Sample records for laboratory extraction method

  1. An improved method for field extraction and laboratory analysis of large, intact soil cores

    USGS Publications Warehouse

    Tindall, J.A.; Hemmen, K.; Dowd, J.F.

    1992-01-01

    Various methods have been proposed for the extraction of large, undisturbed soil cores and for subsequent analysis of fluid movement within the cores. The major problems associated with these methods are expense, cumbersome field extraction, and inadequate simulation of unsaturated flow conditions. A field and laboratory procedure is presented that is economical, convenient, and simulates unsaturated and saturated flow without interface flow problems and can be used on a variety of soil types. In the field, a stainless steel core barrel is hydraulically pressed into the soil (30-cm diam. and 38 cm high), the barrel and core are extracted from the soil, and after the barrel is removed from the core, the core is then wrapped securely with flexible sheet metal and a stainless mesh screen is attached to the bottom of the core for support. In the laboratory the soil core is set atop a porous ceramic plate over which a soil-diatomaceous earth slurry has been poured to assure good contact between plate and core. A cardboard cylinder (mold) is fastened around the core and the empty space filled with paraffin wax. Soil cores were tested under saturated and unsaturated conditions using a hanging water column for potentials ???0. Breakthrough curves indicated that no interface flow occurred along the edge of the core. This procedure proved to be reliable for field extraction of large, intact soil cores and for laboratory analysis of solute transport.

  2. Extracting laboratory test information from biomedical text

    PubMed Central

    Kang, Yanna Shen; Kayaalp, Mehmet

    2013-01-01

    Background: No previous study reported the efficacy of current natural language processing (NLP) methods for extracting laboratory test information from narrative documents. This study investigates the pathology informatics question of how accurately such information can be extracted from text with the current tools and techniques, especially machine learning and symbolic NLP methods. The study data came from a text corpus maintained by the U.S. Food and Drug Administration, containing a rich set of information on laboratory tests and test devices. Methods: The authors developed a symbolic information extraction (SIE) system to extract device and test specific information about four types of laboratory test entities: Specimens, analytes, units of measures and detection limits. They compared the performance of SIE and three prominent machine learning based NLP systems, LingPipe, GATE and BANNER, each implementing a distinct supervised machine learning method, hidden Markov models, support vector machines and conditional random fields, respectively. Results: Machine learning systems recognized laboratory test entities with moderately high recall, but low precision rates. Their recall rates were relatively higher when the number of distinct entity values (e.g., the spectrum of specimens) was very limited or when lexical morphology of the entity was distinctive (as in units of measures), yet SIE outperformed them with statistically significant margins on extracting specimen, analyte and detection limit information in both precision and F-measure. Its high recall performance was statistically significant on analyte information extraction. Conclusions: Despite its shortcomings against machine learning methods, a well-tailored symbolic system may better discern relevancy among a pile of information of the same type and may outperform a machine learning system by tapping into lexically non-local contextual information such as the document structure. PMID:24083058

  3. A simple and fast method for extraction and quantification of cryptophyte phycoerythrin.

    PubMed

    Thoisen, Christina; Hansen, Benni Winding; Nielsen, Søren Laurentius

    2017-01-01

    The microalgal pigment phycoerythrin (PE) is of commercial interest as natural colorant in food and cosmetics, as well as fluoroprobes for laboratory analysis. Several methods for extraction and quantification of PE are available but they comprise typically various extraction buffers, repetitive freeze-thaw cycles and liquid nitrogen, making extraction procedures more complicated. A simple method for extraction of PE from cryptophytes is described using standard laboratory materials and equipment. The cryptophyte cells on the filters were disrupted at -80 °C and added phosphate buffer for extraction at 4 °C followed by absorbance measurement. The cryptophyte Rhodomonas salina was used as a model organism. •Simple method for extraction and quantification of phycoerythrin from cryptophytes.•Minimal usage of equipment and chemicals, and low labor costs.•Applicable for industrial and biological purposes.

  4. Natural colorants: Pigment stability and extraction yield enhancement via utilization of appropriate pretreatment and extraction methods.

    PubMed

    Ngamwonglumlert, Luxsika; Devahastin, Sakamon; Chiewchan, Naphaporn

    2017-10-13

    Natural colorants from plant-based materials have gained increasing popularity due to health consciousness of consumers. Among the many steps involved in the production of natural colorants, pigment extraction is one of the most important. Soxhlet extraction, maceration, and hydrodistillation are conventional methods that have been widely used in industry and laboratory for such a purpose. Recently, various non-conventional methods, such as supercritical fluid extraction, pressurized liquid extraction, microwave-assisted extraction, ultrasound-assisted extraction, pulsed-electric field extraction, and enzyme-assisted extraction have emerged as alternatives to conventional methods due to the advantages of the former in terms of smaller solvent consumption, shorter extraction time, and more environment-friendliness. Prior to the extraction step, pretreatment of plant materials to enhance the stability of natural pigments is another important step that must be carefully taken care of. In this paper, a comprehensive review of appropriate pretreatment and extraction methods for chlorophylls, carotenoids, betalains, and anthocyanins, which are major classes of plant pigments, is provided by using pigment stability and extraction yield as assessment criteria.

  5. A RAPID DNA EXTRACTION METHOD FOR PCR IDENTIFICATION OF FUNGAL INDOOR AIR CONTAMINANTS

    EPA Science Inventory

    Following air sampling, fungal DNA needs to be extracted and purified to a state suitable for laboratory use. Our laboratory has developed a simple method of extraction and purification of fungal DNA appropriate for enzymatic manipulation and polymerase chain reaction (PCR) appli...

  6. Interlaboratory comparison of extraction efficiency of pesticides from surface and laboratory water using solid-phase extraction disks.

    PubMed

    Senseman, Scott A; Mueller, Thomas C; Riley, Melissa B; Wauchope, R Don; Clegg, Chris; Young, Roddy W; Southwick, Lloyd M; Moye, H Anson; Dumas, Jose A; Mersie, Wondi; Mattice, John D; Leidy, Ross B

    2003-06-18

    A continuation of an earlier interlaboratory comparison was conducted (1) to assess solid-phase extraction (SPE) using Empore disks to extract atrazine, bromacil, metolachlor, and chlorpyrifos from various water sources accompanied by different sample shipping and quantitative techniques and (2) to compare quantitative results of individual laboratories with results of one common laboratory. Three replicates of a composite surface water (SW) sample were fortified with the analytes along with three replicates of deionized water (DW). A nonfortified DW sample and a nonfortified SW sample were also extracted. All samples were extracted using Empore C(18) disks. After extraction, part of the samples were eluted and analyzed in-house. Duplicate samples were evaporated in a 2-mL vial, shipped dry to a central laboratory (SDC), redissolved, and analyzed. Overall, samples analyzed in-house had higher recoveries than SDC samples. Laboratory x analysis type and laboratory x water source interactions were significant for all four compounds. Seven laboratories participated in this interlaboratory comparison program. No differences in atrazine recoveries were observed from in-house samples analyzed by laboratories A, B, D, and G compared with the recovery of SDC samples. In-house atrazine recoveries from laboratories C and F were higher when compared with recovery from SDC samples. However, laboratory E had lower recoveries from in-house samples compared with SDC samples. For each laboratory, lower recoveries were observed for chlorpyrifos from the SDC samples compared with samples analyzed in-house. Bromacil recovery was <65% at two of the seven laboratories in the study. Bromacil recoveries for the remaining laboratories were >75%. Three laboratories showed no differences in metolachlor recovery; two laboratories had higher recoveries for samples analyzed in-house, and two other laboratories showed higher metolachlor recovery for SDC samples. Laboratory G had a higher

  7. A 5-day method for determination of soluble silicon concentrations in nonliquid fertilizer materials using a sodium carbonate-ammonium nitrate extractant followed by visible spectroscopy with heteropoly blue analysis: single-laboratory validation.

    PubMed

    Sebastian, Dennis; Rodrigues, Hugh; Kinsey, Charles; Korndörfer, Gaspar; Pereira, Hamilton; Buck, Guilherme; Datnoff, Lawrence; Miranda, Stephen; Provance-Bowley, Mary

    2013-01-01

    A 5-day method for determining the soluble silicon (Si) concentrations in nonliquid fertilizer products was developed using a sodium carbonate (Na2CO3)-ammonium nitrate (NH4NO3) extractant followed by visible spectroscopy with heteropoly blue analysis at 660 nm. The 5-Day Na2CO3-NH4NO3 Soluble Si Extraction Method can be applied to quantify the plant-available Si in solid fertilizer products at levels ranging from 0.2 to 8.4% Si with an LOD of 0.06%, and LOQ of 0.20%. This Si extraction method for fertilizers correlates well with plant uptake of Si (r2 = 0.96 for a range of solid fertilizers) and is applicable to solid Si fertilizer products including blended products and beneficial substances. Fertilizer materials can be processed as received using commercially available laboratory chemicals and materials at ambient laboratory temperatures. The single-laboratory validation of the 5-Day Na2CO3-NH4NO3 Soluble Si Extraction Method has been approved by The Association of American Plant Food Control Officials for testing nonliquid Si fertilizer products.

  8. Project-Based Learning in Undergraduate Environmental Chemistry Laboratory: Using EPA Methods to Guide Student Method Development for Pesticide Quantitation

    ERIC Educational Resources Information Center

    Davis, Eric J.; Pauls, Steve; Dick, Jonathan

    2017-01-01

    Presented is a project-based learning (PBL) laboratory approach for an upper-division environmental chemistry or quantitative analysis course. In this work, a combined laboratory class of 11 environmental chemistry students developed a method based on published EPA methods for the extraction of dichlorodiphenyltrichloroethane (DDT) and its…

  9. Inorganic arsenic in seafood: does the extraction method matter?

    PubMed

    Pétursdóttir, Ásta H; Gunnlaugsdóttir, Helga; Krupp, Eva M; Feldmann, Jörg

    2014-05-01

    Nine different extraction methods were evaluated for three seafood samples to test whether the concentration of inorganic arsenic (iAs) determined in seafood is dependent on the extraction method. Certified reference materials (CRM) DOLT-4 (Dogfish Liver) and TORT-2 (Lobster Hepatopancreas), and a commercial herring fish meal were evaluated. All experimental work described here was carried out by the same operator using the same instrumentation, thus eliminating possible differences in results caused by laboratory related factors. Low concentrations of iAs were found in CRM DOLT-4 (0.012±0.003mgkg(-1)) and the herring fish meal sample (0.007±0.002mgkg(-1)) for all extraction methods. When comparing the concentration of iAs in CRM TORT-2 found in this study and in the literature dilute acids, HNO3 and HCl, showed the highest extracted iAs wheras dilute NaOH (in 50% ethanol) showed significantly lower extracted iAs. However, most other extraction solvents were not statistically different from one another. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Evaluation of a solid-phase extraction method for benzoylecgonine urine analysis in a high-throughput forensic urine drug-testing laboratory.

    PubMed

    Stout, Peter R; Gehlhausen, Jay M; Horn, Carl K; Klette, Kevin L

    2002-10-01

    A novel extraction and derivatization procedure for the cocaine metabolite benzoylecgonine (BZE) was developed and evaluated for use in a high-volume forensic urine analysis laboratory. Extractions utilized a Speedisk 48 positive pressure extraction manifold and polymer-based cation-exchange extraction columns. Samples were derivatized by the addition of pentafluoropropionic anhydride and pentafluoropropanol. All analyses were performed in selected ion monitoring mode; ions included m/z 421, 300, 272, 429, and 303 with m/z 421 to 429 ratio used for quantitation. The average extraction efficiency was 80%. Seventy-five common over-the-counter products, including prescription drugs, drug metabolites, and other drugs of abuse, demonstrated no significant interference with respect to chromatography or quantitation. The limit of detection and limit of quantitation were calculated at 12.5 ng/mL, and the assay was linear from 12.5 to 20,000 ng/mL with an r2 of 0.99932. A series of 20 precision samples (100 ng/mL) produced an average response of 97.8 ng/mL and a percent coefficient of variation of 4.1%. A set of 79 archived human urine samples that had previously been found to contain BZE were analyzed by 3 separate laboratories. The results did not differ significantly from prior quantitation or between laboratories. The Speedisk has proven viable for a high-volume production facility reducing overall cost of analysis by decreasing analysis time and minimizing waste production while meeting strict forensic requirements.

  11. Multi-laboratory survey of qPCR enterococci analysis method performance

    EPA Pesticide Factsheets

    Quantitative polymerase chain reaction (qPCR) has become a frequently used technique for quantifying enterococci in recreational surface waters, but there are several methodological options. Here we evaluated how three method permutations, type of mastermix, sample extract dilution and use of controls in results calculation, affect method reliability among multiple laboratories with respect to sample interference. Multiple samples from each of 22 sites representing an array of habitat types were analyzed using EPA Method 1611 and 1609 reagents with full strength and five-fold diluted extracts. The presence of interference was assessed three ways: using sample processing and PCR amplifications controls; consistency of results across extract dilutions; and relative recovery of target genes from spiked enterococci in water sample compared to control matrices with acceptable recovery defined as 50 to 200%. Method 1609, which is based on an environmental mastermix, was found to be superior to Method 1611, which is based on a universal mastermix. Method 1611 had over a 40% control assay failure rate with undiluted extracts and a 6% failure rate with diluted extracts. Method 1609 failed in only 11% and 3% of undiluted and diluted extracts analyses. Use of sample processing control assay results in the delta-delta Ct method for calculating relative target gene recoveries increased the number of acceptable recovery results. Delta-delta tended to bias recoveries fr

  12. Solid-phase extraction and GC-MS analysis of THC-COOH method optimized for a high-throughput forensic drug-testing laboratory.

    PubMed

    Stout, P R; Horn, C K; Klette, K L

    2001-10-01

    In order to facilitate the confirmation analysis of large numbers of urine samples previously screened positive for delta9-tetrahydrocannabinol (THC), an extraction, derivitization, and GC-MS analysis method was developed. This method utilized a positive pressure manifold anion-exchange polymer-based solid-phase extraction followed by elution directly into the automated liquid sampling (ALS) vials. Rapid derivitization was accomplished using pentafluoropropionic anhydride/pentafluoropropanol (PFPA/PFPOH). Recoveries averaged 95% with a limit of detection of 0.875 ng/mL with a 3-mL sample volume. Performance of 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH)-d3 and THC-COOH-d9 internal standards were evaluated. The method was linear to 900 ng/mL THC-COOH using THC-COOH-d9 with negligible contribution from the internal standard to very weak samples. Excellent agreement was seen with previous quantitations of human urine samples. More than 1000 human urine samples were analyzed using the method with 300 samples analyzed using an alternate qualifier ion (m/z 622) after some interference was observed with a qualifier ion (m/z 489). The 622 ion did not exhibit any interference even in samples with interfering peaks present in the 489 ion. The method resulted in dramatic reductions in processing time, waste production, and exposure hazards to laboratory personnel.

  13. Development, validation and evaluation of an analytical method for the determination of monomeric and oligomeric procyanidins in apple extracts.

    PubMed

    Hollands, Wendy J; Voorspoels, Stefan; Jacobs, Griet; Aaby, Kjersti; Meisland, Ane; Garcia-Villalba, Rocio; Tomas-Barberan, Francisco; Piskula, Mariusz K; Mawson, Deborah; Vovk, Irena; Needs, Paul W; Kroon, Paul A

    2017-04-28

    There is a lack of data for individual oligomeric procyanidins in apples and apple extracts. Our aim was to develop, validate and evaluate an analytical method for the separation, identification and quantification of monomeric and oligomeric flavanols in apple extracts. To achieve this, we prepared two types of flavanol extracts from freeze-dried apples; one was an epicatechin-rich extract containing ∼30% (w/w) monomeric (-)-epicatechin which also contained oligomeric procyanidins (Extract A), the second was an oligomeric procyanidin-rich extract depleted of epicatechin (Extract B). The parameters considered for method optimisation were HPLC columns and conditions, sample heating, mass of extract and dilution volumes. The performance characteristics considered for method validation included standard linearity, method sensitivity, precision and trueness. Eight laboratories participated in the method evaluation. Chromatographic separation of the analytes was best achieved utilizing a Hilic column with a binary mobile phase consisting of acidic acetonitrile and acidic aqueous methanol. The final method showed linearity for epicatechin in the range 5-100μg/mL with a correlation co-efficient >0.999. Intra-day and inter-day precision of the analytes ranged from 2 to 6% and 2 to 13% respectively. Up to dp3, trueness of the method was >95% but decreased with increasing dp. Within laboratory precision showed RSD values <5 and 10% for monomers and oligomers, respectively. Between laboratory precision was 4 and 15% (Extract A) and 7 and 30% (Extract B) for monomers and oligomers, respectively. An analytical method for the separation, identification and quantification of procyanidins in an apple extract was developed, validated and assessed. The results of the inter-laboratory evaluation indicate that the method is reliable and reproducible. Copyright © 2017. Published by Elsevier B.V.

  14. Use of Solid Phase Extraction in the Biochemistry Laboratory to Separate Different Lipids

    ERIC Educational Resources Information Center

    Flurkey, William H.

    2005-01-01

    Solid-phase extraction (SPE) was used to demonstrate how various lipids and lipid classes could be separated in a biochemistry laboratory setting. Three different SPE methods were chosen on their ability to separate a lipid mixture, consisting of a combination of a either a fatty acid, a triacylglycerol, a mono- or diacylglycerol, phospholipid,…

  15. Multi-Laboratory Study of Five Methods for the Determination of Brevetoxins in Shellfish Tissue Extracts.

    PubMed

    Dickey, Robert W; Plakas, Steven M; Jester, Edward L E; El Said, Kathleen R; Johannessen, Jan N; Flewelling, Leanne J; Scott, Paula; Hammond, Dan G; Van Dolah, Frances M; Leighfield, Tod A; Bottein Dachraoui, Marie-Yasmine; Ramsdell, John S; Pierce, Richard H; Henry, Mike S; Poli, Mark A; Walker, Calvin; Kurtz, Jan; Naar, Jerome; Baden, Daniel G; Musser, Steve M; White, Kevin D; Truman, Penelope; Miller, Aaron; Hawryluk, Timothy P; Wekell, Marleen M; Stirling, David; Quilliam, Michael A; Lee, Jung K

    A thirteen-laboratory comparative study tested the performance of four methods as alternatives to mouse bioassay for the determination of brevetoxins in shellfish. The methods were N2a neuroblastoma cell assay, two variations of the sodium channel receptor binding assay, competitive ELISA, and LC/MS. Three to five laboratories independently performed each method using centrally prepared spiked and naturally incurred test samples. Competitive ELISA and receptor binding (96-well format) compared most favorably with mouse bioassay. Between-laboratory relative standard deviations (RSDR) ranged from 10 to 20% for ELISA and 14 to 31% for receptor binding. Within-laboratory (RSDr) ranged from 6 to 15% for ELISA, and 5 to 31% for receptor binding. Cell assay was extremely sensitive but data variation rendered it unsuitable for statistical treatment. LC/MS performed as well as ELISA on spiked test samples but was inordinately affected by lack of toxin-metabolite standards, uniform instrumental parameters, or both, on incurred test samples. The ELISA and receptor binding assay are good alternatives to mouse bioassay for the determination of brevetoxins in shellfish.

  16. Designing easy DNA extraction: Teaching creativity through laboratory practice.

    PubMed

    Susantini, Endang; Lisdiana, Lisa; Isnawati; Tanzih Al Haq, Aushia; Trimulyono, Guntur

    2017-05-01

    Subject material concerning Deoxyribose Nucleic Acid (DNA) structure in the format of creativity-driven laboratory practice offers meaningful learning experience to the students. Therefore, a laboratory practice in which utilizes simple procedures and easy-safe-affordable household materials should be promoted to students to develop their creativity. This study aimed to examine whether designing and conducting DNA extraction with household materials could foster students' creative thinking. We also described how this laboratory practice affected students' knowledge and views. A total of 47 students participated in this study. These students were grouped and asked to utilize available household materials and modify procedures using hands-on worksheet. Result showed that this approach encouraged creative thinking as well as improved subject-related knowledge. Students also demonstrated positive views about content knowledge, social skills, and creative thinking skills. This study implies that extracting DNA with household materials is able to develop content knowledge, social skills, and creative thinking of the students. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(3):216-225, 2017. © 2016 The International Union of Biochemistry and Molecular Biology.

  17. Validation of a standardized extraction method for formalin-fixed paraffin-embedded tissue samples.

    PubMed

    Lagheden, Camilla; Eklund, Carina; Kleppe, Sara Nordqvist; Unger, Elizabeth R; Dillner, Joakim; Sundström, Karin

    2016-07-01

    Formalin-fixed paraffin-embedded (FFPE) samples can be DNA-extracted and used for human papillomavirus (HPV) genotyping. The xylene-based gold standard for extracting FFPE samples is laborious, suboptimal and involves health hazards for the personnel involved. To compare extraction with the standard xylene method to a xylene-free method used in an HPV LabNet Global Reference Laboratory at the Centers for Disease Control (CDC); based on a commercial method with an extra heating step. Fifty FFPE samples were randomly selected from a national audit of all cervical cancer cases diagnosed in Sweden during 10 years. For each case-block, a blank-block was sectioned, as a control for contamination. For xylene extraction, the standard WHO Laboratory Manual protocol was used. For the CDC method, the manufacturers' protocol was followed except for an extra heating step, 120°C for 20min. Samples were extracted and tested in parallel with β-globin real-time PCR, HPV16 real-time PCR and HPV typing using modified general primers (MGP)-PCR and Luminex assays. For a valid result the blank-block had to be betaglobin-negative in all tests and the case-block positive for beta-globin. Overall, detection was improved with the heating method and the amount of HPV-positive samples increased from 70% to 86% (p=0.039). For all samples where HPV type concordance could be evaluated, there was 100% type concordance. A xylene-free and robust extraction method for HPV-DNA typing in FFPE material is currently in great demand. Our proposed standardized protocol appears to be generally useful. Copyright © 2016. Published by Elsevier B.V.

  18. An Improved Method for Extraction and Separation of Photosynthetic Pigments

    ERIC Educational Resources Information Center

    Katayama, Nobuyasu; Kanaizuka, Yasuhiro; Sudarmi, Rini; Yokohama, Yasutsugu

    2003-01-01

    The method for extracting and separating hydrophobic photosynthetic pigments proposed by Katayama "et al." ("Japanese Journal of Phycology," 42, 71-77, 1994) has been improved to introduce it to student laboratories at the senior high school level. Silica gel powder was used for removing water from fresh materials prior to…

  19. Liquid-Liquid Extraction of Insecticides from Juice: An Analytical Chemistry Laboratory Experiment

    ERIC Educational Resources Information Center

    Radford, Samantha A.; Hunter, Ronald E., Jr.; Barr, Dana Boyd; Ryan, P. Barry

    2013-01-01

    A laboratory experiment was developed to target analytical chemistry students and to teach them about insecticides in food, sample extraction, and cleanup. Micro concentrations (sub-microgram/mL levels) of 12 insecticides spiked into apple juice samples are extracted using liquid-liquid extraction and cleaned up using either a primary-secondary…

  20. Evaluation of micro-colorimetric lipid determination method with samples prepared using sonication and accelerated solvent extraction methods

    EPA Science Inventory

    Two common laboratory extraction techniques were evaluated for routine use with the micro-colorimetric lipid determination method developed by Van Handel (1985) [E. Van Handel, J. Am. Mosq. Control Assoc. 1(1985) 302] and recently validated for small samples by Inouye and Lotufo ...

  1. Filtration Isolation of Nucleic Acids: A Simple and Rapid DNA Extraction Method.

    PubMed

    McFall, Sally M; Neto, Mário F; Reed, Jennifer L; Wagner, Robin L

    2016-08-06

    FINA, filtration isolation of nucleic acids, is a novel extraction method which utilizes vertical filtration via a separation membrane and absorbent pad to extract cellular DNA from whole blood in less than 2 min. The blood specimen is treated with detergent, mixed briefly and applied by pipet to the separation membrane. The lysate wicks into the blotting pad due to capillary action, capturing the genomic DNA on the surface of the separation membrane. The extracted DNA is retained on the membrane during a simple wash step wherein PCR inhibitors are wicked into the absorbent blotting pad. The membrane containing the entrapped DNA is then added to the PCR reaction without further purification. This simple method does not require laboratory equipment and can be easily implemented with inexpensive laboratory supplies. Here we describe a protocol for highly sensitive detection and quantitation of HIV-1 proviral DNA from 100 µl whole blood as a model for early infant diagnosis of HIV that could readily be adapted to other genetic targets.

  2. A Guided Inquiry Liquid/Liquid Extractions Laboratory for Introductory Organic Chemistry

    ERIC Educational Resources Information Center

    Raydo, Margaret L.; Church, Megan S.; Taylor, Zane W.; Taylor, Christopher E.; Danowitz, Amy M.

    2015-01-01

    A guided inquiry laboratory experiment for teaching liquid/liquid extractions to first semester undergraduate organic chemistry students is described. This laboratory is particularly useful for introductory students as the analytes that are separated are highly colored dye molecules. This allows students to track into which phase each analyte…

  3. Biotechnology Laboratory Methods.

    ERIC Educational Resources Information Center

    Davis, Robert H.; Kompala, Dhinakar S.

    1989-01-01

    Describes a course entitled "Biotechnology Laboratory" which introduces a variety of laboratory methods associated with biotechnology. Describes the history, content, and seven experiments of the course. The seven experiments are selected from microbiology and molecular biology, kinetics and fermentation, and downstream…

  4. Analysis of laboratory compaction methods of roller compacted concrete

    NASA Astrophysics Data System (ADS)

    Trtík, Tomáš; Chylík, Roman; Bílý, Petr; Fládr, Josef

    2017-09-01

    Roller-Compacted Concrete (RCC) is an ordinary concrete poured and compacted with machines typically used for laying of asphalt road layers. One of the problems connected with this technology is preparation of representative samples in the laboratory. The aim of this work was to analyse two methods of preparation of RCC laboratory samples with bulk density as the comparative parameter. The first method used dynamic compaction by pneumatic hammer. The second method of compaction had a static character. The specimens were loaded by precisely defined force in laboratory loading machine to create the same conditions as during static rolling (in the Czech Republic, only static rolling is commonly used). Bulk densities obtained by the two compaction methods were compared with core drills extracted from real RCC structure. The results have shown that the samples produced by pneumatic hammer tend to overestimate the bulk density of the material. For both compaction methods, immediate bearing index test was performed to verify the quality of compaction. A fundamental difference between static and dynamic compaction was identified. In static compaction, initial resistance to penetration of the mandrel was higher, after exceeding certain limit the resistance was constant. This means that the samples were well compacted just on the surface. Specimens made by pneumatic hammer actively resisted throughout the test, the whole volume was uniformly compacted.

  5. ON-SITE SOLID-PHASE EXTRACTION AND LABORATORY ...

    EPA Pesticide Factsheets

    Fragrance materials such as synthetic musks in aqueous samples, are normally determined by gas chromatography/mass spectrometry in the selected ion monitoring (SIM) mode to provide maximum sensitivity after liquid-liquid extraction of I -L samples. Full-scan mass spectra are required to verify that a target analyte has been found by comparison with the mass spectra of fragrance compounds in the NIST mass spectral library. A I -L sample usually provides insufficient analyte for full scan data acquisition. This paper describes an on-site extraction method developed at the U.S. Environmental Protection Agency (USEPA)- Las Vegas Nevada - for synthetic musks from 60 L of wastewater effluent. Such a large sample volume permits high-quality, full-scan mass spectra to be obtained for a wide array of synthetic musks. Quantification of these compounds was achieved from the full-scan data directly, without the need to acquire SIM data. The detection limits obtained with this method are an order of magnitude lower than those obtained from liquid-liquid and other solid phase extraction methods. This method is highly reproducible, and recoveries ranged from 80 to 97% in spiked sewage treatment plant effluent. The high rate of sorbent-sample mass transfer eliminated the need for a methanolic activation step, which reduced extraction time, labor, and solvent use, More samples could be extracted in the field at lower cost. After swnple extraction, the light- weight cartridges ar

  6. Flotation of Mineral and Dyes: A Laboratory Experiment for Separation Method Molecular Hitchhikers

    ERIC Educational Resources Information Center

    Rappon, Tim; Sylvestre, Jarrett A.; Rappon, Manit

    2016-01-01

    Flotation as a method of separation is widely researched and is applied in many industries. It has been used to address a wide range of environmental issues including treatment of wastewater, recovery of heavy metals for recycling, extraction of minerals in mining, and so forth. This laboratory attempts to show how such a simple method can be used…

  7. Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory; determination of pesticides in water by Carbopak-B solid-phase extraction and high-preformance liquid chromatography

    USGS Publications Warehouse

    Werner, Stephen L.; Burkhardt, Mark R.; DeRusseau, Sabrina N.

    1996-01-01

    In accordance with the needs of the National Water-Quality Assessment Program (NAWQA), the U.S. Geological Survey has developed and implemented a graphitized carbon-based solid-phase extraction and high-performance liquid chromatographic analytical method. The method is used to determine 41 pesticides and pesticide metabolites that are not readily amenable to gas chromatography or other high-temperature analytical techniques. Pesticides are extracted from filtered environmental water samples using a 0.5-gram graphitized carbon-based solid-phase cartridge, eluted from the cartridge into two analytical fractions, and analyzed using high-performance liquid chromatography with photodiode-array detection. The upper concentration limit is 1.6 micrograms per liter (=B5g/L) for most compounds. Single-operator method detection limits in organic-free water samples ranged from 0.006 to 0.032 =B5g/L= Recoveries in organic-free water samples ranged from 37 to 88 percent. Recoveries in ground- and surface-water samples ranged from 29 to 94 percent. An optional on-site extraction procedure allows for samples to be collected and processed at remote sites where it is difficult to ship samples to the laboratory within the recommended pre-extraction holding time of 7 days.

  8. Evaluation of methods for the extraction of DNA from drinking water distribution system biofilms.

    PubMed

    Hwang, Chiachi; Ling, Fangqiong; Andersen, Gary L; LeChevallier, Mark W; Liu, Wen-Tso

    2012-01-01

    While drinking water biofilms have been characterized in various drinking water distribution systems (DWDS), little is known about the impact of different DNA extraction methods on the subsequent analysis of microbial communities in drinking water biofilms. Since different DNA extraction methods have been shown to affect the outcome of microbial community analysis in other environments, it is necessary to select a DNA extraction method prior to the application of molecular tools to characterize the complex microbial ecology of the DWDS. This study compared the quantity and quality of DNA yields from selected DWDS bacteria with different cell wall properties using five widely used DNA extraction methods. These were further selected and evaluated for their efficiency and reproducibility of DNA extraction from DWDS samples. Terminal restriction fragment length analysis and the 454 pyrosequencing technique were used to interpret the differences in microbial community structure and composition, respectively, from extracted DNA. Such assessments serve as a concrete step towards the determination of an optimal DNA extraction method for drinking water biofilms, which can then provide a reliable comparison of the meta-analysis results obtained in different laboratories.

  9. Evaluation of Methods for the Extraction of DNA from Drinking Water Distribution System Biofilms

    PubMed Central

    Hwang, Chiachi; Ling, Fangqiong; Andersen, Gary L.; LeChevallier, Mark W.; Liu, Wen-Tso

    2012-01-01

    While drinking water biofilms have been characterized in various drinking water distribution systems (DWDS), little is known about the impact of different DNA extraction methods on the subsequent analysis of microbial communities in drinking water biofilms. Since different DNA extraction methods have been shown to affect the outcome of microbial community analysis in other environments, it is necessary to select a DNA extraction method prior to the application of molecular tools to characterize the complex microbial ecology of the DWDS. This study compared the quantity and quality of DNA yields from selected DWDS bacteria with different cell wall properties using five widely used DNA extraction methods. These were further selected and evaluated for their efficiency and reproducibility of DNA extraction from DWDS samples. Terminal restriction fragment length analysis and the 454 pyrosequencing technique were used to interpret the differences in microbial community structure and composition, respectively, from extracted DNA. Such assessments serve as a concrete step towards the determination of an optimal DNA extraction method for drinking water biofilms, which can then provide a reliable comparison of the meta-analysis results obtained in different laboratories. PMID:22075624

  10. Apparatus and methods for hydrocarbon extraction

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

    Bohnert, George W.; Verhulst, Galen G.

    Systems and methods for hydrocarbon extraction from hydrocarbon-containing material. Such systems and methods relate to extracting hydrocarbon from hydrocarbon-containing material employing a non-aqueous extractant. Additionally, such systems and methods relate to recovering and reusing non-aqueous extractant employed for extracting hydrocarbon from hydrocarbon-containing material.

  11. Supercritical Fluid Extraction versus Traditional Solvent Extraction of Caffeine from Tea Leaves: A Laboratory-Based Case Study for an Organic Chemistry Course

    ERIC Educational Resources Information Center

    Schaber, Peter M.; Larkin, Judith E.; Pines, Harvey A.; Berchou, Kelly; Wierchowski, Elizabeth; Marconi, Andrew; Suriani, Allison

    2012-01-01

    In this case-based laboratory, an instrument sales person attempts to convince an analysis laboratory of the virtues of supercritical fluid extraction (SFE). The sales person deals directly with the laboratory technicians who will make the decision. Arrangements are made to have SFE instrumentation brought into the laboratory for a comparative…

  12. Evaluating the efficacy of DNA differential extraction methods for sexual assault evidence.

    PubMed

    Klein, Sonja B; Buoncristiani, Martin R

    2017-07-01

    Analysis of sexual assault evidence, often a mixture of spermatozoa and victim epithelial cells, represents a significant portion of a forensic DNA laboratory's case load. Successful genotyping of sperm DNA from these mixed cell samples, particularly with low amounts of sperm, depends on maximizing sperm DNA recovery and minimizing non-sperm DNA carryover. For evaluating the efficacy of the differential extraction, we present a method which uses a Separation Potential Ratio (SPRED) to consider both sperm DNA recovery and non-sperm DNA removal as variables for determining separation efficiency. In addition, we describe how the ratio of male-to-female DNA in the sperm fraction may be estimated by using the SPRED of the differential extraction method in conjunction with the estimated ratio of male-to-female DNA initially present on the mixed swab. This approach may be useful for evaluating or modifying differential extraction methods, as we demonstrate by comparing experimental results obtained from the traditional differential extraction and the Erase Sperm Isolation Kit (PTC © ) procedures. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Simple, miniaturized blood plasma extraction method.

    PubMed

    Kim, Jin-Hee; Woenker, Timothy; Adamec, Jiri; Regnier, Fred E

    2013-12-03

    A rapid plasma extraction technology that collects a 2.5 μL aliquot of plasma within three minutes from a finger-stick derived drop of blood was evaluated. The utility of the plasma extraction cards used was that a paper collection disc bearing plasma was produced that could be air-dried in fifteen minutes and placed in a mailing envelop for transport to an analytical laboratory. This circumvents the need for venipuncture and blood collection in specialized vials by a phlebotomist along with centrifugation and refrigerated storage. Plasma extraction was achieved by applying a blood drop to a membrane stack through which plasma was drawn by capillary action. During the course of plasma migration to a collection disc at the bottom of the membrane stack blood cells were removed by a combination of adsorption and filtration. After the collection disc filled with an aliquot of plasma the upper membranes were stripped from the collection card and the collection disc was air-dried. Intercard differences in the volume of plasma collected varied approximately 1% while volume variations of less than 2% were seen with hematocrit levels ranging from 20% to 71%. Dried samples bearing metabolites and proteins were then extracted from the disc and analyzed. 25-Hydroxy vitamin D was quantified by LC-MS/MS analysis following derivatization with a secosteroid signal enhancing tag that imparted a permanent positive charge to the vitamin and reduced the limit of quantification (LOQ) to 1 pg of collected vitamin on the disc; comparable to values observed with liquid-liquid extraction (LLE) of a venipuncture sample. A similar study using conventional proteomics methods and spectral counting for quantification was conducted with yeast enolase added to serum as an internal standard. The LOQ with extracted serum samples for enolase was 1 μM, linear from 1 to 40 μM, the highest concentration examined. In all respects protein quantification with extracted serum samples was comparable to

  14. Comparison of methods of extracting messenger Ribonucleic Acid from ejaculated Porcine (Sus Scrofa) Spermatozoa

    USDA-ARS?s Scientific Manuscript database

    H. D. Guthrie, G.R. Welch, and L. A. Blomberg. Comparison of Methods of Extracting Messenger Ribonucleic Acid from Ejaculated Porcine (Sus Scrofa) Spermatozoa. Biotechnology and Germplasm Laboratory, Agricultural Research Service U. S. Department of Agriculture, Beltsville, MD 20705 The purpos...

  15. ACCELERATED SOLVENT EXTRACTION COMBINED WITH ...

    EPA Pesticide Factsheets

    A research project was initiated to address a recurring problem of elevated detection limits above required risk-based concentrations for the determination of semivolatile organic compounds in high moisture content solid samples. This project was initiated, in cooperation with the EPA Region 1 Laboratory, under the Regional Methods Program administered through the ORD Office of Science Policy. The aim of the project was to develop an approach for the rapid removal of water in high moisture content solids (e.g., wetland sediments) in preparation for analysis via Method 8270. Alternative methods for water removal have been investigated to enhance compound solid concentrations and improve extraction efficiency, with the use of pressure filtration providing a high-throughput alternative for removal of the majority of free water in sediments and sludges. In order to eliminate problems with phase separation during extraction of solids using Accelerated Solvent Extraction, a variation of a water-isopropanol extraction method developed at the USGS National Water Quality Laboratory in Denver, CO is being employed. The concentrations of target compounds in water-isopropanol extraction fluids are subsequently analyzed using an automated Solid Phase Extraction (SPE)-GC/MS method developed in our laboratory. The coupled approaches for dewatering, extraction, and target compound identification-quantitation provide a useful alternative to enhance sample throughput for Me

  16. Method of infusion extraction

    NASA Technical Reports Server (NTRS)

    Chang-Diaz, Franklin R. (Inventor)

    1989-01-01

    Apparatus and method of removing desirable constituents from an infusible material by infusion extraction, where a piston operating in a first chamber draws a solvent into the first chamber where it may be heated, and then moves the heated solvent into a second chamber containing the infusible material, and where infusion extraction takes place. The piston then moves the solvent containing the extract through a filter into the first chamber, leaving the extraction residue in the second chamber.

  17. Development of a modified cortisol extraction procedure for intermediately sized fish not amenable to whole-body or plasma extraction methods.

    PubMed

    Guest, Taylor W; Blaylock, Reginald B; Evans, Andrew N

    2016-02-01

    The corticosteroid hormone cortisol is the central mediator of the teleost stress response. Therefore, the accurate quantification of cortisol in teleost fishes is a vital tool for addressing fundamental questions about an animal's physiological response to environmental stressors. Conventional steroid extraction methods using plasma or whole-body homogenates, however, are inefficient within an intermediate size range of fish that are too small for phlebotomy and too large for whole-body steroid extractions. To assess the potential effects of hatchery-induced stress on survival of fingerling hatchery-reared Spotted Seatrout (Cynoscion nebulosus), we developed a novel extraction procedure for measuring cortisol in intermediately sized fish (50-100 mm in length) that are not amenable to standard cortisol extraction methods. By excising a standardized portion of the caudal peduncle, this tissue extraction procedure allows for a small portion of a larger fish to be sampled for cortisol, while minimizing the potential interference from lipids that may be extracted using whole-body homogenization procedures. Assay precision was comparable to published plasma and whole-body extraction procedures, and cortisol quantification over a wide range of sample dilutions displayed parallelism versus assay standards. Intra-assay %CV was 8.54%, and average recovery of spiked samples was 102%. Also, tissue cortisol levels quantified using this method increase 30 min after handling stress and are significantly correlated with blood values. We conclude that this modified cortisol extraction procedure provides an excellent alternative to plasma and whole-body extraction procedures for intermediately sized fish, and will facilitate the efficient assessment of cortisol in a variety of situations ranging from basic laboratory research to industrial and field-based environmental health applications.

  18. Application of the BioMek 2000 Laboratory Automation Workstation and the DNA IQ System to the extraction of forensic casework samples.

    PubMed

    Greenspoon, Susan A; Ban, Jeffrey D; Sykes, Karen; Ballard, Elizabeth J; Edler, Shelley S; Baisden, Melissa; Covington, Brian L

    2004-01-01

    Robotic systems are commonly utilized for the extraction of database samples. However, the application of robotic extraction to forensic casework samples is a more daunting task. Such a system must be versatile enough to accommodate a wide range of samples that may contain greatly varying amounts of DNA, but it must also pose no more risk of contamination than the manual DNA extraction methods. This study demonstrates that the BioMek 2000 Laboratory Automation Workstation, used in combination with the DNA IQ System, is versatile enough to accommodate the wide range of samples typically encountered by a crime laboratory. The use of a silica coated paramagnetic resin, as with the DNA IQ System, facilitates the adaptation of an open well, hands off, robotic system to the extraction of casework samples since no filtration or centrifugation steps are needed. Moreover, the DNA remains tightly coupled to the silica coated paramagnetic resin for the entire process until the elution step. A short pre-extraction incubation step is necessary prior to loading samples onto the robot and it is at this step that most modifications are made to accommodate the different sample types and substrates commonly encountered with forensic evidentiary samples. Sexual assault (mixed stain) samples, cigarette butts, blood stains, buccal swabs, and various tissue samples were successfully extracted with the BioMek 2000 Laboratory Automation Workstation and the DNA IQ System, with no evidence of contamination throughout the extensive validation studies reported here.

  19. Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory - Determination of Moderate-Use Pesticides and Selected Degradates in Water by C-18 Solid-Phase Extraction and Gas Chromatography/Mass Spectrometry

    USGS Publications Warehouse

    Sandstrom, Mark W.; Stroppel, Max E.; Foreman, William T.; Schroeder, Michael P.

    2001-01-01

    A method for the isolation and analysis of 21 parent pesticides and 20 pesticide degradates in natural-water samples is described. Water samples are filtered to remove suspended particulate matter and then are pumped through disposable solid-phase-extraction columns that contain octadecyl-bonded porous silica to extract the analytes. The columns are dried by using nitrogen gas, and adsorbed analytes are eluted with ethyl acetate. Extracted analytes are determined by capillary-column gas chromatography/mass spectrometry with selected-ion monitoring of three characteristic ions. The upper concentration limit is 2 micrograms per liter (?g/L) for most analytes. Single-operator method detection limits in reagent-water samples range from 0.00 1 to 0.057 ?g/L. Validation data also are presented for 14 parent pesticides and 20 degradates that were determined to have greater bias or variability, or shorter holding times than the other compounds. The estimated maximum holding time for analytes in pesticide-grade water before extraction was 4 days. The estimated maximum holding time for analytes after extraction on the dry solid-phase-extraction columns was 7 days. An optional on-site extraction procedure allows for samples to be collected and processed at remote sites where it is difficult to ship samples to the laboratory within the recommended pre-extraction holding time. The method complements existing U.S. Geological Survey Method O-1126-95 (NWQL Schedules 2001 and 2010) by using identical sample preparation and comparable instrument analytical conditions so that sample extracts can be analyzed by either method to expand the range of analytes determined from one water sample.

  20. Materials and Methods for Streamlined Laboratory Analysis of Environmental Samples, FY 2016 Report

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

    Addleman, Raymond S.; Naes, Benjamin E.; McNamara, Bruce K.

