash. A brief description of microwave heating principles is presented. Microwave-assisted digestion appears currently to be the most popular preparation technique, possibly due to the comparatively rapid sample preparation and the reduction of contamination, compared to the conventional hot-plate digestion methods.

  14. In-Situ Sample Preparation Development for Extraterrestrial Life Detection and Characterization

    NASA Astrophysics Data System (ADS)

    Craft, K. L.; Bradburne, C.; Tiffany, J.; Hagedon, M.; Hibbitts, C.; Vandegriff, J.; Horst, S.

    2017-02-01

    In-situ life detection instrumentation require robust sample preparation techniques that need further development in the coming years to enable the exciting life discoveries we seek in both familiar and unfamiliar planetary environments.

  15. Preparation of pure microbiological samples for pyrolysis gas-liquid chromatography studies

    NASA Technical Reports Server (NTRS)

    Oxborrow, G. S.; Fields, N. D.; Puleo, J. R.

    1976-01-01

    Bacterial samples were prepared for pyrolysis gas-liquid chromatography using cells grown on membrane filters. Pyrochromatograms were reproducible when cells harvested from the filters were pyrolyzed without being washed.

  16. Nanoparticle preparation of Mefenamic acid by electrospray drying

    SciTech Connect

    Zolkepali, Nurul Karimah Bakar, Noor Fitrah Abu Anuar, Nornizar; Naim, M. Nazli; Bakar, Mohd Rushdi Abu

    2014-02-24

    Nanoparticles preparation of Mefenamic acid (MA) by using an electrospray drying method was conducted in this study. Electrospray drying is a process that uses electrostatic force to disperse a conductive liquid stream into fine charged droplets through the coulomb fission of charges in the liquid and finally dry into fine particles. Electrospray drying modes operation usually in Taylor cone jet, and it was formed by controlling applied voltage and liquid flow rate. A conductive liquid (2.77–8.55μScm{sup −1}) which is MA solution was prepared by using acetone with concentration 0.041 and 0.055 M before pumping at a flow rate of 3–6ml/h. By applying the applied voltage at 1.3–1.5 kV, Taylor cone jet mode was formed prior to the electrospray. During electrospray drying process, solvent evaporation from the droplet was occurring that leads to coulomb disruption and may generate to nanoparticles. The dried nanoparticles were collected on a grounded substrate that was placed at varying distance from the electrospray. MA particle with size range of 100–400 nm were produced by electrospray drying process. Characterization of particles by using X-ray diffractometry (XRD) and differential scanning calorimetry (DSC) show that particles formed into polymorph I.

  17. THE PREPARATION AND PROPERTIES OF HIGHLY PURIFIED ASCORBIC ACID OXIDASE

    PubMed Central

    Powers, Wendell H.; Lewis, Stanley; Dawson, Charles R.

    1944-01-01

    1. A method is described for the preparation of a highly purified ascorbic acid oxidase containing 0.24 per cent copper. 2. Using comparable activity measurements, this oxidase is about one and a half times as active on a dry weight basis as the hitherto most highly purified preparation described by Lovett-Janison and Nelson. The latter contained 0.15 per cent copper. 3. The oxidase activity is proportional to the copper content and the proportionality factor is the same as that reported by Lovett-Janison and Nelson. 4. When dialyzed free of salt, the blue concentrated oxidase solutions precipitate a dark green-blue protein which carries the activity. This may be prevented by keeping the concentrated solutions about 0.1 M in Na2HPO4. 5. When highly diluted for activity measurements the oxidase rapidly loses activity (irreversibly) previous to the measurement, unless the dilution is made with a dilute inert protein (gelatin) solution. Therefore activity values obtained using such gelatin-stabilized dilute solutions of the oxidase run considerably higher than values obtained by the Lovett-Janison and Nelson technique. 6. The effect of pH and substrate concentration on the activity of the purified oxidase in the presence and absence of inert protein was studied. PMID:19873382

  18. Sample preparation with solid phase microextraction and exhaustive extraction approaches: Comparison for challenging cases.

    PubMed

    Boyacı, Ezel; Rodríguez-Lafuente, Ángel; Gorynski, Krzysztof; Mirnaghi, Fatemeh; Souza-Silva, Érica A; Hein, Dietmar; Pawliszyn, Janusz

    2015-05-11

    In chemical analysis, sample preparation is frequently considered the bottleneck of the entire analytical method. The success of the final method strongly depends on understanding the entire process of analysis of a particular type of analyte in a sample, namely: the physicochemical properties of the analytes (solubility, volatility, polarity etc.), the environmental conditions, and the matrix components of the sample. Various sample preparation strategies have been developed based on exhaustive or non-exhaustive extraction of analytes from matrices. Undoubtedly, amongst all sample preparation approaches, liquid extraction, including liquid-liquid (LLE) and solid phase extraction (SPE), are the most well-known, widely used, and commonly accepted methods by many international organizations and accredited laboratories. Both methods are well documented and there are many well defined procedures, which make them, at first sight, the methods of choice. However, many challenging tasks, such as complex matrix applications, on-site and in vivo applications, and determination of matrix-bound and free concentrations of analytes, are not easily attainable with these classical approaches for sample preparation. In the last two decades, the introduction of solid phase microextraction (SPME) has brought significant progress in the sample preparation area by facilitating on-site and in vivo applications, time weighted average (TWA) and instantaneous concentration determinations. Recently introduced matrix compatible coatings for SPME facilitate direct extraction from complex matrices and fill the gap in direct sampling from challenging matrices. Following introduction of SPME, numerous other microextraction approaches evolved to address limitations of the above mentioned techniques. There is not a single method that can be considered as a universal solution for sample preparation. This review aims to show the main advantages and limitations of the above mentioned sample

  19. Inert sampling and sample preparation--the influence of oxygen on heavy metal mobility in river sediments.

    PubMed

    Einax, J W; Nischwitz, V

    2001-11-01

    Two approaches have been used to investigate changes in the nature of metal binding in river sediments caused by atmospheric oxygen. Firstly, non-inert and inert sample preparation were applied, in combination with sequential extraction, to determine for which metals inert sample preparation is necessary for correct determination of metal mobility under environmental conditions. Secondly, the metal contents of sediments sampled before and after a river weir were fractionated by sequential extraction to study the effect of the oxygen impact at the weir on heavy metal mobility in the sediments. Different grain-size fractions from one sample were also extracted, to enable selection of the upper grain-size limit most suitable for answering this analytical question. The results showed the need for the inert sample preparation technique for Cd, Zn, Pb, Mn, and Fe, but not for Co, Ni, Cu, and Cr. No significant change of heavy metal mobility at the weir could be proved, although the mobilization behavior of some elements was different. The optimum upper grain-size limit was 63 microm.

  20. Sample preparation method for glass welding by ultrashort laser pulses yields higher seam strength

    SciTech Connect

    Cvecek, K.; Miyamoto, I.; Strauss, J.; Wolf, M.; Frick, T.; Schmidt, M.

    2011-05-01

    Glass welding by ultrashort laser pulses allows joining without the need of an absorber or a preheating and postheating process. However, cracks generated during the welding process substantially impair the joining strength of the welding seams. In this paper a sample preparation method is described that prevents the formation of cracks. The measured joining strength of samples prepared by this method is substantially higher than previously reported values.

  1. Proteomic Challenges: Sample Preparation Techniques for Microgram-Quantity Protein Analysis from Biological Samples

    PubMed Central

    Feist, Peter; Hummon, Amanda B.

    2015-01-01

    Proteins regulate many cellular functions and analyzing the presence and abundance of proteins in biological samples are central focuses in proteomics. The discovery and validation of biomarkers, pathways, and drug targets for various diseases can be accomplished using mass spectrometry-based proteomics. However, with mass-limited samples like tumor biopsies, it can be challenging to obtain sufficient amounts of proteins to generate high-quality mass spectrometric data. Techniques developed for macroscale quantities recover sufficient amounts of protein from milligram quantities of starting material, but sample losses become crippling with these techniques when only microgram amounts of material are available. To combat this challenge, proteomicists have developed micro-scale techniques that are compatible with decreased sample size (100 μg or lower) and still enable excellent proteome coverage. Extraction, contaminant removal, protein quantitation, and sample handling techniques for the microgram protein range are reviewed here, with an emphasis on liquid chromatography and bottom-up mass spectrometry-compatible techniques. Also, a range of biological specimens, including mammalian tissues and model cell culture systems, are discussed. PMID:25664860

  2. Two Dimensional Polyamides Prepared From Unsaturated Carboxylic Acids And Amines.

    DOEpatents

    McDonald, William F.; Huang, Zhi Heng; Wright, Stacy C.; Danzig, Morris; Taylor, Andrew C.

    2002-07-17

    A polyamide and a process for preparing the polyamide are disclosed. The process comprises reacting in a reaction mixture a monomer selected from unsaturated carboxylic acids, esters of unsaturated carboxylic acids, anhydrides of unsaturated carboxylic acids, and mixtures thereof, and a first amine to form an intermediate reaction product in the reaction mixture, wherein the first amine is selected from RR.sub.1 NH, RNH.sub.2, RR.sub.1 NH.sub.2.sup.+, RNH.sub.3.sup.+ and mixtures thereof, wherein R and R.sub.1 can be the same or different and each contain between about 1 and 50 carbon atoms and are optionally substituted with heteroatoms oxygen, nitrogen, sulfur, and phosphorus and combinations thereof, and reacting the intermediate reaction product and a second amine to form a polyamide, wherein the second amine is selected from R.sub.2 R.sub.3 NH, R.sub.2 NH.sub.2, R.sub.2 R.sub.3 NH.sub.2.sup.+, R.sub.2 NH.sub.3.sup.+ and mixtures thereof wherein R.sub.2 and R.sub.3 can be the same or different and each contain between about 1 and 50 carbon atoms and are optionally substituted with heteroatoms oxygen, nitrogen, sulfur, and phosphorus and combinations thereof, wherein multiple of the R, R.sub.1, R.sub.2, and R.sub.3 are in vertically aligned spaced relationship along a backbone formed by the polyamide. In one version of the invention, the monomer is selected from maleic anhydride, maleic acid esters, and mixtures thereof. In another version of the invention, the first amine is an alkylamine, such as tetradecylamine, and the second amine is a polyalkylene polyamine, such as pentaethylenehexamine. In yet another version of the invention, the first amine and the second amine are olefinic or acetylenic amines, such as the reaction products of an alkyldiamine and an acetylenic carboxylic acid. The first amine and the second amine may be the same or different depending on the desired polyamide polymer structure.

  3. Sample Preparation and Identification of Biological, Chemical and Mid-Spectrum Agents

    DTIC Science & Technology

    2005-10-01

    mL sterile PBS. Form a suspension by vigorous agitation using a vortex mixer for 10-15 seconds. Soil and Other Solid Environmental Samples. To 5...grams of sample add two equivalent volumes of sterile PBS. Form a suspension by vigorous agitation on vortex mixer for 10-15 seconds. Filter sample...Micro-Extraction Solid phase microextraction (SPME) is a relatively new analytical technique. It can combine sample preparation and GC or LC analysis in

  4. Electrophilic amination of amino acids with N-Boc-oxaziridines: efficient preparation of N-orthogonally diprotected hydrazino acids and piperazic acid derivatives.

    PubMed

    Hannachi, Jean-Christophe; Vidal, Joëlle; Mulatier, Jean-Christophe; Collet, André

    2004-04-02

    A general two-step preparation of enantiopure N(alpha),N(beta)-orthogonally diprotected alpha-hydrazino acids 1 is developed on a multigram scale. The key reaction is the efficient electrophilic amination of N-benzyl amino acids 6 with N-Boc-oxaziridine 7 and accommodates various functional groups encountered in side chains of amino acids. The cyclic 2,3,4,5-tetrahydro-3-pyridazine carboxylic acid (piperazic acid) derivatives 2 and 3 or the cyclic 3,4-dihydro-3-pyrazolecarboxylate 4 are conveniently prepared from glutamic acid or aspartic acid via orthogonally diprotected alpha-hydrazino acids 1m and 1n.

  5. Coagulant properties of Moringa oleifera protein preparations: application to humic acid removal.

    PubMed

    Santos, Andréa F S; Paiva, Patrícia M G; Teixeira, José A C; Brito, António G; Coelho, Luana C B B; Nogueira, Regina

    2012-01-01

    This work aimed to characterize the coagulant properties of protein preparations from Moringa oleifera seeds in the removal of humic acids from water. Three distinct preparations were assayed, namely extract (seeds homogenized with 0.15 M NaCl), fraction (extract precipitated with 60% w/v ammonium sulphate) and cMoL (protein purified with guar gel column chromatography). The extract showed the highest coagulant activity in a protein concentration between 1 mg/L and 180 mg/L at pH 7.0. The zeta potential of the extract (-10 mV to -15 mV) was less negative than that of the humic acid (-41 mV to -42 mV) in a pH range between 5.0 and 8.0; thus, the mechanism that might be involved in this coagulation activity is adsorption and neutralization of charges. Reduction of total organic carbon (TOC) and dissolved organic carbon (DOC) was observed in water samples containing 9 mg/L carbon as humic acid when treated with 1 mg/L of the extract. A decrease in colour and in the aromatic content of the treated water was also observed. These results suggested that the extract from M. oleifera seeds in a low concentration (1 mg/L) can be an interesting natural alternative for removing humic acid from water in developing countries. The extract dose determined in the present study does not impart odour or colour to the treated water.

  6. Chiral speciation of selenoamino acids in biological samples.

    PubMed

    Chen, Beibei; He, Man; Zhong, Cheng; Hu, Bin

    2014-10-10

    In this paper, the "state of the art" of chiral speciation of selenoamino acids (SeAAs) in biological samples is critically reviewed. The significance and the features of such studies are highlighted. A special focus lies on chiral speciation of SeAAs by hyphenation techniques in which a chiral separation method (such as gas chromatography (GC), high performance liquid chromatography (HPLC) and capillary electrophoresis (CE)) is on-line coupled with an elemental specific detector, especially inductively coupled plasma mass spectrometry (ICP-MS). The advances in the development and application of hyphenation techniques in chiral speciation of SeAAs in biological samples are summarized and a perspective for future developments including sophisticated and innovative applications is discussed. Overall, HPLC-ICP-MS is more applicable than GC/CE-ICP-MS for chiral speciation of SeAAs. In the future, more novel chiral HPLC methods with high enantio-resolution, low cost and robustness, and their more applications in real biological samples analysis are expected.

  7. Sample preparation for mass spectrometry imaging: small mistakes can lead to big consequences.

    PubMed

    Goodwin, Richard J A

    2012-08-30

    Mass spectrometry imaging (MSI) enables the direct analysis of molecules from the surface of a wide variety of samples, allowing the multiplex measurement of both abundance and distribution of small molecules, lipids, peptides and proteins. As the technology has been refined an increasing number of ionization methods and mass analyzers has been used that enable increased spatial and spectral resolution measurements to be made at an increased speed. Alongside the instrumentation improvements there has been optimization of sample preparation procedures that allow the highest quality data to be obtained, reproducibly, from an ever increasing diversity of samples. This review will consider the development and standardization of sample preparation methods applicable to MSI, describing the stages and procedures undertaken from the instance of sample collection, through storage, preparation and on through final processing prior to analysis. Recent technical advancements will be highlighted and areas where further experimentation and optimization may well be required will be described. All aspects of the sample preparation pipeline will be considered in detail, with examples from the literature used to emphasize why rigorous sample preparation for MSI is vital to achieve the most accurate, reproducible and validated MSI data possible.

  8. ANALYSIS OF RICIN TOXIN PREPARATIONS FOR CARBOHYDRATE AND FATTY ACID ABUNDANCE AND ISOTOPE RATIO INFORMATION

    SciTech Connect

    Wunschel, David S.; Kreuzer-Martin, Helen W.; Antolick, Kathryn C.; Colburn, Heather A.; Moran, James J.; Melville, Angela M.

    2009-12-01

    This report describes method development and preliminary evaluation for analyzing castor samples for signatures of purifying ricin. Ricin purification from the source castor seeds is essentially a problem of protein purification using common biochemical methods. Indications of protein purification will likely manifest themselves as removal of the non-protein fractions of the seed. Two major, non-protein, types of biochemical constituents in the seed are the castor oil and various carbohydrates. The oil comprises roughly half the seed weight while the carbohydrate component comprises roughly half of the remaining “mash” left after oil and hull removal. Different castor oil and carbohydrate components can serve as indicators of specific toxin processing steps. Ricinoleic acid is a relatively unique fatty acid in nature and is the most abundant component of castor oil. The loss of ricinoleic acid indicates a step to remove oil from the seeds. The relative amounts of carbohydrates and carbohydrate-like compounds, including arabinose, xylose, myo-inositol fucose, rhamnose, glucosamine and mannose detected in the sample can also indicate specific processing steps. For instance, the differential loss of arabinose relative to mannose and N-acetyl glucosamine indicates enrichment for the protein fraction of the seed using protein precipitation. The methods developed in this project center on fatty acid and carbohydrate extraction from castor samples followed by derivatization to permit analysis by gas chromatography-mass spectrometry (GC-MS). Method descriptions herein include: the source and preparation of castor materials used for method evaluation, the equipment and description of procedure required for chemical derivatization, and the instrument parameters used in the analysis. Two types of derivatization methods describe analysis of carbohydrates and one procedure for analysis of fatty acids. Two types of GC-MS analysis is included in the method development, one

  9. Solar-thermal complex sample processing for nucleic acid based diagnostics in limited resource settings

    PubMed Central

    Gumus, Abdurrahman; Ahsan, Syed; Dogan, Belgin; Jiang, Li; Snodgrass, Ryan; Gardner, Andrea; Lu, Zhengda; Simpson, Kenneth; Erickson, David

    2016-01-01

    The use of point-of-care (POC) devices in limited resource settings where access to commonly used infrastructure, such as water and electricity, can be restricted represents simultaneously one of the best application fits for POC systems as well as one of the most challenging places to deploy them. Of the many challenges involved in these systems, the preparation and processing of complex samples like stool, vomit, and biopsies are particularly difficult due to the high number and varied nature of mechanical and chemical interferents present in the sample. Previously we have demonstrated the ability to use solar-thermal energy to perform PCR based nucleic acid amplifications. In this work demonstrate how the technique, using similar infrastructure, can also be used to perform solar-thermal based sample processing system for extracting and isolating Vibrio Cholerae nucleic acids from fecal samples. The use of opto-thermal energy enables the use of sunlight to drive thermal lysing reactions in large volumes without the need for external electrical power. Using the system demonstrate the ability to reach a 95°C threshold in less than 5 minutes and maintain a stable sample temperature of +/− 2°C following the ramp up. The system is demonstrated to provide linear results between 104 and 108 CFU/mL when the released nucleic acids were quantified via traditional means. Additionally, we couple the sample processing unit with our previously demonstrated solar-thermal PCR and tablet based detection system to demonstrate very low power sample-in-answer-out detection. PMID:27231636

  10. Solar-thermal complex sample processing for nucleic acid based diagnostics in limited resource settings.

    PubMed

    Gumus, Abdurrahman; Ahsan, Syed; Dogan, Belgin; Jiang, Li; Snodgrass, Ryan; Gardner, Andrea; Lu, Zhengda; Simpson, Kenneth; Erickson, David

    2016-05-01

    The use of point-of-care (POC) devices in limited resource settings where access to commonly used infrastructure, such as water and electricity, can be restricted represents simultaneously one of the best application fits for POC systems as well as one of the most challenging places to deploy them. Of the many challenges involved in these systems, the preparation and processing of complex samples like stool, vomit, and biopsies are particularly difficult due to the high number and varied nature of mechanical and chemical interferents present in the sample. Previously we have demonstrated the ability to use solar-thermal energy to perform PCR based nucleic acid amplifications. In this work demonstrate how the technique, using similar infrastructure, can also be used to perform solar-thermal based sample processing system for extracting and isolating Vibrio Cholerae nucleic acids from fecal samples. The use of opto-thermal energy enables the use of sunlight to drive thermal lysing reactions in large volumes without the need for external electrical power. Using the system demonstrate the ability to reach a 95°C threshold in less than 5 minutes and maintain a stable sample temperature of +/- 2°C following the ramp up. The system is demonstrated to provide linear results between 10(4) and 10(8) CFU/mL when the released nucleic acids were quantified via traditional means. Additionally, we couple the sample processing unit with our previously demonstrated solar-thermal PCR and tablet based detection system to demonstrate very low power sample-in-answer-out detection.

  11. Perfluoroalkyl acids in aqueous samples from Germany and Kenya.

    PubMed

    Shafique, Umer; Schulze, Stefanie; Slawik, Christian; Böhme, Alexander; Paschke, Albrecht; Schüürmann, Gerrit

    2016-06-22

    Continuous monitoring of chemicals in the environment is important to control their fate and to protect human health, flora, and fauna. Perfluoroalkyl acids (PFAAs) have been detected frequently in different environmental compartments during the last 15 years and have drawn much attention because of their environmental persistence, omnipresence, and bioaccumulation potential. Water is an important source of their transport. In the present study, distributions of PFAAs in river water, wastewater treatment plant (WWTP) effluent, and tap water from eastern part of Germany and western part of Kenya were investigated. Eleven perfluorocarboxylic acids (PFCAs) and five perfluorosulfonic acids (PFSAs) were analyzed using liquid chromatography/tandem mass spectrometry. Sum of mean concentrations of eight PFAAs detected in drinking tap water from Leipzig was 11.5 ng L(-1), dominated by perfluorooctanoic acid (PFOA, 6.2 ng L(-1)). Sums of mean riverine concentrations of PFAAs detected in Pleiße/White Elster, Saale, and Elbe (Germany) were 24.8, 54.3, and 26.8 ng L(-1), respectively. Annual flux of PFAAs from River Saale was estimated to be 164 ± 23 kg a(-1). The effluent of WWTP in Halle was found to contain four times higher levels of PFAAs than river water and was dominated by perfluorobutane sulfonate (PFBS) with 32 times higher concentration than the riverine level. It advocates that WWTPs are the point source of contaminating water bodies with PFAAs, and short-chain PFAAs are substituting long-chain homologues. Sums of mean riverine concentrations of PFAAs in Sosiani (Kenya) in samples from sparsely populated and densely populated areas were 58.8 and 109.4 ng L(-1), respectively, indicating that population directly affected the emissions of PFAAs to surface waters. The discussion includes thorough review and comparison of recently published literature reporting occurrence of PFAAs in aqueous matrices. Graphical abstract Perfluoroalkyl acids in aqueous

  12. Sample preparation for thermo-gravimetric determination and thermo-gravimetric characterization of refuse derived fuel.

    PubMed

    Robinson, T; Bronson, B; Gogolek, P; Mehrani, P

    2016-02-01

    Thermo-gravimetric analysis (TGA) is a useful method for characterizing fuels. In the past it has been applied to the study of refuse derived fuel (RDF) and related materials. However, the heterogeneity of RDF makes the preparation of small representative samples very difficult and this difficulty has limited the effectiveness of TGA for characterization of RDF. A TGA method was applied to a variety of materials prepared from a commercially available RDF using a variety of procedures. Applicability of TGA method to the determination of the renewable content of RDF was considered. Cryogenic ball milling was found to be an effective means of preparing RDF samples for TGA. When combined with an effective sample preparation, TGA could be used as an alternative method for assessing the renewable content of RDF.

  13. Searching for Amino Acids in Meteorites and Comet Samples

    NASA Technical Reports Server (NTRS)

    Cook, Jamie Elsila

    2010-01-01

    Chemistry plays an important role in the interdisciplinary field of astrobiology, which strives to understand the origin, distribution, and evolution of life throughout the universe. Chemical techniques are used to search for and characterize the basic ingredients for life, from the elements through simple molecules and up to the more complex compounds that may serve as the ingredients for life. The Astrobiology Analytical Laboratory at NASA Goddard uses state-of-the-art laboratory analytical instrumentation in unconventional ways to examine extraterrestrial materials and tackle some of the big questions in astrobiology. This talk will discuss some of the instrumentation and techniques used for these unique samples, as well as some of our most interesting results. The talk will present two areas of particular interest in our laboratory: (1) the search for chiral excesses in meteoritic amino acids, which may help to explain the origin of homochirality in life on Earth; and (2) the detection of amino acids and amines in material returned by NASA's Stardust mission, which rendevouzed with a cornet and brought back cometary particles to the Earth.

  14. Reducing Spatial Heterogeneity of MALDI Samples with Marangoni Flows During Sample Preparation.

    PubMed

    Lai, Yin-Hung; Cai, Yi-Hong; Lee, Hsun; Ou, Yu-Meng; Hsiao, Chih-Hao; Tsao, Chien-Wei; Chang, Huan-Tsung; Wang, Yi-Sheng

    2016-08-01

    This work demonstrates a method to prepare homogeneous distributions of analytes to improve data reproducibility in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). Natural-air drying processes normally result in unwanted heterogeneous spatial distributions of analytes in MALDI crystals and make quantitative analysis difficult. This study demonstrates that inducing Marangoni flows within drying droplets can significantly reduce the heterogeneity problem. The Marangoni flows are accelerated by changing substrate temperatures to create temperature gradients across droplets. Such hydrodynamic flows are analyzed semi-empirically. Using imaging mass spectrometry, changes of heterogeneity of molecules with the change of substrate temperature during drying processes are demonstrated. The observed heterogeneities of the biomolecules reduce as predicted Marangoni velocities increase. In comparison to conventional methods, drying droplets on a 5 °C substrate while keeping the surroundings at ambient conditions typically reduces the heterogeneity of biomolecular ions by 65%-80%. The observation suggests that decreasing substrate temperature during droplet drying processes is a simple and effective means to reduce analyte heterogeneity for quantitative applications. Graphical Abstract ᅟ.

  15. Preparation and analysis of standardized waste samples for Controlled Ecological Life Support Systems (CELSS)

    NASA Technical Reports Server (NTRS)

    Carden, J. L.; Browner, R.

    1982-01-01

    The preparation and analysis of standardized waste samples for controlled ecological life support systems (CELSS) are considered. Analysis of samples from wet oxidation experiments, the development of ion chromatographic techniques utilizing conventional high pressure liquid chromatography (HPLC) equipment, and an investigation of techniques for interfacing an ion chromatograph (IC) with an inductively coupled plasma optical emission spectrometer (ICPOES) are discussed.

  16. 40 CFR 205.171-2 - Test exhaust system sample selection and preparation.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Test exhaust system sample selection... (CONTINUED) NOISE ABATEMENT PROGRAMS TRANSPORTATION EQUIPMENT NOISE EMISSION CONTROLS Motorcycle Exhaust Systems § 205.171-2 Test exhaust system sample selection and preparation. (a)(1) Exhaust...

  17. Influence of rice sample preparation and milling procedures on milling quality appraisals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this research was to investigate the effect of sample preparation and milling procedure on milling quality appraisals of rough rice. Samples of freshly harvested medium-grain rice (M202) with different initial moisture contents (MCs) ranging from 20.2% to 25.1% (w.b.) were used for...

  18. Glass sample preparation and performance investigations. [solar x-ray imager

    NASA Technical Reports Server (NTRS)

    Johnson, R. Barry

    1992-01-01

    This final report details the work performed under this delivery order from April 1991 through April 1992. The currently available capabilities for integrated optical performance modeling at MSFC for large and complex systems such as AXAF were investigated. The Integrated Structural Modeling (ISM) program developed by Boeing for the U.S. Air Force was obtained and installed on two DECstations 5000 at MSFC. The structural, thermal and optical analysis programs available in ISM were evaluated. As part of the optomechanical engineering activities, technical support was provided in the design of support structure, mirror assembly, filter wheel assembly and material selection for the Solar X-ray Imager (SXI) program. As part of the fabrication activities, a large number of zerodur glass samples were prepared in different sizes and shapes for acid etching, coating and polishing experiments to characterize the subsurface damage and stresses produced by the grinding and polishing operations. Various optical components for AXAF video microscope and the x-ray test facility were also fabricated. A number of glass fabrication and test instruments such as a scatter plate interferometer, a gravity feed saw and some phenolic cutting blades were fabricated, integrated and tested.

  19. A combinatorial sample-preparation robot system using the volumetric-weighing method

    NASA Astrophysics Data System (ADS)

    Suehara, S.; Konishi, T.; Fujimoto, K.; Takeda, T.; Fukuda, M.; Koike, M.; Inoue, S.; Watanabe, M.

    2006-01-01

    We have developed a new automatic sample-preparation robot system with use of the volumetric-weighing method. In this system, slurries, aqueous solutions, and other wet reagents are employed as starting materials and 64 ( 8×8) samples at the maximum are prepared on a library plate of 35 mm × 35 mm size. Volumetric-weighing and mixing of the starting materials and distributing reaction mixtures to the library plate are automatically performed by computer-controlled mechanisms with an easy-to-use programming software interface. While this robot is designed in terms of space saving and portability, it is able to equip with an atmosphere-controlled furnace to sinter the samples on the library plate. Typical preparation time for a library plate of 36 ( 6×6) samples is less than 40 min. This robot system is promising in enhancing throughput of wet-chemically synthesized materials researches.

  20. Preparation and characterization of microporous fibers for sample preparation and LC-MS determination of drugs.

    PubMed

    Buszewski, Boguslaw; Nowaczyk, Jacek; Ligor, Tomasz; Olszowy, Pawel; Ligor, Magdalena; Wasiniak, Bartlomiej; Miekisch, Wolfram; Schubert, Jochen K; Amann, Anton

    2009-07-01

    The aim of this study was the preparation of polypyrrole (PPy) fibers for solid phase microextraction (SPME). PPy coatings were obtained during the electrochemical polymerization process. The utility of various metal wires (Fe, Cu, Ag, Cu/Ag, kanthal and medical stainless steel) as a support for polymers was compared. Various experimental conditions of the synthesis process such as scan rate, voltage limits and number of scans and deposition time were applied. The average polymer thickness was in the range of 7-125 microm and its weight was in the scope of 0.65-5.6 mg. Different techniques, mainly elemental analysis, Fourier transform infrared spectroscopy, microscopy, and chromatography were performed for the characterization of obtained fibers with microporous structure. The extraction efficiency of cardiovascular drugs (metoprolol, propranolol, oxprenolol, propafenone and mexiletine) by means of fibers was tested. The concentration of mentioned compounds in standard solution was in the span of 10-150 ng/mL. LC-MS was employed for determination of drugs in desorption solution. LODs varied from 0.013 to 1.51 ng/mL for metoprolol and mexiletine respectively. The repeatability of extraction was obtained with the RSD values lower than 10%.

  1. Impact of sample preparation on mineralogical analysis of zero-valent iron reactive barrier materials

    SciTech Connect

    Phillips, Debra Helen; Gu, Baohua; Watson, David B; Roh, Yul

    2003-03-01

    Permeable reactive barriers (PRBs) of zero-valent iron (Fe{sup 0}) are increasingly being used to remediate contaminated ground water. Corrosion of Fe{sup 0} filings and the formation of precipitates can occur when the PRB material comes in contact with ground water and may reduce the lifespan and effectiveness of the barrier. At present, there are no routine procedures for preparing and analyzing the mineral precipitates from Fe{sup 0} PRB material. These procedures are needed because mineralogical composition of corrosion products used to interpret the barrier processes can change with iron oxidation and sample preparation. The objectives of this study were (i) to investigate a method of preparing Fe{sup 0} reactive barrier material for mineralogical analysis by X-ray diffraction (XRD), and (ii) to identify Fe mineral phases and rates of transformations induced by different mineralogical preparation techniques. Materials from an in situ Fe{sup 0} PRB were collected by undisturbed coring and processed for XRD analysis after different times since sampling for three size fractions and by various drying treatments. We found that whole-sample preparation for analysis was necessary because mineral precipitates occurred within the PRB material in different size fractions of the samples. Green rusts quickly disappeared from acetone-dried samples and were not present in air-dried and oven-dried samples. Maghemite/magnetite content increased over time and in oven-dried samples, especially after heating to 105 C. We conclude that care must be taken during sample preparation of Fe{sup 0} PRB material, especially for detection of green rusts, to ensure accurate identification of minerals present within the barrier system.

  2. Design and construction of a sample preparation chamber for atomic beam scattering

    SciTech Connect

    Nielsen, C.

    1992-05-18

    A new type of atomic beam scattering spectrometer was built to advance the usefulness of the atomic beam scattering technique as a surface dynamics probe. The facility was not only built to investigate the typical alkali halide samples such as NaCl, NaF, and LiF, but also to investigate metallic surfaces. Metal samples are more complicated to study, due to their reactive surfaces and the sample preparation process. A surface analysis chamber was constructed as an attachment to the scattering facility to treat samples under ultra high vacuum (UHV) and then transfer these samples into the scattering facility. This surface analysis chamber is referred to as the sample preparation chamber and is the basis for this thesis.

  3. Pilot study of a rapid and minimally instrumented sputum sample preparation method for molecular diagnosis of tuberculosis

    PubMed Central

    Ferguson, Tanya M.; Weigel, Kris M.; Lakey Becker, Annie; Ontengco, Delia; Narita, Masahiro; Tolstorukov, Ilya; Doebler, Robert; Cangelosi, Gerard A.; Niemz, Angelika

    2016-01-01

    Nucleic acid amplification testing (NAAT) enables rapid and sensitive diagnosis of tuberculosis (TB), which facilitates treatment and mitigates transmission. Nucleic acid extraction from sputum constitutes the greatest technical challenge in TB NAAT for near-patient settings. This report presents preliminary data for a semi-automated sample processing method, wherein sputum is disinfected and liquefied, followed by PureLyse® mechanical lysis and solid-phase nucleic acid extraction in a miniaturized, battery-operated bead blender. Sputum liquefaction and disinfection enabled a >104 fold reduction in viable load of cultured Mycobacterium tuberculosis (M.tb) spiked into human sputum, which mitigates biohazard concerns. Sample preparation via the PureLyse® method and a clinically validated manual method enabled positive PCR-based detection for sputum spiked with 104 and 105 colony forming units (cfu)/mL M.tb. At 103 cfu/mL sputum, four of six and two of six samples amplified using the comparator and PureLyse® method, respectively. For clinical specimens from TB cases and controls, the two methods provided 100% concordant results for samples with 1 mL input volume (N = 41). The semi-automated PureLyse® method therefore performed similarly to a validated manual comparator method, but is faster, minimally instrumented, and can be integrated into TB molecular diagnostic platforms designed for near-patient low-resource settings. PMID:26785769

  4. Effects of packaging and storage conditions on the quality of amoxicillin-clavulanic acid – an analysis of Cambodian samples

    PubMed Central

    2013-01-01

    Background The use of substandard and degraded medicines is a major public health problem in developing countries such as Cambodia. A collaborative study was conducted to evaluate the quality of amoxicillin–clavulanic acid preparations under tropical conditions in a developing country. Methods Amoxicillin-clavulanic acid tablets were obtained from outlets in Cambodia. Packaging condition, printed information, and other sources of information were examined. The samples were tested for quantity, content uniformity, and dissolution. Authenticity was verified with manufacturers and regulatory authorities. Results A total of 59 samples were collected from 48 medicine outlets. Most (93.2%) of the samples were of foreign origin. Using predetermined acceptance criteria, 12 samples (20.3%) were non-compliant. Eight (13.6%), 10 (16.9%), and 20 (33.9%) samples failed quantity, content uniformity, and dissolution tests, respectively. Samples that violated our observational acceptance criteria were significantly more likely to fail the quality tests (Fisher’s exact test, p < 0.05). Conclusions Improper packaging and storage conditions may reduce the quality of amoxicillin–clavulanic acid preparations at community pharmacies. Strict quality control measures are urgently needed to maintain the quality of amoxicillin–clavulanic acid in tropical countries. PMID:23773420

  5. Detection of genetically modified crops and their derivatives: critical steps in sample preparation and extraction.

    PubMed

    Terry, Catherine F; Harris, Neil; Parkes, Helen C

    2002-01-01

    The detection of genetically modified crops in foodstuff relies on detection of transgenic DNA or protein material in the sample matrix. Purified DNA or proteins are used as analytical material for polymerase chain reaction technologies and immunodiagnostics. Successful sample preparation is critical to the validity of subsequent analysis. For routine analysis, a good sample preparation technique should be simple, safe, and inexpensive while reproducibly generating DNA/protein of sufficient quality and yield. The suitability of isolated DNA or protein as an analyte for a detection or characterization technique depends on amount or concentration, purity, and integrity, each of which may be influenced by sample matrix and the extraction technique, and, in turn, may impact the validity of analytical techniques. The key sample preparation steps of homogenization, pretreatment, extraction, and purification are discussed as well as typical analytical methods. Consideration is given to application of these steps for particular sample matrixes to maximize yield, reduce inhibition effects, and minimize contamination. The choice of the most appropriate and valid methods for sample preparation from particular foods is discussed with respect to DNA analysis. Attention is also given to ease of use, cost, and generic applicability of the procedures.

  6. Preparation of protein samples for mass spectrometry and N-terminal sequencing.

    PubMed

    Glenn, Gary

    2014-01-01

    The preparation of protein samples for mass spectrometry and N-terminal sequencing is a key step in successfully identifying proteins. Mass spectrometry is a very sensitive technique, and as such, samples must be prepared carefully since they can be subject to contamination of the sample (e.g., due to incomplete subcellular fractionation or purification of a multiprotein complex), overwhelming of the sample by highly abundant proteins, and contamination from skin or hair (keratin can be a very common hit). One goal of sample preparation for mass spec is to reduce the complexity of the sample - in the example presented here, mitochondria are purified, solubilized, and fractionated by sucrose density gradient sedimentation prior to preparative 1D SDS-PAGE. It is important to verify the purity and integrity of the sample so that you can have confidence in the hits obtained. More protein is needed for N-terminal sequencing and ideally it should be purified to a single band when run on an SDS-polyacrylamide gel. The example presented here involves stably expressing a tagged protein in HEK293 cells and then isolating the protein by affinity purification and SDS-PAGE.

  7. TruSeq Stranded mRNA and Total RNA Sample Preparation Kits

    Cancer.gov

    Total RNA-Seq enabled by ribosomal RNA (rRNA) reduction is compatible with formalin-fixed paraffin embedded (FFPE) samples, which contain potentially critical biological information. The family of TruSeq Stranded Total RNA sample preparation kits provides a unique combination of unmatched data quality for both mRNA and whole-transcriptome analyses, robust interrogation of both standard and low-quality samples and workflows compatible with a wide range of study designs.

  8. Preparation of Mg-MgH2 flakes by planetary ball milling with stearic acid and their hydrogen storage properties

    NASA Astrophysics Data System (ADS)

    Hong, Seong-Hyeon; Song, Myoung Youp

    2016-05-01

    Many studies preparing magnesium hydride using catalyst addition were performed, resulting in the preparation of additive-containing magnesium hydride. Preparation of a sample with a MgH2 phase without additives requires high pressure and high temperature and is time-demanding. In order to prepare an additive-free sample with a MgH2 phase, 90 wt% Mg+10 wt% MgH2 (named 90Mg+10MgH2) was milled under a hydrogen atmosphere with 6 wt% stearic acid as a process-controlling agent, which led to a formation of Mg-MgH2 flakes. The hydrogen storing and releasing properties of the prepared flakes were investigated and compared with those of purchased MgH2. A sample with a majority fraction of MgH2 phase was prepared by planetary ball milling of 90 Mg+10 MgH2 with 6 wt% stearic acid. The resultant particles of 90 Mg+10 MgH2 obtained after hydridingdehydriding cycling were much smaller and had significantly more cracks and defects than those of MgH2 after hydriding-dehydriding cycling. 90 Mg+10 MgH2 released 0.12 wt% hydrogen for 4 min, 3.70 wt% for 20 min, and 5.30 wt% for 60 min at 648 K at the first cycle.

  9. Preparation of Conjugated Linoleic Acid Microemulsions and their Biodistribution.

    PubMed

    Kishita, Kazuaki; Ibaraki, Kayo; Itakura, Shoko; Yamasaki, Yumi; Nishikata, Naoko; Yamamoto, Kenji; Shimizu, Masataka; Nishiyama, Kazuo; Yamasaki, Masao

    2016-01-01

    Conjugated linoleic acid (CLA) has several beneficial biological properties. Specifically, trans10, cis12-CLA, one of the CLA isomers, has strong physiologic activity against cancer and obesity. However, compared with cis9, trans11-CLA, a naturally occurring CLA isomer, trans10, cis12-CLA tends to be easily metabolized. Therefore, to make efficient use of its biological properties, it is necessary to overcome the rapid clearance of trans10, cis12-CLA from the blood. Here, we employed premix membrane emulsification to prepare two oil-in-water CLA microemulsions (CLA-ME), 100 nm CLA-ME and 200 nm CLA-ME, and investigated their pharmacokinetics in a mouse model. We report that 100 nm CLA-ME contributed to the concentration of blood CLA for longer than 200 nm CLA-ME, indicating that small CLA microparticles were more suitable for maintaining blood trans10, cis12-CLA levels in vivo. However, both CLA-ME could be hardly detected in blood and other tissues 24 h after administration, suggesting that additional strategies for prolonging CLA-ME half-life are required.

  10. Preparation of alginate beads containing a prodrug of diethylenetriaminepentaacetic acid

    PubMed Central

    Yang, Yu-Tsai; Di Pasqua, Anthony J.; He, Weiling; Tsai, Tsuimin; Sueda, Katsuhiko; Zhang, Yong; Jay, Michael

    2012-01-01

    A penta-ethyl ester prodrug of the radionuclide decorporation agent diethylenetriaminepentaacetic acid (DTPA), which exists as an oily liquid, was encapsulated in alginate beads by the ionotropic gelation method. An optimal formulation was found by varying initial concentrations of DTPA pentaethyl ester, alginate polymer, Tween 80 surfactant and calcium chloride. All prepared alginate beads were ~1.6 mm in diameter, and the optimal formulation had loading and encapsulation efficiencies of 91.0 ± 1.1 and 72.6 ± 2.2%, respectively, and only 3.2 ± 0.8% water absorption after storage at room temperature in ~80% relative humidity. Moreover, Fourier transform infrared spectroscopy showed that DTPA penta-ethyl ester did not react with excipients during formation of the DTPA penta-ethyl ester-containing alginate beads. Release of prodrug from alginate beads was via anomalous transport, and its stability enhanced by encapsulation. Collectively, these data suggest that this solid dosage form may be suitable for oral administration after radionuclide contamination. PMID:23399237

  11. An efficient and cost-effective method for preparing transmission electron microscopy samples from powders

    DOE PAGES

    Wen, Haiming; Lin, Yaojun; Seidman, David N.; ...

    2015-09-09

    The preparation of transmission electron microcopy (TEM) samples from powders with particle sizes larger than ~100 nm poses a challenge. The existing methods are complicated and expensive, or have a low probability of success. Herein, we report a modified methodology for preparation of TEM samples from powders, which is efficient, cost-effective, and easy to perform. This method involves mixing powders with an epoxy on a piece of weighing paper, curing the powder–epoxy mixture to form a bulk material, grinding the bulk to obtain a thin foil, punching TEM discs from the foil, dimpling the discs, and ion milling the dimpledmore » discs to electron transparency. Compared with the well established and robust grinding–dimpling–ion-milling method for TEM sample preparation for bulk materials, our modified approach for preparing TEM samples from powders only requires two additional simple steps. In this article, step-by-step procedures for our methodology are described in detail, and important strategies to ensure success are elucidated. Furthermore, our methodology has been applied successfully for preparing TEM samples with large thin areas and high quality for many different mechanically milled metallic powders.« less

  12. An efficient and cost-effective method for preparing transmission electron microscopy samples from powders

    SciTech Connect

    Wen, Haiming; Lin, Yaojun; Seidman, David N.; Schoenung, Julie M.; van Rooyen, Isabella J.; Lavernia, Enrique J.

    2015-09-09

    The preparation of transmission electron microcopy (TEM) samples from powders with particle sizes larger than ~100 nm poses a challenge. The existing methods are complicated and expensive, or have a low probability of success. Herein, we report a modified methodology for preparation of TEM samples from powders, which is efficient, cost-effective, and easy to perform. This method involves mixing powders with an epoxy on a piece of weighing paper, curing the powder–epoxy mixture to form a bulk material, grinding the bulk to obtain a thin foil, punching TEM discs from the foil, dimpling the discs, and ion milling the dimpled discs to electron transparency. Compared with the well established and robust grinding–dimpling–ion-milling method for TEM sample preparation for bulk materials, our modified approach for preparing TEM samples from powders only requires two additional simple steps. In this article, step-by-step procedures for our methodology are described in detail, and important strategies to ensure success are elucidated. Furthermore, our methodology has been applied successfully for preparing TEM samples with large thin areas and high quality for many different mechanically milled metallic powders.

  13. Focused ion beam preparation of samples for X-ray nanotomography.

    PubMed

    Lombardo, Jeffrey J; Ristau, Roger A; Harris, William M; Chiu, Wilson K S

    2012-09-01

    The preparation of hard material samples with the necessary size and shape is critical to successful material analysis. X-ray nanotomography requires that samples are sufficiently thin for X-rays to pass through the sample during rotation for tomography. One method for producing samples that fit the criteria for X-ray nanotomography is focused ion beam/scanning electron microscopy (FIB/SEM) which uses a focused beam of ions to selectively mill around a region of interest and then utilizes a micromanipulator to remove the milled-out sample from the bulk material and mount it on a sample holder. In this article the process for preparing X-ray nanotomography samples in multiple shapes and sizes is discussed. Additionally, solid-oxide fuel cell anode samples prepared through the FIB/SEM technique underwent volume-independence studies for multiple properties such as volume fraction, average particle size, tortuosity and contiguity to observe the characteristics of FIB/SEM samples in X-ray nanotomography.

  14. Surface Cleaning Techniques: Ultra-Trace ICP-MS Sample Preparation and Assay of HDPE

    SciTech Connect

    Overman, Nicole R.; Hoppe, Eric W.; Addleman, Raymond S.

    2013-06-01

    The world’s most sensitive radiation detection and assay systems depend upon ultra-low background (ULB) materials to reduce unwanted radiological backgrounds. Herein, we evaluate methods to clean HDPE, a material of interest to ULB systems and the means to provide rapid assay of surface and bulk contamination. ULB level material and ultra-trace level detection of actinide elements is difficult to attain, due to the introduction of contamination from sample preparation equipment such as pipette tips, sample vials, forceps, etc. and airborne particulate. To date, literature available on the cleaning of such polymeric materials and equipment for ULB applications and ultra-trace analyses is limited. For these reasons, a study has been performed to identify an effective way to remove surface contamination from polymers in an effort to provide improved instrumental detection limits. Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) was utilized to assess the effectiveness of a variety of leachate solutions for removal of inorganic uranium and thorium surface contamination from polymers, specifically high density polyethylene (HDPE). HDPE leaching procedures were tested to optimize contaminant removal of thorium and uranium. Calibration curves for thorium and uranium ranged from 15 ppq (fg/mL) to 1 ppt (pg/mL). Detection limits were calculated at 6 ppq for uranium and 7 ppq for thorium. Results showed the most effective leaching reagent to be clean 6 M nitric acid for 72 hour exposures. Contamination levels for uranium and thorium found in the leachate solutions were significant for ultralow level radiation detection applications.

  15. Accumulated analyses of amino acid precursors in returned lunar samples

    NASA Technical Reports Server (NTRS)

    Fox, S. W.; Harada, K.; Hare, P. E.

    1973-01-01

    Six amino acids (glycine, alanine, aspartic acid, glutamic acid, serine, and threonine) obtained by hydrolysis of extracts have been quantitatively determined in ten collections of fines from five Apollo missions. Although the amounts found, 7-45 ng/g, are small, the lunar amino acid/carbon ratios are comparable to those of the carbonaceous chondrites, Murchison and Murray, as analyzed by the same procedures. Since both the ratios of amino acid to carbon, and the four or five most common types of proteinous amino acid found, are comparable for the two extraterrestrial sources despite different cosmophysical histories of the moon and meteorites, common cosmochemical processes are suggested.

  16. Sample preparation of energy materials for X-ray nanotomography with micromanipulation.

    PubMed

    Chen-Wiegart, Yu-chen Karen; Camino, Fernando E; Wang, Jun

    2014-06-06

    X-ray nanotomography presents an unprecedented opportunity to study energy storage/conversion materials at nanometer scales in three dimensions, with both elemental and chemical sensitivity. A critical step in obtaining high-quality X-ray nanotomography data is reliable sample preparation to ensure that the entire sample fits within the field of view of the X-ray microscope. Although focused-ion-beam lift-out has previously been used for large sample (few to tens of microns) preparation, a difficult undercut and lift-out procedure results in a time-consuming sample preparation process. Herein, we propose a much simpler and direct sample preparation method to resolve the issues that block the view of the sample base after milling and during the lift-out process. This method is applied on a solid-oxide fuel cell and a lithium-ion battery electrode, before numerous critical 3D morphological parameters are extracted, which are highly relevant to their electrochemical performance. A broad application of this method for microstructure study with X-ray nanotomography is discussed and presented.

  17. High-throughput automated microfluidic sample preparation for accurate microbial genomics

    PubMed Central

    Kim, Soohong; De Jonghe, Joachim; Kulesa, Anthony B.; Feldman, David; Vatanen, Tommi; Bhattacharyya, Roby P.; Berdy, Brittany; Gomez, James; Nolan, Jill; Epstein, Slava; Blainey, Paul C.

    2017-01-01

    Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbitrary numbers of reaction and clean-up or capture steps. By reducing the sample quantity requirements, we enabled low-input (∼10,000 cells) whole-genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil micro-colonies with superior results. We also leveraged the enhanced throughput to sequence ∼400 clinical Pseudomonas aeruginosa libraries and demonstrate excellent single-nucleotide polymorphism detection performance that explained phenotypically observed antibiotic resistance. Fully-integrated lab-on-chip sample preparation overcomes technical barriers to enable broader deployment of genomics across many basic research and translational applications. PMID:28128213

  18. Two-in-one sample preparation for plan-view TEM.

    PubMed

    Sáfrán, György; Szász, Noémi; Sáfrán, Eszter

    2015-07-01

    Transmission electron microscopy (TEM) sample preparation requires special skills, it is time consuming and costly, hence, an increase of the efficiency is of primary importance. This article describes a method that duplicates the yield of the conventional mechanical and ion beam preparation of plan-view TEM samples. As a modification of the usual procedures, instead of one two different samples are comprised in a single specimen. The two pre-cut slabs, one from each samples, are embedded side by side in the window of a 3 mm dia Ti disk and the specimen is thinned mechanically and by ion milling until perforation that occurs at the interface of the two different slabs. That, with proper implementation, provides acceptable size thin area for the TEM study of both samples. The suitability of the two-in-one method has been confirmed through examples.

  19. [Preparation of samples for proficiency testing of pesticide residue analysis in processed foods].

    PubMed

    Okihashi, Masahiro; Osakada, Masakazu; Uchida, Kotaro; Nagayoshi, Haruna; Yamaguchi, Takahiro; Kakimoto, Kensaku; Nakayama, Yukiko; Obana, Hirotaka

    2010-01-01

    To conduct proficiency testing for the analysis of pesticide residues in processed foods, fortified samples of retort curry and pancake were examined. In the case of retort curry, heating and mixing were necessary at the time of preparation to provide a homogenous analytical sample. A mixture of 4 carbamates and 11 organophosphorus pesticides was spiked and 14 of them showed consistent results in the samples. In the case of pancake, 10 kinds of pesticides were added to the pastry. The prepared pastry was them cooked. The relative concentrations of most of the pesticides in the pancake were not affected and all the pesticides showed consistent results in the samples. These results showed that the two tested samples were suitable for proficiency testing.

  20. A review of toxicity from topical salicylic acid preparations.

    PubMed

    Madan, Raman K; Levitt, Jacob

    2014-04-01

    Topical salicylic acid is often used in dermatologic conditions because of its keratolytic, bacteriostatic, fungicidal, and photoprotective properties. The bioavailability of salicylic acid differs depending on the vehicle used and pH of transcellular fluids. Although rare, salicylic acid toxicity (salicylism) can occur from topical application. Physicians should be mindful of the potential for salicylism or even death from topically applied salicylic acid.

  1. Electric transport measurements on bulk, polycrystalline MgB2 samples prepared at various reaction temperatures

    NASA Astrophysics Data System (ADS)

    Wiederhold, A.; Koblischka, M. R.; Inoue, K.; Muralidhar, M.; Murakami, M.; Hartmann, U.

    2016-03-01

    A series of disk-shaped, bulk MgB2 superconductors (sample diameter up to 4 cm) was prepared in order to improve the performance for superconducting super-magnets. Several samples were fabricated using a solid state reaction in pure Ar atmosphere from 750 to 950oC in order to determine the optimum processing parameters to obtain the highest critical current density as well as large trapped field values. Additional samples were prepared with added silver (up to 10 wt.-%) to the Mg and B powder. Magneto-resistance data and I/V-characteristics were recorded using an Oxford Instruments Teslatron system. From Arrhenius plots, we determine the TAFF pinning potential, U 0. The I/V-characteristics yield detailed information on the current flow through the polycrystalline samples. The current flow is influenced by the presence of pores in the samples. Our analysis of the achieved critical currents together with a thorough microstructure investigation reveals that the samples prepared at temperatures between 775°C and 805°C exhibit the smallest grains and the best connectivity between them, while the samples fabricated at higher reaction temperatures show a reduced connectivity and lower pinning potential. Doping the samples with silver leads to a considerable increase of the pinning potential and hence, the critical current densities.

  2. A Simple Sample Preparation with HPLC-UV Method for Estimation of Clomipramine from Plasma

    PubMed Central

    Mostafavi, Sayed Abolfazl; Tahvilian, Reza; Dehghani Poudeh, Masoumeh; Rafeepour, Zeinab

    2010-01-01

    Clomipramine is a tricyclic antidepressant. Different methods for determination of clomipramine hydrochloride in plasma have been described. Most of these procedures favor the use of acidic back-extraction in extraction procedure and HPLC as the analytical technique. In this study, the clomipramine extraction procedure was modified and a direct injection to the column was performed to shorten the time of sample preparation considerably. Furthermore, the method was applied in bioequivalence study of new formulations of clomipramine in comparison with reference tablets. The drug and internal standard were extracted from plasma with heptan : isoamyl alcohol (95:5) and re-extracted with 200 μL of orthophosphoric acid (0.3% v/v). The organic layer was discharged and analysis was performed on C8 reverse phase ODS2 HPLC column with a mobile phase, acetonitrile : water (75:25) and UV detection set at 215 nm. Additionally, a single dose study was carried out with a two-sequence, crossover block-randomized design for bioequivalence study. Clomipramine tablets (3 × 25 mg) of either formulations (reference or test products) were administered separately in two occasions to 12 fasting healthy male volunteers. Blood samples were taken prior to and at 9 points within 48 h after dose administration. The retention time of internal standard (cisapride), clomipramine, and desmethyl clomipramine were 5.6 ± 0.2, 10.3 ± 0.3, and 9.5 ± 0.3 min, respectively. The standard curve covering the concentration ranges of 2.5-120 ng/mL was linear (r2 = 0.9950 and 0.9979) for clomipramine and desmethyl clomipramine. The co-efficient of variation for intra-day and inter-day accuracy and precision was less than 18.3%. The pharmacokinetic parameters Cmax and Tmax were obtained directly from plasma clomipramine concentrations. Kel was estimated by log-linear regression and AUC was calculated by the linear trapezoidal rule. The pharmacokinetic parameters AUC and Cmax were tested for equivalence

  3. Specific features of sample preparation from amorphous aluminum alloys for transmission electron microscopy

    SciTech Connect

    Volkov, P. A.; Todorova, E. V.; Bakhteeva, N. D.; Ivanova, A. G.; Vasil'ev, A. L.

    2011-05-15

    An aluminum amorphous alloy doped with transition (Fe and Ni) and rare earth (La) metals has been used as an object of systematic study of the structural transformations that are characteristic of different methods of sample preparation for transmission electron microscopy (the mechanical tearing of ribbons, electrochemical thinning, and Ar{sup +}-ion etching under different conditions). The results of X-ray diffraction analysis and a calorimetric study of the structure in comparison with electron microscopy data made it possible to determine the optimal method of sample preparation, which ensures minimum distortions in the structure of metastable amorphous alloys with a low crystallization temperature.

  4. Open focused microwave-assisted sample preparation for rapid total and mercury species determination in environmental solid samples.

    PubMed

    Tseng, C M; Garraud, H; Amouroux, D; Donard, O F; de Diego, A

    1998-01-01

    This paper describes rapid, simple microwave-assisted leaching/ digestion procedures for total and mercury species determination in sediment samples and biomaterials. An open focused microwave system allowed the sample preparation time to be dramatically reduced to only 24 min when a power of 40-80 W was applied. Quantitative leaching of methylmercury from sediments by HNO(3) solution and complete dissolution of biomaterials by an alkaline solution, such as 25% TMAH solution, were obtained. Methylmercury compounds were kept intact without decomposition or losses by evaporation. Quantitative recoveries of total mercury were achieved with a two-step microwave attack using a combination of HNO(3) and H(2)0(2) solutions as extractant. The whole pretreatment procedure only takes 15 min, which can be further shortened by an automated robust operation with an open focused system. These analytical procedures were validated by the analysis of environmental certified reference materials. The results confirm that the open focused microwave technique is a promising tool for solid sample preparation in analytical and environmental chemistry.

  5. Effect of Oral Coadministration of Ascorbic Acid with Ling Zhi Preparation on Pharmacokinetics of Ganoderic Acid A in Healthy Male Subjects: A Randomized Crossover Study

    PubMed Central

    Tawasri, Patcharanee; Ampasavate, Chadarat; Tharatha, Somsak

    2016-01-01

    The objective of this randomized, open-label, single-dose, two-phase crossover study was to determine the effect of ascorbic acid on pharmacokinetics of ganoderic acid A, an important biologically active triterpenoid compound with anticancer activities, following oral administration of water extract of fruiting bodies of Ling Zhi in 12 healthy male subjects. Each subject was randomized to receive either one of the two regimens: (1) a single dose of 3,000 mg of the Ling Zhi preparation or (2) a single dose of 3,000 mg of the Ling Zhi preparation in combination with 2,500 mg of ascorbic acid. After a washout period of at least two weeks, subjects were switched to receive the alternate regimen. Blood samples were collected in each phase immediately before dosing and at specific time points for 8 hours after dosing. Plasma ganoderic acid A concentrations were quantified using liquid chromatography-mass spectrometry (LC-MS). The pharmacokinetic parameters analyzed were maximal plasma concentration (Cmax), time to reach peak concentration (Tmax), area under the plasma concentration-time curve (AUC), and half-life (t1/2). An oral coadministration of ascorbic acid with Ling Zhi preparation did not significantly alter the pharmacokinetic parameters of ganoderic acid A in healthy male subjects. PMID:27747224

  6. Instrument and method for X-ray diffraction, fluorescence, and crystal texture analysis without sample preparation

    NASA Technical Reports Server (NTRS)

    Gendreau, Keith (Inventor); Martins, Jose Vanderlei (Inventor); Arzoumanian, Zaven (Inventor)

    2010-01-01

    An X-ray diffraction and X-ray fluorescence instrument for analyzing samples having no sample preparation includes a X-ray source configured to output a collimated X-ray beam comprising a continuum spectrum of X-rays to a predetermined coordinate and a photon-counting X-ray imaging spectrometer disposed to receive X-rays output from an unprepared sample disposed at the predetermined coordinate upon exposure of the unprepared sample to the collimated X-ray beam. The X-ray source and the photon-counting X-ray imaging spectrometer are arranged in a reflection geometry relative to the predetermined coordinate.

  7. Process for the preparation of 3,4-dihydroxybutanoic acid and salts thereof

    DOEpatents

    Hollingsworth, Rawle I.

    1994-01-01

    A process for the preparation of 3,4-dihydroxybutanoic acid (1) and salts thereof from a glucose source containing 1,4-linked glucose as a substituent is described. The process uses an alkali metal hdyroxide and hydrogen peroxide to convert the glucose source to (1). The compound (1) is useful as a chemical intermediate to naturally occurring fatty acids and is used to prepare 3,4-dihydroxybutanoic acid-gamma-lactone (2) and furanone (3), particularly stereoisomers of these compounds.

  8. Process For The Preparation Of 3,4-Dihyd Roxybutanoic Acid And Salts Thereof

    DOEpatents

    Hollingsworth, Rawle I.

    1994-06-07

    A process for the preparation of 3,4-dihydroxybutanoic acid (1) and salts thereof from a glucose source containing 1,4-linked glucose as a substituent is described. The process uses an alkali metal hdyroxide and hydrogen peroxide to convert the glucose source to (1). The compound (1) is useful as a chemical intermediate to naturally occurring fatty acids and is used to prepare 3,4-dihydroxybutanoic acid-gamma-lactone (2) and furanone (3), particularly stereoisomers of these compounds.

  9. Recent advances in sample preparation techniques and methods of sulfonamides detection - A review.

    PubMed

    Dmitrienko, Stanislava G; Kochuk, Elena V; Apyari, Vladimir V; Tolmacheva, Veronika V; Zolotov, Yury A

    2014-11-19

    Sulfonamides (SAs) have been the most widely used antimicrobial drugs for more than 70 years, and their residues in foodstuffs and environmental samples pose serious health hazards. For this reason, sensitive and specific methods for the quantification of these compounds in numerous matrices have been developed. This review intends to provide an updated overview of the recent trends over the past five years in sample preparation techniques and methods for detecting SAs. Examples of the sample preparation techniques, including liquid-liquid and solid-phase extraction, dispersive liquid-liquid microextraction and QuEChERS, are given. Different methods of detecting the SAs present in food and feed and in environmental, pharmaceutical and biological samples are discussed.

  10. Development and Evaluation of a Micro- and Nanoscale Proteomic Sample Preparation Method

    SciTech Connect

    Wang, Haixing H.; Qian, Weijun; Mottaz, Heather M.; Clauss, Therese R.W.; Anderson, David J.; Moore, Ronald J.; Camp, David G.; Khan, Arshad H.; Sforza, Daniel M.; Pallavicini, Maria; Smith, Desmond J.; Smith, Richard D.

    2005-10-05

    Efficient and effective sample preparation of micro- and nano-scale (micro- and nano-gram) clinical specimens for proteomic applications is often difficult due to losses during the processing steps. Herein we describe a simple “single-tube” preparation protocol appropriate for small proteomic samples using the organic co-solvent, trifluoroethanol (TFE). TFE facilitates both protein extraction and protein denaturation without requiring a separate cleanup step, thus minimizing sample loss. The performance of the TFE method was initially evaluated by comparing to traditional detergent-based methods on relatively large scale sample processing using human breast cancer cells and mouse brain tissue. The results demonstrated that the TFE protocol provided comparable results to the traditional detergent-based protocols for larger samples (milligrams), based on both sample recovery and peptide/protein identification. The effectiveness of this protocol for micro- and nano-scale sample processing was then evaluated for the extraction of proteins/peptides and shown effective for small mouse brain tissue samples (~ 20 μg total protein content) and also for samples of ~ 5 000 human breast cancer MCF-7 cells (~ 500 ng total protein content), where the detergent-based methods were ineffective due to losses during cleanup and transfer steps.

  11. Author Contribution to the Pu Handbook II: Chapter 37 LLNL Integrated Sample Preparation Glovebox (TEM) Section

    SciTech Connect

    Wall, Mark A.

    2016-10-25

    The development of our Integrated Actinide Sample Preparation Laboratory (IASPL) commenced in 1998 driven by the need to perform transmission electron microscopy studies on naturally aged plutonium and it’s alloys looking for the microstructural effects of the radiological decay process (1). Remodeling and construction of a laboratory within the Chemistry and Materials Science Directorate facilities at LLNL was required to turn a standard radiological laboratory into a Radiological Materials Area (RMA) and Radiological Buffer Area (RBA) containing type I, II and III workplaces. Two inert atmosphere dry-train glove boxes with antechambers and entry/exit fumehoods (Figure 1), having a baseline atmosphere of 1 ppm oxygen and 1 ppm water vapor, a utility fumehood and a portable, and a third double-walled enclosure have been installed and commissioned. These capabilities, along with highly trained technical staff, facilitate the safe operation of sample preparation processes and instrumentation, and sample handling while minimizing oxidation or corrosion of the plutonium. In addition, we are currently developing the capability to safely transfer small metallographically prepared samples to a mini-SEM for microstructural imaging and chemical analysis. The gloveboxes continue to be the most crucial element of the laboratory allowing nearly oxide-free sample preparation for a wide variety of LLNL-based characterization experiments, which includes transmission electron microscopy, electron energy loss spectroscopy, optical microscopy, electrical resistivity, ion implantation, X-ray diffraction and absorption, magnetometry, metrological surface measurements, highpressure diamond anvil cell equation-of-state, phonon dispersion measurements, X-ray absorption and emission spectroscopy, and differential scanning calorimetry. The sample preparation and materials processing capabilities in the IASPL have also facilitated experimentation at world-class facilities such as the

  12. Enforcement of the ban on aristolochic acids in Chinese traditional herbal preparations on the Dutch market.

    PubMed

    Martena, Martijn J; van der Wielen, Jacqueline C A; van de Laak, Leo F J; Konings, Erik J M; de Groot, Henk N; Rietjens, Ivonne M C M

    2007-09-01

    In traditional Chinese medicine several Aristolochia species are used. Aristolochia spp. contain a mixture of aristolochic acids (AAs), mainly AA I and AA II which are nephrotoxicants and carcinogens. After AA-related nephropathy (AAN) and urothelial cancer were described in female patients in Belgium following intake of AA-contaminated herbal preparations, herbs with AAs were prohibited worldwide. Confusing nomenclature can cause AA contamination of certain Chinese traditional herbal preparations (THPs). Here we report the results of investigations by the Dutch Food and Consumer Product Safety Authority (VWA) into the presence of AAs in THPs sampled on the Dutch market using a liquid-chromatography--mass spectrometry method. Between 2002 and 2006 we sampled 190 Chinese THPs using recent information on Chinese THPs potentially containing AAs. AA I was found in 25 samples up to a concentration of 1,676 mg/kg. AA II was also found in 13 of these samples up to 444 mg/kg. All 25 positive samples including Mu Tong, Fang Ji, Tian Xian Teng and Xi Xin were part of a group of 68 THPs identified as possibly containing AAs. In a worst-case scenario, use of a sample of Mu Tong with the highest AA content over a 7-day period would result in the same intake levels of AAs which significantly raised the cancer risk in the Belgian AAN cases. Our results show that contaminated THPs still can be found on the market following worldwide publicity. Therefore, it can be concluded that testing of possibly AA-contaminated THPs is still essential.

  13. Low-Cost 3D Printers Enable High-Quality and Automated Sample Preparation and Molecular Detection.

    PubMed

    Chan, Kamfai; Coen, Mauricio; Hardick, Justin; Gaydos, Charlotte A; Wong, Kah-Yat; Smith, Clayton; Wilson, Scott A; Vayugundla, Siva Praneeth; Wong, Season

    2016-01-01

    Most molecular diagnostic assays require upfront sample preparation steps to isolate the target's nucleic acids, followed by its amplification and detection using various nucleic acid amplification techniques. Because molecular diagnostic methods are generally rather difficult to perform manually without highly trained users, automated and integrated systems are highly desirable but too costly for use at point-of-care or low-resource settings. Here, we showcase the development of a low-cost and rapid nucleic acid isolation and amplification platform by modifying entry-level 3D printers that cost between $400 and $750. Our modifications consisted of replacing the extruder with a tip-comb attachment that houses magnets to conduct magnetic particle-based nucleic acid extraction. We then programmed the 3D printer to conduct motions that can perform high-quality extraction protocols. Up to 12 samples can be processed simultaneously in under 13 minutes and the efficiency of nucleic acid isolation matches well against gold-standard spin-column-based extraction technology. Additionally, we used the 3D printer's heated bed to supply heat to perform water bath-based polymerase chain reactions (PCRs). Using another attachment to hold PCR tubes, the 3D printer was programmed to automate the process of shuttling PCR tubes between water baths. By eliminating the temperature ramping needed in most commercial thermal cyclers, the run time of a 35-cycle PCR protocol was shortened by 33%. This article demonstrates that for applications in resource-limited settings, expensive nucleic acid extraction devices and thermal cyclers that are used in many central laboratories can be potentially replaced by a device modified from inexpensive entry-level 3D printers.

  14. Low-Cost 3D Printers Enable High-Quality and Automated Sample Preparation and Molecular Detection

    PubMed Central

    Chan, Kamfai; Coen, Mauricio; Hardick, Justin; Gaydos, Charlotte A.; Wong, Kah-Yat; Smith, Clayton; Wilson, Scott A.; Vayugundla, Siva Praneeth; Wong, Season

    2016-01-01

    Most molecular diagnostic assays require upfront sample preparation steps to isolate the target’s nucleic acids, followed by its amplification and detection using various nucleic acid amplification techniques. Because molecular diagnostic methods are generally rather difficult to perform manually without highly trained users, automated and integrated systems are highly desirable but too costly for use at point-of-care or low-resource settings. Here, we showcase the development of a low-cost and rapid nucleic acid isolation and amplification platform by modifying entry-level 3D printers that cost between $400 and $750. Our modifications consisted of replacing the extruder with a tip-comb attachment that houses magnets to conduct magnetic particle-based nucleic acid extraction. We then programmed the 3D printer to conduct motions that can perform high-quality extraction protocols. Up to 12 samples can be processed simultaneously in under 13 minutes and the efficiency of nucleic acid isolation matches well against gold-standard spin-column-based extraction technology. Additionally, we used the 3D printer’s heated bed to supply heat to perform water bath-based polymerase chain reactions (PCRs). Using another attachment to hold PCR tubes, the 3D printer was programmed to automate the process of shuttling PCR tubes between water baths. By eliminating the temperature ramping needed in most commercial thermal cyclers, the run time of a 35-cycle PCR protocol was shortened by 33%. This article demonstrates that for applications in resource-limited settings, expensive nucleic acid extraction devices and thermal cyclers that are used in many central laboratories can be potentially replaced by a device modified from inexpensive entry-level 3D printers. PMID:27362424

  15. Time of flight secondary ion mass spectrometry of bone-Impact of sample preparation and measurement conditions.

    PubMed

    Henss, Anja; Hild, Anne; Rohnke, Marcus; Wenisch, Sabine; Janek, Juergen

    2015-06-07

    Time of flight secondary ion mass spectrometry (ToF-SIMS) enables the simultaneous detection of organic and inorganic ions and fragments with high mass and spatial resolution. Due to recent technical developments, ToF-SIMS has been increasingly applied in the life sciences where sample preparation plays an eminent role for the quality of the analytical results. This paper focusses on sample preparation of bone tissue and its impact on ToF-SIMS analysis. The analysis of bone is important for the understanding of bone diseases and the development of replacement materials and new drugs for the cure of diseased bone. The main purpose of this paper is to find out which preparation process is best suited for ToF-SIMS analysis of bone tissue in order to obtain reliable and reproducible analytical results. The influence of the embedding process on the different components of bone is evaluated using principal component analysis. It is shown that epoxy resin as well as methacrylate based plastics (Epon and Technovit) as embedding materials do not infiltrate the mineralized tissue and that cut sections are better suited for the ToF-SIMS analysis than ground sections. In case of ground samples, a resin layer is smeared over the sample surface due to the polishing step and overlap of peaks is found. Beside some signals of fatty acids in the negative ion mode, the analysis of native, not embedded samples does not provide any advantage. The influence of bismuth bombardment and O2 flooding on the signal intensity of organic and inorganic fragments due to the variation of the ionization probability is additionally discussed. As C60 sputtering has to be applied to remove the smeared resin layer, its effect especially on the organic fragments of the bone is analyzed and described herein.

  16. Optimization of sample preparation for accurate results in quantitative NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Yamazaki, Taichi; Nakamura, Satoe; Saito, Takeshi

    2017-04-01

    Quantitative nuclear magnetic resonance (qNMR) spectroscopy has received high marks as an excellent measurement tool that does not require the same reference standard as the analyte. Measurement parameters have been discussed in detail and high-resolution balances have been used for sample preparation. However, the high-resolution balances, such as an ultra-microbalance, are not general-purpose analytical tools and many analysts may find those balances difficult to use, thereby hindering accurate sample preparation for qNMR measurement. In this study, we examined the relationship between the resolution of the balance and the amount of sample weighed during sample preparation. We were able to confirm the accuracy of the assay results for samples weighed on a high-resolution balance, such as the ultra-microbalance. Furthermore, when an appropriate tare and amount of sample was weighed on a given balance, accurate assay results were obtained with another high-resolution balance. Although this is a fundamental result, it offers important evidence that would enhance the versatility of the qNMR method.

  17. Electrodeposition as an alternate method for preparation of environmental samples for iodide by AMS

    DOE PAGES

    Adamic, M. L.; Lister, T. E.; Dufek, E. J.; ...

    2015-03-25

    This paper presents an evaluation of an alternate method for preparing environmental samples for 129I analysis by accelerator mass spectrometry (AMS) at Idaho National Laboratory. The optimal sample preparation method is characterized by ease of preparation, capability of processing very small quantities of iodide, and ease of loading into a cathode. Electrodeposition of iodide on a silver wire was evaluated using these criteria. This study indicates that the electrochemically-formed silver iodide deposits produce ion currents similar to those from precipitated silver iodide for the same sample mass. Furthermore, precipitated silver iodide samples are usually mixed with niobium or silver powdermore » prior to loading in a cathode. Using electrodeposition, the silver is already mixed with the sample and can simply be picked up with tweezers, placed in the sample die, and pressed into a cathode. The major advantage of this method is that the silver wire/electrodeposited silver iodide is much easier to load into a cathode.« less

  18. Field enhancement sample stacking for analysis of organic acids in traditional Chinese medicine by capillary electrophoresis.

    PubMed

    Zhu, Qianqian; Xu, Xueqin; Huang, Yuanyuan; Xu, Liangjun; Chen, Guonan

    2012-07-13

    A technique known as field enhancement sample stacking (FESS) and capillary electrophoresis (CE) separation has been developed to analyze and detect organic acids in the three traditional Chinese medicines (such as Portulaca oleracea L., Crataegus pinnatifida and Aloe vera L.). In FESS, a reverse electrode polarity-stacking mode (REPSM) was applied as on-line preconcentration strategy. Under the optimized condition, the baseline separation of eight organic acids (linolenic acid, lauric acid, p-coumaric acid, ascorbic acid, benzoic acid, caffeic acid, succinic acid and fumaric acid) could be achieved within 20 min. Validation parameters of this method (such as detection limits, linearity and precision) were also evaluated. The detection limits ranged from 0.4 to 60 ng/mL. The results indicated that the proposed method was effective for the separation of mixtures of organic acids. Satisfactory recoveries were also obtained in the analysis of these organic acids in the above traditional Chinese medicine samples.

  19. The influence of target preparation and mode of irradiation on PIXE analysis of biological samples

    NASA Astrophysics Data System (ADS)

    Galuszka, Janusz; Jarczyk, Lucjan; Rokita, Eugeniusz; Strzalkowski, Adam; Sych, Marek

    1984-04-01

    The following methods of target preparation were examined and compared: dry ashing at high temperature, low temperature ashing in plasma asher, wet ashing, lyophilization at a temperature of 35°C, cryofixation with drying in vacuum and dehydration in alcohol with drying in vacuum. All these techniques were applied to prepare targets from five different rat organs: liver, kidney, brain, lung and muscle tissue. The dried and powdered sample material was pressed into pellets or was distributed on formvar film. The evaporation of the thin carbon layer on the investigated target and placing of the thin carbon film in front of a target were also tested. The targets were irradiated in vacuum using an external beam in the air chamber. The influence of the method of target preparation on the detection limits, time requirements and escape of elements from the sample material is discussed.

  20. Optimized Ar(+)-ion milling procedure for TEM cross-section sample preparation.

    PubMed

    Dieterle, Levin; Butz, Benjamin; Müller, Erich

    2011-11-01

    High-quality samples are indispensable for every reliable transmission electron microscopy (TEM) investigation. In order to predict optimized parameters for the final Ar(+)-ion milling preparation step, topographical changes of symmetrical cross-section samples by the sputtering process were modeled by two-dimensional Monte-Carlo simulations. Due to its well-known sputtering yield of Ar(+)-ions and its easiness in mechanical preparation Si was used as model system. The simulations are based on a modified parameterized description of the sputtering yield of Ar(+)-ions on Si summarized from literature. The formation of a wedge-shaped profile, as commonly observed during double-sector ion milling of cross-section samples, was reproduced by the simulations, independent of the sputtering angle. Moreover, the preparation of wide, plane parallel sample areas by alternating single-sector ion milling is predicted by the simulations. These findings were validated by a systematic ion-milling study (single-sector vs. double-sector milling at various sputtering angles) using Si cross-section samples as well as two other material-science examples. The presented systematic single-sector ion-milling procedure is applicable for most Ar(+)-ion mills, which allow simultaneous milling from both sides of a TEM sample (top and bottom) in an azimuthally restricted sector perpendicular to the central epoxy line of that cross-sectional TEM sample. The procedure is based on the alternating milling of the two halves of the TEM sample instead of double-sector milling of the whole sample. Furthermore, various other practical aspects are issued like the dependency of the topographical quality of the final sample on parameters like epoxy thickness and incident angle.

  1. Thermophysical properties of starch and whey protein composite prepared in presence of organic acid and esters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previously, we prepared starch and protein composite by reactive mixing in presence of various organic acids and found that use of these acid esters resulted in composites with good mechanical properties. In this study, concentration (% w/w) of acid citrates in the starch-protein composites were var...

  2. Preparation of fatty acid methyl esters from Osage orange (Maclura pomifera) oil and evaluation as biodiesel

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fatty acid methyl esters were prepared in high yield by transesterification of Osage orange (Maclura pomifera) oil. Extracted using supercritical CO2, the crude oil was initially treated with mineral acid and methanol to lower its content of free fatty acids, thus rendering it amenable to homogeneou...

  3. 40 CFR 205.160-2 - Test sample selection and preparation.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Test sample selection and preparation. 205.160-2 Section 205.160-2 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) NOISE ABATEMENT PROGRAMS TRANSPORTATION EQUIPMENT NOISE EMISSION CONTROLS Motorcycles § 205.160-2...

  4. Soil and Water – What is Detectable through Microbiological Sample Preparation Techniques

    EPA Science Inventory

    The concerns of a potential terrorist’s use of biological agents in soil and ground water are articulated by comparisons to major illnesses in this Country involving contaminated drinking water sources. Objectives are focused on the importance of sample preparation in the rapid, ...

  5. Demonstration of a Sample Preparation Method for Biological Detection Based on a Novel Membrane Fractionation Technology

    DTIC Science & Technology

    2008-12-31

    oftarget nucleic acid in the sample) Deoxyribonucleic acid kilo Dalton Leonard Wood Institute Midwest Research Institute Quantitative Polymerase...membranes. Desalination 227: 111-119. 7) Kong S, Titchener- Hooker N, Levy MS, 2006. Plasmid DNA processing for gene therapy and vaccination: Studies on...Computerized Helical Scanning Technique. JAm Soc Nephrol, 13: S53-S61. 15. Bricefio, M.I. , Joseph , D.D. 2003. Self-lubricated transport of aqueous

  6. Stereochemistry of amino acids in surface samples of a marine sediment

    USGS Publications Warehouse

    Pollock, G.E.; Kvenvolden, K.A.

    1978-01-01

    In two surface samples of marine sediment, the percentages of d-alanine and d-aspartic acid are significantly higher than the other d-amino acids and are similar to the range found in soils. The percentage of d-glutamic acid is also higher than the other amino acids but less than d-alanine and d-aspartic acid. These d-amino acids may come mainly from bacteria. ?? 1978.

  7. Stereochemistry of amino acids in surface samples of a marine sediment

    NASA Technical Reports Server (NTRS)

    Pollock, G. E.; Kvenvolden, K. A.

    1978-01-01

    In two surface samples of marine sediment, the percentages of D-alanine and D-aspartic acid are significantly higher than the other D-amino acids and are similar to the range found in soils. The percentage of D-glutamic acid is also higher than the other amino acids but less than D-alanine and D-aspartic acid. These D-amino acids may come mainly from bacteria.

  8. Gas-assisted annular microsprayer for sample preparation for time-resolved cryo-electron microscopy

    NASA Astrophysics Data System (ADS)

    Lu, Zonghuan; Barnard, David; Shaikh, Tanvir R.; Meng, Xing; Mannella, Carmen A.; Yassin, Aymen S.; Agrawal, Rajendra K.; Wagenknecht, Terence; Lu, Toh-Ming

    2014-11-01

    Time-resolved cryo-electron microscopy (TRCEM) has emerged as a powerful technique for transient structural characterization of isolated biomacromolecular complexes in their native state within the time scale of seconds to milliseconds. For TRCEM sample preparation, a microfluidic device has been demonstrated to be a promising approach to facilitate TRCEM biological sample preparation. It is capable of achieving rapidly aqueous sample mixing, controlled reaction incubation, and sample deposition on electron microscopy (EM) grids for rapid freezing. One of the critical challenges is to transfer samples to cryo-EM grids from the microfluidic device. By using a microspraying method, the generated droplet size needs to be controlled to facilitate thin ice film formation on the grid surface for efficient data collection, whilst not being so thin that it dries out before freezing, i.e. an optimized mean droplet size needs to be achieved. In this work, we developed a novel monolithic three dimensional (3D) annular gas-assisted microfluidic sprayer using 3D MEMS (MicroElectroMechanical System) fabrication techniques. The microsprayer demonstrated dense and consistent microsprays with average droplet size between 6 and 9 μm, which fulfilled the droplet size requirement for TRCEM sample preparation. With droplet density of around 12-18 per grid window (window size 58  ×  58 μm), and a data collectible thin ice region of >50% total wetted area, we collected ~800-1000 high quality CCD micrographs in a 6-8 h period of continuous effort. This level of output is comparable to what were routinely achieving using cryo-grids prepared by conventional blotting and manual data collection. In this case, weeks of data collection with the previous device has been shortened to a day or two. And hundreds of microliters of valuable sample consumption can be reduced to only a small fraction.

  9. Gas-Assisted Annular Microsprayer for Sample Preparation for Time-Resolved Cryo-Electron Microscopy

    PubMed Central

    Lu, Zonghuan; Barnard, David; Shaikh, Tanvir R.; Meng, Xing; Mannella, Carmen A.; Yassin, Aymen; Agrawal, Rajendra; Wagenknecht, Terence; Lu, Toh-Ming

    2014-01-01

    Time-resolved cryo electron microscopy (TRCEM) has emerged as a powerful technique for transient structural characterization of isolated biomacromolecular complexes in their native state within the time scale of seconds to milliseconds. For TRCEM sample preparation, microfluidic device [9] has been demonstrated to be a promising approach to facilitate TRCEM biological sample preparation. It is capable of achieving rapidly aqueous sample mixing, controlled reaction incubation, and sample deposition on electron microscopy (EM) grids for rapid freezing. One of the critical challenges is to transfer samples to cryo-EM grids from the microfluidic device. By using microspraying method, the generated droplet size needs to be controlled to facilitate the thin ice film formation on the grid surface for efficient data collection, while not too thin to be dried out before freezing, i.e., optimized mean droplet size needs to be achieved. In this work, we developed a novel monolithic three dimensional (3D) annular gas-assisted microfluidic sprayer using 3D MEMS (MicroElectroMechanical System) fabrication techniques. The microsprayer demonstrated dense and consistent microsprays with average droplet size between 6-9 μm, which fulfilled the above droplet size requirement for TRCEM sample preparation. With droplet density of around 12-18 per grid window (window size is 58×58 μm), and the data collectible thin ice region of >50% total wetted area, we collected ~800-1000 high quality CCD micrographs in a 6-8 hour period of continuous effort. This level of output is comparable to what were routinely achieved using cryo-grids prepared by conventional blotting and manual data collection. In this case, weeks of data collection process with the previous device [9] has shortened to a day or two. And hundreds of microliter of valuable sample consumption can be reduced to only a small fraction. PMID:25530679

  10. Advancement of Solidification Processing Technology Through Real Time X-Ray Transmission Microscopy: Sample Preparation

    NASA Technical Reports Server (NTRS)

    Stefanescu, D. M.; Curreri, P. A.