    The International Atomic Energy Agency (IAEA) relies upon laboratory analysis of environmental samples (typically referred to as “swipes”) collected during on-site inspections of safeguarded facilities to support the detection and deterrence of undeclared activities. Unfortunately, chemical processing and assay of the samples is slow and expensive. A rapid, effective, and simple extraction process and analysis method is needed to provide certified results with improved timeliness at reduced costs (principally in the form of reduced labor), while maintaining or improving sensitivity and efficacy. To address these safeguard needs the Pacific Northwest National Laboratory (PNNL) explored and demonstrated improved methods for environmentalmore » sample (ES) analysis. Improvements for both bulk and particle analysis were explored. To facilitate continuity and adoption, the new sampling materials and processing methods will be compatible with existing IAEA protocols for ES analysis. PNNL collaborated with Oak Ridge National Laboratory (ORNL), which performed independent validation of the new bulk analysis methods and compared performance to traditional IAEA’s Network of Analytical Laboratories (NWAL) protocol. ORNL efforts are reported separately. This report describes PNNL’s FY 2016 progress, which was focused on analytical application supporting environmental monitoring of uranium enrichment plants and nuclear fuel processing. In the future the technology could be applied to other safeguard applications and analytes related to fuel manufacturing, reprocessing, etc. PNNL’s FY 2016 efforts were broken into two tasks and a summary of progress, accomplishments and highlights are provided below. Principal progress and accomplishments on Task 1, Optimize Materials and Methods for ICP-MS Environmental Sample Analysis, are listed below. • Completed initial procedure for rapid uranium extraction from ES swipes based upon carbonate-peroxide chemistry (delivered to

  1. Extraction Methods in Soil Phosphorus Characterisation

    NASA Astrophysics Data System (ADS)

    Soinne, Helena

    2010-05-01

    Extraction methods are widely used to assess the bioavailability of P and to characterise soil P reserves. Even though new and more sophisticated methods to characterise soil P are constantly developed the use of extraction methods is not likely to be replaced because of the relatively simple analytical equipment needed for the analysis. However, the large variety of extractants, pre-treatments and sample preparation procedures complicate the comparison of published results. In order to improve our understanding of the behaviour and cycling of P in soil, it is important to know the role of extracted P in the soil P cycle. The knowledge of the factors affecting the analytical outcome is a prerequisite for justified interpretation of the results. In this study, the effect of sample pre-treatment and properties of the used extractant on extractable molybdate-reactive phosphorus (MRP) and molybdate-unreactive phosphorus (MUP) was studied. Furthermore, the effect of sample preparation procedures prior the analysis on measured MRP and MUP was studied. Two widely used sequential extraction procedures were compared on their ability to show management induced differences on soil P. These results revealed that pre-treatments changed soil properties and air-drying was found to affect soil P, particularly extractable MUP, thought to represent organic P, by disrupting organic matter. This was evidenced by an increase in the water-extractable small-sized (<0.2 µm) P that, at least partly, took place at the expense of the large-sized (>0.2 µm) P. In addition to the effects of sample pre-treatment, the results showed that extractable organic P was sensitive to the chemical nature of the used extractant and to the sample preparation procedures employed prior to P analysis, including centrifugation and filtering of soil suspensions. Filtering may remove a major proportion of extractable MUP; therefore filtering cannot be recommended in the characterisation of solubilised MUP

  2. Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa.

    PubMed

    Siegel, Chloe S; Stevenson, Florence O; Zimmer, Elizabeth A

    2017-02-01

    An efficient, effective DNA extraction method is necessary for comprehensive analysis of plant genomes. This study analyzed the quality of DNA obtained using paper FTA cards prepared directly in the field when compared to the more traditional cetyltrimethylammonium bromide (CTAB)-based extraction methods from silica-dried samples. DNA was extracted using FTA cards according to the manufacturer's protocol. In parallel, CTAB-based extractions were done using the automated AutoGen DNA isolation system. DNA quality for both methods was determined for 15 non-agricultural species collected in situ, by gel separation, spectrophotometry, fluorometry, and successful amplification and sequencing of nuclear and chloroplast gene markers. The FTA card extraction method yielded less concentrated, but also less fragmented samples than the CTAB-based technique. The card-extracted samples provided DNA that could be successfully amplified and sequenced. The FTA cards are also useful because the collected samples do not require refrigeration, extensive laboratory expertise, or as many hazardous chemicals as extractions using the CTAB-based technique. The relative success of the FTA card method in our study suggested that this method could be a valuable tool for studies in plant population genetics and conservation biology that may involve screening of hundreds of individual plants. The FTA cards, like the silica gel samples, do not contain plant material capable of propagation, and therefore do not require permits from the U.S. Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS) for transportation.

  3. SANDIA NATIONAL LABORATORIES IN SITU ELECTROKINETIC EXTRACTION TECHNOLOGY; INNOVATIVE TECHNOLOGY EVALUATION REPORT

    EPA Science Inventory

    As a part of the Superfund Innovative Technology Evaluation (SITE) Program, the U.S. Environmental Protection Agency evaluated the In-Situ Electrokinetic Extraction (ISEE) system at Sandia National Laboratories, Albuquerque, New Mexico.

    The SITE demonstration results show ...

  4. Evaluation of extraction methods for hexavalent chromium determination in dusts, ashes, and soils

    USGS Publications Warehouse

    Wolf, Ruth E.; Wilson, Stephen A.

    2010-01-01

    One of the difficulties in performing speciation analyses on solid samples is finding a suitable extraction method. Traditional methods for extraction of hexavalent chromium, Cr(VI), in soils, such as SW846 Method 3060A, can be tedious and are not always compatible with some determination methods. For example, the phosphate and high levels of carbonate and magnesium present in the U.S. Environmental Protection Agency (USEPA) Method 3060A digestion for Cr(VI) were found to be incompatible with the High Performance Liquid Chromatography-Inductively Coupled Plasma-Mass Spectrometry (HPLC-ICP-MS) detection method used by our laboratory. Modification of Method 3060A by eliminating the use of the phosphate buffer provided improved performance with the detection method, however dilutions are still necessary to achieve good chromatographic separation and detection of Cr(VI). An ultrasonic extraction method using a 1 mM Na2CO3 - 9 mM NaHCO3 buffer solution, adapted from Occupational Safety and Health Administration (OSHA) Method ID215, has been used with good results for the determination of Cr(VI) in air filters. The average recovery obtained for BCR-545 - Welding Dust Loaded on Filter (IRMM, Belgium) using this method was 99 percent (1.2 percent relative standard deviation) with no conversion of Cr(VI) to Cr(III) during the extraction process. This ultrasonic method has the potential for use with other sample matrices, such as ashes and soils. Preliminary investigations using NIST 2701 (Hexavalent Chromium in Contaminated Soil) loaded onto quartz filters showed promising results with approximately 90 percent recovery of the certified Cr(VI) value. Additional testing has been done using NIST 2701 and NIST 2700 using different presentation methods. Extraction efficiency of bulk presentation, where small portions of the sample are added to the bottom of the extraction vessel, will be compared with supported presentation, where small portions of the sample are loaded onto a

  5. Linking Laboratory Experiences to the Real World: The Extraction of Octylphenoxyacetic Acid from Water

    ERIC Educational Resources Information Center

    Loyo-Rosales, Jorge E.; Torrents, Alba; Rosales-Rivera, Georgina C.; Rice, Clifford C.

    2006-01-01

    Several chemical concepts to the extraction of a water pollutant OPC (octylphenoxyacetic acid) is presented. As an introduction to the laboratory experiment, a discussion on endocrine disrupters is conducted to familiarize the student with the background of the experiment and to explain the need for the extraction and quantitation of the OPC which…

  6. Validated Method for the Characterization and Quantification of Extractable and Nonextractable Ellagitannins after Acid Hydrolysis in Pomegranate Fruits, Juices, and Extracts.

    PubMed

    García-Villalba, Rocío; Espín, Juan Carlos; Aaby, Kjersti; Alasalvar, Cesarettin; Heinonen, Marina; Jacobs, Griet; Voorspoels, Stefan; Koivumäki, Tuuli; Kroon, Paul A; Pelvan, Ebru; Saha, Shikha; Tomás-Barberán, Francisco A

    2015-07-29

    Pomegranates are one of the main highly valuable sources of ellagitannins. Despite the potential health benefits of these compounds, reliable data on their content in pomegranates and derived extracts and food products is lacking, as it is usually underestimated due to their complexity, diversity, and lack of commercially available standards. This study describes a new method for the analysis of the extractable and nonextractable ellagitannins based on the quantification of the acid hydrolysis products that include ellagic acid, gallic acid, sanguisorbic acid dilactone, valoneic acid dilactone, and gallagic acid dilactone in pomegranate samples. The study also shows the occurrence of ellagitannin C-glycosides in pomegranates. The method was optimized using a pomegranate peel extract. To quantify nonextractable ellagitannins, freeze-dried pomegranate fruit samples were directly hydrolyzed with 4 M HCl in water at 90 °C for 24 h followed by extraction of the pellet with dimethyl sulfoxide/methanol (50:50, v/v). The method was validated and reproducibility was assessed by means of an interlaboratory trial, showing high reproducibility across six laboratories with relative standard deviations below 15%. Their applicability was demonstrated in several pomegranate extracts, different parts of pomegranate fruit (husk, peels, and mesocarp), and commercial juices. A large variability has been found in the ellagitannin content (150-750 mg of hydrolysis products/g) and type (gallagic acid/ellagic acid ratios between 4 and 0.15) of the 11 pomegranate extracts studied.

  7. Solvent-free microwave extraction of essential oil from aromatic herbs: from laboratory to pilot and industrial scale.

    PubMed

    Filly, Aurore; Fernandez, Xavier; Minuti, Matteo; Visinoni, Francesco; Cravotto, Giancarlo; Chemat, Farid

    2014-05-01

    Solvent-free microwave extraction (SFME) has been proposed as a green method for the extraction of essential oil from aromatic herbs that are extensively used in the food industry. This technique is a combination of microwave heating and dry distillation performed at atmospheric pressure without any added solvent or water. The isolation and concentration of volatile compounds is performed in a single stage. In this work, SFME and a conventional technique, hydro-distillation HD (Clevenger apparatus), are used for the extraction of essential oil from rosemary (Rosmarinus officinalis L.) and are compared. This preliminary laboratory study shows that essential oils extracted by SFME in 30min were quantitatively (yield and kinetics profile) and qualitatively (aromatic profile) similar to those obtained using conventional hydro-distillation in 2h. Experiments performed in a 75L pilot microwave reactor prove the feasibility of SFME up scaling and potential industrial applications. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Comparison of marine sampling methods for organic contaminants: Passive samplers, water extractions, and live oyster deployment.

    PubMed

    Raub, Kristin B; Vlahos, Penny; Whitney, Michael

    2015-08-01

    Laboratory and field trials evaluated the efficacy of three methods of detecting aquatic pesticide concentrations. Currently used pesticides: atrazine, metolachlor, and diazinon and legacy pesticide dieldrin were targeted. Pesticides were extracted using solid-phase extraction (SPE) of water samples, titanium plate passive samplers coated in ethylene vinyl acetate (EVA) and eastern oysters (Crassostrea viginica) as biosamplers. A laboratory study assessed the extraction efficiencies and precision of each method. Passive samplers yielded the highest precision of the three methods (RSD: 3-14% EVA plates; 19-60% oysters; and 25-56% water samples). Equilibrium partition coefficients were derived. A significant relationship was found between the concentration in oyster tissue and the ambient aquatic concentration. In the field (Housatonic River, CT (U.S.)) water sampling (n = 5) detected atrazine at 1.61-7.31 μg L(-1), oyster sampling (n = 2×15) detected dieldrin at n.d.-0.096 μg L(-1) SW and the passive samplers (n = 5×3) detected atrazine at 0.97-3.78 μg L(-1) SW and dieldrin at n.d.-0.68 μg L(-1) SW. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Method of purifying neutral organophosphorus extractants

    DOEpatents

    Horwitz, E. Philip; Gatrone, Ralph C.; Chiarizia, Renato

    1988-01-01

    A method for removing acidic contaminants from neutral mono and bifunctional organophosphorous extractants by contacting the extractant with a macroporous cation exchange resin in the H.sup.+ state followed by contact with a macroporous anion exchange resin in the OH.sup.- state, whereupon the resins take up the acidic contaminants from the extractant, purifying the extractant and improving its extraction capability.

  10. Steroid hormones in environmental matrices: extraction method comparison.

    PubMed

    Andaluri, Gangadhar; Suri, Rominder P S; Graham, Kendon

    2017-11-09

    The U.S. Environmental Protection Agency (EPA) has developed methods for the analysis of steroid hormones in water, soil, sediment, and municipal biosolids by HRGC/HRMS (EPA Method 1698). Following the guidelines provided in US-EPA Method 1698, the extraction methods were validated with reagent water and applied to municipal wastewater, surface water, and municipal biosolids using GC/MS/MS for the analysis of nine most commonly detected steroid hormones. This is the first reported comparison of the separatory funnel extraction (SFE), continuous liquid-liquid extraction (CLLE), and Soxhlet extraction methods developed by the U.S. EPA. Furthermore, a solid phase extraction (SPE) method was also developed in-house for the extraction of steroid hormones from aquatic environmental samples. This study provides valuable information regarding the robustness of the different extraction methods. Statistical analysis of the data showed that SPE-based methods provided better recovery efficiencies and lower variability of the steroid hormones followed by SFE. The analytical methods developed in-house for extraction of biosolids showed a wide recovery range; however, the variability was low (≤ 7% RSD). Soxhlet extraction and CLLE are lengthy procedures and have been shown to provide highly variably recovery efficiencies. The results of this study are guidance for better sample preparation strategies in analytical methods for steroid hormone analysis, and SPE adds to the choice in environmental sample analysis.

  11. Rapid method for direct identification of bacteria in urine and blood culture samples by matrix-assisted laser desorption ionization time-of-flight mass spectrometry: intact cell vs. extraction method.

    PubMed

    Ferreira, L; Sánchez-Juanes, F; Muñoz-Bellido, J L; González-Buitrago, J M

    2011-07-01

    Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a fast and reliable technology for the identification of microorganisms with proteomics approaches. Here, we compare an intact cell method and a protein extraction method before application on the MALDI plate for the direct identification of microorganisms in both urine and blood culture samples from clinical microbiology laboratories. The results show that the intact cell method provides excellent results for urine and is a good initial method for blood cultures. The extraction method complements the intact cell method, improving microorganism identification from blood culture. Thus, we consider that MALDI-TOF MS performed directly on urine and blood culture samples, with the protocols that we propose, is a suitable technique for microorganism identification, as compared with the routine methods used in the clinical microbiology laboratory. © 2010 The Authors. Clinical Microbiology and Infection © 2010 European Society of Clinical Microbiology and Infectious Diseases.

  12. Evaluation and comparison of FTA card and CTAB DNA extraction methods for non-agricultural taxa1

    PubMed Central

    Siegel, Chloe S.; Stevenson, Florence O.; Zimmer, Elizabeth A.

    2017-01-01

    Premise of the study: An efficient, effective DNA extraction method is necessary for comprehensive analysis of plant genomes. This study analyzed the quality of DNA obtained using paper FTA cards prepared directly in the field when compared to the more traditional cetyltrimethylammonium bromide (CTAB)–based extraction methods from silica-dried samples. Methods: DNA was extracted using FTA cards according to the manufacturer’s protocol. In parallel, CTAB-based extractions were done using the automated AutoGen DNA isolation system. DNA quality for both methods was determined for 15 non-agricultural species collected in situ, by gel separation, spectrophotometry, fluorometry, and successful amplification and sequencing of nuclear and chloroplast gene markers. Results: The FTA card extraction method yielded less concentrated, but also less fragmented samples than the CTAB-based technique. The card-extracted samples provided DNA that could be successfully amplified and sequenced. The FTA cards are also useful because the collected samples do not require refrigeration, extensive laboratory expertise, or as many hazardous chemicals as extractions using the CTAB-based technique. Discussion: The relative success of the FTA card method in our study suggested that this method could be a valuable tool for studies in plant population genetics and conservation biology that may involve screening of hundreds of individual plants. The FTA cards, like the silica gel samples, do not contain plant material capable of propagation, and therefore do not require permits from the U.S. Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS) for transportation. PMID:28224056

  13. The Effect of Storage and Extraction Methods on Amplification of Plasmodium falciparum DNA from Dried Blood Spots.

    PubMed

    Schwartz, Alanna; Baidjoe, Amrish; Rosenthal, Philip J; Dorsey, Grant; Bousema, Teun; Greenhouse, Bryan

    2015-05-01

    Extraction and amplification of DNA from dried blood spots (DBS) collected in field studies is commonly used for detection of Plasmodium falciparum. However, there have been few systematic efforts to determine the effects of storage and extraction methods on the sensitivity of DNA amplification. We investigated the effects of storage conditions, length of storage, and DNA extraction methods on amplification via three PCR-based assays using field samples and laboratory controls. Samples stored as DBS for 2 or more years at ambient temperature showed a significant loss of sensitivity that increased with time; after 10 years only 10% samples with parasite densities > 1,000 parasites/μL were detectable by nested polymerase chain reaction (PCR). Conversely, DBS and extracted DNA stored at -20°C showed no loss of sensitivity with time. Samples with low parasite densities amplified more successfully with saponin/Chelex compared with spin-column-based extraction, though the latter method performed better on samples with higher parasite densities stored for 2 years at ambient temperature. DNA extracted via both methods was stable after 20 freeze-thaw cycles. Our results suggest that DBS should be stored at -20°C or extracted immediately, especially if anticipating 2 or more years of storage. © The American Society of Tropical Medicine and Hygiene.

  14. The Effect of Storage and Extraction Methods on Amplification of Plasmodium falciparum DNA from Dried Blood Spots

    PubMed Central

    Schwartz, Alanna; Baidjoe, Amrish; Rosenthal, Philip J.; Dorsey, Grant; Bousema, Teun; Greenhouse, Bryan

    2015-01-01

    Extraction and amplification of DNA from dried blood spots (DBS) collected in field studies is commonly used for detection of Plasmodium falciparum. However, there have been few systematic efforts to determine the effects of storage and extraction methods on the sensitivity of DNA amplification. We investigated the effects of storage conditions, length of storage, and DNA extraction methods on amplification via three PCR-based assays using field samples and laboratory controls. Samples stored as DBS for 2 or more years at ambient temperature showed a significant loss of sensitivity that increased with time; after 10 years only 10% samples with parasite densities > 1,000 parasites/μL were detectable by nested polymerase chain reaction (PCR). Conversely, DBS and extracted DNA stored at −20°C showed no loss of sensitivity with time. Samples with low parasite densities amplified more successfully with saponin/Chelex compared with spin-column-based extraction, though the latter method performed better on samples with higher parasite densities stored for 2 years at ambient temperature. DNA extracted via both methods was stable after 20 freeze-thaw cycles. Our results suggest that DBS should be stored at −20°C or extracted immediately, especially if anticipating 2 or more years of storage. PMID:25758652

  15. Extracting metalworking fluid aerosol samples in cassettes by provisional ASTM and NIOSH methods.

    PubMed

    Harper, Martin

    2002-01-01

    Recent provisional methods for the determination of metalworking fluid aerosol in workplace air involve a solvent extraction procedure to separate the nonvolatile fraction of the fluid from insoluble material such as metal turnings and dirt. The procedure calls for preweighing a filter (W1) and assembling it into a cassette and taking a sample. In the laboratory the filter is removed from the cassette, desiccated to remove any collected water or other volatile substances, and weighed again (W2). The filter is then extracted in an organic solvent blend, allowed to dry, and weighed a final time (W3). The total weight collected by the filter is given by (W2-W1), and the weight of (nonvolatile) metalworking fluid collected is given by (W2-W3). The extraction step can take place within a cassette housing if it is relatively inert to the solvent blend used. The extraction of four metalworking fluids (straight oil, soluble oil, synthetic and semisynthetic) within disposable polypropylene cassettes was investigated using the same protocol used to evaluate the original method. For all fluids the extraction efficiency was greater than 95% with a precision better than 5%. The mean blank contribution to the extraction step was 16 micrograms. Blanks were also evaluated after storage, and after transport and storage. A small additional blank contribution could be removed by desiccation. The limits of detection and quantitation of the extraction step were calculated to be 28 and 94 micrograms, respectively.

  16. Method development for the analysis of 1,4-dioxane in drinking water using solid-phase extraction and gas chromatography-mass spectrometry.

    PubMed

    Grimmett, Paul E; Munch, Jean W

    2009-01-01

    1,4-Dioxane has been identified as a probable human carcinogen and an emerging contaminant in drinking water. The United States Environmental Protection Agency's (U.S. EPA) National Exposure Research Laboratory (NERL) has developed a method for the analysis of 1,4-dioxane in drinking water at ng/L concentrations. The method consists of an activated carbon solid-phase extraction of 500-mL or 100-mL water samples using dichloromethane as the elution solvent. The extracts are analyzed by gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode. In the NERL laboratory, recovery of 1,4-dioxane ranged from 94-110% in fortified laboratory reagent water and recoveries of 96-102% were demonstrated for fortified drinking water samples. The relative standard deviations for replicate analyses were less than 6% at concentrations exceeding the minimum reporting level.

  17. Multicenter Comparative Evaluation of Five Commercial Methods for Toxoplasma DNA Extraction from Amniotic Fluid▿

    PubMed Central

    Yera, H.; Filisetti, D.; Bastien, P.; Ancelle, T.; Thulliez, P.; Delhaes, L.

    2009-01-01

    Over the past few years, a number of new nucleic acid extraction methods and extraction platforms using chemistry combined with magnetic or silica particles have been developed, in combination with instruments to facilitate the extraction procedure. The objective of the present study was to investigate the suitability of these automated methods for the isolation of Toxoplasma gondii DNA from amniotic fluid (AF). Therefore, three automated procedures were compared to two commercialized manual extraction methods. The MagNA Pure Compact (Roche), BioRobot EZ1 (Qiagen), and easyMAG (bioMérieux) automated procedures were compared to two manual DNA extraction kits, the QIAamp DNA minikit (Qiagen) and the High Pure PCR template preparation kit (Roche). Evaluation was carried out with two specific Toxoplasma PCRs (targeting the 529-bp repeat element), inhibitor search PCRs, and human beta-globin PCRs. The samples each consisted of 4 ml of AF with or without a calibrated Toxoplasma gondii RH strain suspension (0, 1, 2.5, 5, and 25 tachyzoites/ml). All PCR assays were laboratory-developed real-time PCR assays, using either TaqMan or fluorescent resonance energy transfer probes. A total of 1,178 PCRs were performed, including 978 Toxoplasma PCRs. The automated and manual methods were similar in sensitivity for DNA extraction from T. gondii at the highest concentration (25 Toxoplasma gondii cells/ml). However, our results showed that the DNA extraction procedures led to variable efficacy in isolating low concentrations of tachyzoites in AF samples (<5 Toxoplasma gondii cells/ml), a difference that might have repercussions since low parasite concentrations in AF exist and can lead to congenital toxoplasmosis. PMID:19846633

  18. Extracting natural dyes from wool--an evaluation of extraction methods.

    PubMed

    Manhita, Ana; Ferreira, Teresa; Candeias, António; Dias, Cristina Barrocas

    2011-05-01

    The efficiency of eight different procedures used for the extraction of natural dyes was evaluated using contemporary wool samples dyed with cochineal, madder, woad, weld, brazilwood and logwood. Comparison was made based on the LC-DAD peak areas of the natural dye's main components which had been extracted from the wool samples. Among the tested methods, an extraction procedure with Na(2)EDTA in water/DMF (1:1, v/v) proved to be the most suitable for the extraction of the studied dyes, which presented a wide range of chemical structures. The identification of the natural dyes used in the making of an eighteenth century Arraiolos carpet was possible using the Na(2)EDTA/DMF extraction of the wool embroidery samples and an LC-DAD-MS methodology. The effectiveness of the Na(2)EDTA/DMF extraction method was particularly observed in the extraction of weld dye components. Nine flavone derivatives previously identified in weld extracts could be identified in a single historical sample, confirming the use of this natural dye in the making of Arraiolos carpets. Indigo and brazilwood were also identified in the samples, and despite the fact that these natural dyes were referred in the historical recipes of Arraiolos dyeing, it is the first time that the use of brazilwood is confirmed. Mordant analysis by ICP-MS identified the widespread use of alum in the dyeing process, but in some samples with darker hues, high amounts of iron were found instead.

  19. Determination of catechins and caffeine in camillia sinensis raw materials, extracts, and dietary supplements by HPLC-uv: single-laboratory validation.

    PubMed

    Roman, Mark C

    2013-01-01

    A rapid method has been developed to quantify seven catechins and caffeine in green tea (Camillia sinensis) raw material and powdered extract, and dietary supplements containing green tea extract. The method utilizes RP HPLC with a phenyl-based stationary phase and gradient elution. Detection is by UV absorbance. The total run time, including column re-equilibration, is 13 min. Single-laboratory validation (SLV) has been performed on the method to determine the repeatability, accuracy, selectivity, LOD, LOQ, ruggedness, and linearity for (+)-catechin, (-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin, (-)-gallocatechin gallate, (-)-epigallocatechin gallate, and (+)-gallocatechin, as well as caffeine. Repeatability precision and recovery results met AOAC guidelines for SLV studies for all catechins and caffeine down to a level of approximately 20 mg/g. Finished products containing high concentrations of minerals require the use of EDTA to prevent decomposition of the catechins.

  20. Aseptic laboratory techniques: plating methods.

    PubMed

    Sanders, Erin R

    2012-05-11

    Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: Perform plating procedures without contaminating media. Isolate single bacterial colonies by the streak

  1. Aseptic Laboratory Techniques: Plating Methods

    PubMed Central

    Sanders, Erin R.

    2012-01-01

    Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the

  2. Anti-ulcer and ulcer healing potentials of Musa sapientum peel extract in the laboratory rodents

    PubMed Central

    Onasanwo, Samuel Adetunji; Emikpe, Benjamin Obukowho; Ajah, Austin Azubuike; Elufioye, Taiwo Olayemi

    2013-01-01

    Background: This study investigated the anti-ulcer and ulcer healing potentials of the methanol extract of Musa sapientum peel in the laboratory rats. Materials and Methods: Methanol extract of the peels on Musa sapientum (MEMS) was evaluated for its anti-ulcer using alcohol-induced, aspirin-induced, and pyloric ligation-induced models, and for its ulcer healing employing acetic acid-induced ulcer models in rats. Results: The findings from this experiment showed that MEMS (50, 100 and 200 mg/kg, b.w.) anti-ulcer and ulcer healing activity (P ≤ 0.05) is dose-dependent. Also, MEMS exhibited healing of the ulcer base in all the treated groups when compared with the control group. Conclusion: The outcomes of this experiment revealed that the anti-ulcer effect of MEMS may be due to its anti-secretory and cyto-protective activity. The healing of the ulcer base might not be unconnected with basic fibroblast growth factors responsible for epithelial regeneration. PMID:23900937

  3. Laboratory assessment of indigenous plant extracts for anti-juvenile hormone activity in Culex quinquefasciatus.

    PubMed

    Saxena, R C; Dixit, O P; Sukumaran, P

    1992-07-01

    Of 15 plants tested, five plant extracts showed anti-juvenile hormone-like activity against laboratory colonised late fourth instar larvae and adult female mosquitoes. Petroleum ether extract of Eichhornia crassipes and acetone extracts of Ageratum conyzoides, Cleome icosandra, Tagetes erectes and Tridax procumbens showed growth inhibitory (P less than 0.001) and juvenile hormone mimicing activity to the treated larvae of C. quinquefasciatus.. Larval pupal intermediates, demalanised pupae, defective egg rafts and adult with deformed flight muscles were few noticeable changes. Biting behaviour was observed to be affected only in Ageratum, Cleome and Tridax extracts (P less than 0.001). Loss of fecundity was observed in the treated mosquitoes but no sterilant effects could be seen. Adults, obtained from larvae exposed to the plant extracts produced significantly shorter egg-rafts (P less than 0.005) than in control.

  4. Validation of a PCR-based method for the detection of various rendered materials in feedstuffs using a forensic DNA extraction kit.

    PubMed

    Myers, Michael J; Yancy, Haile F; Araneta, Michael; Armour, Jennifer; Derr, Janice; Hoostelaere, Lawrence A D; Farmer, Doris; Jackson, Falana; Kiessling, William M; Koch, Henry; Lin, Huahua; Liu, Yan; Mowlds, Gabrielle; Pinero, David; Riter, Ken L; Sedwick, John; Shen, Yuelian; Wetherington, June; Younkins, Ronsha

    2006-01-01

    A method trial was initiated to validate the use of a commercial DNA forensic kit to extract DNA from animal feed as part of a PCR-based method. Four different PCR primer pairs (one bovine pair, one porcine pair, one ovine primer pair, and one multispecies pair) were also evaluated. Each laboratory was required to analyze a total of 120 dairy feed samples either not fortified (control, true negative) or fortified with bovine meat and bone meal, porcine meat and bone meal (PMBM), or lamb meal. Feeds were fortified with the animal meals at a concentration of 0.1% (wt/wt). Ten laboratories participated in this trial, and each laboratory was required to evaluate two different primer pairs, i.e., each PCR primer pair was evaluated by five different laboratories. The method was considered to be validated for a given animal source when three or more laboratories achieved at least 97% accuracy (29 correct of 30 samples for 96.7% accuracy, rounded up to 97%) in detecting the fortified samples for that source. Using this criterion, the method was validated for the bovine primer because three laboratories met the criterion, with an average accuracy of 98.9%. The average false-positive rate was 3.0% in these laboratories. A fourth laboratory was 80% accurate in identifying the samples fortified with bovine meat and bone meal. A fifth laboratory was not able to consistently extract the DNA from the feed samples and did not achieve the criterion for accuracy for either the bovine or multispecies PCR primers. For the porcine primers, the method was validated, with four laboratories meeting the criterion for accuracy with an average accuracy of 99.2%. The fifth laboratory had a 93.3% accuracy outcome for the porcine primer. Collectively, these five laboratories had a 1.3% false-positive rate for the porcine primer. No laboratory was able to meet the criterion for accuracy with the ovine primers, most likely because of problems with the synthesis of the primer pair; none of the

  5. Rapid and reliable high-throughput methods of DNA extraction for use in barcoding and molecular systematics of mushrooms.

    PubMed

    Dentinger, Bryn T M; Margaritescu, Simona; Moncalvo, Jean-Marc

    2010-07-01

    We present two methods for DNA extraction from fresh and dried mushrooms that are adaptable to high-throughput sequencing initiatives, such as DNA barcoding. Our results show that these protocols yield ∼85% sequencing success from recently collected materials. Tests with both recent (<2 year) and older (>100 years) specimens reveal that older collections have low success rates and may be an inefficient resource for populating a barcode database. However, our method of extracting DNA from herbarium samples using small amount of tissue is reliable and could be used for important historical specimens. The application of these protocols greatly reduces time, and therefore cost, of generating DNA sequences from mushrooms and other fungi vs. traditional extraction methods. The efficiency of these methods illustrates that standardization and streamlining of sample processing should be shifted from the laboratory to the field. © 2009 Blackwell Publishing Ltd.

  6. Frequency of data extraction errors and methods to increase data extraction quality: a methodological review.

    PubMed

    Mathes, Tim; Klaßen, Pauline; Pieper, Dawid

    2017-11-28

    Our objective was to assess the frequency of data extraction errors and its potential impact on results in systematic reviews. Furthermore, we evaluated the effect of different extraction methods, reviewer characteristics and reviewer training on error rates and results. We performed a systematic review of methodological literature in PubMed, Cochrane methodological registry, and by manual searches (12/2016). Studies were selected by two reviewers independently. Data were extracted in standardized tables by one reviewer and verified by a second. The analysis included six studies; four studies on extraction error frequency, one study comparing different reviewer extraction methods and two studies comparing different reviewer characteristics. We did not find a study on reviewer training. There was a high rate of extraction errors (up to 50%). Errors often had an influence on effect estimates. Different data extraction methods and reviewer characteristics had moderate effect on extraction error rates and effect estimates. The evidence base for established standards of data extraction seems weak despite the high prevalence of extraction errors. More comparative studies are needed to get deeper insights into the influence of different extraction methods.

  7. Object extraction method for image synthesis

    NASA Astrophysics Data System (ADS)

    Inoue, Seiki

    1991-11-01

    The extraction of component objects from images is fundamentally important for image synthesis. In TV program production, one useful method is the Video-Matte technique for specifying the necessary boundary of an object. This, however, involves some manually intricate and tedious processes. A new method proposed in this paper can reduce the needed level of operator skill and simplify object extraction. The object is automatically extracted by just a simple drawing of a thick boundary line. The basic principle involves a thinning of the thick boundary line binary image using the edge intensity of the original image. This method has many practical advantages, including the simplicity of specifying an object, the high accuracy of thinned-out boundary line, its ease of application to moving images, and the lack of any need for adjustment.

  8. A small-scale, portable method for extracting microplastics from marine sediments.

    PubMed

    Coppock, Rachel L; Cole, Matthew; Lindeque, Penelope K; Queirós, Ana M; Galloway, Tamara S

    2017-11-01

    Microplastics (plastic particles, 0.1 μm-5 mm in size) are widespread marine pollutants, accumulating in benthic sediments and shorelines the world over. To gain a clearer understanding of microplastic availability to marine life, and the risks they pose to the health of benthic communities, ecological processes and food security, it is important to obtain accurate measures of microplastic abundance in marine sediments. To date, methods for extracting microplastics from marine sediments have been disadvantaged by complexity, expense, low extraction efficiencies and incompatibility with very fine sediments. Here we present a new, portable method to separate microplastics from sediments of differing types, using the principle of density floatation. The Sediment-Microplastic Isolation (SMI) unit is a custom-built apparatus which consistently extracted microplastics from sediments in a single step, with a mean efficiency of 95.8% (±SE 1.6%; min 70%, max 100%). Zinc chloride, at a density of 1.5 g cm -3 , was deemed an effective and relatively inexpensive floatation media, allowing fine sediment to settle whilst simultaneously enabling floatation of dense polymers. The method was validated by artificially spiking sediment with low and high density microplastics, and its environmental relevance was further tested by extracting plastics present in natural sediment samples from sites ranging in sediment type; fine silt/clay (mean size 10.25 ± SD 3.02 μm) to coarse sand (mean size 149.3 ± SD 49.9 μm). The method presented here is cheap, reproducible and is easily portable, lending itself for use in the laboratory and in the field, eg. on board research vessels. By employing this method, accurate estimates of microplastic type, distribution and abundance in natural sediments can be achieved, with the potential to further our understanding of the availability of microplastics to benthic organisms. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All

  9. Direct immersion single drop micro-extraction method for multi-class pesticides analysis in mango using GC-MS.

    PubMed

    Pano-Farias, Norma S; Ceballos-Magaña, Silvia G; Muñiz-Valencia, Roberto; Jurado, Jose M; Alcázar, Ángela; Aguayo-Villarreal, Ismael A

    2017-12-15

    Due the negative effects of pesticides on environment and human health, more efficient and environmentally friendly methods are needed. In this sense, a simple, fast, free from memory effects and economical direct-immersion single drop micro-extraction (SDME) method and GC-MS for multi-class pesticides determination in mango samples was developed. Sample pre-treatment using ultrasound-assisted solvent extraction and factors affecting the SDME procedure (extractant solvent, drop volume, stirring rate, ionic strength, time, pH and temperature) were optimized using factorial experimental design. This method presented high sensitive (LOD: 0.14-169.20μgkg -1 ), acceptable precision (RSD: 0.7-19.1%), satisfactory recovery (69-119%) and high enrichment factors (20-722). Several obtained LOQs are below the MRLs established by the European Commission; therefore, the method could be applied for pesticides determination in routing analysis and custom laboratories. Moreover, this method has shown to be suitable for determination of some of the studied pesticides in lime, melon, papaya, banana, tomato, and lettuce. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. DNA extraction on bio-chip: history and preeminence over conventional and solid-phase extraction methods.

    PubMed

    Ayoib, Adilah; Hashim, Uda; Gopinath, Subash C B; Md Arshad, M K

    2017-11-01

    This review covers a developmental progression on early to modern taxonomy at cellular level following the advent of electron microscopy and the advancement in deoxyribonucleic acid (DNA) extraction for expatiation of biological classification at DNA level. Here, we discuss the fundamental values of conventional chemical methods of DNA extraction using liquid/liquid extraction (LLE) followed by development of solid-phase extraction (SPE) methods, as well as recent advances in microfluidics device-based system for DNA extraction on-chip. We also discuss the importance of DNA extraction as well as the advantages over conventional chemical methods, and how Lab-on-a-Chip (LOC) system plays a crucial role for the future achievements.

  11. An alternative method for analysis of food taints using stir bar sorptive extraction.

    PubMed

    Ridgway, Kathy; Lalljie, Sam P D; Smith, Roger M

    2010-09-10

    The determination of taints in food products currently can involve the use of several sample extraction techniques, including direct headspace (DHS), steam distillation extraction (SDE) and more recently solid phase microextraction (SPME). Each of these techniques has disadvantages, such as the use of large volumes of solvents (SDE), or limitations in sensitivity (DHS), or have only been applied to date for determination of individual or specific groups of tainting compounds (SPME). The use of stir bar sorptive extraction (SBSE) has been evaluated as a quantitative screening method for unknown tainting compounds in foods. A range of commonly investigated problem compounds, with a range of physical and chemical properties, were examined. The method was optimised to give the best response for the majority of compounds and the performance was evaluated by examining the accuracy, precision, linearity, limits of detection and quantitation and uncertainties for each analyte. For most compounds SBSE gave the lowest limits of detection compared to steam distillation extraction or direct headspace analysis and in general was better than these established techniques. However, for methyl methacrylate and hexanal no response was observed following stir bar extraction under the optimised conditions. The assays were carried out using a single quadrupole GC-MS in scan mode. A comparison of acquisition modes and instrumentation was performed using standards to illustrate the increase in sensitivity possible using more targeted ion monitoring or a more sensitive high resolution mass spectrometer. This comparison illustrated the usefulness of this approach as an alternative to specialised glassware or expensive instrumentation. SBSE in particular offers a 'greener' extraction method by a large reduction in the use of organic solvents and also minimises the potential for contamination from external laboratory sources, which is of particular concern for taint analysis. Copyright

  12. Development of an Efficient Entire-Capsid-Coding-Region Amplification Method for Direct Detection of Poliovirus from Stool Extracts

    PubMed Central

    Kilpatrick, David R.; Nakamura, Tomofumi; Burns, Cara C.; Bukbuk, David; Oderinde, Soji B.; Oberste, M. Steven; Kew, Olen M.; Pallansch, Mark A.; Shimizu, Hiroyuki

    2014-01-01

    Laboratory diagnosis has played a critical role in the Global Polio Eradication Initiative since 1988, by isolating and identifying poliovirus (PV) from stool specimens by using cell culture as a highly sensitive system to detect PV. In the present study, we aimed to develop a molecular method to detect PV directly from stool extracts, with a high efficiency comparable to that of cell culture. We developed a method to efficiently amplify the entire capsid coding region of human enteroviruses (EVs) including PV. cDNAs of the entire capsid coding region (3.9 kb) were obtained from as few as 50 copies of PV genomes. PV was detected from the cDNAs with an improved PV-specific real-time reverse transcription-PCR system and nucleotide sequence analysis of the VP1 coding region. For assay validation, we analyzed 84 stool extracts that were positive for PV in cell culture and detected PV genomes from 100% of the extracts (84/84 samples) with this method in combination with a PV-specific extraction method. PV could be detected in 2/4 stool extract samples that were negative for PV in cell culture. In PV-positive samples, EV species C viruses were also detected with high frequency (27% [23/86 samples]). This method would be useful for direct detection of PV from stool extracts without using cell culture. PMID:25339406

  13. Extraction and identification of cyclobutanones from irradiated cheese employing a rapid direct solvent extraction method.

    PubMed

    Tewfik, Ihab

    2008-01-01

    2-Alkylcyclobutanones (cyclobutanones) are accepted as chemical markers for irradiated foods containing lipid. However, current extraction procedures (Soxhlet-florisil chromatography) for the isolation of these markers involve a long and tedious clean-up regime prior to gas chromatography-mass spectrophotometry identification. This paper outlines an alternative isolation and clean-up method for the extraction of cyclobutanones in irradiated Camembert cheese. The newly developed direct solvent extraction method enables the efficient screening of large numbers of food samples and is not as resource intensive as the BS EN 1785:1997 method. Direct solvent extraction appears to be a simple, robust method and has the added advantage of a considerably shorter extraction time for the analysis of foods containing lipid.

  14. The impact of different DNA extraction kits and laboratories upon the assessment of human gut microbiota composition by 16S rRNA gene sequencing.

    PubMed

    Kennedy, Nicholas A; Walker, Alan W; Berry, Susan H; Duncan, Sylvia H; Farquarson, Freda M; Louis, Petra; Thomson, John M; Satsangi, Jack; Flint, Harry J; Parkhill, Julian; Lees, Charlie W; Hold, Georgina L

    2014-01-01

    Determining bacterial community structure in fecal samples through DNA sequencing is an important facet of intestinal health research. The impact of different commercially available DNA extraction kits upon bacterial community structures has received relatively little attention. The aim of this study was to analyze bacterial communities in volunteer and inflammatory bowel disease (IBD) patient fecal samples extracted using widely used DNA extraction kits in established gastrointestinal research laboratories. Fecal samples from two healthy volunteers (H3 and H4) and two relapsing IBD patients (I1 and I2) were investigated. DNA extraction was undertaken using MoBio Powersoil and MP Biomedicals FastDNA SPIN Kit for Soil DNA extraction kits. PCR amplification for pyrosequencing of bacterial 16S rRNA genes was performed in both laboratories on all samples. Hierarchical clustering of sequencing data was done using the Yue and Clayton similarity coefficient. DNA extracted using the FastDNA kit and the MoBio kit gave median DNA concentrations of 475 (interquartile range 228-561) and 22 (IQR 9-36) ng/µL respectively (p<0.0001). Hierarchical clustering of sequence data by Yue and Clayton coefficient revealed four clusters. Samples from individuals H3 and I2 clustered by patient; however, samples from patient I1 extracted with the MoBio kit clustered with samples from patient H4 rather than the other I1 samples. Linear modelling on relative abundance of common bacterial families revealed significant differences between kits; samples extracted with MoBio Powersoil showed significantly increased Bacteroidaceae, Ruminococcaceae and Porphyromonadaceae, and lower Enterobacteriaceae, Lachnospiraceae, Clostridiaceae, and Erysipelotrichaceae (p<0.05). This study demonstrates significant differences in DNA yield and bacterial DNA composition when comparing DNA extracted from the same fecal sample with different extraction kits. This highlights the importance of ensuring that samples

  15. Inter-laboratory validation of an inexpensive streamlined method to measure inorganic arsenic in rice grain.

    PubMed

    Chaney, Rufus L; Green, Carrie E; Lehotay, Steven J

    2018-05-04

    With the establishment by CODEX of a 200 ng/g limit of inorganic arsenic (iAs) in polished rice grain, more analyses of iAs will be necessary to ensure compliance in regulatory and trade applications, to assess quality control in commercial rice production, and to conduct research involving iAs in rice crops. Although analytical methods using high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (HPLC-ICP-MS) have been demonstrated for full speciation of As, this expensive and time-consuming approach is excessive when regulations are based only on iAs. We report a streamlined sample preparation and analysis of iAs in powdered rice based on heated extraction with 0.28 M HNO 3 followed by hydride generation (HG) under control of acidity and other simple conditions. Analysis of iAs is then conducted using flow-injection HG and inexpensive ICP-atomic emission spectroscopy (AES) or other detection means. A key innovation compared with previous methods was to increase the acidity of the reagent solution with 4 M HCl (prior to reduction of As 5+ to As 3+ ), which minimized interferences from dimethylarsinic acid. An inter-laboratory method validation was conducted among 12 laboratories worldwide in the analysis of six shared blind duplicates and a NIST Standard Reference Material involving different types of rice and iAs levels. Also, four laboratories used the standard HPLC-ICP-MS method to analyze the samples. The results between the methods were not significantly different, and the Horwitz ratio averaged 0.52 for the new method, which meets official method validation criteria. Thus, the simpler, more versatile, and less expensive method may be used by laboratories for several purposes to accurately determine iAs in rice grain. Graphical abstract Comparison of iAs results from new and FDA methods.