    1996-01-01

    Two types of samples were prepared for the real time X-ray transmission microscopy (XTM) characterization. In the first series directional solidification experiments were carried out to evaluate the critical velocity of engulfment of zirconia particles in the Al and Al-Ni eutectic matrix under ground (l-g) conditions. The particle distribution in the samples was recorded on video before and after the samples were directionally solidified. In the second series samples of the above two type of composites were prepared for directional solidification runs to be carried out on the Advanced Gradient Heating Facility (AGHF) aboard the space shuttle during the LMS mission in June 1996. X-ray microscopy proved to be an invaluable tool for characterizing the particle distribution in the metal matrix samples. This kind of analysis helped in determining accurately the critical velocity of engulfment of ceramic particles by the melt interface in the opaque metal matrix composites. The quality of the cast samples with respect to porosity and instrumented thermocouple sheath breakage or shift could be easily viewed and thus helped in selecting samples for the space shuttle experiments. Summarizing the merits of this technique it can be stated that this technique enabled the use of cast metal matrix composite samples since the particle location was known prior to the experiment.

  11. Challenges of biological sample preparation for SIMS imaging of elements and molecules at subcellular resolution

    NASA Astrophysics Data System (ADS)

    Chandra, Subhash

    2008-12-01

    Secondary ion mass spectrometry (SIMS) based imaging techniques capable of subcellular resolution characterization of elements and molecules are becoming valuable tools in many areas of biology and medicine. Due to high vacuum requirements of SIMS, the live cells cannot be analyzed directly in the instrument. The sample preparation, therefore, plays a critical role in preserving the native chemical composition for SIMS analysis. This work focuses on the evaluation of frozen-hydrated and frozen freeze-dried sample preparations for SIMS studies of cultured cells with a CAMECA IMS-3f dynamic SIMS ion microscope instrument capable of producing SIMS images with a spatial resolution of 500 nm. The sandwich freeze-fracture method was used for fracturing the cells. The complimentary fracture planes in the plasma membrane were characterized by field-emission secondary electron microscopy (FESEM) in the frozen-hydrated state. The cells fractured at the dorsal surface were used for SIMS analysis. The frozen-hydrated SIMS analysis of individual cells under dynamic primary ion beam (O 2+) revealed local secondary ion signal enhancements correlated with the water image signals of 19(H 3O) +. A preferential removal of water from the frozen cell matrix in the Z-axis was also observed. These complications render the frozen-hydrated sample type less desirable for subcellular dynamic SIMS studies. The freeze-drying of frozen-hydrated cells, either inside the instrument or externally in a freeze-drier, allowed SIMS imaging of subcellular chemical composition. Morphological evaluations of fractured freeze-dried cells with SEM and confocal laser scanning microscopy (CLSM) revealed well-preserved mitochondria, Golgi apparatus, and stress fibers. SIMS analysis of fractured freeze-dried cells revealed well-preserved chemical composition of even the most highly diffusible ions like K + and Na + in physiologically relevant concentrations. The high K-low Na signature in individual cells

  12. Validation of Internal Controls for Extraction and Amplification of Nucleic Acids from Enteric Viruses in Water Samples ▿ †

    PubMed Central

    Hata, Akihiko; Katayama, Hiroyuki; Kitajima, Masaaki; Visvanathan, Chettiyappan; Nol, Chea; Furumai, Hiroaki

    2011-01-01

    Inhibitors that reduce viral nucleic acid extraction efficiency and interfere with cDNA synthesis and/or polymerase activity affect the molecular detection of viruses in aquatic environments. To overcome these significant problems, we developed a methodology for assessing nucleic acid yields and DNA amplification efficiencies for environmental water samples. This involved adding particles of adenovirus type 5 and murine norovirus and newly developed primer-sharing controls, which are amplified with the same primer pairs and result in the same amplicon sizes as the targets, to these samples. We found that nucleic acid loss during the extraction process, rather than reverse transcription-PCR (RT-PCR) inhibition, more significantly attributed to underestimation of the presence of viral genomes in the environmental water samples tested in this study. Our success rate for satisfactorily amplifying viral RNAs and DNAs by RT-PCR was higher than that for obtaining adequate nucleic acid preparations. We found that inhibitory properties were greatest when we used larger sample volumes. A magnetic silica bead-based RNA extraction method effectively removed inhibitors that interfere with viral nucleic acid extraction and RT-PCR. To our knowledge, this is the first study to assess the inhibitory properties of environmental water samples by using both control virus particles and primer-sharing controls. PMID:21602369

  13. Community-Level Physiological Profiling of Microbial Communities in Constructed Wetlands: Effects of Sample Preparation.

    PubMed

    Button, Mark; Weber, Kela; Nivala, Jaime; Aubron, Thomas; Müller, Roland Arno

    2016-03-01

    Community-level physiological profiling (CLPP) using BIOLOG® EcoPlates™ has become a popular method for characterizing and comparing the functional diversity, functional potential, and metabolic activity of heterotrophic microbial communities. The method was originally developed for profiling soil communities; however, its usage has expanded into the fields of ecotoxicology, agronomy, and the monitoring and profiling of microbial communities in various wastewater treatment systems, including constructed wetlands for water pollution control. When performing CLPP on aqueous samples from constructed wetlands, a wide variety of sample characteristics can be encountered and challenges may arise due to excessive solids, color, or turbidity. The aim of this study was to investigate the impacts of different sample preparation methods on CLPP performed on a variety of aqueous samples covering a broad range of physical and chemical characteristics. The results show that using filter paper, centrifugation, or settling helped clarify samples for subsequent CLPP analysis, however did not do so as effectively as dilution for the darkest samples. Dilution was able to provide suitable clarity for the darkest samples; however, 100-fold dilution significantly affected the carbon source utilization patterns (CSUPs), particularly with samples that were already partially or fully clear. Ten-fold dilution also had some effect on the CSUPs of samples which were originally clear; however, the effect was minimal. Based on these findings, for this specific set of samples, a 10-fold dilution provided a good balance between ease of use, sufficient clarity (for dark samples), and limited effect on CSUPs. The process and findings outlined here can hopefully serve future studies looking to utilize CLPP for functional analysis of microbial communities and also assist in comparing data from studies where different sample preparation methods were utilized.

  14. Applied Focused Ion Beam Techniques for Sample Preparation of Astromaterials for Integrated Nano-Analysis

    SciTech Connect

    Graham, G A; Teslich, N E; Kearsley, A T; Stadermann, F J; Stroud, R M; Dai, Z R; Ishii, H A; Hutcheon, I D; Bajt, S; Snead, C J; Weber, P K; Bradley, J P

    2007-02-20

    Sample preparation is always a critical step in study of micrometer sized astromaterials available for study in the laboratory, whether their subsequent analysis is by electron microscopy or secondary ion mass spectrometry. A focused beam of gallium ions has been used to prepare electron transparent sections from an interplanetary dust particle, as part of an integrated analysis protocol to maximize the mineralogical, elemental, isotopic and spectroscopic information extracted from one individual particle. In addition, focused ion beam techniques have been employed to extract cometary residue preserved on the rims and walls of micro-craters in 1100 series aluminum foils that were wrapped around the sample tray assembly on the Stardust cometary sample collector. Non-ideal surface geometries and inconveniently located regions of interest required creative solutions. These include support pillar construction and relocation of a significant portion of sample to access a region of interest. Serial sectioning, in a manner similar to ultramicrotomy, is a significant development and further demonstrates the unique capabilities of focused ion beam microscopy for sample preparation of astromaterials.

  15. Yeast metabolomics: sample preparation for a GC/MS-based analysis.

    PubMed

    Carneiro, Sónia; Pereira, Rui; Rocha, Isabel

    2014-01-01

    Metabolome sample preparation is one of the key factors in metabolomics analyses. The quality of the metabolome data will depend on the suitability of the experimental procedures to the cellular system (e.g., yeast cells) and the analytical performance. Here, we summarize a protocol for metabolome analysis of yeast cells using gas chromatography-mass spectrometry (GC-MS). First, the main phases of a metabolomics analysis are identified: sample preparation, metabolite extraction, and analysis. We also provide an overview on different methods used to quench samples and extract intracellular metabolites from yeast cells. This protocol provides a detailed description of a GC-MS-based analysis of yeast metabolome, in particular for metabolites containing amino and/or carboxyl groups, which represent most of the compounds participating in the central carbon metabolism.

  16. Sample preparation of organic liquid for off-site analysis of chemical weapons convention related compounds.

    PubMed

    Pardasani, Deepak; Palit, Meehir; Gupta, A K; Shakya, Purushottam; Sekhar, K; Dubey, D K

    2005-02-15

    Off-site analysis of chemical warfare agents (CWAs) and related compounds plays a key role in the verification program of the Chemical Weapons Convention (CWC). The analysis results, aiming toward unambiguous identication of compounds, depend on the type of sample preparation method. Development of milder sample preparation methods, which offer good recoveries and do not alter the structure of analytes, is highly desirable. Organic liquid with high hydrocarbon background is a frequently encountered challenge in off-site analysis and in official proficiency tests conducted by OPCW. Sample cleanup procedures, namely, solvent exchange followed by cooling and liquid-liquid extraction were studied to eliminate the hydrocarbons from organic liquid. Acetonitrile, a polar aprotic solvent, was effectively used to remove the background in both methods, and recoveries of spiked CWAs by the two techniques were between 69 and 99%.

  17. The planning and establishment of a sample preparation laboratory for drug discovery

    PubMed Central

    Dufresne, Claude

    2000-01-01

    Nature has always been a productive source of new drugs. With the advent of high-throughput screening, it has now become possible to rapidly screen large sample collections. In addition to seeking greater diversity from natural product sources (micro-organisms, plants, etc.), fractionation of the crude extracts prior to screening is becoming a more important part of our efforts. As sample preparation protocols become more involved, automation can help to achieve and maintain a desired sample throughput. To address the needs of our screening program, two robotic systems were designed. The first system processes crude extracts all the way to 96-well plates, containing solutions suitable for screening in biological and biochemical assays. The system can dissolve crude extracts, fractionate them on solid-phase extraction cartridges, dry and weigh each fraction, re-dissolve them to a known concentration, and prepare mother plates. The second system replicates mother plates into a number of daughter plates. PMID:18924691

  18. Determination of deoxycholic acid pool size and input rate using (24-/sup 13/C)deoxycholic acid and serum sampling

    SciTech Connect

    Stellard, F.; Paumgartner, G.; van Berge Henegouwen, G.P.; van der Werf, S.D.

    1986-11-01

    We have developed an isotope dilution method for determination of deoxycholic acid pool size and input rate which employs oral administration of 50 mg of (24-/sup 13/C)deoxycholic acid and serum sampling. The method has been validated by classical isotope dilution technique using (24-/sup 14/C)deoxycholic acid and bile sampling in five patients with colonic adenomas. Excellent agreement between pool sizes and input rates determined with /sup 13/C/12C isotope ratio measurements in serum and /sup 14/C measurements in bile was obtained when isotope ratios were measured in the conjugated fraction of deoxycholic acid in serum. We conclude that pool size and input rate of deoxycholic acid can accurately be determined by blood sampling after oral administration of (24-/sup 13/C)deoxycholic acid, therewith eliminating the use of radioactive tracers and the need for bile sampling.

  19. Isolation and purification of heroin from heroin street samples by preparative high performance liquid chromatography.

    PubMed

    Guo, Zhen; Zheng, Hui; Lu, Yanzhen; Wei, Yun

    2012-09-10

    The present study established a novel method using preparative high performance liquid chromatography to isolate and purify heroin·HCl from heroin street samples to be used as a reference standard. Different kinds of mobile phases and columns were used, ultimately the mobile phase consisting of hexane-isopropanol-methanol (65:28:7, v/v) and the SIL preparative column prepared in laboratory were selected as the final condition. Heroin was further purified by the drowning-out crystallization method using isopropanol-methanol (50:1, v/v) and hexane as drowning-out anti-solvents and salting-out agents, respectively. The purity was assessed by analytical high performance liquid chromatography and the confirmation of the chemical structure was performed by IR and NMR. About 110.7mg of heroin·HCl at a purity of over 99.52% was obtained from 180mg of heroin street samples which contained 156.15mg of heroin·HCl component by preparative high performance liquid chromatography. This method is suitable for preparing heroin standards in forensic science area.

  20. An enzyme-based DNA preparation method for application to forensic biological samples and degraded stains.

    PubMed

    Lounsbury, Jenny A; Coult, Natalie; Miranian, Daniel C; Cronk, Stephen M; Haverstick, Doris M; Kinnon, Paul; Saul, David J; Landers, James P

    2012-09-01

    Extraction of DNA from forensic samples typically uses either an organic extraction protocol or solid phase extraction (SPE) and these methods generally involve numerous sample transfer, wash and centrifugation steps. Although SPE has been successfully adapted to the microdevice, it can be problematic because of lengthy load times and uneven packing of the solid phase. A closed-tube enzyme-based DNA preparation method has recently been developed which uses a neutral proteinase to lyse cells and degrade proteins and nucleases [14]. Following a 20 min incubation of the buccal or whole blood sample with this proteinase, DNA is polymerase chain reaction (PCR)-ready. This paper describes the optimization and quantitation of DNA yield using this method, and application to forensic biological samples, including UV- and heat-degraded whole blood samples on cotton or blue denim substrates. Results demonstrate that DNA yield can be increased from 1.42 (±0.21)ng/μL to 7.78 (±1.40)ng/μL by increasing the quantity of enzyme per reaction by 3-fold. Additionally, there is a linear relationship between the amount of starting cellular material added and the concentration of DNA in the solution, thereby allowing DNA yield estimations to be made. In addition, short tandem repeat (STR) profile results obtained using DNA prepared with the enzyme method were comparable to those obtained with a conventional SPE method, resulting in full STR profiles (16 of 16 loci) from liquid samples (buccal swab eluate and whole blood), dried buccal swabs and bloodstains and partial profiles from UV or heat-degraded bloodstains on cotton or blue denim substrates. Finally, the DNA preparation method is shown to be adaptable to glass or poly(methyl methacrylate) (PMMA) microdevices with little impact on STR peak height but providing a 20-fold reduction in incubation time (as little as 60 s), leading to a ≥1 h reduction in DNA preparation time.

  1. Amino acid preservation in saline halite core samples: Analogs for Martian dry evaporitic regions

    NASA Astrophysics Data System (ADS)

    Bada, J.; Aubrey, A.; Lowenstein, T.; Timofeeff, M.

    2008-12-01

    in the deepest core section. This may indicate some recent amino acid contribution to the pool of certain amino acids. Racemization rates can be calculated from the equation: ln[(1+D/L)/(1-D/L)] - ln [(1+D/L)/(1-D/L)]t=0 = 2ki(time) where ki is the first-order rate constant for the interconversion of the enantiomers. Using the D/L ratios at the top of the core for the t = 0 term gives kasp = 3.5x10exp-5 y-1 and 1.3x10exp-5 y-1 for the 18 and 70 ka samples, respectively. For valine, the values are kval = 5.6x10exp-6 y-1 and 7.3x10exp-6 y-1. Extrapolating these values to the average surface temperatures on Mars indicates that the chirality of these amino acids would be preserved for billions of years. Thus, closed basin lacustrine and dry desert valley regions with evaporite-rich deposits are suitable environments in the search for preserved biosignatures on Mars. References [1] Bibring, J.P., et al., Science 307, 1576 (2005) [2] Klinghofer, G., et al., Science 306, 1740 (2004) [3] Osterloo, M.M., et al., Science 319, 1651 (2008) [4] Squyres, S.W., et al., Nature 443, E1 (2006) [5] Lowenstein, T.K., et al., Geology 27, 3 (1999) [6] Glavin, D., et al., Earth Planet. Sci. Lett. 185,1 (2001) [7] Aubrey, A. D., et al., in preparation, Nature Geo. Sci.

  2. Novel Sample Preparation Technique To Improve Spectromicroscopic Analyses of Micrometer-Sized Particles.

    PubMed

    Höschen, Carmen; Höschen, Till; Mueller, Carsten W; Lugmeier, Johann; Elgeti, Stefan; Rennert, Thilo; Kögel-Knabner, Ingrid

    2015-08-18

    Microscale processes occurring at biogeochemical interfaces in soils and sediments have fundamental impacts on phenomena at larger scales. To obtain the organo-mineral associations necessary for the study of biogeochemical interfaces, bulk samples are usually fractionated into microaggregates or micrometer-sized single particles. Such fine-grained mineral particles are often prepared for nanoscale secondary ion mass spectroscopy (NanoSIMS) investigations by depositing them on a carrier. This introduces topographic differences, which can strongly affect local sputtering efficiencies. Embedding in resin causes undesired C impurities. We present a novel method for preparing polished cross-sections of micrometer-sized primary soil particles that overcomes the problems of topography and C contamination. The particles are coated with a marker layer, embedded, and well-polished. The interpretation of NanoSIMS data is assisted by energy-dispersive X-ray spectroscopy on cross-sections prepared by a focused ion beam. In the cross-sections, organic assemblages on the primary soil particles become visible. This novel method significantly improves the quality of NanoSIMS measurements on grainy mineral samples, enabling better characterization of soil biogeochemical interfaces. In addition, this sample preparation technique may also improve results from other (spectro-) microscopic techniques.

  3. Guidance document for preparing water sampling and analysis plans for UMTRA sites. Final [report

    SciTech Connect

    Not Available

    1993-09-01

    A water sampling and analysis plan (WSAP) is required for each Uranium Mill Tailings Remedial Action (UMTRA) site to provide rationale for groundwater and surface water sampling at disposal sites and former processing sites. The WSAP identifies and justifies the sampling locations, analytical parameters, detection limits, and sampling frequencies for the groundwater and surface water monitoring stations at each site. Section 2.0 of this WSAP Guidance Document describes the WSAP format. Sections 3.0 and 4.0 provide guidance for selecting sampling frequencies and sampling locations, respectively. Section 5.0 contains criteria for selecting analytical parameters. Section 6.0 provides guidance for the contents of each site`s WSAP file. Finally, Section 7.0 presents the references used to prepare this document. The purpose of this guidance document is to provide a consistent technical approach for sampling and monitoring activities performed under WSAPs and a consistent format for WSAP documents. This document is designed for use by the Technical Assistance Contractor (TAC) to prepare WSAPs and by the US Department of Energy (DOE), Nuclear Regulatory Commission, state and tribal agencies, regulatory agencies, and the public to evaluate the contents of the WSAPS. This guidance document may be updated periodically based on new or changing regulations.

  4. On the abiotic formation of amino acids. I - HCN as a precursor of amino acids detected in extracts of lunar samples. II - Formation of HCN and amino acids from simulated mixtures of gases released from lunar samples

    NASA Technical Reports Server (NTRS)

    Yuasa, S.; Flory, D.; Basile, B.; Oro, J.

    1984-01-01

    Two studies on the abiotic formation of amino acids are presented. The first study demonstrates the role of hydrogen cyanide as a precursor of amino acids detected in extracts of lunar samples. The formation of several amino acids, including glycine, alanine, aspartic acid, and glutamic acid, under conditions similar to those used for the analysis of lunar samples is demonstrated. The second study investigates the formation of hydrogen cyanide as well as amino acids from lunar-sample gas mixtures under electrical discharge conditions. These results extend the possibility of synthesis of amino acids to planetary bodies with primordial atmospheres less reducing than a mixture of methane, ammonia, hydrogen and water.

  5. SAMPLING ARTIFACT ESTIMATES FOR ALKANES, HOPANES, AND ALIPHATIC CARBOXYLIC ACIDS

    EPA Science Inventory

    Sampling artifacts for molecular markers from organic speciation of particulate matter were investigated by analyzing forty-one samples collected in Philadelphia as a part of the Northeast Oxidant and Particulate Study (NEOPS). Samples were collected using a high volume sampler ...

  6. Sequential injection titration method using second-order signals: determination of acidity in plant oils and biodiesel samples.

    PubMed

    del Río, Vanessa; Larrechi, M Soledad; Callao, M Pilar

    2010-06-15

    A new concept of flow titration is proposed and demonstrated for the determination of total acidity in plant oils and biodiesel. We use sequential injection analysis (SIA) with a diode array spectrophotometric detector linked to chemometric tools such as multivariate curve resolution-alternating least squares (MCR-ALS). This system is based on the evolution of the basic specie of an acid-base indicator, alizarine, when it comes into contact with a sample that contains free fatty acids. The gradual pH change in the reactor coil due to diffusion and reaction phenomenona allows the sequential appearance of both species of the indicator in the detector coil, recording a data matrix for each sample. The SIA-MCR-ALS method helps to reduce the amounts of sample, the reagents and the time consumed. Each determination consumes 0.413ml of sample, 0.250ml of indicator and 3ml of carrier (ethanol) and generates 3.333ml of waste. The frequency of the analysis is high (12 samples h(-1) including all steps, i.e., cleaning, preparing and analysing). The utilized reagents are of common use in the laboratory and it is not necessary to use the reagents of perfect known concentration. The method was applied to determine acidity in plant oil and biodiesel samples. Results obtained by the proposed method compare well with those obtained by the official European Community method that is time consuming and uses large amounts of organic solvents.

  7. Use of nitric acid in sample pretreatment for determination of trace elements in various biological samples by ETAAS.

    PubMed

    Scancar, J; Milacic, R; Falnoga, I; Cemazar, M; Bukovec, P

    2000-07-01

    Trace elements in liquid biological samples may be determined by direct electrothermal atomic absorption spectrometry (ETAAS). In our previous work it was found that samples containing proteins or DNA may leak out of the graphite tube before the drying step, despite the addition of various modifiers. In order to keep the sample to the graphite tube, samples were diluted before analysis 1 + 1 with 32% v/v nitric acid, or 5 microl of 32% v/v nitric acid was added to the graphite tube before ETAAS determination. Applying the proposed procedure, the concentrations of lead in eluted fractions after gel chromatographic separation of human cerebellar nucleus dentatus supernatant and platinum in isolated DNA samples were determined. The use of nitric acid in sample pretreatment prevent sample leakage out of the graphite tube, provided for even drying and considerably reduced nonspecific absorption in lead determination. The repeatability of measurements was better than + 6%. The accuracy of the procedure was checked by spiking samples. The recoveries for both elements lay between 93--104%. Nitric acid was found to be a better modifier than TRITON X-100.

  8. Preparation and tribological properties of stearic acid-modified hierarchical anatase TiO 2 microcrystals

    NASA Astrophysics Data System (ADS)

    Qian, Jianhua; Yin, Xiangyu; Wang, Ning; Liu, Lin; Xing, Jinjuan

    2012-01-01

    Hierarchical TiO2 microcrystals were synthesized through a facile solvothermal method. X-ray diffraction (XRD) and scanning electron microscope (SEM) measurements were used to characterize the structure of the as-prepared samples. The results indicated that the synthesized hierarchical titania (TiO2) microspheres were composed of numerous anatase phase TiO2 particles. The as-prepared samples were chemically modified with stearic acid to improve their dispersion in oil. Fourier transmission infrared spectroscopy (FT-IR) and thermogravimetry analysis (TGA) were carried out to evaluate the characteristics of the modified TiO2 microcrystals. The tribological properties of the modified TiO2 microcrystals as additives of liquid paraffin were studied by a four-ball tester, and the results showed that they could significantly improve anti-wear performance, friction-reduction property and load-carrying capacity of liquid paraffin. These advantages make the modified TiO2 microcrystals promising for green lubricating oil additives.

  9. Graphite sample preparation for AMS in a high pressure and temperature press

    USGS Publications Warehouse

    Rubin, Meyer; Mysen, Bjorn O.; Polach, Henry

    1984-01-01

    A high pressure-temperature press is used to make target material for accelerator mass spectrometry. Graphite was produced from typical **1**4C samples including oxalic acid and carbonates. Beam strength of **1**2C was generally adequate, but random radioactive contamination by **1**4C made age measurements impractical.

  10. Graphite sample preparation for AMS in a high pressure and temperature press

    USGS Publications Warehouse

    Rubin, M.; Mysen, B.O.; Polach, H.

    1984-01-01

    A high pressure-high temperature press is used to make target material for accelerator mass spectrometry. Graphite was produced from typical 14C samples including oxalic acid and carbonates. Beam strength of 12C was generally adequate, but random radioactive contamination by 14C made age measurements impractical. ?? 1984.

  11. Cytology Preparations of Formalin Fixative Aid Detection of Giardia in Duodenal Biopsy Samples.

    PubMed

    Panarelli, Nicole C; Gobara, Nariman; Hoda, Rana S; Chaump, Michael; Jessurun, Jose; Yantiss, Rhonda K

    2017-04-01

    Giardiasis is the most common intestinal parasitic infection in the United States. The organism elicits no, or minimal, inflammatory changes in duodenal biopsy samples, so it can be easily overlooked. We performed this study to determine whether Giardia could be isolated from the formalin fixative of biopsy samples, and to evaluate the value of fluid analysis in the assessment for potential infection. We prospectively evaluated duodenal biopsy samples from 92 patients with a clinical suspicion of giardiasis or symptoms compatible with that diagnosis (ie, diarrhea, bloating, or abdominal pain) Biopsy samples were routinely processed and stained with hematoxylin and eosin. Histologic diagnoses included giardiasis (5 cases, 4%), normal findings (64 cases, 70%), peptic injury/active duodenitis (12 cases, 13%), and intraepithelial lymphocytosis with villous blunting (10 cases, 12%). Fifteen cases (13%) showed detached degenerated epithelial cells or mucus droplets in the intervillous space that resembled Giardia. Cytology slides were prepared from formalin in the biopsy container using the standard Cytospin protocol and reviewed by a cytopathologist blinded to the biopsy findings. Cytologic evaluation revealed Giardia spp. in all 5 biopsy-proven cases, and identified an additional case that was not detected by biopsy analysis. Organisms were significantly more numerous (mean: 400 trophozoites; range, 120 to 810) and showed better morphologic features in cytology preparations compared with tissue sections (mean: 129 trophozoites; range, 37 to 253 organisms; P=0.05). Our findings suggest that cytology preparations from formalin fixative can resolve diagnostically challenging cases and even enhance Giardia detection in some cases.

  12. Multiplexed efficient on-chip sample preparation and sensitive amplification-free detection of Ebola virus.

    PubMed

    Du, K; Cai, H; Park, M; Wall, T A; Stott, M A; Alfson, K J; Griffiths, A; Carrion, R; Patterson, J L; Hawkins, A R; Schmidt, H; Mathies, R A

    2017-05-15

    An automated microfluidic sample preparation multiplexer (SPM) has been developed and evaluated for Ebola virus detection. Metered air bubbles controlled by microvalves are used to improve bead-solution mixing thereby enhancing the hybridization of the target Ebola virus RNA with capture probes bound to the beads. The method uses thermally stable 4-formyl benzamide functionalized (4FB) magnetic beads rather than streptavidin coated beads with a high density of capture probes to improve the target capture efficiency. Exploiting an on-chip concentration protocol in the SPM and the single molecule detection capability of the antiresonant reflecting optical waveguide (ARROW) biosensor chip, a detection limit of 0.021pfu/mL for clinical samples is achieved without target amplification. This RNA target capture efficiency is two orders of magnitude higher than previous results using streptavidin beads and the limit of detection (LOD) improves 10×. The wide dynamic range of this technique covers the whole clinically applicable concentration range. In addition, the current sample preparation time is ~1h which is eight times faster than previous work. This multiplexed, miniaturized sample preparation microdevice establishes a key technology that intended to develop next generation point-of-care (POC) detection system.

  13. Molecularly imprinted polymers for sample preparation and biosensing in food analysis: Progress and perspectives.

    PubMed

    Ashley, Jon; Shahbazi, Mohammad-Ali; Kant, Krishna; Chidambara, Vinayaka Aaydha; Wolff, Anders; Bang, Dang Duong; Sun, Yi

    2017-05-15

    Molecularly imprinted polymers (MIPs) are biomimetics which can selectively bind to analytes of interest. One of the most interesting areas where MIPs have shown the biggest potential is food analysis. MIPs have found use as sorbents in sample preparation attributed to the high selectivity and high loading capacity. MIPs have been intensively employed in classical solid-phase extraction and solid-phase microextraction. More recently, MIPs have been combined with magnetic bead extraction, which greatly simplifies sample handling procedures. Studies have consistently shown that MIPs can effectively minimize complex food matrix effects, and improve recoveries and detection limits. In addition to sample preparation, MIPs have also been viewed as promising alternatives to bio-receptors due to the inherent molecular recognition abilities and the high stability in harsh chemical and physical conditions. MIPs have been utilized as receptors in biosensing platforms such as electrochemical, optical and mass biosensors to detect various analytes in food. In this review, we will discuss the current state-of-the-art of MIP synthesis and applications in the context of food analysis. We will highlight the imprinting methods which are applicable for imprinting food templates, summarize the recent progress in using MIPs for preparing and analysing food samples, and discuss the current limitations in the commercialisation of MIPs technology. Finally, future perspectives will be given.

  14. Improvements in preparation of samples generated by SW-846 Method 0010

    SciTech Connect

    Jackson, M.D.; Johnson, L.D.; McGaughey, J.F.; Wagoner, D.E.; Bursey, J.T.; Merrill, R.G.

    1994-12-31

    A field evaluation study for SW-846 Method 0010 was conducted at a stationary source emission site with a high moisture content. The recovery of the dynamically spiked analytes and the spikes added before the laboratory preparation of the XAD-2{reg_sign} resin were low. The sampling train media had been prepared according to Method 0010 protocol, so the laboratory procedures were examined in detail for sources of error. The XAD-2{reg_sign}, wet because of the source matrix, was difficult to fully remove from the glass trap using only methylene chloride. Because Method 0010 does not specify the solvent to use for trap rinsing, and since the probe was washed in the field with a 50:50 mixture of methylene chloride and methanol, this mixture was used to transfer the XAD-2{reg_sign} from the trap to the Soxhlet extractor. The low recoveries were attributed to the presence of the methanol in the final extract. (The methanol is removed from the probe rinses during the laboratory sample preparation process.) A proposed method will be presented that permits the physical removal of wet XAD-2{reg_sign} from the sampling trap without the use of methanol and an alternate procedure is provided for removal of methanol from the trap rinse before sample extract concentration.

  15. Compact low temperature scanning tunneling microscope with in-situ sample preparation capability

    NASA Astrophysics Data System (ADS)

    Kim, Jungdae; Nam, Hyoungdo; Qin, Shengyong; Kim, Sang-ui; Schroeder, Allan; Eom, Daejin; Shih, Chih-Kang

    2015-09-01

    We report on the design of a compact low temperature scanning tunneling microscope (STM) having in-situ sample preparation capability. The in-situ sample preparation chamber was designed to be compact allowing quick transfer of samples to the STM stage, which is ideal for preparing temperature sensitive samples such as ultra-thin metal films on semiconductor substrates. Conventional spring suspensions on the STM head often cause mechanical issues. To address this problem, we developed a simple vibration damper consisting of welded metal bellows and rubber pads. In addition, we developed a novel technique to ensure an ultra-high-vacuum (UHV) seal between the copper and stainless steel, which provides excellent reliability for cryostats operating in UHV. The performance of the STM was tested from 2 K to 77 K by using epitaxial thin Pb films on Si. Very high mechanical stability was achieved with clear atomic resolution even when using cryostats operating at 77 K. At 2 K, a clean superconducting gap was observed, and the spectrum was easily fit using the BCS density of states with negligible broadening.

  16. Compact low temperature scanning tunneling microscope with in-situ sample preparation capability

    SciTech Connect

    Kim, Jungdae; Nam, Hyoungdo; Schroeder, Allan; Shih, Chih-Kang; Qin, Shengyong; Kim, Sang-ui; Eom, Daejin

    2015-09-15

    We report on the design of a compact low temperature scanning tunneling microscope (STM) having in-situ sample preparation capability. The in-situ sample preparation chamber was designed to be compact allowing quick transfer of samples to the STM stage, which is ideal for preparing temperature sensitive samples such as ultra-thin metal films on semiconductor substrates. Conventional spring suspensions on the STM head often cause mechanical issues. To address this problem, we developed a simple vibration damper consisting of welded metal bellows and rubber pads. In addition, we developed a novel technique to ensure an ultra-high-vacuum (UHV) seal between the copper and stainless steel, which provides excellent reliability for cryostats operating in UHV. The performance of the STM was tested from 2 K to 77 K by using epitaxial thin Pb films on Si. Very high mechanical stability was achieved with clear atomic resolution even when using cryostats operating at 77 K. At 2 K, a clean superconducting gap was observed, and the spectrum was easily fit using the BCS density of states with negligible broadening.

  17. Compact low temperature scanning tunneling microscope with in-situ sample preparation capability.

    PubMed

    Kim, Jungdae; Nam, Hyoungdo; Qin, Shengyong; Kim, Sang-ui; Schroeder, Allan; Eom, Daejin; Shih, Chih-Kang

    2015-09-01

    We report on the design of a compact low temperature scanning tunneling microscope (STM) having in-situ sample preparation capability. The in-situ sample preparation chamber was designed to be compact allowing quick transfer of samples to the STM stage, which is ideal for preparing temperature sensitive samples such as ultra-thin metal films on semiconductor substrates. Conventional spring suspensions on the STM head often cause mechanical issues. To address this problem, we developed a simple vibration damper consisting of welded metal bellows and rubber pads. In addition, we developed a novel technique to ensure an ultra-high-vacuum (UHV) seal between the copper and stainless steel, which provides excellent reliability for cryostats operating in UHV. The performance of the STM was tested from 2 K to 77 K by using epitaxial thin Pb films on Si. Very high mechanical stability was achieved with clear atomic resolution even when using cryostats operating at 77 K. At 2 K, a clean superconducting gap was observed, and the spectrum was easily fit using the BCS density of states with negligible broadening.

  18. Review of sample preparation techniques for the analysis of pesticide residues in soil.

    PubMed

    Tadeo, José L; Pérez, Rosa Ana; Albero, Beatriz; García-Valcárcel, Ana I; Sánchez-Brunete, Consuelo

    2012-01-01

    This paper reviews the sample preparation techniques used for the analysis of pesticides in soil. The present status and recent advances made during the last 5 years in these methods are discussed. The analysis of pesticide residues in soil requires the extraction of analytes from this matrix, followed by a cleanup procedure, when necessary, prior to their instrumental determination. The optimization of sample preparation is a very important part of the method development that can reduce the analysis time, the amount of solvent, and the size of samples. This review considers all aspects of sample preparation, including extraction and cleanup. Classical extraction techniques, such as shaking, Soxhlet, and ultrasonic-assisted extraction, and modern techniques like pressurized liquid extraction, microwave-assisted extraction, solid-phase microextraction and QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) are reviewed. The different cleanup strategies applied for the purification of soil extracts are also discussed. In addition, the application of these techniques to environmental studies is considered.

  19. Homogeneous immunosubtraction integrated with sample preparation is enabled by a microfluidic format

    PubMed Central

    Apori, Akwasi A.; Herr, Amy E.

    2011-01-01

    Immunosubtraction is a powerful and resource-intensive laboratory medicine assay that reports both protein mobility and binding specificity. To expedite and automate this electrophoretic assay, we report on advances to the electrophoretic immunosubtraction assay by introducing a homogeneous, not heterogeneous, format with integrated sample preparation. To accomplish homogeneous immunosubtraction, a step-decrease in separation matrix pore-size at the head of a polyacrylamide gel electrophoresis (PAGE) separation channel enables ‘subtraction’ of target analyte when capture antibody is present (as the large immune-complex is excluded from PAGE), but no subtraction when capture antibody is absent. Inclusion of sample preparation functionality via small pore size polyacrylamide membranes is also key to automated operation (i.e., sample enrichment, fluorescence sample labeling, and mixing of sample with free capture antibody). Homogenous sample preparation and assay operation allows on-the-fly, integrated subtraction of one to multiple protein targets and reuse of each device. Optimization of the assay is detailed which allowed for ~95% subtraction of target with 20% non-specific extraction of large species at the optimal antibody-antigen ratio, providing conditions needed for selective target identification. We demonstrate the assay on putative markers of injury and inflammation in cerebrospinal fluid (CSF), an emerging area of diagnostics research, by rapidly reporting protein mobility and binding specificity within the sample matrix. We simultaneously detect S100B and C-reactive protein, suspected biomarkers for traumatic brain injury (TBI), in ~2 min. Lastly, we demonstrate S100B detection (65 nM) in raw human CSF with a lower limit of detection of ~3.25 nM, within the clinically relevant concentration range for detecting TBI in CSF. Beyond the novel CSF assay introduced here, a fully automated immunosubtraction assay would impact a spectrum of routine but labor

  20. Oxalic-acid leaching of rock, soil, and stream-sediment samples as an anomaly-accentuated technique

    USGS Publications Warehouse

    Alminas, Henry V.; Mosier, Elwin L.

    1976-01-01

    In many instances total-rock and sieved-soil and stream-sediment samples lack the sensitivity and contrast required for reconnaissance exploration and necessary in the search for blind ore deposits. Heavy-mineral concentrates incorporate the required sensitivity and contrast but are overly expensive for two reasons: time-consuming sample preparation is required to obtain them, and they cannot be easily derived from all bulk-sample types. Trace-metal-content comparisons of the oxalic-acid-leachable portions with heavy-mineral concentrates show that the leachates are equal to the heavy-mineral concentrates in sensitivity and contrast. Simplicity of preparation and the resultant cost savings are additional advantages of this proposed method.

  1. imFASP: An integrated approach combining in-situ filter-aided sample pretreatment with microwave-assisted protein digestion for fast and efficient proteome sample preparation.

    PubMed

    Zhao, Qun; Fang, Fei; Wu, Ci; Wu, Qi; Liang, Yu; Liang, Zhen; Zhang, Lihua; Zhang, Yukui

    2016-03-17

    An integrated sample preparation method, termed "imFASP", which combined in-situ filter-aided sample pretreatment and microwave-assisted trypsin digestion, was developed for preparation of microgram and even nanogram amounts of complex protein samples with high efficiency in 1 h. For imFASP method, proteins dissolved in 8 M urea were loaded onto a filter device with molecular weight cut off (MWCO) as 10 kDa, followed by in-situ protein preconcentration, denaturation, reduction, alkylation, and microwave-assisted tryptic digestion. Compared with traditional in-solution sample preparation method, imFASP method generated more protein and peptide identifications (IDs) from preparation of 45 μg Escherichia coli protein sample due to the higher efficiency, and the sample preparation throughput was significantly improved by 14 times (1 h vs. 15 h). More importantly, when the starting amounts of E. coli cell lysate decreased to nanogram level (50-500 ng), the protein and peptide identified by imFASP method were improved at least 30% and 44%, compared with traditional in-solution preparation method, suggesting dramatically higher peptide recovery of imFASP method for trace amounts of complex proteome samples. All these results demonstrate that the imFASP method developed here is of high potential for high efficient and high throughput preparation of trace amounts of complex proteome samples.

  2. Porous texture of activated carbons prepared by phosphoric acid activation of woods

    NASA Astrophysics Data System (ADS)

    Díaz-Díez, M. A.; Gómez-Serrano, V.; Fernández González, C.; Cuerda-Correa, E. M.; Macías-García, A.

    2004-11-01

    Activated carbons (ACs) have been prepared using chestnut, cedar and walnut wood shavings from furniture industries located in the Comunidad Autónoma de Extremadura (SW Spain). Phosphoric acid (H3PO4) at different concentrations (i.e. 36 and 85 wt.%) has been used as activating agent. ACs have been characterized from the results obtained by N2 adsorption at 77 K. Moreover, the fractal dimension (D) has been calculated in order to determine the AC surface roughness degree. Optimal textural properties of ACs have been obtained by chemical activation with H3PO4 36 wt.%. This is corroborated by the slightly lower values of D for samples treated with H3PO4 85 wt.%.

  3. Preparation of layered thin film samples for angle-resolved photoemission spectroscopy

    SciTech Connect

    Harrison, S. E.; Zhou, B.; Huo, Y.; Harris, J. S.; Pushp, A.; Kellock, A. J.; Parkin, S. S. P.; Chen, Y.; Hesjedal, T.

    2014-09-22

    Materials with layered van der Waals crystal structures are exciting research topics in condensed matter physics and materials science due to outstanding physical properties associated with their strong two dimensional nature. Prominent examples include bismuth tritelluride and triselenide topological insulators (TIs), which are characterized by a bulk bandgap and pairwise counter-propagating spin-polarized electronic surface states. Angle-resolved photoemission spectroscopy (ARPES) of ex-situ grown thin film samples has been limited by the lack of suitable surface preparation techniques. We demonstrate the shortcomings of previously successful conventional surface preparation techniques when applied to ternary TI systems which are susceptible to severe oxidation. We show that in-situ cleaving is a simple and effective technique for preparation of clean surfaces on ex-situ grown thin films for high quality ARPES measurements. The method presented here is universally applicable to other layered van der Waals systems as well.

  4. Comparison of the effects of three different (-)-hydroxycitric acid preparations on food intake in rats: response

    PubMed Central

    Preuss, Harry G; Bagchi, Manashi; Bagchi, Debasis

    2006-01-01

    A response to Louter-van de Haar J, Wielinga PY, Scheurink AJ, Nieuwenhuizen AG: Comparison of the effects of three different (-)-hydroxycitric acid preparations on food intake in rats. Nutr Metabol 2005, 2:23 PMID:16846513

  5. Fuel properties of heptadecene isomers prepared via tandem isomerization-decarboxylation of oleic acid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Heptadecene isomers were prepared via tandem isomerization-decarboxylation of oleic acid using catalytic triruthenium dodecacarbonyl [Ru3(CO)12]. Chromatographic and spectroscopic characterization of the isolated heptadecene mixture indicated that it consisted of 96% internal isomers and 4% aromatic...

  6. The isomeric effect of mercaptobenzoic acids on the preparation and fluorescence properties of copper nanoclusters.

    PubMed

    Lin, Yi-Jyun; Chen, Po-Cheng; Yuan, Zhiqin; Ma, Jia-Ying; Chang, Huan-Tsung

    2015-08-04

    A one-pot approach has been developed to synthesize copper nanocluster (Cu NC) aggregates from copper nitrate and mercaptobenzoic acid (MBA). Cu NCs prepared separately in the three isomers of MBA exhibit different physical and optical properties.

  7. Capillary Electrophoresis Analysis of Organic Amines and Amino Acids in Saline and Acidic Samples Using the Mars Organic Analyzer

    NASA Astrophysics Data System (ADS)

    Stockton, Amanda M.; Chiesl, Thomas N.; Lowenstein, Tim K.; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A.

    2009-11-01

    The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pKa values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

  8. Capillary electrophoresis analysis of organic amines and amino acids in saline and acidic samples using the Mars organic analyzer.