  16. Teaching method validation in the clinical laboratory science curriculum.

    PubMed

    Moon, Tara C; Legrys, Vicky A

    2008-01-01

    With the Clinical Laboratory Improvement Amendment's (CLIA) final rule, the ability of the Clinical Laboratory Scientist (CLS) to perform method validation has become increasingly important. Knowledge of the statistical methods and procedures used in method validation is imperative for clinical laboratory scientists. However, incorporating these concepts in a CLS curriculum can be challenging, especially at a time of limited resources. This paper provides an outline of one approach to addressing these topics in lecture courses and integrating them in the student laboratory and the clinical practicum for direct application.

  17. Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory; arsenic speciation in natural-water samples using laboratory and field methods

    USGS Publications Warehouse

    Garbarino, John R.; Bednar, Anthony J.; Burkhardt, Mark R.

    2002-01-01

    Analytical methods for the determination of arsenite [As(III)], arsenate [As(V)], dimethylarsinate (DMA), monomethylarsonate (MMA), and roxarsone in filtered natural-water samples are described. Various analytical methods can be used for the determination, depending on the arsenic species being determined. Arsenic concentration is determined by using inductively coupled plasma-mass spectrometry (ICP-MS) as an arsenic-specific detector for all methods. Laboratory-speciation methods are described that use an ion chromatographic column to separate the arsenic species; the column length, column packing, and mobile phase are dependent on the species of interest. Regardless of the separation technique, the arsenic species are introduced into plasma by eithe rpneumatic nebulization or arsine generation. Analysis times range from 2 to 8 minutes and method detection limits range from 0.1 to 0.6 microgram-arsenic per liter (ug-As/L), 10 to 60 picograms absolute (for a 100-microliter injection), depending on the arsenic species determined and the analytical method used. A field-generation specciation method also is described that uses a strong anion exchange cartridge to separate As(III) from As(V) in the field. As(III) in the eluate and the As(V) in the cartridge extract are determined by direct nebulization ICP-MS. Methylated arsenic species that also are retained on the cartridge will positively bias As(V) results without further laboratory separations. The method detection limit for field speciation is 0.3 ug-As/L. The distribution of arsenic species must be preserved in the field to eliminate changes caused by photochemical oxidation or metal oxyhydroxide precipitation. Preservation techniques, such as refrigeration, the addition of acides, or the additoin of ethylene-diaminetetraacetic acid (EDTA) and the effects of ambient light were tested. Of the preservatives evaluated, EDTA was found to work best with the laboratory- and field-speciation methods for all sample

  18. ON-SITE SOLID PHRASE EXTRACTION AND LABORATORY ...

    EPA Pesticide Factsheets

    Fragrance materials, such as synthetic musks in aqueous samples, are normally analyzed by GC/MS in the selected ion monitoring (SIM) mode to provide maximum sensitivity after liquid-liquid extraction of I -L samples. A I -L sample, however, usually provides too little analyte for full-scan data acquisition. An on-site extraction method for extracting synthetic musks from 60 L of wastewater effluent has been developed. Such a large sample volume permits high-quality, full-scan mass spectra to be obtained for various synthetic musk compounds. Quantification of these compounds was conveniently achieved from the full-scan data directly, without preparing SIM descriptors for each compound to acquire SIM data. The research focused on in the subtasks is the development and application of state-of the-art technologies to meet the needs of the public, Office of Water, and ORD in the area of Water Quality. Located In the subtasks are the various research projects being performed in support of this Task and more in-depth coverage of each project. Briefly, each project's objective is stated below.Subtask 1: To integrate state-of-the-art technologies (polar organic chemical integrative samplers, advanced solid-phase extraction methodologies with liquid chromatography/electrospray/mass spectrometry) and apply them to studying the sources and fate of a select list of PPCPs. Application and improvement of analytical methodologies that can detect non-volatile, polar, water-sol

  19. A modified method for determining tannin-protein precipitation capacity using accelerated solvent extraction (ASE) and microplate gel filtration.

    PubMed

    McArt, Scott H; Spalinger, Donald E; Kennish, John M; Collins, William B

    2006-06-01

    The protein precipitation assay used by Robbins et al., (1987) Ecology 68:98-107 has been shown to predict successfully the reduction in protein availability to some ruminants due to tannins. The procedure, however, is expensive and laborious, which limits its utility, especially for quantitative ecological or nutritional applications where large numbers of assays may be required. We have modified the method to decrease its cost and increase laboratory efficiency by: (1) automating the extraction by using Accelerated Solvent Extraction (ASE); and (2) by scaling and automating the precipitation reaction, chromatography, and spectrometry with microplate gel filtration and an automated UV-VIS microplate spectrometer. ASE extraction is shown to be as effective at extracting tannins as the hot methanol technique. Additionally, the microplate assay is sensitive and precise. We show that the results from the new technique correspond in a nearly 1:1 relationship to the results of the previous technique. Hence, this method could reliably replace the older method with no loss in relevance to herbivore protein digestion. Moreover, the ASE extraction technique should be applicable to other tannin-protein precipitation assays and possibly other phenolic assays.

  20. Comparability of river suspended-sediment sampling and laboratory analysis methods

    USGS Publications Warehouse

    Groten, Joel T.; Johnson, Gregory D.

    2018-03-06

    Accurate measurements of suspended sediment, a leading water-quality impairment in many Minnesota rivers, are important for managing and protecting water resources; however, water-quality standards for suspended sediment in Minnesota are based on grab field sampling and total suspended solids (TSS) laboratory analysis methods that have underrepresented concentrations of suspended sediment in rivers compared to U.S. Geological Survey equal-width-increment or equal-discharge-increment (EWDI) field sampling and suspended sediment concentration (SSC) laboratory analysis methods. Because of this underrepresentation, the U.S. Geological Survey, in collaboration with the Minnesota Pollution Control Agency, collected concurrent grab and EWDI samples at eight sites to compare results obtained using different combinations of field sampling and laboratory analysis methods.Study results determined that grab field sampling and TSS laboratory analysis results were biased substantially low compared to EWDI sampling and SSC laboratory analysis results, respectively. Differences in both field sampling and laboratory analysis methods caused grab and TSS methods to be biased substantially low. The difference in laboratory analysis methods was slightly greater than field sampling methods.Sand-sized particles had a strong effect on the comparability of the field sampling and laboratory analysis methods. These results indicated that grab field sampling and TSS laboratory analysis methods fail to capture most of the sand being transported by the stream. The results indicate there is less of a difference among samples collected with grab field sampling and analyzed for TSS and concentration of fines in SSC. Even though differences are present, the presence of strong correlations between SSC and TSS concentrations provides the opportunity to develop site specific relations to address transport processes not captured by grab field sampling and TSS laboratory analysis methods.

  1. Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory; determination of pesticides in water by C-18 solid-phase extraction and capillary-column gas chromatography/mass spectrometry with selected-ion monitoring

    USGS Publications Warehouse

    Zaugg, Steven D.; Sandstrom, Mark W.; Smith, Steven G.; Fehlberg, Kevin M.

    1995-01-01

    A method for the isolation of 41 pesticides and pesticide metabolites in natural-water samples using C-18 solid-phase extraction and determination by capillary-column gas chromatography/mass spectrometry with selected-ion monitoring is described. Water samples are filtered to remove suspended particulate matter and then are pumped through disposable solid-phase extraction columns containing octadecyl-bonded porous silica to extract the pesticides. The columns are dried using carbon dioxide or nitrogen gas, and adsorbed pesticides are removed from the columns by elution with 3.0 milliliters of hexane-isopropanol (3:1). Extracted pesticides are determined by capillary- column gas chromatography/mass spectrometry with selected-ion monitoring of three characteristic ions. The upper concentration limit is 4 micrograms per liter (g/L) for most pesticides, with the exception of widely used corn herbicides--atrazine, alachlor, cyanazine, and metolachlor--which have upper concentration limits of 20 g/L. Single- operator method detection limits in reagent-water samples range from 0.001 to 0.018 g/L. Average short-term single-operator precision in reagent- water samples is 7 percent at the 0.1- and 1.0-g/L levels and 8 percent at the 0.01-g/L level. Mean recoveries in reagent-water samples are 73 percent at the 0.1- and 1.0-g/L levels and 83 percent at the 0.01-g/L level. The estimated holding time for pesticides after extraction on the solid-phase extraction columns was 7 days. An optional on-site extraction procedure allows for samples to be collected and processed at remote sites where it is difficult to ship samples to the laboratory within the recommended pre-extraction holding time.

  2. Development of an efficient entire-capsid-coding-region amplification method for direct detection of poliovirus from stool extracts.

    PubMed

    Arita, Minetaro; Kilpatrick, David R; Nakamura, Tomofumi; Burns, Cara C; Bukbuk, David; Oderinde, Soji B; Oberste, M Steven; Kew, Olen M; Pallansch, Mark A; Shimizu, Hiroyuki

    2015-01-01

    Laboratory diagnosis has played a critical role in the Global Polio Eradication Initiative since 1988, by isolating and identifying poliovirus (PV) from stool specimens by using cell culture as a highly sensitive system to detect PV. In the present study, we aimed to develop a molecular method to detect PV directly from stool extracts, with a high efficiency comparable to that of cell culture. We developed a method to efficiently amplify the entire capsid coding region of human enteroviruses (EVs) including PV. cDNAs of the entire capsid coding region (3.9 kb) were obtained from as few as 50 copies of PV genomes. PV was detected from the cDNAs with an improved PV-specific real-time reverse transcription-PCR system and nucleotide sequence analysis of the VP1 coding region. For assay validation, we analyzed 84 stool extracts that were positive for PV in cell culture and detected PV genomes from 100% of the extracts (84/84 samples) with this method in combination with a PV-specific extraction method. PV could be detected in 2/4 stool extract samples that were negative for PV in cell culture. In PV-positive samples, EV species C viruses were also detected with high frequency (27% [23/86 samples]). This method would be useful for direct detection of PV from stool extracts without using cell culture. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  3. Comparative analysis of feature extraction methods in satellite imagery

    NASA Astrophysics Data System (ADS)

    Karim, Shahid; Zhang, Ye; Asif, Muhammad Rizwan; Ali, Saad

    2017-10-01

    Feature extraction techniques are extensively being used in satellite imagery and getting impressive attention for remote sensing applications. The state-of-the-art feature extraction methods are appropriate according to the categories and structures of the objects to be detected. Based on distinctive computations of each feature extraction method, different types of images are selected to evaluate the performance of the methods, such as binary robust invariant scalable keypoints (BRISK), scale-invariant feature transform, speeded-up robust features (SURF), features from accelerated segment test (FAST), histogram of oriented gradients, and local binary patterns. Total computational time is calculated to evaluate the speed of each feature extraction method. The extracted features are counted under shadow regions and preprocessed shadow regions to compare the functioning of each method. We have studied the combination of SURF with FAST and BRISK individually and found very promising results with an increased number of features and less computational time. Finally, feature matching is conferred for all methods.

  4. Fast title extraction method for business documents

    NASA Astrophysics Data System (ADS)

    Katsuyama, Yutaka; Naoi, Satoshi

    1997-04-01

    Conventional electronic document filing systems are inconvenient because the user must specify the keywords in each document for later searches. To solve this problem, automatic keyword extraction methods using natural language processing and character recognition have been developed. However, these methods are slow, especially for japanese documents. To develop a practical electronic document filing system, we focused on the extraction of keyword areas from a document by image processing. Our fast title extraction method can automatically extract titles as keywords from business documents. All character strings are evaluated for similarity by rating points associated with title similarity. We classified these points as four items: character sitting size, position of character strings, relative position among character strings, and string attribution. Finally, the character string that has the highest rating is selected as the title area. The character recognition process is carried out on the selected area. It is fast because this process must recognize a small number of patterns in the restricted area only, and not throughout the entire document. The mean performance of this method is an accuracy of about 91 percent and a 1.8 sec. processing time for an examination of 100 Japanese business documents.

  5. [Bacterial identification methods in the microbiology laboratory].

    PubMed

    Bou, Germán; Fernández-Olmos, Ana; García, Celia; Sáez-Nieto, Juan Antonio; Valdezate, Sylvia

    2011-10-01

    In order to identify the agent responsible of the infectious process and understanding the pathogenic/pathological implications, clinical course, and to implement an effective antimicrobial therapy, a mainstay in the practice of clinical microbiology is the allocation of species to a microbial isolation. In daily routine practice microbiology laboratory phenotypic techniques are applied to achieve this goal. However, they have some limitations that are seen more clearly for some kinds of microorganism. Molecular methods can circumvent some of these limitations, although its implementation is not universal. This is due to higher costs and the level of expertise required for thei implementation, so molecular methods are often centralized in reference laboratories and centers. Recently, proteomics-based methods made an important breakthrough in the field of diagnostic microbiology and will undoubtedly have a major impact on the future organization of the microbiology services. This paper is a short review of the most noteworthy aspects of the three bacterial identification methods described above used in microbiology laboratories. Copyright © 2011 Elsevier España, S.L. All rights reserved.

  6. Susceptibility of Tetranychus urticae Koch to an ethanol extract of Cnidoscolus aconitifolius leaves under laboratory conditions.

    PubMed

    Numa, S; Rodríguez, L; Rodríguez, D; Coy-Barrera, E

    2015-01-01

    One of the main pests of commercial rose crops in Colombia is the phytophagous mite Tetranychus urticae Koch. To manage this pest, synthetic chemicals have traditionally been used, some of which are well known to be potentially toxic to the environment and humans. Therefore, alternative strategies for pest management in greenhouse crops have been developed in recent years, including biological control with natural enemies such as parasitoids, predators and entomopathogenic microorganisms as well as chemical control using plant extracts. Such extracts have shown toxicity to insects, which has positioned them as a common alternative in programs of integrated pest management. The objective of this study was to evaluate the effect of an unfractionated ethanolic extract of Cnidoscolus aconitifolius leaves on adult females of T. urticae under laboratory conditions. The extract was chemically characterized by recording its metabolic profile via liquid chromatography coupled to mass spectrometry, along with tentative metabolite identification. The immersion technique and direct application to rose leaves were used to evaluate the effects of seven doses (10-2,000 µg/mL) of the ethanol extract of C. aconitifolius leaves on T. urticae females under laboratory conditions. The mortality and oviposition of individuals were recorded at 24, 48 and 72 h. It was found that the C. aconitifolius leaf extract reduced fertility and increased mortality in a dose-dependent manner. The main metabolites identified included flavonoid- and sesquiterpene-type compounds, in addition to chromone- and xanthone-type compounds as minor constituents with potential acaricidal effects.

  7. Extracting Communities from Complex Networks by the k-Dense Method

    NASA Astrophysics Data System (ADS)

    Saito, Kazumi; Yamada, Takeshi; Kazama, Kazuhiro

    To understand the structural and functional properties of large-scale complex networks, it is crucial to efficiently extract a set of cohesive subnetworks as communities. There have been proposed several such community extraction methods in the literature, including the classical k-core decomposition method and, more recently, the k-clique based community extraction method. The k-core method, although computationally efficient, is often not powerful enough for uncovering a detailed community structure and it produces only coarse-grained and loosely connected communities. The k-clique method, on the other hand, can extract fine-grained and tightly connected communities but requires a substantial amount of computational load for large-scale complex networks. In this paper, we present a new notion of a subnetwork called k-dense, and propose an efficient algorithm for extracting k-dense communities. We applied our method to the three different types of networks assembled from real data, namely, from blog trackbacks, word associations and Wikipedia references, and demonstrated that the k-dense method could extract communities almost as efficiently as the k-core method, while the qualities of the extracted communities are comparable to those obtained by the k-clique method.

  8. Comparison of methods for nutrient measurement in calcareous soils: Ion-exchange resin bag, capsule, membrane, and chemical extractions

    USGS Publications Warehouse

    Sherrod, S.K.; Belnap, J.; Miller, M.E.

    2002-01-01

    Four methods for measuring quantities of 12 plant-available nutrients were compared using three sandy soils in a series of three experiments. Three of the methods use different ion-exchange resin forms—bags, capsules, and membranes—and the fourth was conventional chemical extraction. The first experiment compared nutrient extraction data from a medium of sand saturated with a nutrient solution. The second and third experiments used Nakai and Sheppard series soils from Canyonlands National Park, which are relatively high in soil carbonates. The second experiment compared nutrient extraction data provided by the four methods from soils equilibrated at two temperatures, “warm” and “cold.” The third experiment extracted nutrients from the same soils in a field equilibration. Our results show that the four extraction techniques are not comparable. This conclusion is due to differences among the methods in the net quantities of nutrients extracted from equivalent soil volumes, in the proportional representation of nutrients within similar soils and treatments, in the measurement of nutrients that were added in known quantities, and even in the order of nutrients ranked by net abundance. We attribute the disparities in nutrient measurement among the different resin forms to interacting effects of the inherent differences in resin exchange capacity, differences among nutrients in their resin affinities, and possibly the relatively short equilibration time for laboratory trials. One constraint for measuring carbonate-related nutrients in high-carbonate soils is the conventional ammonium acetate extraction method, which we suspect of dissolving fine CaCO3 particles that are more abundant in Nakai series soils, resulting in erroneously high Ca2+ estimates. For study of plant-available nutrients, it is important to identify the nutrients of foremost interest and understand differences in their resin sorption dynamics to determine the most appropriate extraction

  9. A method suitable for DNA extraction from humus-rich soil.

    PubMed

    Miao, Tianjin; Gao, Song; Jiang, Shengwei; Kan, Guoshi; Liu, Pengju; Wu, Xianming; An, Yingfeng; Yao, Shuo

    2014-11-01

    A rapid and convenient method for extracting DNA from soil is presented. Soil DNA is extracted by direct cell lysis in the presence of EDTA, SDS, phenol, chloroform and isoamyl alcohol (3-methyl-1-butanol) followed by precipitation with 2-propanol. The extracted DNA is purified by modified DNA purification kit and DNA gel extraction kit. With this method, DNA extracted from humus-rich dark brown forest soil was free from humic substances and, therefore, could be used for efficient PCR amplification and restriction digestion. In contrast, DNA sample extracted with the traditional CTAB-based method had lower yield and purity, and no DNA could be extracted from the same soil sample with a commonly-used commercial soil DNA isolation kit. In addition, this method is time-saving and convenient, providing an efficient choice especially for DNA extraction from humus-rich soils.

  10. Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory - Determination of Wastewater Compounds by Polystyrene-Divinylbenzene Solid-Phase Extraction and Capillary-Column Gas Chromatography/Mass Spectrometry

    USGS Publications Warehouse

    Zaugg, Steven D.; Smith, Steven G.; Schroeder, Michael P.; Barber, Larry B.; Burkhardt, Mark R.

    2002-01-01

    A method for the determination of 67 compounds typically found in domestic and industrial wastewater is described. The method was developed in response to increasing concern over the impact of endocrine-disrupting chemicals in wastewater on aquatic organisms. This method also may be useful for evaluating the impact of combined sanitary and storm-sewer overflow on the water quality of urban streams. The method focuses on the determination of compounds that are an indicator of wastewater or that have been chosen on the basis of their endocrine-disrupting potential or toxicity. These compounds include the alkylphenol ethoxylate nonionic surfactants and their degradates, food additives, fragrances, antioxidants, flame retardants, plasticizers, industrial solvents, disinfectants, fecal sterols, polycyclicaromatic hydrocarbons, and high-use domestic pesticides. Water samples are filtered to remove suspended particulate matter and then are extracted by vacuum through disposable solid-phase cartridges that contain polystyrene-divinylbenzene resin. Cartridges are dried with nitrogen gas, and then sorbed compounds are eluted with dichloromethane-diethyl ether (4:1) and determined by capillary-column gas chromatography/mass spectrometry. Recoveries in reagent-water samples fortified at 4 micrograms per liter averaged 74 percent ? 7 percent relative standard deviation for all method compounds. Initial method detection limits for single-component compounds (excluding hormones and sterols) averaged 0.15 microgram per liter. Samples are preserved by filtration, the addition of 60 grams NaCl, and storage at 4 degrees Celsius. The laboratory has established a sample-holding time (prior to sample extraction) of 14 days from the date of sample collection until a statistically accepted method can be used to determine the effectiveness of these sample-preservation procedures.

  11. 7 CFR 51.1179 - Method of juice extraction.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Method of juice extraction. 51.1179 Section 51.1179 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... of Common Sweet Oranges (citrus Sinensis (l) Osbeck) § 51.1179 Method of juice extraction. The juice...

  12. Determination of phosphate in soil extracts in the field: A green chemistry enzymatic method.

    PubMed

    Campbell, Ellen R; Warsko, Kayla; Davidson, Anna-Marie; Bill Campbell, Wilbur H

    2015-01-01

    Measurement of ortho-phosphate in soil extracts usually involves sending dried samples of soil to a laboratory for analysis and waiting several weeks for the results. Phosphate determination methods often involve use of strong acids, heavy metals, and organic dyes. To overcome limitations of this approach, we have developed a phosphate determination method which can be carried out in the field to obtain results on the spot. This new method uses: •Small volumes.•An enzymatic reaction.•Green chemistry. First, the soil sample is extracted with deionized water and filtered. Next, an aliquot of the soil extract (0.5 mL) is transferred to a disposable cuvette, containing 0.5 mL of reaction mixture [200 mM HEPES, pH 7.6, 20 mM MgCl2, with 80 nmol 2-amino-6-mercapto-7-methylpurine ribonucleoside (MESG) and 1 unit of recombinant purine nucleoside phosphorylase (PNP; EC 2.4.2.1)], mixed, and incubated for 10 min at field temperature. Absorbance of the completed reaction is measured at 360 nm in open-source, portable photometer linked by bluetooth to a smartphone. The phosphate and phosphorus content of the soil is determined by comparison of its absorbance at 360 nm to a previously prepared standard phosphate curve, which is stored in the smartphone app.

  13. Determination of phosphate in soil extracts in the field: A green chemistry enzymatic method

    PubMed Central

    Campbell, Ellen R.; Warsko, Kayla; Davidson, Anna-Marie; (Bill) Campbell, Wilbur H.

    2015-01-01

    Measurement of ortho-phosphate in soil extracts usually involves sending dried samples of soil to a laboratory for analysis and waiting several weeks for the results. Phosphate determination methods often involve use of strong acids, heavy metals, and organic dyes. To overcome limitations of this approach, we have developed a phosphate determination method which can be carried out in the field to obtain results on the spot. This new method uses: • Small volumes. • An enzymatic reaction. • Green chemistry. First, the soil sample is extracted with deionized water and filtered. Next, an aliquot of the soil extract (0.5 mL) is transferred to a disposable cuvette, containing 0.5 mL of reaction mixture [200 mM HEPES, pH 7.6, 20 mM MgCl2, with 80 nmol 2-amino-6-mercapto-7-methylpurine ribonucleoside (MESG) and 1 unit of recombinant purine nucleoside phosphorylase (PNP; EC 2.4.2.1)], mixed, and incubated for 10 min at field temperature. Absorbance of the completed reaction is measured at 360 nm in open-source, portable photometer linked by bluetooth to a smartphone. The phosphate and phosphorus content of the soil is determined by comparison of its absorbance at 360 nm to a previously prepared standard phosphate curve, which is stored in the smartphone app. PMID:26150991

  14. Noncontact methods for measuring water-surface elevations and velocities in rivers: Implications for depth and discharge extraction

    USGS Publications Warehouse

    Nelson, Jonathan M.; Kinzel, Paul J.; McDonald, Richard R.; Schmeeckle, Mark

    2016-01-01

    Recently developed optical and videographic methods for measuring water-surface properties in a noninvasive manner hold great promise for extracting river hydraulic and bathymetric information. This paper describes such a technique, concentrating on the method of infrared videog- raphy for measuring surface velocities and both acoustic (laboratory-based) and laser-scanning (field-based) techniques for measuring water-surface elevations. In ideal laboratory situations with simple flows, appropriate spatial and temporal averaging results in accurate water-surface elevations and water-surface velocities. In test cases, this accuracy is sufficient to allow direct inversion of the governing equations of motion to produce estimates of depth and discharge. Unlike other optical techniques for determining local depth that rely on transmissivity of the water column (bathymetric lidar, multi/hyperspectral correlation), this method uses only water-surface information, so even deep and/or turbid flows can be investigated. However, significant errors arise in areas of nonhydrostatic spatial accelerations, such as those associated with flow over bedforms or other relatively steep obstacles. Using laboratory measurements for test cases, the cause of these errors is examined and both a simple semi-empirical method and computational results are presented that can potentially reduce bathymetric inversion errors.

  15. Implementation of 5S Method for Ergonomic Laboratory

    NASA Astrophysics Data System (ADS)

    Dila Sari, Amarria; Ilma Rahmillah, Fety; Prabowo Aji, Bagus

    2017-06-01

    This article discusses 5S implementation in Work System Design and Ergonomic Laboratory, Department of Industrial Engineering, Islamic University of Indonesia. There are some problems related to equipment settings for activity involving students such as files which is accumulated over the previous year practicum, as well as the movement of waste in the form of time due to the placement of goods that do not fit. Therefore, this study aims to apply the 5S method in DSK & E laboratory to facilitate the work processes and reduce waste. The project is performed by laboratory management using 5S methods in response to continuous improvement (Kaizen). Moreover, some strategy and suggestions are promoted to impose 5S system within the laboratory. As a result, the tidiness and cleanliness can be achieved that lead to the great performance of laboratory users. Score assessment before implementing 5S DSKE laboratory is at 64 (2.56) while the score after implementation is 32 (1.28) and shows an improvement of 50%. This has implications for better use in the laboratory area, save time when looking for tools and materials due to its location and good visual control, as well as improving the culture and spirit of ‘5S’ on staff regarding better working environment

  16. The extraction of essential oil from patchouli leaves (Pogostemon cablin Benth) using microwave hydrodistillation and solvent-free microwave extraction methods

    NASA Astrophysics Data System (ADS)

    Putri, D. K. Y.; Kusuma, H. S.; Syahputra, M. E.; Parasandi, D.; Mahfud, M.

    2017-12-01

    Patchouli plant (Pogostemon cablin Benth) is one of the important essential oil-producing plant, contributes more than 50% of total exports of Indonesia’s essential oil. However, the extraction of patchouli oil that has been done in Indonesia is generally still used conventional methods that require enormous amount of energy, high solvent usage, and long time of extraction. Therefore, in this study, patchouli oil extraction was carried out by using microwave hydrodistillation and solvent-free microwave extraction methods. Based on this research, it is known that the extraction of patchouli oil using microwave hydrodistillation method with longer extraction time (240 min) only produced patchouli oil’s yield 1.2 times greater than solvent-free microwave extraction method which require faster extraction time (120 min). Otherwise the analysis of electric consumption and the environmental impact, the solvent-free microwave extraction method showed a smaller amount when compared with microwave hydrodistillation method. It is conclude that the use of solvent-free microwave extraction method for patchouli oil extraction is suitably method as a new green technique.

  17. Syringe-cartridge solid-phase extraction method for patulin in apple juice.

    PubMed

    Eisele, Thomas A; Gibson, Midori Z

    2003-01-01

    A syringe-cartridge solid-phase extraction (SPE) method was developed for determination of patulin in apple juice. A 2.5 mL portion of test sample was passed through a conditioned macroporous SPE cartridge and washed with 2 mL 1% sodium bicarbonate followed by 2 mL 1% acetic acid. Patulin was eluted with 1 mL 10% ethyl acetate in ethyl ether and determined by reversed-phase liquid chromatography using a mobile phase consisting of 81% acetonitrile, 9% water, and 10% 0.05M potassium phosphate buffer, pH 2.4. Recoveries averaged 92% and the relative standard deviation was 8.0% in test samples spiked with 50 ng/mL patulin. The method appears to be applicable for monitoring apple juice samples to meet the U.S. Food and Drug Administration compliance action level of 50 microg/kg in an industrial quality assurance laboratory environment.

  18. Standardization of Laboratory Methods for the PERCH Study

    PubMed Central

    Karron, Ruth A.; Morpeth, Susan C.; Bhat, Niranjan; Levine, Orin S.; Baggett, Henry C.; Brooks, W. Abdullah; Feikin, Daniel R.; Hammitt, Laura L.; Howie, Stephen R. C.; Knoll, Maria Deloria; Kotloff, Karen L.; Madhi, Shabir A.; Scott, J. Anthony G.; Thea, Donald M.; Adrian, Peter V.; Ahmed, Dilruba; Alam, Muntasir; Anderson, Trevor P.; Antonio, Martin; Baillie, Vicky L.; Dione, Michel; Endtz, Hubert P.; Gitahi, Caroline; Karani, Angela; Kwenda, Geoffrey; Maiga, Abdoul Aziz; McClellan, Jessica; Mitchell, Joanne L.; Morailane, Palesa; Mugo, Daisy; Mwaba, John; Mwansa, James; Mwarumba, Salim; Nyongesa, Sammy; Panchalingam, Sandra; Rahman, Mustafizur; Sawatwong, Pongpun; Tamboura, Boubou; Toure, Aliou; Whistler, Toni; O’Brien, Katherine L.; Murdoch, David R.

    2017-01-01

    Abstract The Pneumonia Etiology Research for Child Health study was conducted across 7 diverse research sites and relied on standardized clinical and laboratory methods for the accurate and meaningful interpretation of pneumonia etiology data. Blood, respiratory specimens, and urine were collected from children aged 1–59 months hospitalized with severe or very severe pneumonia and community controls of the same age without severe pneumonia and were tested with an extensive array of laboratory diagnostic tests. A standardized testing algorithm and standard operating procedures were applied across all study sites. Site laboratories received uniform training, equipment, and reagents for core testing methods. Standardization was further assured by routine teleconferences, in-person meetings, site monitoring visits, and internal and external quality assurance testing. Targeted confirmatory testing and testing by specialized assays were done at a central reference laboratory. PMID:28575358

  19. Development of an extraction method for perchlorate in soils.

    PubMed

    Cañas, Jaclyn E; Patel, Rashila; Tian, Kang; Anderson, Todd A

    2006-03-01

    Perchlorate originates as a contaminant in the environment from its use in solid rocket fuels and munitions. The current US EPA methods for perchlorate determination via ion chromatography using conductivity detection do not include recommendations for the extraction of perchlorate from soil. This study evaluated and identified appropriate conditions for the extraction of perchlorate from clay loam, loamy sand, and sandy soils. Based on the results of this evaluation, soils should be extracted in a dry, ground (mortar and pestle) state with Milli-Q water in a 1 ratio 1 soil ratio water ratio and diluted no more than 5-fold before analysis. When sandy soils were extracted in this manner, the calculated method detection limit was 3.5 microg kg(-1). The findings of this study have aided in the establishment of a standardized extraction method for perchlorate in soil.

  20. Actinide extraction methods

    DOEpatents

    Peterman, Dean R [Idaho Falls, ID; Klaehn, John R [Idaho Falls, ID; Harrup, Mason K [Idaho Falls, ID; Tillotson, Richard D [Moore, ID; Law, Jack D [Pocatello, ID

    2010-09-21

    Methods of separating actinides from lanthanides are disclosed. A regio-specific/stereo-specific dithiophosphinic acid having organic moieties is provided in an organic solvent that is then contacted with an acidic medium containing an actinide and a lanthanide. The method can extend to separating actinides from one another. Actinides are extracted as a complex with the dithiophosphinic acid. Separation compositions include an aqueous phase, an organic phase, dithiophosphinic acid, and at least one actinide. The compositions may include additional actinides and/or lanthanides. A method of producing a dithiophosphinic acid comprising at least two organic moieties selected from aromatics and alkyls, each moiety having at least one functional group is also disclosed. A source of sulfur is reacted with a halophosphine. An ammonium salt of the dithiophosphinic acid product is precipitated out of the reaction mixture. The precipitated salt is dissolved in ether. The ether is removed to yield the dithiophosphinic acid.

  1. Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory; determination of pesticides in water by graphitized carbon-based solid-phase extraction and high-performance liquid chromatography/mass spectrometry

    USGS Publications Warehouse

    Furlong, Edward T.; Anderson, Bruce D.; Werner, Stephen L.; Soliven, Paul P.; Coffey, Laura J.; Burkhardt, Mark R.

    2001-01-01

    In 1996, the U.S. Geological Survey National Water Quality Laboratory (NWQL) developed and implemented a graphitized carbon-based solid-phase extraction and high-performance liquid chromatographic analytical method to determine polar pesticide concentrations in surface- and ground-water samples. Subsequently, the NWQL developed a complementary analysis that uses high-performance liquid chromatography/mass spectrometry to detect, identify, and quantify polar pesticides and pesticide metabolites in filtered water at concentrations as low as 10 nanograms per liter. This new method was designed to improve sensitivity and selectivity over the prior method, and to reduce known interferences from natural organic matter.In this new method, pesticides are extracted from filtered water samples by useing a 0.5-gram graphitized carbon-based solid-phase extraction cartridge, eluted from the cartridge, and concentrations determined by using high-performance liquid chromatography with electrospray ionization-mass spectrometry. The upper concentration limit is 1.000 microgram per liter (μg/L) for most compounds. Single-operator method detection limits in organic-free water samples fortified with pesticides at a concentration of 0.025 μg/L ranged from 0.0019 to 0.022 μg/L for all compounds in the method. The grand mean (mean of mean recoveries for individual compounds) recoveries in organic-free water samples ranged from 72 to 89 percent, fortified with pesticides at three concentrations between 0.025 and 0.5 μg/L. Grand mean recoveries in ground- and surface-water samples ranged from 46 to 119 percent, also fortified with pesticides at three concentrations between 0.025 and 0.5 μg/L. Long-term recoveries from reagent water spikes were used to demonstrate that 38 of 65 compounds can be reported without qualification of the quantitative result across the analytical range of the method. The remaining 27 are reported with qualified estimates of concentration because of greater

  2. Are extraction methods in quantitative assays of pharmacopoeia monographs exhaustive? A comparison with pressurized liquid extraction.

    PubMed

    Basalo, Carlos; Mohn, Tobias; Hamburger, Matthias

    2006-10-01

    The extraction methods in selected monographs of the European and the Swiss Pharmacopoeia were compared to pressurized liquid extraction (PLE) with respect to the yield of constituents to be dosed in the quantitative assay for the respective herbal drugs. The study included five drugs, Belladonnae folium, Colae semen, Boldo folium, Tanaceti herba and Agni casti fructus. They were selected to cover different classes of compounds to be analyzed and different extraction methods to be used according to the monographs. Extraction protocols for PLE were optimized by varying the solvents and number of extraction cycles. In PLE, yields > 97 % of extractable analytes were typically achieved with two extraction cycles. For alkaloid-containing drugs, the addition of ammonia prior to extraction significantly increased the yield and reduced the number of extraction cycles required for exhaustive extraction. PLE was in all cases superior to the extraction protocol of the pharmacopoeia monographs (taken as 100 %), with differences ranging from 108 % in case of parthenolide in Tanaceti herba to 343 % in case of alkaloids in Boldo folium.

  3. Developing a laboratory protocol for asphalt binder recovery.

    DOT National Transportation Integrated Search

    2014-10-01

    Asphalt binder extraction and recovery are common laboratory procedures used to provide material for research and quality : assurance testing. The most common methods of recovery performed today include the Abson method and the rotary evaporator : (o...

  4. Intra-laboratory validation of microplate methods for total phenolic content and antioxidant activity on polyphenolic extracts, and comparison with conventional spectrophotometric methods.

    PubMed

    Bobo-García, Gloria; Davidov-Pardo, Gabriel; Arroqui, Cristina; Vírseda, Paloma; Marín-Arroyo, María R; Navarro, Montserrat

    2015-01-01

    Total phenolic content (TPC) and antioxidant activity (AA) assays in microplates save resources and time, therefore they can be useful to overcome the fact that the conventional methods are time-consuming, labour intensive and use large amounts of reagents. An intra-laboratory validation of the Folin-Ciocalteu microplate method to measure TPC and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) microplate method to measure AA was performed and compared with conventional spectrophotometric methods. To compare the TPC methods, the confidence intervals of a linear regression were used. In the range of 10-70 mg L(-1) of gallic acid equivalents (GAE), both methods were equivalent. To compare the AA methodologies, the F-test and t-test were used in a range from 220 to 320 µmol L(-1) of Trolox equivalents. Both methods had homogeneous variances, and the means were not significantively different. The limits of detection and quantification for the TPC microplate method were 0.74 and 2.24 mg L(-1) GAE and for the DPPH 12.07 and 36.58 µmol L(-1) of Trolox equivalents. The relative standard deviation of the repeatability and reproducibility for both microplate methods were ≤ 6.1%. The accuracy ranged from 88% to 100%. The microplate and the conventional methods are equals in a 95% confidence level. © 2014 Society of Chemical Industry.

  5. A Hybrid Method for Pancreas Extraction from CT Image Based on Level Set Methods

    PubMed Central

    Tan, Hanqing; Fujita, Hiroshi

    2013-01-01

    This paper proposes a novel semiautomatic method to extract the pancreas from abdominal CT images. Traditional level set and region growing methods that request locating initial contour near the final boundary of object have problem of leakage to nearby tissues of pancreas region. The proposed method consists of a customized fast-marching level set method which generates an optimal initial pancreas region to solve the problem that the level set method is sensitive to the initial contour location and a modified distance regularized level set method which extracts accurate pancreas. The novelty in our method is the proper selection and combination of level set methods, furthermore an energy-decrement algorithm and an energy-tune algorithm are proposed to reduce the negative impact of bonding force caused by connected tissue whose intensity is similar with pancreas. As a result, our method overcomes the shortages of oversegmentation at weak boundary and can accurately extract pancreas from CT images. The proposed method is compared to other five state-of-the-art medical image segmentation methods based on a CT image dataset which contains abdominal images from 10 patients. The evaluated results demonstrate that our method outperforms other methods by achieving higher accuracy and making less false segmentation in pancreas extraction. PMID:24066016

  6. Extraction methods of Amaranthus sp. grain oil isolation.

    PubMed

    Krulj, Jelena; Brlek, Tea; Pezo, Lato; Brkljača, Jovana; Popović, Sanja; Zeković, Zoran; Bodroža Solarov, Marija

    2016-08-01

    Amaranthus sp. is a fast-growing crop with well-known beneficial nutritional values (rich in protein, fat, dietary fiber, ash, and minerals, especially calcium and sodium, and containing a higher amount of lysine than conventional cereals). Amaranthus sp. is an underexploited plant source of squalene, a compound of high importance in the food, cosmetic and pharmaceutical industries. This paper has examined the effects of the different extraction methods (Soxhlet, supercritical fluid and accelerated solvent extraction) on the oil and squalene yield of three genotypes of Amaranthus sp. grain. The highest yield of the extracted oil (78.1 g kg(-1) ) and squalene (4.7 g kg(-1) ) in grain was obtained by accelerated solvent extraction (ASE) in genotype 16. Post hoc Tukey's HSD test at 95% confidence limit showed significant differences between observed samples. Principal component analysis (PCA) and cluster analysis (CA) were used for assessing the effect of different genotypes and extraction methods on oil and squalene yield, and also the fatty acid composition profile. Using coupled PCA and CA of observed samples, possible directions for improving the quality of product can be realized. The results of this study indicate that it is very important to choose both the right genotype and the right method of extraction for optimal oil and squalene yield. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  7. A Novel Extraction Method Combining Plasma with a Whole-Blood Fraction Shows Excellent Sensitivity and Reproducibility for Patients at High Risk for Invasive Aspergillosis

    PubMed Central

    Springer, Jan; Schloßnagel, Hannes; Heinz, Werner; Doedt, Thomas; Soeller, Rainer; Einsele, Hermann

    2012-01-01

    Diagnosis of invasive aspergillosis (IA) is still a major problem in routine clinical practice. Early diagnosis is essential for a good patient prognosis. PCR is a highly sensitive method for the detection of nucleic acids and could play an important role in improving the diagnosis of fungal infections. Therefore, a novel DNA extraction method, ultraclean production (UCP), was developed allowing purification of both cellular and cell-free circulating fungal DNA. In this prospective study we evaluated the commercially available UCP extraction system and compared it to an in-house system. Sixty-three patients at high risk for IA were screened twice weekly, and DNA extracted by both methods was cross-analyzed, in triplicate, by two different real-time PCR assays. The negative predictive values were high for all methods (94.3 to 100%), qualifying them as screening methods, but the sensitivity and diagnostic odds ratios were higher using the UCP extraction method. Sensitivity ranged from 33.3 to 66.7% using the in-house extracts to 100% using the UCP extraction method. Most of the unclassified patients showed no positive PCR results; however, single-positive PCR replicates were observed in some cases. These can bear clinical relevance but should be interpreted with additional clinical and laboratory data. The PCR assays from the UCP extracts showed greater reproducibility than the in-house method for probable IA patients. The standardized UCP extraction method yielded superior results, with regard to sensitivity and reproducibility, than the in-house method. This was independent of the PCR assay used to detect fungal DNA in the sample extracts. PMID:22593600

  8. METHOD FOR THE SUPERCRITICAL FLUID EXTRACTION OF SOILS/SEDIMENTS

    EPA Science Inventory

    Supercritical fluid extraction has been publicized as an extraction method which has several advantages over conventional methods, and it is expected to result in substantial cost and labor savings. This study was designed to evaluate the feasibility of using supercritical fluid ...

  9. Laboratory theory and methods for sediment analysis

    USGS Publications Warehouse

    Guy, Harold P.

    1969-01-01

    The diverse character of fluvial sediments makes the choice of laboratory analysis somewhat arbitrary and the pressing of sediment samples difficult. This report presents some theories and methods used by the Water Resources Division for analysis of fluvial sediments to determine the concentration of suspended-sediment samples and the particle-size distribution of both suspended-sediment and bed-material samples. Other analyses related to these determinations may include particle shape, mineral content, and specific gravity, the organic matter and dissolved solids of samples, and the specific weight of soils. The merits and techniques of both the evaporation and filtration methods for concentration analysis are discussed. Methods used for particle-size analysis of suspended-sediment samples may include the sieve pipet, the VA tube-pipet, or the BW tube-VA tube depending on the equipment available, the concentration and approximate size of sediment in the sample, and the settling medium used. The choice of method for most bed-material samples is usually limited to procedures suitable for sand or to some type of visual analysis for large sizes. Several tested forms are presented to help insure a well-ordered system in the laboratory to handle the samples, to help determine the kind of analysis required for each, to conduct the required processes, and to assist in the required computations. Use of the manual should further 'standardize' methods of fluvial sediment analysis among the many laboratories and thereby help to achieve uniformity and precision of the data.

  10. A comparative evaluation of six principal IgY antibody extraction methods.

    PubMed

    Ren, Hao; Yang, Wenjing; Thirumalai, Diraviyam; Zhang, Xiaoying; Schade, Rüdiger

    2016-03-01

    Egg yolk has been considered a promising source of antibodies. Our study was designed to compare six principal IgY extraction methods (water dilution, polyethylene glycol [PEG] precipitation, caprylic acid extraction, chloroform extraction, phenol extraction, and carrageenan extraction), and to assess their relative extraction efficiencies and the purity of the resulting antibodies. The results showed that the organic solvents (chloroform or phenol) minimised the lipid ratio in the egg yolk. The water dilution, PEG precipitation and caprylic acid extraction methods resulted in high yields, and antibodies purified with PEG and carrageenan exhibited high purity. Our results indicate that phenol extraction would be more suitable for preparing high concentrations of IgY for non-therapeutic usage, while the water dilution and carrageenan extraction methods would be more appropriate for use in the preparation of IgY for oral administration. 2016 FRAME.

  11. Establishing a novel automated magnetic bead-based method for the extraction of DNA from a variety of forensic samples.

    PubMed

    Witt, Sebastian; Neumann, Jan; Zierdt, Holger; Gébel, Gabriella; Röscheisen, Christiane

    2012-09-01

    Automated systems have been increasingly utilized for DNA extraction by many forensic laboratories to handle growing numbers of forensic casework samples while minimizing the risk of human errors and assuring high reproducibility. The step towards automation however is not easy: The automated extraction method has to be very versatile to reliably prepare high yields of pure genomic DNA from a broad variety of sample types on different carrier materials. To prevent possible cross-contamination of samples or the loss of DNA, the components of the kit have to be designed in a way that allows for the automated handling of the samples with no manual intervention necessary. DNA extraction using paramagnetic particles coated with a DNA-binding surface is predestined for an automated approach. For this study, we tested different DNA extraction kits using DNA-binding paramagnetic particles with regard to DNA yield and handling by a Freedom EVO(®)150 extraction robot (Tecan) equipped with a Te-MagS magnetic separator. Among others, the extraction kits tested were the ChargeSwitch(®)Forensic DNA Purification Kit (Invitrogen), the PrepFiler™Automated Forensic DNA Extraction Kit (Applied Biosystems) and NucleoMag™96 Trace (Macherey-Nagel). After an extensive test phase, we established a novel magnetic bead extraction method based upon the NucleoMag™ extraction kit (Macherey-Nagel). The new method is readily automatable and produces high yields of DNA from different sample types (blood, saliva, sperm, contact stains) on various substrates (filter paper, swabs, cigarette butts) with no evidence of a loss of magnetic beads or sample cross-contamination. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  12. BMAA extraction of cyanobacteria samples: which method to choose?

    PubMed

    Lage, Sandra; Burian, Alfred; Rasmussen, Ulla; Costa, Pedro Reis; Annadotter, Heléne; Godhe, Anna; Rydberg, Sara

    2016-01-01

    β-N-Methylamino-L-alanine (BMAA), a neurotoxin reportedly produced by cyanobacteria, diatoms and dinoflagellates, is proposed to be linked to the development of neurological diseases. BMAA has been found in aquatic and terrestrial ecosystems worldwide, both in its phytoplankton producers and in several invertebrate and vertebrate organisms that bioaccumulate it. LC-MS/MS is the most frequently used analytical technique in BMAA research due to its high selectivity, though consensus is lacking as to the best extraction method to apply. This study accordingly surveys the efficiency of three extraction methods regularly used in BMAA research to extract BMAA from cyanobacteria samples. The results obtained provide insights into possible reasons for the BMAA concentration discrepancies in previous publications. In addition and according to the method validation guidelines for analysing cyanotoxins, the TCA protein precipitation method, followed by AQC derivatization and LC-MS/MS analysis, is now validated for extracting protein-bound (after protein hydrolysis) and free BMAA from cyanobacteria matrix. BMAA biological variability was also tested through the extraction of diatom and cyanobacteria species, revealing a high variance in BMAA levels (0.0080-2.5797 μg g(-1) DW).

  13. Social network extraction based on Web: 1. Related superficial methods

    NASA Astrophysics Data System (ADS)

    Khairuddin Matyuso Nasution, Mahyuddin

    2018-01-01

    Often the nature of something affects methods to resolve the related issues about it. Likewise, methods to extract social networks from the Web, but involve the structured data types differently. This paper reveals several methods of social network extraction from the same sources that is Web: the basic superficial method, the underlying superficial method, the description superficial method, and the related superficial methods. In complexity we derive the inequalities between methods and so are their computations. In this case, we find that different results from the same tools make the difference from the more complex to the simpler: Extraction of social network by involving co-occurrence is more complex than using occurrences.

  14. Comparison of DNA extraction methods for meat analysis.

    PubMed

    Yalçınkaya, Burhanettin; Yumbul, Eylem; Mozioğlu, Erkan; Akgoz, Muslum

    2017-04-15

    Preventing adulteration of meat and meat products with less desirable or objectionable meat species is important not only for economical, religious and health reasons, but also, it is important for fair trade practices, therefore, several methods for identification of meat and meat products have been developed. In the present study, ten different DNA extraction methods, including Tris-EDTA Method, a modified Cetyltrimethylammonium Bromide (CTAB) Method, Alkaline Method, Urea Method, Salt Method, Guanidinium Isothiocyanate (GuSCN) Method, Wizard Method, Qiagen Method, Zymogen Method and Genespin Method were examined to determine their relative effectiveness for extracting DNA from meat samples. The results show that the salt method is easy to perform, inexpensive and environmentally friendly. Additionally, it has the highest yield among all the isolation methods tested. We suggest this method as an alternative method for DNA isolation from meat and meat products. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. A Comparison of DNA Extraction Methods using Petunia hybrida Tissues

    PubMed Central

    Tamari, Farshad; Hinkley, Craig S.; Ramprashad, Naderia

    2013-01-01

    Extraction of DNA from plant tissue is often problematic, as many plants contain high levels of secondary metabolites that can interfere with downstream applications, such as the PCR. Removal of these secondary metabolites usually requires further purification of the DNA using organic solvents or other toxic substances. In this study, we have compared two methods of DNA purification: the cetyltrimethylammonium bromide (CTAB) method that uses the ionic detergent hexadecyltrimethylammonium bromide and chloroform-isoamyl alcohol and the Edwards method that uses the anionic detergent SDS and isopropyl alcohol. Our results show that the Edwards method works better than the CTAB method for extracting DNA from tissues of Petunia hybrida. For six of the eight tissues, the Edwards method yielded more DNA than the CTAB method. In four of the tissues, this difference was statistically significant, and the Edwards method yielded 27–80% more DNA than the CTAB method. Among the different tissues tested, we found that buds, 4 days before anthesis, had the highest DNA concentrations and that buds and reproductive tissue, in general, yielded higher DNA concentrations than other tissues. In addition, DNA extracted using the Edwards method was more consistently PCR-amplified than that of CTAB-extracted DNA. Based on these results, we recommend using the Edwards method to extract DNA from plant tissues and to use buds and reproductive structures for highest DNA yields. PMID:23997658

  16. Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory; in-bottle acid digestion of whole-water samples

    USGS Publications Warehouse

    Hoffman, G.L.; Fishman, M. J.; Garbarino, J.R.

    1996-01-01

    Water samples for trace-metal determinations routinely have been prepared in open laboratories. For example, the U.S. Geological Survey method I-3485-85 (Extraction Procedure, for Water- Suspended Sediment) is performed in a laboratory hood on a laboratory bench without any special precautions to control airborne contamination. This method tends to be contamination prone for several trace metals primarily because the samples are transferred, acidified, digested, and filtered in an open laboratory environment. To reduce trace-metal contamination of digested water samples, procedures were established that rely on minimizing sample-transfer steps and using a class-100 clean bench during sample filtration. This new procedure involves the following steps: 1. The sample is acidified with HCl directly in the original water-sample bottle. 2. The water-sample bottle with the cap secured is heated in a laboratory oven. 3. The digestate is filtered in a class-100 laminar-flow clean bench. The exact conditions used (that is, oven temperature, time of heating, and filtration methods) for this digestion procedure are described. Comparisons between the previous U.S Geological Survey open-beaker method I-3485-85 and the new in-bottle procedure for synthetic and field-collected water samples are given. When the new procedure is used, blank concentrations for most trace metals determined are reduced significantly.

  17. A method of extracting impervious surface based on rule algorithm

    NASA Astrophysics Data System (ADS)

    Peng, Shuangyun; Hong, Liang; Xu, Quanli

    2018-02-01

    The impervious surface has become an important index to evaluate the urban environmental quality and measure the development level of urbanization. At present, the use of remote sensing technology to extract impervious surface has become the main way. In this paper, a method to extract impervious surface based on rule algorithm is proposed. The main ideas of the method is to use the rule-based algorithm to extract impermeable surface based on the characteristics and the difference which is between the impervious surface and the other three types of objects (water, soil and vegetation) in the seven original bands, NDWI and NDVI. The steps can be divided into three steps: 1) Firstly, the vegetation is extracted according to the principle that the vegetation is higher in the near-infrared band than the other bands; 2) Then, the water is extracted according to the characteristic of the water with the highest NDWI and the lowest NDVI; 3) Finally, the impermeable surface is extracted based on the fact that the impervious surface has a higher NDWI value and the lowest NDVI value than the soil.In order to test the accuracy of the rule algorithm, this paper uses the linear spectral mixed decomposition algorithm, the CART algorithm, the NDII index algorithm for extracting the impervious surface based on six remote sensing image of the Dianchi Lake Basin from 1999 to 2014. Then, the accuracy of the above three methods is compared with the accuracy of the rule algorithm by using the overall classification accuracy method. It is found that the extraction method based on the rule algorithm is obviously higher than the above three methods.

  18. Molluscicidal activity of Physalis angulata L. extracts and fractions on Biomphalaria tenagophila (d'Orbigny, 1835) under laboratory conditions.

    PubMed

    dos Santos, José Augusto A; Tomassini, Therezinha Coelho B; Xavier, Deise Cristina Drummond; Ribeiro, Ivone Maria; da Silva, Melissa Teixeira G; de Morais Filho, Zenildo Buarque

    2003-04-01

    The main objective of this research is to evaluate the molluscicide activity of Physalis angulata L. Biomphalaria tenagophila specimens under laboratory conditions. Extracts and fractions were supplied by the Laborat rio de Qu mica de Produtos Naturais, Farmanguinhos-Fiocruz. Experiments were performed according to the methodology described by the World Health Organization for molluscicide tests using the concentrations from 0.1 to 500 mg/l of the extracts, fractions and of a pool of physalins modified steroids present in this species. The results show that ethyl acetate and acetone extracts from the whole plant, the ethanolic extracts of the roots and the physalins pool from stems and leaves were active. Only the whole plant extracts were available in sufficient quantity for the determination of LD50 and LD90 values.

  19. Summary of water body extraction methods based on ZY-3 satellite

    NASA Astrophysics Data System (ADS)

    Zhu, Yu; Sun, Li Jian; Zhang, Chuan Yin

    2017-12-01

    Extracting from remote sensing images is one of the main means of water information extraction. Affected by spectral characteristics, many methods can be not applied to the satellite image of ZY-3. To solve this problem, we summarize the extraction methods for ZY-3 and analyze the extraction results of existing methods. According to the characteristics of extraction results, the method of WI& single band threshold and the method of texture filtering based on probability statistics are explored. In addition, the advantages and disadvantages of all methods are compared, which provides some reference for the research of water extraction from images. The obtained conclusions are as follows. 1) NIR has higher water sensitivity, consequently when the surface reflectance in the study area is less similar to water, using single band threshold method or multi band operation can obtain the ideal effect. 2) Compared with the water index and HIS optimal index method, object extraction method based on rules, which takes into account not only the spectral information of the water, but also space and texture feature constraints, can obtain better extraction effect, yet the image segmentation process is time consuming and the definition of the rules requires a certain knowledge. 3) The combination of the spectral relationship and water index can eliminate the interference of the shadow to a certain extent. When there is less small water or small water is not considered in further study, texture filtering based on probability statistics can effectively reduce the noises in result and avoid mixing shadows or paddy field with water in a certain extent.

  20. SIMPLIFIED METHOD FOR EXTRACTING BOUND PESTICIDES FROM AVIAN SERUM

    EPA Science Inventory

    A simple solid-phase extraction (SPE) method was developed to extract organochlorine pesticides (OCs) and persistent organic pollutants (POPs) from avian serum. In this method, a 1-mL serum sample fortified with two levels of OCs or POPs was treated with 8M urea or 4M urea and 4...

  1. Influence of different extraction methods on the yield and linalool content of the extracts of Eugenia uniflora L.

    PubMed

    Galhiane, Mário S; Rissato, Sandra R; Chierice, Gilberto O; Almeida, Marcos V; Silva, Letícia C

    2006-09-15

    This work has been developed using a sylvestral fruit tree, native to the Brazilian forest, the Eugenia uniflora L., one of the Mirtaceae family. The main goal of the analytical study was focused on extraction methods themselves. The method development pointed to the Clevenger extraction as the best yield in relation to SFE and Soxhlet. The SFE method presented a good yield but showed a big amount of components in the final extract, demonstrating low selectivity. The essential oil extracted was analyzed by GC/FID showing a large range of polarity and boiling point compounds, where linalool, a widely used compound, was identified. Furthermore, an analytical solid phase extraction method was used to clean it up and obtain separated classes of compounds that were fractionated and studied by GC/FID and GC/MS.

  2. Development and validation of a magnetic solid-phase extraction with high-performance liquid chromatography method for the simultaneous determination of amphetamine and methadone in urine.

    PubMed

    Taghvimi, Arezou; Hamishehkar, Hamed; Ebrahimi, Mahmoud

    2016-06-01

    The simultaneous determination of amphetamine and methadone was carried out by magnetic graphene oxide nanoparticles, a magnetic solid-phase extraction adsorbent, as a new sample treatment technique. The main factors (the amounts of sample volume, amount of adsorbent, type and amount of extraction organic solvent, time of extraction and desorption, pH, the ionic strength of extraction medium, and agitation rate) influencing the extraction efficiency were investigated and optimized. Under the optimized conditions, good linearity was observed in the range of 100-1500 ng/mL for amphetamine and 100-1000 ng/mL for methadone. The method was evaluated for determination of AM and methadone in positive urine samples, satisfactory results were obtained, therefore magnetic solid-phase extraction can be applied as a novel method for the determination of drugs of abuse in forensic laboratories. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Accelerated solvent extraction (ASE) for purification and extraction of silicone passive samplers used for the monitoring of organic pollutants.

    PubMed

    Brockmeyer, Berit; Kraus, Uta R; Theobald, Norbert

    2015-12-01

    Silicone passive samplers have gained an increasing attention as single-phased, practical and robust samplers for monitoring of organic contaminants in the aquatic environment in recent years. However, analytical challenges arise in routine application during the extraction of analytes as silicone oligomers are co-extracted and interfere severely during chemical analyses (e.g. gas chromatographic techniques). In this study, we present a fast, practical pre-cleaning method for silicone passive samplers applying accelerated solvent extraction (ASE) for the removal of silicone oligomers prior to the water deployment (hexane/dichloromethane, 100 °C, 70 min). ASE was also shown to be a very fast (10 min) and efficient extraction method for non-polar contaminants (non-exposed PRC recoveries 66-101 %) sampled by the silicone membrane. For both applications, temperature, extraction time and the solvent used for ASE have been optimized. Purification of the ASE extract was carried out by silica gel and high-pressure liquid size exclusion chromatography (HPLC-SEC). The silicone oligomer content was checked by total reflection X-ray fluorescence spectroscopy (TXRF) in order to confirm the absence of the silicone oligomers prior to analysis of passive sampler extracts. The established method was applied on real silicone samplers from the North- and Baltic Sea and showed no matrix effects during analysis of organic pollutants. Internal laboratory standard recoveries were in the same range for laboratory, transport and exposed samplers (85-126 %).

  4. Testing the application of Teflon/quartz soil solution samplers for DOM sampling in the Critical Zone: Field and laboratory approaches

    NASA Astrophysics Data System (ADS)

    Dolan, E. M.; Perdrial, J. N.; Vazquez, A.; Hernández, S.; Chorover, J.

    2010-12-01

    Elizabeth Dolan1,2, Julia Perdrial3, Angélica Vázquez-Ortega3, Selene Hernández-Ruiz3, Jon Chorover3 1Deptartment of Soil, Environmental, and Atmospheric Science, University of Missouri. 2Biosphere 2, University of Arizona. 3Deptartment of Soil, Water, and Environmental Science, University of Arizona. Abstract: The behavior of dissolved organic matter (DOM) in soil is important to many biogeochemical processes. Extraction methods to obtain DOM from the unsaturated zone remain a current focus of research as different methods can influence the type and concentration of DOM obtained. Thus, the present comparison study involves three methods for soil solution sampling to assess their impact on DOM quantity and quality: 1) aqueous soil extracts, 2) solution yielded from laboratory installed suction cup samplers and 3) solutions from field installed suction cup samplers. All samples were analyzed for dissolved organic carbon and total nitrogen concentrations. Moreover, DOM quality was analyzed using fluorescence, UV-Vis and FTIR spectroscopies. Results indicate higher DOC values for laboratory extracted DOM: 20 mg/L for aqueous soil extracts and 31 mg/L for lab installed samplers compared to 12 mg/L for field installed samplers. Large variations in C/N ratios were also observed ranging from 1.5 in laboratory extracted DOM to 11 in field samples. Fluorescence excitation-emission matrices of DOM solutions obtained for the laboratory extraction methods showed higher intensities in regions typical for fulvic and humic acid-like materials relative to those extracted in the field. Similarly, the molar absorptivity calculated from DOC concentration normalization of UV-Vis absorbance of the laboratory-derived solutions was significantly higher as well, indicating greater aromaticity. The observed differences can be attributed to soil disturbance associated with obtaining laboratory derived solution samples. Our results indicate that laboratory extraction methods are not

  5. Extraction of Maltol from Fraser Fir: A Comparison of Microwave-Assisted Extraction and Conventional Heating Protocols for the Organic Chemistry Laboratory

    ERIC Educational Resources Information Center

    Koch, Andrew S.; Chimento, Clio A.; Berg, Allison N.; Mughal, Farah D.; Spencer, Jean-Paul; Hovland, Douglas E.; Mbadugha, Bessie; Hovland, Allan K.; Eller, Leah R.

    2015-01-01

    Two methods for the extraction of maltol from Fraser fir needles are performed and compared in this two-week experiment. A traditional benchtop extraction using dichloromethane is compared to a microwave-assisted extraction using aqueous ethanol. Students perform both procedures and weigh the merits of each technique. In doing so, students see a…

  6. Analytical difficulties facing today's regulatory laboratories: issues in method validation.

    PubMed

    MacNeil, James D

    2012-08-01

    The challenges facing analytical laboratories today are not unlike those faced in the past, although both the degree of complexity and the rate of change have increased. Challenges such as development and maintenance of expertise, maintenance and up-dating of equipment, and the introduction of new test methods have always been familiar themes for analytical laboratories, but international guidelines for laboratories involved in the import and export testing of food require management of such changes in a context which includes quality assurance, accreditation, and method validation considerations. Decisions as to when a change in a method requires re-validation of the method or on the design of a validation scheme for a complex multi-residue method require a well-considered strategy, based on a current knowledge of international guidance documents and regulatory requirements, as well the laboratory's quality system requirements. Validation demonstrates that a method is 'fit for purpose', so the requirement for validation should be assessed in terms of the intended use of a method and, in the case of change or modification of a method, whether that change or modification may affect a previously validated performance characteristic. In general, method validation involves method scope, calibration-related parameters, method precision, and recovery. Any method change which may affect method scope or any performance parameters will require re-validation. Some typical situations involving change in methods are discussed and a decision process proposed for selection of appropriate validation measures. © 2012 John Wiley & Sons, Ltd.

  7. Tenax extraction as a simple approach to improve environmental risk assessments.

    PubMed

    Harwood, Amanda D; Nutile, Samuel A; Landrum, Peter F; Lydy, Michael J

    2015-07-01

    It is well documented that using exhaustive chemical extractions is not an effective means of assessing exposure of hydrophobic organic compounds in sediments and that bioavailability-based techniques are an improvement over traditional methods. One technique that has shown special promise as a method for assessing the bioavailability of hydrophobic organic compounds in sediment is the use of Tenax-extractable concentrations. A 6-h or 24-h single-point Tenax-extractable concentration correlates to both bioaccumulation and toxicity. This method has demonstrated effectiveness for several hydrophobic organic compounds in various organisms under both field and laboratory conditions. In addition, a Tenax bioaccumulation model was developed for multiple compounds relating 24-h Tenax-extractable concentrations to oligochaete tissue concentrations exposed in both the laboratory and field. This model has demonstrated predictive capacity for additional compounds and species. Use of Tenax-extractable concentrations to estimate exposure is rapid, simple, straightforward, and relatively inexpensive, as well as accurate. Therefore, this method would be an invaluable tool if implemented in risk assessments. © 2015 SETAC.

  8. A validated UHPLC-tandem mass spectrometry method for quantitative analysis of flavonolignans in milk thistle (Silybum marianum) extracts.

    PubMed

    Graf, Tyler N; Cech, Nadja B; Polyak, Stephen J; Oberlies, Nicholas H

    2016-07-15

    Validated methods are needed for the analysis of natural product secondary metabolites. These methods are particularly important to translate in vitro observations to in vivo studies. Herein, a method is reported for the analysis of the key secondary metabolites, a series of flavonolignans and a flavonoid, from an extract prepared from the seeds of milk thistle [Silybum marianum (L.) Gaertn. (Asteraceae)]. This report represents the first UHPLC MS-MS method validated for quantitative analysis of these compounds. The method takes advantage of the excellent resolution achievable with UHPLC to provide a complete analysis in less than 7min. The method is validated using both UV and MS detectors, making it applicable in laboratories with different types of analytical instrumentation available. Lower limits of quantitation achieved with this method range from 0.0400μM to 0.160μM with UV and from 0.0800μM to 0.160μM with MS. The new method is employed to evaluate variability in constituent composition in various commercial S. marianum extracts, and to show that storage of the milk thistle compounds in DMSO leads to degradation. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Bootstrapping Methods Applied for Simulating Laboratory Works

    ERIC Educational Resources Information Center

    Prodan, Augustin; Campean, Remus

    2005-01-01

    Purpose: The aim of this work is to implement bootstrapping methods into software tools, based on Java. Design/methodology/approach: This paper presents a category of software e-tools aimed at simulating laboratory works and experiments. Findings: Both students and teaching staff use traditional statistical methods to infer the truth from sample…

  10. Optimization of the extraction and purification of the compatible solute ectoine from Halomonas elongate in the laboratory experiment of a commercial production project.

    PubMed

    Chen, Ruifeng; Zhu, Lijun; Lv, Lihuo; Yao, Su; Li, Bin; Qian, Junqing

    2017-06-01

    Optimization of compatible solutes (ectoine) extraction and purification from Halomonas elongata cell fermentation had been investigated in the laboratory tests of a large scale commercial production project. After culturing H. elongata cells in developed medium at 28 °C for 23-30 h, we obtained an average yield and biomass of ectoine for 15.9 g/L and 92.9 (OD 600 ), respectively. Cell lysis was performed with acid treatment at moderate high temperature (60-70 °C). The downstream processing operations were designed to be as follows: filtration, desalination, cation exchange, extraction of crude product and three times of refining. Among which the cation exchange and extraction of crude product acquired a high average recovery rate of 95 and 96%; whereas a great loss rate of 19 and 15% was observed during the filtration and desalination, respectively. Combined with the recovering of ectoine from the mother liquor of the three times refining, the average of overall yield (referring to the amount of ectoine synthesized in cells) and purity of final product obtained were 43% and over 98%, respectively. However, key factors that affected the production efficiency were not yields but the time used in the extraction of crude product, involving the crystallization step from water, which spended 24-72 h according to the production scale. Although regarding to the productivity and simplicity on laboratory scale, the method described here can not compete with other investigations, in this study we acquired higher purity of ectoine and provided downstream processes that are capable of operating on industrial scale.

  11. Overview of EPA Pesticide Laboratories and Methods

    EPA Pesticide Factsheets

    EPA operates two pesticide laboratories that provide a variety of technical services to the Agency, other federal and state agencies, tribal groups and other organizations.The labs develop methods and procedures.

  12. A Method for Extracting Important Segments from Documents Using Support Vector Machines

    NASA Astrophysics Data System (ADS)

    Suzuki, Daisuke; Utsumi, Akira

    In this paper we propose an extraction-based method for automatic summarization. The proposed method consists of two processes: important segment extraction and sentence compaction. The process of important segment extraction classifies each segment in a document as important or not by Support Vector Machines (SVMs). The process of sentence compaction then determines grammatically appropriate portions of a sentence for a summary according to its dependency structure and the classification result by SVMs. To test the performance of our method, we conducted an evaluation experiment using the Text Summarization Challenge (TSC-1) corpus of human-prepared summaries. The result was that our method achieved better performance than a segment-extraction-only method and the Lead method, especially for sentences only a part of which was included in human summaries. Further analysis of the experimental results suggests that a hybrid method that integrates sentence extraction with segment extraction may generate better summaries.

  13. Influence of Extraction Methods on the Yield of Steviol Glycosides and Antioxidants in Stevia rebaudiana Extracts.

    PubMed

    Periche, Angela; Castelló, Maria Luisa; Heredia, Ana; Escriche, Isabel

    2015-06-01

    This study evaluated the application of ultrasound techniques and microwave energy, compared to conventional extraction methods (high temperatures at atmospheric pressure), for the solid-liquid extraction of steviol glycosides (sweeteners) and antioxidants (total phenols, flavonoids and antioxidant capacity) from dehydrated Stevia leaves. Different temperatures (from 50 to 100 °C), times (from 1 to 40 min) and microwave powers (1.98 and 3.30 W/g extract) were used. There was a great difference in the resulting yields according to the treatments applied. Steviol glycosides and antioxidants were negatively correlated; therefore, there is no single treatment suitable for obtaining the highest yield in both groups of compounds simultaneously. The greatest yield of steviol glycosides was obtained with microwave energy (3.30 W/g extract, 2 min), whereas, the conventional method (90 °C, 1 min) was the most suitable for antioxidant extraction. Consequently, the best process depends on the subsequent use (sweetener or antioxidant) of the aqueous extract of Stevia leaves.

  14. Analysis of DDT and its metabolites in soil and water samples obtained in the vicinity of a closed-down factory in Bangladesh using various extraction methods.

    PubMed

    Al Mahmud, M N U; Khalil, Farzana; Rahman, Md Musfiqur; Mamun, M I R; Shoeb, Mohammad; Abd El-Aty, A M; Park, Jong-Hyouk; Shin, Ho-Chul; Nahar, Nilufar; Shim, Jae-Han

    2015-12-01

    This study was conducted to monitor the spread of dichlorodiphenyltrichloroethane (DDT) and its metabolites (dichlorodiphenyldichloroethylene (DDE), dichlorodiphenyldichloroethane (DDD)) in soil and water to regions surrounding a closed DDT factory in Bangladesh. This fulfillment was accomplished using inter-method and inter-laboratory validation studies. DDTs (DDT and its metabolites) from soil samples were extracted using microwave-assisted extraction (MAE), supercritical fluid extraction (SFE), and solvent extraction (SE). Inter-laboratory calibration was assessed by SE, and all methods were validated by intra- and inter-day accuracy (expressed as recovery %) and precision (expressed as relative standard deviation (RSD)) in the same laboratory, at three fortified concentrations (n = 4). DDTs extracted from water samples by liquid-liquid partitioning and all samples were analyzed by gas chromatography (GC)-electron capture detector (ECD) and confirmed by GC/mass spectrometry (GC/MS). Linearities expressed as determination coefficients (R (2)) were ≥0.995 for matrix-matched calibrations. The recovery rate was in the range of 72-120 and 83-110%, with <15% RSD in soil and water, respectively. The limit of quantification (LOQ) was 0.0165 mg kg(-1) in soil and 0.132 μg L(-1) in water. Greater quantities of DDTs were extracted from soil using the MAE and SE techniques than with the SFE method. Higher amounts of DDTs were discovered in the southern (2.2-936 × 10(2) mg kg(-1)) or southwestern (86.3-2067 × 10(2) mg kg(-1)) direction from the factory than in the eastern direction (1.0-48.6 × 10(2) mg kg(-1)). An exception was the soil sample collected 50 ft (15.24 m) east (2904 × 10(2) mg kg(-1)) of the factory. The spread of DDTs in the water bodies (0.59-3.01 μg L(-1)) was approximately equal in all directions. We concluded that DDTs might have been dumped randomly around the warehouse after the closing of the factory.

  15. Determination of Ethyl Carbamate in Chinese Yellow Rice Wine by Diatomaceous Earth Extraction and GC/MS Method.

    PubMed

    Wu, Pinggu; Zhang, Liqun; Shen, Xianghong; Wang, Liyuan; Zou, Yan; Zhang, Jing; Tan, Ying; Tang, Jun; Ma, Bingjie; Pan, Xiaodong; Jiang, Wei

    2015-01-01

    A sensitive and rapid analytical method based on alkaline diatomaceous earth extraction followed by GC/MS was developed for the quantitative determination of the toxic contaminant ethyl carbamate (EC) in yellow rice wines. The optimal extraction conditions were investigated. With the application of diatomaceous earth extraction, the damage of organic acids to the capillary column was greatly reduced. By using d5-EC as an internal standard for quantitative analysis of EC, the linearity of the calibration curves was good between 10 and 1000 ng/mL. The LOD and LOQ were 1.7 and 5.0 μg/kg, respectively. The spiked level of EC was 5.0-300 μg/kg, and the average recovery of the spikes was between 78.4 and 98.2%, with an RSD between 4.3 and 8.3%. Upon validation by five laboratories when spiked with 50, 100, and 300 μg/kg, the average respective recoveries were 102.9, 102.2, and 98.7% with a RSD between 0.7 and 8.1%. The validation results demonstrated that the method is fast, simple, selective, and suitable for the determination of EC in yellow rice wines.

  16. Modified salting-out method: high-yield, high-quality genomic DNA extraction from whole blood using laundry detergent.

    PubMed

    Nasiri, H; Forouzandeh, M; Rasaee, M J; Rahbarizadeh, F

    2005-01-01

    Different approaches have been used to extract DNA from whole blood. In most of these methods enzymes (such as proteinase K and RNAse A) or toxic organic solvents (such as phenol or guanidine isothiocyanate) are used. Since these enzymes are expensive, and most of the materials that are used routinely are toxic, it is desirable to apply an efficient DNA extraction procedure that does not require the use of such materials. In this study, genomic DNA was extracted by the salting-out method, but instead of using an analytical-grade enzyme and chemical detergents, as normally used for DNA isolation, a common laundry powder was used. Different concentrations of the powder were tested, and proteins were precipitated by NaCl-saturated distilled water. Finally, DNA precipitation was performed with the use of 96% ethanol. From the results, we conclude that the optimum concentration of laundry powder for the highest yield and purity of isolated DNA is 30 mg/mL. The procedure was optimized, and a final protocol is suggested. Following the same protocol, DNA was extracted from 100 blood samples, and their amounts were found to be >50 microg/mL of whole blood. The integrity of the DNA fragments was confirmed by agarose gel electrophoresis. Furthermore, the extracted DNA was used as a template for PCR reaction. The results obtained from PCR showed that the final solutions of extracted DNA did not contain any inhibitory material for the enzyme used in the PCR reaction, and indicated that the isolated DNA was of good quality. These results show that this method is simple, fast, safe, and cost-effective, and can be used in medical laboratories and research centers. Copyright 2005 Wiley-Liss, Inc.

  17. A novel star extraction method based on modified water flow model

    NASA Astrophysics Data System (ADS)

    Zhang, Hao; Niu, Yanxiong; Lu, Jiazhen; Ouyang, Zibiao; Yang, Yanqiang

    2017-11-01

    Star extraction is the essential procedure for attitude measurement of star sensor. The great challenge for star extraction is to segment star area exactly from various noise and background. In this paper, a novel star extraction method based on Modified Water Flow Model(MWFM) is proposed. The star image is regarded as a 3D terrain. The morphology is adopted for noise elimination and Tentative Star Area(TSA) selection. Star area can be extracted through adaptive water flowing within TSAs. This method can achieve accurate star extraction with improved efficiency under complex conditions such as loud noise and uneven backgrounds. Several groups of different types of star images are processed using proposed method. Comparisons with existing methods are conducted. Experimental results show that MWFM performs excellently under different imaging conditions. The star extraction rate is better than 95%. The star centroid accuracy is better than 0.075 pixels. The time-consumption is also significantly reduced.

  18. Fitting It All In: Adapting a Green Chemistry Extraction Experiment for Inclusion in an Undergraduate Analytical Laboratory

    ERIC Educational Resources Information Center

    Buckley, Heather L.; Beck, Annelise R.; Mulvihill, Martin J.; Douskey, Michelle C.

    2013-01-01

    Several principles of green chemistry are introduced through this experiment designed for use in the undergraduate analytical chemistry laboratory. An established experiment of liquid CO2 extraction of D-limonene has been adapted to include a quantitative analysis by gas chromatography. This facilitates drop-in incorporation of an exciting…

  19. Optimizing pressurized liquid extraction of microbial lipids using the response surface method.

    PubMed

    Cescut, J; Severac, E; Molina-Jouve, C; Uribelarrea, J-L

    2011-01-21

    Response surface methodology (RSM) was used for the determination of optimum extraction parameters to reach maximum lipid extraction yield with yeast. Total lipids were extracted from oleaginous yeast (Rhodotorula glutinis) using pressurized liquid extraction (PLE). The effects of extraction parameters on lipid extraction yield were studied by employing a second-order central composite design. The optimal condition was obtained as three cycles of 15 min at 100°C with a ratio of 144 g of hydromatrix per 100 g of dry cell weight. Different analysis methods were used to compare the optimized PLE method with two conventional methods (Soxhlet and modification of Bligh and Dyer methods) under efficiency, selectivity and reproducibility criteria thanks to gravimetric analysis, GC with flame ionization detector, High Performance Liquid Chromatography linked to Evaporative Light Scattering Detector (HPLC-ELSD) and thin-layer chromatographic analysis. For each sample, the lipid extraction yield with optimized PLE was higher than those obtained with referenced methods (Soxhlet and Bligh and Dyer methods with, respectively, a recovery of 78% and 85% compared to PLE method). Moreover, the use of PLE led to major advantages such as an analysis time reduction by a factor of 10 and solvent quantity reduction by 70%, compared with traditional extraction methods. Copyright © 2010 Elsevier B.V. All rights reserved.

  20. A comparison of hydroxyl radical and hydrogen peroxide generation in ambient particle extracts and laboratory metal solutions

    NASA Astrophysics Data System (ADS)

    Shen, Huiyun; Anastasio, Cort

    2012-01-01

    Generation of reactive oxygen species (ROS) - including superoxide ( rad O 2-), hydrogen peroxide (HOOH), and hydroxyl radical ( rad OH) - has been suggested as one mechanism underlying the adverse health effects caused by ambient particulate matter (PM). In this study we compare HOOH and rad OH production from fine and coarse PM collected at an urban (Fresno) and rural (Westside) site in the San Joaquin Valley (SJV) of California, as well as from laboratory solutions containing dissolved copper or iron. Samples were extracted in a cell-free, phosphate-buffered saline (PBS) solution containing 50 μM ascorbate (Asc). In our laboratory solutions we find that Cu is a potent source of both HOOH and rad OH, with approximately 90% of the electrons that can be donated from Asc ending up in HOOH and rad OH after 4 h. In contrast, in Fe solutions there is no measurable HOOH and only a modest production of rad OH. Soluble Cu in the SJV PM samples is also a dominant source of HOOH and rad OH. In both laboratory copper solutions and extracts of ambient particles we find much more production of HOOH compared to rad OH: e.g., HOOH generation is approximately 30-60 times faster than rad OH generation. The formation of HOOH and rad OH are positively correlated, with roughly 3% and 8% of HOOH converted to rad OH after 4 and 24 h of extraction, respectively. Although the SJV PM produce much more HOOH than rad OH, since rad OH is a much stronger oxidant it is unclear which species might be more important for oxidant-mediated toxicity from PM inhalation.

  1. A Comparison of Hydroxyl Radical and Hydrogen Peroxide Generation in Ambient Particle Extracts and Laboratory Metal Solutions

    PubMed Central

    Shen, Huiyun; Anastasio, Cort

    2011-01-01

    Generation of reactive oxygen species (ROS) – including superoxide (•O2−), hydrogen peroxide (HOOH), and hydroxyl radical (•OH) – has been suggested as one mechanism underlying the adverse health effects caused by ambient particulate matter (PM). In this study we compare HOOH and •OH production from fine and coarse PM collected at an urban (Fresno) and rural (Westside) site in the San Joaquin Valley (SJV) of California, as well as from laboratory solutions containing dissolved copper or iron. Samples were extracted in a cell-free, phosphate-buffered saline (PBS) solution containing 50 μM ascorbate (Asc). In our laboratory solutions we find that Cu is a potent source of both HOOH and •OH, with approximately 90% of the electrons that can be donated from Asc ending up in HOOH and •OH after 4 h. In contrast, in Fe solutions there is no measurable HOOH and only a modest production of •OH. Soluble Cu in the SJV PM samples is also a dominant source of HOOH and •OH. In both laboratory copper solutions and extracts of ambient particles we find much more production of HOOH compared to •OH: e.g., HOOH generation is approximately 30 – 60 times faster than •OH generation. The formation of HOOH and •OH are positively correlated, with roughly 3 % and 8 % of HOOH converted to •OH after 4 and 24 hr of extraction, respectively. Although the SJV PM produce much more HOOH than •OH, since •OH is a much stronger oxidant it is unclear which species might be more important for oxidant-mediated toxicity from PM inhalation. PMID:22267949

  2. Four Different Methods Comparison for Extraction of Astaxanthin from Green Alga Haematococcus pluvialis

    PubMed Central

    Dong, Shengzhao; Huang, Yi; Zhang, Rui; Wang, Shihui; Liu, Yun

    2014-01-01

    Haematococcus pluvialis is one of the potent organisms for production of astaxanthin. Up to now, no efficient method has been achieved due to its thick cell wall hindering solvent extraction of astaxanthin. In this study, four different methods, hydrochloric acid pretreatment followed by acetone extraction (HCl-ACE), hexane/isopropanol (6 : 4, v/v) mixture solvents extraction (HEX-IPA), methanol extraction followed by acetone extraction (MET-ACE, 2-step extraction), and soy-oil extraction, were intensively evaluated for extraction of astaxanthin from H. pluvialis. Results showed that HCl-ACE method could obtain the highest oil yield (33.3 ± 1.1%) and astaxanthin content (19.8 ± 1.1%). Quantitative NMR analysis provided the fatty acid chain profiles of total lipid extracts. In all cases, oleyl chains were predominant, and high amounts of polyunsaturated fatty acid chains were observed and the major fatty acid components were oleic acid (13–35%), linoleic acid (37–43%), linolenic acid (20–31%), and total saturated acid (17–28%). DPPH radical scavenging activity of extract obtained by HCl-ACE was 73.2 ± 1.0%, which is the highest amongst the four methods. The reducing power of extract obtained by four extraction methods was also examined. It was concluded that the proposed extraction method of HCl-ACE in this work allowed efficient astaxanthin extractability with high antioxidant properties. PMID:24574909

  3. Four different methods comparison for extraction of astaxanthin from green alga Haematococcus pluvialis.

    PubMed

    Dong, Shengzhao; Huang, Yi; Zhang, Rui; Wang, Shihui; Liu, Yun

    2014-01-01

    Haematococcus pluvialis is one of the potent organisms for production of astaxanthin. Up to now, no efficient method has been achieved due to its thick cell wall hindering solvent extraction of astaxanthin. In this study, four different methods, hydrochloric acid pretreatment followed by acetone extraction (HCl-ACE), hexane/isopropanol (6:4, v/v) mixture solvents extraction (HEX-IPA), methanol extraction followed by acetone extraction (MET-ACE, 2-step extraction), and soy-oil extraction, were intensively evaluated for extraction of astaxanthin from H. pluvialis. Results showed that HCl-ACE method could obtain the highest oil yield (33.3±1.1%) and astaxanthin content (19.8±1.1%). Quantitative NMR analysis provided the fatty acid chain profiles of total lipid extracts. In all cases, oleyl chains were predominant, and high amounts of polyunsaturated fatty acid chains were observed and the major fatty acid components were oleic acid (13-35%), linoleic acid (37-43%), linolenic acid (20-31%), and total saturated acid (17-28%). DPPH radical scavenging activity of extract obtained by HCl-ACE was 73.2±1.0%, which is the highest amongst the four methods. The reducing power of extract obtained by four extraction methods was also examined. It was concluded that the proposed extraction method of HCl-ACE in this work allowed efficient astaxanthin extractability with high antioxidant properties.

  4. An observational assessment method for aging laboratory rats

    EPA Science Inventory

    The growth of the aging population highlights the need for laboratory animal models to study the basic biological processes ofaging and susceptibility to toxic chemicals and disease. Methods to evaluate health ofaging animals over time are needed, especially efficient methods for...

  5. Comparison of extraction methods for quantifying vitamin E from animal tissues.

    PubMed

    Xu, Zhimin

    2008-12-01

    Four extraction methods: (1) solvent (SOL), (2) ultrasound assisted solvent (UA), (3) saponification and solvent (SP), and (4) saponification and ultrasound assisted solvent (SP-UA), were used in sample preparation for quantifying vitamin E (tocopherols) in chicken liver and plasma samples. The extraction yields of SOL, UA, SP, and SP-UA methods obtained by adding delta-tocopherol as internal reference were 95%, 104%, 65%, and 62% for liver and 98%, 103%, 97%, and 94% for plasma, respectively. The methods with saponification significantly affected the stabilities of tocopherols in liver samples. The measured values of alpha- and gamma-tocopherols using the solvent only extraction (SOL) method were much lower than that using any of the other extraction methods. This indicated that less of the tocopherols in those samples were in a form that could be extracted directly by solvent. The measured value of alpha-tocopherol in the liver sample using the ultrasound assisted solvent (UA) method was 1.5-2.5 times of that obtained from the saponification and solvent (SP) method. The differences in measured values of tocopherols in the plasma samples by using the two methods were not significant. However, the measured value of the saponification and ultrasound assisted solvent (SP-UA) method was lower than either the saponification and solvent (SP) or the ultrasound assisted solvent (UA) method. Also, the reproducibility of the ultrasound assisted solvent (UA) method was greater than any of the saponification methods. Compared with the traditional saponification method, the ultrasound assisted solvent method could effectively extract tocopherols from sample matrix without any chemical degradation reactions, especially for complex animal tissue such as liver.

  6. Quantitative 31P NMR for Simultaneous Trace Analysis of Organophosphorus Pesticides in Aqueous Media Using the Stir Bar Sorptive Extraction Method

    NASA Astrophysics Data System (ADS)

    Ansari, S.; Talebpour, Z.; Molaabasi, F.; Bijanzadeh, H. R.; Khazaeli, S.