    PubMed

    Stockton, Amanda M; Chiesl, Thomas N; Lowenstein, Tim K; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A

    2009-11-01

    The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pK(a) values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

  9. On the use of ultracentrifugal devices for routine sample preparation in biomolecular magic-angle-spinning NMR.

    PubMed

    Mandal, Abhishek; Boatz, Jennifer C; Wheeler, Travis B; van der Wel, Patrick C A

    2017-02-22

    A number of recent advances in the field of magic-angle-spinning (MAS) solid-state NMR have enabled its application to a range of biological systems of ever increasing complexity. To retain biological relevance, these samples are increasingly studied in a hydrated state. At the same time, experimental feasibility requires the sample preparation process to attain a high sample concentration within the final MAS rotor. We discuss these considerations, and how they have led to a number of different approaches to MAS NMR sample preparation. We describe our experience of how custom-made (or commercially available) ultracentrifugal devices can facilitate a simple, fast and reliable sample preparation process. A number of groups have since adopted such tools, in some cases to prepare samples for sedimentation-style MAS NMR experiments. Here we argue for a more widespread adoption of their use for routine MAS NMR sample preparation.

  10. Should the mass of a nanoferrite sample prepared by autocombustion method be considered as a realistic preparation parameter?

    NASA Astrophysics Data System (ADS)

    Wahba, Adel Maher; Mohamed, Mohamed Bakr

    2017-02-01

    Detectable variations in structural, elastic and magnetic properties have been reported depending on the mass of the cobalt nanoferrite sample prepared by citrate autocombustion method. Heat released during the autocombustion process and its duration are directly proportional to the mass to be prepared, and is thus expected to affect both the crystallite size and the cation distribution giving rise to the reported variations in microstrain, magnetization, and coercivity. Formation of a pure spinel phase has been validated using X-ray diffraction patterns (XRD) and Fourier-transform infrared (FTIR) spectra. Crystallite sizes obtained from Williamson-Hall (W-H) method range from 28-87 nm, being further supported by images of high-resolution transmission electron microscope (HRTEM). Saturation magnetization and coercivity deduced from M-H hysteresis loops show a clear correlation with the cation distribution, which was proposed on the basis of experimentally obtained data of XRD, VSM, and IR. Elastic parameters have been estimated using the cation distribution and FTIR data, with a resulting trend quite opposite to that of the lattice parameter.

  11. An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology

    PubMed Central

    Thompson, Rebecca F.; Walker, Matt; Siebert, C. Alistair; Muench, Stephen P.; Ranson, Neil A.

    2016-01-01

    Transmission electron microscopy (EM) is a versatile technique that can be used to image biological specimens ranging from intact eukaryotic cells to individual proteins >150 kDa. There are several strategies for preparing samples for imaging by EM, including negative staining and cryogenic freezing. In the last few years, cryo-EM has undergone a ‘resolution revolution’, owing to both advances in imaging hardware, image processing software, and improvements in sample preparation, leading to growing number of researchers using cryo-EM as a research tool. However, cryo-EM is still a rapidly growing field, with unique challenges. Here, we summarise considerations for imaging of a range of specimens from macromolecular complexes to cells using EM. PMID:26931652

  12. Optimization of proteomic sample preparation procedures for comprehensive protein characterization of pathogenic systems

    SciTech Connect

    Brewer, Heather M.; Norbeck, Angela D.; Adkins, Joshua N.; Manes, Nathan P.; Ansong, Charles; Shi, Liang; Rikihisa, Yasuko; Kikuchi, Takane; Wong, Scott; Estep, Ryan D.; Heffron, Fred; Pasa-Tolic, Ljiljana; Smith, Richard D.

    2008-12-19

    The elucidation of critical functional pathways employed by pathogens and hosts during an infectious cycle is both challenging and central to our understanding of infectious diseases. In recent years, mass spectrometry-based proteomics has been used as a powerful tool to identify key pathogenesis-related proteins and pathways. Despite the analytical power of mass spectrometry-based technologies, samples must be appropriately prepared to characterize the functions of interest (e.g. host-response to a pathogen or a pathogen-response to a host). The preparation of these protein samples requires multiple decisions about what aspect of infection is being studied, and it may require the isolation of either host and/or pathogen cellular material.

  13. Bio-sample preparation and gas chromatographic determination of benzodiazepines--a review.

    PubMed

    Uddin, Mohammad Nasir; Samanidou, Victoria F; Papadoyannis, Ioannis N

    2013-08-01

    Benzodiazepines have become commonly prescribed medicines worldwide in the therapy of anxiety, sleep disorders and convulsive attacks because they are relatively safe, with mild side effects. The availability of rapid, sensitive and selective analytical methods is essential for the determination of these drugs in clinical and forensic cases. Benzodiazepines are usually present at trace levels (μg/mL or ng/mL) in a complex biological matrix, and the potentially interfering compounds need to be removed before analysis. Therefore, a sample preparation technique is often mandatory, both to extract the drugs of interest from the matrices and to increase their concentration. An extended and comprehensive review is presented herein, focusing on bio-sample preparation (pretreatment, extraction and derivatization) and gas chromatographic methods applied for the quantification of 1,4-benzodiazepines.

  14. New sample preparation for quantitative laser desorption mass spectrometry and optical spectroscopy

    NASA Astrophysics Data System (ADS)

    Haefliger, Olivier P.; Zenobi, Renato

    1998-04-01

    Several analytical mass spectrometric and optical spectroscopic methods require a step during which a nonvolatile substance is desorbed by a laser pulse. It is, however, very difficult to use these methods for quantitative measurements because an accurate control over the amount desorbed by the laser pulse is generally not possible, especially when mixtures of several substances are used. We report a new fast and convenient sample preparation procedure that solves these problems. A solution of the analytes is mixed with a solution of poly(vinyl chloride) to obtain a homogeneous and vacuum-stable thin polymer membrane after the solvent has evaporated. Laser ablation is then performed directly from this membrane, allowing an accurate control of the amount of ablated analytes and excellent reproducibility. Quantitative laser desorption mass spectrometry over three orders of magnitude as well as optical spectroscopic measurements using this sample preparation method are demonstrated for polycyclic aromatic hydrocarbons.

  15. Bio-sample preparation and analytical methods for the determination of tricyclic antidepressants.

    PubMed

    Uddin, Mohammad N; Samanidou, Victoria F; Papadoyannis, Ioannis N

    2011-01-01

    An extended and comprehensive review is presented herein, focusing on sample preparation (pretreatment and extraction) and different analytical methods applied for the quantification of tricyclic antidepressants. These procedures are relevant tools in clinical and forensic toxicology. It is revealed that SPE, for sample preparation, and HPLC, using reversed-phase alkyl (C18) or cyanopropyl-bonded silica columns for the analytes separation, are effective and versatile methods for assay of tricyclic antidepressants. These methods enable achievable detection limits using UV/diode array detection, readily available in most laboratories, down to 1-8 ng ml(-1), and using electron capture detection better than 1 ng ml(-1), which is lower than that for nitrogen-phosphorus detector. MS interfaced with electrospray ionization offered similar sensitivity, whilst sonic spray ionization provided detection down to 0.03 ng ml(-1). A brief discussion on chemical structures, metabolism and mechanism of action of this group of drugs is also presented.

  16. SRAT CHEMISTRY AND ACID CONSUMPTION DURING SIMULATED DWPF MELTER FEED PREPARATION

    SciTech Connect

    Koopman, D; David Best, D; Bradley Pickenheim, B

    2008-12-03

    Due to higher than expected hydrogen generation during the Tank 51-Sludge Batch 4 (SB4) qualification run, DWPF engineering requested the Savannah River National Laboratory (SRNL) to expand the ongoing catalytic hydrogen generation program. The work presented in this Technical Report was identified as part of SRNL/Liquid Waste Organization (LWO) meetings to define potential causes of catalytic hydrogen generation as well as from an external technical review panel commissioned to evaluate SRNL hydrogen related data and programs. New scope included improving the understanding of SRAT/SME process chemistry, particularly as it related to acid consumption and hydrogen generation. The expanded hydrogen program scope was covered under the technical task request (TTR): HLW-DWPF-TTR-2007-0016. A task technical and quality assurance plan (TT&QAP) was issued to cover focus areas raised in meetings with LWO plus a portion of the recommendations made by the review panel. A supporting analytical study plan was issued. It was also noted in the review of catalytic hydrogen generation that control of the DWPF acid stoichiometry was an important element in controlling hydrogen generation. A separate TTR was issued to investigate ways of improving the determination of the acid requirement during processing: HLWDWPF-TTR-0015. A separate TT&QAP was prepared for this task request. This report discusses some progress on this task related to developing alternative acid equations and to performing experimental work to supplement the existing database. Simulant preparation and preliminary flowsheet studies were already documented. The prior work produced a sufficient quantity of simulant for the hydrogen program and melter feed rheology testing. It also defined a suitable acid addition stoichiometry. The results presented in this report come from samples and process data obtained during sixteen 22-L SRAT/SME simulations that were performed in the second half of 2007 to produce eight SME

  17. Influence of x-ray diffraction sample preparation on quantitative mineralogy: implications for chromate waste treatment.

    PubMed

    Dermatas, Dimitris; Chrysochoou, Maria; Pardali, Sarra; Grubb, Dennis G

    2007-01-01

    Powders of chromite ore processing residue (COPR) were mineralogically evaluated using quantitative X-ray powder diffraction (XRPD) to illustrate the impacts of sample preparation procedures. Chromite ore processing residue is strongly alkaline, reactive, contains minerals of varying hardness and absorption coefficients, and exhibits significant amorphicity. This poses a challenge to produce powders for XRPD analysis that are sufficiently fine and of uniform particle size while avoiding mineral reactions and overgrinding effects. Dry, hand pulverization to different grain sizes, and wet, mechanical pulverization (micromilling) using four milling liquids (cyclohexane, isopropanol, ethanol, and water), and variable milling durations (up to 15 min) were evaluated. Micromilling with a light, nonpolar, highly evaporative liquid such as cyclohexane with a milling time of 5 min mitigated systematic errors such as microabsorption and preferred orientation as it produced finer and more uniform particle size distributions than the hand-pulverized powders, while simultaneously affording the least time for sample preparation. Conversely, the use of water as milling liquid resulted in extensive hydration reactions during sample preparation, causing mischaracterization and significant underestimation of its reactive brownmillerite content, which can complicate the remediation design process for COPR. Hand pulverization emerged as a necessary complement to quantify Cr(VI)-containing, softer minerals destroyed during mechanical milling, the quantification of which has also important implications for COPR treatment design. The findings of this study may be applicable in a variety of geochemically complicated and reactive environmental media (metal-contaminated soils, stabilized/solidified media, inorganic waste), and points to the importance of the sample preparation method to obtain reliable quantitative XRPD results.

  18. Development of an automated sample preparation module for environmental monitoring of biowarfare agents.

    PubMed

    Hindson, Benjamin J; Brown, Steve B; Marshall, Graham D; McBride, Mary T; Makarewicz, Anthony J; Gutierrez, Dora M; Wolcott, Duane K; Metz, Thomas R; Madabhushi, Ramakrishna S; Dzenitis, John M; Colston, Billy W

    2004-07-01

    An automated sample preparation module, based upon sequential injection analysis (SIA), has been developed for use within an autonomous pathogen detection system. The SIA system interfaced aerosol sampling with multiplexed microsphere immunoassay-flow cytometric detection. Metering and sequestering of microspheres using SIA was found to be reproducible and reliable, over 24-h periods of autonomous operation. Four inbuilt immunoassay controls showed excellent immunoassay and system stability over five days of unattended continuous operation. Titration curves for two biological warfare agents, Bacillus anthracis and Yersinia pestis, obtained using the automated SIA procedure were shown to be similar to those generated using a manual microtiter plate procedure.

  19. TMT One-Stop Shop: From Reliable Sample Preparation to Computational Analysis Platform.

    PubMed

    Mirzaei, Mehdi; Pascovici, Dana; Wu, Jemma X; Chick, Joel; Wu, Yunqi; Cooke, Brett; Haynes, Paul; Molloy, Mark P

    2017-01-01

    In this chapter we describe the workflow we use for labeled quantitative proteomics analysis using tandem mass tags (TMT) starting with the sample preparation and ending with the multivariate analysis of the resulting data. We detail the step-by-step process from sample processing, labeling, fractionation, and data processing using Proteome Discoverer through to data analysis and interpretation in the context of a multi-run experiment. The final analysis and data interpretation rely on an R package we call TMTPrepPro, which are deployed on a local GenePattern server, and used for generating various outputs which are also outlined herein.

  20. Optimization of Proteomic Sample Preparation Procedures for Comprehensive Protein Characterization of Pathogenic Systems

    PubMed Central

    Mottaz-Brewer, Heather M.; Norbeck, Angela D.; Adkins, Joshua N.; Manes, Nathan P.; Ansong, Charles; Shi, Liang; Rikihisa, Yasuko; Kikuchi, Takane; Wong, Scott W.; Estep, Ryan D.; Heffron, Fred; Pasa-Tolic, Ljiljana; Smith, Richard D.

    2008-01-01

    Mass spectrometry-based proteomics is a powerful analytical tool for investigating pathogens and their interactions within a host. The sensitivity of such analyses provides broad proteome characterization, but the sample-handling procedures must first be optimized to ensure compatibility with the technique and to maximize the dynamic range of detection. The decision-making process for determining optimal growth conditions, preparation methods, sample analysis methods, and data analysis techniques in our laboratory is discussed herein with consideration of the balance in sensitivity, specificity, and biomass losses during analysis of host-pathogen systems. PMID:19183792

  1. Preparation of polyethylene sacks for collection of precipitation samples for chemical analysis

    USGS Publications Warehouse

    Schroder, L.J.; Bricker, A.W.

    1985-01-01

    Polyethylene sacks are used to collect precipitation samples. Washing polyethylene with acetone, hexane, methanol, or nitric acid can change the adsorptive characteristics of the polyethylene. In this study, simulated precipitation at pH 4.5 was in contact with the polyethylene sacks for 21 days; subsamples were removed for chemical analysis at 7, 14, and 21 days after intitial contact. Sacks washed with acetone adsorbed iron and lithium; sacks washed with hexane adsorbed barium, iron , and lithium; sacks washed with methanol adsorbed calcium and iron; and sacks washed with 0.30 N nitric acid adsorbed iron. Leaching the plastic sacks with 0.15 N nitric acid did not result in 100-percent recovery of any of the adsorbed metals. Washing polyethylene sacks with dilute nitric acid caused the pH of the simulated precipitation to be decreased by 0.2 pH unit after 1 week of contact with the polyethylene. The specific conductance increased by 10 microsiemens per centimeter. Contamination of precipitation samples by lead was determined to be about 0.1 microgram per liter from contact with precleaned polyethylene sacks. No measurable contamination of precipitation samples by zinc occurred. (USGS)

  2. Simple Sodium Dodecyl Sulfate-Assisted Sample Preparation Method for LC-MS-based Proteomic Applications

    SciTech Connect

    Zhou, Jianying; Dann, Geoffrey P.; Shi, Tujin; Wang, Lu; Gao, Xiaoli; Su, Dian; Nicora, Carrie D.; Shukla, Anil K.; Moore, Ronald J.; Liu, Tao; Camp, David G.; Smith, Richard D.; Qian, Weijun

    2012-03-10

    Sodium dodecyl sulfate (SDS) is one of the most popular laboratory reagents used for highly efficient biological sample extraction; however, SDS presents a significant challenge to LC-MS-based proteomic analyses due to its severe interference with reversed-phase LC separations and electrospray ionization interfaces. This study reports a simple SDS-assisted proteomic sample preparation method facilitated by a novel peptide-level SDS removal protocol. After SDS-assisted protein extraction and digestion, SDS was effectively (>99.9%) removed from peptides through ion substitution-mediated DS- precipitation with potassium chloride (KCl) followed by {approx}10 min centrifugation. Excellent peptide recovery (>95%) was observed for less than 20 {mu}g of peptides. Further experiments demonstrated the compatibility of this protocol with LC-MS/MS analyses. The resulting proteome coverage from this SDS-assisted protocol was comparable to or better than those obtained from other standard proteomic preparation methods in both mammalian tissues and bacterial samples. These results suggest that this SDS-assisted protocol is a practical, simple, and broadly applicable proteomic sample processing method, which can be particularly useful when dealing with samples difficult to solubilize by other methods.

  3. DNA isolation and sample preparation for quantification of adduct levels by accelerator mass spectrometry.

    PubMed

    Dingley, Karen H; Ubick, Esther A; Vogel, John S; Ognibene, Ted J; Malfatti, Michael A; Kulp, Kristen; Haack, Kurt W

    2014-01-01

    Accelerator mass spectrometry (AMS) is a highly sensitive technique used for the quantification of adducts following exposure to carbon-14- or tritium-labeled chemicals, with detection limits in the range of one adduct per 10(11)-10(12) nucleotides. The protocol described in this chapter provides an optimal method for isolating and preparing DNA samples to measure isotope-labeled DNA adducts by AMS. When preparing samples, special precautions must be taken to avoid cross-contamination of isotope among samples and produce a sample that is compatible with AMS. The DNA isolation method described is based upon digestion of tissue with proteinase K, followed by extraction of DNA using Qiagen isolation columns. The extracted DNA is precipitated with isopropanol, washed repeatedly with 70 % ethanol to remove salt, and then dissolved in water. DNA samples are then converted to graphite or titanium hydride and the isotope content measured by AMS to quantify adduct levels. This method has been used to reliably generate good yields of uncontaminated, pure DNA from animal and human tissues for analysis of adduct levels.

  4. Sample preparation for combined chemical analysis and in vitro bioassay application in water quality assessment.

    PubMed

    Kolkman, Annemieke; Schriks, Merijn; Brand, Walter; Bäuerlein, Patrick S; van der Kooi, Margaretha M E; van Doorn, René H; Emke, Erik; Reus, Astrid A; van der Linden, Sander C; de Voogt, Pim; Heringa, Minne B

    2013-11-01

    The combination of in vitro bioassays and chemical screening can provide a powerful toolbox to determine biologically relevant compounds in water extracts. In this study, a sample preparation method is evaluated for the suitability for both chemical analysis and in vitro bioassays. A set of 39 chemicals were spiked to surface water, which were extracted using Oasis MCX cartridges. The extracts were chemically analyzed by liquid chromatography linear ion trap Orbitrap analysis and recoveries appeared to be on average 61% Compounds with logK(ow) values in the range between 0 and 4 are recovered well using this method. In a next step, the same extracts were tested for genotoxic activity using the Comet assay and Ames fluctuation test and for specific endocrine receptor activation using a panel of CALUX assays, for estrogenic (ER), androgenic (AR), glucocorticoid (GR), progestagenic (PR), and thyroidogenic (TR) agonistic activities. The results of the genotoxicity assays indicated that spiked genotoxic compounds were preserved during sample preparation. The measured responses of the GR CALUX and ER CALUX assays were similar to the predicted responses. The measured responses in the AR CALUX and PR CALUX assays were much lower than expected from the analytical concentration, probably due to antagonistic effects of some spiked compounds. Overall, the presented sample preparation method seems to be suitable for both chemical analysis and specific in vitro bioassay applications.

  5. Error baseline rates of five sample preparation methods used to characterize RNA virus populations.

    PubMed

    Kugelman, Jeffrey R; Wiley, Michael R; Nagle, Elyse R; Reyes, Daniel; Pfeffer, Brad P; Kuhn, Jens H; Sanchez-Lockhart, Mariano; Palacios, Gustavo F

    2017-01-01

    Individual RNA viruses typically occur as populations of genomes that differ slightly from each other due to mutations introduced by the error-prone viral polymerase. Understanding the variability of RNA virus genome populations is critical for understanding virus evolution because individual mutant genomes may gain evolutionary selective advantages and give rise to dominant subpopulations, possibly even leading to the emergence of viruses resistant to medical countermeasures. Reverse transcription of virus genome populations followed by next-generation sequencing is the only available method to characterize variation for RNA viruses. However, both steps may lead to the introduction of artificial mutations, thereby skewing the data. To better understand how such errors are introduced during sample preparation, we determined and compared error baseline rates of five different sample preparation methods by analyzing in vitro transcribed Ebola virus RNA from an artificial plasmid-based system. These methods included: shotgun sequencing from plasmid DNA or in vitro transcribed RNA as a basic "no amplification" method, amplicon sequencing from the plasmid DNA or in vitro transcribed RNA as a "targeted" amplification method, sequence-independent single-primer amplification (SISPA) as a "random" amplification method, rolling circle reverse transcription sequencing (CirSeq) as an advanced "no amplification" method, and Illumina TruSeq RNA Access as a "targeted" enrichment method. The measured error frequencies indicate that RNA Access offers the best tradeoff between sensitivity and sample preparation error (1.4-5) of all compared methods.

  6. Sample preparation: a critical step in the analysis of cholesterol oxidation products.

    PubMed

    Georgiou, Christiana A; Constantinou, Michalis S; Kapnissi-Christodoulou, Constantina P

    2014-02-15

    In recent years, cholesterol oxidation products (COPs) have drawn scientific interest, particularly due to their implications on human health. A big number of these compounds have been demonstrated to be cytotoxic, mutagenic, and carcinogenic. The main source of COPs is through diet, and particularly from the consumption of cholesterol-rich foods. This raises questions about the safety of consumers, and it suggests the necessity for the development of a sensitive and a reliable analytical method in order to identify and quantify these components in food samples. Sample preparation is a necessary step in the analysis of COPs in order to eliminate interferences and increase sensitivity. Numerous publications have, over the years, reported the use of different methods for the extraction and purification of COPs. However, no method has, so far, been established as a routine method for the analysis of COPs in foods. Therefore, it was considered important to overview different sample preparation procedures and evaluate the different preparative parameters, such as time of saponification, the type of organic solvents for fat extraction, the stationary phase in solid phase extraction, etc., according to recovery, precision and simplicity.

  7. Preparation of novel polydimethylsiloxane solid-phase microextraction film and its application in liquid sample pretreatment.

    PubMed

    Wei, Fang; Zhang, Fang-Fang; Liao, Hui; Dong, Xu-Yan; Li, Yan-Hua; Chen, Hong

    2011-02-01

    In this paper, an extraction approach based on the use of a novel polydimethylsiloxane (PDMS) film as the extraction medium was described. Two kinds of PDMS films with smooth surface and frosted surface were prepared and were practically evaluated for extraction. A model analytical problem, the determination of organochlorine pesticides in water samples, was selected for practical evaluation of the film extraction procedure by direct extraction and solvent desorption, followed by gas chromatography (GC) analysis with electron capture detection (ECD). The main variables affecting the extraction process such as the extraction time, the extraction temperature, the elution conditions, etc. were studied. The method was characterized on the basis of its linearity, precision, and limits of detection. The novel approach was sensitive and precise enough for the detection of the target analytes in the low nanogram per liter range using 5 mL of sample. In fact, limits of detection ranging from 0.77 to 10.25 ng/L were obtained. Compared with the solid-phase microextraction (SPME) fiber, the robust extraction film has a large extraction capacity, low cost of preparation. Besides, owing to the simplicity of the extraction procedures, in-site sample preparation for environmental monitor may be realized.

  8. A fully automated plasma protein precipitation sample preparation method for LC-MS/MS bioanalysis.

    PubMed

    Ma, Ji; Shi, Jianxia; Le, Hoa; Cho, Robert; Huang, Judy Chi-jou; Miao, Shichang; Wong, Bradley K

    2008-02-01

    This report describes the development and validation of a robust robotic system that fully integrates all peripheral devices needed for the automated preparation of plasma samples by protein precipitation. The liquid handling system consisted of a Tecan Freedom EVO 200 liquid handling platform equipped with an 8-channel liquid handling arm, two robotic plate-handling arms, and two plate shakers. Important additional components integrated into the platform were a robotic temperature-controlled centrifuge, a plate sealer, and a plate seal piercing station. These enabled unattended operation starting from a stock solution of the test compound, a set of test plasma samples and associated reagents. The stock solution of the test compound was used to prepare plasma calibration and quality control samples. Once calibration and quality control samples were prepared, precipitation of plasma proteins was achieved by addition of three volumes of acetonitrile. Integration of the peripheral devices allowed automated sequential completion of the centrifugation, plate sealing, piercing and supernatant transferral steps. The method produced a sealed, injection-ready 96-well plate of plasma extracts. Accuracy and precision of the automated system were satisfactory for the intended use: intra-day and the inter-day precision were excellent (C.V.<5%), while the intra-day and inter-day accuracies were acceptable (relative error<8%). The flexibility of the platform was sufficient to accommodate pharmacokinetic studies of different numbers of animals and time points. To the best of our knowledge, this represents the first complete automation of the protein precipitation method for plasma sample analysis.

  9. [New synthesis of the anticoagulant pentasaccharide idraparinux and preparation of its analogues containing sulfonic acid moieties].

    PubMed

    Herczeg, Mihály

    2012-01-01

    Two novel synthetic pathways were elaborated for the preparation of idraparinux, a heparin-related fully O-sulfated, O-methylated anticoagulant pentasaccharide. Both methods based upon a [2+3] block synthesis utilizing the same trisaccharide acceptor which was coupled to either a uronic acid disaccharide donor or its nonoxidized precursor. Two bioisosteric sulfonic acid analogues of idraparinux were also prepared, in which two or three primary sulfate esters were replaced by sodium-sulfonatomethyl moieties. The sulfonic acid groups were formed on a monosaccharide level and the obtained carbohydrate sulfonic acid esters were found to be excellent donors and acceptors in the glycosylation reactions. The disulfonic-acid analogue was prepared in a [2+3] block synthesis by using a trisaccharide disulfonic acid as an acceptor and a glucuronide disaccharide as a donor. For the synthesis of the pentasaccharide trisulfonic acid, a more-efficient approach, which involved elongation of the trisaccharide acceptor with a non-oxidized precursor of the glucuronic acid followed by post-glycosidation oxidation at the tetrasaccharide level and a subsequent [1+4] coupling reaction, was elaborated. In vitro evaluation of the anticoagulant activity of the reference compound idraparinux and the new sulfonic acid derivatives revealed that the disulfonate analogue inhibited the blood-coagulation-proteinase factor Xa with outstanding efficacy; however, the introduction of the third sulfonic acid moiety resulted in a notable decrease in the anti-Xa activity.

  10. Lab-on-a-chip modules for detection of highly pathogenic bacteria: from sample preparation to detection

    NASA Astrophysics Data System (ADS)

    Julich, S.; Kopinč, R.; Hlawatsch, N.; Moche, C.; Lapanje, A.; Gärtner, C.; Tomaso, H.

    2014-05-01

    Lab-on-a-chip systems are innovative tools for the detection and identification of microbial pathogens in human and veterinary medicine. The major advantages are small sample volume and a compact design. Several fluidic modules have been developed to transform analytical procedures into miniaturized scale including sampling, sample preparation, target enrichment, and detection procedures. We present evaluation data for single modules that will be integrated in a chip system for the detection of pathogens. A microfluidic chip for purification of nucleic acids was established for cell lysis using magnetic beads. This assay was evaluated with spiked environmental aerosol and swab samples. Bacillus thuringiensis was used as simulant for Bacillus anthracis, which is closely related but non-pathogenic for humans. Stationary PCR and a flow-through PCR chip module were investigated for specific detection of six highly pathogenic bacteria. The conventional PCR assays could be transferred into miniaturized scale using the same temperature/time profile. We could demonstrate that the microfluidic chip modules are suitable for the respective purposes and are promising tools for the detection of bacterial pathogens. Future developments will focus on the integration of these separate modules to an entire lab-on-a-chip system.

  11. Improved sample preparation and counting techniques for enhanced tritium measurement sensitivity

    NASA Astrophysics Data System (ADS)

    Moran, J.; Aalseth, C.; Bailey, V. L.; Mace, E. K.; Overman, C.; Seifert, A.; Wilcox Freeburg, E. D.

    2015-12-01

    Tritium (T) measurements offer insight to a wealth of environmental applications including hydrologic tracking, discerning ocean circulation patterns, and aging ice formations. However, the relatively short half-life of T (12.3 years) limits its effective age dating range. Compounding this limitation is the decrease in atmospheric T content by over two orders of magnitude (from 1000-2000 TU in 1962 to < 10 TU currently) since the cessation of above ground nuclear testing in the 1960's. We are developing sample preparation methods coupled to direct counting of T via ultra-low background proportional counters which, when combined, offer improved T measurement sensitivity (~4.5 mmoles of H2 equivalent) and will help expand the application of T age dating to smaller sample sizes linked to persistent environmental questions despite the limitations above. For instance, this approach can be used to T date ~ 2.2 mmoles of CH4 collected from sample-limited systems including microbial communities, soils, or subsurface aquifers and can be combined with radiocarbon dating to distinguish the methane's formation age from C age in a system. This approach can also expand investigations into soil organic C where the improved sensitivity will permit resolution of soil C into more descriptive fractions and provide direct assessments of the stability of specific classes of organic matter in soils environments. We are employing a multiple step sample preparation system whereby organic samples are first combusted with resulting CO2 and H2O being used as a feedstock to synthesize CH4. This CH4 is mixed with Ar and loaded directly into an ultra-low background proportional counter for measurement of T β decay in a shallow underground laboratory. Analysis of water samples requires only the addition of geologic CO2 feedstock with the sample for methane synthesis. The chemical nature of the preparation techniques enable high sample throughput with only the final measurement requiring T decay

  12. Rapid Measurement of Food Adulteration with Minimal Sample Preparation and No Chromatography Using Ambient Ionization Mass Spectrometry.

    PubMed

    Dalmia, Avinash

    2017-03-01

    A rapid method, with minimal sample preparation and no chromatography, was developed for analyzing food samples such as olive oil and pomegranate juice to measure adulteration with cheaper ingredients using the novel Direct Sample Analysis™ (DSA) ion source in conjunction with a time-of-flight (TOF)-MS. In less than 30 s, with minimal sample preparation and method development, adulteration of olive oil and pomegranate juice with cheaper seed oils and fruit juices, respectively, was measured with DSA/TOF-MS.

  13. Preparation of Saturated and Unsaturated Fatty Acid Hydrazides and Long Chain C-glycoside Ketohydrazones

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A method is described to prepare both saturated and unsaturated fatty acid acyl hydrazides using lipase as a catalyst. Hydrazides were generated from fatty acid methyl esters as well as directly from vegetable oils, and an organic co-solvent was not needed to maintain the integrity of the unsaturat...

  14. Preparation of 4-amino-2,4-dioxobutanoic acid

    DOEpatents

    Unkefer, Pat J.; Martinez, Rodolfo A.; Glass, David R.

    2016-03-22

    A process for synthesizing 4-amino-2,4-dioxobutanoic acid involves reacting diethyl oxalate with an alkoxide in ethanol to form a reaction mixture, and afterward adding ethyl cyanoacetate to the reaction mixture and allowing a reaction to proceed under conditions suitable to form a first reaction product of the formula diethyl 2-cyano-3-hydroxy-butenedioate, and then isolating the diethyl 2-cyano-3-hydroxy-butenedioate, and afterward reacting the diethyl-2-cyano-3-hydroxy-butenedioate with an aqueous hydroxide under conditions suitable to form 4-amino-2,4-dioxobutanoic acid.

  15. Preparation of 4-amino-2,4-dioxobutanoic acid

    DOEpatents

    Unkefer, Pat J.; Martinez, Rodolfo A.; Glass, David R.

    2015-06-02

    A process for synthesizing 4-amino-2,4-dioxobutanoic acid involves reacting diethyl oxalate with sodium ethoxide in ethanol to form a reaction mixture, and afterward adding ethyl cyanoacetate to the reaction mixture and allowing a reaction to proceed under conditions suitable to form a first reaction product of the formula diethyl-2-cyano-3-hydroxy-butenedioate, and then isolating the diethyl-2-cyano-3-hydroxybutenedioate, and afterward reacting the diethyl-2-cyano-3-hydroxy-butenedioate with aqueous sodium hydroxide under conditions suitable to form 4-amino-2,4-dioxobutanoic acid.

  16. Controlled antibody release from gelatin for on-chip sample preparation.

    PubMed

    Zhang, Xichen; Wasserberg, Dorothee; Breukers, Christian; Terstappen, Leon W M M; Beck, Markus

    2016-05-10

    A practical way to realize on-chip sample preparation for point-of-care diagnostics is to store the required reagents on a microfluidic device and release them in a controlled manner upon contact with the sample. For the development of such diagnostic devices, a fundamental understanding of the release kinetics of reagents from suitable materials in microfluidic chips is therefore essential. Here, we study the release kinetics of fluorophore-conjugated antibodies from (sub-) μm thick gelatin layers and several ways to control the release time. The observed antibody release is well-described by a diffusion model. Release times ranging from ∼20 s to ∼650 s were determined for layers with thicknesses (in the dry state) between 0.25 μm and 1.5 μm, corresponding to a diffusivity of 0.65 μm(2) s(-1) (in the swollen state) for our standard layer preparation conditions. By modifying the preparation conditions, we can influence the properties of gelatin to realize faster or slower release. Faster drying at increased temperatures leads to shorter release times, whereas slower drying at increased humidity yields slower release. As expected in a diffusive process, the release time increases with the size of the antibody. Moreover, the ionic strength of the release medium has a significant impact on the release kinetics. Applying these findings to cell counting chambers with on-chip sample preparation, we can tune the release to control the antibody distribution after inflow of blood in order to achieve homogeneous cell staining.

  17. Soybean and Lactose in Meat Products and Preparations Sampled at Retail

    PubMed Central

    Piccolo, Filomena; Vollano, Lucia; Base, Giuseppe; Girasole, Mariagrazia; Smaldone, Giorgio; Cortesi, Maria Luisa

    2016-01-01

    Food allergies and intolerances have increased during the last decades and regulatory authorities have taken different measures to prevent and manage consumers’ adverse reactions, including correct labelling of foods. Aim of this work was to search for soybean and lactose in meat products and meat preparations taken from retail in some provinces of Campania Region (Southern Italy) and to evaluate the food labels compliance with Regulation (EU) n.1169/2011. Soybean and lactose were searched using commercial kits in n. 58 samples of meat products produced in or distributed by 19 establishments, and in n. 55 samples of meat products and n. 8 of meat preparations produced in 21 plants. All samples were selected on the basis of the absence of any information on the labels about the presence of the two searched allergens, with the exception of n. 5 samples tested for lactose. Traces of soybean were detected in 50 out of the 58 examined samples, at concentrations up to 0.93 mg kg–1. Only two samples contained levels above the detection limit of 0.31 mg kg–1. Lactose levels ranging from 0.11 to 2.95 g/100 g, i.e. above the detection limit, were found in all the tested samples (n. 63). The results of the present research underline the need for careful controls and planning by operators as part of the self-control plans, and deserve attention from the competent authorities considering not only the consumers’ health but also the great attention media pay to regulations providing consumers with information on food. PMID:27853709

  18. Soybean and Lactose in Meat Products and Preparations Sampled at Retail.

    PubMed

    Piccolo, Filomena; Vollano, Lucia; Base, Giuseppe; Girasole, Mariagrazia; Smaldone, Giorgio; Cortesi, Maria Luisa

    2016-06-03

    Food allergies and intolerances have increased during the last decades and regulatory authorities have taken different measures to prevent and manage consumers' adverse reactions, including correct labelling of foods. Aim of this work was to search for soybean and lactose in meat products and meat preparations taken from retail in some provinces of Campania Region (Southern Italy) and to evaluate the food labels compliance with Regulation (EU) n.1169/2011. Soybean and lactose were searched using commercial kits in n. 58 samples of meat products produced in or distributed by 19 establishments, and in n. 55 samples of meat products and n. 8 of meat preparations produced in 21 plants. All samples were selected on the basis of the absence of any information on the labels about the presence of the two searched allergens, with the exception of n. 5 samples tested for lactose. Traces of soybean were detected in 50 out of the 58 examined samples, at concentrations up to 0.93 mg kg(-1). Only two samples contained levels above the detection limit of 0.31 mg kg(-1). Lactose levels ranging from 0.11 to 2.95 g/100 g, i.e. above the detection limit, were found in all the tested samples (n. 63). The results of the present research underline the need for careful controls and planning by operators as part of the self-control plans, and deserve attention from the competent authorities considering not only the consumers' health but also the great attention media pay to regulations providing consumers with information on food.

  19. Membrane-based sample preparation for ion chromatography-Techniques, instrumental configurations and applications.

    PubMed

    Frenzel, Wolfgang; Markeviciute, Inga

    2017-01-06

    Sample preparation is the bottleneck of many analytical methods, including ion chromatography (IC). Procedures based on the application of membranes are important, yet not well appreciated means for clean-up and analyte preconcentration of liquid samples. Filtration, ultrafiltration, the variety of dialysis techniques, i.e. passive dialysis, Donnan dialysis and electrodialysis, as well as gas-diffusion are being reviewed here with respect to their application in combination with IC. Instrumental aspects including hardware requirements, configuration of membrane separation units and membrane characteristics are presented. Operation in batch and flow-through mode is described with emphasis on the latter to in-line coupling with IC, permitting fully automated operation. Attention is also drawn to dialysis probes and microdialysis both providing options for in-situ measurements with inherent selective sampling of analytes and sample preparation. The respective features of the various techniques are outlined with respect to the possibilities of matrix removal and selectivity enhancement. In this article, we provide examples of application of the diverse membrane separation techniques and discuss the benefits and limitations thereof.

  20. Sample preparation for precise and quantitative electron holographic analysis of semiconductor devices.

    PubMed

    Han, Myung-Geun; Li, Jing; Xie, Qianghua; Fejes, Peter; Conner, James; Taylor, Bill; McCartney, Martha R

    2006-08-01

    Wedge polishing was used to prepare one-dimensional Si n-p junction and Si p-channel metal-oxide-silicon field effect transistor (pMOSFET) samples for precise and quantitative electrostatic potential analysis using off-axis electron holography. To avoid artifacts associated with ion milling, cloth polishing with 0.02-microm colloidal silica suspension was used for final thinning. Uniform thickness and no significant charging were observed by electron holography analysis for samples prepared entirely by this method. The effect of sample thickness was investigated and the minimum thickness for reliable results was found to be approximately 160 nm. Below this thickness, measured phase changes were smaller than expected. For the pMOSFET sample, quantitative analysis of two-dimensional electrostatic potential distribution showed that the metallurgical gate length (separation between two extension junctions) was approximately 54 nm, whereas the actual gate length was measured to be approximately 70 nm by conventional transmission electron microscopy. Thus, source and drain junction encroachment under the gate was 16 nm.

  1. Sample preparation and direct electrospray ionization on a tip column for rapid mass spectrometry analysis of complex samples.

    PubMed

    Huang, Yun-Qing; You, Jin-Qing; Yuan, Bi-Feng; Feng, Yu-Qi

    2012-10-07

    A handheld pipette tip column electrospray ionization source (PTC-ESI source) was developed for rapid mass spectrometry analysis at ambient pressure. The PTC-ESI source was made up of three main component parts including a micro DC high voltage (HV) power supply, a micropipette and a disposable micropipette tip filled with a plug of adsorbent. A DC high voltage was applied to the sharp point of the micropipette tip column to induce electrospray ionization. The PTC-ESI source was successfully used for direct analysis of basic organic compounds, organic acids and peptides in a simple matrix. In the case of complex samples, micro-extraction based on the adsorbent phase filled in the pipette tip was used to remove impurities and concentrate target analytes prior to ionization. The eluting solution was not pipetted out, but directly dispersed in the form of electrospray from the pipette tip for ionization. The effectiveness of the PTC-ESI source has been further demonstrated by fast analysis of therapeutic compounds and endogenous bioactive chemicals in complex biological samples.

  2. Lead biomonitoring in different organs of lead intoxicated rats employing GF AAS and different sample preparations.

    PubMed

    de Sousa, Rafael Arromba; Sabarense, Céphora Maria; Prado, Gustavo L P; Metze, Konradin; Cadore, Solange

    2013-01-30

    An analytical procedure was developed for the determination of lead in different tissues from Wistar Hanover rats, previously intoxicated with lead acetate during a toxicological study. About 25 mg of dried sample (bone, liver, kidney, heart, lung and spleen) were mixed with 8.0 mL of 7.00 mol L(-1) nitric acid and digested using microwave radiation in closed vessel. Except for the bone samples, the other tissues could also be analyzed after alkaline solubilization with TMAH. All the digested or solubilized samples were analyzed by graphite furnace atomic absorption spectrometry. Good accuracy and precision were attained when analyzing reference standard materials (for bone, liver and kidney) and also from addition to recovery experiments (for heart, lung and spleen tissues). The method was applied to samples from nine animals and the results suggested that there is a profile for lead bioaccumulation in these animals, which seemed to adapt themselves to continuous lead exposure.

  3. Comparison of Different Sample Preparation Protocols Reveals Lysis Buffer-Specific Extraction Biases in Gram-Negative Bacteria and Human Cells.

    PubMed

    Glatter, Timo; Ahrné, Erik; Schmidt, Alexander

    2015-11-06

    We evaluated different in-solution and FASP-based sample preparation strategies for absolute protein quantification. Label-free quantification (LFQ) was employed to compare different sample preparation strategies in the bacterium Pseudomonas aeruginosa and human embryonic kidney cells (HEK), and organismal-specific differences in general performance and enrichment of specific protein classes were noted. The original FASP protocol globally enriched for most proteins in the bacterial sample, whereas the sodium deoxycholate in-solution strategy was more efficient with HEK cells. Although detergents were found to be highly suited for global proteome analysis, higher intensities were obtained for high-abundant nucleic acid-associated protein complexes, like the ribosome and histone proteins, using guanidine hydrochloride. Importantly, we show for the first time that the observable total proteome mass of a sample strongly depends on the sample preparation protocol, with some protocols resulting in a significant underestimation of protein mass due to incomplete protein extraction of biased protein groups. Furthermore, we demonstrate that some of the observed abundance biases can be overcome by incorporating a nuclease treatment step or, alternatively, a correction factor for complementary sample preparation approaches.

  4. Automated sample preparation facilitated by PhyNexus MEA purification system for oligosaccharide mapping of glycoproteins.

    PubMed

    Prater, Bradley D; Anumula, Kalyan R; Hutchins, Jeff T

    2007-10-15

    A reproducible high-throughput sample cleanup method for fluorescent oligosaccharide mapping of glycoproteins is described. Oligosaccharides are released from glycoproteins using PNGase F and labeled with 2-aminobenzoic acid (anthranilic acid, AA). A PhyNexus MEA system was adapted for automated isolation of the fluorescently labeled oligosaccharides from the reaction mixture prior to mapping by HPLC. The oligosaccharide purification uses a normal-phase polyamide resin (DPA-6S) in custom-made pipette tips. The resin volume, wash, and elution steps involved were optimized to obtain high recovery of oligosaccharides with the least amount of contaminating free fluorescent dye in the shortest amount of time. The automated protocol for sample cleanup eliminated all manual manipulations with a recycle time of 23 min. We have reduced the amount of excess AA by 150-fold, allowing quantitative oligosaccharide mapping from as little as 500 ng digested recombinant immunoglobulin G (rIgG). This low sample requirement allows early selection of a cell line with desired characteristics (e.g., oligosaccharide profile and high specific productivity) for the production of glycoprotein drugs. In addition, the use of Tecan or another robotic platform in conjunction with this method should allow the cleanup of 96 samples in 23 min, a significant decrease in the amount of time currently required to process such a large number of samples.