    2016-09-01

    The analysis of pesticides in water samples is of primary concern for quality control laboratories due to the toxicity of these compounds and their associated public health risk. A novel analytical method based on stir bar sorptive extraction (SBSE), followed by 31P quantitative nuclear magnetic resonance (31P QNMR), has been developed for simultaneously monitoring and determining four organophosphorus pesticides (OPPs) in aqueous media. The effects of factors on the extraction efficiency of OPPs were investigated using a Draper-Lin small composite design. An optimal sample volume of 4.2 mL, extraction time of 96 min, extraction temperature of 42°C, and desorption time of 11 min were obtained. The results showed reasonable linearity ranges for all pesticides with correlation coefficients greater than 0.9920. The limit of quantification (LOQ) ranged from 0.1 to 2.60 mg/L, and the recoveries of spiked river water samples were from 82 to 94% with relative standard deviation (RSD) values less than 4%. The results show that this method is simple, selective, rapid, and can be applied to other sample matrices.

  7. Improved Methods for Capture, Extraction, and Quantitative Assay of Environmental DNA from Asian Bigheaded Carp (Hypophthalmichthys spp.)

    PubMed Central

    Turner, Cameron R.; Miller, Derryl J.; Coyne, Kathryn J.; Corush, Joel

    2014-01-01

    Indirect, non-invasive detection of rare aquatic macrofauna using aqueous environmental DNA (eDNA) is a relatively new approach to population and biodiversity monitoring. As such, the sensitivity of monitoring results to different methods of eDNA capture, extraction, and detection is being investigated in many ecosystems and species. One of the first and largest conservation programs with eDNA-based monitoring as a central instrument focuses on Asian bigheaded carp (Hypophthalmichthys spp.), an invasive fish spreading toward the Laurentian Great Lakes. However, the standard eDNA methods of this program have not advanced since their development in 2010. We developed new, quantitative, and more cost-effective methods and tested them against the standard protocols. In laboratory testing, our new quantitative PCR (qPCR) assay for bigheaded carp eDNA was one to two orders of magnitude more sensitive than the existing endpoint PCR assays. When applied to eDNA samples from an experimental pond containing bigheaded carp, the qPCR assay produced a detection probability of 94.8% compared to 4.2% for the endpoint PCR assays. Also, the eDNA capture and extraction method we adapted from aquatic microbiology yielded five times more bigheaded carp eDNA from the experimental pond than the standard method, at a per sample cost over forty times lower. Our new, more sensitive assay provides a quantitative tool for eDNA-based monitoring of bigheaded carp, and the higher-yielding eDNA capture and extraction method we describe can be used for eDNA-based monitoring of any aquatic species. PMID:25474207

  8. Improved methods for capture, extraction, and quantitative assay of environmental DNA from Asian bigheaded carp (Hypophthalmichthys spp.).

    PubMed

    Turner, Cameron R; Miller, Derryl J; Coyne, Kathryn J; Corush, Joel

    2014-01-01

    Indirect, non-invasive detection of rare aquatic macrofauna using aqueous environmental DNA (eDNA) is a relatively new approach to population and biodiversity monitoring. As such, the sensitivity of monitoring results to different methods of eDNA capture, extraction, and detection is being investigated in many ecosystems and species. One of the first and largest conservation programs with eDNA-based monitoring as a central instrument focuses on Asian bigheaded carp (Hypophthalmichthys spp.), an invasive fish spreading toward the Laurentian Great Lakes. However, the standard eDNA methods of this program have not advanced since their development in 2010. We developed new, quantitative, and more cost-effective methods and tested them against the standard protocols. In laboratory testing, our new quantitative PCR (qPCR) assay for bigheaded carp eDNA was one to two orders of magnitude more sensitive than the existing endpoint PCR assays. When applied to eDNA samples from an experimental pond containing bigheaded carp, the qPCR assay produced a detection probability of 94.8% compared to 4.2% for the endpoint PCR assays. Also, the eDNA capture and extraction method we adapted from aquatic microbiology yielded five times more bigheaded carp eDNA from the experimental pond than the standard method, at a per sample cost over forty times lower. Our new, more sensitive assay provides a quantitative tool for eDNA-based monitoring of bigheaded carp, and the higher-yielding eDNA capture and extraction method we describe can be used for eDNA-based monitoring of any aquatic species.

  9. DNA-Based Methods in the Immunohematology Reference Laboratory

    PubMed Central

    Denomme, Gregory A

    2010-01-01

    Although hemagglutination serves the immunohematology reference laboratory well, when used alone, it has limited capability to resolve complex problems. This overview discusses how molecular approaches can be used in the immunohematology reference laboratory. In order to apply molecular approaches to immunohematology, knowledge of genes, DNA-based methods, and the molecular bases of blood groups are required. When applied correctly, DNA-based methods can predict blood groups to resolve ABO/Rh discrepancies, identify variant alleles, and screen donors for antigen-negative units. DNA-based testing in immunohematology is a valuable tool used to resolve blood group incompatibilities and to support patients in their transfusion needs. PMID:21257350

  10. Field turbidity method for the determination of lead in acid extracts of dried paint.

    PubMed

    Studabaker, William B; McCombs, Michelle; Sorrell, Kristen; Salmons, Cynthia; Brown, G Gordon; Binstock, David; Gutknecht, William F; Harper, Sharon L

    2010-07-08

    extraction with measurement using the new turbidimetric method, and the other half was analyzed using microwave extraction and measurement by ICP-AES. The average relative percent difference between the turbidimetric method and the ICP-AES method for the 24 sets measured as milligrams of lead per cm(2) is -0.63 +/- 32.5%; the mean difference is -2.1 +/- 7.0 mg lead per cm(2). Non-parametric and parametric statistical tests on these data showed no difference in the results for the two procedures. At the federal regulated level of 1 mg of lead per cm(2) paint, this turbidimetric method meets the performance requirements for EPA's National Lead Laboratory Accreditation Program (NLLAP) of accuracy within +/-20% and has the potential to meet the performance specifications of EPA's RRP rule.

  11. Laboratory evaluation of polychlorinated biphenyls encapsulation methods

    EPA Science Inventory

    Effectiveness and limitations of the encapsulation method for reducing polychlorinated biphenyls (PCBs) concentrations in indoor air and contaminated surface have been evaluated in the laboratory study. Ten coating materials such as epoxy and polyurethane coatings, latex paint, a...

  12. Calculating the Degradation Rate of Individual Proteins Using Xenopus Extract Systems.

    PubMed

    McDowell, Gary S; Philpott, Anna

    2018-05-16

    The Xenopus extract system has been used extensively as a simple, quick, and robust method for assessing the stability of proteins against proteasomal degradation. In this protocol, methods are provided for assessing the half-life of in vitro translated radiolabeled proteins using Xenopus egg or embryo extracts. © 2019 Cold Spring Harbor Laboratory Press.

  13. DNA extraction methods for detecting genetically modified foods: A comparative study.

    PubMed

    Elsanhoty, Rafaat M; Ramadan, Mohamed Fawzy; Jany, Klaus Dieter

    2011-06-15

    The work presented in this manuscript was achieved to compare six different methods for extracting DNA from raw maize and its derived products. The methods that gave higher yield and quality of DNA were chosen to detect the genetic modification in the samples collected from the Egyptian market. The different methods used were evaluated for extracting DNA from maize kernels (without treatment), maize flour (mechanical treatment), canned maize (sweet corn), frozen maize (sweet corn), maize starch, extruded maize, popcorn, corn flacks, maize snacks, and bread made from corn flour (mechanical and thermal treatments). The quality and quantity of the DNA extracted from the standards, containing known percentages of GMO material and from the different food products were evaluated. For qualitative detection of the GMO varieties in foods, the GMOScreen 35S/NOS test kit was used, to screen the genetic modification in the samples. The positive samples for the 35S promoter and/or the NOS terminator were identified by the standard methods adopted by EU. All of the used methods extracted yielded good DNA quality. However, we noted that the purest DNA extract were obtained using the DNA extraction kit (Roche) and this generally was the best method for extracting DNA from most of the maize-derived foods. We have noted that the yield of DNA extracted from maize-derived foods was generally lower in the processed products. The results indicated that 17 samples were positive for the presence of 35S promoter, while 34% from the samples were positive for the genetically modified maize line Bt-176. Copyright © 2010 Elsevier Ltd. All rights reserved.

  14. Laboratory assessment of novel oral anticoagulants: method suitability and variability between coagulation laboratories.

    PubMed

    Helin, Tuukka A; Pakkanen, Anja; Lassila, Riitta; Joutsi-Korhonen, Lotta

    2013-05-01

    Laboratory tests to assess novel oral anticoagulants (NOACs) are under evaluation. Routine monitoring is unnecessary, but under special circumstances bioactivity assessment becomes crucial. We analyzed the effects of NOACs on coagulation tests and the availability of specific assays at different laboratories. Plasma samples spiked with dabigatran (Dabi; 120 and 300 μg/L) or rivaroxaban (Riva; 60, 146, and 305 μg/L) were sent to 115 and 38 European laboratories, respectively. International normalized ratio (INR) and activated partial thromboplastin time (APTT) were analyzed for all samples; thrombin time (TT) was analyzed specifically for Dabi and calibrated anti-activated factor X (anti-Xa) activity for Riva. We compared the results with patient samples. Results of Dabi samples were reported by 73 laboratories (13 INR and 9 APTT reagents) and Riva samples by 22 laboratories (5 INR and 4 APTT reagents). Both NOACs increased INR values; the increase was modest, albeit larger, for Dabi, with higher CV, especially with Quick (vs Owren) methods. Both NOACs dose-dependently prolonged the APTT. Again, the prolongation and CVs were larger for Dabi. The INR and APTT results varied reagent-dependently (P < 0.005), with less prolongation in patient samples. TT results (Dabi) and calibrated anti-Xa results (Riva) were reported by only 11 and 8 laboratories, respectively. The screening tests INR and APTT are suboptimal in assessing NOACs, having high reagent dependence and low sensitivity and specificity. They may provide information, if laboratories recognize their limitations. The variation will likely increase and the sensitivity differ in clinical samples. Specific assays measure NOACs accurately; however, few laboratories applied them. © 2013 American Association for Clinical Chemistry.

  15. Method and apparatus for continuous flow injection extraction analysis

    DOEpatents

    Hartenstein, Steven D.; Siemer, Darryl D.

    1992-01-01

    A method and apparatus for a continuous flow injection batch extraction aysis system is disclosed employing extraction of a component of a first liquid into a second liquid which is a solvent for a component of the first liquid, and is immiscible with the first liquid, and for separating the first liquid from the second liquid subsequent to extraction of the component of the first liquid.

  16. Identification and Quantitation of Sorbitol-Based Nuclear Clarifying Agents Extracted from Common Laboratory and Consumer Plasticware Made of Polypropylene

    PubMed Central

    McDonald, Jeffrey G.; Cummins, Carolyn L.; Barkley, Robert M.; Thompson, Bonne M.; Lincoln, Holly A.

    2009-01-01

    Reported here is the mass spectral identification of sorbitol-based nuclear clarifying agents (NCAs) and the quantitative description of their extractability from common laboratory and household plasticware made of polypropylene. NCAs are frequently added to polypropylene to improve optical clarity, increase performance properties, and aid in the manufacturing process of this plastic. NCA addition makes polypropylene plasticware more aesthetically pleasing to the user and makes the product competitive with other plastic formulations. We show here that several NCAs are readily extracted with either ethanol or water from plastic labware during typical laboratory procedures. Observed levels ranged from a nanogram to micrograms of NCA. NCAs were also detected in extracts from plastic food storage containers; levels ranged from 1to 10 μg in two of the three brands tested. The electron ionization mass spectra for three sorbitol-based nuclear clarifying agents (1,3:2,4-bis-O-(benzylidene)sorbitol, 1,3:2,4-bis-O-(p-methylbenzylidene)sorbitol, 1,3:2,4-bis-O-(3,4-dimethylbenzylidene)sorbitol) are presented for the native and trimethylsilylderivatized compounds together with the collision-induced dissociation mass spectra; gas and liquid chromatographic data are also reported. These NCAs now join other well-known plasticizers such as phthalate esters and bisphenol A as common laboratory contaminants. While the potential toxicity of NCAs in mammalian systems is unknown, the current data provide scientists and consumers the opportunity to make more informed decisions regarding the use of polypropylene plastics. PMID:18533681

  17. Viability testing of material derived from Mycobacterium tuberculosis prior to removal from a Containment Level-III Laboratory as part of a Laboratory Risk Assessment Program

    PubMed Central

    Blackwood, Kym S; Burdz, Tamara V; Turenne, Christine Y; Sharma, Meenu K; Kabani, Amin M; Wolfe, Joyce N

    2005-01-01

    Background In the field of clinical mycobacteriology, Mycobacterium tuberculosis (MTB) can be a difficult organism to manipulate due to the restrictive environment of a containment level 3 (CL3) laboratory. Tests for rapid diagnostic work involving smears and molecular methods do not require CL3 practices after the organism has been rendered non-viable. While it has been assumed that after organism deactivation these techniques can be performed outside of a CL3, no conclusive study has consistently confirmed that the organisms are noninfectious after the theoretical 'deactivation' steps. Previous studies have shown that initial steps (such as heating /chemical fixation) may not consistently kill MTB organisms. Methods An inclusive viability study (n = 226) was undertaken to determine at which point handling of culture extraction materials does not necessitate a CL3 environment. Four different laboratory protocols tested for viability included: standard DNA extractions for IS6110 fingerprinting, crude DNA preparations for PCR by boiling and mechanical lysis, protein extractions, and smear preparations. For each protocol, laboratory staff planted a proportion of the resulting material to Bactec 12B medium that was observed for growth for 8 weeks. Results Of the 208 isolates initially tested, 21 samples grew within the 8-week period. Sixteen (7.7%) of these yielded positive results for MTB that included samples of: deactivated culture resuspensions exposed to 80°C for 20 minutes, smear preparations and protein extractions. Test procedures were consequently modified and tested again (n = 18), resulting in 0% viability. Conclusions This study demonstrates that it cannot be assumed that conventional practices (i.e. smear preparation) or extraction techniques render the organism non-viable. All methodologies, new and existing, should be examined by individual laboratories to validate the safe removal of material derived from MTB to the outside of a CL3 laboratory. This

  18. Laboratory evaluation of methanolic extract of Atlantia monophylla (Family: Rutaceae) against immature stages of mosquitoes and non-target organisms.

    PubMed

    Sivagnaname, N; Kalyanasundaram, M

    2004-02-01

    Methanolic extracts of the leaves of Atlantia monophylla (Rutaceae) were evaluated for mosquitocidal activity against immature stages of three mosquito species, Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti in the laboratory. Larvae of Cx. quinquefasciatus and pupae of An. stephensi were found more susceptible, with LC50 values of 0.14 mg/l and 0.05 mg/l, respectively. Insect growth regulating activity of this extract was more pronounced against Ae. aegypti, with EI50 value 0.002 mg/l. The extract was found safe to aquatic mosquito predators Gambusia affinis, Poecilia reticulata, and Diplonychus indicus, with the respective LC50 values of 23.4, 21.3, and 5.7 mg/l. The results indicate that the mosquitocidal effects of the extract of this plant were comparable to neem extract and certain synthetic chemical larvicides like fenthion, methoprene, etc.

  19. Test of the combined method for extracting spectroscopic factors in N =50 nuclei

    NASA Astrophysics Data System (ADS)

    Walter, David; Cizewski, J. A.; Baugher, T.; Ratkiewicz, A.; Pain, S. D.; Nunes, F. M.; Ahn, S.; Cerizza, G.; Jones, K. L.; Manning, B.; Thornsberry, C.

    2017-09-01

    The single-particle properties of nuclei near shell closures and r-process waiting points can be observed using single-nucleon transfer reactions with beams of rare isotopes. However, approximations have to be made about the final bound state to extract spectroscopic information. An approach to constrain the bound state potential has been proposed by Mukhamedzhanov and Nunes. At peripheral reaction energies ( 5 MeV/u), the ANC for the nucleus can be extracted, and is combined with the same reaction at higher energies ( 40 MeV/u). These combined measurements can constrain the shape of the bound state potential, and the spectroscopic factor can be reliably extracted. To test this method, the 86Kr(d , p) reaction was performed in inverse kinematics with a 35 MeV/u beam at the National Superconducting Cyclotron Laboratory (NSCL) with the ORRUBA and SIDAR arrays of silicon strip detectors coupled to the S800 spectrometer. Successful results supported the measurement of a radioactive ion beam of 84Se at 45 MeV/u at the NSCL to be measured at the end of 2017. Results from the 86Kr(d , p) measurement will be presented as well as preparations for the upcoming 84Se(d , p) measurement. This work is supported in part by the National Science Foundation and U.S. D.O.E.

  20. [Comparison of two nucleic acid extraction methods for norovirus in oysters].

    PubMed

    Yuan, Qiao; Li, Hui; Deng, Xiaoling; Mo, Yanling; Fang, Ling; Ke, Changwen

    2013-04-01

    To explore a convenient and effective method for norovirus nucleic acid extraction from oysters suitable for long-term viral surveillance. Two methods, namely method A (glycine washing and polyethylene glycol precipitation of the virus followed by silica gel centrifugal column) and method B (protease K digestion followed by application of paramagnetic silicon) were compared for their performance in norovirus nucleic acid extraction from oysters. Real-time RT-PCR was used to detect norovirus in naturally infected oysters and in oysters with induced infection. The two methods yielded comparable positive detection rates for the samples, but the recovery rate of the virus was higher with method B than with method A. Method B is a more convenient and rapid method for norovirus nucleic acid extraction from oysters and suitable for long-term surveillance of norovirus.

  1. Application of micro-solid-phase extraction for the on-site extraction of heterocyclic aromatic amines in seawater.

    PubMed

    Basheer, Chanbasha

    2018-04-01

    An efficient on-site extraction technique to determine carcinogenic heterocyclic aromatic amines in seawater has been reported. A micro-solid-phase extraction device placed inside a portable battery-operated pump was used for the on-site extraction of seawater samples. Before on-site applications, parameters that influence the extraction efficiency (extraction time, type of sorbent materials, suitable desorption solvent, desorption time, and sample volume) were investigated and optimized in the laboratory. The developed method was then used for the on-site sampling of heterocyclic aromatic amines determination in seawater samples close to distillation plant. Once the on-site extraction completed, the small extraction device with the analytes was brought back to the laboratory for analysis using high-performance liquid chromatography with fluorescence detection. Based on the optimized conditions, the calibration curves were linear over the concentration range of 0.05-20 μg/L with correlation coefficients up to 0.996. The limits of detection were 0.004-0.026 μg/L, and the reproducibility values were between 1.3 and 7.5%. To evaluate the extraction efficiency, a comparison was made with conventional solid-phase extraction and it was applied to various fortified real seawater samples. The average relative recoveries obtained from the spiked seawater samples varied in the range 79.9-95.2%. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. A Simple and Efficient Method of Extracting DNA from Aged Bones and Teeth.

    PubMed

    Liu, Qiqi; Liu, Liyan; Zhang, Minli; Zhang, Qingzhen; Wang, Qiong; Ding, Xiaoran; Shao, Liting; Zhou, Zhe; Wang, Shengqi

    2018-05-01

    DNA is often difficult to extract from old bones and teeth due to low levels of DNA and high levels of degradation. This study established a simple yet efficient method for extracting DNA from 20 aged bones and teeth (approximately 60 years old). Based on the concentration and STR typing results, the new method of DNA extraction (OM) developed in this study was compared with the PrepFiler™ BTA Forensic DNA Extraction Kit (BM). The total amount of DNA extracted using the OM method was not significantly different from that extracted using the commercial kit (p > 0.05). However, the number of STR loci detected was significantly higher in the samples processed using the OM method than using the BM method (p < 0.05). This study aimed to establish a DNA extraction method for aged bones and teeth to improve the detection rate of STR typing and reduce costs compared to the BM technique. © 2017 American Academy of Forensic Sciences.

  3. A sensitive method for the determination of Sulfonamides in seawater samples by Solid Phase Extraction and UV-Visible spectrophotometry

    NASA Astrophysics Data System (ADS)

    Errayess, Sophia Ait; Lahcen, Abdellatif Ait; Idrissi, Laila; Marcoaldi, Caterina; Chiavarini, Salvatore; Amine, Aziz

    2017-06-01

    The authors have developed a sensitive spectrophotometric method for determination of sulfonamide derivatives such as sulfanilamide (SAA), sulfadiazine (SDZ), sulfacetamide (SCT) sulfamethoxazole (SMX), sulfamerazine (SMR), sulfadimethoxine (SDX), sulfamethiazole (SMT) and Sulfathiazole (STZ). This method is based on the Bratton-Marshall reaction, which involves the diazotization of sulfonamides with sodium nitrite under acidic conditions, followed by coupling with N-(1-naphtyl) ethylenediamine dihydrochloride (NED) to form a pink colored compound. Therefore, the Bratton-Marshall method was modified by optimizing the reaction conditions, which allows us to determine a low concentration range of sulfonamides compared to the reported methods. The limits of detection and quantification obtained were 0.019-0.05 and 0.06-0.16 μg mL- 1, respectively. In comparison with other reported methods using different coupling agents, the proposed method was found to be the most simple and sensitive for sulfonamides determination. In this paper, the modified method was successfully employed for the determination of sulfonamides in drinking water, seawater and pharmaceutical and veterinary formulations. The purpose of this work is to optimize and develop a simple method for extraction and concentration of sulfonamides present as residues in seawater and their quantification with the recommended spectrophotometric method. Solid phase extraction (SPE) of sulfonamides from seawater samples was evaluated using Oasis HLB cartridges (3 mL, 540 mg). The recovery efficiency was investigated in the sulfonamides concentration range comprised between 0.19 and 126 ng mL- 1. The ease of use of this extraction method makes it very useful for routine laboratory work.

  4. Simultaneous extraction and clean-up of polychlorinated biphenyls and their metabolites from small tissue samples using pressurized liquid extraction

    PubMed Central

    Kania-Korwel, Izabela; Zhao, Hongxia; Norstrom, Karin; Li, Xueshu; Hornbuckle, Keri C.; Lehmler, Hans-Joachim

    2008-01-01

    A pressurized liquid extraction-based method for the simultaneous extraction and in situ clean-up of polychlorinated biphenyls (PCBs), hydroxylated (OH)-PCBs and methylsulfonyl (MeSO2)-PCBs from small (< 0.5 gram) tissue samples was developed and validated. Extraction of a laboratory reference material with hexane:dichloromethane:methanol (48:43:9, v/v) and Florisil as fat retainer allowed an efficient recovery of PCBs (78–112%; RSD: 13–37%), OH-PCBs (46±2%; RSD: 4%) and MeSO2-PCBs (89±21%; RSD: 24%). Comparable results were obtained with an established analysis method for PCBs, OH-PCBs and MeSO2-PCBs. PMID:19019378

  5. Non-structural carbohydrates in woody plants compared among laboratories.

    PubMed

    Quentin, Audrey G; Pinkard, Elizabeth A; Ryan, Michael G; Tissue, David T; Baggett, L Scott; Adams, Henry D; Maillard, Pascale; Marchand, Jacqueline; Landhäusser, Simon M; Lacointe, André; Gibon, Yves; Anderegg, William R L; Asao, Shinichi; Atkin, Owen K; Bonhomme, Marc; Claye, Caroline; Chow, Pak S; Clément-Vidal, Anne; Davies, Noel W; Dickman, L Turin; Dumbur, Rita; Ellsworth, David S; Falk, Kristen; Galiano, Lucía; Grünzweig, José M; Hartmann, Henrik; Hoch, Günter; Hood, Sharon; Jones, Joanna E; Koike, Takayoshi; Kuhlmann, Iris; Lloret, Francisco; Maestro, Melchor; Mansfield, Shawn D; Martínez-Vilalta, Jordi; Maucourt, Mickael; McDowell, Nathan G; Moing, Annick; Muller, Bertrand; Nebauer, Sergio G; Niinemets, Ülo; Palacio, Sara; Piper, Frida; Raveh, Eran; Richter, Andreas; Rolland, Gaëlle; Rosas, Teresa; Saint Joanis, Brigitte; Sala, Anna; Smith, Renee A; Sterck, Frank; Stinziano, Joseph R; Tobias, Mari; Unda, Faride; Watanabe, Makoto; Way, Danielle A; Weerasinghe, Lasantha K; Wild, Birgit; Wiley, Erin; Woodruff, David R

    2015-11-01

    Non-structural carbohydrates (NSC) in plant tissue are frequently quantified to make inferences about plant responses to environmental conditions. Laboratories publishing estimates of NSC of woody plants use many different methods to evaluate NSC. We asked whether NSC estimates in the recent literature could be quantitatively compared among studies. We also asked whether any differences among laboratories were related to the extraction and quantification methods used to determine starch and sugar concentrations. These questions were addressed by sending sub-samples collected from five woody plant tissues, which varied in NSC content and chemical composition, to 29 laboratories. Each laboratory analyzed the samples with their laboratory-specific protocols, based on recent publications, to determine concentrations of soluble sugars, starch and their sum, total NSC. Laboratory estimates differed substantially for all samples. For example, estimates for Eucalyptus globulus leaves (EGL) varied from 23 to 116 (mean = 56) mg g(-1) for soluble sugars, 6-533 (mean = 94) mg g(-1) for starch and 53-649 (mean = 153) mg g(-1) for total NSC. Mixed model analysis of variance showed that much of the variability among laboratories was unrelated to the categories we used for extraction and quantification methods (method category R(2) = 0.05-0.12 for soluble sugars, 0.10-0.33 for starch and 0.01-0.09 for total NSC). For EGL, the difference between the highest and lowest least squares means for categories in the mixed model analysis was 33 mg g(-1) for total NSC, compared with the range of laboratory estimates of 596 mg g(-1). Laboratories were reasonably consistent in their ranks of estimates among tissues for starch (r = 0.41-0.91), but less so for total NSC (r = 0.45-0.84) and soluble sugars (r = 0.11-0.83). Our results show that NSC estimates for woody plant tissues cannot be compared among laboratories. The relative changes in NSC between treatments measured within a laboratory

  6. Text-in-context: a method for extracting findings in mixed-methods mixed research synthesis studies.

    PubMed

    Sandelowski, Margarete; Leeman, Jennifer; Knafl, Kathleen; Crandell, Jamie L

    2013-06-01

    Our purpose in this paper is to propose a new method for extracting findings from research reports included in mixed-methods mixed research synthesis studies. International initiatives in the domains of systematic review and evidence synthesis have been focused on broadening the conceptualization of evidence, increased methodological inclusiveness and the production of evidence syntheses that will be accessible to and usable by a wider range of consumers. Initiatives in the general mixed-methods research field have been focused on developing truly integrative approaches to data analysis and interpretation. The data extraction challenges described here were encountered, and the method proposed for addressing these challenges was developed, in the first year of the ongoing (2011-2016) study: Mixed-Methods Synthesis of Research on Childhood Chronic Conditions and Family. To preserve the text-in-context of findings in research reports, we describe a method whereby findings are transformed into portable statements that anchor results to relevant information about sample, source of information, time, comparative reference point, magnitude and significance and study-specific conceptions of phenomena. The data extraction method featured here was developed specifically to accommodate mixed-methods mixed research synthesis studies conducted in nursing and other health sciences, but reviewers might find it useful in other kinds of research synthesis studies. This data extraction method itself constitutes a type of integration to preserve the methodological context of findings when statements are read individually and in comparison to each other. © 2012 Blackwell Publishing Ltd.

  7. The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue

    NASA Astrophysics Data System (ADS)

    Löfgren, Lars; Forsberg, Gun-Britt; Ståhlman, Marcus

    2016-06-01

    In this study we present a simple and rapid method for tissue lipid extraction. Snap-frozen tissue (15-150 mg) is collected in 2 ml homogenization tubes. 500 μl BUME mixture (butanol:methanol [3:1]) is added and automated homogenization of up to 24 frozen samples at a time in less than 60 seconds is performed, followed by a 5-minute single-phase extraction. After the addition of 500 μl heptane:ethyl acetate (3:1) and 500 μl 1% acetic acid a 5-minute two-phase extraction is performed. Lipids are recovered from the upper phase by automated liquid handling using a standard 96-tip robot. A second two-phase extraction is performed using 500 μl heptane:ethyl acetate (3:1). Validation of the method showed that the extraction recoveries for the investigated lipids, which included sterols, glycerolipids, glycerophospholipids and sphingolipids were similar or better than for the Folch method. We also applied the method for lipid extraction of liver and heart and compared the lipid species profiles with profiles generated after Folch and MTBE extraction. We conclude that the BUME method is superior to the Folch method in terms of simplicity, through-put, automation, solvent consumption, economy, health and environment yet delivering lipid recoveries fully comparable to or better than the Folch method.

  8. Method and apparatus for back-extracting metal chelates

    DOEpatents

    Wai, Chien M.; Smart, Neil G.; Lin, Yuehe

    1998-01-01

    A method of extracting metal and metalloid species from a solid or liquid substrate using a supercritical fluid solvent containing one or more chelating agents followed by back-extracting the metal and metalloid species from the metal and metalloid chelates formed thereby. The back-extraction acidic solution is performed utilizing an acidic solution. Upon sufficient exposure of the metal and metalloid chelates to the acidic solution, the metal and metalloid species are released from the chelates into the acid solution, while the chelating agent remains in the supercritical fluid solvent. The chelating agent is thereby regenerated and the metal and metalloid species recovered.

  9. Comparison of water extraction methods in Tibet based on GF-1 data

    NASA Astrophysics Data System (ADS)

    Jia, Lingjun; Shang, Kun; Liu, Jing; Sun, Zhongqing

    2018-03-01

    In this study, we compared four different water extraction methods with GF-1 data according to different water types in Tibet, including Support Vector Machine (SVM), Principal Component Analysis (PCA), Decision Tree Classifier based on False Normalized Difference Water Index (FNDWI-DTC), and PCA-SVM. The results show that all of the four methods can extract large area water body, but only SVM and PCA-SVM can obtain satisfying extraction results for small size water body. The methods were evaluated by both overall accuracy (OAA) and Kappa coefficient (KC). The OAA of PCA-SVM, SVM, FNDWI-DTC, PCA are 96.68%, 94.23%, 93.99%, 93.01%, and the KCs are 0.9308, 0.8995, 0.8962, 0.8842, respectively, in consistent with visual inspection. In summary, SVM is better for narrow rivers extraction and PCA-SVM is suitable for water extraction of various types. As for dark blue lakes, the methods using PCA can extract more quickly and accurately.

  10. Manual of analytical methods for the Environmental Health Laboratory

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

    Gray, C. E.

    1975-06-01

    This manual was compiled from techniques used In the Environmental Health Laboratory of Sandia Laboratories at Albuquerque. New Mexico, and is a revision of an earlier publication (SC-M-07-3044) edited by Lial W. Brewer. The procedures arc similar to those used in other laboratories devoted to Environmental Health practices. Some of the methods are standard and others are modified to suit our needs; others were developed at Sandia. The author has attempted to present all methods in a simple and concise manner, but in sufficient detail to make them readily usable. It is not inferred that the methods are universal formore » any type of sample, but they have been found very reliable for the types of samples mentioned. The author will welcome inquiry for clarification of any part of this manual. It is the desire of the author that this manual will be of use and service to others. New and revised procedures will be issued as supplements to this document.« less

  11. Effects of extraction methods on the antioxidant activities of polysaccharides from Agaricus blazei Murrill.

    PubMed

    Jia, Shaoyi; Li, Feng; Liu, Yong; Ren, Haitao; Gong, Guili; Wang, Yanyan; Wu, Songhai

    2013-11-01

    Five polysaccharides were obtained from Agaricus blazei Murrill (ABM) through different extraction methods including hot water extraction, single enzyme extraction (pectinase, cellulase or papain) and compound enzymes extraction (cellulase:pectinase:papain). Their characteristics such as the polysaccharide yield, polysaccharide content, protein content, infrared spectra were determined, and antioxidant activities were investigated on the basis of hydroxyl radical, DPPH free radical, ABTS free radical and reducing power. The results showed that five extracts exhibited antioxidant activities in a concentration-dependent manner. Compared with other methods, the compound enzymes extraction method was found to present the highest polysaccharides yield (17.44%). Moreover, compound enzymes extracts exhibited the strongest reducing power and highest scavenging rates on hydroxyl radicals, DPPH radicals and ABTS radicals. On the contrary, hot water extraction method had the lowest polysaccharides yield of 11.95%, whose extracts also exhibited the lowest antioxidant activities. Overall, the available data obtained in vitro models suggested that ABM extracts were natural antioxidants and compound enzymes extraction was an appropriate, mild and effective extracting method for obtaining the polysaccharide extracts from Agaricus blazei Murrill (ABM). Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Introducing automation to the molecular diagnosis of Trypanosoma cruzi infection: A comparative study of sample treatments, DNA extraction methods and real-time PCR assays.

    PubMed

    Abras, Alba; Ballart, Cristina; Llovet, Teresa; Roig, Carme; Gutiérrez, Cristina; Tebar, Silvia; Berenguer, Pere; Pinazo, María-Jesús; Posada, Elizabeth; Gascón, Joaquim; Schijman, Alejandro G; Gállego, Montserrat; Muñoz, Carmen

    2018-01-01

    Polymerase chain reaction (PCR) has become a useful tool for the diagnosis of Trypanosoma cruzi infection. The development of automated DNA extraction methodologies and PCR systems is an important step toward the standardization of protocols in routine diagnosis. To date, there are only two commercially available Real-Time PCR assays for the routine laboratory detection of T. cruzi DNA in clinical samples: TCRUZIDNA.CE (Diagnostic Bioprobes Srl) and RealCycler CHAG (Progenie Molecular). Our aim was to evaluate the RealCycler CHAG assay taking into account the whole process. We assessed the usefulness of an automated DNA extraction system based on magnetic particles (EZ1 Virus Mini Kit v2.0, Qiagen) combined with a commercially available Real-Time PCR assay targeting satellite DNA (SatDNA) of T. cruzi (RealCycler CHAG), a methodology used for routine diagnosis in our hospital. It was compared with a well-known strategy combining a commercial DNA isolation kit based on silica columns (High Pure PCR Template Preparation Kit, Roche Diagnostics) with an in-house Real-Time PCR targeting SatDNA. The results of the two methodologies were in almost perfect agreement, indicating they can be used interchangeably. However, when variations in protocol factors were applied (sample treatment, extraction method and Real-Time PCR), the results were less convincing. A comprehensive fine-tuning of the whole procedure is the key to successful results. Guanidine EDTA-blood (GEB) samples are not suitable for DNA extraction based on magnetic particles due to inhibition, at least when samples are not processed immediately. This is the first study to evaluate the RealCycler CHAG assay taking into account the overall process, including three variables (sample treatment, extraction method and Real-Time PCR). Our findings may contribute to the harmonization of protocols between laboratories and to a wider application of Real-Time PCR in molecular diagnostic laboratories associated with health

  13. Laboratory based instruction in Pakistan: Comparative evaluation of three laboratory instruction methods in biological science at higher secondary school level

    NASA Astrophysics Data System (ADS)

    Cheema, Tabinda Shahid

    This study of laboratory based instruction at higher secondary school level was an attempt to gain some insight into the effectiveness of three laboratory instruction methods: cooperative group instruction method, individualised instruction method and lecture demonstration method on biology achievement and retention. A Randomised subjects, Pre-test Post-test Comparative Methods Design was applied. Three groups of students from a year 11 class in Pakistan conducted experiments using the different laboratory instruction methods. Pre-tests, achievement tests after the experiments and retention tests one month later were administered. Results showed no significant difference between the groups on total achievement and retention, nor was there any significant difference on knowledge and comprehension test scores or skills performance. Future research investigating a similar problem is suggested.

  14. Comprehensive GMO detection using real-time PCR array: single-laboratory validation.

    PubMed

    Mano, Junichi; Harada, Mioko; Takabatake, Reona; Furui, Satoshi; Kitta, Kazumi; Nakamura, Kosuke; Akiyama, Hiroshi; Teshima, Reiko; Noritake, Hiromichi; Hatano, Shuko; Futo, Satoshi; Minegishi, Yasutaka; Iizuka, Tayoshi

    2012-01-01

    We have developed a real-time PCR array method to comprehensively detect genetically modified (GM) organisms. In the method, genomic DNA extracted from an agricultural product is analyzed using various qualitative real-time PCR assays on a 96-well PCR plate, targeting for individual GM events, recombinant DNA (r-DNA) segments, taxon-specific DNAs, and donor organisms of the respective r-DNAs. In this article, we report the single-laboratory validation of both DNA extraction methods and component PCR assays constituting the real-time PCR array. We selected some DNA extraction methods for specified plant matrixes, i.e., maize flour, soybean flour, and ground canola seeds, then evaluated the DNA quantity, DNA fragmentation, and PCR inhibition of the resultant DNA extracts. For the component PCR assays, we evaluated the specificity and LOD. All DNA extraction methods and component PCR assays satisfied the criteria set on the basis of previous reports.

  15. New method to enhance the extraction yield of rutin from Sophora japonica using a novel ultrasonic extraction system by determining optimum ultrasonic frequency.

    PubMed

    Liao, Jianqing; Qu, Baida; Liu, Da; Zheng, Naiqin

    2015-11-01

    A new method has been proposed for enhancing extraction yield of rutin from Sophora japonica, in which a novel ultrasonic extraction system has been developed to perform the determination of optimum ultrasonic frequency by a two-step procedure. This study has systematically investigated the influence of a continuous frequency range of 20-92 kHz on rutin yields. The effects of different operating conditions on rutin yields have also been studied in detail such as solvent concentration, solvent to solid ratio, ultrasound power, temperature and particle size. A higher extraction yield was obtained at the ultrasonic frequency of 60-62 kHz which was little affected under other extraction conditions. Comparative studies between existing methods and the present method were done to verify the effectiveness of this method. Results indicated that the new extraction method gave a higher extraction yield compared with existing ultrasound-assisted extraction (UAE) and soxhlet extraction (SE). Thus, the potential use of this method may be promising for extraction of natural materials on an industrial scale in the future. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Valid methods: the quality assurance of test method development, validation, approval, and transfer for veterinary testing laboratories.

    PubMed

    Wiegers, Ann L

    2003-07-01

    Third-party accreditation is a valuable tool to demonstrate a laboratory's competence to conduct testing. Accreditation, internationally and in the United States, has been discussed previously. However, accreditation is only I part of establishing data credibility. A validated test method is the first component of a valid measurement system. Validation is defined as confirmation by examination and the provision of objective evidence that the particular requirements for a specific intended use are fulfilled. The international and national standard ISO/IEC 17025 recognizes the importance of validated methods and requires that laboratory-developed methods or methods adopted by the laboratory be appropriate for the intended use. Validated methods are therefore required and their use agreed to by the client (i.e., end users of the test results such as veterinarians, animal health programs, and owners). ISO/IEC 17025 also requires that the introduction of methods developed by the laboratory for its own use be a planned activity conducted by qualified personnel with adequate resources. This article discusses considerations and recommendations for the conduct of veterinary diagnostic test method development, validation, evaluation, approval, and transfer to the user laboratory in the ISO/IEC 17025 environment. These recommendations are based on those of nationally and internationally accepted standards and guidelines, as well as those of reputable and experienced technical bodies. They are also based on the author's experience in the evaluation of method development and transfer projects, validation data, and the implementation of quality management systems in the area of method development.

  17. [Comparison of minimally invasive extraction and traditional method in the extraction of impacted mandibular third molar].

    PubMed

    Xu, Fang; Zhang, Hui-Xia

    2016-10-01

    To compare minimally invasive extraction and traditional method in the extraction of impacted mandibular third molar. One hundred and sixty patients with impacted mandibular third molar were equally divided into two groups. Patients in the experimental group were treated with minimally invasive extraction, using implant machine and luxator, while patients in the control group were treated with traditional methods including use of orthodox chisel. The operation time, intraoperative and postoperative complications including deformation of extraction sockets, dry socket, limitation of mouth opening, pain and swelling, and fear were observed and compared between the two groups. The data were analyzed with SPSS18.0 software package. The operation time was (17.32±1.01) min in the experimental group, significantly shorter than the control group which was (33.46±1.12)min (P<0.05); significant difference was found in the incidence of root fracture, medium or severe tooth sockets deformation and incidence of psychological fear during operation between the control group and experimental group(P<0.05); the degree of mouth opening after surgery, the incidence of moderate or severe pain after surgery was significantly lower in the experimental than in the control group(P<0.05). Minimally invasive extraction of mandibular impacted wisdom tooth is better than traditional method, with shorter operation time and less intraoperative and postoperative complications, which should be widely applied in clinic.

  18. Sediment laboratory quality-assurance project: studies of methods and materials

    USGS Publications Warehouse

    Gordon, J.D.; Newland, C.A.; Gray, J.R.

    2001-01-01

    In August 1996 the U.S. Geological Survey initiated the Sediment Laboratory Quality-Assurance project. The Sediment Laboratory Quality Assurance project is part of the National Sediment Laboratory Quality-Assurance program. This paper addresses the fmdings of the sand/fme separation analysis completed for the single-blind reference sediment-sample project and differences in reported results between two different analytical procedures. From the results it is evident that an incomplete separation of fme- and sand-size material commonly occurs resulting in the classification of some of the fme-size material as sand-size material. Electron microscopy analysis supported the hypothesis that the negative bias for fme-size material and the positive bias for sand-size material is largely due to aggregation of some of the fine-size material into sand-size particles and adherence of fine-size material to the sand-size grains. Electron microscopy analysis showed that preserved river water, which was low in dissolved solids, specific conductance, and neutral pH, showed less aggregation and adhesion than preserved river water that was higher in dissolved solids and specific conductance with a basic pH. Bacteria were also found growing in the matrix, which may enhance fme-size material aggregation through their adhesive properties. Differences between sediment-analysis methods were also investigated as pan of this study. Suspended-sediment concentration results obtained from one participating laboratory that used a total-suspended solids (TSS) method had greater variability and larger negative biases than results obtained when this laboratory used a suspended-sediment concentration method. When TSS methods were used to analyze the reference samples, the median suspended sediment concentration percent difference was -18.04 percent. When the laboratory used a suspended-sediment concentration method, the median suspended-sediment concentration percent difference was -2

  19. DO TIE LABORATORY BASED METHODS REALLY REFLECT FIELD CONDITIONS

    EPA Science Inventory

    Sediment Toxicity Identification and Evaluation (TIE) methods have been developed for both interstitial waters and whole sediments. These relatively simple laboratory methods are designed to identify specific toxicants or classes of toxicants in sediments; however, the question ...

  20. Laboratory-based versus non-laboratory-based method for assessment of cardiovascular disease risk: the NHANES I Follow-up Study cohort

    PubMed Central

    Gaziano, Thomas A; Young, Cynthia R; Fitzmaurice, Garrett; Atwood, Sidney; Gaziano, J Michael

    2008-01-01

    Summary Background Around 80% of all cardiovascular deaths occur in developing countries. Assessment of those patients at high risk is an important strategy for prevention. Since developing countries have limited resources for prevention strategies that require laboratory testing, we assessed if a risk prediction method that did not require any laboratory tests could be as accurate as one requiring laboratory information. Methods The National Health and Nutrition Examination Survey (NHANES) was a prospective cohort study of 14 407 US participants aged between 25–74 years at the time they were first examined (between 1971 and 1975). Our follow-up study population included participants with complete information on these surveys who did not report a history of cardiovascular disease (myocardial infarction, heart failure, stroke, angina) or cancer, yielding an analysis dataset N=6186. We compared how well either method could predict first-time fatal and non-fatal cardiovascular disease events in this cohort. For the laboratory-based model, which required blood testing, we used standard risk factors to assess risk of cardiovascular disease: age, systolic blood pressure, smoking status, total cholesterol, reported diabetes status, and current treatment for hypertension. For the non-laboratory-based model, we substituted body-mass index for cholesterol. Findings In the cohort of 6186, there were 1529 first-time cardiovascular events and 578 (38%) deaths due to cardiovascular disease over 21 years. In women, the laboratory-based model was useful for predicting events, with a c statistic of 0·829. The c statistic of the non-laboratory-based model was 0·831. In men, the results were similar (0·784 for the laboratory-based model and 0·783 for the non-laboratory-based model). Results were similar between the laboratory-based and non-laboratory-based models in both men and women when restricted to fatal events only. Interpretation A method that uses non-laboratory

  1. Text-in-Context: A Method for Extracting Findings in Mixed-Methods Mixed Research Synthesis Studies

    PubMed Central

    Leeman, Jennifer; Knafl, Kathleen; Crandell, Jamie L.

    2012-01-01

    Aim Our purpose in this paper is to propose a new method for extracting findings from research reports included in mixed-methods mixed research synthesis studies. Background International initiatives in the domains of systematic review and evidence synthesis have been focused on broadening the conceptualization of evidence, increased methodological inclusiveness and the production of evidence syntheses that will be accessible to and usable by a wider range of consumers. Initiatives in the general mixed-methods research field have been focused on developing truly integrative approaches to data analysis and interpretation. Data source The data extraction challenges described here were encountered and the method proposed for addressing these challenges was developed, in the first year of the ongoing (2011–2016) study: Mixed-Methods Synthesis of Research on Childhood Chronic Conditions and Family. Discussion To preserve the text-in-context of findings in research reports, we describe a method whereby findings are transformed into portable statements that anchor results to relevant information about sample, source of information, time, comparative reference point, magnitude and significance and study-specific conceptions of phenomena. Implications for nursing The data extraction method featured here was developed specifically to accommodate mixed-methods mixed research synthesis studies conducted in nursing and other health sciences, but reviewers might find it useful in other kinds of research synthesis studies. Conclusion This data extraction method itself constitutes a type of integration to preserve the methodological context of findings when statements are read individually and in comparison to each other. PMID:22924808

  2. Methods for extracting social network data from chatroom logs

    NASA Astrophysics Data System (ADS)

    Osesina, O. Isaac; McIntire, John P.; Havig, Paul R.; Geiselman, Eric E.; Bartley, Cecilia; Tudoreanu, M. Eduard

    2012-06-01

    Identifying social network (SN) links within computer-mediated communication platforms without explicit relations among users poses challenges to researchers. Our research aims to extract SN links in internet chat with multiple users engaging in synchronous overlapping conversations all displayed in a single stream. We approached this problem using three methods which build on previous research. Response-time analysis builds on temporal proximity of chat messages; word context usage builds on keywords analysis and direct addressing which infers links by identifying the intended message recipient from the screen name (nickname) referenced in the message [1]. Our analysis of word usage within the chat stream also provides contexts for the extracted SN links. To test the capability of our methods, we used publicly available data from Internet Relay Chat (IRC), a real-time computer-mediated communication (CMC) tool used by millions of people around the world. The extraction performances of individual methods and their hybrids were assessed relative to a ground truth (determined a priori via manual scoring).

  3. PCR-fingerprint profiles of mitochondrial and genomic DNA extracted from Fetus cervi using different extraction methods.

    PubMed

    Ai, Jinxia; Wang, Xuesong; Gao, Lijun; Xia, Wei; Li, Mingcheng; Yuan, Guangxin; Niu, Jiamu; Zhang, Lihua

    2017-11-01

    The use of Fetus cervi, which is derived from the embryo and placenta of Cervus Nippon Temminck or Cervs elaphus Linnaeus, has been documented for a long time in China. There are abundant species of deer worldwide. Those recorded by China Pharmacopeia (2010 edition) from all the species were either authentic or adulterants/counterfeits. Identification of their origins or authenticity became a key in the preparation of the authentic products. The traditional SDS alkaline lysis and salt-outing methods were modified to extract mt DNA and genomic DNA from fresh and dry Fetus cervi in addition to Fetus from false animals, respectively. A set of primers were designed by bioinformatics to target the intra-and inter-variation. The mt DNA and genomic DNA extracted from Fetus cervi using the two methods meet the requirement for authenticity. Extraction of mt DNA by SDS alkaline lysis is more practical and accurate than extraction of genomic DNA by salt-outing method. There were differences in length and number of segments amplified by PCR between mt DNA from authentic Fetus cervi and false animals Fetus. The distinctive PCR-fingerprint patterns can distinguish the Fetus cervi from adulterants and counterfeit animal Fetus.

  4. Pseudophasic extraction method for the separation of ultra-fine minerals

    DOEpatents

    Chaiko, David J.

    2002-01-01

    An improved aqueous-based extraction method for the separation and recovery of ultra-fine mineral particles. The process operates within the pseudophase region of the conventional aqueous biphasic extraction system where a low-molecular-weight, water soluble polymer alone is used in combination with a salt and operates within the pseudo-biphase regime of the conventional aqueous biphasic extraction system. A combination of low molecular weight, mutually immiscible polymers are used with or without a salt. This method is especially suited for the purification of clays that are useful as rheological control agents and for the preparation of nanocomposites.

  5. A comparison of two microscale laboratory reporting methods in a secondary chemistry classroom

    NASA Astrophysics Data System (ADS)

    Martinez, Lance Michael

    This study attempted to determine if there was a difference between the laboratory achievement of students who used a modified reporting method and those who used traditional laboratory reporting. The study also determined the relationships between laboratory performance scores and the independent variables score on the Group Assessment of Logical Thinking (GALT) test, chronological age in months, gender, and ethnicity for each of the treatment groups. The study was conducted using 113 high school students who were enrolled in first-year general chemistry classes at Pueblo South High School in Colorado. The research design used was the quasi-experimental Nonequivalent Control Group Design. The statistical treatment consisted of the Multiple Regression Analysis and the Analysis of Covariance. Based on the GALT, students in the two groups were generally in the concrete and transitional stages of the Piagetian cognitive levels. The findings of the study revealed that the traditional and the modified methods of laboratory reporting did not have any effect on the laboratory performance outcome of the subjects. However, the students who used the traditional method of reporting showed a higher laboratory performance score when evaluation was conducted using the New Standards rubric recommended by the state. Multiple Regression Analysis revealed that there was a significant relationship between the criterion variable student laboratory performance outcome of individuals who employed traditional laboratory reporting methods and the composite set of predictor variables. On the contrary, there was no significant relationship between the criterion variable student laboratory performance outcome of individuals who employed modified laboratory reporting methods and the composite set of predictor variables.

  6. An Efficient Method for Genomic DNA Extraction from Different Molluscs Species

    PubMed Central

    Pereira, Jorge C.; Chaves, Raquel; Bastos, Estela; Leitão, Alexandra; Guedes-Pinto, Henrique

    2011-01-01

    The selection of a DNA extraction method is a critical step when subsequent analysis depends on the DNA quality and quantity. Unlike mammals, for which several capable DNA extraction methods have been developed, for molluscs the availability of optimized genomic DNA extraction protocols is clearly insufficient. Several aspects such as animal physiology, the type (e.g., adductor muscle or gills) or quantity of tissue, can explain the lack of efficiency (quality and yield) in molluscs genomic DNA extraction procedure. In an attempt to overcome these aspects, this work describes an efficient method for molluscs genomic DNA extraction that was tested in several species from different orders: Veneridae, Ostreidae, Anomiidae, Cardiidae (Bivalvia) and Muricidae (Gastropoda), with different weight sample tissues. The isolated DNA was of high molecular weight with high yield and purity, even with reduced quantities of tissue. Moreover, the genomic DNA isolated, demonstrated to be suitable for several downstream molecular techniques, such as PCR sequencing among others. PMID:22174651

  7. Method and apparatus for back-extracting metal chelates

    DOEpatents

    Wai, C.M.; Smart, N.G.; Lin, Y.

    1998-08-11

    A method is described for extracting metal and metalloid species from a solid or liquid substrate using a supercritical fluid solvent containing one or more chelating agents followed by back-extracting the metal and metalloid species from the metal and metalloid chelates formed thereby. The back-extraction acidic solution is performed utilizing an acidic solution. Upon sufficient exposure of the metal and metalloid chelates to the acidic solution, the metal and metalloid species are released from the chelates into the acid solution, while the chelating agent remains in the supercritical fluid solvent. The chelating agent is thereby regenerated and the metal and metalloid species recovered. 3 figs.

  8. [Comparison study of different methods for extracting volatile oil from bergamot].

    PubMed

    Chen, Fei; Li, Qun-li; Sheng, Liu-qing; Qiu, Jiao-ying

    2008-08-01

    To test different methods for extracting volatile oil from bergamot. The determination of bergapten was carried out by RP-HPLC. Four different ways of organic solvent extraction, steam-input distillation, distillation of the material mixed with water and press extraction were compared. Bergapten wasnt extracted by ways of steam-input distillation and distillation of the material mixed with water. The steam distillation extraction can be taken to extract volatile oil from bergamot for protecting humans' skins.

  9. Extraction and [superscript 1]H NMR Analysis of Fats from Convenience Foods: A Laboratory Experiment for Organic Chemistry

    ERIC Educational Resources Information Center

    Hartel, Aaron M.; Moore, Amy C.

    2014-01-01

    The extraction and analysis of fats from convenience foods (crackers, cookies, chips, candies) has been developed as an experiment for a second-year undergraduate organic chemistry laboratory course. Students gravimetrically determine the fat content per serving and then perform a [superscript 1]H NMR analysis of the recovered fat to determine the…

  10. Guaiacol Peroxidase Zymography for the Undergraduate Laboratory

    ERIC Educational Resources Information Center

    Wilkesman, Jeff; Castro, Diana; Contreras, Lellys M.; Kurz, Liliana

    2014-01-01

    This laboratory exercise presents a novel way to introduce undergraduate students to the specific detection of enzymatic activity by electrophoresis. First, students prepare a crude peroxidase extract and then analyze the homogenate via electrophoresis. Zymography, that is, a SDS-PAGE method to detect enzyme activity, is used to specifically…

  11. Comparison of DNA extraction methods for human gut microbial community profiling.

    PubMed

    Lim, Mi Young; Song, Eun-Ji; Kim, Sang Ho; Lee, Jangwon; Nam, Young-Do

    2018-03-01

    The human gut harbors a vast range of microbes that have significant impact on health and disease. Therefore, gut microbiome profiling holds promise for use in early diagnosis and precision medicine development. Accurate profiling of the highly complex gut microbiome requires DNA extraction methods that provide sufficient coverage of the original community as well as adequate quality and quantity. We tested nine different DNA extraction methods using three commercial kits (TianLong Stool DNA/RNA Extraction Kit (TS), QIAamp DNA Stool Mini Kit (QS), and QIAamp PowerFecal DNA Kit (QP)) with or without additional bead-beating step using manual or automated methods and compared them in terms of DNA extraction ability from human fecal sample. All methods produced DNA in sufficient concentration and quality for use in sequencing, and the samples were clustered according to the DNA extraction method. Inclusion of bead-beating step especially resulted in higher degrees of microbial diversity and had the greatest effect on gut microbiome composition. Among the samples subjected to bead-beating method, TS kit samples were more similar to QP kit samples than QS kit samples. Our results emphasize the importance of mechanical disruption step for a more comprehensive profiling of the human gut microbiome. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

  12. An effective hand vein feature extraction method.

    PubMed

    Li, Haigang; Zhang, Qian; Li, Chengdong

    2015-01-01

    As a new authentication method developed years ago, vein recognition technology features the unique advantage of bioassay. This paper studies the specific procedure for the extraction of hand back vein characteristics. There are different positions used in the collecting process, so that a suitable intravenous regional orientation method is put forward, allowing the positioning area to be the same for all hand positions. In addition, to eliminate the pseudo vein area, the valley regional shape extraction operator can be improved and combined with multiple segmentation algorithms. The images should be segmented step by step, making the vein texture to appear clear and accurate. Lastly, the segmented images should be filtered, eroded, and refined. This process helps to filter the most of the pseudo vein information. Finally, a clear vein skeleton diagram is obtained, demonstrating the effectiveness of the algorithm. This paper presents a hand back vein region location method. This makes it possible to rotate and correct the image by working out the inclination degree of contour at the side of hand back.

  13. Evaluation of the essential oil of Foeniculum vulgare Mill (fennel) fruits extracted by three different extraction methods by GC/MS.

    PubMed

    Hammouda, Faiza M; Saleh, Mahmoud A; Abdel-Azim, Nahla S; Shams, Khaled A; Ismail, Shams I; Shahat, Abdelaaty A; Saleh, Ibrahim A

    2014-01-01

    Hydrodistillation (HD) and steam-distillation, or solvent extraction methods of essential oils have some disadvantages like thermal decomposition of extracts, its contamination with solvent or solvent residues and the pollution of residual vegetal material with solvent which can be also an environmental problem. Thus, new green techniques, such as supercritical fluid extraction and microwave assisted techniques, are potential solutions to overcome these disadvantages. The aim of this study was to evaluate the essential oil of Foeniculum vulgare subsp. Piperitum fruits extracted by three different extraction methods viz. Supercritical fluid extraction (SFE) using CO2, microwave-assisted extraction (MAE) and hydro-distillation (HD) using gas chromatography-mass spectrometry (GC/MS). The results revealed that both MAE and SFE enhanced the extraction efficiency of the interested components. MAE gave the highest yield of oil as well as higher percentage of Fenchone (28%), whereas SFE gave the highest percentage of anethol (72%). Microwave-assisted extraction (MAE) and supercritical fluid extraction (SFE) not only enhanced the essential oil extraction but also saved time, reduced the solvents use and produced, ecologically, green technologies.

  14. Performance testing of NIOSH Method 5524/ASTM Method D-7049-04, for determination of metalworking fluids.

    PubMed

    Glaser, Robert; Kurimo, Robert; Shulman, Stanley

    2007-08-01

    A performance test of NIOSH Method 5524/ASTM Method D-7049-04 for analysis of metalworking fluids (MWF) was conducted. These methods involve determination of the total and extractable weights of MWF samples; extractions are performed using a ternary blend of toluene:dichloromethane:methanol and a binary blend of methanol:water. Six laboratories participated in this study. A preliminary analysis of 20 blank samples was made to familiarize the laboratories with the procedure(s) and to estimate the methods' limits of detection/quantitation (LODs/LOQs). Synthetically generated samples of a semisynthetic MWF aerosol were then collected on tared polytetrafluoroethylene (PTFE) filters and analyzed according to the methods by all participants. Sample masses deposited (approximately 400-500 micro g) corresponded to amounts expected in an 8-hr shift at the NIOSH recommended exposure levels (REL) of 0.4 mg/m(3) (thoracic) and 0.5 mg/m(3) (total particulate). The generator output was monitored with a calibrated laser particle counter. One laboratory significantly underreported the sampled masses relative to the other five labs. A follow-up study compared only gravimetric results of this laboratory with those of two other labs. In the preliminary analysis of blanks; the average LOQs were 0.094 mg for the total weight analysis and 0.136 mg for the extracted weight analyses. For the six-lab study, the average LOQs were 0.064 mg for the total weight analyses and 0.067 mg for the extracted weight analyses. Using ASTM conventions, h and k statistics were computed to determine the degree of consistency of each laboratory with the others. One laboratory experienced problems with precision but not bias. The precision estimates for the remaining five labs were not different statistically (alpha = 0.005) for either the total or extractable weights. For all six labs, the average fraction extracted was > or =0.94 (CV = 0.025). Pooled estimates of the total coefficients of variation of

  15. Research of facial feature extraction based on MMC

    NASA Astrophysics Data System (ADS)

    Xue, Donglin; Zhao, Jiufen; Tang, Qinhong; Shi, Shaokun

    2017-07-01

    Based on the maximum margin criterion (MMC), a new algorithm of statistically uncorrelated optimal discriminant vectors and a new algorithm of orthogonal optimal discriminant vectors for feature extraction were proposed. The purpose of the maximum margin criterion is to maximize the inter-class scatter while simultaneously minimizing the intra-class scatter after the projection. Compared with original MMC method and principal component analysis (PCA) method, the proposed methods are better in terms of reducing or eliminating the statistically correlation between features and improving recognition rate. The experiment results on Olivetti Research Laboratory (ORL) face database shows that the new feature extraction method of statistically uncorrelated maximum margin criterion (SUMMC) are better in terms of recognition rate and stability. Besides, the relations between maximum margin criterion and Fisher criterion for feature extraction were revealed.

  16. An automatic rat brain extraction method based on a deformable surface model.

    PubMed

    Li, Jiehua; Liu, Xiaofeng; Zhuo, Jiachen; Gullapalli, Rao P; Zara, Jason M

    2013-08-15

    The extraction of the brain from the skull in medical images is a necessary first step before image registration or segmentation. While pre-clinical MR imaging studies on small animals, such as rats, are increasing, fully automatic imaging processing techniques specific to small animal studies remain lacking. In this paper, we present an automatic rat brain extraction method, the Rat Brain Deformable model method (RBD), which adapts the popular human brain extraction tool (BET) through the incorporation of information on the brain geometry and MR image characteristics of the rat brain. The robustness of the method was demonstrated on T2-weighted MR images of 64 rats and compared with other brain extraction methods (BET, PCNN, PCNN-3D). The results demonstrate that RBD reliably extracts the rat brain with high accuracy (>92% volume overlap) and is robust against signal inhomogeneity in the images. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. A multiple distributed representation method based on neural network for biomedical event extraction.

    PubMed

    Wang, Anran; Wang, Jian; Lin, Hongfei; Zhang, Jianhai; Yang, Zhihao; Xu, Kan

    2017-12-20

    Biomedical event extraction is one of the most frontier domains in biomedical research. The two main subtasks of biomedical event extraction are trigger identification and arguments detection which can both be considered as classification problems. However, traditional state-of-the-art methods are based on support vector machine (SVM) with massive manually designed one-hot represented features, which require enormous work but lack semantic relation among words. In this paper, we propose a multiple distributed representation method for biomedical event extraction. The method combines context consisting of dependency-based word embedding, and task-based features represented in a distributed way as the input of deep learning models to train deep learning models. Finally, we used softmax classifier to label the example candidates. The experimental results on Multi-Level Event Extraction (MLEE) corpus show higher F-scores of 77.97% in trigger identification and 58.31% in overall compared to the state-of-the-art SVM method. Our distributed representation method for biomedical event extraction avoids the problems of semantic gap and dimension disaster from traditional one-hot representation methods. The promising results demonstrate that our proposed method is effective for biomedical event extraction.

  18. Methods for producing extracted and digested products from pretreated lignocellulosic biomass

    DOEpatents

    Chundawat, Shishir; Sousa, Leonardo Da Costa; Cheh, Albert M.; Balan; , Venkatesh; Dale, Bruce

    2017-05-16

    Methods for producing extracted and digested products from pretreated lignocellulosic biomass are provided. The methods include converting native cellulose I.sub..beta. to cellulose III.sub.I by pretreating the lignocellulosic biomass with liquid ammonia under certain conditions, and performing extracting or digesting steps on the pretreated/converted lignocellulosic biomass.

  19. A new method to extract modal parameters using output-only responses

    NASA Astrophysics Data System (ADS)

    Kim, Byeong Hwa; Stubbs, Norris; Park, Taehyo

    2005-04-01

    This work proposes a new output-only modal analysis method to extract mode shapes and natural frequencies of a structure. The proposed method is based on an approach with a single-degree-of-freedom in the time domain. For a set of given mode-isolated signals, the un-damped mode shapes are extracted utilizing the singular value decomposition of the output energy correlation matrix with respect to sensor locations. The natural frequencies are extracted from a noise-free signal that is projected on the estimated modal basis. The proposed method is particularly efficient when a high resolution of mode shape is essential. The accuracy of the method is numerically verified using a set of time histories that are simulated using a finite-element method. The feasibility and practicality of the method are verified using experimental data collected at the newly constructed King Storm Water Bridge in California, United States.

  20. A simplified method for extracting androgens from avian egg yolks

    USGS Publications Warehouse

    Kozlowski, C.P.; Bauman, J.E.; Hahn, D.C.

    2009-01-01

    Female birds deposit significant amounts of steroid hormones into the yolks of their eggs. Studies have demonstrated that these hormones, particularly androgens, affect nestling growth and development. In order to measure androgen concentrations in avian egg yolks, most authors follow the extraction methods outlined by Schwabl (1993. Proc. Nat. Acad. Sci. USA 90:11446-11450). We describe a simplified method for extracting androgens from avian egg yolks. Our method, which has been validated through recovery and linearity experiments, consists of a single ethanol precipitation that produces substantially higher recoveries than those reported by Schwabl.

  1. Comparison of four DNA extraction methods for the detection of Mycobacterium leprae from Ziehl-Neelsen-stained microscopic slides.

    PubMed

    Ruiz-Fuentes, Jenny Laura; Díaz, Alexis; Entenza, Anayma Elena; Frión, Yahima; Suárez, Odelaisy; Torres, Pedro; de Armas, Yaxsier; Acosta, Lucrecia

    2015-12-01

    The diagnosis of leprosy has been a challenge due to the low sensibility of the conventional methods and the impossibility of culturing the causative organism. In this study, four methods for Mycobacterium leprae nucleic-acid extraction from Ziehl-Neelsen-stained slides (ZNS slides) were compared: Phenol/chloroform, Chelex 100 resin, and two commercial kits (Wizard Genomic DNA Purification Kit and QIAamp DNA Mini Kit). DNA was extracted from four groups of slides: a high-codification-slide group (bacteriological index [BI]⩾4), a low-codification-slide group (BI=1), a negative-slide group (BI=0), and a negative-control-slide group (BI=0). Quality DNA was evidenced by the amplification of specific repetitive element present in M. leprae genomic DNA (RLEP) using a nested polymerase chain reaction. This is the first report comparing four different extraction methods for obtaining M. leprae DNA from ZNS slides in Cuban patients, and applied in molecular diagnosis. Good-quality DNA and positive amplification were detected in the high-codification-slide group with the four methods, while from the low-codification-slide group only the QIAGEN and phenol-chloroform methods obtained amplification of M. leprae. In the negative-slide group, only the QIAGEN method was able to obtain DNA with sufficient quality for positive amplification of the RLEP region. No amplification was observed in the negative-control-slide group by any method. Patients with ZNS negative slides can still transmit the infection, and molecular methods can help identify and treat them, interrupting the chain of transmission and preventing the onset of disabilities. The ZNS slides can be sent easily to reference laboratories for later molecular analysis that can be useful not only to improve the diagnosis, but also for the application of other molecular techniques. Copyright © 2015 Asian-African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

  2. A cell extraction method for oily sediments

    NASA Astrophysics Data System (ADS)

    Lappé, M.; Kallmeyer, J.

    2012-04-01

    Hydrocarbons can be found in many different habitats and represent an important carbon source for microbes. As fossil fuels, they are an important economical resource and, through natural seepage or accidental release, they can be major pollutants. Oil sands from Alberta, Canada, and samples from the seafloor of the Gulf of Mexico represent typical examples of either natural or anthropogenically affected oily sediments. DNA-specific stains and molecular probes bind to hydrocarbons, causing massive background fluorescence and thereby massively hampering cell enumeration. The cell extraction procedure of Kallmeyer et al. (2008) separates the cells from the sediment matrix, producing a sediment free cell extract that can then be used for subsequent staining and cell enumeration under a fluorescence microscope. In principle, this technique can also be used to separate cells from oily sediments, but it was not originally optimized for this application and does not provide satisfactory results. Here we present a modified extraction method in which the hydrocarbons are removed prior to cell extraction by a solvent treatment. Due to the reduced background fluorescence the microscopic image becomes clearer, making cell identification and enumeration much easier. Consequently, the resulting cell counts from oily samples treated according to our new protocol were significantly higher than those treated according to Kallmeyer et al. (2008). We tested different amounts of a variety of solvents for their ability to remove hydrocarbons and found that n-hexane and - in samples containing more biodegraded oils - methanol, delivered the best results. Because solvents also tend to lyse cells, it was important to find the optimum solvent to sample ratio, at which the positive effect of hydrocarbon extraction overcomes the negative effect of cell lysis. A volumetric ratio of 1:2 to 1:5 between a formalin-fixed sediment slurry and solvent delivered highest cell counts. Extraction

  3. Anti-ulcer and ulcer healing potentials of Musa sapientum peel extract in the laboratory rodents.

    PubMed

    Onasanwo, Samuel Adetunji; Emikpe, Benjamin Obukowho; Ajah, Austin Azubuike; Elufioye, Taiwo Olayemi

    2013-07-01

    This study investigated the anti-ulcer and ulcer healing potentials of the methanol extract of Musa sapientum peel in the laboratory rats. Methanol extract of the peels on Musa sapientum (MEMS) was evaluated for its anti-ulcer using alcohol-induced, aspirin-induced, and pyloric ligation-induced models, and for its ulcer healing employing acetic acid-induced ulcer models in rats. The findings from this experiment showed that MEMS (50, 100 and 200 mg/kg, b.w.) anti-ulcer and ulcer healing activity (P ≤ 0.05) is dose-dependent. Also, MEMS exhibited healing of the ulcer base in all the treated groups when compared with the control group. The outcomes of this experiment revealed that the anti-ulcer effect of MEMS may be due to its anti-secretory and cyto-protective activity. The healing of the ulcer base might not be unconnected with basic fibroblast growth factors responsible for epithelial regeneration.

  4. Accelerated, microwave-assisted, and conventional solvent extraction methods affect anthocyanin composition from colored grains.

    PubMed

    Abdel-Aal, El-Sayed M; Akhtar, Humayoun; Rabalski, Iwona; Bryan, Michael

    2014-02-01

    Anthocyanins are important dietary components with diverse positive functions in human health. This study investigates effects of accelerated solvent extraction (ASE) and microwave-assisted extraction (MAE) on anthocyanin composition and extraction efficiency from blue wheat, purple corn, and black rice in comparison with the commonly used solvent extraction (CSE). Factorial experimental design was employed to study effects of ASE and MAE variables, and anthocyanin extracts were analyzed by spectrophotometry, high-performance liquid chromatography-diode array detector (DAD), and liquid chromatography-mass spectrometry chromatography. The extraction efficiency of ASE and MAE was comparable with CSE at the optimal conditions. The greatest extraction by ASE was achieved at 50 °C, 2500 psi, 10 min using 5 cycles, and 100% flush. For MAE, a combination of 70 °C, 300 W, and 10 min in MAE was the most effective in extracting anthocyanins from blue wheat and purple corn compared with 50 °C, 1200 W, and 20 min for black rice. The anthocyanin composition of grain extracts was influenced by the extraction method. The ASE extraction method seems to be more appropriate in extracting anthocyanins from the colored grains as being comparable with the CSE method based on changes in anthocyanin composition. The method caused lower structural changes in anthocaynins compared with the MAE method. Changes in blue wheat anthocyanins were lower in comparison with purple corn or black rice perhaps due to the absence of acylated anthocyanin compounds in blue wheat. The results show significant differences in anthocyanins among the 3 extraction methods, which indicate a need to standardize a method for valid comparisons among studies and for quality assurance purposes. © 2014 Her Majesty the Queen in Right of Canada Journal of Food Science © 2014 Institute of Food Technologists® Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada.

  5. Extraction kinetic modelling of total polyphenols and total anthocyanins from saffron floral bio-residues: Comparison of extraction methods.

    PubMed

    Da Porto, Carla; Natolino, Andrea

    2018-08-30

    Analysis of the extraction kinetic modelling for natural compounds is essential for industrial application. The second order rate model was applied to estimate the extraction kinetics of conventional solid-liquid extraction (CSLE), ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) of total polyphenols (TPC) from saffron floral bio-residues at different solid-to-liquid ratios (R S/L )(1:10, 1:20, 1:30, 1:50 g ml -1 ), ethanol 59% as solvent and 66 °C temperature. The optimum solid-to-liquid ratios for TPC kinetics were 1:20 for CLSE, 1:30 for UAE and 1:50 for MAE. The kinetics of total anthocyanins (TA) and antioxidant activity (AA) were investigated for the optimum R S/L for each method. The results showed a good prediction of the model for extraction kinetics in all experiments (R 2  > 0.99; NRMS 0.65-3.35%). The kinetic parameters were calculated and discussed. UAE, compared with the other methods, had the greater efficiency for TPC, TA and AA. Copyright © 2018 Elsevier Ltd. All rights reserved.

  6. Comparison of Instream and Laboratory Methods of Measuring Sediment Oxygen Demand

    USGS Publications Warehouse

    Hall, Dennis C.; Berkas, Wayne R.

    1988-01-01

    Sediment oxygen demand (SOD) was determined at three sites in a gravel-bottomed central Missouri stream by: (1) two variations of an instream method, and (2) a laboratory method. SOD generally was greatest by the instream methods, which are considered more accurate, and least by the laboratory method. Disturbing stream sediment did not significantly decrease SOD by the instream method. Temperature ranges of up to 12 degree Celsius had no significant effect on the SOD. In the gravel-bottomed stream, the placement of chambers was critical to obtain reliable measurements. SOD rates were dependent on the method; therefore, care should be taken in comparing SOD data obtained by different methods. There is a need for a carefully researched standardized method for SOD determinations.

  7. A new method for microwave assisted ethanolic extraction of Mentha rotundifolia bioactive terpenoids.

    PubMed

    García-Sarrió, María Jesús; Sanz, María Luz; Sanz, Jesús; González-Coloma, Azucena; Cristina Soria, Ana

    2018-04-14

    A new microwave-assisted extraction (MAE) method using ethanol as solvent has been optimized by means of a Box-Behnken experimental design for the enhanced extraction of bioactive terpenoids from Mentha rotundifolia leaves; 100°C, 5 min, 1.125 g dry sample: 10 mL solvent and a single extraction cycle were selected as optimal conditions. Improved performance of MAE method in terms of extraction yield and/or reproducibility over conventional solid-liquid extraction and ultrasound assisted extraction was also previously assessed. A comprehensive characterization of MAE extracts was carried out by GC-MS. A total of 46 compounds, mostly terpenoids, were identified; piperitenone oxide and piperitenone were the major compounds determined. Several neophytadiene isomers were also detected for the first time in MAE extracts. Different procedures (solid-phase extraction and activated charcoal (AC) treatment) were also evaluated for clean-up of MAE extracts, with AC providing the highest enrichment in bioactive terpenoids. Finally, the MAE method here developed is shown as a green, fast, efficient and reproducible liquid extraction methodology to obtain M. rotundifolia bioactive extracts for further application, among others, as food preservatives. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Exact extraction method for road rutting laser lines

    NASA Astrophysics Data System (ADS)

    Hong, Zhiming

    2018-02-01

    This paper analyzes the importance of asphalt pavement rutting detection in pavement maintenance and pavement administration in today's society, the shortcomings of the existing rutting detection methods are presented and a new rutting line-laser extraction method based on peak intensity characteristic and peak continuity is proposed. The intensity of peak characteristic is enhanced by a designed transverse mean filter, and an intensity map of peak characteristic based on peak intensity calculation for the whole road image is obtained to determine the seed point of the rutting laser line. Regarding the seed point as the starting point, the light-points of a rutting line-laser are extracted based on the features of peak continuity, which providing exact basic data for subsequent calculation of pavement rutting depths.

  9. Analysis of the essential oils of Alpiniae Officinarum Hance in different extraction methods

    NASA Astrophysics Data System (ADS)

    Yuan, Y.; Lin, L. J.; Huang, X. B.; Li, J. H.

    2017-09-01

    It was developed for the analysis of the essential oils of Alpiniae Officinarum Hance extracted by steam distillation (SD), ultrasonic assisted solvent extraction (UAE) and supercritical fluid extraction (SFE) via gas chromatography mass spectrometry (GC-MS) combined with retention index (RI) method. There were multiple volatile components of the oils extracted by the three above-mention methods respectively identified; meanwhile, each one was quantified by area normalization method. The results indicated that the content of 1,8-Cineole, the index constituent, by SD was similar as SFE, and higher than UAE. Although UAE was less time consuming and consumed less energy, the oil quality was poorer due to the use of organic solvents was hard to degrade. In addition, some constituents could be obtained by SFE but could not by SD. In conclusion, essential oil of different extraction methods from the same batch of materials had been proved broadly similarly, however, there were some differences in composition and component ratio. Therefore, development and utilization of different extraction methods must be selected according to the functional requirements of products.

  10. Safety in the Chemical Laboratory: Tested Disposal Methods for Chemical Wastes from Academic Laboratories.

    ERIC Educational Resources Information Center

    Armour, M. A.; And Others

    1985-01-01

    Describes procedures for disposing of dichromate cleaning solution, picric acid, organic azides, oxalic acid, chemical spills, and hydroperoxides in ethers and alkenes. These methods have been tested under laboratory conditions and are specific for individual chemicals rather than for groups of chemicals. (JN)

  11. Evaluation of asphalt cement extraction and recovery methods : final report.

    DOT National Transportation Integrated Search

    1982-07-01

    This study was concerned with the quality of recovered asphalt cement which may be attributable to the method used for extraction or to the Abson recovery. Variables of these procedures such as the time an asphalt cement was exposed to extraction sol...

  12. The Effect of Sericin from Various Extraction Methods on Cell Viability and Collagen Production

    PubMed Central

    Aramwit, Pornanong; Kanokpanont, Sorada; Nakpheng, Titpawan; Srichana, Teerapol

    2010-01-01

    Silk sericin (SS) can accelerate cell proliferation and attachment; however, SS can be extracted by various methods, which result in SS exhibiting different physical and biological properties. We found that SS produced from various extraction methods has different molecular weights, zeta potential, particle size and amino acid content. The MTT assay indicated that SS from all extraction methods had no toxicity to mouse fibroblast cells at concentrations up to 40 μg/mL after 24 h incubation, but SS obtained from some extraction methods can be toxic at higher concentrations. Heat-degraded SS was the least toxic to cells and activated the highest collagen production, while urea-extracted SS showed the lowest cell viability and collagen production. SS from urea extraction was severely harmful to cells at concentrations higher than 100 μg/mL. SS from all extraction methods could still promote collagen production in a concentration-dependent manner, even at high concentrations that are toxic to cells. PMID:20559510

  13. Evaluation of DNA extraction methods for the analysis of microbial community in biological activated carbon.

    PubMed

    Zheng, Lu; Gao, Naiyun; Deng, Yang

    2012-01-01

    It is difficult to isolate DNA from biological activated carbon (BAC) samples used in water treatment plants, owing to the scarcity of microorganisms in BAC samples. The aim of this study was to identify DNA extraction methods suitable for a long-term, comprehensive ecological analysis of BAC microbial communities. To identify a procedure that can produce high molecular weight DNA, maximizes detectable diversity and is relatively free from contaminants, the microwave extraction method, the cetyltrimethylammonium bromide (CTAB) extraction method, a commercial DNA extraction kit, and the ultrasonic extraction method were used for the extraction of DNA from BAC samples. Spectrophotometry, agarose gel electrophoresis and polymerase chain reaction (PCR)-restriction fragment length polymorphisms (RFLP) analysis were conducted to compare the yield and quality of DNA obtained using these methods. The results showed that the CTAB method produce the highest yield and genetic diversity of DNA from BAC samples, but DNA purity was slightly less than that obtained with the DNA extraction-kit method. This study provides a theoretical basis for establishing and selecting DNA extraction methods for BAC samples.

  14. Hot-Alkaline DNA Extraction Method for Deep-Subseafloor Archaeal Communities

    PubMed Central

    Terada, Takeshi; Hoshino, Tatsuhiko; Inagaki, Fumio

    2014-01-01

    A prerequisite for DNA-based microbial community analysis is even and effective cell disruption for DNA extraction. With a commonly used DNA extraction kit, roughly two-thirds of subseafloor sediment microbial cells remain intact on average (i.e., the cells are not disrupted), indicating that microbial community analyses may be biased at the DNA extraction step, prior to subsequent molecular analyses. To address this issue, we standardized a new DNA extraction method using alkaline treatment and heating. Upon treatment with 1 M NaOH at 98°C for 20 min, over 98% of microbial cells in subseafloor sediment samples collected at different depths were disrupted. However, DNA integrity tests showed that such strong alkaline and heat treatment also cleaved DNA molecules into short fragments that could not be amplified by PCR. Subsequently, we optimized the alkaline and temperature conditions to minimize DNA fragmentation and retain high cell disruption efficiency. The best conditions produced a cell disruption rate of 50 to 80% in subseafloor sediment samples from various depths and retained sufficient DNA integrity for amplification of the complete 16S rRNA gene (i.e., ∼1,500 bp). The optimized method also yielded higher DNA concentrations in all samples tested compared with extractions using a conventional kit-based approach. Comparative molecular analysis using real-time PCR and pyrosequencing of bacterial and archaeal 16S rRNA genes showed that the new method produced an increase in archaeal DNA and its diversity, suggesting that it provides better analytical coverage of subseafloor microbial communities than conventional methods. PMID:24441163

  15. The Extraction of Terrace in the Loess Plateau Based on radial method

    NASA Astrophysics Data System (ADS)

    Liu, W.; Li, F.

    2016-12-01

    The terrace of Loess Plateau, as a typical kind of artificial landform and an important measure of soil and water conservation, its positioning and automatic extraction will simplify the work of land use investigation. The existing methods of terrace extraction mainly include visual interpretation and automatic extraction. The manual method is used in land use investigation, but it is time-consuming and laborious. Researchers put forward some automatic extraction methods. For example, Fourier transform method can recognize terrace and find accurate position from frequency domain image, but it is more affected by the linear objects in the same direction of terrace; Texture analysis method is simple and have a wide range application of image processing. The disadvantage of texture analysis method is unable to recognize terraces' edge; Object-oriented is a new method of image classification, but when introduce it to terrace extracting, fracture polygons will be the most serious problem and it is difficult to explain its geological meaning. In order to positioning the terraces, we use high- resolution remote sensing image to extract and analyze the gray value of the pixels which the radial went through. During the recognition process, we firstly use the DEM data analysis or by manual selecting, to roughly confirm the position of peak points; secondly, take each of the peak points as the center to make radials in all directions; finally, extracting the gray values of the pixels which the radials went through, and analyzing its changing characteristics to confirm whether the terrace exists. For the purpose of getting accurate position of terrace, terraces' discontinuity, extension direction, ridge width, image processing algorithm, remote sensing image illumination and other influence factors were fully considered when designing the algorithms.

  16. Selection of an Appropriate Protein Extraction Method to Study the Phosphoproteome of Maize Photosynthetic Tissue

    PubMed Central

    Luís, Inês M.; Alexandre, Bruno M.; Oliveira, M. Margarida

    2016-01-01

    Often plant tissues are recalcitrant and, due to that, methods relying on protein precipitation, such as TCA/acetone precipitation and phenol extraction, are usually the methods of choice for protein extraction in plant proteomic studies. However, the addition of precipitation steps to protein extraction methods may negatively impact protein recovery, due to problems associated with protein re-solubilization. Moreover, we show that when working with non-recalcitrant plant tissues, such as young maize leaves, protein extraction methods with precipitation steps compromise the maintenance of some labile post-translational modifications (PTMs), such as phosphorylation. Therefore, a critical issue when studying PTMs in plant proteins is to ensure that the protein extraction method is the most appropriate, both at qualitative and quantitative levels. In this work, we compared five methods for protein extraction of the C4-photosynthesis related proteins, in the tip of fully expanded third-leaves. These included: TCA/Acetone Precipitation; Phenol Extraction; TCA/Acetone Precipitation followed by Phenol Extraction; direct extraction in Lysis Buffer (a urea-based buffer); and direct extraction in Lysis Buffer followed by Cleanup with a commercial kit. Protein extraction in Lysis Buffer performed better in comparison to the other methods. It gave one of the highest protein yields, good coverage of the extracted proteome and phosphoproteome, high reproducibility, and little protein degradation. This was also the easiest and fastest method, warranting minimal sample handling. We also show that this method is adequate for the successful extraction of key enzymes of the C4-photosynthetic metabolism, such as PEPC, PPDK, PEPCK, and NADP-ME. This was confirmed by MALDI-TOF/TOF MS analysis of excised spots of 2DE analyses of the extracted protein pools. Staining for phosphorylated proteins in 2DE revealed the presence of several phosphorylated isoforms of PEPC, PPDK, and PEPCK. PMID

  17. [How to Interpret and Use Routine Laboratory Data--Our Methods to Interpret Routine Laboratory Data--Chairmen's Introductory Remarks].

    PubMed

    Honda, Takayuki; Tozuka, Minoru

    2015-09-01

    In the reversed clinicopathological conference (R-CPC), three specialists in laboratory medicine interpreted routine laboratory data independently in order to understand the detailed state of a patient. R-CPC is an educational method to use laboratory data appropriately, and it is also important to select differential diagnoses in a process of clinical reasoning in addition to the present illness and physical examination. Routine laboratory tests can be performed repeatedly at a relatively low cost, and their time-series analysis can be performed. Interpretation of routine laboratory data is almost the same as taking physical findings. General findings are initially checked and then the state of each organ is examined. Although routine laboratory tests cost little, we can gain much more information from them about the patient than physical examinations.

  18. Comparative Efficiency of the Fenwick Can and Schuiling Centrifuge in Extracting Nematode Cysts from Different Soil Types

    PubMed Central

    Bellvert, Joaquim; Crombie, Kieran; Horgan, Finbarr G.

    2008-01-01

    The Fenwick can and Schuiling centrifuge are widely used to extract nematode cysts from soil samples. The comparative efficiencies of these two methods during cyst extraction have not been determined for different soil types under different cyst densities. Such information is vital for statutory laboratories that must choose a method for routine, high-throughput soil monitoring. In this study, samples of different soil types seeded with varying densities of potato cyst nematode (Globodera rostochiensis) cysts were processed using both methods. In one experiment, with 200 ml samples, recovery was similar between methods. In a second experiment with 500 ml samples, cyst recovery was higher using the Schuiling centrifuge. For each method and soil type, cyst extraction efficiency was similar across all densities tested. Extraction was efficient from pure sand (Fenwick 72%, Schuiling 84%) and naturally sandy soils (Fenwick 62%, Schuiling 73%), but was significantly less efficient from clay-soil (Fenwick 42%, Schuiling 44%) and peat-soil with high organic matter content (Fenwick 35%, Schuiling 33%). Residual moisture (<10% w/w) in samples prior to analyses reduced extraction efficiency, particularly for sand and sandy soils. For each soil type and method, there were significant linear relationships between the number of cysts extracted and the numbers of cysts in the samples. We discuss the advantages and disadvantages of each extraction method for cyst extraction in statutory soil laboratories. PMID:19259516

  19. Comparison of a New Cobinamide-Based Method to a Standard Laboratory Method for Measuring Cyanide in Human Blood

    PubMed Central

    Swezey, Robert; Shinn, Walter; Green, Carol; Drover, David R.; Hammer, Gregory B.; Schulman, Scott R.; Zajicek, Anne; Jett, David A.; Boss, Gerry R.