  5. Centrifugal concentrator for the substitution of nitrogen blow-down micro-concentration in dioxin/polychlorinated biphenyl sample preparation.

    PubMed

    Kemmochi, Yukio; Tsutsumi, Kaori; Arikawa, Akihiro; Nakazawa, Hiroyuki

    2002-01-18

    A centrifugal concentrator was applied for the substitution of nitrogen blow-down micro-concentration in dioxin/PCB sample preparation. Automation was achieved through this apparatus and resulted in significant improvements in sample preparation productivity, along with excellent reproducibility and recovery rate of dioxin/PCB. Mean recoveries of dioxin congeners were 90-106%, respectively.

  6. Preparation and evaluation of lignosulfonates as a dispersant for gypsum paste from acid hydrolysis lignin.

    PubMed

    Matsushita, Yasuyuki; Yasuda, Seiichi

    2005-03-01

    In order to effectively utilize a by-product of the acid saccharification process of woody materials, the chemical conversion of guaiacyl sulfuric acid lignin (SAL), one of the acid hydrolysis lignins, into water-soluble sulfonated products with high dispersibitity was investigated. At first, SAL was phenolated (P-SAL) to enhance the solubility and reactivity. Lignosulfonates were prepared from P-SAL by three methods of hydroxymethylation followed by neutral sulfonation (two-step method), sulfomethylation (one-step method) and arylsulfonation. Surprisingly, all prepared lignosulfonates possessed 30 to 70% higher dispersibility for gypsum paste than the commercial lignosulfonate. Evaluation of the preparations for gypsum paste suggested that the higher molecular weights and sulfur contents of the preparations increased their dispersibility.

  7. Process for the preparation of 3,4-dihydroxybutanoic acid and salts thereof

    DOEpatents

    Hollingsworth, Rawle I.

    1994-01-01

    A process for the preparation of 3,4-dihydroxybutanoic acid (1) and salts thereof from a glucose source containing 1,4-1inked glucose as a substituent is described. The process uses an alkali metal hdyroxide and hydrogen peroxide to convert the glucose source to (1). The compound (1) is useful as a chemical intermediate to naturally occurring fatty acids and is used to prepare 3,4-dihydroxybutanoic acid-gamma-lactone (2) and furanone (3), particularly stereoisomers of these compounds.

  8. Sources of variability in collection and preparation of paint and lead-coating samples.

    PubMed

    Harper, S L; Gutknecht, W F

    2001-06-01

    Chronic exposure of children to lead (Pb) can result in permanent physiological impairment. Since surfaces coated with lead-containing paints and varnishes are potential sources of exposure, it is extremely important that reliable methods for sampling and analysis be available. The sources of variability in the collection and preparation of samples were investigated to improve the performance and comparability of methods and to ensure that data generated will be adequate for its intended use. Paint samples of varying sizes (areas and masses) were collected at different locations across a variety of surfaces including metal, plaster, concrete, and wood. A variety of grinding techniques were compared. Manual mortar and pestle grinding for at least 1.5 min and mechanized grinding techniques were found to generate similar homogenous particle size distributions required for aliquots as small as 0.10 g. When 342 samples were evaluated for sample weight loss during mortar and pestle grinding, 4% had 20% or greater loss with a high of 41%. Homogenization and sub-sampling steps were found to be the principal sources of variability related to the size of the sample collected. Analysis of samples from different locations on apparently identical surfaces were found to vary by more than a factor of two both in Pb concentration (mg cm-2 or %) and areal coating density (g cm-2). Analyses of substrates were performed to determine the Pb remaining after coating removal. Levels as high as 1% Pb were found in some substrate samples, corresponding to more than 35 mg cm-2 Pb. In conclusion, these sources of variability must be considered in development and/or application of any sampling and analysis methodologies.

  9. A user-friendly robotic sample preparation program for fully automated biological sample pipetting and dilution to benefit the regulated bioanalysis.

    PubMed

    Jiang, Hao; Ouyang, Zheng; Zeng, Jianing; Yuan, Long; Zheng, Naiyu; Jemal, Mohammed; Arnold, Mark E

    2012-06-01

    Biological sample dilution is a rate-limiting step in bioanalytical sample preparation when the concentrations of samples are beyond standard curve ranges, especially when multiple dilution factors are needed in an analytical run. We have developed and validated a Microsoft Excel-based robotic sample preparation program (RSPP) that automatically transforms Watson worklist sample information (identification, sequence and dilution factor) to comma-separated value (CSV) files. The Freedom EVO liquid handler software imports and transforms the CSV files to executable worklists (.gwl files), allowing the robot to perform sample dilutions at variable dilution factors. The dynamic dilution range is 1- to 1000-fold and divided into three dilution steps: 1- to 10-, 11- to 100-, and 101- to 1000-fold. The whole process, including pipetting samples, diluting samples, and adding internal standard(s), is accomplished within 1 h for two racks of samples (96 samples/rack). This platform also supports online sample extraction (liquid-liquid extraction, solid-phase extraction, protein precipitation, etc.) using 96 multichannel arms. This fully automated and validated sample dilution and preparation process has been applied to several drug development programs. The results demonstrate that application of the RSPP for fully automated sample processing is efficient and rugged. The RSPP not only saved more than 50% of the time in sample pipetting and dilution but also reduced human errors. The generated bioanalytical data are accurate and precise; therefore, this application can be used in regulated bioanalysis.

  10. Evaluation of neon focused ion beam milling for TEM sample preparation.

    PubMed

    Pekin, T C; Allen, F I; Minor, A M

    2016-10-01

    Gallium-based focused ion beams generated from liquid-metal sources are widely used in micromachining and sample preparation for transmission electron microscopy, with well-known drawbacks such as sample damage and contamination. In this work, an alternative (neon) focused ion beam generated by a gas field-ionization source is evaluated for the preparation of electron-transparent specimens. To do so, electron-transparent sections of Si and an Al alloy are prepared with both Ga and Ne ion beams for direct comparison. Diffraction-contrast imaging and energy dispersive x-ray spectroscopy are used to evaluate the relative damage induced by the two beams, and cross-sections of milled trenches are examined to compare the implantation depth with theoretical predictions from Monte Carlo simulations. Our results show that for the beam voltages and materials systems investigated, Ne ion beam milling does not significantly reduce the focused ion beam induced artefacts. However, the Ne ion beam does enable more precise milling and may be of interest in cases where Ga contamination cannot be tolerated.

  11. Enhancing sample preparation capabilities for accelerator mass spectrometry radiocarbon and radiocalcium studies

    SciTech Connect

    Taylor, R E

    1991-08-20

    With support provided by the LLNL Accelerator Mass Spectrometry Laboratory, the UCR Radiocarbon Laboratory continued its studies involving sample pretreatment and target preparation for both AMS radiocarbon ({sup 14}C) and radiocalcium ({sup 41}Ca) involving applications to archaeologically -- and paleoanthropologically- related samples. With regard to AMS {sup 14}C-related studies, we have extended the development of a series of procedures which have, as their initial goal, the capability to combust several hundred microgram amounts of a chemically-pretreated organic sample and convert the resultant CO{sub 2} to graphitic carbon which will consistently yield relatively high {sup 13}C{sup {minus}} ion currents and blanks which will yield, on a consistent basis, {sup 14}C count rates at or below 0.20% modern, giving an 2 sigma age limit of >50,000 yr BP.

  12. Rapid determination of ethylene oxide with fiber-packed sample preparation needle.

    PubMed

    Ueta, Ikuo; Saito, Yoshihiro; Ghani, Nadia Binti Abdul; Ogawa, Mitsuhiro; Yogo, Kentaro; Abe, Akira; Shirai, Shingoro; Jinno, Kiyokatsu

    2009-04-03

    Fiber-packed sample preparation device was applied to the simultaneous derivatization/preconcentration of ethylene oxide (EO) in air samples. The polymer-coated filaments were packed longitudinally into the needle, and hydrogen bromide (HBr) was loaded onto the filaments in the preconditioning process. Simultaneous derivatization with HBr in the needle was made during the sampling process of the gaseous EO, and the corresponding derivatized analyte, 2-bromoethanol, was desorbed by passing a small amount of methanol through the extraction needle in the heated gas chromatograph (GC) injector. The basic extraction/desorption parameters for EO have been evaluated. The limit of detection (LOD), limit of quantification (LOQ) and the relative standard deviation (RSD) of run-to-run repeatability were 1.8 ng/L, 5.4 ng/L and less than 4%, respectively, with an extraction time of about 10 min. Satisfactory storage performance for three days at room temperature was also confirmed.

  13. Archive of Samples for Long-Term Preservation of RNA and Other Nucleic Acids

    DTIC Science & Technology

    2016-05-15

    nucleic acids W81XWH-12-C-0030 Sb. GRANT NUMBER Sc. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) Sd. PROJECT NUMBER Gerald Dodson Se. TASK NUMBER Sf. WORK...the development of a scalable all ambient temperature biological sample workflow preserving nucleic acids for molecular analysis. This ambient...achieved the final Phase II milestone of processing 500 samples n a week. 1S. SUBJECT TERMS Nucleic Acid Preservation , Cost Reduction , Ambient

  14. Extraction Chromatographic Methods in the Sample Preparation Sequence for Thermal Ionization Mass Spectrometric Analysis of Plutonium Isotopes

    SciTech Connect

    Grate, Jay W.; O'Hara, Matthew J.; Farawila, Anne F.; Douglas, Matthew; Haney, Morgan M.; Peterson, Steve L.; Maiti, Tapas C.; Aardahl, Christopher L.

    2011-10-17

    A sample preparation sequence for actinide isotopic analysis by TIMS is described that includes column-based extraction chromatography as the first separation step, followed by anion exchange column separations. The sequence is designed to include a wet ashing step after the extraction chromatography to prevent any leached extractant or oxalic acid eluent reagents from interfering with subsequent separations, source preparation, or TIMS ionization. TEVA-resin and DGA-resin materials, containing extractants that consist only of C, N, O, and H atoms, were investigated for isolation of plutonium. Radiotracer level studies confirmed expected high yields from column-based separation procedures. Femtogram-level studies were carried out with TIMS detection, using multiple isotopic spikes through the separation sequence. Pu recoveries were 87% and 86% for TEVA- and DGA-resins separations respectively. The Pu recoveries from 400 {mu}L anion-exchange column separations were 89% and 93% for trial sequences incorporating TEVA and DGA-resin. Thus, a prior extraction chromatography step in the sequence did not interfere with the subsequent anion exchange separation when a simple wet ash step was carried out in between these column separations. The average measurement efficiency, for Pu, encompassing the chemical separation recoveries and the TIMS ionization efficiency, was 2.73 {+-} 0.77% (2-sigma) for the DGA-resin trials and 2.67 {+-} 0.54% for the TEVA-resin trials, compared to 3.41% and 2.37% (average 2.89%) for two spikes in the experimental set. These compare with an average measurement efficiency of 2.78 {+-} 1.70%, n = 33 from process benchmark analyses using Pu spikes processed through a sequence of oxalate precipitation, wet ash, iron hydroxide precipitation, and anion exchange column separations. We conclude that extraction chromatography can be a viable separation procedure as part of a multistep sequence for TIMS sample preparation.

  15. [Quantitative analysis of seven phenolic acids in eight Yinqiao Jiedu serial preparations by quantitative analysis of multi-components with single-marker].

    PubMed

    Wang, Jun-jun; Zhang, Li; Guo, Qing; Kou, Jun-ping; Yu, Bo-yang; Gu, Dan-hua

    2015-04-01

    The study aims to develop a unified method to determine seven phenolic acids (neochlorogenic acid, chlorogenic acid, 4-caffeoylquinic acid, caffeic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) contained in honeysuckle flower that is the monarch drug of all the eight Yinqiao Jiedu serial preparations using quantitative analysis of multi-components by single-marker (QAMS). Firstly, chlorogenic acid was used as a reference to get the average relative correction factors (RCFs) of the other phenolic acids in ratios to the reference; columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions; and honeysuckle flower extract was used as the reference substance to fix the positions of chromatographic peaks. Secondly, the contents of seven phenolic acids in eight different Yinqiao Jiedu serial preparations samples were calculated based on the RCFs durability. Finally, the quantitative results were compared between QAMS and the external standard (ES) method. The results have showed that the durability of the achieved RCFs is good (RSD during 0.80% - 2.56%), and there are no differences between the quantitative results of QAMS and ES (the relative average deviation < 0.93%). So it can be successfully used to the quantitative control of honeysuckle flower principally prescribed in Yinqiao Jiedu serial preparations.

  16. Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

    PubMed Central

    Boardman, Anna K.; Campbell, Jennifer; Wirz, Holger; Sharon, Andre; Sauer-Budge, Alexis F.

    2015-01-01

    Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1–100 CFU/mL), traditionally requiring growth (blood culture) or amplification (e.g., PCR) for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components). Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms’ viability, giving a 30‑μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans) at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40–50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSA)to prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample. PMID:25675242

  17. Preparation and characterization of uniform particles of uric acid and its salts.

    PubMed

    Mohamed, Amr Ali; Matijević, Egon

    2013-02-15

    Uric acid, the major component in many kinds of kidney stones, as well as its sodium, ammonium, calcium, and barium salts were successfully prepared as uniform dispersions by precipitation in basic aqueous solutions. The effects of the reactant concentrations, pH, and the stabilizers were evaluated in detail. Except for the platelets of the pure acid, all prepared compounds appeared as needles or their aggregates. The electron micrographs showed that kidney stones consisted of such aggregates although less regular in size and morphology. All prepared urate salts had a 1:1 cation/uric acid ratio, regardless of the valence of the cation. The electrokinetic measurements showed all these particles to have negative ζ-potentials over the pH range 3-9. The precipitated salt particles were chemically and morphologically unstable at low pH values by decomposing into ill-defined aggregates of the pure uric acid.

  18. Flavonoids, cinnamic acids and coumarins from the different tissues and medicinal preparations of Taraxacum officinale.

    PubMed

    Williams, C A; Goldstone, F; Greenham, J

    1996-05-01

    Three flavonoid glycosides: luteolin 7-glucoside and two luteolin 7-diglucosides were isolated from dandelion flowers and leaves together with free luteolin and chrysoeriol in the flower tissue. The hydroxycinnamic acids, chicoric acid, monocaffeyltartaric acid and chlorogenic acid were found throughout the plant and the coumarins, cichoriin and aesculin were identified in the leaf extracts. This represents the first report of free chrysoeriol (luteolin 3'-methyl ether) in Taraxacum officinale agg. An earlier provisional identification of chicoric acid, chlorogenic acid, cichoriin and aesculin in a phenolic survey of the tribe Cichorieae is confirmed. Chicoric acid and the related monocaffeyltartaric acid were found to be the major phenolic constituents in flowers, roots, leaves and involucral bracts and also in the medicinal preparations tested.

  19. 14C sample preparation for AMS microdosing studies at Lund University using online combustion and septa-sealed vials

    NASA Astrophysics Data System (ADS)

    Sydoff, Marie; Stenström, Kristina

    2010-04-01

    The Department of Physics at Lund University is participating in a European Union project called EUMAPP (European Union Microdose AMS Partnership Programme), in which sample preparation and accelerator mass spectrometry (AMS) measurements of biological samples from microdosing studies have been made. This paper describes a simplified method of converting biological samples to solid graphite for 14C analysis with AMS. The method is based on online combustion of the samples, and reduction of CO 2 in septa-sealed vials. The septa-sealed vials and disposable materials are used to eliminate sample cross-contamination. Measurements of ANU and Ox I standards show deviations of 2% and 3%, respectively, relative to reference values. This level of accuracy is sufficient for biological samples from microdosing studies. Since the method has very few handling steps from sample to graphite, the risk of failure during the sample preparation process is minimized, making the method easy to use in routine preparation of samples.

  20. [Determination of glycyrrhizic acid in glycyrrhiza preparations with capillary electrophoresis and high performance liquid chromatography].

    PubMed

    Peng, J; Wang, F; Zhu, M

    1999-01-01

    High performance liquid chromatography (HPLC), capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography(MECC) have been used to determine glycyrrhizic acid in glycyrrhiza preparations. By HPLC, mobile phase was V(methanol):V(water):V(acetic acid) = 75:23.5:1.5. By CZE, experiment was performed with 15 kV power, 0.075 mm i.d. x 800 mm fused-silica capillary column and UV detector. Samples were injected into the capillary by electromigration injection for 20 s. Absorbance detection was at 254 nm. The running buffer was made up of 0.02 mol/L dipotassium hydrogenphosphate and borax (pH 9.0). By MECC, the running buffer was made up of 0.025 mol/L sodium dodecyl sulfate (SDS), 0.02 mol/L dipotassium hydrogenphosphate and borax (pH 9.0). Each new capillary was washed with 0.1 mol/L NaOH, deionized water and buffer, each for 3 min, before use. Comparison of the results of analysis with HPLC, CZE and MECC has been made. It was found that result of MECC was very close to that of HPLC. MECC has been satisfactorily applied to plant drug analysis.

  1. Arundo donax cane as a precursor for activated carbons preparation by phosphoric acid activation.

    PubMed

    Vernersson, T; Bonelli, P R; Cerrella, E G; Cukierman, A L

    2002-06-01

    Canes from Arundo donax, a herbaceous rapid-growing plant, were used as precursor for activated carbon preparation by phosphoric acid activation under a self-generated atmosphere. The influence of the carbonization temperature in the range 400-550 degrees C and of the weight ratio phosphoric acid to precursor (R = 1.5-2.5) on the developed porous structure of the resulting carbons was studied for 1 h of carbonization time. Surface properties of the activated carbons were dependent on a combined effect of the conditions employed. Carbons developed either with R = 1.5 over the range 400-500 degrees C, or with R = 2 at 500 degrees C exhibited surface areas of around 1100 m2/g, the latter conditions promoting a larger pore volume and enhanced mesoporous character. For both ratios, temperature above 500 degrees C led to reduction in porosity development. A similar effect was found for the highest ratio (R = 2.5) and 500 degrees C. The influence of carrying out the carbonization either for times shorter than 1 h or under flowing N2 was also examined at selected conditions (R = 2, 500 degrees C). Shorter times induced increase in the surface area (approximately 1300 m2/g), yielding carbons with smaller mean pore radius. Activated carbons obtained under flowing N2 possessed predominant microporous structures and larger ash contents than the samples derived in the self-generated atmosphere.

  2. Sample preparation for single virion atomic force microscopy and super-resolution fluorescence imaging.

    PubMed

    Hodges, Jeffery A; Saffarian, Saveez

    2014-01-02

    Immobilization of virions to glass surfaces is a critical step in single virion imaging. Here we present a technique adopted from single molecule imaging assays which allows adhesion of single virions to glass surfaces with specificity. This preparation is based on grafting the surface of the glass with a mixture of PLL-g-PEG and PLL-g-PEG-Biotin, adding a layer of avidin, and finally creating virion anchors through attachment of biotinylated virus specific antibodies. We have applied this technique across a range of experiments including atomic force microscopy (AFM) and super-resolution fluorescence imaging. This sample preparation method results in a control adhesion of the virions to the surface.

  3. Biological sample preparation and {sup 41}Ca AMS measurement at LLNL

    SciTech Connect

    Freeman, S.P.H.T.; Southon, J.R.; Bench, G.S.; McAninch, J.E.; Serfass, R.E.; Fang, Y.; King, J.C.; Woodhouse, L.R.

    1994-10-10

    Calcium metabolism in biology may be better understood by the use of {sup 41}Ca labels, although detection by accelerator mass spectrometry (AMS) is required. Methodologies for preparation of urine samples and subsequent AMS measurement were investigated. Novel attempts at preparing CaH{sub 2} were unsuccessful, but CaF{sub 2} of sufficient purity could be produced by precipitation of calcium from urine as oxalate, followed by separation of calcium by cation exchange chromatography and washing the CaF{sub 2} precipitate. The presence of some remaining impurities could be compensated for by selecting the appropriate accelerated ion charge state for AMS. The use of projectile x rays for isobar discrimination was explored as an alternative to the conventional dE/dx device.

  4. microPREP: a new laser tool for high-volume sample preparation

    NASA Astrophysics Data System (ADS)

    Wagner, Uwe; Petsch, Tino; Krause, Michael; Höche, Thomas

    2016-03-01

    Over the past fifty year, lasers have perpetuated to find new, often groundbreaking applications in science and technology. The most important features of lasers are that photons are inherently free of elemental contamination, extremely high energy densities can be focused in very small areas and the laser beam can be precisely positioned using deflection mirrors. By reducing pulse lengths from a few nanoseconds down to the picosecond or femtosecond range, material's ablation is becoming increasingly "athermal", i.e. structure damage by local heating is reduced to well below a few microns. In view of these outstanding characteristics of lasers as tools for micromachining, it is very surprising that sample preparation for microstructure diagnostics so far hasn't made use of laser technology. microPREPTM, the all-new, patented laser-micromachining tool developed by 3D-Micromac is the first instrument to make fast, clean, and efficient laser ablation available for the preparation of samples for microstructure diagnostics. Exemplified for a sample to be investigated by transmission electron microscopy (TEM) and following a three-stage approach, a supporting basic structure is cut from the feedstock first. Second, the supported structure is thinned down to a few micron of residual thickness and third, the supported and thinned structure is polished using an ion broad beam. Illustrated by numerous examples, it is shown that this technology is ready to be applied on different areas of microstructure diagnostics and has very high potential for failure diagnostics.

  5. A sample preparation method for recovering suppressed analyte ions in MALDI TOF MS.

    PubMed

    Lou, Xianwen; de Waal, Bas F M; Milroy, Lech-Gustav; van Dongen, Joost L J

    2015-05-01

    In matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF MS), analyte signals can be substantially suppressed by other compounds in the sample. In this technical note, we describe a modified thin-layer sample preparation method that significantly reduces the analyte suppression effect (ASE). In our method, analytes are deposited on top of the surface of matrix preloaded on the MALDI plate. To prevent embedding of analyte into the matrix crystals, the sample solution were prepared without matrix and efforts were taken not to re-dissolve the preloaded matrix. The results with model mixtures of peptides, synthetic polymers and lipids show that detection of analyte ions, which were completely suppressed using the conventional dried-droplet method, could be effectively recovered by using our method. Our findings suggest that the incorporation of analytes in the matrix crystals has an important contributory effect on ASE. By reducing ASE, our method should be useful for the direct MALDI MS analysis of multicomponent mixtures.

  6. Natural abiotic formation of oxalic acid in soils: results from aromatic model compounds and soil samples.

    PubMed

    Studenroth, Sabine; Huber, Stefan G; Kotte, Karsten; Schöler, Heinz F

    2013-02-05

    Oxalic acid is the smallest dicarboxylic acid and plays an important role in soil processes (e.g., mineral weathering and metal detoxification in plants). We have first proven its abiotic formation in soils and investigated natural abiotic degradation processes based on the oxidation of soil organic matter, enhanced by Fe(3+) and H(2)O(2) as hydroxyl radical suppliers. Experiments with the model compound catechol and further hydroxylated benzenes were performed to examine a common degradation pathway and to presume a general formation mechanism of oxalic acid. Two soil samples were tested for the release of oxalic acid and the potential effects of various soil parameters on oxalic acid formation. Additionally, the soil samples were treated with different soil sterilization methods to prove the oxalic acid formation under abiotic soil conditions. Different series of model experiments were conducted to determine a range of factors including Fe(3+), H(2)O(2), reaction time, pH, and chloride concentration on oxalic acid formation. Under certain conditions, catechol is degraded up to 65.6% to oxalic acid referring to carbon. In serial experiments with two soil samples, oxalic acid was produced, and the obtained results are suggestive of an abiotic degradation process. In conclusion, Fenton-like conditions with low Fe(3+) concentrations and an excess of H(2)O(2) as well as acidic conditions were required for an optimal oxalic acid formation. The presence of chloride reduced oxalic acid formation.

  7. Correlating Mineralogy and Amino Acid Contents of Milligram-Scale Murchison Carbonaceous Chondrite Samples

    NASA Technical Reports Server (NTRS)

    Burton, Aaron, S.; Berger, Eve L.; Locke, Darren R.; Elsila, Jamie E.; Glavin, Daniel P.; Dworkin, Jason P.

    2015-01-01

    Amino acids, the building blocks of proteins, have been found to be indigenous in most of the carbonaceous chondrite groups. The abundances of amino acids, as well as their structural, enantiomeric and isotopic compositions differ significantly among meteorites of different groups and petrologic types. This suggests that there is a link between parent-body conditions, mineralogy and the synthesis and preservation of amino acids (and likely other organic molecules). However, elucidating specific causes for the observed differences in amino acid composition has proven extremely challenging because samples analyzed for amino acids are typically much larger ((is) approximately 100 mg powders) than the scale at which meteorite heterogeneity is observed (sub mm-scale differences, (is) approximately 1-mg or smaller samples). Thus, the effects of differences in mineralogy on amino acid abundances could not be easily discerned. Recent advances in the sensitivity of instrumentation have made possible the analysis of smaller samples for amino acids, enabling a new approach to investigate the link between mineralogical con-text and amino acid compositions/abundances in meteorites. Through coordinated mineral separation, mineral characterization and highly sensitive amino acid analyses, we have performed preliminary investigations into the relationship between meteorite mineralogy and amino acid composition. By linking amino acid data to mineralogy, we have started to identify amino acid-bearing mineral phases in different carbonaceous meteorites. The methodology and results of analyses performed on the Murchison meteorite are presented here.

  8. Simple, Expendable, 3D-Printed Microfluidic Systems for Sample Preparation of Petroleum.

    PubMed

    Kataoka, Érica M; Murer, Rui C; Santos, Jandyson M; Carvalho, Rogério M; Eberlin, Marcos N; Augusto, Fabio; Poppi, Ronei J; Gobbi, Angelo L; Hantao, Leandro W

    2017-03-21

    In this study, we introduce a simple protocol to manufacture disposable, 3D-printed microfluidic systems for sample preparation of petroleum. This platform is produced with a consumer-grade 3D-printer, using fused deposition modeling. Successful incorporation of solid-phase extraction (SPE) to microchip was ensured by facile 3D element integration using proposed approach. This 3D-printed μSPE device was applied to challenging matrices in oil and gas industry, such as crude oil and oil-brine emulsions. Case studies investigated important limitations of nonsilicon and nonglass microchips, namely, resistance to nonpolar solvents and conservation of sample integrity. Microfluidic features remained fully functional even after prolonged exposure to nonpolar solvents (20 min). Also, 3D-printed μSPE devices enabled fast emulsion breaking and solvent deasphalting of petroleum, yielding high recovery values (98%) without compromising maltene integrity. Such finding was ascertained by high-resolution molecular analyses using comprehensive two-dimensional gas chromatography and gas chromatography/mass spectrometry by monitoring important biomarker classes, such as C10 demethylated terpanes, ααα-steranes, and monoaromatic steroids. 3D-Printed chips enabled faster and reliable preparation of maltenes by exhibiting a 10-fold reduction in sample processing time, compared to the reference method. Furthermore, polar (oxygen-, nitrogen-, and sulfur-containing) analytes found in low-concentrations were analyzed by Fourier transform ion cyclotron resonance mass spectrometry. Analysis results demonstrated that accurate characterization may be accomplished for most classes of polar compounds, except for asphaltenes, which exhibited lower recoveries (82%) due to irreversible adsorption to sorbent phase. Therefore, 3D-printing is a compelling alternative to existing microfabrication solutions, as robust devices were easy to prepare and operate.

  9. Error baseline rates of five sample preparation methods used to characterize RNA virus populations

    PubMed Central

    Kugelman, Jeffrey R.; Wiley, Michael R.; Nagle, Elyse R.; Reyes, Daniel; Pfeffer, Brad P.; Kuhn, Jens H.; Sanchez-Lockhart, Mariano; Palacios, Gustavo F.

    2017-01-01

    Individual RNA viruses typically occur as populations of genomes that differ slightly from each other due to mutations introduced by the error-prone viral polymerase. Understanding the variability of RNA virus genome populations is critical for understanding virus evolution because individual mutant genomes may gain evolutionary selective advantages and give rise to dominant subpopulations, possibly even leading to the emergence of viruses resistant to medical countermeasures. Reverse transcription of virus genome populations followed by next-generation sequencing is the only available method to characterize variation for RNA viruses. However, both steps may lead to the introduction of artificial mutations, thereby skewing the data. To better understand how such errors are introduced during sample preparation, we determined and compared error baseline rates of five different sample preparation methods by analyzing in vitro transcribed Ebola virus RNA from an artificial plasmid-based system. These methods included: shotgun sequencing from plasmid DNA or in vitro transcribed RNA as a basic “no amplification” method, amplicon sequencing from the plasmid DNA or in vitro transcribed RNA as a “targeted” amplification method, sequence-independent single-primer amplification (SISPA) as a “random” amplification method, rolling circle reverse transcription sequencing (CirSeq) as an advanced “no amplification” method, and Illumina TruSeq RNA Access as a “targeted” enrichment method. The measured error frequencies indicate that RNA Access offers the best tradeoff between sensitivity and sample preparation error (1.4−5) of all compared methods. PMID:28182717

  10. Preserving the distribution of inorganic arsenic species in groundwater and acid mine drainage samples

    USGS Publications Warehouse

    Bednar, A.J.; Garbarino, J.R.; Ranville, J.F.; Wildeman, T.R.

    2002-01-01

    The distribution of inorganic arsenic species must be preserved in the field to eliminate changes caused by metal oxyhydroxide precipitation, photochemical oxidation, and redox reactions. Arsenic species sorb to iron and manganese oxyhydroxide precipitates, and arsenite can be oxidized to arsenate by photolytically produced free radicals in many sample matrices. Several preservatives were evaluated to minimize metal oxyhydroxide precipitation, such as inorganic acids and ethylenediaminetetraacetic acid (EDTA). EDTA was found to work best for all sample matrices tested. Storing samples in opaque polyethylene bottles eliminated the effects of photochemical reactions. The preservation technique was tested on 71 groundwater and six acid mine drainage samples. Concentrations in groundwater samples reached 720 ??g-As/L for arsenite and 1080 ??g-As/L for arsenate, and acid mine drainage samples reached 13 000 ??g-As/L for arsenite and 3700 ??g-As/L for arsenate. The arsenic species distribution in the samples ranged from 0 to 90% arsenite. The stability of the preservation technique was established by comparing laboratory arsenic speciation results for samples preserved in the field to results for subsamples speciated onsite. Statistical analyses indicated that the difference between arsenite and arsenate concentrations for samples preserved with EDTA in opaque bottles and field speciation results were analytically insignificant. The percentage change in arsenite:arsenate ratios for a preserved acid mine drainage sample and groundwater sample during a 3-month period was -5 and +3%, respectively.

  11. Workshop on Mars 2001: Integrated Science in Preparation for Sample Return and Human Exploration

    NASA Technical Reports Server (NTRS)

    Marshall, John (Editor); Weitz, Cathy (Editor)

    1999-01-01

    The Workshop on Mars 2001: Integrated Science in Preparation for Sample Return and Human Exploration was held on October 2-4, 1999, at the Lunar and Planetary Institute in Houston, Texas. The workshop was sponsored by the Lunar and Planetary Institute, the Mars Program Office of the Jet Propulsion Laboratory, and the National Aeronautics and Space Administration. The three-day meeting was attended by 133 scientists whose purpose was to share results from recent missions, to share plans for the 2001 mission, and to come to an agreement on a landing site for this mission.

  12. [PREPARATIONS OF PAMIDRONOVIC ACID IN COMPLEX TREATMENT ON OSTEOGENESIS IMPERFECTA].

    PubMed

    Zyma, A M; Guk, Yu M; Magomedov, O M; Gayko, O G; Kincha-Polishchuk, T A

    2015-07-01

    Modern view of drug therapy in the complex treatment of orthopedic manifestations of osteogenesis imperfecta (OI) was submitted. Developed and tested system of drug correction of structural and functional state of bone tissue (BT) using drugs pamidronovic acid, depending on osteoporosis severity and type of disease. Such therapy is appropriate to apply both independently and in conjunction with surgery to correct deformations of long bones of the lower extremities. Effectiveness and feasibility of the proposed methods of drug therapy was proved, most patients resume features walking and support.

  13. Preparation and characterization of biomass carbon-based solid acid catalyst for the esterification of oleic acid with methanol.

    PubMed

    Liu, Tiantian; Li, Zhilong; Li, Wei; Shi, Congjiao; Wang, Yun

    2013-04-01

    A solid acid catalyst, prepared by sulfonating carbonized corn straw, was proved to be an efficient and environmental benign catalyst for the esterification of oleic acid and methanol. Various synthetic parameters, such as carbonization temperature and time were systematically examined. It was found that the catalyst exhibited the highest acid density of 2.64 mmol/g by NaOH titration. A quantitative yield (98%) of ester was achieved, using the most active sulfonated catalyst at 333 K with a 7 wt.% catalyst/oleic acid ratio for 4h, at a 7:1 M ratio of methanol/oleic acid, while the commercial available Amberlyst-15 only gave 85% yield under the same reaction condition.

  14. Selenium speciation in acidic environmental samples: application to acid rain-soil interaction at Mount Etna volcano.

    PubMed

    Floor, Geerke H; Iglesías, Mònica; Román-Ross, Gabriela; Corvini, Philippe F X; Lenz, Markus

    2011-09-01

    Speciation plays a crucial role in elemental mobility. However, trace level selenium (Se) speciation analyses in aqueous samples from acidic environments are hampered due to adsorption of the analytes (i.e. selenate, selenite) on precipitates. Such solid phases can form during pH adaptation up till now necessary for chromatographic separation. Thermodynamic calculations in this study predicted that a pH<4 is needed to prevent precipitation of Al and Fe phases. Therefore, a speciation method with a low pH eluent that matches the natural sample pH of acid rain-soil interaction samples from Etna volcano was developed. With a mobile phase containing 20mM ammonium citrate at pH 3, selenate and selenite could be separated in different acidic media (spiked water, rain, soil leachates) in <10 min with a LOQ of 0.2 μg L(-1) using (78)Se for detection. Applying this speciation analysis to study acid rain-soil interaction using synthetic rain based on H(2)SO(4) and soil samples collected at the flanks of Etna volcano demonstrated the dominance of selenate over selenite in leachates from samples collected close to the volcanic craters. This suggests that competitive behavior with sulfate present in acid rain might be a key factor in Se mobilization. The developed speciation method can significantly contribute to understand Se cycling in acidic, Al/Fe rich environments.

  15. Background and Artifacts Generated by the by the Sample Preparation Experiment on SAM

    NASA Astrophysics Data System (ADS)

    Belmahdi, Imene; Buch, Arnaud; Szopa, Cyril; Freissinet, Caroline; Glavin, Daniel; Coll, Patrice; Cabane, Michel; Millan, Maeva; Eigenbrode, Jennifer; Navarro-Gonzalez, Rafael; Stern, Jennifer; Coscia, David; Bonnet, Jean-Yves; Teinturier, Samuel; Morisson, Marietta; Stambouli, Moncef; Dequaire, Tristan; Mahaffy, Paul

    2016-04-01

    Sample Analysis at Mars (SAM) is one of the instruments of the Mars Science Laboratory mission. Three analytical devices composed the SAM experiment: the Tunable Laser Spectrometer (TLS), the Gas Chromatography (GC) and the Mass Spectrometer (MS). To adapt the nature of a sample to the analytical devices used, a sample preparation and gas processing system implemented with (a) a pyrolysis system, (b) wet chemistry: MTBSTFA and TMAH (c) the hydrocarbon trap (silica beads, Tenax® TA and Carbosieve G) and the injection trap (Tenax® GR composed of Tenax® TA and 30% of graphite) are employed to concentrate volatiles released from the sample prior to GC-MS analysis. Our study investigates several propositions for chlorinated hydrocarbon formation detected in the SAM background by looking for: (a) all products coming from the interaction of Tenax® and perchlorates present on Mars, (b) also between some soil sample and perchlorates and (c) sources of chlorinated hydrocarbon precursors. Here we report on the detection of chlorohydrocarbon compounds and their potential origin.

  16. Integrated process for preparing a carboxylic acid from an alkane

    SciTech Connect

    Benderly, Abraham; Chadda, Nitin; Sevon, Douglass

    2011-12-20

    The present invention relates to an integrated process for producing unsaturated carboxylic acids from the corresponding C.sub.2-C.sub.4 alkane. The process begins with performance of thermally integrated dehydrogenation reactions which convert a C.sub.2-C.sub.4 alkane to its corresponding C.sub.2-C.sub.4 alkene, and which involve exothermically converting a portion of an alkane to its corresponding alkene by oxidative dehydrogenation in an exothermic reaction zone, in the presence of oxygen and a suitable catalyst, and then feeding the products of the exothermic reaction zone to an endothermic reaction zone wherein at least a portion of the remaining unconverted alkane is endothermically dehydrogenated to form an additional quantity of the same corresponding alkene, in the presence of carbon dioxide and an other suitable catalyst. The alkene products of the thermally integrated dehydrogenation reactions are then provided to a catalytic vapor phase partial oxidation process for conversion of the alkene to the corresponding unsaturated carboxylic acid or nitrile. Unreacted alkene and carbon dioxide are recovered from the oxidation product stream and recycled back to the thermally integrated dehydrogenation reactions.

  17. Evaluation of sample preparation methods for water activity determination in jerky and kippered beef: a research note.

    PubMed

    Harper, N M; Getty, K J K; Boyle, E A E

    2010-10-01

    Commercially available packaged whole muscle beef jerky, chopped and formed beef jerky, and kippered beef steak were obtained from retail stores to determine the effect of two sample preparation methods on water activity (a(w)). Intact samples were prepared by cutting product into a hexagonal shape with a 3.2 cm diameter. Diced samples were prepared by cutting the product into 0.4 x 0.4 cm squares. Whole muscle jerky a(w) was higher (0.016 units; P<0.001) using the intact method compared to the diced method. There was no difference (P>0.05) in a(w) levels between the two preparation methods for chopped and formed jerky or kippered beef steak. An intact sample preparation method is recommended for a(w) determination of whole muscle jerky to obtain a more conservative value, especially if a(w) is near the margin of safety.

  18. Oxidative cleavage with hydrogen peroxide: preparation of polycarboxylic acids from cyclic olefins.

    PubMed

    Fujitani, Kango; Mizutani, Toshihiro; Oida, Tatsuo; Kawase, Tokuzo

    2009-01-01

    Oxidative cleavage of carbon-carbon double bonds of cyclic olefins with hydrogen peroxide in the presence of heteropolyacids has been investigated as a clean and environmentally friendly preparation of polycarboxylic acids. In the presence of 12-tungstophospholic acid (H(3)PW(12)O(40)), adipic acid was obtained in 95% yield from cyclohexene in lipophilic phase and hydrogen peroxide in aqueous phase. In addition, 1,2,3,4-butanetetracarboxylic acid was also obtained in 87% yield from 1,2,3,6-tetrahydrophtharic acid anhydride, while endic acid anhydride did not afford corresponding 2,3,6-cyclopentanetetracarboxylic acid but only lactone compound was obtained. In this oxidation process, oxidative cleavage of carbon-carbon double bonds would proceed as the sequential reactions in which the rate determining step is oxidative cleavage of vicinal-diol compounds.

  19. Preparation of low-molecular-weight hyaluronic acid by ozone treatment.

    PubMed

    Wu, Yue

    2012-06-20

    Recently, low-molecular-weight hyaluronic acid has been reported to have novel features, such as free radical scavenging activities, antioxidant activities, promotion of excisional wound healing, etc. In the present work, degradation of native hyaluronic acid by ozone treatment was performed for preparation of low-molecular-weight hyaluronic acid. The molecular weight of native hyaluronic acid was reduced from 1535 to 87 kDa for 120 min at 40°C. The rate of reduction of molecular weight was 94.33%. The FT-IR, 13C NMR, and UV-vis spectra suggested that there was no obvious modification of chemical structure of low-molecular-weight hyaluronic acid. The use of degradation of native hyaluronic acid by ozone treatment can be a useful alternative for production of low-molecular-weight hyaluronic acid.

  20. Sample preparation and separation techniques for bioanalysis of morphine and related substances.

    PubMed

    Hansen, Steen Honoré

    2009-03-01

    In present time the use or misuse of morphine and its derivatives are monitored by assaying the presence of the drug and its metabolites in biofluids. In the present review, focus is placed on the sample preparation and on the separation techniques used in the current best practices of bioanalysis of morphine and its major metabolites. However, as methods for testing the misuse of heroin, a morphine derivative, often involve bioanalytical methods that cover a number of other illicit drug substances, such methods are also included in the review. Furthermore, the review also includes bioanalysis in a broader perspective as analysis of plant materials, cell cultures and environmental samples. The review is not intended to cover all publications that include bioanalysis of morphine but is more to be considered a view into the current best practices of bioanalysis of morphine, its metabolites and other related substances.

  1. Sample preparation for the determination of steroids (corticoids and anabolics) in feed using LC.

    PubMed

    Muñiz-Valencia, Roberto; Ceballos-Magaña, Silvia G; Gonzalo-Lumbreras, Raquel; Santos-Montes, Ana; Izquierdo-Hornillos, Roberto C

    2008-07-01

    An improved sample preparation procedure for the determination of 17 steroids (corticoids (CC) and androgenic anabolic steroids (AAS)), used potentially as growth promoters, in feed samples has been developed. This procedure is based on two reported LC-UV methods. The improved procedure includes a leaching process using ACN, saponification, and SPE using polymeric cartridges. The proposed method was validated according to the EU criteria established for quantitative screening methods in PFS. The extraction efficiencies, decision limits (CCalpha) and detection capabilities (CCbeta), for these compounds were in the ranges of 82-100%, 19-40, and 24-53 microg/kg, respectively. The repeatability and the within-laboratory reproducibility at 1.0, 1.5, and 2.0 CCbeta levels were smaller than 10%. Accuracy was in the 97-101% range. The robustness was evaluated using the Youden robustness test. This method was applied to the analysis of steroids in different kinds of FS with satisfactory results.

  2. Nucleic acid isolation from ecological samples--fungal associations, mycorrhizae.

    PubMed

    Koide, Roger T

    2005-01-01

    Mycorrhizal fungi are among the most common symbioses found in terrestrial ecosystems, both natural and managed. They are important for many reasons, but most notably because of their positive effects on plant growth, which are mediated by their uptake of nutrients from the soil and transport of these to the roots. Moreover, many edible fungi are mycorrhizal. The study of mycorrhizal fungi has been hampered by the inability to identify species and individuals in the soil. This has been greatly aided by DNA-based methods, which first require the extraction of DNA. Herein, I discuss some general concerns that must be considered when extracting and purifying DNA from ecological samples and offer specific methods for soil, mycorrhizal roots, and fruiting bodies. These methods are rapid, safe, effective, relatively inexpensive, and convenient because they are based on commercially available kits.

  3. Purifying Nucleic Acids from Samples of Extremely Low Biomass

    NASA Technical Reports Server (NTRS)

    La Duc, Myron; Osman, Shariff; Venkateswaran, Kasthuri

    2008-01-01

    A new method is able to circumvent the bias to which one commercial DNA extraction method falls prey with regard to the lysing of certain types of microbial cells, resulting in a truncated spectrum of microbial diversity. By prefacing the protocol with glass-bead-beating agitation (mechanically lysing a much more encompassing array of cell types and spores), the resulting microbial diversity detection is greatly enhanced. In preliminary studies, a commercially available automated DNA extraction method is effective at delivering total DNA yield, but only the non-hardy members of the bacterial bisque were represented in clone libraries, suggesting that this method was ineffective at lysing the hardier cell types. To circumvent such a bias in cells, yet another extraction method was devised. In this technique, samples are first subjected to a stringent bead-beating step, and then are processed via standard protocols. Prior to being loaded into extraction vials, samples are placed in micro-centrifuge bead tubes containing 50 micro-L of commercially produced lysis solution. After inverting several times, tubes are agitated at maximum speed for two minutes. Following agitation, tubes are centrifuged at 10,000 x g for one minute. At this time, the aqueous volumes are removed from the bead tubes and are loaded into extraction vials to be further processed via extraction regime. The new method couples two independent methodologies in such as way as to yield the highest concentration of PCR-amplifiable DNA with consistent and reproducible results and with the most accurate and encompassing report of species richness.

  4. Recent developments on field gas extraction and sample preparation methods for radiokrypton dating of groundwater

    NASA Astrophysics Data System (ADS)

    Yokochi, Reika

    2016-09-01

    Current and foreseen population growths will lead to an increased demand in freshwater, large quantities of which is stored as groundwater. The ventilation age is crucial to the assessment of groundwater resources, complementing the hydrological model approach based on hydrogeological parameters. Ultra-trace radioactive isotopes of Kr (81 Kr and 85 Kr) possess the ideal physical and chemical properties for groundwater dating. The recent advent of atom trap trace analyses (ATTA) has enabled determination of ultra-trace noble gas radioisotope abundances using 5-10 μ L of pure Kr. Anticipated developments will enable ATTA to analyze radiokrypton isotope abundances at high sample throughput, which necessitates simple and efficient sample preparation techniques that are adaptable to various sample chemistries. Recent developments of field gas extraction devices and simple and rapid Kr separation method at the University of Chicago are presented herein. Two field gas extraction devices optimized for different sampling conditions were recently designed and constructed, aiming at operational simplicity and portability. A newly developed Kr purification system enriches Kr by flowing a sample gas through a moderately cooled (138 K) activated charcoal column, followed by a gentle fractionating desorption. This simple process uses a single adsorbent and separates 99% of the bulk atmospheric gases from Kr without significant loss. The subsequent two stages of gas chromatographic separation and a hot Ti sponge getter further purify the Kr-enriched gas. Abundant CH4 necessitates multiple passages through one of the gas chromatographic separation columns. The presented Kr separation system has a demonstrated capability of extracting Kr with > 90% yield and 99% purity within 75 min from 1.2 to 26.8 L STP of atmospheric air with various concentrations of CH4. The apparatuses have successfully been deployed for sampling in the field and purification of groundwater samples.

  5. Preparation of 4-amino-2,4-dioxobutanoic acid

    DOEpatents

    Unkefer, Pat J.; Martinez, Rodolfo A.; Glass, David R.

    2016-03-22

    A process for synthesizing 4-amino-2,4-dioxobutanoate involves reacting a dialkyl oxalate with an alkoxide in ethanol to form a reaction mixture, and afterward adding an alkyl cyano acetate to the reaction mixture and allowing a reaction to proceed under conditions suitable to form a first reaction product of the formula diethyl 2-cyano-3-hydroxy-butenedioate, and then isolating the diethyl 2-cyano-3-hydroxy-butenedioate, and afterward reacting the diethyl-2-cyano-3-hydroxy-butenedioate with an aqueous hydroxide under conditions suitable to form 4-amino-2,4-dioxobutanoate. The 4-amino-2,4-dioxobutanoate may be acidified into 4-amino-2,4-dioxobutanoic acid.

  6. Review of online coupling of sample preparation techniques with liquid chromatography.

    PubMed

    Pan, Jialiang; Zhang, Chengjiang; Zhang, Zhuomin; Li, Gongke

    2014-03-07

    Sample preparation is still considered as the bottleneck of the whole analytical procedure, and efforts has been conducted towards the automation, improvement of sensitivity and accuracy, and low comsuption of organic solvents. Development of online sample preparation techniques (SP) coupled with liquid chromatography (LC) is a promising way to achieve these goals, which has attracted great attention. This article reviews the recent advances on the online SP-LC techniques. Various online SP techniques have been described and summarized, including solid-phase-based extraction, liquid-phase-based extraction assisted with membrane, microwave assisted extraction, ultrasonic assisted extraction, accelerated solvent extraction and supercritical fluids extraction. Specially, the coupling approaches of online SP-LC systems and the corresponding interfaces have been discussed and reviewed in detail, such as online injector, autosampler combined with transport unit, desorption chamber and column switching. Typical applications of the online SP-LC techniques have been summarized. Then the problems and expected trends in this field are attempted to be discussed and proposed in order to encourage the further development of online SP-LC techniques.

  7. Preparation of an immunoaffinity column and its application in sample cleanup for methandrostenolone residues detection.

    PubMed

    Wang, Yun; Xu, Yan; Zhang, Xun; Wang, Enlan; Dong, Ying

    2011-07-15

    Methandrostenolone (MA) is a steroid used as veterinary medicine on stockbreeding to promote animal growth. The use of MA has been strictly regulated because of its harmful effect on consumers. This paper describes the production of polyclonal antibody (pAb) against MA, the preparation of immunoaffinity column (IAC) and its potential application to the selective extraction of MA residues from animal tissue and feed samples. The produced pAb exhibited good sensitivity to MA with an IC(50) value of 5.6 ng/mL. The cross-reactivity values of the antibody with MA structurally related compounds of testosterone propionate (TP) and trenbolone (TR) were lower than 0.6%. By coupling the produced antibody with CNBr-activated Sepharose 4B, an IAC was prepared. 2% methanol and 80% methanol were selected as loading and eluting solution by optimization. The maximum capacity of the column for MA was approximately 334 ng/mL gel. The average recovery of 20, 40 and 60 ng/mL MA standard solutions from IACs was 97.9% with the relative standard deviation (RSD) among columns of 6.7%. After 3 times of repeated usage, the column capacity and recovery rate still remained 82.0% and 92.6% respectively. The IACs were then challenged with MA-fortified animal tissue and feed samples, recoveries of MA were found to be in the range of 83.5-99.7%.

  8. Matrix compatible solid phase microextraction coating, a greener approach to sample preparation in vegetable matrices.

    PubMed

    Naccarato, Attilio; Pawliszyn, Janusz

    2016-09-01

    This work proposes the novel PDMS/DVB/PDMS fiber as a greener strategy for analysis by direct immersion solid phase microextraction (SPME) in vegetables. SPME is an established sample preparation approach that has not yet been adequately explored for food analysis in direct immersion mode due to the limitations of the available commercial coatings. The robustness and endurance of this new coating were investigated by direct immersion extractions in raw blended vegetables without any further sample preparation steps. The PDMS/DVB/PDMS coating exhibited superior features related to the capability of the external PDMS layer to protect the commercial coating, and showed improvements in terms of extraction capability and in the cleanability of the coating surface. In addition to having contributed to the recognition of the superior features of this new fiber concept before commercialization, the outcomes of this work serve to confirm advancements in the matrix compatibility of the PDMS-modified fiber, and open new prospects for the development of greener high-throughput analytical methods in food analysis using solid phase microextraction in the near future.

  9. Semiautomated Sample Preparation for Protein Stability and Formulation Screening via Buffer Exchange.

    PubMed

    Ying, William; Levons, Jaquan K; Carney, Andrea; Gandhi, Rajesh; Vydra, Vicky; Rubin, A Erik

    2016-06-01

    A novel semiautomated buffer exchange process workflow was developed to enable efficient early protein formulation screening. An antibody fragment protein, BMSdab, was used to demonstrate the workflow. The process afforded 60% to 80% cycle time and scientist time savings and significant material efficiencies. These efficiencies ultimately facilitated execution of this stability work earlier in the drug development process, allowing this tool to inform the developability of potential candidates for development from a formulation perspective. To overcome the key technical challenges, the protein solution was buffer-exchanged by centrifuge filtration into formulations for stability screening in a 96-well plate with an ultrafiltration membrane, leveraging automated liquid handling and acoustic volume measurements to allow several cycles of exchanges. The formulations were transferred into a vacuum manifold and sterile filtered into a rack holding 96 glass vials. The vials were sealed with a capmat of individual caps and placed in stability stations. Stability of the samples prepared by this process and by the standard process was demonstrated to be comparable. This process enabled screening a number of formulations of a protein at an early pharmaceutical development stage with a short sample preparation time.

  10. Structural, morphological and catalytic characterization of neutral Ag salt of 12-tungstophosphoric acid: Influence of preparation conditions

    NASA Astrophysics Data System (ADS)

    Holclajtner-Antunović, Ivanka; Bajuk-Bogdanović, Danica; Popa, Alexandru; Nedić Vasiljević, Bojana; Krstić, Jugoslav; Mentus, Slavko; Uskoković-Marković, Snežana

    2015-02-01

    The objective of this study is the structural and morphological characterization of the Ag3PW12O40 salts (AgWPA) of 12-tungstophosphoric acid (WPA) obtained under different preparation conditions and testing of their acid catalytic activity in dehydration of ethanol. The structure, morphology and physicochemical characteristics were determined by Fourier transform infrared (FT-IR) and Raman spectroscopy, X-ray diffraction (XRD), nitrogen physisorption at -196 °C, scanning electron microscopy (SEM) and differential thermal (DTA) and thermogravimetric analysis (TGA). It is shown that the preparation process has a significant influence on the morphological properties of the obtained materials which may be explained by the supposed mechanism of the formation of nanocrystallite‧s aggregates with more or less epitaxial connection. Neutral AgWPA obtained by filtration from supernatant forms porous aggregates of a symmetric dodecahedral shape, having average sizes about 2 μm. This sample shows higher specific area in comparison with the salt obtained by evaporation due to the higher micropore volume, while mesopore volumes are the same for both salts. Thus conversion of ethanol and selectivities of the main products, ethylene and diethyl ether, are almost the same and constant for both prepared salts, while their values are changed over the reaction time for the parent WPA acid.

  11. The minimum information required for a glycomics experiment (MIRAGE) project: sample preparation guidelines for reliable reporting of glycomics datasets.

    PubMed

    Struwe, Weston B; Agravat, Sanjay; Aoki-Kinoshita, Kiyoko F; Campbell, Matthew P; Costello, Catherine E; Dell, Anne; Ten Feizi; Haslam, Stuart M; Karlsson, Niclas G; Khoo, Kay-Hooi; Kolarich, Daniel; Liu, Yan; McBride, Ryan; Novotny, Milos V; Packer, Nicolle H; Paulson, James C; Rapp, Erdmann; Ranzinger, Rene; Rudd, Pauline M; Smith, David F; Tiemeyer, Michael; Wells, Lance; York, William S; Zaia, Joseph; Kettner, Carsten

    2016-09-01

    The minimum information required for a glycomics experiment (MIRAGE) project was established in 2011 to provide guidelines to aid in data reporting from all types of experiments in glycomics research including mass spectrometry (MS), liquid chromatography, glycan arrays, data handling and sample preparation. MIRAGE is a concerted effort of the wider glycomics community that considers the adaptation of reporting guidelines as an important step towards critical evaluation and dissemination of datasets as well as broadening of experimental techniques worldwide. The MIRAGE Commission published reporting guidelines for MS data and here we outline guidelines for sample preparation. The sample preparation guidelines include all aspects of sample generation, purification and modification from biological and/or synthetic carbohydrate material. The application of MIRAGE sample preparation guidelines will lead to improved recording of experimental protocols and reporting of understandable and reproducible glycomics datasets.

  12. RNA SAMPLE PREPARATION APPLIED TO GENE EXPRESSION PROFILING FOR THE HORSE BIOLOGICAL PASSPORT.

    PubMed

    Bailly-Chouriberry, Ludovic; Baudoin, Florent; Cormant, Florence; Glavieux, Yohan; Loup, Benoit; Garcia, Patrice; Popot, Marie-Agnès; Bonnaire, Yves

    2017-04-05

    The improvement of doping control is an on-going race. Techniques to fight against doping are usually based on the direct detection of drugs or their metabolites by analytical methods such as chromatography hyphenated to mass spectrometry after ad hoc sample preparation. Nowadays, omic methods constitute an attractive development and advances have been achieved particularly by application of molecular biology tools for detection of anabolic androgenic steroids (AAS), erythropoiesis-stimulating agent (ESA) or to control human growth hormone misuses. These interesting results across different animal species have suggested that modification of gene expression offers promising new methods of improving the window of detection of banned substances by targeting their effects on blood cell gene expression. In this context, the present study describes the possibility of using a modified version of the dedicated Human IVD (in vitro Diagnostics) PAXgene® Blood RNA Kit for horse gene expression analysis in blood collected on PAXgene® tubes applied to the Horse Biological Passport. The commercial kit was only approved for human blood samples and has required an optimization of specific technical requirements for equine blood samples. Improvements and recommendations were achieved for sample collection, storage and RNA extraction procedure. Following these developments, RNA yield and quality were demonstrated to be suitable for downstream gene expression analysis by qPCR techniques.

  13. Optimized sample preparation of endoscopic collected pancreatic fluid for SDS-PAGE analysis.

    PubMed

    Paulo, Joao A; Lee, Linda S; Wu, Bechien; Repas, Kathryn; Banks, Peter A; Conwell, Darwin L; Steen, Hanno

    2010-07-01

    The standardization of methods for human body fluid protein isolation is a critical initial step for proteomic analyses aimed to discover clinically relevant biomarkers. Several caveats have hindered pancreatic fluid proteomics, including the heterogeneity of samples and protein degradation. We aim to optimize sample handling of pancreatic fluid that has been collected using a safe and effective endoscopic collection method (endoscopic pancreatic function test). Using SDS-PAGE protein profiling, we investigate (i) precipitation techniques to maximize protein extraction, (ii) auto-digestion of pancreatic fluid following prolonged exposure to a range of temperatures, (iii) effects of multiple freeze-thaw cycles on protein stability, and (iv) the utility of protease inhibitors. Our experiments revealed that TCA precipitation resulted in the most efficient extraction of protein from pancreatic fluid of the eight methods we investigated. In addition, our data reveal that although auto-digestion of proteins is prevalent at 23 and 37 degrees C, incubation on ice significantly slows such degradation. Similarly, when the sample is maintained on ice, proteolysis is minimal during multiple freeze-thaw cycles. We have also determined the addition of protease inhibitors to be assay-dependent. Our optimized sample preparation strategy can be applied to future proteomic analyses of pancreatic fluid.

  14. Preparation of Magnetic Hollow Molecularly Imprinted Polymers for Detection of Triazines in Food Samples.

    PubMed

    Wang, Aixiang; Lu, Hongzhi; Xu, Shoufang

    2016-06-22

    Novel magnetic hollow molecularly imprinted polymers (M-H-MIPs) were proposed for highly selective recognition and fast enrichment of triazines in food samples. M-H-MIPs were prepared on the basis of multi-step swelling polymerization, followed by in situ growth of magnetic Fe3O4 nanoparticles on the surface of hollow molecularly imprinted polymers (H-MIPs). Transmission electron microscopy and scanning electron microscopy confirmed the successful immobilization of Fe3O4 nanoparticles on the surface of H-MIPs. M-H-MIPs could be separated simply using an external magnet. The binding adsorption results indicated that M-H-MIPs displayed high binding capacity and fast mass transfer property and class selective property for triazines. Langmuir isotherm and pseudo-second-order kinetic models fitted the best adsorption models for M-H-MIPs. M-H-MIPs were used to analyze atrazine, simazine, propazine, and terbuthylazine in corn, wheat, and soybean samples. Satisfactory recoveries were in the range of 80.62-101.69%, and relative standard deviation was lower than 5.2%. Limits of detection from 0.16 to 0.39 μg L(-1) were obtained. When the method was applied to test positive samples that were contaminated with triazines, the results agree well with those obtained from an accredited method. Thus, the M-H-MIP-based dispersive solid-phase extraction method proved to be a convenient and practical platform for detection of triazines in food samples.

  15. Polymerase chain reaction system using magnetic beads for analyzing a sample that includes nucleic acid

    DOEpatents

    Nasarabadi, Shanavaz [Livermore, CA

    2011-01-11

    A polymerase chain reaction system for analyzing a sample containing nucleic acid includes providing magnetic beads; providing a flow channel having a polymerase chain reaction chamber, a pre polymerase chain reaction magnet position adjacent the polymerase chain reaction chamber, and a post pre polymerase magnet position adjacent the polymerase chain reaction chamber. The nucleic acid is bound to the magnetic beads. The magnetic beads with the nucleic acid flow to the pre polymerase chain reaction magnet position in the flow channel. The magnetic beads and the nucleic acid are washed with ethanol. The nucleic acid in the polymerase chain reaction chamber is amplified. The magnetic beads and the nucleic acid are separated into a waste stream containing the magnetic beads and a post polymerase chain reaction mix containing the nucleic acid. The reaction mix containing the nucleic acid flows to an analysis unit in the channel for analysis.

  16. Citric acid traps to replace sulphuric acid in the ammonia diffusion of dilute water samples for 15N analysis.

    PubMed

    Schleppi, Patrick; Bucher-Wallin, Inga; Saurer, Matthias; Jäggi, Maya; Landolt, Werner

    2006-01-01

    The analysis of 15N in aqueous samples requires the concentration of dissolved nitrogen (N) into a small volume that can be analysed by mass spectrometry. This is conveniently achieved by the NH3 diffusion technique, where NH4+ is captured on small acidified filters enclosed in PTFE. NO3- can be analysed the same way by reducing it to NH4+ with Devarda's alloy. H2SO4 is commonly used for the acidification of the filters. During combustion, however, this acid leads to the production of SO2 and elemental sulphur, which both have detrimental effects on the mass spectrometer. We propose here to replace H2SO4 with citric acid because it is combusted completely to CO2 and H2O in the elemental analyser before entering the mass spectrometer. Citric acid was found to give comparable results in terms of N recovery and 15N values, both for NH4+ and for NO3- samples. Blank samples revealed that N contamination was slightly lower using citric instead of sulphuric acid as acidifier of the glass filters. NH4+ samples first concentrated over cation-exchange columns were strongly acidic and several methods were tested to raise the pH for the subsequent diffusion. These samples gave incomplete N recoveries, but this problem was independent of the acid used on the filters and of the final pH of the sample. Complete recovery was achieved only by increasing the volume of the eluate from the columns. Citric acid can thus generally be recommended instead of H2SO4 for ammonia diffusion.

  17. The search for and identification of amino acids, nucleobases and nucleosides in samples returned from Mars

    NASA Technical Reports Server (NTRS)

    Gehrke, Charles W.; Ponnamperuma, Cyril; Kuo, Kenneth C.; Stalling, David L.; Zumwalt, Robert W.

    1989-01-01

    An investigation of the returned Mars samples for biologically important organic compounds, with emphasis on amino acid, the puring and pyrimidine bases, and nucleosides is proposed. These studies would be conducted on subsurface samples obtained by drilling past the surface oxidizing layer with emphasis on samples containing the larges quantities of organic carbon as determined by the rover gas chromatographic mass spectrometer (GCMS). Extraction of these molecules from the returned samples will be performed using the hydrothermal extraction technique described by Cheng and Ponnamperuma. More rigorous extraction methods will be developed and evaluated. For analysis of the extract for free amino acids or amino acids present in a bound or peptidic form, aliquots will be analyzed by capillary GCMS both before and after hydrolysis with 6N hydrochloric acid. Establishment of the presence of amino acids would then lead to the next logical step which would be the use of chiral stationary gas chromatography phases to determine the enatiomeic composition of the amino acids present, and thus potentially establish their biotic or abiotic origin. Confirmational analyses for amino acids would include ion-exchange and reversed-phase liquid chromatographic analysis. For analyses of the returned Mars samples for nucleobases and nucleosides, affinity and reversed-phase liquid chromatography would be utilized. This technology coupled with scanning UV detection for identification, presents a powerful tool for nucleobase and nucleoside analysis. Mass spectrometric analysis of these compounds would confirm their presence in samples returned form Mars.

  18. Array capillary in-tube solid-phase microextraction: a rapid preparation technique for water samples.

    PubMed

    Yan, Xiaohui; Wu, Dapeng; Peng, Hong; Ding, Kun; Duan, Chunfeng; Guan, Yafeng

    2012-06-29

    A rapid sample preparation technique, namely array capillary in-tube solid-phase microextraction (ACIT-SPME) for direct extraction of organic pollutants from water samples, was developed and evaluated. The ACIT-SPME cartridge consisted of a bundle of glass capillary tubes of 0.5 mm I.D. × 30mm contained inside a quartz liner of 4 mm I.D. The high ratio of cross-section area of channel-to-wall allowed water sample flow through the cartridge just under gravity. Both the internal/external surfaces of the array capillary tubing were coated with extraction phase of 2-5 μm in thickness, which provided large extraction surface area up to 30 cm² for a cartridge containing 19 glass capillaries. The large surface area and thin extraction phase improved greatly both the mass transfer process of extraction and the thermo desorption process, leading to fast extraction and fast desorption. The extracted analytes were thermally desorbed in a homemade thermal desorption unit (TDU), which was coupled to a gas chromatograph equipped with a flame ionization detector for analysis. By using polydimethylsiloxane (PDMS) as the extraction phase and polycyclic aromatic hydrocarbons (PAHs) as the model analytes, the performance of the ACIT-SPME cartridge was systematically investigated. The extraction time was 2 min for 350 mL of water sample, and detection limits were between 0.8 and 1.7 ng/L with deviation of 2.8-9.7% RSD. Relative recoveries of analytes for real water samples were between 65.0% and 116%. The extraction time can even be further shortened to 10s for 250 mL sample by applying vacuum at the outlet of the cartridge, with detection limits of 2.2-5.3 ng/L and deviation of 4.0-12% RSD.

  19. Automation of sample preparation for mass cytometry barcoding in support of clinical research: protocol optimization.

    PubMed

    Nassar, Ala F; Wisnewski, Adam V; Raddassi, Khadir

    2017-03-01

    Analysis of multiplexed assays is highly important for clinical diagnostics and other analytical applications. Mass cytometry enables multi-dimensional, single-cell analysis of cell type and state. In mass cytometry, the rare earth metals used as reporters on antibodies allow determination of marker expression in individual cells. Barcode-based bioassays for CyTOF are able to encode and decode for different experimental conditions or samples within the same experiment, facilitating progress in producing straightforward and consistent results. Herein, an integrated protocol for automated sample preparation for barcoding used in conjunction with mass cytometry for clinical bioanalysis samples is described; we offer results of our work with barcoding protocol optimization. In addition, we present some points to be considered in order to minimize the variability of quantitative mass cytometry measurements. For example, we discuss the importance of having multiple populations during titration of the antibodies and effect of storage and shipping of labelled samples on the stability of staining for purposes of CyTOF analysis. Data quality is not affected when labelled samples are stored either frozen or at 4 °C and used within 10 days; we observed that cell loss is greater if cells are washed with deionized water prior to shipment or are shipped in lower concentration. Once the labelled samples for CyTOF are suspended in deionized water, the analysis should be performed expeditiously, preferably within the first hour. Damage can be minimized if the cells are resuspended in phosphate-buffered saline (PBS) rather than deionized water while waiting for data acquisition.

  20. Preparation of environmental samples for determination of sulfonylurea herbicides by solid-phase extraction using a polymeric sorbent.

    PubMed

    Young, M S

    1998-01-01

    An analytical procedure was developed for rapid determination of sulfonylurea herbicides in water or soil. Samples were prepared by solid-phase extraction using polymeric sorbents in cartridges and analyzed by liquid chromatography with photodiode array detection. Recoveries were consistent for all sample matrixes tested and were generally > 85%, except for tribenuron methyl (75%). Detection limits were < 100 ng/L for water samples.

  1. Studies on the oxidation of hexamethylbenzene 2: Preparation of dimethylpyromellitic acid

    NASA Technical Reports Server (NTRS)

    Chiba, K.; Tomura, S.

    1986-01-01

    Hexamethylbenzene (HMB) was difficult to be oxidized with an alkaline potassium permanganate solution, since HMB was insoluble in an aqueous alkaline solution. But, when HMB was warmed with 50% nitric acid for a short time, and then treated with aqueous potassium permanganate, the reaction occurred readily and dimethylpyromellitic acid was obtained. When HMB was warmed with 50% nitric acid for 1 to 2 minutes, a yellow material was produced, which was soluble in hot aqueous potassium hydroxide, though free from carboxylic acids. It contained a little amount of bis-(nitromethyl)prehnitene and several unknown compounds. Further, the heat stability of polyimide prepared by the reaction of tetramethyldimethylpyromellitate with 4,4 prime-diaminodiphenylmethane turned out to be nearly equal to that of polyimide prepared from tetramethylpyromellitate.

  2. Effect of the solvent and the sample preparation on the determination of triterpene compounds in two-phase olive-mill-waste samples.

    PubMed

    Fernández-Hernández, Antonia; Martinez, Antonio; Rivas, Francisco; García-Mesa, Jose A; Parra, Andres

    2015-05-06

    A simple and rapid extraction method has been employed to determine several value-added compounds, mainly triterpenes, in two-phase olive-mill-waste samples. The compounds were extracted with methanol or ethyl acetate, and the initial fresh samples were treated for classic techniques such as drying, drying and oil extraction, and drying and sifting of the olive stones. For the identification and quantitation of the compounds, an ultra performance liquid chromatography-mass spectrometry method was employed. The best results of the triterpenic compound content were achieved by extraction with methanol from the fresh sample for the oleanolic and ursolic acids, and erythrodiol and uvaol; and from the dried-extracted sample for the maslinic acid. Conversely, the best results for the linoleic acid content were reached by extraction with ethyl acetate from the dried-sifted sample. These are remarkable processes that make the solid wastes from the olive-oil industry reach a high added value.

  3. Preparation of a multi-hollow magnetic molecularly imprinted polymer for the selective enrichment of indolebutyric acid.

    PubMed

    Li, Shanshan; Yin, Chao; Ren, Shuiying; Yang, Tao; Wang, Jide; Feng, Shun

    2015-08-01

    A simple strategy was developed for the preparation of multi-hollow magnetic molecularly imprinted polymers by incorporating 3-indolebutyric acid and ferroferric oxide nanoparticles simultaneously into a poly(styrene-co-methacrylic acid) copolymer matrix. The as prepared absorbents were characterized using scanning electron microscopy, Fourier-transform infrared spectroscopy and mercury porosimetry. The adsorption isotherms of indolebutyric acid revealed that there are two types of affinity binding sites in the absorbents. The apparent maximum binding capacity and dissociation constant were 17.88 mg/g and 158.7 μg/mL for high-affinity binding sites and 9.310 mg/g and 35.04 μg/mL for low-affinity binding sites, respectively. The results testified that multi-hollow magnetic molecularly imprinted polymers possessed excellent recognition capacity and fast kinetic binding behavior to the objective molecules due to the high specific surface area as large as 511.3 m(2) /g. Recoveries of 75.5-86.8% were obtained for the indolebutyric acid spiked at three concentration levels in blank and pear samples.

  4. Simultaneous determination of acetylsalicylic and salicylic acids by first derivative spectrometry in pharmaceutical preparations

    NASA Astrophysics Data System (ADS)

    Rogić, Dunja

    1993-03-01

    A multicomponent first derivative UV spectrometric procedure for determination of acetylsalicylic acid (aspirin) and salicylic acid in the solution containing 1 % (w/v) of citric acid in some pharmaceutical preparations is presented. The method is based on the use of the first derivative minimum spectrometric measurements at 286 nm for aspirin and at 318 nm for salicylic acid. Four kinds of cmmercial Aspirin tablets were assayed without a long pretreatment of the pharmaceuticals from the tablet additives. Beer's law is obeyed from 13.62-68.1 μg ml -1 of aspirin and from 2.723-13.616 μg ml -1 of salicylic acid. Detection limits at the 0.05 level of significance were calculated to be 1.24 and 0.25 μg ml -1 with relative standard deviations of 1.09 % and 1.2 % of aspirin and salicylic acid, respectively.

  5. Dual-column cation-exchange chromatographic method for beta-aminoisobutyric acid and beta-alanine in biological samples.

    PubMed

    Kuo, K C; Cole, T F; Gehrke, C W; Waalkes, T P; Borek, E

    1978-08-01

    A rapid, automated chromatographic method has been developed for the quantitation of the nucleic acid catabolites beta-aminoisobutyric acid and beta-alanine in urine, serum, and other physiological fluids. The analyses were performed on a modified Beckman 121M amino acid analyzer with dual ion-exchange columns and the use of a single sodium citrate buffer (pH 4.38, 0.20 mol/liter). By carefully matching the elution pattern for the two ion-exchange columns and alternating use of these columns, analyses are completed every 40 min. The chromatography, regeneration, and equilibration of the two columns are precisely programmed, thus the detector sees only the elution of beta-aminoisobutyric acid and beta-alanine alternately from each column. Long-term precision and analytical recovery for the two metabolites in urine were 1.9 and 102%, and 3.3 and 101%, respectively. Their normal physiological values were determined in human serum and urine. Their excretion in the urine was also studied as a function of collection time, to validate a more convenient, less costly method of sampling. This study shows that randomly collected samples are acceptable when the concentration of the two metabolites are expressed in terms of creatinine excretion. In addition, the distribution of the free and conjugated forms of the two metabolites in urine and serum was studied. A preparative method was also developed for the quantitative isolation of beta-amino-isobutyric acid from urine samples. The alternating dual-column technique may be applied to any ion-exchange chromatographic method where many analyses must be performed. This method is currently used in our laboratories for measuring these beta-amino acids in urine and serum of patients with various types of cancers.

  6. Determination of Mycotoxins in Brown Rice Using QuEChERS Sample Preparation and UHPLC-MS-MS.

    PubMed

    Jettanajit, Adisorn; Nhujak, Thumnoon

    2016-01-01

    QuEChERS sample preparation was optimized and validated using solvent extraction with 10% (v/v) acetic acid-containing acetonitrile in the presence of four salts (anh. MgSO4, NaCl, sodium citrate tribasic dihydrate and sodium citrate dibasic sesquihydrate) and dispersive solid-phase extraction with mixed sorbents (octadecylsilane, primary and secondary amine and silica sorbents) for an ultra high performance liquid chromatography-tandem mass spectrometric determination of nine mycotoxins in brown rice: aflatoxins (AFB1, AFB2, AFG1 and AFG2), fumonisins (FB1 and FB2), deoxynivalenol, ochratoxin A and zearalenone (ZON). Our developed method allows for the determination of trace levels of mycotoxins with method detection limits in the range of 1.4-25 µg/kg, below the maximum limits of EU regulations, and with an acceptable accuracy and precision, and recoveries in the range of 81-101% with relative standard deviations of 5-19% over a mycotoxin concentration range of 5.0-1,000 µg/kg. Six out of fourteen real samples of brown rice were found to be contaminated with at least one of these mycotoxins, ranging from 2.49-5.41 µg/kg of FB1, 4.33 ± 0.04 µg/kg of FB2 and 6.10-14.88 µg/kg of ZON.

  7. Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR.

    PubMed

    Capote, Nieves; Bertolini, Edson; Olmos, Antonio; Vidal, Eduardo; Martínez, Maria Carmen; Cambra, Mariano

    2009-03-01

    Direct systems to process plant materials allowed high-throughput testing of Plum pox virus (PPV) by real-time reverse transcription (RT)-PCR without nucleic acids purification. Crude plant extracts were diluted in buffer or spotted on membranes to be used as templates. Alternatively, immobilized PPV targets were amplified from fresh sections of plant tissues printed or squashed onto the same supports, without extract preparation. Spot real-time RT-PCR was validated as a PPV diagnostic method in samples collected during the dormancy period and showed high sensitivity (93.6%), specificity (98.0%), and post-test probability (97.9%) towards sharka disease. In an analysis of 2919 Prunus samples by spot real-time RT-PCR and DASI-ELISA 90.8% of the results coincided, demonstrating high agreement (k = 0.77 +/- 0.01) between the two techniques. These results validate the use of immobilized PPV targets and spot real-time RT-PCR as screening method for largescale analyses.

  8. A 96-well screen filter plate for high-throughput biological sample preparation and LC-MS/MS analysis.

    PubMed

    Peng, Sean X; Cousineau, Martin; Juzwin, Stephen J; Ritchie, David M

    2006-01-01

    A novel 96-well screen filter plate (patent pending) has been invented to eliminate a time-consuming and labor-intensive step in preparation of in vivo study samples--to remove blood or plasma clots. These clots plug the pipet tips during a manual or automated sample-transfer step causing inaccurate pipetting or total pipetting failure. Traditionally, these blood and plasma clots are removed by picking them out manually one by one from each sample tube before any sample transfer can be made. This has significantly slowed the sample preparation process and has become a bottleneck for automated high-throughput sample preparation using robotic liquid handlers. Our novel screen filter plate was developed to solve this problem. The 96-well screen filter plate consists of 96 stainless steel wire-mesh screen tubes connected to the 96 openings of a top plate so that the screen filter plate can be readily inserted into a 96-well sample storage plate. Upon insertion, the blood and plasma clots are excluded from entering the screen tube while clear sample solutions flow freely into it. In this way, sample transfer can be easily completed by either manual or automated pipetting methods. In this report, three structurally diverse compounds were selected to evaluate and validate the use of the screen filter plate. The plasma samples of these compounds were transferred and processed in the presence and absence of the screen filter plate and then analyzed by LC-MS/MS methods. Our results showed a good agreement between the samples prepared with and without the screen filter plate, demonstrating the utility and efficiency of this novel device for preparation of blood and plasma samples. The device is simple, easy to use, and reusable. It can be employed for sample preparation of other biological fluids that contain floating particulates or aggregates.

  9. Efficient preparation of (R)-2-chloromandelic acid via a recycle process of resolution.

    PubMed

    Hu, Yu; Wu, Chang; Wu, Xue-Ying; Li, Shao-Lei; Sun, Xiao-Xia; Tang, Ze-Biao

    2015-03-01

    Efficient preparation of (R)-2-chloromandelic acid based on a recycle process of resolution is described. In the process, the desired was obtained by coordination-mediated resolution with D-O,O'-di-(p-toluoyl)-tartaric acid in the presence of Ca(2+) . Meanwhile, the undesired could be racemized in the presence of sodium hydroxide and the product was suitable for further resolution. A carbanion mechanism for the racemization of is proposed.

  10. Validation of a fully automated robotic setup for preparation of whole blood samples for LC-MS toxicology analysis.

    PubMed

    Andersen, David; Rasmussen, Brian; Linnet, Kristian

    2012-05-01

    A fully automated setup was developed for preparing whole blood samples using a Tecan Evo workstation. By integrating several add-ons to the robotic platform, the flexible setup was able to prepare samples from sample tubes to a 96-well sample plate ready for injection on liquid chromatography-mass spectrometry using several preparation techniques, including protein precipitation, solid-phase extraction and centrifugation, without any manual intervention. Pipetting of a known aliquot of whole blood was achieved by integrating a balance and performing gravimetric measurements. The system was able to handle 1,073 of 1,092 (98.3%) samples of whole blood from forensic material, including postmortem samples, without any need for repeating sample preparation. Only three samples required special treatment such as dilution. The addition of internal and calibration standards were validated by pipetting a solution of Orange G and measuring the weight and absorbance. Internal standard (20 µL) was added in a multi-pipetting sequence with an accuracy of 99.9% and imprecision (coefficient of variation) of 1.6%. Calibration standards were added with high accuracy at volumes as low as 6.00 µL (±0.21 µL). The general setup of the offline sample preparation and key validation parameters of a quantitative analysis of Δ(9)-tetrahydrocannabinol is presented.

  11. Methods for point-of-care detection of nucleic acid in a sample

    DOEpatents

    Bearinger, Jane P.; Dugan, Lawrence C.

    2015-12-29

    Provided herein are methods and apparatus for detecting a target nucleic acid in a sample and related methods and apparatus for diagnosing a condition in an individual. The condition is associated with presence of nucleic acid produced by certain pathogens in the individual.

  12. Apparatus for point-of-care detection of nucleic acid in a sample

    DOEpatents

    Bearinger, Jane P.; Dugan, Lawrence C.

    2016-04-19

    Provided herein are methods and apparatus for detecting a target nucleic acid in a sample and related methods and apparatus for diagnosing a condition in an individual. The condition is associated with presence of nucleic acid produced by certain pathogens in the individual.

  13. Comparison of different sample preparation methods for platinum determination in cultured cells by graphite furnace atomic absorption spectrometry

    PubMed Central

    Cai, Jing; Jia, Jinghui; Zhang, Yuzeng; Dong, Weihong

    2017-01-01

    Background Platinum-based agents are widely used in chemotherapy against solid tumors and insufficient intracellular drug accumulation is one of the leading causes of platinum resistance which is associated with poor survival of tumor patients. Thus, the detection of intracellular platinum is pivotal for studies aiming to overcome platinum resistance. In the present study, we aimed to establish a reliable graphite furnace atomic absorption spectrometry (GFAAS)-based assay to quantify the intracellular platinum content for cultured cells. Methods Several most commonly applied cell preparation methods, including 0.2% HNO3, 0.2% Triton X-100, concentrated nitric acid, RIPA combined with concentrated nitric acid and hydroxide, followed by GFAAS for platinum detection were compared in ovarian, cervical and liver cancer cell lines to obtain the optimal one, and parameters regarding linearity, accuracy, precision and sensitivity were evaluated. Influence of other metals on platinum detection and the storage conditions of samples were also determined. Results The treatment of cells with 0.2% HNO3 was superior to other approaches with fewer platinum loss and better repeatability. The recovery rate and precision of this method were 97.3%–103.0% and 1.4%–3.8%, respectively. The average recoveries in the presence of other metals were 95.1%–103.1%. The detection limit was 13.23 ug/L. The recovery rate of platinum remained acceptable even in cell samples stored in −20 °C or −80 °C for two months. Discussion After comparison, we found that 0.2% HNO3 was optimal for intracellular platinum quantification based on GFAAS, which presented values compatible with that of inductively-coupled plasma mass-spectrometry (ICP-MS), and this is partially attributed to the simplicity of this method. Moreover, the assay was proved to be accurate, sensitive, cost-effective and suitable for the research of platinum-based antitumor therapy. PMID:28123908

  14. Acid and redox properties of mixed oxides prepared by calcination of chromate-containing layered double hydroxides

    SciTech Connect

    Arco, M. del; Carriazo, D.; Martin, C.; Perez-Grueso, A.M.; Rives, V. . E-mail: vrives@usal.es

    2005-11-15

    Layered double hydroxides (LDHs) with Mg and Al in the layers and carbonate, nitrate or chloride in the interlayer, or with Zn and Al in the layers and chloride in the interlayer, have been prepared by coprecipitation, and have been used as precursors to prepare chromate-containing LDHs. All these systems, as well as those obtained upon their calcination up to 800 deg. C, have been characterised by powder X-ray diffraction, FT-IR and vis-UV spectroscopies, temperature-programmed reduction (TPR), nitrogen adsorption at -196 deg. C for surface texture and porosity assessment, and FT-IR monitoring of pyridine adsorption for surface acidity determination. The results obtained show that the crystallinity of the chromate-containing LDH depends on the precursor used. The layered structure of the Mg, Al systems is stabilised up to 400 deg. C upon incorporation of chromate; however, the Zn,Al-chromate samples collapse between 200 and 300 deg. C, with simultaneous formation of ZnO. Calcination of the samples above 400 deg. C gives rise to a reduction of Cr(VI) to Cr(III), as concluded from vis-UV spectroscopic studies. The TPR profiles show that chromate in ZnAl hydrotalcite is more easily reduced than that incorporated in the magnesium ones. Moderately strong surface Lewis acid sites exist in all samples calcined below 500 deg. C.

  15. Acid and redox properties of mixed oxides prepared by calcination of chromate-containing layered double hydroxides

    NASA Astrophysics Data System (ADS)

    del Arco, M.; Carriazo, D.; Martín, C.; Pérez-Grueso, A. M.; Rives, V.

    2005-11-01

    Layered double hydroxides (LDHs) with Mg and Al in the layers and carbonate, nitrate or chloride in the interlayer, or with Zn and Al in the layers and chloride in the interlayer, have been prepared by coprecipitation, and have been used as precursors to prepare chromate-containing LDHs. All these systems, as well as those obtained upon their calcination up to 800 °C, have been characterised by powder X-ray diffraction, FT-IR and vis-UV spectroscopies, temperature-programmed reduction (TPR), nitrogen adsorption at -196 °C for surface texture and porosity assessment, and FT-IR monitoring of pyridine adsorption for surface acidity determination. The results obtained show that the crystallinity of the chromate-containing LDH depends on the precursor used. The layered structure of the Mg, Al systems is stabilised up to 400 °C upon incorporation of chromate; however, the Zn,Al-chromate samples collapse between 200 and 300 °C, with simultaneous formation of ZnO. Calcination of the samples above 400 °C gives rise to a reduction of Cr(VI) to Cr(III), as concluded from vis-UV spectroscopic studies. The TPR profiles show that chromate in ZnAl hydrotalcite is more easily reduced than that incorporated in the magnesium ones. Moderately strong surface Lewis acid sites exist in all samples calcined below 500 °C.