    2013-01-01

    Most hospital laboratories do not measure blood cyanide concentrations, and samples must be sent to reference laboratories. A simple method is needed for measuring cyanide in hospitals. The authors previously developed a method to quantify cyanide based on the high binding affinity of the vitamin B12 analog, cobinamide, for cyanide and a major spectral change observed for cyanide-bound cobinamide. This method is now validated in human blood, and the findings include a mean inter-assay accuracy of 99.1%, precision of 8.75% and a lower limit of quantification of 3.27 µM cyanide. The method was applied to blood samples from children treated with sodium nitroprusside and it yielded measurable results in 88 of 172 samples (51%), whereas the reference laboratory yielded results in only 19 samples (11%). In all 19 samples, the cobinamide-based method also yielded measurable results. The two methods showed reasonable agreement when analyzed by linear regression, but not when analyzed by a standard error of the estimate or paired t-test. Differences in results between the two methods may be because samples were assayed at different times on different sample types. The cobinamide-based method is applicable to human blood, and can be used in hospital laboratories and emergency rooms. PMID:23653045

  20. Apparatus and method for extraction of chemicals from aquifer remediation effluent water

    DOEpatents

    McMurtrey, Ryan D.; Ginosar, Daniel M.; Moor, Kenneth S.; Shook, G. Michael; Moses, John M.; Barker, Donna L.

    2002-01-01

    An apparatus and method for extraction of chemicals from an aquifer remediation aqueous effluent are provided. The extraction method utilizes a critical fluid for separation and recovery of chemicals employed in remediating aquifers contaminated with hazardous organic substances, and is particularly suited for separation and recovery of organic contaminants and process chemicals used in surfactant-based remediation technologies. The extraction method separates and recovers high-value chemicals from the remediation effluent and minimizes the volume of generated hazardous waste. The recovered chemicals can be recycled to the remediation process or stored for later use.

  1. Method and system for extraction of chemicals from aquifer remediation effluent water

    DOEpatents

    McMurtrey, Ryan D.; Ginosar, Daniel M.; Moor, Kenneth S.; Shook, G. Michael; Barker, Donna L.

    2003-01-01

    A method and system for extraction of chemicals from an groundwater remediation aqueous effluent are provided. The extraction method utilizes a critical fluid for separation and recovery of chemicals employed in remediating groundwater contaminated with hazardous organic substances, and is particularly suited for separation and recovery of organic contaminants and process chemicals used in surfactant-based remediation technologies. The extraction method separates and recovers high-value chemicals from the remediation effluent and minimizes the volume of generated hazardous waste. The recovered chemicals can be recycled to the remediation process or stored for later use.

  2. Extraction methods and food uses of a natural red colorant from dye sorghum.

    PubMed

    Akogou, Folachodé Ug; Kayodé, Ap Polycarpe; den Besten, Heidy Mw; Linnemann, Anita R

    2018-01-01

    The interest in stable natural colorants for food applications continues to grow. A red pigment extracted from the leaf sheaths of a sorghum variety (Sorghum bicolor) with a high content of apigeninidin is widely used as a biocolorant in processed foods in West Africa. This study compared the colour and anthocyanin composition from traditional extraction methods to determine options for improvement and use of the red biocolorant from dye sorghum in the food sector. Sorghum biocolorant was commonly applied in fermented and heated foods. Traditional extraction methods predominantly differed in two aspects, namely the use of an alkaline rock salt (locally known as kanwu) and the temperature of the extraction water. Cool extraction using the alkaline ingredient was more efficient than hot alkaline and hot aqueous extractions in extracting anthocyanins. The apigeninidin content was three times higher in the cool and hot alkaline extracts than in the aqueous extract. Cool and hot alkaline extractions at pH 8-9 were the most efficient methods for extracting apigeninidin from dye sorghum leaf sheaths. Broader use of the sorghum biocolorant in foods requires further research on its effects on nutrient bioavailability and antioxidant activity. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  3. Reference Proteome Extracts for Mass Spec Instrument Performance Validation and Method Development

    PubMed Central

    Rosenblatt, Mike; Urh, Marjeta; Saveliev, Sergei

    2014-01-01

    Biological samples of high complexity are required to test protein mass spec sample preparation procedures and validate mass spec instrument performance. Total cell protein extracts provide the needed sample complexity. However, to be compatible with mass spec applications, such extracts should meet a number of design requirements: compatibility with LC/MS (free of detergents, etc.)high protein integrity (minimal level of protein degradation and non-biological PTMs)compatibility with common sample preparation methods such as proteolysis, PTM enrichment and mass-tag labelingLot-to-lot reproducibility Here we describe total protein extracts from yeast and human cells that meet the above criteria. Two extract formats have been developed: Intact protein extracts with primary use for sample preparation method development and optimizationPre-digested extracts (peptides) with primary use for instrument validation and performance monitoring

  4. Optimization and single-laboratory validation of a method for the determination of flavonolignans in milk thistle seeds by high-performance liquid chromatography with ultraviolet detection.

    PubMed

    Mudge, Elizabeth; Paley, Lori; Schieber, Andreas; Brown, Paula N

    2015-10-01

    Seeds of milk thistle, Silybum marianum (L.) Gaertn., are used for treatment and prevention of liver disorders and were identified as a high priority ingredient requiring a validated analytical method. An AOAC International expert panel reviewed existing methods and made recommendations concerning method optimization prior to validation. A series of extraction and separation studies were undertaken on the selected method for determining flavonolignans from milk thistle seeds and finished products to address the review panel recommendations. Once optimized, a single-laboratory validation study was conducted. The method was assessed for repeatability, accuracy, selectivity, LOD, LOQ, analyte stability, and linearity. Flavonolignan content ranged from 1.40 to 52.86% in raw materials and dry finished products and ranged from 36.16 to 1570.7 μg/mL in liquid tinctures. Repeatability for the individual flavonolignans in raw materials and finished products ranged from 1.03 to 9.88% RSDr, with HorRat values between 0.21 and 1.55. Calibration curves for all flavonolignan concentrations had correlation coefficients of >99.8%. The LODs for the flavonolignans ranged from 0.20 to 0.48 μg/mL at 288 nm. Based on the results of this single-laboratory validation, this method is suitable for the quantitation of the six major flavonolignans in milk thistle raw materials and finished products, as well as multicomponent products containing dandelion, schizandra berry, and artichoke extracts. It is recommended that this method be adopted as First Action Official Method status by AOAC International.

  5. A comparative study: the impact of different lipid extraction methods on current microalgal lipid research

    PubMed Central

    2014-01-01

    Microalgae cells have the potential to rapidly accumulate lipids, such as triacylglycerides that contain fatty acids important for high value fatty acids (e.g., EPA and DHA) and/or biodiesel production. However, lipid extraction methods for microalgae cells are not well established, and there is currently no standard extraction method for the determination of the fatty acid content of microalgae. This has caused a few problems in microlagal biofuel research due to the bias derived from different extraction methods. Therefore, this study used several extraction methods for fatty acid analysis on marine microalga Tetraselmis sp. M8, aiming to assess the potential impact of different extractions on current microalgal lipid research. These methods included classical Bligh & Dyer lipid extraction, two other chemical extractions using different solvents and sonication, direct saponification and supercritical CO2 extraction. Soxhlet-based extraction was used to weigh out the importance of solvent polarity in the algal oil extraction. Coupled with GC/MS, a Thermogravimetric Analyser was used to improve the quantification of microalgal lipid extractions. Among these extractions, significant differences were observed in both, extract yield and fatty acid composition. The supercritical extraction technique stood out most for effective extraction of microalgal lipids, especially for long chain unsaturated fatty acids. The results highlight the necessity for comparative analyses of microalgae fatty acids and careful choice and validation of analytical methodology in microalgal lipid research. PMID:24456581

  6. Comparing Standard and Selective Degradation DNA Extraction Methods: Results from a Field Experiment with Sexual Assault Kits.

    PubMed

    Campbell, Rebecca; Pierce, Steven J; Sharma, Dhruv B; Shaw, Jessica; Feeney, Hannah; Nye, Jeffrey; Schelling, Kristin; Fehler-Cabral, Giannina

    2017-01-01

    A growing number of U.S. cities have large numbers of untested sexual assault kits (SAKs) in police property facilities. Testing older kits and maintaining current case work will be challenging for forensic laboratories, creating a need for more efficient testing methods. We evaluated selective degradation methods for DNA extraction using actual case work from a sample of previously unsubmitted SAKs in Detroit, Michigan. We randomly assigned 350 kits to either standard or selective degradation testing methods and then compared DNA testing rates and CODIS entry rates between the two groups. Continuation-ratio modeling showed no significant differences, indicating that the selective degradation method had no decrement in performance relative to customary methods. Follow-up equivalence tests indicated that CODIS entry rates for the two methods could differ by more than ±5%. Selective degradation methods required less personnel time for testing and scientific review than standard testing. © 2016 American Academy of Forensic Sciences.

  7. A Robust Gradient Based Method for Building Extraction from LiDAR and Photogrammetric Imagery.

    PubMed

    Siddiqui, Fasahat Ullah; Teng, Shyh Wei; Awrangjeb, Mohammad; Lu, Guojun

    2016-07-19

    Existing automatic building extraction methods are not effective in extracting buildings which are small in size and have transparent roofs. The application of large area threshold prohibits detection of small buildings and the use of ground points in generating the building mask prevents detection of transparent buildings. In addition, the existing methods use numerous parameters to extract buildings in complex environments, e.g., hilly area and high vegetation. However, the empirical tuning of large number of parameters reduces the robustness of building extraction methods. This paper proposes a novel Gradient-based Building Extraction (GBE) method to address these limitations. The proposed method transforms the Light Detection And Ranging (LiDAR) height information into intensity image without interpolation of point heights and then analyses the gradient information in the image. Generally, building roof planes have a constant height change along the slope of a roof plane whereas trees have a random height change. With such an analysis, buildings of a greater range of sizes with a transparent or opaque roof can be extracted. In addition, a local colour matching approach is introduced as a post-processing stage to eliminate trees. This stage of our proposed method does not require any manual setting and all parameters are set automatically from the data. The other post processing stages including variance, point density and shadow elimination are also applied to verify the extracted buildings, where comparatively fewer empirically set parameters are used. The performance of the proposed GBE method is evaluated on two benchmark data sets by using the object and pixel based metrics (completeness, correctness and quality). Our experimental results show the effectiveness of the proposed method in eliminating trees, extracting buildings of all sizes, and extracting buildings with and without transparent roof. When compared with current state-of-the-art building

  8. A Robust Gradient Based Method for Building Extraction from LiDAR and Photogrammetric Imagery

    PubMed Central

    Siddiqui, Fasahat Ullah; Teng, Shyh Wei; Awrangjeb, Mohammad; Lu, Guojun

    2016-01-01

    Existing automatic building extraction methods are not effective in extracting buildings which are small in size and have transparent roofs. The application of large area threshold prohibits detection of small buildings and the use of ground points in generating the building mask prevents detection of transparent buildings. In addition, the existing methods use numerous parameters to extract buildings in complex environments, e.g., hilly area and high vegetation. However, the empirical tuning of large number of parameters reduces the robustness of building extraction methods. This paper proposes a novel Gradient-based Building Extraction (GBE) method to address these limitations. The proposed method transforms the Light Detection And Ranging (LiDAR) height information into intensity image without interpolation of point heights and then analyses the gradient information in the image. Generally, building roof planes have a constant height change along the slope of a roof plane whereas trees have a random height change. With such an analysis, buildings of a greater range of sizes with a transparent or opaque roof can be extracted. In addition, a local colour matching approach is introduced as a post-processing stage to eliminate trees. This stage of our proposed method does not require any manual setting and all parameters are set automatically from the data. The other post processing stages including variance, point density and shadow elimination are also applied to verify the extracted buildings, where comparatively fewer empirically set parameters are used. The performance of the proposed GBE method is evaluated on two benchmark data sets by using the object and pixel based metrics (completeness, correctness and quality). Our experimental results show the effectiveness of the proposed method in eliminating trees, extracting buildings of all sizes, and extracting buildings with and without transparent roof. When compared with current state-of-the-art building

  9. Quantitative assessment of tumour extraction from dermoscopy images and evaluation of computer-based extraction methods for an automatic melanoma diagnostic system.

    PubMed

    Iyatomi, Hitoshi; Oka, Hiroshi; Saito, Masataka; Miyake, Ayako; Kimoto, Masayuki; Yamagami, Jun; Kobayashi, Seiichiro; Tanikawa, Akiko; Hagiwara, Masafumi; Ogawa, Koichi; Argenziano, Giuseppe; Soyer, H Peter; Tanaka, Masaru

    2006-04-01

    The aims of this study were to provide a quantitative assessment of the tumour area extracted by dermatologists and to evaluate computer-based methods from dermoscopy images for refining a computer-based melanoma diagnostic system. Dermoscopic images of 188 Clark naevi, 56 Reed naevi and 75 melanomas were examined. Five dermatologists manually drew the border of each lesion with a tablet computer. The inter-observer variability was evaluated and the standard tumour area (STA) for each dermoscopy image was defined. Manual extractions by 10 non-medical individuals and by two computer-based methods were evaluated with STA-based assessment criteria: precision and recall. Our new computer-based method introduced the region-growing approach in order to yield results close to those obtained by dermatologists. The effectiveness of our extraction method with regard to diagnostic accuracy was evaluated. Two linear classifiers were built using the results of conventional and new computer-based tumour area extraction methods. The final diagnostic accuracy was evaluated by drawing the receiver operating curve (ROC) of each classifier, and the area under each ROC was evaluated. The standard deviations of the tumour area extracted by five dermatologists and 10 non-medical individuals were 8.9% and 10.7%, respectively. After assessment of the extraction results by dermatologists, the STA was defined as the area that was selected by more than two dermatologists. Dermatologists selected the melanoma area with statistically smaller divergence than that of Clark naevus or Reed naevus (P = 0.05). By contrast, non-medical individuals did not show this difference. Our new computer-based extraction algorithm showed superior performance (precision, 94.1%; recall, 95.3%) to the conventional thresholding method (precision, 99.5%; recall, 87.6%). These results indicate that our new algorithm extracted a tumour area close to that obtained by dermatologists and, in particular, the border part

  10. Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory; a method supplement for the determination of Fipronil and degradates in water by gas chromatography/mass spectrometry

    USGS Publications Warehouse

    Madsen, James F.; Sandstrom, Mark W.; Zaugg, Steven D.

    2002-01-01

    A method for the isolation and detemrination of fipronil and four of its degradates has been developed. This method adapts an analytical method created by the U.S. Geological Survey National Water Quality Laboratory in 1995 for the determination of a broad range of high-use pesticides typically found in filtered natural-water samples. In 2000, fipronil and four of its degradates were extracted, analyzed, and validated using this method. The recoveries for these five compounds in reagent-water samples fortified at 1 microgram per liter (ug/L) avereraged 98 percent. Initial method detection limits averaged 0.0029 ug/L. The performance of these five new compounds is consistent with the performance of the compounds in the initial method, making it possible to include them in addition to the other 41 pesticides and pesticide degradates in the original method.

  11. Text extraction method for historical Tibetan document images based on block projections

    NASA Astrophysics Data System (ADS)

    Duan, Li-juan; Zhang, Xi-qun; Ma, Long-long; Wu, Jian

    2017-11-01

    Text extraction is an important initial step in digitizing the historical documents. In this paper, we present a text extraction method for historical Tibetan document images based on block projections. The task of text extraction is considered as text area detection and location problem. The images are divided equally into blocks and the blocks are filtered by the information of the categories of connected components and corner point density. By analyzing the filtered blocks' projections, the approximate text areas can be located, and the text regions are extracted. Experiments on the dataset of historical Tibetan documents demonstrate the effectiveness of the proposed method.

  12. Improvement of seawater salt quality by hydro-extraction and re-crystallization methods

    NASA Astrophysics Data System (ADS)

    Sumada, K.; Dewati, R.; Suprihatin

    2018-01-01

    Indonesia is one of the salt producing countries that use sea water as a source of raw materials, the quality of salt produced is influenced by the quality of sea water. The resulting average salt quality contains 85-90% NaCl. The Indonesian National Standard (SNI) for human salt’s consumption sodium chloride content is 94.7 % (dry base) and for industrial salt 98,5 %. In this study developed the re-crystallization without chemical and hydro-extraction method. The objective of this research to choose the best methods based on efficiency. The results showed that re-crystallization method can produce salt with NaCl content 99,21%, while hydro-extraction method content 99,34 % NaCl. The salt produced through both methods can be used as a consumption and industrial salt, Hydro-extraction method is more efficient than re-crystallization method because re-crystallization method requires heat energy.

  13. An adaptive singular spectrum analysis method for extracting brain rhythms of electroencephalography

    PubMed Central

    Hu, Hai; Guo, Shengxin; Liu, Ran

    2017-01-01

    Artifacts removal and rhythms extraction from electroencephalography (EEG) signals are important for portable and wearable EEG recording devices. Incorporating a novel grouping rule, we proposed an adaptive singular spectrum analysis (SSA) method for artifacts removal and rhythms extraction. Based on the EEG signal amplitude, the grouping rule determines adaptively the first one or two SSA reconstructed components as artifacts and removes them. The remaining reconstructed components are then grouped based on their peak frequencies in the Fourier transform to extract the desired rhythms. The grouping rule thus enables SSA to be adaptive to EEG signals containing different levels of artifacts and rhythms. The simulated EEG data based on the Markov Process Amplitude (MPA) EEG model and the experimental EEG data in the eyes-open and eyes-closed states were used to verify the adaptive SSA method. Results showed a better performance in artifacts removal and rhythms extraction, compared with the wavelet decomposition (WDec) and another two recently reported SSA methods. Features of the extracted alpha rhythms using adaptive SSA were calculated to distinguish between the eyes-open and eyes-closed states. Results showed a higher accuracy (95.8%) than those of the WDec method (79.2%) and the infinite impulse response (IIR) filtering method (83.3%). PMID:28674650

  14. A validated solid-liquid extraction method for the HPLC determination of polyphenols in apple tissues Comparison with pressurised liquid extraction.

    PubMed

    Alonso-Salces, Rosa M; Barranco, Alejandro; Corta, Edurne; Berrueta, Luis A; Gallo, Blanca; Vicente, Francisca

    2005-02-15

    A solid-liquid extraction procedure followed by reversed-phase high-performance liquid chromatography (RP-HPLC) coupled with a photodiode array detector (DAD) for the determination of polyphenols in freeze-dried apple peel and pulp is reported. The extraction step consists in sonicating 0.5g of freeze-dried apple tissue with 30mL of methanol-water-acetic acid (30:69:1, v/v/v) containing 2g of ascorbic acid/L, for 10min in an ultrasonic bath. The whole method was validated, concluding that it is a robust method that presents high extraction efficiencies (peel: >91%, pulp: >95%) and appropriate precisions (within day: R.S.D. (n = 5) <5%, and between days: R.S.D. (n = 5) <7%) at the different concentration levels of polyphenols that can be found in apple samples. The method was compared with one previously published, consisting in a pressurized liquid extraction (PLE) followed by RP-HPLC-DAD determination. The advantages and disadvantages of both methods are discussed.

  15. Environmentally friendly microwave-assisted sequential extraction method followed by ICP-OES and ion-chromatographic analysis for rapid determination of sulphur forms in coal samples.

    PubMed

    Mketo, Nomvano; Nomngongo, Philiswa N; Ngila, J Catherine

    2018-05-15

    A rapid three-step sequential extraction method was developed under microwave radiation followed by inductively coupled plasma-optical emission spectroscopic (ICP-OES) and ion-chromatographic (IC) analysis for the determination of sulphur forms in coal samples. The experimental conditions of the proposed microwave-assisted sequential extraction (MW-ASE) procedure were optimized by using multivariate mathematical tools. Pareto charts generated from 2 3 full factorial design showed that, extraction time has insignificant effect on the extraction of sulphur species, therefore, all the sequential extraction steps were performed for 5 min. The optimum values according to the central composite designs and counter plots of the response surface methodology were 200 °C (microwave temperature) and 0.1 g (coal amount) for all the investigated extracting reagents (H 2 O, HCl and HNO 3 ). When the optimum conditions of the proposed MW-ASE procedure were applied in coal CRMs, SARM 18 showed more organic sulphur (72%) and the other two coal CRMs (SARMs 19 and 20) were dominated by sulphide sulphur species (52-58%). The sum of the sulphur forms from the sequential extraction steps have shown consistent agreement (95-96%) with certified total sulphur values on the coal CRM certificates. This correlation, in addition to the good precision (1.7%) achieved by the proposed procedure, suggests that the sequential extraction method is reliable, accurate and reproducible. To safe-guard the destruction of pyritic and organic sulphur forms in extraction step 1, water was used instead of HCl. Additionally, the notorious acidic mixture (HCl/HNO 3 /HF) was replaced by greener reagent (H 2 O 2 ) in the last extraction step. Therefore, the proposed MW-ASE method can be applied in routine laboratories for the determination of sulphur forms in coal and coal related matrices. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. Optimization of Polycyclic Aromatic Hydrocarbon (PAH) Extraction Efficiency Parameters for Sub- and Supercritical Water Extraction (SCWE) Instrument

    NASA Technical Reports Server (NTRS)

    Okada, Asahi A.

    2005-01-01

    Polycyclic aromatic hydrocarbons are a class of molecules composed of multiple, bonded benzene rings. As PAHS are believed to be present on Mars, positive confirmation of their presence on Mars is highly desirable. To extract PAHS, which have low volatility, a fluid extraction method is ideal, and one that does not utilize organic solvents is especially ideal for in situ instrumental analysis. The use of water as a solvent, which at subcritical pressures and temperatures is relatively non-Polar, has significant potential. As SCWE instruments have not yet been commercialized, all instruments are individually-built research prototypes: thus, initial efforts were intended to determine if extraction efficiencies on the JPL-built laboratory-scale SCWE instrument are comparable to differing designs built elsewhere. Samples of soil with certified reference concentrations of PAHs were extracted using SCWE as well as conventional Soxhlet extraction. Continuation of the work would involve extractions on JPL'S newer, portable SCWE instrument prototype to determine its efficiency in extracting PAHs.

  17. DNA decontamination methods for internal quality management in clinical PCR laboratories.

    PubMed

    Wu, Yingping; Wu, Jianyong; Zhang, Zhihui; Cheng, Chen

    2018-03-01

    The polymerase chain reaction (PCR) technique, one of the most commonly applied methods in diagnostic and molecular biology, has a frustrating downside: the occurrence of false-positive signals due to contamination. In previous research, various DNA decontamination methods have been developed to overcome this limitation. Unfortunately, the use of random or poorly focused sampling methods for monitoring air and/or object surfaces leads to the incomplete elimination during decontamination procedures. We herein attempted to develop a novel DNA decontamination method (environmental surveillance, including surface and air sampling) and quality management program for clinical molecular diagnostic laboratories (or clinical PCR laboratories). Here, we performed a step-by-step evaluation of current DNA decontamination methods and developed an effective procedure for assessing the presence of decontaminating DNA via PCR analysis. Performing targeted environmental surveillance by sampling, which reached optimal performance over 2 weeks, and the decontamination process had been verified as reliable. Additionally, the process was validated to not affect PCR amplification efficiency based on a comparative study. In this study, effective guidelines for DNA decontamination were developed. The method employed ensured that surface DNA contamination could be effectively identified and eliminated. Furthermore, our study highlighted the importance of overall quality assurance and good clinical laboratory practices for preventing contamination, which are key factors for compliance with regulatory or accreditation requirements. Taken together, we provided the evidence that the presented scheme ranged from troubleshooting to the elimination of surface contamination, could serve as critical foundation for developing regular environmental surveillance guidelines for PCR laboratories. © 2017 Wiley Periodicals, Inc.

  18. Development of the front end test stand and vessel for extraction and source plasma analyses negative hydrogen ion sources at the Rutherford Appleton Laboratory.

    PubMed

    Lawrie, S R; Faircloth, D C; Letchford, A P; Perkins, M; Whitehead, M O; Wood, T; Gabor, C; Back, J

    2014-02-01

    The ISIS pulsed spallation neutron and muon facility at the Rutherford Appleton Laboratory (RAL) in the UK uses a Penning surface plasma negative hydrogen ion source. Upgrade options for the ISIS accelerator system demand a higher current, lower emittance beam with longer pulse lengths from the injector. The Front End Test Stand is being constructed at RAL to meet the upgrade requirements using a modified ISIS ion source. A new 10% duty cycle 25 kV pulsed extraction power supply has been commissioned and the first meter of 3 MeV radio frequency quadrupole has been delivered. Simultaneously, a Vessel for Extraction and Source Plasma Analyses is under construction in a new laboratory at RAL. The detailed measurements of the plasma and extracted beam characteristics will allow a radical overhaul of the transport optics, potentially yielding a simpler source configuration with greater output and lifetime.

  19. DO TIE LABORATORY BASED ASSESSMENT METHODS REALLY PREDICT FIELD EFFECTS?

    EPA Science Inventory

    Sediment Toxicity Identification and Evaluation (TIE) methods have been developed for both porewaters and whole sediments. These relatively simple laboratory methods are designed to identify specific toxicants or classes of toxicants in sediments; however, the question of whethe...

  20. Comparative study of mobility extraction methods in p-type polycrystalline silicon thin film transistors

    NASA Astrophysics Data System (ADS)

    Liu, Kai; Liu, Yuan; Liu, Yu-Rong; En, Yun-Fei; Li, Bin

    2017-07-01

    Channel mobility in the p-type polycrystalline silicon thin film transistors (poly-Si TFTs) is extracted using Hoffman method, linear region transconductance method and multi-frequency C-V method. Due to the non-negligible errors when neglecting the dependence of gate-source voltage on the effective mobility, the extracted mobility results are overestimated using linear region transconductance method and Hoffman method, especially in the lower gate-source voltage region. By considering of the distribution of localized states in the band-gap, the frequency independent capacitance due to localized charges in the sub-gap states and due to channel free electron charges in the conduction band were extracted using multi-frequency C-V method. Therefore, channel mobility was extracted accurately based on the charge transport theory. In addition, the effect of electrical field dependent mobility degradation was also considered in the higher gate-source voltage region. In the end, the extracted mobility results in the poly-Si TFTs using these three methods are compared and analyzed.

  1. Antifouling booster biocide extraction from marine sediments: a fast and simple method based on vortex-assisted matrix solid-phase extraction.

    PubMed

    Caldas, Sergiane Souza; Soares, Bruno Meira; Abreu, Fiamma; Castro, Ítalo Braga; Fillmann, Gilberto; Primel, Ednei Gilberto

    2018-03-01

    This paper reports the development of an analytical method employing vortex-assisted matrix solid-phase dispersion (MSPD) for the extraction of diuron, Irgarol 1051, TCMTB (2-thiocyanomethylthiobenzothiazole), DCOIT (4,5-dichloro-2-n-octyl-3-(2H)-isothiazolin-3-one), and dichlofluanid from sediment samples. Separation and determination were performed by liquid chromatography tandem-mass spectrometry. Important MSPD parameters, such as sample mass, mass of C18, and type and volume of extraction solvent, were investigated by response surface methodology. Quantitative recoveries were obtained with 2.0 g of sediment sample, 0.25 g of C18 as the solid support, and 10 mL of methanol as the extraction solvent. The MSPD method was suitable for the extraction and determination of antifouling biocides in sediment samples, with recoveries between 61 and 103% and a relative standard deviation lower than 19%. Limits of quantification between 0.5 and 5 ng g -1 were obtained. Vortex-assisted MPSD was shown to be fast and easy to use, with the advantages of low cost and reduced solvent consumption compared to the commonly employed techniques for the extraction of booster biocides from sediment samples. Finally, the developed method was applied to real samples. Results revealed that the developed extraction method is effective and simple, thus allowing the determination of biocides in sediment samples.

  2. Rapid detection of potyviruses from crude plant extracts.

    PubMed

    Silva, Gonçalo; Oyekanmi, Joshua; Nkere, Chukwuemeka K; Bömer, Moritz; Kumar, P Lava; Seal, Susan E

    2018-04-01

    Potyviruses (genus Potyvirus; family Potyviridae) are widely distributed and represent one of the most economically important genera of plant viruses. Therefore, their accurate detection is a key factor in developing efficient control strategies. However, this can sometimes be problematic particularly in plant species containing high amounts of polysaccharides and polyphenols such as yam (Dioscorea spp.). Here, we report the development of a reliable, rapid and cost-effective detection method for the two most important potyviruses infecting yam based on reverse transcription-recombinase polymerase amplification (RT-RPA). The developed method, named 'Direct RT-RPA', detects each target virus directly from plant leaf extracts prepared with a simple and inexpensive extraction method avoiding laborious extraction of high-quality RNA. Direct RT-RPA enables the detection of virus-positive samples in under 30 min at a single low operation temperature (37 °C) without the need for any expensive instrumentation. The Direct RT-RPA tests constitute robust, accurate, sensitive and quick methods for detection of potyviruses from recalcitrant plant species. The minimal sample preparation requirements and the possibility of storing RPA reagents without cold chain storage, allow Direct RT-RPA to be adopted in minimally equipped laboratories and with potential use in plant clinic laboratories and seed certification facilities worldwide. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  3. Evaluation of methods for the quantification of ether extract contents in forage and cattle feces.

    PubMed

    Barbosa, Marcília M; Detmann, Edenio; Valadares, Sebastião C; Detmann, Kelly S C; Franco, Marcia O; Batista, Erick D; Rocha, Gabriel C

    2017-01-01

    The objective of this study was to compare the estimates of ether extract (EE) contents obtained by the Randall method and by the high-temperature method of the American Oil Chemist's Society (AOCS; Am 5-04) in forages (n = 20) and cattle feces (n = 15). The EE contents were quantified by using the Randall extraction or AOCS method and XT4 filter bags or cartridges made of qualitative filter paper (80 g/m²) as containers for the samples. It was also evaluated the loss of particles, and concentration of residual chlorophyll after extraction and the recovery of protein and minerals in the material subjected to extraction. Significant interaction was observed between extraction method and material for EE contents. The EE estimates using the AOCS method were higher, mainly in forages. No loss of particles was observed with different containers. The chlorophyll contents in the residues of cattle feces were not affected by the extraction method; however, residual chlorophyll was lower using the AOCS method in forages. There was complete recovery of the protein and ash after extraction. The results suggest that AOCS method produces higher estimates of EE contents in forages and cattle feces, possibly by providing greater extraction of non-fatty EE.

  4. Deep Learning Methods for Underwater Target Feature Extraction and Recognition

    PubMed Central

    Peng, Yuan; Qiu, Mengran; Shi, Jianfei; Liu, Liangliang

    2018-01-01

    The classification and recognition technology of underwater acoustic signal were always an important research content in the field of underwater acoustic signal processing. Currently, wavelet transform, Hilbert-Huang transform, and Mel frequency cepstral coefficients are used as a method of underwater acoustic signal feature extraction. In this paper, a method for feature extraction and identification of underwater noise data based on CNN and ELM is proposed. An automatic feature extraction method of underwater acoustic signals is proposed using depth convolution network. An underwater target recognition classifier is based on extreme learning machine. Although convolution neural networks can execute both feature extraction and classification, their function mainly relies on a full connection layer, which is trained by gradient descent-based; the generalization ability is limited and suboptimal, so an extreme learning machine (ELM) was used in classification stage. Firstly, CNN learns deep and robust features, followed by the removing of the fully connected layers. Then ELM fed with the CNN features is used as the classifier to conduct an excellent classification. Experiments on the actual data set of civil ships obtained 93.04% recognition rate; compared to the traditional Mel frequency cepstral coefficients and Hilbert-Huang feature, recognition rate greatly improved. PMID:29780407

  5. Current lipid extraction methods are significantly enhanced adding a water treatment step in Chlorella protothecoides.

    PubMed

    Ren, Xiaojie; Zhao, Xinhe; Turcotte, François; Deschênes, Jean-Sébastien; Tremblay, Réjean; Jolicoeur, Mario

    2017-02-11

    Microalgae have the potential to rapidly accumulate lipids of high interest for the food, cosmetics, pharmaceutical and energy (e.g. biodiesel) industries. However, current lipid extraction methods show efficiency limitation and until now, extraction protocols have not been fully optimized for specific lipid compounds. The present study thus presents a novel lipid extraction method, consisting in the addition of a water treatment of biomass between the two-stage solvent extraction steps of current extraction methods. The resulting modified method not only enhances lipid extraction efficiency, but also yields a higher triacylglycerols (TAG) ratio, which is highly desirable for biodiesel production. Modification of four existing methods using acetone, chloroform/methanol (Chl/Met), chloroform/methanol/H 2 O (Chl/Met/H 2 O) and dichloromethane/methanol (Dic/Met) showed respective lipid extraction yield enhancement of 72.3, 35.8, 60.3 and 60.9%. The modified acetone method resulted in the highest extraction yield, with 68.9 ± 0.2% DW total lipids. Extraction of TAG was particularly improved with the water treatment, especially for the Chl/Met/H 2 O and Dic/Met methods. The acetone method with the water treatment led to the highest extraction level of TAG with 73.7 ± 7.3 µg/mg DW, which is 130.8 ± 10.6% higher than the maximum value obtained for the four classical methods (31.9 ± 4.6 µg/mg DW). Interestingly, the water treatment preferentially improved the extraction of intracellular fractions, i.e. TAG, sterols, and free fatty acids, compared to the lipid fractions of the cell membranes, which are constituted of phospholipids (PL), acetone mobile polar lipids and hydrocarbons. Finally, from the 32 fatty acids analyzed for both neutral lipids (NL) and polar lipids (PL) fractions, it is clear that the water treatment greatly improves NL-to-PL ratio for the four standard methods assessed. Water treatment of biomass after the first solvent extraction step

  6. Effects of Extraction Methods on Phytochemicals of Rice Bran Oils Produced from Colored Rice.

    PubMed

    Mingyai, Sukanya; Srikaeo, Khongsak; Kettawan, Aikkarach; Singanusong, Riantong; Nakagawa, Kiyotaka; Kimura, Fumiko; Ito, Junya

    2018-02-01

    Rice bran oil (RBO) especially from colored rice is rich in phytochemicals and has become popular in food, cosmetic, nutraceutical and pharmaceutical applications owing to its offering health benefits. This study determined the contents of phytochemicals including oryzanols, phytosterols, tocopherols (Toc) and tocotrienols (T3) in RBOs extracted using different methods namely cold-press extraction (CPE), solvent extraction (SE) and supercritical CO 2 extraction (SC-CO 2 ). Two colored rice, Red Jasmine rice (RJM, red rice) and Hom-nin rice (HN, black rice), were studied in comparison with the popular Thai fragrant rice Khao Dawk Mali 105 (KDML 105, white rice). RBOs were found to be the rich source of oryzanols, phytosterols, Toc and T3. Rice varieties had a greater effect on the phytochemicals concentrations than extraction methods. HN rice showed the significantly highest concentration of all phytochemicals, followed by RJM and KDML 105 rice, indicating that colored rice contained high concentration of phytochemicals in the oil than non-colored rice. The RBO samples extracted by the CPE method had a greater concentration of the phytochemicals than those extracted by the SC-CO 2 and SE methods, respectively. In terms of phytochemical contents, HN rice extracted using CPE method was found to be the best.

  7. Extraction Method and Analysis of Cannabinoids in Cannabis Olive Oil Preparations.

    PubMed

    Casiraghi, Antonella; Roda, Gabriella; Casagni, Eleonora; Cristina, Cecilia; Musazzi, Umberto Maria; Franzè, Silvia; Rocco, Paolo; Giuliani, Claudia; Fico, Gelsomina; Minghetti, Paola; Gambaro, Veniero

    2018-03-01

    Recently, an increasing number of pharmacists had to supply medicinal products based on Cannabis sativa L. (Cannabaceae), prescribed by physicians to individual patients. Cannabis olive oil preparation is the first choice as a concentrated extract of cannabinoids, even though standardized operative conditions for obtaining it are still not available. In this work, the impact of temperature and extraction time on the concentration of active principles was studied to harmonize the different compounding methods, optimize the extraction process, and reduce the variability among preparations. Moreover, starting from the cannabis inflorescence, the effect of temperature on tetrahydrocannabinolic acid decarboxylation was evaluated. For the analysis, a GC/MS method, as suggested by the Italian Ministry of Health, and a GC/flame ionization detection method were developed, validated, and compared. Georg Thieme Verlag KG Stuttgart · New York.

  8. A Comparison of Protein Extraction Methods Suitable for Gel-Based Proteomic Studies of Aphid Proteins

    PubMed Central

    Cilia, M.; Fish, T.; Yang, X.; Mclaughlin, M.; Thannhauser, T. W.

    2009-01-01

    Protein extraction methods can vary widely in reproducibility and in representation of the total proteome, yet there are limited data comparing protein isolation methods. The methodical comparison of protein isolation methods is the first critical step for proteomic studies. To address this, we compared three methods for isolation, purification, and solubilization of insect proteins. The aphid Schizaphis graminum, an agricultural pest, was the source of insect tissue. Proteins were extracted using TCA in acetone (TCA-acetone), phenol, or multi-detergents in a chaotrope solution. Extracted proteins were solubilized in a multiple chaotrope solution and examined using 1-D and 2-D electrophoresis and compared directly using 2-D Difference Gel Electrophoresis (2-D DIGE). Mass spectrometry was used to identify proteins from each extraction type. We were unable to ascribe the differences in the proteins extracted to particular physical characteristics, cell location, or biological function. The TCA-acetone extraction yielded the greatest amount of protein from aphid tissues. Each extraction method isolated a unique subset of the aphid proteome. The TCA-acetone method was explored further for its quantitative reliability using 2-D DIGE. Principal component analysis showed that little of the variation in the data was a result of technical issues, thus demonstrating that the TCA-acetone extraction is a reliable method for preparing aphid proteins for a quantitative proteomics experiment. These data suggest that although the TCA-acetone method is a suitable method for quantitative aphid proteomics, a combination of extraction approaches is recommended for increasing proteome coverage when using gel-based separation techniques. PMID:19721822

  9. A simple method to extract DNA from hair shafts using enzymatic laundry powder.

    PubMed

    Guan, Zheng; Zhou, Yu; Liu, Jinchuan; Jiang, Xiaoling; Li, Sicong; Yang, Shuming; Chen, Ailiang

    2013-01-01

    A simple method to extract DNA from hair shafts was developed by using enzymatic laundry powder at the first step of the process. The whole extraction can be finished in less than 2 hours. The simple extraction reagent proposed here contains only two cheap components: ordinary enzymatic laundry powder and PCR buffer. After extraction, an ultra sensitive fluorescent nucleic acid stain, PicoGreen, was used for quantifying trace amount of double-stranded DNA in the solution extracted. For further validation of DNA extraction, four primers were employed to amplify DNA microsatellite loci. Both fluorescence spectroscopy and PCR results suggested that this method can extract DNA from hair shafts with good efficiency and repeatability. The study will greatly facilitate the use of hair shafts in future for DNA analyses on genome-wide scale.

  10. Rapid methods for extraction and concentration of poliovirus from oyster tissues.

    PubMed

    Richards, G P; Goldmintz, D; Green, D L; Babinchak, J A

    1982-12-01

    A procedure is discussed for the extraction of poliovirus from oyster meats by modification of several enterovirus extraction techniques. The modified method uses meat extract and Cat-Floc, a polycationic electrolyte, for virus extraction and concentration. Virus recovery from inoculated oyster homogenates is 93-120%. Adsorption of viruses to oyster proteins by acidification of homogenates does not affect virus recovery. Elution of viruses from oyster proteins appears more efficient at pH 9.5 than at pH 8.0. This technique is relatively simple, economical and requires only 2.5 h to complete the combined extraction and concentration procedure.

  11. Laboratory and Self-Report Methods to Assess Reappraisal and Distraction in Youth.

    PubMed

    Bettis, Alexandra H; Henry, Lauren; Prussien, Kemar V; Vreeland, Allison; Smith, Michele; Adery, Laura H; Compas, Bruce E

    2018-06-07

    Coping and emotion regulation are central features of risk and resilience in childhood and adolescence, but research on these constructs has relied on different methods of assessment. The current study aimed to bridge the gap between questionnaire and experimental methods of measuring secondary control coping strategies, specifically distraction and cognitive reappraisal, and examine associations with symptoms of anxiety and depression in youth. A community sample of 70 youth (ages 9-15) completed a novel experimental coping and emotion regulation paradigm and self-report measures of coping and emotion regulation and symptoms. Findings indicate that use of distraction and reappraisal during the laboratory paradigm was associated with lower levels of negative emotion during the task. Youth emotion ratings while implementing distraction, but not reappraisal, during the laboratory task were associated with youth self-reported use of secondary control coping in response to family stress. Youth symptoms of anxiety and depression were also significantly positively associated with negative emotion ratings during the laboratory task, and both laboratory task and self-reported coping and emotion regulation accounted for significant variance in symptoms in youth. Both questionnaire and laboratory methods to assess coping and emotion regulation in youth are important for understanding these processes as possible mechanisms of risk and resilience and continued integration of these methods is a priority for future research.

  12. Effect of extraction method on the yield of furanocoumarins from fruits of Archangelica officinalis Hoffm.

    PubMed

    Waksmundzka-Hajnos, M; Petruczynik, A; Dragan, A; Wianowska, D; Dawidowicz, A L

    2004-01-01

    Optimal conditions for the extraction and analysis of furanocoumarins from fruits of Archangelica officinalis Hoffm. have been determined. The following extraction methods were used: exhaustive extraction in a Soxhlet apparatus, ultrasonication at 25 and 60 degrees C, microwave-assisted solvent extraction in open and closed systems, and accelerated solvent extraction (ASE). In most cases the yields of furanocoumarins were highest using the ASE method. The effects of extracting solvent, temperature and time of extraction using this method were investigated. The highest yield of furanocoumarins by ASE was obtained with methanol at 100-130 degrees C for 10 min. The extraction yields of furanocoumarins from plant material by ultrasonication at 60 degrees C and microwave-assisted solvent extraction in an open system were comparable to the extraction yields obtained in the time- and solvent-consuming exhaustive process involving the Soxhlet apparatus.

  13. [Action of extract and oil neem in the control of Rhipicephalus (Boophilus) microplus (Canestrini, 1887) (Acari: Ixodidae) in laboratory].

    PubMed

    Broglio-Micheletti, Sônia Maria Forti; Dias, Nivia da Silva; Valente, Ellen Carine Neves; de Souza, Leilianne Alves; Lopes, Diego Olympio Peixoto; Dos Santos, Jakeline Maria

    2010-01-01

    Organic plant extracts and emulsified oil of Azadirachta indica A. Juss (Meliaceae) (neem) were studied to evaluate its effects in control of engorged females of Rhipicephalus (Boophilus) microplus (Canestrini, 1887) in the laboratory. Hexane and alcoholic organic extracts, 2% (weight/volume) were used in tests of immersion for 5 minutes, prepared with seeds, solubilized in dimethylsulfoxide (DMSO) to 1%. The experiment was entirely randomized, consisting of 6 treatments and 5 replicates, each represented by 5 ticks. Control groups consisted of untreated females. Based on the results of this work, we can indicate that the seed extract (hexanic fraction) and óleo emulsionável I¹ concentration to 2% have significant adjuvant potential to control the cattle tick, because, cause the mortality in the first days after the treatment and interfere in the reproduction, showing to be an alternative to acaricides normally used.