  16. CTEPP STANDARD OPERATING PROCEDURE FOR COLLECTION OF FOOD PREPARATION SURFACE WIPE SAMPLES FOR PERSISTENT ORGANIC POLLUTANTS (SOP-2.17)

    EPA Science Inventory

    This SOP describes the method for collection of the food preparation surface wipe samples for the measurement of persistent organic pollutants (POP). This method uses a wipe to collect POP residues from a surface where a study participant prepares food the most often (i.e., kitch...

  17. Application and comparison of high-speed countercurrent chromatography and high performance liquid chromatography in preparative enantioseparation of α-substitution mandelic acids.

    PubMed

    Tong, Shengqiang; Zhang, Hu; Shen, Mangmang; Ito, Yoichiro; Yan, Jizhong

    2015-04-01

    Preparative enantioseparations of α-cyclopentylmandelic acid and α-methylmandelic acid by high-speed countercurrent chromatography (HSCCC) and high performance liquid chromatography (HPLC) were compared using hydroxypropy-β-cyclodextrin (HP-β-CD) and sulfobutyl ether-β-cyclodextrin (SBE-β-CD) as the chiral mobile phase additives. In preparative HPLC the enantioseparation was achieved on the ODS C18 reverse phase column with the mobile phase composed of a mixture of acetonitrile and 0.10 mol L(-1) phosphate buffer at pH 2.68 containing 20 mmol L(-1) HP-β-CD for α-cyclopentylmandelic acid and 20 mmol L(-1) SBE-β-CD for α-methylmandelic acid. The maximum sample size for α-cyclopentylmandelic acid and α-methylmandelic acid was only about 10 mg and 5 mg, respectively. In preparative HSCCC the enantioseparations of these two racemates were performed with the two-phase solvent system composed of n-hexane-methyl tert.-butyl ether-0.1 molL(-1) phosphate buffer solution at pH 2.67 containing 0.1 mol L(-1) HP-β-CD for α-cyclopentylmandelic acid (8.5:1.5:10, v/v/v) and 0.1 mol L(-1) SBE-β-CD for α-methylmandelic acid (3:7:10, v/v/v). Under the optimum separation conditions, total 250 mg of racemic α-cyclopentylmandelic acid could be completely enantioseparated by HSCCC with HP-β-CD as a chiral mobile phase additive in a single run, yielding 105-110 mg of enantiomers with 95-98% purity and 85-90% recovery. But, no complete enantioseparation of α-methylmandelic acid was achieved by preparative HSCCC with either of the chiral selectors due to their limited enantioselectivity. In this paper preparative enantioseparation by HSCCC and HPLC was compared from various aspects.

  18. Application and comparison of high-speed countercurrent chromatography and high performance liquid chromatography in preparative enantioseparation of α-substitution mandelic acids

    PubMed Central

    Tong, Shengqiang; Zhang, Hu; Shen, Mangmang; Ito, Yoichiro; Yan, Jizhong

    2014-01-01

    Preparative enantioseparations of α-cyclopentylmandelic acid and α-methylmandelic acid by high-speed countercurrent chromatography (HSCCC) and high performance liquid chromatography (HPLC) were compared using hydroxypropy-β-cyclodextrin (HP-β-CD) and sulfobutyl ether-β-cyclodextrin (SBE-β-CD) as the chiral mobile phase additives. In preparative HPLC the enantioseparation was achieved on the ODS C18 reverse phase column with the mobile phase composed of a mixture of acetonitrile and 0.10 mol L−1 phosphate buffer at pH 2.68 containing 20 mmol L−1 HP-β-CD for α-cyclopentylmandelic acid and 20 mmol L−1 SBE-β-CD for α-methylmandelic acid. The maximum sample size for α-cyclopentylmandelic acid and α-methylmandelic acid was only about 10 mg and 5 mg, respectively. In preparative HSCCC the enantioseparations of these two racemates were performed with the two-phase solvent system composed of n-hexane-methyl tert.-butyl ether-0.1 molL−1 phosphate buffer solution at pH 2.67 containing 0.1 mol L−1 HP-β-CD for α-cyclopentylmandelic acid (8.5:1.5:10, v/v/v) and 0.1 mol L−1 SBE-β-CD for α-methylmandelic acid (3:7:10, v/v/v). Under the optimum separation conditions, total 250 mg of racemic α-cyclopentylmandelic acid could be completely enantioseparated by HSCCC with HP-β-CD as a chiral mobile phase additive in a single run, yielding 105-110 mg of enantiomers with 95-98% purity and 85-90% recovery. But, no complete enantioseparation of α-methylmandelic acid was achieved by preparative HSCCC with either of the chiral selectors due to their limited enantioselectivity. In this paper preparative enantioseparation by HSCCC and HPLC was compared from various aspects. PMID:25983356

  19. Modular approach to customise sample preparation procedures for viral metagenomics: a reproducible protocol for virome analysis

    PubMed Central

    Conceição-Neto, Nádia; Zeller, Mark; Lefrère, Hanne; De Bruyn, Pieter; Beller, Leen; Deboutte, Ward; Yinda, Claude Kwe; Lavigne, Rob; Maes, Piet; Ranst, Marc Van; Heylen, Elisabeth; Matthijnssens, Jelle

    2015-01-01

    A major limitation for better understanding the role of the human gut virome in health and disease is the lack of validated methods that allow high throughput virome analysis. To overcome this, we evaluated the quantitative effect of homogenisation, centrifugation, filtration, chloroform treatment and random amplification on a mock-virome (containing nine highly diverse viruses) and a bacterial mock-community (containing four faecal bacterial species) using quantitative PCR and next-generation sequencing. This resulted in an optimised protocol that was able to recover all viruses present in the mock-virome and strongly alters the ratio of viral versus bacterial and 16S rRNA genetic material in favour of viruses (from 43.2% to 96.7% viral reads and from 47.6% to 0.19% bacterial reads). Furthermore, our study indicated that most of the currently used virome protocols, using small filter pores and/or stringent centrifugation conditions may have largely overlooked large viruses present in viromes. We propose NetoVIR (Novel enrichment technique of VIRomes), which allows for a fast, reproducible and high throughput sample preparation for viral metagenomics studies, introducing minimal bias. This procedure is optimised mainly for faecal samples, but with appropriate concentration steps can also be used for other sample types with lower initial viral loads. PMID:26559140

  20. Development of automated preparation system for isotopocule analysis of N2O in various air samples

    NASA Astrophysics Data System (ADS)

    Toyoda, Sakae; Yoshida, Naohiro

    2016-05-01

    Nitrous oxide (N2O), an increasingly abundant greenhouse gas in the atmosphere, is the most important stratospheric ozone-depleting gas of this century. Natural abundance ratios of isotopocules of N2O, NNO molecules substituted with stable isotopes of nitrogen and oxygen, are a promising index of various sources or production pathways of N2O and of its sink or decomposition pathways. Several automated methods have been reported to improve the analytical precision for the isotopocule ratio of atmospheric N2O and to reduce the labor necessary for complicated sample preparation procedures related to mass spectrometric analysis. However, no method accommodates flask samples with limited volume or pressure. Here we present an automated preconcentration system which offers flexibility with respect to the available gas volume, pressure, and N2O concentration. The shortest processing time for a single analysis of typical atmospheric sample is 40 min. Precision values of isotopocule ratio analysis are < 0.1 ‰ for δ15Nbulk (average abundances of 14N15N16O and 15N14N16O relative to 14N14N16O), < 0.2 ‰ for δ18O (relative abundance of 14N14N18O), and < 0.5 ‰ for site preference (SP; difference between relative abundance of 14N15N16O and 15N14N16O). This precision is comparable to that of other automated systems, but better than that of our previously reported manual measurement system.

  1. Using Exclusion-Based Sample Preparation (ESP) to Reduce Viral Load Assay Cost.

    PubMed

    Berry, Scott M; Pezzi, Hannah M; Williams, Eram D; Loeb, Jennifer M; Guckenberger, David J; Lavanway, Alex J; Puchalski, Alice A; Kityo, Cissy M; Mugyenyi, Peter N; Graziano, Franklin M; Beebe, David J

    2015-01-01

    Viral load (VL) measurements are critical to the proper management of HIV in developing countries. However, access to VL assays is limited by the high cost and complexity of existing assays. While there is a need for low cost VL assays, performance must not be compromised. Thus, new assays must be validated on metrics of limit of detection (LOD), accuracy, and dynamic range. Patient plasma samples from the Joint Clinical Research Centre in Uganda were de-identified and measured using both an existing VL assay (Abbott RealTime HIV-1) and our assay, which combines low cost reagents with a simplified method of RNA isolation termed Exclusion-Based Sample Preparation (ESP).71 patient samples with VLs ranging from <40 to >3,000,000 copies/mL were used to compare the two methods. We demonstrated equivalent LOD (~50 copies/mL) and high accuracy (average difference between methods of 0.08 log, R2 = 0.97). Using expenditures from this trial, we estimate that the cost of the reagents and consumables for this assay to be approximately $5 USD. As cost is a significant barrier to implementation of VL testing, we anticipate that our assay will enhance access to this critical monitoring test in developing countries.

  2. Urine sample preparation in 96-well filter plates for quantitative clinical proteomics.

    PubMed

    Yu, Yanbao; Suh, Moo-Jin; Sikorski, Patricia; Kwon, Keehwan; Nelson, Karen E; Pieper, Rembert

    2014-06-03

    Urine is an important, noninvasively collected body fluid source for the diagnosis and prognosis of human diseases. Liquid chromatography mass spectrometry (LC-MS) based shotgun proteomics has evolved as a sensitive and informative technique to discover candidate disease biomarkers from urine specimens. Filter-aided sample preparation (FASP) generates peptide samples from protein mixtures of cell lysate or body fluid origin. Here, we describe a FASP method adapted to 96-well filter plates, named 96FASP. Soluble urine concentrates containing ~10 μg of total protein were processed by 96FASP and LC-MS resulting in 700-900 protein identifications at a 1% false discovery rate (FDR). The experimental repeatability, as assessed by label-free quantification and Pearson correlation analysis for shared proteins among replicates, was high (R ≥ 0.97). Application to urinary pellet lysates which is of particular interest in the context of urinary tract infection analysis was also demonstrated. On average, 1700 proteins (±398) were identified in five experiments. In a pilot study using 96FASP for analysis of eight soluble urine samples, we demonstrated that protein profiles of technical replicates invariably clustered; the protein profiles for distinct urine donors were very different from each other. Robust, highly parallel methods to generate peptide mixtures from urine and other body fluids are critical to increase cost-effectiveness in clinical proteomics projects. This 96FASP method has potential to become a gold standard for high-throughput quantitative clinical proteomics.

  3. Microwave Processing for Sample Preparation to Evaluate Mitochondrial Ultrastructural Damage in Hemorrhagic Shock

    NASA Astrophysics Data System (ADS)

    Josephsen, Gary D.; Josephsen, Kelly A.; Beilman, Greg J.; Taylor, Jodie H.; Muiler, Kristine E.

    2005-12-01

    This is a report of the adaptation of microwave processing in the preparation of liver biopsies for transmission electron microscopy (TEM) to examine ultrastructural damage of mitochondria in the setting of metabolic stress. Hemorrhagic shock was induced in pigs via 35% total blood volume bleed and a 90-min period of shock followed by resuscitation. Hepatic biopsies were collected before shock and after resuscitation. Following collection, biopsies were processed for TEM by a rapid method involving microwave irradiation (Giberson, 2001). Samples pre- and postshock of each of two animals were viewed and scored using the mitochondrial ultrastructure scoring system (Crouser et al., 2002), a system used to quantify the severity of ultrastructural damage during shock. Results showed evidence of increased ultrastructural damage in the postshock samples, which scored 4.00 and 3.42, versus their preshock controls, which scored 1.18 and 1.27. The results of this analysis were similar to those obtained in another model of shock (Crouser et al., 2002). However, the amount of time used to process the samples was significantly shortened with methods involving microwave irradiation.

  4. Urine Sample Preparation in 96-Well Filter Plates for Quantitative Clinical Proteomics

    PubMed Central

    2015-01-01

    Urine is an important, noninvasively collected body fluid source for the diagnosis and prognosis of human diseases. Liquid chromatography mass spectrometry (LC-MS) based shotgun proteomics has evolved as a sensitive and informative technique to discover candidate disease biomarkers from urine specimens. Filter-aided sample preparation (FASP) generates peptide samples from protein mixtures of cell lysate or body fluid origin. Here, we describe a FASP method adapted to 96-well filter plates, named 96FASP. Soluble urine concentrates containing ∼10 μg of total protein were processed by 96FASP and LC-MS resulting in 700–900 protein identifications at a 1% false discovery rate (FDR). The experimental repeatability, as assessed by label-free quantification and Pearson correlation analysis for shared proteins among replicates, was high (R ≥ 0.97). Application to urinary pellet lysates which is of particular interest in the context of urinary tract infection analysis was also demonstrated. On average, 1700 proteins (±398) were identified in five experiments. In a pilot study using 96FASP for analysis of eight soluble urine samples, we demonstrated that protein profiles of technical replicates invariably clustered; the protein profiles for distinct urine donors were very different from each other. Robust, highly parallel methods to generate peptide mixtures from urine and other body fluids are critical to increase cost-effectiveness in clinical proteomics projects. This 96FASP method has potential to become a gold standard for high-throughput quantitative clinical proteomics. PMID:24797144

  5. Preparation and characterization of (10)B boric acid with high purity for nuclear industry.

    PubMed

    Zhang, Weijiang; Liu, Tianyu; Xu, Jiao

    2016-01-01

    Boric acid is often added into coolant as neutron capture agent for pressurized water reactor, whose amount is influenced by its abundance and purity. Therefore, the preparation of enriched (10)B boric acid with high purity is beneficial to nuclear industry. (10)B is also used in developing tumor-specific boronated drugs in boron neutron capture therapy. The boronated drug can be administered to patient intravenously, intratumorally, or deposited at tumor site in surgical excision. Thus, enriched (10)B boric acid is of practical significance in the field of medicine. Self-made boron trifluoride-methanol-complex solution was selected as one of the experimental reagents, and the preparation of (10)B acid was realized by one-step reaction for the complexes with water and calcium chloride. The determination of electrical conductivity in reaction process proves that the optimum reaction time was 16-20 h. Furthermore, the effect of reaction time, ratio of calcium chloride to complex as well as the amount of water on the purity and yield of boric acid was investigated. Finally, the optimum reaction time was 20 h, the optimal solid-liquid ratio (molar ratio) was 3:1, and the amount of water was 1 L of deionized water for each mol of the complex. H2O2 was added in the reaction process to remove Fe(2+). After recrystallization, IR spectra of (10)B boric acid was measured and compared with standard to verify the product of boric acid. The feasibility of the preparation method was determined by the detection of XRD of boric acid. To observe the morphology by polarizing microscope, crystal structure was obtained. The purity of the final product is 99.95 %, and the yield is 96.47 %. The ion concentration of boric acid accords with the national standard of high purity, which was determined by ICP.

  6. Lights Will Guide You : Sample Preparation and Applications for Integrated Laser and Electron Microscopy

    NASA Astrophysics Data System (ADS)

    Karreman, M. A.

    2013-03-01

    Correlative microscopy is the combined use of two different forms of microscopy in the study of a specimen, allowing for the exploitation of the advantages of both imaging tools. The integrated Laser and Electron Microscope (iLEM), developed at Utrecht University, combines a fluorescence microscope (FM) and a transmission electron microscope (TEM) in a single set-up. The region of interest in the specimen is labeled or tagged with a fluorescent probe and can easily be identified within a large field of view with the FM. Next, this same area is retraced in the TEM and can be studied at high resolution. The iLEM demands samples that can be imaged with both FM and TEM. Biological specimen, typically composed of light elements, generate low image contrast in the TEM. Therefore, these samples are often ‘contrasted’ with heavy metal stains. FM, on the other hand, images fluorescent samples. Sample preparation for correlative microscopy, and iLEM in particular, is complicated by the fact that the heavy metals stains employed for TEM quench the fluorescent signal of the probe that is imaged with FM. The first part of this thesis outlines preparation procedures for biological material yielding specimen that can be imaged with the iLEM. Here, approaches for the contrasting of thin sections of cells and tissue are introduced that do not affect the fluorescence signal of the probe that marks the region of interest. Furthermore, two novel procedures, VIS2FIXH and VIS2FIX­FS are described that allow for the chemical fixation of thin sections of cryo-immobilized material. These procedures greatly expedite the sample preparation process, and open up novel possibilities for the immuno-labeling of difficult antigens, eg. proteins and lipids that are challenging to preserve. The second part of this thesis describes applications of iLEM in research in the field of life and material science. The iLEM was employed in the study of UVC induced apoptosis (programmed cell death) of

  7. Effects of sample preparation on the optical properties of breast tissue

    NASA Astrophysics Data System (ADS)

    Marks, Fay A.

    1996-04-01

    The optical properties of biological tissue should be determined in vivo whenever possible. However, for those instances when in vivo studies are impractical, too expensive or inappropriate, and when blood flow is not an issue, the ability to perform in vitro studies then becomes invaluable. Optical absorption spectroscopy shows that it may be possible to obtain meaningful information about the optical properties of human breast tissue from in vitro samples if strict preparation and measuring protocols are used. That a strict protocol for storing and handling tissue is critical can be seen from our observations of changes in the optical absorption spectra that occur in response to formalin fixation, the passage of time, application of stains and dyes, and storage in growth medium of the excised tissue. In vivo optical absorption spectroscopy measurements have been made on human breast cancer xenografts and compared with in vitro measurements on breast biopsies prepared according to precise collection and treatment protocols. There is a 'window of opportunity' before time dependent changes in the UV optical absorption spectra of the excised tissue specimens occur. This time window of opportunity widens at longer wavelengths with the least changes occurring in the optical spectra in the NIR.

  8. Ultrastructure of Plant Leaf Cuticles in relation to Sample Preparation as Observed by Transmission Electron Microscopy

    PubMed Central

    Guzmán, Paula; Fernández, Victoria; García, María Luisa; Fernández, Agustín; Gil, Luis

    2014-01-01

    The leaf cuticular ultrastructure of some plant species has been examined by transmission electron microscopy (TEM) in only few studies. Attending to the different cuticle layers and inner structure, plant cuticles have been grouped into six general morphological types. With the aim of critically examining the effect of cuticle isolation and preparation for TEM analysis on cuticular ultrastructure, adaxial leaf cuticles of blue-gum eucalypt, grey poplar, and European pear were assessed, following a membrane science approach. The embedding and staining protocols affected the ultrastructure of the cuticles analysed. The solubility parameter, surface tension, and contact angles with water of pure Spurr's and LR-White resins were within a similar range. Differences were however estimated for resin : solvent mixtures, since Spurr's resin is combined with acetone and LR-White resin is mixed with ethanol. Given the composite hydrophilic and lipophilic nature of plant cuticles, the particular TEM tissue embedding and staining procedures employed may affect sample ultrastructure and the interpretation of the results in physicochemical and biological terms. It is concluded that tissue preparation procedures may be optimised to facilitate the observation of the micro- and nanostructure of cuticular layers and components with different degrees of polarity and hydrophobicity. PMID:24895682

  9. Recent developments in sample preparation techniques for chromatography analysis of traditional Chinese medicines.

    PubMed

    Deng, Chunhui; Liu, Ning; Gao, Mingxia; Zhang, Xiangmin

    2007-06-15

    Traditional Chinese medicines (TCMs) have a long history dating back thousands of years. Recently, there has been increasing interest worldwide in the use of TCMs for the prevention and treatment of various illnesses. In China, a large number of analytical tools, especially chromatographic techniques have been used to analyze the constituents of TCMs in order to control their quality and discover new bioactive compounds. In this paper, recent developments in sample preparation techniques for the extraction, clean-up, and concentration of analytes from TCMs are compared. These techniques include headspace solid-phase microextraction (HS-SPME), headspace liquid-phase microextraction (HS-LPME), microwave-assisted extraction (MAE), supercritical-fluid extraction (SFE), pressurized-liquid extraction (PLE), and microwave distillation (MD).

  10. Polymer-coated fibrous extraction medium for sample preparation coupled to microcolumn liquid-phase separations.

    PubMed

    Imaizumi, Motohiro; Saito, Yoshihiro; Hayashida, Makiko; Takeichi, Tsutomu; Wada, Hiroo; Jinno, Kiyokatsu

    2003-01-15

    Polymer-coated fibrous material has been introduced as the extraction medium for a miniaturized sample preparation method being coupled with microcolumn liquid chromatography. The preconcentration and the subsequent liquid chromatographic separation of tricyclic antidepressants (TCAs) drugs, amitriptyline, imipramine, nortriptyline and desipramine, was carried out with the hyphenated system. Several basic experimental parameters, such as extraction and separation conditions, were investigated along with the applicability of the method for the analysis of biological fluids. The results clearly showed that the on-line coupled system could be a powerful tool for the analysis of complex mixtures in biological matrix without a large solvent consumption and specially designed instruments. The lowest limit of quantification was quite acceptable for the analysis of TCAs in clinical and forensic situations.

  11. NGSI FY15 Final Report. Innovative Sample Preparation for in-Field Uranium Isotopic Determinations

    SciTech Connect

    Yoshida, Thomas M.; Meyers, Lisa

    2015-11-10

    Our FY14 Final Report included an introduction to the project, background, literature search of uranium dissolution methods, assessment of commercial off the shelf (COTS) automated sample preparation systems, as well as data and results for dissolution of bulk quantities of uranium oxides, and dissolution of uranium oxides from swipe filter materials using ammonium bifluoride (ABF). Also, discussed were reaction studies of solid ABF with uranium oxide that provided a basis for determining the ABF/uranium oxide dissolution mechanism. This report details the final experiments for optimizing dissolution of U3O8 and UO2 using ABF and steps leading to development of a Standard Operating Procedure (SOP) for dissolution of uranium oxides on swipe filters.

  12. Sample processing and cDNA preparation for microbial metatranscriptomics in complex soil communities.

    PubMed

    Carvalhais, Lilia C; Schenk, Peer M

    2013-01-01

    Soil presents one of the most complex environments for microbial communities as it provides many microhabitats that allow coexistence of thousands of species with important ecosystem functions. These include biomass and nutrient cycling, mineralization, and detoxification. Culture-independent DNA-based methods, such as metagenomics, have revealed operational taxonomic units that suggest a high diversity of microbial species and associated functions in soil. An emerging but technically challenging area to profile the functions of microorganisms and their activities is mRNA-based metatranscriptomics. Here, we describe issues and important considerations of soil sample processing and cDNA preparation for metatranscriptomics from bacteria and archaea and provide a set of methods that can be used in the required experimental steps.

  13. Diffuse-reflectance infrared Fourier transform spectroscopy: new technique of sample preparation

    NASA Astrophysics Data System (ADS)

    Hrebičík, M.; Budínová, G.; Godarská, T.; Vláčil, D.; Vogenseh, Stine B.; Volka, K.

    1997-06-01

    A new technique of measurement of the diffuse-reflectance infrared FT spectra, based on the preparation of a cylinder from the mixture of the sample and powdered KBr under pressure of about 5.85 MPa, has been tested. During the measurement, the axis of the formed cylinder is perpendicular to the direction of the incident light. A repeatability of the measurement of selected bands and also of the background was investigated for hydroquinone, nicotinamide, silica gel, rice, tea and also lyophilized human aqueous humour. The relative standard deviations of log( {1}/{R}) showed a dependence on the character of the measured compound, but in general were comparable or slightly better than those obtained by the standard method of loosely packed cups. The values were better than 1.5% in the most cases. The main advantage of the proposed technique lies in its simplicity and rapidity of obtaining statistically significant data.

  14. Phosphite determination in fertilizers after online sequential sample preparation in a flow injection system.

    PubMed

    Dametto, Patrícia Roberta; Franzini, Vanessa Pezza; Gomes Neto, José Anchieta

    2007-07-25

    A flow injection spectrophotometric system is proposed for phosphite determination in fertilizers by the molybdenum blue method after the processing of each sample two times on-line without and with an oxidizing step. The flow system was designed to add sulfuric acid or permanganate solutions alternately into the system by simply displacing the injector-commutator from one resting position to another, allowing the determination of phosphate and total phosphate, respectively. The concentration of phosphite is obtained then by difference between the two measurents. The influence of flow rates, sample volume, and dimension of flow line connecting the injector-commutator to the main analytical channel was evaluated. The proposed method was applied to phosphite determination in commercial liquid fertilizers. Results obtained with the proposed FIA system were not statistically different from those obtained by titrimetry at the 95% confidence level. In addition, recoveries within 94 and 100% of spiked fertilizers were found. The relative standard deviation (n = 12) related to the phosphite-converted-phosphate peak alone was sampling rate was calculated as 15 determinations per hour, and the reagent consumption was about 6.3 mg of KMnO4, 200 mg of (NH4)6Mo7O24.4H2O, and 40 mg of ascorbic acid per measurement.

  15. Evaluation of the effect of surface preparation using phosphoric acid and luting cement on the flexural strength of porcelain laminate veneering material

    PubMed Central

    Guruprasada; Rivankar, N.; Dhiman, R.K.; Viswambaran, M.

    2015-01-01

    Background Conventionally HF acid has been used for etching ceramic veneer restorations before their cementation. Studies are lacking regarding the effectiveness of phosphoric acid as a substitute for HF acid for etching the ceramic veneers. The purpose of this study was to evaluate the effectiveness of surface preparation of porcelain laminate veneers using phosphoric acid, as compared to HF acid etching in providing the necessary surface roughness conducive to development of an effective bond between the ceramic laminate and the resin luting cement. Methods 210 porcelain discs of 15 mm diameter and 0.9 mm thickness were prepared. These study samples were divided into seven groups of thirty samples each. Surfaces of the first (control) and the second group of samples were not prepared. The surfaces of other five groups were prepared with different surface treatments. Further all the groups of specimens were coated with a layer of resin luting cement. Flexural strength of each specimen was determined using universal testing machine and the results were compared. Results The combination surface treatment using alumina surface abrasion followed by etching with phosphoric acid provided the highest flexural strength with the mean flexural strength of 101.11 MPa, followed by alumina surface abrasion (95.41 MPa), and phosphoric acid surface etching (81.68 MPa). Conclusion Laminate veneers surface treated using 50 μm alumina abrasion followed by etching with phosphoric acid showed the highest flexural strengths after resin coating compared to other groups. PMID:26843743

  16. In-line TEM sample preparation and wafer return strategy for rapid yield learning

    NASA Astrophysics Data System (ADS)

    Bicaïs-Lépinay, N.; André, F.; Brevers, S.; Guyader, P.; Trouiller, C.; Kwakman, L. F. Tz.; Pokrant, S.; Verkleij, D.; Schampers, R.; Ithier, L.; Sicurani, E.; Wyon, C.

    2006-03-01

    Full wafer dual beam FIB-SEM systems have received a lot of industrial interest in the last years and by now are operational in several 200mm and 300mm fabs. These tools offer a 3D-physical characterization capability of defects and device structures and as such allow for more rapid yield learning and increased process control. Moreover, if SEM resolution is insufficient to reveal defect origin or the necessary process details, it is now also possible to prepare TEM samples using a controlled, easy to learn in-situ process and to efficiently continue the characterization with a high resolution TEM inspection. Thanks to latest hardware developments and the high degree of automation of this TEM sample preparation process, wafers no longer need to be broken and remain essentially free from contamination. Hence, the TEM lamella process can be considered as non-destructive and wafers may continue the fabrication process flow. In this paper we examine the SEM and TEM application capabilities offered by in-line dual beam systems. To qualify the wafer return strategy, the particle contamination generated by the system hardware as well as the process-induced contamination have been investigated. The particle levels measured are fully acceptable to adopt the wafer return strategy. Ga-contamination does exist but is sufficiently low and localized so that the wafer return strategy can be applied safely in the back-end of line process. Yield analysis has confirmed that there is no measurable impact on device yield. Although yet to be proven for the frond-end of line processes, the wafer return strategy has been demonstrated as a valuable one already in the backend of line processes. The as developed non-destructive 3-D SEM-TEM characterization capability does offer value added data that allow to determine the root cause of critical process defects in almost real-time and this for both standard (SEM) and more advanced (TEM) technologies.

  17. Standardization of a sample preparation and analytical workflow for proteomics of archival endometrial cancer tissue.

    PubMed

    Alkhas, Addie; Hood, Brian L; Oliver, Kate; Teng, Pang-Ning; Oliver, Julie; Mitchell, David; Hamilton, Chad A; Maxwell, G Larry; Conrads, Thomas P

    2011-11-04

    The goal of the present study was to establish a standard operating procedure for mass spectrometry (MS)-based proteomic analysis of laser microdissected (LMD) formalin-fixed, paraffin-embedded (FFPE) uterine tissue. High resolution bioimage analysis of a large endometrial cancer tissue microarray immunostained for the breast cancer type 1 susceptibility protein enabled precise counting of cells to establish that there is an average of 600 cells/nL of endometrial cancer tissue. We sought to characterize the peptide recovery from various volumes of tissue gathered by LMD and processed/digested using the present methodology. We observed a nearly linear increase in peptide recovery amount with increasing tissue volume dissected. There was little discernible difference in the peptide recovery from stromal versus malignant epithelium, and there was no apparent difference in the day-to-day recovery. This methodology reproducibly results in 100 ng of digested peptides per nL of endometrial tissue, or ∼25 pg peptides/endometrial cancer cell. Results from liquid chromatography (LC)-MS/MS experiments to assess the impact of total peptide load on column on the total number of peptides and proteins identified from FFPE tissue digests prepared with the present methodology indicate a demonstrable increase in the total number of peptides identified up to 1000 ng, beyond which diminishing returns were observed. Furthermore, we observed no impact on the peptide identification rates from analyses of equivalent peptide amounts derived from lower volume LMD samples. These results show that this single-tube collection-to-injection proteomics (CTIP) workflow represents a straightforward, scalable, and highly reliable methodology for sample preparation to enable high throughput LMD-MS analysis of tissues derived from biopsy or surgery.

  18. Preparation and Characterization of Nanoliposomes Entrapping Medium-Chain Fatty Acids and Vitamin C by Lyophilization

    PubMed Central

    Yang, Shuibing; Liu, Chengmei; Liu, Wei; Yu, Haixia; Zheng, Huijuan; Zhou, Wei; Hu, Yaqin

    2013-01-01

    The complex nanoliposomes encapsulating both a hydrophilic drug vitamin C (vit C) and hydrophobic drug medium-chain fatty acids (MCFAs) was prepared by combining double emulsion method with dynamic high pressure microfluidization. The complex nanoliposomes was further freeze-dried under −86 °C for 48 h with sucrose at the sucrose/lipids ratio of 2:1(w/w) in order to enhance its stability. The freeze-dried complex nanoliposomes under the suitable conditions exhibited high entrapment efficiency of MCFAs (44.26 ± 3.34)%, relatively high entrapment efficiency of vit C (62.25 ± 3.43)%, low average size diameter (110.4 ± 7.28) nm and good storage stability at 4 °C for 60 days with slight changes in mean particle diameter and drug entrapment efficiencies. The results of transmission electron microscopy of freeze-dried complex nanoliposomes also showed that the freeze-dried samples with sucrose were stable without great increase in their particle sizes and without destroying their spherical shape. The results indicated that sucrose presented well protection effects in MCFAs-vit C complex nanoliposomes, suggesting the possibility of further usage in commercial liposomes. PMID:24084723

  19. Preparation of water and ice samples for 39Ar dating by atom trap trace analysis (ATTA)

    NASA Astrophysics Data System (ADS)

    Schwefel, R.; Reichel, T.; Aeschbach-Hertig, W.; Wagenbach, D.

    2012-04-01

    Atom trap trace analysis (ATTA) is a new and promising method to measure very rare noble gas radioisotopes in the environment. The applicability of this method for the dating of very old groundwater with 81Kr has already been demonstrated [1]. Recent developments now show its feasibility also for the analysis of 39Ar [2,3], which is an ideal dating tracer for the age range between 50 and 1000 years. This range is of interest in the fields of hydro(geo)logy, oceanography, and glaciology. We present preparation (gas extraction and Ar separation) methods for groundwater and ice samples for later analysis by the ATTA technique. For groundwater, the sample size is less of a limitation than for applications in oceanography or glaciology. Large samples are furthermore needed to enable a comparison with the classical method of 39Ar detection by low-level counting. Therefore, a system was built that enables gas extraction from several thousand liters of water using membrane contactors. This system provides degassing efficiencies greater than 80 % and has successfully been tested in the field. Gas samples are further processed to separate a pure Ar fraction by a gas-chromatographic method based on Li-LSX zeolite as selective adsorber material at very low temperatures. The gas separation achieved by this system is controlled by a quadrupole mass spectrometer. It has successfully been tested and used on real samples. The separation efficiency was found to be strongly temperature dependent in the range of -118 to -130 °C. Since ATTA should enable the analysis of 39Ar on samples of less than 1 ccSTP of Ar (corresponding to about 100 ml of air, 2.5 l of water or 1 kg of ice), a method to separate Ar from small amounts of gas was developed. Titanium sponge was found to absorb 60 ccSTP of reactive gases per g of the getter material with reasonably high absorption rates at high operating temperatures (~ 800 ° C). Good separation (higher than 92 % Ar content in residual gas) was

  20. Carbon Isotopic Measurements of Amino Acids in Stardust-Returned Samples

    NASA Technical Reports Server (NTRS)

    Elsila, Jamie

    2009-01-01

    NASA's Stardust spacecraft returned to Earth samples from comet 81P/Wild 2 in January 2006. Preliminary examinations revealed the presence of a suite of organic compounds including several amines and amino acids, but the origin of these compounds could not be identified. Here, we present the carbon isotopic ratios of glycine and e-aminocaproic acid (EACA), the two most abundant amino acids, in Stardust-returned foil samples measured by gas chromatography-combustion-isotope ratio mass spectrometry coupled with quadrupole mass spectrometry (GC-CAMS/IRMS).

  1. Carbon Isotopic Ratios of Amino Acids in Stardust-Returned Samples

    NASA Technical Reports Server (NTRS)

    Elsila, Jamie E.; Glavin, Daniel P.; Dworkin, Jason P.

    2009-01-01

    NASA's Stardust spacecraft returned to Earth samples from comet 81P/Wild 2 in January 2006. Preliminary examinations revealed the presence of a suite of organic compounds including several amines and amino acids, but the origin of these compounds could not be identified. Here. we present the carbon isotopic ratios of glycine and E-aminocaproic acid (EACH), the two most abundant amino acids observed, in Stardust-returned foil samples measured by gas chromatography-combustion-isotope ratio crass spectrometry coupled with quadrupole mass spectrometry (GC-QMS/IRMS).

  2. Fuel properties of heptadecene isomers prepared via tandem isomerization-decarboxylation of oleic acid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Heptadecene isomers were prepared via tandem isomerization-decarboxylation of oleic acid using catalytic triruthenium dodecacarbonyl [Ru3(CO)12]. Chromatographic and spectroscopic characterization of the isolated heptadecene mixture indicated that it consisted of 96% internal trans isomers and 4% ar...

  3. Electrochemical determination of hydrochlorothiazide and folic acid in real samples using a modified graphene oxide sheet paste electrode.

    PubMed

    Beitollahi, Hadi; Hamzavi, Mozhdeh; Torkzadeh-Mahani, Masoud

    2015-01-01

    A new ferrocene-derivative compound, 2-chlorobenzoyl ferrocene, was synthesized and used to construct a modified graphene oxide sheet paste electrode. The electrooxidation of hydrochlorothiazide at the surface of the modified electrode was studied. Under optimized conditions, the square wave voltammetric (SWV) peak current of hydrochlorothiazide increased linearly with hydrochlorothiazide concentration in the range of 5.0 × 10(-8) to 2.0 × 10(-4) M and a detection limit of 20.0 nM was obtained for hydrochlorothiazide. The diffusion coefficient and kinetic parameters (such as electron transfer coefficient and the heterogeneous rate constant) for hydrochlorothiazide oxidation were also determined. The prepared modified electrode exhibits a very good resolution between the voltammetric peaks of hydrochlorothiazide and folic acid which makes it suitable for the detection of hydrochlorothiazide in the presence of folic acid in real samples.

  4. Advanced sample preparation for the molecular quantification of Staphylococcus aureus in artificially and naturally contaminated milk.

    PubMed

    Aprodu, Iuliana; Walcher, Georg; Schelin, Jenny; Hein, Ingeborg; Norling, Börje; Rådström, Peter; Nicolau, Anca; Wagner, Martin

    2011-03-01

    Sample treatment is an essential element when using real-time PCR for quantification of pathogens directly on food samples. This study comparatively evaluated three different principles of sample treatment, i.e. immunomagnetic separation based on phage-derived cell wall binding molecules, matrix solubilization and flotation, in order to establish their suitability for quantifying low numbers of Staphylococcus aureus in milk. All three procedures succeeded to remove S. aureus from the milk matrix, either raw or pasteurized, and, as a result of the concentration of the target cells, minimized the effect of milk associated PCR inhibitors. Sample preparation based on immunomagnetic separation albeit of being user friendly, specific and rapid, failed to allow quantification of low and medium numbers (<10(4)CFU) of S. aureus. In a mastitic milk model cell wall binding domain (CBD)-based target cell extraction revealed results most closely matching those derived from culture-based quantification. Both matrix lysis and flotation allowed quantification of S. aureus at a level of 1-10 cells per ml. Both methods resulted in higher numbers of bacterial cell equivalents (bce) than plating could reveal. Since both methods harvest cells that have been subjected to either mechanical and chemical stresses before quantification, we concluded that the higher bce numbers resulted from a disaggregation of S. aureus clusters initially present in the inoculum. Conclusively, since likely each S. aureus cell of a toxigenic strain contributes to enterotoxin production, molecular quantification could provide an even more realistic impact assessment in outbreak investigations than plating does.

  5. Second generation laser-heated microfurnace for the preparation of microgram-sized graphite samples

    NASA Astrophysics Data System (ADS)

    Yang, Bin; Smith, A. M.; Long, S.

    2015-10-01

    We present construction details and test results for two second-generation laser-heated microfurnaces (LHF-II) used to prepare graphite samples for Accelerator Mass Spectrometry (AMS) at ANSTO. Based on systematic studies aimed at optimising the performance of our prototype laser-heated microfurnace (LHF-I) (Smith et al., 2007 [1]; Smith et al., 2010 [2,3]; Yang et al., 2014 [4]), we have designed the LHF-II to have the following features: (i) it has a small reactor volume of 0.25 mL allowing us to completely graphitise carbon dioxide samples containing as little as 2 μg of C, (ii) it can operate over a large pressure range (0-3 bar) and so has the capacity to graphitise CO2 samples containing up to 100 μg of C; (iii) it is compact, with three valves integrated into the microfurnace body, (iv) it is compatible with our new miniaturised conventional graphitisation furnaces (MCF), also designed for small samples, and shares a common vacuum system. Early tests have shown that the extraneous carbon added during graphitisation in each LHF-II is of the order of 0.05 μg, assuming 100 pMC activity, similar to that of the prototype unit. We use a 'budget' fibre packaged array for the diode laser with custom built focusing optics. The use of a new infrared (IR) thermometer with a short focal length has allowed us to decrease the height of the light-proof safety enclosure. These innovations have produced a cheaper and more compact device. As with the LHF-I, feedback control of the catalyst temperature and logging of the reaction parameters is managed by a LabVIEW interface.

  6. Study of ion beam damage in magnetic tunnel junction on FIB prepared samples

    NASA Astrophysics Data System (ADS)

    Park, Kwangho; Yang, Cheolwoong; Kim, Kyuchul; Nam, Dongwoo; Hwang, Kyuman; Bae, Junsoo; Ahn, Juhyeon; Choi, Jin; Park, Soonoh; Jeong, Sangsup; Cho, Hanku; Jeong, Eunseung

    2013-09-01

    Magnetic Random Access Memory (MRAM) has emerged as the leading candidate for future universal memory due to its non-volatility, excellent endurance and read/write performance. The magnetic tunnel junction (MTJ) is a data storage element in MRAM and is basically composed of two ferromagnetic layers separated by the magnesium oxide (MgO) tunnel barrier. MgO between two ferromagnetic layers was adopted to enlarge the resistance difference between two kinds of magnetic arrangements by tunneling current through MgO. Like this, it is important to understand characterization of MgO for developing Mram. Due to thin thickness of MgO, FIB milling should be used for the preparation of TEM specimens in Mram. The major problem in MgO sampling by FIB milling is the transform of MgO between two ferromagnetic due to FIB induced damage, which leads to high tunnel current through MgO and high resistance difference between two kinds of magnetic arrangements. An understanding of FIB generated artifact on MgO is important to analysis Mram and to optimize the sample preparation process. The normal ion beam damage are compared with low-keV FIB ion beam damage on blanket MgO wafer. Experiments were performed using Helios 450 FIB(FEI) and XV-200TBs(SII) with gallium ion sources operated at 30 keV to 2 keV, respectively. As a preliminary, the thicknesses of all specimens were fixed at 100nm for the final ion beam milling currents of 210 pA(30 keV) by Helios 450 FIB(FEI). Specimens of 100nm were transferred to low-keV FIB (Helios 450/XV-200TBs) to do the low-keV ion milling. Then each specimen had a 2 keV cleaned surface and a 30 keV FIB prepared surface. In this paper, we understand the normal ion beam damage on blanket MgO through changing beam current and beam voltage. Then we present the optimized recipe and which equipment is better to analysis.

  7. High surface area porous carbons prepared from hydrochars by phosphoric acid activation.

    PubMed

    Wang, Lili; Guo, Yupeng; Zou, Bo; Rong, Chunguang; Ma, Xiaoyu; Qu, Yuning; Li, Ying; Wang, Zichen

    2011-01-01

    In the present work, a new route for preparation of high-performance porous carbons under mild conditions was reported. The high surface area (2700 m2/g) and large pore volume (1.98 cm3/g) porous carbons were prepared from hydrochars by conventional phosphoric acid activation method. The hydrochars described here can be obtained from sulfuric acid hydrolysis of rice husk via dehydration, polymerization and carbonization. A specific capacitance of 130 F g(-1) was achieved by using the porous carbon, indicating that the porous carbon prepared by this route has good electrochemical performance. Furthermore, the localized graphitic nature of the porous carbon was proved by X-ray diffraction pattern.

  8. Preparation and evaluation of advanced catalysts for phosphoric acid fuel cells

    NASA Technical Reports Server (NTRS)

    Stonehart, P.; Baris, J.; Hockmuth, J.; Pagliaro, P.

    1984-01-01

    The platinum electrocatalysts were characterized for their crystallite sizes and the degree of dispersion on the carbon supports. One application of these electrocatalysts was for anodic oxidation of hydrogen in hot phosphoric acid fuel cells, coupled with the influence of low concentrations of carbon monoxide in the fuel gas stream. In a similar way, these platinum on carbon electrocatalysts were evaluated for oxygen reduction in hot phosphoric acid. Binary noble metal alloys were prepared for anodic oxidation of hydrogen and noble metal-refractory metal mixtures were prepared for oxygen reduction. An exemplar alloy of platinum and palladium (50/50 atom %) was discovered for anodic oxidation of hydrogen in the presence of carbon monoxide, and patent disclosures were submitted. For the cathode, platinum-vanadium alloys were prepared showing improved performance over pure platinum. Preliminary experiments on electrocatalyst utilization in electrode structures showed low utilization of the noble metal when the electrocatalyst loading exceeded one weight percent on the carbon.

  9. Preparation and in vitro evaluation of a pluronic lecithin organogel containing ricinoleic acid for transdermal delivery.

    PubMed

    Boddu, Sai Hs; Bonam, Sindhu Prabha; Wei, Yangjie; Alexander, Kenneth

    2014-01-01

    The present study deals with the preparation and in vitro evaluation of a Pluronic lecithin organogel gel containing ricinoleic acid for transdermal delivery. Blank Pluronic lecithin organogel gels were prepared using ricinoleic acid as the oil phase and characterized for pH, viscosity, gelation temperature, and microscopic structure. The optimized Pluronic lecithin organogel gel formulation was further evaluated using ketoprofen (10%) and dexamethasone (0.5%) as model drugs. The stability and in vitro permeability of ketoprofen and dexamethasone was evaluated and compared with the corresponding control formulation (Pluronic lecithin organogel gel made with isopropyl palmitate as the oil phase). The pH and viscosity of blank Pluronic lecithin organogel gel prepared with ricinoleic acid was comparable with the isopropyl palmitate Pluronic lecithin organogel gel. The thixotropic property of ricinoleic acid Pluronic lecithin organogel gel was found to be better than the control. Drug-loaded Pluronic lecithin organogel gels behaved in a similar manner and all formulations were found to be stable at 25 degrees C, 35 degrees C, and 40 degrees C for up to 35 days. The penetration profile of dexamethasone was similar from both the Pluronic lecithin organogel gels, while the permeability for ketoprofen from Pluronic lecithin organogel gel containing ricinoleic acid was found to be three times higher as compared to the control formulation.

  10. Monoterpene-based chiral β-amino acid derivatives prepared from natural sources: syntheses and applications.

    PubMed

    Szakonyi, Zsolt; Fülöp, Ferenc

    2011-08-01

    Natural monoterpenes have proved to be good starting materials for the synthesis of β-amino acid derivatives. In the past decade, a number of well-known synthetic procedures have been applied for the preparation of monoterpene-based β-amino acid derivatives, e.g. from β-lactams via the 1,2-dipolar cycloaddition of chlorosulfonyl isocyanate to commercial or readily available monoterpenes [e.g. (+)- and (-)-α- or δ-pinene, (+)-3- and 2-carene, (+)- and (-)-apopinene], the conjugate addition of amides to monoterpene-based α,β-unsaturated esters or the transformations of (-)-cis-pinonoic acid prepared by the oxidative cleavage of (+)- and (-)-verbenone. β-Amino acid derivatives are excellent building blocks for versatile transformations, e.g. multicomponent reactions resulting in β-lactams, syntheses of 1,3-heterocycles and diaminopyrimidine derivatives or the formation of peptides containing an H12 helix. 1,3-Amino alcohol derivatives prepared from β-amino esters have been applied as chiral catalysts in enantioselective transformations. Several of these compounds are of noteworthy pharmacological importance, such as tyrosine kinase Axl inhibitor diaminopyrimidine-coupled β-aminocarboxamides, MDR inhibitor thiourea derivatives of β-amino esters or 2-imino-1,3-oxazines, which exhibit marked growth inhibitory activity on multiple cancer cell lines. The present review summarizes recent developments relating to the syntheses, applications and pharmaceutical importance of monoterpene-based β-amino acids and their derivatives.

  11. Data supporting the rat brain sample preparation and validation assays for simultaneous determination of 8 neurotransmitters and their metabolites using liquid chromatography-tandem mass spectrometry.

    PubMed

    Wojnicz, Aneta; Ortiz, José Avendaño; Casas, Ana I; Freitas, Andiara E; López, Manuela G; Ruiz-Nuño, Ana

    2016-06-01

    The data presented in this article supports the rat brain sample preparation procedure previous to its injection into the liquid chromatography-tandem mass spectrometry (LC-MS/MS) system to monitor levels of adrenaline, noradrenaline, glutamic acid, γ-aminobutyric acid, dopamine, 5-hydroxytryptamine, 5-hydroxyindole acetic acid, and 3-methoxy-4-hydroxyphenylglycol. In addition, we describe the method validation assays (such as calibration curve, lower limit of quantification, precision and accuracy intra- and inter-day, selectivity, extraction recovery and matrix effect, stability, and carry-over effect) according to the United States Food and Drug Administration and European Medicine Agency to measure in one step different neurotransmitters and their metabolites. The data supplied in this article is related to the research study entitled: "Simultaneous determination of 8 neurotransmitters and their metabolite levels in rat brain using liquid chromatography in tandem with mass spectrometry: application to the murine Nrf2 model of depression" (Wojnicz et al. 2016) [1].

  12. Optical biosensor system with integrated microfluidic sample preparation and TIRF based detection

    NASA Astrophysics Data System (ADS)

    Gilli, Eduard; Scheicher, Sylvia R.; Suppan, Michael; Pichler, Heinz; Rumpler, Markus; Satzinger, Valentin; Palfinger, Christian; Reil, Frank; Hajnsek, Martin; Köstler, Stefan

    2013-05-01

    There is a steadily growing demand for miniaturized bioanalytical devices allowing for on-site or point-of-care detection of biomolecules or pathogens in applications like diagnostics, food testing, or environmental monitoring. These, so called labs-on-a-chip or micro-total analysis systems (μ-TAS) should ideally enable convenient sample-in - result-out type operation. Therefore, the entire process from sample preparation, metering, reagent incubation, etc. to detection should be performed on a single disposable device (on-chip). In the early days such devices were mainly fabricated using glass or silicon substrates and adapting established fabrication technologies from the electronics and semiconductor industry. More recently, the development focuses on the use of thermoplastic polymers as they allow for low-cost high volume fabrication of disposables. One of the most promising materials for the development of plastic based lab-on-achip systems are cyclic olefin polymers and copolymers (COP/COC) due to their excellent optical properties (high transparency and low autofluorescence) and ease of processing. We present a bioanalytical system for whole blood samples comprising a disposable plastic chip based on TIRF (total internal reflection fluorescence) optical detection. The chips were fabricated by compression moulding of COP and microfluidic channels were structured by hot embossing. These microfluidic structures integrate several sample pretreatment steps. These are the separation of erythrocytes, metering of sample volume using passive valves, and reagent incubation for competitive bioassays. The surface of the following optical detection zone is functionalized with specific capture probes in an array format. The plastic chips comprise dedicated structures for simple and effective coupling of excitation light from low-cost laser diodes. This enables TIRF excitation of fluorescently labeled probes selectively bound to detection spots at the microchannel surface

  13. Tank 12H Acidic Chemical Cleaning Sample Analysis And Material Balance

    SciTech Connect

    Martino, C. J.; Reboul, S. H.; Wiersma, B. J.; Coleman, C. J.

    2013-11-08

    A process of Bulk Oxalic Acid (BOA) chemical cleaning was performed for Tank 12H during June and July of 2013 to remove all or a portion of the approximately 4400 gallon sludge heel. Three strikes of oxalic acid (nominally 4 wt% or 2 wt%) were used at 55°C and tank volumes of 96- to 140-thousand gallons. This report details the sample analysis of a scrape sample taken prior to BOA cleaning and dip samples taken during BOA cleaning. It also documents a rudimentary material balance for the Tank 12H cleaning results.

  14. Proteomic tools for environmental microbiology--a roadmap from sample preparation to protein identification and quantification.

    PubMed

    Wöhlbrand, Lars; Trautwein, Kathleen; Rabus, Ralf

    2013-10-01

    The steadily increasing amount of (meta-)genomic sequence information of diverse organisms and habitats has a strong impact on research in microbial physiology and ecology. In-depth functional understanding of metabolic processes and overall physiological adaptation to environmental changes, however, requires application of proteomics, as the context specific proteome constitutes the true functional output of a cell. Considering the enormous structural and functional diversity of proteins, only rational combinations of various analytical approaches allow a holistic view on the overall state of the cell. Within the past decade, proteomic methods became increasingly accessible to microbiologists mainly due to the robustness of analytical methods (e.g. 2DE), and affordability of mass spectrometers and their relative ease of use. This review provides an overview on the complex portfolio of state-of-the-art proteomics and highlights the basic principles of key methods, ranging from sample preparation of laboratory or environmental samples, via protein/peptide separation (gel-based or gel-free) and different types of mass spectrometric protein/peptide analyses, to protein identification and abundance determination.

  15. Site-Specific Cryo-focused Ion Beam Sample Preparation Guided by 3D Correlative Microscopy

    PubMed Central

    Arnold, Jan; Mahamid, Julia; Lucic, Vladan; de Marco, Alex; Fernandez, Jose-Jesus; Laugks, Tim; Mayer, Tobias; Hyman, Anthony A.; Baumeister, Wolfgang; Plitzko, Jürgen M.

    2016-01-01

    The development of cryo-focused ion beam (cryo-FIB) for the thinning of frozen-hydrated biological specimens enabled cryo-electron tomography (cryo-ET) analysis in unperturbed cells and tissues. However, the volume represented within a typical FIB lamella constitutes a small fraction of the biological specimen. Retaining low-abundance and dynamic subcellular structures or macromolecular assemblies within such limited volumes requires precise targeting of the FIB milling process. In this study, we present the development of a cryo-stage allowing for spinning-disk confocal light microscopy at cryogenic temperatures and describe the incorporation of the new hardware into existing workflows for cellular sample preparation by cryo-FIB. Introduction of fiducial markers and subsequent computation of three-dimensional coordinate transformations provide correlation between light microscopy and scanning electron microscopy/FIB. The correlative approach is employed to guide the FIB milling process of vitrified cellular samples and to capture specific structures, namely fluorescently labeled lipid droplets, in lamellas that are 300 nm thick. The correlation procedure is then applied to localize the fluorescently labeled structures in the transmission electron microscopy image of the lamella. This approach can be employed to navigate the acquisition of cryo-ET data within FIB-lamellas at specific locations, unambiguously identified by fluorescence microscopy. PMID:26769364

  16. Preparation of higher-actinide burnup and cross section samples. [LMFBR

    SciTech Connect

    Adair, H.L.; Kobisk, E.H.; Quinby, T.C.; Thomas, D.K.; Dailey, J.M.

    1981-01-01

    A joint research program involving the United States and the United Kingdom was instigated about four years ago for the purpose of studying burnup of higher actinides using in-core irradiation in the fast reactor at Dounreay, Scotland. Simultaneously, determination of cross sections of a wide variety of higher actinide isotopes was proposed. Coincidental neutron flux and energy spectral measurements were to be made using vanadium encapsulated dosimetry materials in the immediate region of the burnup and cross section samples. The higher actinide samples chosen for the burnup study were /sup 241/Am and /sup 244/Cm in the forms of Am/sub 2/O/sub 3/, Cm/sub 2/O/sub 3/, and Am/sub 6/ Cm(RE)/sub 7/O/sub 21/, where (RE) represents a mixture of lanthanide sesquioxides. It is the purpose of this paper to describe technology development and its application in the preparation of the fuel specimens and the cross section specimens that are being used in this cooperative program.

  17. Sample preparation for metalloprotein analysis: A case study using horse chestnuts.

    PubMed

    de Magalhães, Cristiana Schmidt; Arruda, Marco Aurélio Zezzi

    2007-03-30

    In the present work, 11 different procedures for protein and metalloprotein extraction from horse chestnuts (Aescullus hippocastanum L.) in natura were tested. After each extraction, total protein was determined and, after protein separation through sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), those metals belonging to the protein structure were mapped by synchrotron radiation X-ray fluorescence (SRXRF). After mapping the elements (Cr, Fe and Mn) in the protein bands (ca. 33 and 23.7kDa), their concentrations were determined using atomic absorption spectrometry (ET AAS). Good results were obtained for protein extraction using a combination of grinding and sonication. However, this strategy was not suitable to preserve metal ions in the protein structure. In fact, there was 42% decrease on Mn concentration using this procedure, compared to that performed with sample agitation in water (taken as reference). On the other hand, when grinding and agitation with an extracting buffer was used, there was a 530% increase of Mn concentration, when compared to the reference procedure. These results indicate agreement between metal identification and determination in proteins as well as the great influence of the extraction procedure (i.e., the sample preparation step) for preserving metals in the protein structures.

  18. A cost-effective plate-based sample preparation for antibody N-glycan analysis.

    PubMed

    Burnina, Irina; Hoyt, Erik; Lynaugh, Heather; Li, Huijuan; Gong, Bing

    2013-09-13

    During early cell line and process development of therapeutic antibodies, a cost-effective high-throughput approach to characterize the N-linked glycans is highly desired given that a large number of samples need to be analyzed. Using commercially available, low cost 96-well plates, we developed a practical procedure to prepare fluorescently labeled N-linked glycans for both qualitative and quantitative analysis by mass spectrometry (MS) and ultrahigh performance liquid chromatography (UPLC). Antibody samples were continuously denatured, reduced, and deglycosylated in a single 96-well hydrophobic membrane filter plate. Subsequently, released glycans were fluorescently labeled in a collection plate, and cleaned-up using a hydrophilic membrane filter plate. Carried out entirely in ready-to-use 96-well plates with simple buffer systems, this procedure requires less than 90min to finish. We applied the optimized procedure to examine the N-linked glycosylation of trastuzumab and were able to quantify ten major N-linked glycans. The results from different amounts of starting materials (10-200μg) were highly similar and showed the robustness of this procedure. Compared to other methods, this new procedure is simple to implement, economically more affordable, and could be very valuable for early screenings of antibody development.

  19. Characteristics of microporous/mesoporous carbons prepared from rice husk under base- and acid-treated conditions.

    PubMed

    Liou, Tzong-Horng; Wu, Shao-Jung

    2009-11-15

    The study reports the preparation of activated carbon with a high surface area from rice husk using chemical activation with H(3)PO(4) and ZnCl(2). Activated carbon prepared from rice husk usually exhibits low specific surface areas due to its high ash content. However, experimental results show that base-leaching and acid-washing processes can effectively enhance the adsorption capacity of rice-husk carbon. The study also investigates the effects of preparation parameters on the surface characteristics of the carbon. These parameters include the kind of activating agent, before and after treatment procedures, impregnation ratio and activation temperature. The chemical and physical properties of samples were examined by EA, ICP-MS, XRD, FTIR, SEM and a N(2)-adsorption meter. The surface areas obtained from ZnCl(2) and H(3)PO(4) activation are as high as 2434 and 1741 m(2)/g, respectively. These values are higher than that of activated carbon treated with neither base nor acid (1262 and 508 m(2)/g for ZnCl(2) and H(3)PO(4) activation). Thermogravimetric analysis shows that the activation process can be divided into three parts based on temperature zones. The results of this study will be useful in developing resource recovery systems for agricultural biomass.

  20. Amino acid analysis in micrograms of meteorite sample by nanoliquid chromatography-high-resolution mass spectrometry.

    PubMed

    Callahan, Michael P; Martin, Mildred G; Burton, Aaron S; Glavin, Daniel P; Dworkin, Jason P

    2014-03-07

    Amino acids and their enantiomers in a 360 microgram sample of Murchison meteorite were unambiguously identified and quantified using chemical derivatization and nanoliquid chromatography coupled to nanoelectrospray ionization high resolution orbitrap mass spectrometry techniques. The distribution and abundance of amino acids were similar to past studies of Murchison meteorite but the samples used here were three orders of magnitude lower. The analytical method was also highly sensitive, and some amino acid reference standards were successfully detected at a level of ∼200 attomoles (on column). These results may open up the possibility for investigating other less studied, sample-limited extraterrestrial samples (e.g., micrometeorites, interplanetary dust particles, and cometary particles) for biologically-relevant organic molecules.

  1. Multiple stable isotope characterization as a forensic tool to distinguish acid scavenger samples

    SciTech Connect

    Moran, James J.; Kreuzer, Helen W.; Carman, April J.; Wahl, Jon H.; Duckworth, Douglas C.

    2012-01-01

    Acid scavengers are frequently used as stabilizer compounds in a variety of applications. When used to stabilize volatile compounds such as nerve agents, the lower volatility and higher stability of acid scavengers make them more persistent in a post-event forensic setting. We are employing compound-specific stable isotope analysis of the carbon, nitrogen, and hydrogen components of three acid scavenging compounds (N,N-diethylaniline, tributylamine, and triethylamine) as a tool for distinguishing between different samples of the stabilizers. Combined analysis of three stable isotopes in these samples improves the technique’s resolving potential, enhancing sample matching capabilities. The compound specific methods developed here can be applied to instances where these compounds are not pure, such as when mixed with an agent or when found as a residue at an event site. Effective sample matching can be crucial for linking compounds at multiple event sites or linking a supply inventory to an event.

  2. Effect of sample preparation on contaminant leaching from copper smelting slag.

    PubMed

    Vítková, Martina; Ettler, Vojtěch; Mihaljevič, Martin; Sebek, Ondřej

    2011-12-15

    Currently standardised leaching tests require grain size reduction, which for large fragments of slags could overestimate the leaching results. To assess the effect of the fine-grained fraction generated by sample crushing, a set of leaching experiments was performed on copper smelting slag from the Zambian Copperbelt: (i) EN 12457-2 batch tests (standardised grain size <4 mm; modified procedure with grain size of 4-0.5 mm simulating exposure of larger fragments on the dumps) and (ii) CEN/TS 14997 pH-static tests (standardised grain size <1 mm simulating the possible wind dispersion scenario near the slag crushing facilities or disposal of fine-grained granulated slag; additional grain sizes <5 mm, 5-0.5 mm and 5-0.5 mm after ultrasonic cleaning). A higher proportion of the fine-grained fraction generally led to greater leaching of Cu, Co and Zn. The metal levels in the leachates under circum-neutral conditions were all below the EU limits for non-hazardous waste. However, at pH 4, the presence of fine dust particles dramatically increased the concentrations of metals in the slag leachates. The greater leachability of Cu and Co from slag particles under acidic conditions suggests a risk of their mobilisation in acidic soils in the Copperbelt area.

  3. Low-temperature preparation of highly conductive thin films from acrylic acid-stabilized silver nanoparticles prepared through ligand exchange.

    PubMed

    Vo, Duc Quy; Shin, Eun Woo; Kim, Jae-Seong; Kim, Sunwook

    2010-11-16

    The preparation of AcA-stabilized Ag nanoparticles and its application to make highly conductive thin films are reported. The AcA-stabilized Ag nanoparticles were prepared through a ligand exchange of original oleylamine (OLA)-coated Ag nanoparticles with acrylic acid (AcA), which acted as both an antisolvent and a modifying ligand during the ligand exchange process. Efficiencies of the ligand exchange as well as the properties of Ag nanoparticles were analyzed using various techniques including TEM, FT-IR, XPS, TGA, and UV-vis methods. The thin films were fabricated by annealing spin-coated AcA-stabilized Ag nanoparticles. Further, the effects of annealing temperature, time, and film thickness on both the film morphology and electrical conductivity have been investigated. In this work, due to the low boiling temperature of stabilizer (AcA) and adjustment of annealing conditions, high electrical conductivity was obtained for the Ag thin films. For example, when annealing at 175 °C for 30 min, a 70 nm thick film showed a maximum electrical conductivity of 1.12 × 10(5) S cm(-1). A conductive layer on a flexible polymer substrate (e.g., PET) sheet has been successfully prepared by annealing a spin-coated film at 140 °C for 30 min. The combined advantages of long-term stability of the AcA-stabilized Ag nanoparticles, low annealing temperature, and high conductivity of the prepared thin films make this relatively simple method attractive for applications in flexible electronics.

  4. Applicability Comparison of Methods for Acid Generation Assessment of Rock Samples

    NASA Astrophysics Data System (ADS)

    Oh, Chamteut; Ji, Sangwoo; Yim, Giljae; Cheong, Youngwook

    2014-05-01

    Minerals including various forms of sulfur could generate AMD (Acid Mine Drainage) or ARD (Acid Rock Drainage), which can have serious effects on the ecosystem and even on human when exposed to air and/or water. To minimize the hazards by acid drainage, it is necessary to assess in advance the acid generation possibility of rocks and estimate the amount of acid generation. Because of its relatively simple and effective experiment procedure, the method of combining the results of ABA (Acid Base Accounting) and NAG (Net Acid Generation) tests have been commonly used in determining acid drainage conditions. The simplicity and effectiveness of the above method however, are derived from massive assumptions of simplified chemical reactions and this often leads to results of classifying the samples as UC (Uncertain) which would then require additional experimental or field data to reclassify them properly. This paper therefore, attempts to find the reasons that cause samples to be classified as UC and suggest new series of experiments where samples can be reclassified appropriately. Study precedents on evaluating potential acid generation and neutralization capacity were reviewed and as a result three individual experiments were selected in the light of applicability and compatibility of minimizing unnecessary influence among other experiments. The proposed experiments include sulfur speciation, ABCC (Acid Buffering Characteristic Curve), and Modified NAG which are all improved versions of existing experiments of Total S, ANC (Acid Neutralizing Capacity), and NAG respectively. To assure the applicability of the experiments, 36 samples from 19 sites with diverse geologies, field properties, and weathering conditions were collected. The samples were then subject to existing experiments and as a result, 14 samples which either were classified as UC or could be used as a comparison group had been selected. Afterwards, the selected samples were used to conduct the suggested

  5. CTEPP STANDARD OPERATING PROCEDURE FOR EXTRACTING AND PREPARING AIR SAMPLES FOR ANALYSIS OF POLAR PERSISTENT ORGANIC POLLUTANTS (SOP-5.13)

    EPA Science Inventory

    The method for extracting and preparing indoor and outdoor air samples for analysis of polar persistent organic pollutants is summarized in this SOP. It covers the preparation of samples that are to be analyzed by gas chromatography/mass spectrometry.

  6. Novel Sample Preparation Method for Safe and Rapid Detection of Bacillus anthracis Spores in Environmental Powders and Nasal Swabs

    PubMed Central

    Luna, Vicki A.; King, Debra; Davis, Carisa; Rycerz, Tony; Ewert, Matthew; Cannons, Andrew; Amuso, Philip; Cattani, Jacqueline

    2003-01-01

    Bacillus anthracis spores have been used as a biological weapon in the United States. We wanted to develop a safe, rapid method of sample preparation that provided safe DNA for the detection of spores in environmental and clinical specimens. Our method reproducibly detects B. anthracis in samples containing <10 spores. PMID:12624060

  7. Toward a Fieldable Atomic Mass Spectrometer for Safeguards Applications: Sample Preparation and Ionization

    SciTech Connect

    Barinaga, Charles J.; Hager, George J.; Hart, Garret L.; Koppenaal, David W.; Marcus, R. Kenneth; Jones, Sarah MH; Manard, Benjamin T.

    2014-10-31

    The International Atomic Energy Agency’s (IAEA’s) long-term research and development plan calls for the development of new methods to detect misuse at nuclear fuel cycle facilities such as reprocessing and enrichment plants. At enrichment plants, for example, the IAEA’s contemporary safeguards approaches are based on a combination of routine and random inspections that include collection of UF6 samples from in-process material and selected cylinders for subsequent analyses. These analyses include destructive analysis (DA) in a laboratory (typically by mass spectrometry [MS]) for isotopic characterization, and environmental sampling (ES) for subsequent laboratory elemental and isotopic analysis (also both typically by MS). One area of new method development includes moving this kind of isotope ratio analytical capability for DA and ES activities into the field. Some of the reasons for these developments include timeliness of results, avoidance of hazardous material shipments, and guidance for additional sample collecting. However, this capability does not already exist for several reasons, such as that most lab-based chemical and instrumental methods rely on laboratory infrastructure (highly trained staff, power, space, hazardous material handling, etc.) and require significant amounts of consumables (power, compressed gases, etc.). In addition, there are no currently available, fieldable instruments for atomic or isotope ratio analysis. To address these issues, Pacific Northwest National Laboratory (PNNL) and collaborator, Clemson University, are studying key areas that limit the fieldability of isotope ratio mass spectrometry for atomic ions: sample preparation and ionization, and reducing the physical size of a fieldable mass spectrometer. PNNL is seeking simple and robust techniques that could be effectively used by inspectors who may have no expertise in analytical MS. In this report, we present and describe the preliminary findings for three candidate

  8. Characterization and application of lactic acid bacteria for tropical silage preparation.

    PubMed

    Pholsen, Suradej; Khota, Waroon; Pang, Huili; Higgs, David; Cai, Yimin

    2016-10-01

    Strains TH 14, TH 21 and TH 64 were isolated from tropical silages, namely corn stover, sugar cane top and rice straw, respectively, prepared in Thailand. These strains were selected by low pH growth range and high lactic acid-producing ability, similar to some commercial inoculants. Based on the analysis of 16S ribosomal RNA gene sequence and DNA-DNA relatedness, strain TH 14 was identified as Lactobacillus casei, and strains TH 21 and TH 64 were identified as L. plantarum. Strains TH 14, TH 21, TH 64 and two commercial inoculants, CH (L. plantarum) and SN (L. rhamnosus), were used as additives to fresh and wilted purple Guinea and sorghum silages prepared using a small-scale fermentation method. The number of epiphytic lactic acid bacteria (LAB) in the forages before ensilage was relatively low but the numbers of coliform and aerobic bacteria were higher. Sorghum silages at 30 days of fermentation were all well preserved with low pH (3.56) and high lactic acid production (72.86 g/kg dry matter). Purple Guinea silage inoculated with LAB exhibited reduced count levels of aerobic and coliform bacteria, lower pH, butyric acid and ammonia nitrogen and increased lactic acid concentration, compared with the control. Strain TH 14 more effectively improved lactic acid production compared with inoculants and other strains. © 2016 Japanese Society of Animal Science.

  9. A sample preparation protocol for quantification of radiolabeled nucleoside incorporation into DNA by accelerator mass spectrometry

    NASA Astrophysics Data System (ADS)

    Hah, Sang Soo; Mundt, Janna M.; Ubick, Esther A.; Turteltaub, Kenneth W.; Gregg, Jeff P.; Henderson, Paul T.

    2007-06-01

    A general protocol is described for measuring the incorporation of radiocarbon-labeled 2‧-deoxynucleosides into DNA using accelerator mass spectrometry (AMS). This technology provides attomole (10-18 mol) sensitivity, with detection limits for DNA analysis in the range of one 14C atom per 1011-1012 total carbons. In practice this corresponds to approximately 1 labeled nucleoside per 1011 normal bases. A key aspect of the method is the use of precautions aimed at prevention of artifactual DNA oxidation during the sample preparation by the use of antioxidants and chaotropic salts during the DNA isolation. In principle, any type of appropriately labeled nucleoside derivative can be studied using the described protocol, provided that there is incorporation of the deoxynucleoside into DNA. We demonstrated this protocol using MCF-7 human breast cancer cells and a mouse model for mammary carcinoma, which we dosed with 14C-labeled 2‧-deoxyguanosine (dG) and 14C-labeled 7,8-dihydro-8-oxo-2‧-deoxyguanosine (8-oxodG). The nucleoside 8-oxodG is a ubiquitous compound that forms in cells by the reaction of dG with reactive oxygen species which has been associated with numerous disease, carcinogenesis and aging. DNA from cells treated with 14C-labeled nucleosides was isolated and prepared for analysis by AMS in order to measure the DNA-bound radioactivity. The method allows the generation of reliable and sufficient yields of pure DNA from human cells and animal tissues for analysis of radiocarbon levels. Ultimately, this protocol will be applied to understanding the role of modified nucleoside incorporation into DNA in cancer initiation and progression, but could also be used to study any DNA metabolism process where 14C-labeled nucleosides are used.

  10. Photo-oxidation of gaseous ethanol on photocatalyst prepared by acid leaching of titanium oxide/hydroxyapatite composite

    SciTech Connect

    Ono, Y.; Rachi, T.; Yokouchi, M.; Kamimoto, Y.; Nakajima, A.; Okada, K.

    2013-06-01

    Highlights: ► Photocatalyst powder was prepared by acid leaching of TiO{sub 2}/apatite composite. ► The photocatalytic activity was evaluated from in situ FT-IR study using ethanol. ► Apatite in the composite had positive effect for the photo-oxidation of ethanol. ► The enhanced oxidation rate was explained by the difference in deactivation rate. - Abstract: Highly active photocatalysts were synthesized by leaching of heat-treated titanium dioxide (TiO{sub 2})/hydroxyapatite (HAp) powder with hydrochloric acid at 0.25, 0.50, 0.75 mol/l, and their photocatalytic activities were evaluated from in situ Fourier transform infrared (FT-IR) study of photo-oxidation of gaseous ethanol. By changing the acid concentration, the TiO{sub 2}/HAp composite had different atomic ratios of Ca/Ti (0.0–2.8) and P/Ti (0.3–2.1). It was found that phosphate group remained on the surface of TiO{sub 2} particle even in the sample treated with concentrated acid (0.75 mol/l). These acid-treated samples showed higher rates for ethanol photo-oxidation than the commercial TiO{sub 2} powder, Degussa P25. The highest rate was obtained in the TiO{sub 2}/HAp composite treated with the dilute (0.25 mol/l) acid in spite of its low content of TiO{sub 2} photocatalyst. This enhanced photocatalytic activity was attributed to the result that the deactivation with repeated injections of ethanol gas was suppressed in the TiO{sub 2}/HAp composites compared with the TiO{sub 2} powders.

  11. Matrix-assisted laser desorption/ionization sample preparation optimization for structural characterization of poly(styrene-co-pentafluorostyrene) copolymers.

    PubMed

    Tisdale, Evgenia; Kennedy, Devin; Xu, Xiaodong; Wilkins, Charles

    2014-01-15

    The influence of the sample preparation parameters (the choice of the matrix, matrix:analyte ratio, salt:analyte ratio) was investigated and optimal conditions were established for the MALDI time-of-flight mass spectrometry analysis of the poly(styrene-co-pentafluorostyrene) copolymers. These were synthesized by atom transfer radical polymerization. Use of 2,5-dihydroxybenzoic acid as matrix resulted in spectra with consistently high ion yields for all matrix:analyte:salt ratios tested. The optimized MALDI procedure was successfully applied to the characterization of three copolymers obtained by varying the conditions of polymerization reaction. It was possible to establish the nature of the end groups, calculate molecular weight distributions, and determine the individual length distributions for styrene and pentafluorostyrene monomers, contained in the resulting copolymers. Based on the data obtained, it was concluded that individual styrene chain length distributions are more sensitive to the change in the composition of the catalyst (the addition of small amount of CuBr2) than is the pentafluorostyrene component distribution.

  12. Automated sample preparation techniques for the determination of drug enantiomers in biological fluids using liquid chromatography with chiral stationary phases.

    PubMed

    Ceccato, A; Toussaint, B; Chiap, P; Hubert, P; Crommen, J

    1999-01-01

    The determination of drug enantiomers has become of prime importance in the field of pharmaceutical and biomedical analysis. Liquid chromatography (LC) is one of the most frequently used techniques for achieving the separation and quantitation of the enantiomers of drug compounds. In the bioanalytical field, the integrated systems present an interesting alternative to time-consuming sample preparation techniques such as liquid-liquid extraction. Solid phase extraction (SPE) on disposable cartridges, dialysis or column switching are sample preparation techniques that can be fully automated and applied to enantioselective analysis in biological fluids. The selection of the most appropriate LC mode and chiral stationary phase for enantioseparations in bioanalysis is discussed and some aspects of these automated sample preparation procedures are compared, such as selectivity, detectability, elution of the analytes from the extraction sorbent, sample volume and analyte stability.

  13. Dynamic three-phase microextraction as a sample preparation technique prior to capillary electrophoresis.

    PubMed

    Hou, Li; Lee, Hian Kee

    2003-06-01

    Dynamic three-phase (liquid-liquid-liquid) microextraction was developed for capillary electrophoresis. Four aromatic amines as model compounds were extracted from 4-mL aqueous samples adjusted to basic condition (donor solution) through a small volume of organic solvent impregnated in a hollow fiber, which was held by the needle of a conventional syringe, and retracted into a 5-microL acidic acceptor solution inside the syringe. A renewable organic film and aqueous sample plug were formed inside the hollow fiber with the repeated movement of the syringe plunger enabled by a programmable syringe pump. This is believed to be the first reported instance of a semiautomated dynamic liquid-liquid-liquid microextraction (LLLME) procedure. Following this microextraction, the 5-microL acceptor solution was analyzed by capillary zone electrophoresis (CE). This new technique provided approximately 140-fold enrichment in 20 min. Utilizing 4-chloroaniline as internal standard, dynamic LLLME could provide good reproducibility (<4.0%). In addition, this method allowed the direct transfer of extracted analytes to a CE system for analysis.

  14. Pressurized liquid extraction as a sample preparation method for the analysis of isoflavones in pulses.

    PubMed

    Delgado-Zamarreño, María Milagros; Pérez-Martín, Lara; Bustamante-Rangel, Myriam; Carabias-Martínez, Rita

    2012-08-01

    In this work, we describe a rapid and simple analytical method that exploits pressurized liquid extraction (PLE) and liquid chromatography with diode array detection for the determination of isoflavones in samples of Spanish pulses. Confirmation of the analytes present was performed using ion-trap mass spectrometry. To optimize the PLE extraction, variables such as the dispersing agent, type of solvent and sample amount, and the experimental parameters, such as temperature and the number of extraction cycles, were studied. Separation was carried out using a reverse-phase C18 with polar endcapping as the stationary phase and acetonitrile/water with 0.2 % of formic acid, under a gradient regime, as the mobile phase. Optimal extraction of formononetin and biochanin-A from chickpeas with PLE was achieved using Hydromatrix as a dispersant agent, methanol/water (50:50), a temperature of 90 °C, and three cycles. The same optimal conditions-except methanol/water (75:25)-for solvent extraction were obtained for the extraction of daidzin, genistin, and formononetin from lentils. Recoveries ranged from 97 to 110 %, and standard deviations lower than 20 % were obtained. The contents obtained for daidzin in lentils using the proposed method were not significantly different from those obtained using another official method of analysis.

  15. Porous poly(D,L-lactic acid) foams with tunable structure and mechanical anisotropy prepared by supercritical carbon dioxide.

    PubMed

    Floren, Michael; Spilimbergo, Sara; Motta, Antonella; Migliaresi, Claudio

    2011-11-01

    The design and tunability of tissue scaffolds, such as pore size and geometry, is crucial to the success of an engineered tissue replacement. Moreover, the mechanical nature of a tissue scaffold should display properties similar to the tissue of interest; therefore, tunability of the foam mechanical properties is desirable. Polymeric foams prepared using supercritical carbon dioxide as a blowing agent has emerged in recent years as a promising technique to prepare porous scaffolds. While a number of groups have reported on the tailoring of scaffold morphologies by using gas foaming techniques, few have considered the effects of such processing conditions on the physical and mechanical anisotropy achieved. The aim of this study was to demonstrate the tunability of the structure and mechanical anisotropy of foams prepared using a variety of different gas foaming conditions. Porous poly(D,L lactic acid) foams were prepared by the systematic adjustment of processing conditions, namely pressure, temperature and venting time, resulting in an extensive range of scaffold morphologies. Characterization of sample anisotropy was achieved by mechanical evaluation of foam specimens both longitudinal and transverse to the foaming direction. The obtained mechanical properties demonstrated a strong dependence of the processing conditions on mechanical anisotropy and performance. Furthermore, results indicate that factors other than pore geometry may be necessary to define the mechanical behavior of the foam specimens. The favorable compressive moduli, coupled with large degrees of anisotropy, suggests these foams may have suitable application as scaffolds for bone tissue engineering.

  16. Preparation of Coated Valproic Acid and Sodium Valproate Sustained-release Matrix Tablets

    PubMed Central

    Phaechamud, T.; Mueannoom, W.; Tuntarawongsa, S.; Chitrattha, S.

    2010-01-01

    The aim of this research was to investigate the technique for preparation of coated valproic acid and sodium valproate sustained-release matrix tablets. Different diluents were tested and selected as the effective absorbent for oily valproic acid. Effect of the amount of absorbent and hydroxypropylmethylcellulose on drug release from valproic acid-sodium valproate matrix tablets prepared with wet granulation technique was evaluated in pH change system. Colloidal silicon dioxide effectively adsorbed liquid valproic acid during wet granulation and granule preparation. The amounts of colloidal silicon dioxide and hydroxypropylmethylcellulose employed in tablet formulations affected drug release from the tablets. The drug release was prominently sustained for over 12 h using hydroxypropylmethylcellulose-based hydrophilic matrix system. The mechanism of drug release through the matrix polymer was a diffusion control. The drug release profile of the developed matrix tablet was similar to Depakine Chrono®, providing the values of similarity factor (f2) and difference factor (f1) of 85.56 and 2.37, respectively. Eudragit® L 30 D-55 was used as effective subcoating material for core matrix tablets before over coating with hydroxypropylmethylcellulose film with organic base solvent. Drug release profile of coated matrix tablet was almost similar to that of Depakine Chrono®. PMID:20838520

  17. Effect of acid etching duration on tensile bond strength of composite resin bonded to erbium:yttrium-aluminium-garnet laser-prepared dentine. Preliminary study.

    PubMed

    Chousterman, M; Heysselaer, D; Dridi, S M; Bayet, F; Misset, B; Lamard, L; Peremans, A; Nyssen-Behets, C; Nammour, S

    2010-11-01

    The purpose of this study was to compare the tensile bond strength of composite resin bonded to erbium:yttrium-aluminium-garnet (Er:YAG) laser-prepared dentine after different durations of acid etching. The occlusal third of 68 human third molars was removed in order to expose the dentine surface. The teeth were randomly divided into five groups: group B (control group), prepared with bur and total etch system with 15 s acid etching [37% orthophosphoric acid (H(3)PO(4))]; group L15, laser photo-ablated dentine (200 mJ) (laser irradiation conditions: pulse duration 100 micros, air-water spray, fluence 31.45 J/ cm(2), 10 Hz, non-contact hand pieces, beam spot size 0.9 mm, irradiation speed 3 mm/s, and total irradiation time 2 x 40 s); group L30, laser prepared, laser conditioned and 30 s acid etching; group L60, laser prepared, laser conditioned and 60 s acid etching; group L90, laser prepared, laser conditioned and 90 s acid etching. A plot of composite resin was bonded onto each exposed dentine and then tested for tensile bond strength. The values obtained were statistically analysed by analysis of variance (ANOVA) coupled with the Tukey-Kramer test at the 95% level. A 90 s acid etching before bonding showed the best bonding value (P < 0.05) when compared with all the other groups including the control group. There is no significance difference between other groups, nor within each group and the control group. There was a significant increase in tensile bond strength of the samples acid etched for 90 s.

  18. Preparation of finely dispersed O/W emulsion from fatty acid solubilized in subcritical water.

    PubMed

    Khuwijitjaru, Pramote; Kimura, Yukitaka; Matsuno, Ryuichi; Adachi, Shuji

    2004-10-01

    A novel method for preparing a finely dispersed oil-in-water emulsion is proposed. Octanoic acid dissolved in water at a high temperature of 220 or 230 degrees C at 15 MPa was combined with an aqueous solution of a surfactant and then the mixture was cooled. When a nonionic surfactant, decaglycerol monolaurate (ML-750) or polyoxyethylene sorbitan monolaurate (Tween 20), was used, fine emulsions with a median oil droplet diameter of 100 nm or less were successfully prepared at ML-750 and Tween 20 concentrations of 0.083% (w/v) and 0.042%, respectively, or higher. The diameters were much smaller than those of oil droplets prepared by the conventional homogenization method using a rotor/stator homogenizer. However, an anionic surfactant, sodium dodecyl sulfate, was not adequate for the preparation of such fine emulsions by the proposed method. Although the interfacial tensions between octanoic acid and the surfactant solutions were measured at different temperatures, they were not an indication for selecting a surfactant for the successful preparation of the fine emulsion by the proposed method.

  19. Preparation of levoglucosenone through sulfuric acid promoted pyrolysis of bagasse at low temperature.

    PubMed

    Sui, Xian-wei; Wang, Zhi; Liao, Bing; Zhang, Ying; Guo, Qing-xiang

    2012-01-01

    Fast pyrolysis of bagasse pretreated by sulfuric acid was conducted in a fixed bed reactor to prepare levoglucosenone (LGO), a very important anhydrosugar for organic synthesis. The liquid yield and LGO yield were studied at temperatures from 240 to 350 °C and sulfuric acid loadings from 0.92 to 7.10 wt.%. An optimal LGO yield of 7.58 wt.% was obtained at 270 °C with a sulfuric acid pretreatment concentration of 0.05 M (corresponding to 4.28 wt.% sulfuric acid loading). For comparison, microcrystalline cellulose pretreated by 0.05 M sulfuric acid solution was pyrolyzed at temperature from 270 °C to 320 °C, and bagasse loaded with 3-5 wt.% phosphoric acid was pyrolyzed at temperature from 270 °C to 350 °C. The highest yield of LGO from bagasse was 30% higher than that from microcrystalline cellulose, and treatment with sulfuric acid allowed a 21% higher yield than treatment with phosphoric acid.

  20. Humidity-controlled preparation of frozen-hydrated biological samples for cryogenic coherent x-ray diffraction microscopy.

    PubMed

    Takayama, Yuki; Nakasako, Masayoshi

    2012-05-01

    Coherent x-ray diffraction microscopy (CXDM) has the potential to visualize the structures of micro- to sub-micrometer-sized biological particles, such as cells and organelles, at high resolution. Toward advancing structural studies on the functional states of such particles, here, we developed a system for the preparation of frozen-hydrated biological samples for cryogenic CXDM experiments. The system, which comprised a moist air generator, microscope, micro-injector mounted on a micromanipulator,