  14. Accurate facade feature extraction method for buildings from three-dimensional point cloud data considering structural information

    NASA Astrophysics Data System (ADS)

    Wang, Yongzhi; Ma, Yuqing; Zhu, A.-xing; Zhao, Hui; Liao, Lixia

    2018-05-01

    Facade features represent segmentations of building surfaces and can serve as a building framework. Extracting facade features from three-dimensional (3D) point cloud data (3D PCD) is an efficient method for 3D building modeling. By combining the advantages of 3D PCD and two-dimensional optical images, this study describes the creation of a highly accurate building facade feature extraction method from 3D PCD with a focus on structural information. The new extraction method involves three major steps: image feature extraction, exploration of the mapping method between the image features and 3D PCD, and optimization of the initial 3D PCD facade features considering structural information. Results show that the new method can extract the 3D PCD facade features of buildings more accurately and continuously. The new method is validated using a case study. In addition, the effectiveness of the new method is demonstrated by comparing it with the range image-extraction method and the optical image-extraction method in the absence of structural information. The 3D PCD facade features extracted by the new method can be applied in many fields, such as 3D building modeling and building information modeling.

  15. Method of extracting coal from a coal refuse pile

    DOEpatents

    Yavorsky, Paul M.

    1991-01-01

    A method of extracting coal from a coal refuse pile comprises soaking the coal refuse pile with an aqueous alkali solution and distributing an oxygen-containing gas throughout the coal refuse pile for a time period sufficient to effect oxidation of coal contained in the coal refuse pile. The method further comprises leaching the coal refuse pile with an aqueous alkali solution to solubilize and extract the oxidized coal as alkali salts of humic acids and collecting the resulting solution containing the alkali salts of humic acids. Calcium hydroxide may be added to the solution of alkali salts of humic acid to form precipitated humates useable as a low-ash, low-sulfur solid fuel.

  16. EXTRACTION METHODS FOR RECOVERY OF VOLATILE ORGANIC COMPOUNDS FROM FORTIFIED DRY SOILS

    EPA Science Inventory

    Recovery of 8 volatile organic compounds (VOCs) from dry soils, each fortified at 800 ng/g soil, was studied in relation to the extraction method and time of extraction. Extraction procedures studied on desiccator-dried soils were modifications of EPA low-and high-level purge-and...

  17. Viability testing of material derived from Mycobacterium tuberculosis prior to removal from a containment level-III laboratory as part of a Laboratory Risk Assessment Program.

    PubMed

    Blackwood, Kym S; Burdz, Tamara V; Turenne, Christine Y; Sharma, Meenu K; Kabani, Amin M; Wolfe, Joyce N

    2005-01-24

    In the field of clinical mycobacteriology, Mycobacterium tuberculosis (MTB) can be a difficult organism to manipulate due to the restrictive environment of a containment level 3 (CL3) laboratory. Tests for rapid diagnostic work involving smears and molecular methods do not require CL3 practices after the organism has been rendered non-viable. While it has been assumed that after organism deactivation these techniques can be performed outside of a CL3, no conclusive study has consistently confirmed that the organisms are noninfectious after the theoretical 'deactivation' steps. Previous studies have shown that initial steps (such as heating/chemical fixation) may not consistently kill MTB organisms. An inclusive viability study (n = 226) was undertaken to determine at which point handling of culture extraction materials does not necessitate a CL3 environment. Four different laboratory protocols tested for viability included: standard DNA extractions for IS6110 fingerprinting, crude DNA preparations for PCR by boiling and mechanical lysis, protein extractions, and smear preparations. For each protocol, laboratory staff planted a proportion of the resulting material to Bactec 12B medium that was observed for growth for 8 weeks. Of the 208 isolates initially tested, 21 samples grew within the 8-week period. Sixteen (7.7%) of these yielded positive results for MTB that included samples of: deactivated culture resuspensions exposed to 80 degrees C for 20 minutes, smear preparations and protein extractions. Test procedures were consequently modified and tested again (n = 18), resulting in 0% viability. This study demonstrates that it cannot be assumed that conventional practices (i.e. smear preparation) or extraction techniques render the organism non-viable. All methodologies, new and existing, should be examined by individual laboratories to validate the safe removal of material derived from MTB to the outside of a CL3 laboratory. This process is vital to establish in

  18. Metal fractionation in olive oil and urban sewage sludges using the three-stage BCR sequential extraction method and microwave single extractions.

    PubMed

    Pérez Cid, B; Fernández Alborés, A; Fernández Gómez, E; Faliqé López, E

    2001-08-01

    The conventional three-stage BCR sequential extraction method was employed for the fractionation of heavy metals in sewage sludge samples from an urban wastewater treatment plant and from an olive oil factory. The results obtained for Cu, Cr, Ni, Pb and Zn in these samples were compared with those attained by a simplified extraction procedure based on microwave single extractions and using the same reagents as employed in each individual BCR fraction. The microwave operating conditions in the single extractions (heating time and power) were optimized for all the metals studied in order to achieve an extraction efficiency similar to that of the conventional BCR procedure. The measurement of metals in the extracts was carried out by flame atomic absorption spectrometry. The results obtained in the first and third fractions by the proposed procedure were, for all metals, in good agreement with those obtained using the BCR sequential method. Although in the reducible fraction the extraction efficiency of the accelerated procedure was inferior to that of the conventional method, the overall metals leached by both microwave single and sequential extractions were basically the same (recoveries between 90.09 and 103.7%), except for Zn in urban sewage sludges where an extraction efficiency of 87% was achieved. Chemometric analysis showed a good correlation between the results given by the two extraction methodologies compared. The application of the proposed approach to a certified reference material (CRM-601) also provided satisfactory results in the first and third fractions, as it was observed for the sludge samples analysed.

  19. INTER LABORATORY COMBAT HELMET BLUNT IMPACT TEST METHOD COMPARISON

    DTIC Science & Technology

    2018-03-26

    HELMET BLUNT IMPACT TEST METHOD COMPARISON by Tony J. Kayhart Charles A. Hewitt and Jonathan Cyganik March 2018 Final...Report March 2016 – August 2017 Approved for public release; distribution is unlimited U.S. Army Natick Soldier Research ...INTER-LABORATORY COMBAT HELMET BLUNT IMPACT TEST METHOD COMPARISON 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR

  20. Improved Butanol-Methanol (BUME) Method by Replacing Acetic Acid for Lipid Extraction of Biological Samples.

    PubMed

    Cruz, Mutya; Wang, Miao; Frisch-Daiello, Jessica; Han, Xianlin

    2016-07-01

    Extraction of lipids from biological samples is a critical step in lipidomics, especially for shotgun lipidomics where lipid extracts are directly infused into a mass spectrometer. The butanol-methanol (BUME) extraction method was originally developed to extract lipids from plasma samples with 1 % acetic acid. Considering some lipids are sensitive to acidic environments, we modified this protocol by replacing acetic acid with lithium chloride solution and extended the modified extraction to tissue samples. Although no significant reduction of plasmalogen levels in the acidic BUME extracts of rat heart samples was found, the modified method was established to extract various tissue samples, including rat liver, heart, and plasma. Essentially identical profiles of the majority of lipid classes were obtained from the extracts of the modified BUME and traditional Bligh-Dyer methods. However, it was found that neither the original, nor the modified BUME method was suitable for 4-hydroxyalkenal species measurement in biological samples.

  1. Improved Butanol-Methanol (BUME) Method by Replacing Acetic Acid for Lipid Extraction of Biological Samples

    PubMed Central

    Cruz, Mutya; Wang, Miao; Frisch-Daiello, Jessica; Han, Xianlin

    2016-01-01

    Extraction of lipids from biological samples is a critical step in lipidomics, especially for shotgun lipidomics where lipid extracts are directly infused into a mass spectrometer. The butanol-methanol (BUME) extraction method was originally developed to extract lipids from plasma samples with 1% acetic acid. Considering some lipids are sensitive to acidic environments, we modified this protocol by replacing acetic acid with lithium chloride solution and extended the modified extraction to tissue samples. Although no significant reduction of plasmalogen levels in the acidic BUME extracts of rat heart samples was found, the modified method was established to extract various tissue samples, including rat liver, heart, and plasma. Essentially identical profiles of the majority of lipid classes were obtained from the extracts of the modified BUME and traditional Bligh-Dyer methods. However, it was found that neither the original, nor the modified BUME method was suitable for 4-hydroxyalkenal species measurement in biological samples. PMID:27245345

  2. Method for extracting and sequestering carbon dioxide

    DOEpatents

    Rau, Gregory H.; Caldeira, Kenneth G.

    2005-05-10

    A method and apparatus to extract and sequester carbon dioxide (CO.sub.2) from a stream or volume of gas wherein said method and apparatus hydrates CO.sub.2, and reacts the resulting carbonic acid with carbonate. Suitable carbonates include, but are not limited to, carbonates of alkali metals and alkaline earth metals, preferably carbonates of calcium and magnesium. Waste products are metal cations and bicarbonate in solution or dehydrated metal salts, which when disposed of in a large body of water provide an effective way of sequestering CO.sub.2 from a gaseous environment.

  3. Extraction study on uranyl nitrate for energy applications

    NASA Astrophysics Data System (ADS)

    Giri, R.; Nath, G.

    2017-07-01

    Due to the ever-growing demand of energy nuclear reactor materials and the nuclear energy are now considered to be the most critical materials and source of energy for future era. Deposition of nuclear wastes in different industry, nuclear power sector are very much toxic in open environment which are hazardous to living being. There are different methods for extraction and reprocessing of these materials which are cost effective and tedious process. Ultrasonic assisted solvent extraction process is a most efficient and economical way for extraction of such type materials. The presence of third phase in mixing of extractants-diluent pair with aqueous phase imposes the problems in extraction of nuclear reactor materials. The appropriate solvent mixture in proper concentration is an important step in the solvent extraction process. Study of thermo-physical properties helps in selecting an optimum blend for extraction process. In the present work, the extraction of uranium with the binary mixture of Methyl Ethyl Ketone (MEK) and Kerosene was investigated and discussed with the variation of ultrasonic frequency for different temperatures. The result shows that the low frequency and low temperature is suitable environment for extraction. The extraction of uranium by this method is found to be a better result for extraction study in laboratory scale as well as industrial sector.

  4. A Method for Extracting Pigments from Squid Doryteuthis pealeii.

    PubMed

    DiBona, Christopher W; Williams, Thomas L; Dinneen, Sean R; Jones Labadie, Stephanie F; Deravi, Leila F

    2016-11-09

    Cephalopods can undergo rapid and adaptive changes in dermal coloration for sensing, communication, defense, and reproduction purposes. These capabilities are supported in part by the areal expansion and retraction of pigmented organs known as chromatophores. While it is known that the chromatophores contain a tethered network of pigmented granules, their structure-function properties have not been fully detailed. We describe a method for isolating the nanostructured granules in squid Doryteuthis pealeii chromatophores and demonstrate how their associated pigments can be extracted in acidic solvents. To accomplish this, the chromatophore containing dermal layer is first manually isolated using a superficial dissection, and the pigment granules are removed using sonication, centrifugation, and washing cycles. Pigments confined within the purified granules are then extracted via acidic methanol solutions, leaving nanostructures with smaller diameters that are void of visible color. This extraction procedure produces a 58% yield of soluble pigments isolated from granules. Using this method, the composition of the chromatophore pigments can be determined and used to provide insight into the mechanism of adaptive coloration in cephalopods.

  5. Exploring microbial diversity in volcanic environments: a review of methods in DNA extraction.

    PubMed

    Herrera, Aude; Cockell, Charles S

    2007-07-01

    The last decade has been marked by a large number of studies focused on understanding the distribution of microorganisms in volcanic environments. These studies are motivated by the desire to elucidate how the geochemically extreme conditions of such environments can influence microbial diversity both on the surface and in the subsurface of the Earth. The exploration of microbial community diversity has generally not relied on culture-dependent methods, but has been carried out using environmental DNA extraction. Because of the large diversity of chemically and physically complex samples, extracting DNA from volcanic environments is technically challenging. In view of the emerging literature, and our own experience in the optimisation of methods for DNA extraction from volcanic materials, it is timely to provide a methodological comparison. This review highlights and discusses new insights and methods published on DNA extraction methods from volcanic samples, considering the different volcanic environments. A description of a recent method for DNA extraction from basalt and obsidian glass rock samples from Iceland is included. Finally, we discuss these approaches in the wider context of modern work to understand the microbial diversity of volcanic environments.

  6. Method of analysis at the U.S. Geological Survey California Water Science Center, Sacramento Laboratory - determination of haloacetic acid formation potential, method validation, and quality-control practices

    USGS Publications Warehouse

    Zazzi, Barbara C.; Crepeau, Kathryn L.; Fram, Miranda S.; Bergamaschi, Brian A.

    2005-01-01

    An analytical method for the determination of haloacetic acid formation potential of water samples has been developed by the U.S. Geological Survey California Water Science Center Sacramento Laboratory. The haloacetic acid formation potential is measured by dosing water samples with chlorine under specified conditions of pH, temperature, incubation time, darkness, and residual-free chlorine. The haloacetic acids formed are bromochloroacetic acid, bromodichloroacetic acid, dibromochloroacetic acid, dibromoacetic acid, dichloroacetic acid, monobromoacetic acid, monochloroacetic acid, tribromoacetic acid, and trichloroacetic acid. They are extracted, methylated, and then analyzed using a gas chromatograph equipped with an electron capture detector. Method validation experiments were performed to determine the method accuracy, precision, and detection limit for each of the compounds. Method detection limits for these nine haloacetic acids ranged from 0.11 to 0.45 microgram per liter. Quality-control practices include the use of blanks, quality-control samples, calibration verification standards, surrogate recovery, internal standard, matrix spikes, and duplicates.

  7. Conventional laboratory methods for cyanotoxins.

    PubMed

    Lawton, Linda A; Edwards, C

    2008-01-01

    It is clear from the literature that numerous methods are available for most cyanotoxins, although many publications on monitoring data indicate that the favored approach is the use proven, robust methods for individual toxins. The most effective approach is the utilization of a robust rapid screen, where positive samples are followed up by qualitative and quantitative analysis to provide the essential decision making data needed for successful management strategies (Fig. 2). Currently, rapid screens are available for microcystins, saxitoxins and anatoxin-a(s), whilst optimisation and validation is needed, many publications report good correlation with the mouse bioassay and HPLC. There is an urgent need for rapid, simple, and inexpensive assays for cylindrospermopsins, anatoxin-a and BMAA. Although methods exist for analysis of BMAA, the fact that a recent study showed 95% of cyanobacteria producing this, some at levels > 6,000 microg g(-1) dry wt, is of concern and rapid screening followed by robust analysis needed. An ideal approach would be a single method capable of extracting and detecting all cyanotoxins. Several publications describe such approaches using LC-MS, but as expected from a group of compounds with diverse chemistry, there are obvious limitations in recoveries during sample processing, chromatographic performance and sensitivity (Dahlmann et al. 2003, Dell'Aversano et al. 2004, Pietsch et al. 2001). Selection of methods must be based on the application requirements, equipment available and cost. For many organisations it may be more cost effective to out-source the occasional analysis. However, as the incidence of blooms appears to be increasing, the need for more rigorous monitoring is needed, sensible investment is needed to meet recommended guidelines. Most of the methods discussed in this paper are suitable for achieving this goal, although clean-up and concentration is usually necessary for physicochemical methods.

  8. Extraction and determination of arsenic species in leafy vegetables: Method development and application.

    PubMed

    Ma, Li; Yang, Zhaoguang; Kong, Qian; Wang, Lin

    2017-02-15

    Extraction of arsenic (As) species in leafy vegetables was investigated by different combinations of methods and extractants. The extracted As species were separated and determined by HPLC-ICP-MS method. The microwave assisted method using 1% HNO3 as the extractant exhibited satisfactory efficiency (>90%) at 90°C for 1.5h. The proposed method was applied for extracting As species from real leafy vegetables. Thirteen cultivars of leafy vegetables were collected and analyzed. The predominant species in all the investigated vegetable samples were As(III) and As(V). Moreover, both As(III) and As(V) concentrations were positive significant (p<0.01) correlated with total As (tAs) concentration. However, the percentage of As(V) reduced with tAs concentration increasing probably due to the conversion and transformation of As(V) to As(III) after uptake. The hazard quotient results indicated no particular risk to 94.6% of local consumers. Considerably carcinogenic risk by consumption of the leafy vegetables was observed. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. System and method for preparing near-surface heavy oil for extraction using microbial degradation

    DOEpatents

    Busche, Frederick D [Highland Village, TX; Rollins, John B [Southlake, TX; Noyes, Harold J [Golden, CO; Bush, James G [West Richland, WA

    2011-04-12

    A system and method for enhancing the recovery of heavy oil in an oil extraction environment by feeding nutrients to a preferred microbial species (bacteria and/or fungi). A method is described that includes the steps of: sampling and identifying microbial species that reside in the oil extraction environment; collecting fluid property data from the oil extraction environment; collecting nutrient data from the oil extraction environment; identifying a preferred microbial species from the oil extraction environment that can transform the heavy oil into a lighter oil; identifying a nutrient from the oil extraction environment that promotes a proliferation of the preferred microbial species; and introducing the nutrient into the oil extraction environment.

  10. [Extraction method suitable for detection of unheated crustaceans including cephalothorax by ELISA].

    PubMed

    Shibahara, Yusuke; Yamada, Itta; Uesaka, Yoshihiko; Uneo, Noriko; Abe, Akihisa; Ohashi, Eiji; Shiomi, Kazuo

    2009-08-01

    When unheated whole samples of crustaceans (shrimp, prawn and crab) were analyzed with our ELISA kit (FA test EIA-Crustacean 'Nissui') using anti-tropomyosin antibodies, a remarkable reduction in reactivity was recognized. This reduction in activity was found to be due to the digestion of tropomyosin during the extraction process by proteases contained in cephalothorax. To avoid the digestion of tropomyosin by proteases, we developed an extraction method (heating method) suitable for the detection of tropomyosin in unheated crustaceans including cephalothorax. Experiments with unheated whole samples of various species of crustaceans confirmed that the heating method greatly improved the low reactivity in the standard method; the heating method gave extraction efficiencies of as high as 93-107%. Various processed crustaceans with cephalothorax, such as dry products (unheated or weakly heated products) and pickles in soy sauce (unheated products), that showed low reactivity with the standard method were confirmed to give superior results with the heating method. These results indicated that the developed heating method is suitable for detecting unheated crustaceans with cephalothorax by means of the ELISA kit.

  11. Comparison of methods of DNA extraction for real-time PCR in a model of pleural tuberculosis.

    PubMed

    Santos, Ana; Cremades, Rosa; Rodríguez, Juan Carlos; García-Pachón, Eduardo; Ruiz, Montserrat; Royo, Gloria

    2010-01-01

    Molecular methods have been reported to have different sensitivities in the diagnosis of pleural tuberculosis and this may in part be caused by the use of different methods of DNA extraction. Our study compares nine DNA extraction systems in an experimental model of pleural tuberculosis. An inoculum of Mycobacterium tuberculosis was added to 23 pleural liquid samples with different characteristics. DNA was subsequently extracted using nine different methods (seven manual and two automatic) for analysis with real-time PCR. Only two methods were able to detect the presence of M. tuberculosis DNA in all the samples: extraction using columns (Qiagen) and automated extraction with the TNAI system (Roche). The automatic method is more expensive, but requires less time. Almost all the false negatives were because of the difficulty involved in extracting M. tuberculosis DNA, as in general, all the methods studied are capable of eliminating inhibitory substances that block the amplification reaction. The method of M. tuberculosis DNA extraction used affects the results of the diagnosis of pleural tuberculosis by molecular methods. DNA extraction systems that have been shown to be effective in pleural liquid should be used.

  12. 7 CFR 51.1179 - Method of juice extraction.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... STANDARDS) United States Standards for Grades of Florida Oranges and Tangelos Standards for Internal Quality... 7 Agriculture 2 2010-01-01 2010-01-01 false Method of juice extraction. 51.1179 Section 51.1179 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards...

  13. Comparison of Chemical Extraction Methods for Determination of Soil Potassium in Different Soil Types

    NASA Astrophysics Data System (ADS)

    Zebec, V.; Rastija, D.; Lončarić, Z.; Bensa, A.; Popović, B.; Ivezić, V.

    2017-12-01

    Determining potassium supply of soil plays an important role in intensive crop production, since it is the basis for balancing nutrients and issuing fertilizer recommendations for achieving high and stable yields within economic feasibility. The aim of this study was to compare the different extraction methods of soil potassium from arable horizon of different types of soils with ammonium lactate method (KAL), which is frequently used as analytical method for determining the accessibility of nutrients and it is a common method used for issuing fertilizer recommendations in many Europe countries. In addition to the ammonium lactate method (KAL, pH 3.75), potassium was extracted with ammonium acetate (KAA, pH 7), ammonium acetate ethylenediaminetetraacetic acid (KAAEDTA, pH 4.6), Bray (KBRAY, pH 2.6) and with barium chloride (K_{BaCl_2 }, pH 8.1). The analyzed soils were extremely heterogeneous with a wide range of determined values. Soil pH reaction ( {pH_{H_2 O} } ) ranged from 4.77 to 8.75, organic matter content ranged from 1.87 to 4.94% and clay content from 8.03 to 37.07%. In relation to KAL method as the standard method, K_{BaCl_2 } method extracts 12.9% more on average of soil potassium, while in relation to standard method, on average KAA extracts 5.3%, KAAEDTA 10.3%, and KBRAY 27.5% less of potassium. Comparison of analyzed extraction methods of potassium from the soil is of high precision, and most reliable comparison was KAL method with KAAEDTA, followed by a: KAA, K_{BaCl_2 } and KBRAY method. Extremely significant statistical correlation between different extractive methods for determining potassium in the soil indicates that any of the methods can be used to accurately predict the concentration of potassium in the soil, and that carried out research can be used to create prediction model for concentration of potassium based on different methods of extraction.

  14. Efficient method for extracting DNA of parasites causing bovine babesiosis from tick vectors

    USDA-ARS?s Scientific Manuscript database

    The southern cattle tick, Rhipicephalus (Boophilus) microplus, is an economically important pest costing animal agriculture billions of dollars worldwide. This research focuses on a comparison of three different tick DNA extraction methods: phenol-chloroform extraction (method 1), a modified version...

  15. A SIMPLE METHOD FOR THE EXTRACTION AND QUANTIFICATION OF PHOTOPIGMENTS FROM SYMBIODINIUM SPP.

    EPA Science Inventory

    John E. Rogers and Dragoslav Marcovich. Submitted. Simple Method for the Extraction and Quantification of Photopigments from Symbiodinium spp.. Limnol. Oceanogr. Methods. 19 p. (ERL,GB 1192).

    We have developed a simple, mild extraction procedure using methanol which, when...

  16. Critical Evaluation of Soil Pore Water Extraction Methods on a Natural Soil

    NASA Astrophysics Data System (ADS)

    Orlowski, Natalie; Pratt, Dyan; Breuer, Lutz; McDonnell, Jeffrey

    2017-04-01

    Soil pore water extraction is an important component in ecohydrological studies for the measurement of δ2H and δ18O. The effect of pore water extraction technique on resultant isotopic signature is poorly understood. Here we present results of an intercomparison of commonly applied lab-based soil water extraction techniques on a natural soil: high pressure mechanical squeezing, centrifugation, direct vapor equilibration, microwave extraction, and two types of cryogenic extraction systems. We applied these extraction methods to a natural summer-dry (gravimetric water contents ranging from 8% to 15%) glacio-lacustrine, moderately fine textured clayey soil; excavated in 10 cm sampling increments to a depth of 1 meter. Isotope results were analyzed via OA-ICOS and compared for each extraction technique that produced liquid water. From our previous intercomparison study among the same extraction techniques but with standard soils, we discovered that extraction methods are not comparable. We therefore tested the null hypothesis that all extraction techniques would be able to replicate the natural evaporation front in a comparable manner occurring in a summer-dry soil. Our results showed that the extraction technique utilized had a significant effect on the soil water isotopic composition. High pressure mechanical squeezing and vapor equilibration techniques produced similar results with similarly sloped evaporation lines. Due to the nature of soil properties and dryness, centrifugation was unsuccessful in obtaining pore water for isotopic analysis. Cryogenic extraction on both tested techniques produced similar results to each other on a similar sloping evaporation line, but dissimilar with depth.

  17. Novel Approaches to Extraction Methods in Recovery of Capsaicin from Habanero Pepper (CNPH 15.192)

    PubMed Central

    Martins, Frederico S.; Borges, Leonardo L.; Ribeiro, Claudia S. C.; Reifschneider, Francisco J. B.; Conceição, Edemilson C.

    2017-01-01

    Introduction: The objective of this study was to compare three capsaicin extraction methods: Shoxlet, Ultrasound-assisted Extraction (UAE), and Shaker-assisted Extraction (SAE) from Habanero pepper, CNPH 15.192. Materials and Methods: The different parameters evaluated were alcohol degree, time extraction, and solid–solvent ratio using response surface methodology (RSM). Results: The three parameters found significant (p < 0.05) were for UAE and solvent concentration and extraction time for SAE. The optimum conditions for the capsaicin UAE and SAE were similar 95% alcohol degree, 30 minutes and solid–liquid ratio 2 mg/mL. The Soxhlet increased the extraction in 10–25%; however, long extraction times (45 minutes) degraded 2% capsaicin. Conclusion: The extraction of capsaicin was influenced by extraction method and by the operating conditions chosen. The optimized conditions provided savings of time, solvent, and herbal material. Prudent choice of the extraction method is essential to ensure optimal yield of extract, thereby making the study relevant and the knowledge gained useful for further exploitation and application of this resource. SUMMARY Habanero pepper, line CNPH 15.192, possess capsaicin in higher levels when compared with others speciesHigher levels of ethanolic strength are more suitable to obtain a higher levels of capsaicinBox-Behnken design indicates to be useful to explore the best conditions of ultrasound assisted extraction of capsaicin. Abbreviations used: Nomenclature UAE: Ultrasound-assisted Extraction; SAE: Shaker-assisted Extraction. PMID:28808409

  18. Single Laboratory Validated Method for Determination of Microcystins and Nodularin in Ambient Freshwaters by Solid Phase Extraction and Liquid Chromatography/ Tandem Mass Spectrometry (LC/MS/MS)

    EPA Pesticide Factsheets

    This document is a standardized, single laboratory validated liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the detection of cyanotoxins—microsystins and nodularin (combined intracellular and extracellular)—in ambient freshwaters.

  19. Method of simultaneous stir bar sorptive extraction of phenethylamines and THC metabolite from urine.

    PubMed

    Goto, Yoshiyuki; Takeda, Shiho; Araki, Toshinori; Fuchigami, Takayuki

    2011-10-01

    Stir bar sorptive extraction is a technique used for extracting target substances from various aqueous matrixes such as environmental water, food, and biological samples. This type of extraction is carried out by rotating a coated stir bar is rotated in the sample solution. In particular, Twister bar is a commercial stir bar that is coated with polydimethylsiloxane (PDMS) and used to perform sorptive extraction. In this study, we developed a method for simultaneous detection of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, and a Δ(9)-tetrahydrocannabiniol (THC) metabolite in human urine. For extracting the target analytes, the Twister bar was simply stirred in the sample in the presence of a derivatizing agent. Using this technique, phenethylamines and the acidic THC metabolite can be simultaneously extracted from human urine. This method also enables the extraction of trace amounts of these substances with good reproducibility and high selectivity. The proposed method offers many advantages over other extraction-based approaches and is therefore well suited for screening psychoactive substances in urine specimens.

  20. Development and Single-Laboratory Validation of a Liquid Chromatography Tandem Mass Spectrometry Method for Quantitation of Tetrodotoxin in Mussels and Oysters.

    PubMed

    Turner, Andrew D; Boundy, Michael J; Rapkova, Monika Dhanji

    2017-09-01

    In recent years, evidence has grown for the presence of tetrodotoxin (TTX) in bivalve mollusks, leading to the potential for consumers of contaminated products to be affected by Tetrodotoxin Shellfish Poisoning (TSP). A single-laboratory validation was conducted for the hydrophilic interaction LC (HILIC) tandem MS (MS/MS) analysis of TTX in common mussels and Pacific oysters-the bivalve species that have been found to contain TTXs in the United Kingdom in recent years. The method consists of a single-step dispersive extraction in 1% acetic acid, followed by a carbon SPE cleanup step before dilution and instrumental analysis. The full method was developed as a rapid tool for the quantitation of TTX, as well as for the associated analogs 4-epi-TTX; 5,6,11-trideoxy TTX; 11-nor TTX-6-ol; 5-deoxy TTX; and 4,9-anhydro TTX. The method can also be run as the acquisition of TTX together with paralytic shellfish toxins. Results demonstrated acceptable method performance characteristics for specificity, linearity, recovery, ruggedness, repeatability, matrix variability, and within-laboratory reproducibility for the analysis of TTX. The LOD and LOQ were fit-for-purpose in comparison to the current action limit for TTX enforced in The Netherlands. In addition, aspects of method performance (LOD, LOQ, and within-laboratory reproducibility) were found to be satisfactory for three other TTX analogs (11-nor TTX-6-ol, 5-deoxy TTX, and 4,9-anhydro TTX). The method was found to be practical and suitable for use in regulatory testing, providing rapid turnaround of sample analysis. Plans currently underway on a full collaborative study to validate a HILIC-MS/MS method for paralytic shellfish poisoning toxins will be extended to include TTX in order to generate international acceptance, ultimately for use as an alternative official control testing method should regulatory controls be adopted.

  1. A proposed method for enhanced eigen-pair extraction using finite element methods: Theory and application

    NASA Technical Reports Server (NTRS)

    Jara-Almonte, J.; Mitchell, L. D.

    1988-01-01

    The paper covers two distinct parts: theory and application. The goal of this work was the reduction of model size with an increase in eigenvalue/vector accuracy. This method is ideal for the condensation of large truss- or beam-type structures. The theoretical approach involves the conversion of a continuum transfer matrix beam element into an 'Exact' dynamic stiffness element. This formulation is implemented in a finite element environment. This results in the need to solve a transcendental eigenvalue problem. Once the eigenvalue is determined the eigenvectors can be reconstructed with any desired spatial precision. No discretization limitations are imposed on the reconstruction. The results of such a combined finite element and transfer matrix formulation is a much smaller FEM eigenvalue problem. This formulation has the ability to extract higher eigenvalues as easily and as accurately as lower eigenvalues. Moreover, one can extract many more eigenvalues/vectors from the model than the number of degrees of freedom in the FEM formulation. Typically, the number of eigenvalues accurately extractable via the 'Exact' element method are at least 8 times the number of degrees of freedom. In contrast, the FEM usually extracts one accurate (within 5 percent) eigenvalue for each 3-4 degrees of freedom. The 'Exact' element results in a 20-30 improvement in the number of accurately extractable eigenvalues and eigenvectors.

  2. Rice hull smoke extract inactivates Salmonella Typhimurium in laboratory media and protects infected mice against mortality

    USDA-ARS?s Scientific Manuscript database

    A recently discovered and characterized rice hull liquid smoke extract was tested for bactericidal activity against Salmonella Typhimurium using the disc-agar method. The Minimum Inhibitory Concentration (MIC) value of rice hull smoke extract was found to be 0.822% (v/v). The in vivo antibacterial a...

  3. Highly Effective DNA Extraction Method for Nuclear Short Tandem Repeat Testing of Skeletal Remains from Mass Graves

    PubMed Central

    Davoren, Jon; Vanek, Daniel; Konjhodzić, Rijad; Crews, John; Huffine, Edwin; Parsons, Thomas J.

    2007-01-01

    Aim To quantitatively compare a silica extraction method with a commonly used phenol/chloroform extraction method for DNA analysis of specimens exhumed from mass graves. Methods DNA was extracted from twenty randomly chosen femur samples, using the International Commission on Missing Persons (ICMP) silica method, based on Qiagen Blood Maxi Kit, and compared with the DNA extracted by the standard phenol/chloroform-based method. The efficacy of extraction methods was compared by real time polymerase chain reaction (PCR) to measure DNA quantity and the presence of inhibitors and by amplification with the PowerPlex 16 (PP16) multiplex nuclear short tandem repeat (STR) kit. Results DNA quantification results showed that the silica-based method extracted on average 1.94 ng of DNA per gram of bone (range 0.25-9.58 ng/g), compared with only 0.68 ng/g by the organic method extracted (range 0.0016-4.4880 ng/g). Inhibition tests showed that there were on average significantly lower levels of PCR inhibitors in DNA isolated by the organic method. When amplified with PP16, all samples extracted by silica-based method produced 16 full loci profiles, while only 75% of the DNA extracts obtained by organic technique amplified 16 loci profiles. Conclusions The silica-based extraction method showed better results in nuclear STR typing from degraded bone samples than a commonly used phenol/chloroform method. PMID:17696302

  4. METHOD 544. DETERMINATION OF MICROCYSTINS AND ...

    EPA Pesticide Factsheets

    Method 544 is an accurate and precise analytical method to determine six microcystins (including MC-LR) and nodularin in drinking water using solid phase extraction and liquid chromatography tandem mass spectrometry (SPE-LC/MS/MS). The advantage of this SPE-LC/MS/MS is its sensitivity and ability to speciate the microcystins. This method development task establishes sample preservation techniques, sample concentration and analytical procedures, aqueous and extract holding time criteria and quality control procedures. Draft Method 544 undergone a multi-laboratory verification to ensure other laboratories can implement the method and achieve the quality control measures specified in the method. It is anticipated that Method 544 may be used in UCMR 4 to collect nationwide occurrence data for selected microcystins in drinking water. The purpose of this research project is to develop an accurate and precise analytical method to concentrate and determine selected MCs and nodularin in drinking water.

  5. Comparison of methods for extracting kafirin proteins from sorghum distillers dried grains with solubles.

    PubMed

    Wang, Ying; Tilley, Michael; Bean, Scott; Sun, X Susan; Wang, Donghai

    2009-09-23

    Use of coproducts generated during fermentation is important to the overall economics of biofuel production. The main coproduct from grain-based ethanol production is distillers dried grains with solubles (DDGS). High in protein, DDGS is a potential source of protein for many bioindustrial applications such as adhesives and resins. The objective of this research was to characterize the composition as well as chemical and physical properties of kafirin proteins from sorghum DDGS with various extraction methods including use of acetic acid, HCl-ethanol and NaOH-ethanol under reducing conditions. Extraction conditions affected purity and thermal properties of the extracted kafirin proteins. Extraction yields of 44.2, 24.2, and 56.8% were achieved by using acetic acid, HCl-ethanol and NaOH-ethanol, respectively. Acetic acid and NaOH-ethanol produced protein with higher purity than kafirins extracted with the HCl-ethanol protocol. The acetic acid extraction protocol produced protein with the highest purity, 98.9%. Several techniques were used to evaluate structural, molecular and thermal properties of kairin extracts. FTIR showed alpha-helix dominated in all three samples, with only a small portion of beta-sheet present. Electrophoresis results showed alpha(1), alpha(2) band and beta kafirins were present in all three extracts. Glass transition peaks of the extracts were shown by DSC to be approximately 230 degrees C. Kafirin degraded at 270-290 degrees C. Size exclusion chromatography revealed that the acetic acid and HCl-ethanol based extraction methods tended to extract more high molecular weight protein than the NaOH-ethanol based method. Reversed phase high-performance liquid chromatography showed that the gamma kafirins were found only in extracts from the NaOH-ethanol extraction method.

  6. Comparison of manual methods of extracting genomic DNA from dried blood spots collected on different cards: implications for clinical practice.

    PubMed

    Molteni, C G; Terranova, L; Zampiero, A; Galeone, C; Principi, N; Esposito, S

    2013-01-01

    Isolating genomic DNA from blood samples is essential when studying the associations between genetic variants and susceptibility to a given clinical condition, or its severity. This study of three extraction techniques and two types of commercially available cards involved 219 children attending our outpatient pediatric clinic for follow-up laboratory tests after they had been hospitalised. An aliquot of venous blood was drawn into plastic tubes without additives and, after several inversions, 80 microL were put on circles of common paper cards and Whatman FTA-treated cards. Three extraction methods were compared: the Qiagen Investigator, Gensolve, and Masterpure. The best method in terms of final DNA yield was Masterpure, which led to a significantly higher yield regardless of the type of card (p less than 0.001), followed by Qiagen Investigator and Gensolve. Masterpure was also the best in terms of price, seemed to be simple and reliable, and required less hands-on time than other techniques. These conclusions support the use of Masterpure in studies that evaluate the associations between genetic variants and the severity or prevalence of infectious diseases.

  7. Development of Solvent Extraction Approach to Recycle Enriched Molybdenum Material

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

    Tkac, Peter; Brown, M. Alex; Sen, Sujat

    2016-06-01

    Argonne National Laboratory, in cooperation with Oak Ridge National Laboratory and NorthStar Medical Technologies, LLC, is developing a recycling process for a solution containing valuable Mo-100 or Mo-98 enriched material. Previously, Argonne had developed a recycle process using a precipitation technique. However, this process is labor intensive and can lead to production of large volumes of highly corrosive waste. This report discusses an alternative process to recover enriched Mo in the form of ammonium heptamolybdate by using solvent extraction. Small-scale experiments determined the optimal conditions for effective extraction of high Mo concentrations. Methods were developed for removal of ammonium chloridemore » from the molybdenum product of the solvent extraction process. In large-scale experiments, very good purification from potassium and other elements was observed with very high recovery yields (~98%).« less

  8. Evaluation Of A Powder-Free DNA Extraction Method For Skeletal Remains.

    PubMed

    Harrel, Michelle; Mayes, Carrie; Gangitano, David; Hughes-Stamm, Sheree

    2018-02-07

    Bones are often recovered in forensic investigations, including missing persons and mass disasters. While traditional DNA extraction methods rely on grinding bone into powder prior to DNA purification, the TBone Ex buffer (DNA Chip Research Inc.) digests bone chips without powdering. In this study, six bones were extracted using the TBone Ex kit in conjunction with the PrepFiler ® BTA™ DNA extraction kit (Thermo Fisher Scientific) both manually and via an automated platform. Comparable amounts of DNA were recovered from a 50 mg bone chip using the TBone Ex kit and 50 mg of powdered bone with the PrepFiler ® BTA™ kit. However, automated DNA purification decreased DNA yield (p < 0.05). Nevertheless, short tandem repeat (STR) success was comparable across all methods tested. This study demonstrates that digestion of whole bone fragments is an efficient alternative to powdering bones for DNA extraction without compromising downstream STR profile quality. © 2018 American Academy of Forensic Sciences.

  9. A MISO-ARX-Based Method for Single-Trial Evoked Potential Extraction.

    PubMed

    Yu, Nannan; Wu, Lingling; Zou, Dexuan; Chen, Ying; Lu, Hanbing

    2017-01-01

    In this paper, we propose a novel method for solving the single-trial evoked potential (EP) estimation problem. In this method, the single-trial EP is considered as a complex containing many components, which may originate from different functional brain sites; these components can be distinguished according to their respective latencies and amplitudes and are extracted simultaneously by multiple-input single-output autoregressive modeling with exogenous input (MISO-ARX). The extraction process is performed in three stages: first, we use a reference EP as a template and decompose it into a set of components, which serve as subtemplates for the remaining steps. Then, a dictionary is constructed with these subtemplates, and EPs are preliminarily extracted by sparse coding in order to roughly estimate the latency of each component. Finally, the single-trial measurement is parametrically modeled by MISO-ARX while characterizing spontaneous electroencephalographic activity as an autoregression model driven by white noise and with each component of the EP modeled by autoregressive-moving-average filtering of the subtemplates. Once optimized, all components of the EP can be extracted. Compared with ARX, our method has greater tracking capabilities of specific components of the EP complex as each component is modeled individually in MISO-ARX. We provide exhaustive experimental results to show the effectiveness and feasibility of our method.

  10. a Landmark Extraction Method Associated with Geometric Features and Location Distribution

    NASA Astrophysics Data System (ADS)

    Zhang, W.; Li, J.; Wang, Y.; Xiao, Y.; Liu, P.; Zhang, S.

    2018-04-01

    Landmark plays an important role in spatial cognition and spatial knowledge organization. Significance measuring model is the main method of landmark extraction. It is difficult to take account of the spatial distribution pattern of landmarks because that the significance of landmark is built in one-dimensional space. In this paper, we start with the geometric features of the ground object, an extraction method based on the target height, target gap and field of view is proposed. According to the influence region of Voronoi Diagram, the description of target gap is established to the geometric representation of the distribution of adjacent targets. Then, segmentation process of the visual domain of Voronoi K order adjacent is given to set up target view under the multi view; finally, through three kinds of weighted geometric features, the landmarks are identified. Comparative experiments show that this method has a certain coincidence degree with the results of traditional significance measuring model, which verifies the effectiveness and reliability of the method and reduces the complexity of landmark extraction process without losing the reference value of landmark.

  11. Laboratory Evaluations of the Fractions Efficacy of Annona senegalensis (Annonaceae) Leaf Extract on Immature Stage Development of Malarial and Filarial Mosquito Vectors

    PubMed Central

    Lame, Younoussa; Nukenine, Elias Nchiwan; Pierre, Danga Yinyang Simon; Elijah, Ajaegbu Eze; Esimone, Charles Okechukwu

    2015-01-01

    Background: Within the framework to control mosquitoes, ovicidal, larvicidal and pupicidal activity of Annona senegalensis leaf extract and its 4 fractions against Anopheles gambiae and Culex quinquefasciatus were evaluated in the laboratory conditions. Methods: Ovicidal test was performed by submitting at least 100 eggs of mosquitoes to 125, 250, 500, 1000 and 2000 ppm concentrations, while larvicidal and pupicidal effects were assessed by submitting 25 larvae or pupae to the concentrations of 2500, 1250, 625 and 312.5 ppm of plant extract or fractions of A. senegalensis. Results: The eggs of An. gambiae were most affected by N-hexane (0.00% hatchability) and chloroform (03.67% hatchability) fractions compared to Cx. quinquefasciatus where at least 25 % hatchability were recorded at 2000 ppm. For larvicidal test, N-hexane (LC50= 298.8 ppm) and chloroform (LC50= 418.3 ppm) fractions were more effective than other fractions on An. gambiae larvae while, a moderate effectiveness was also observed with N-hexane (LC50= 2087.6 ppm), chloroform (LC50= 9010.1 ppm) fractions on Cx. quinquefasciatus larvae. The highest mortality percent of the pupae were also recorded with N-hexane and chloroform fractions on An. gambiae at 2500 ppm. As for Cx. quinquefasciatus only 50 % and 36 % mortality were recorded with N-hexane and chloroform fractions respectively. Conclusion: The extract of A. senegalensis was toxic on immature stage of mosquito species tested. By splitting methanolic crude extract, only N-hexane and chloroform fractions were revealed to possess a mosquitocidal effects and could be considered and utilized for future immature mosquito vectors control. PMID:26623434

  12. Planetary Gears Feature Extraction and Fault Diagnosis Method Based on VMD and CNN.

    PubMed

    Liu, Chang; Cheng, Gang; Chen, Xihui; Pang, Yusong

    2018-05-11

    Given local weak feature information, a novel feature extraction and fault diagnosis method for planetary gears based on variational mode decomposition (VMD), singular value decomposition (SVD), and convolutional neural network (CNN) is proposed. VMD was used to decompose the original vibration signal to mode components. The mode matrix was partitioned into a number of submatrices and local feature information contained in each submatrix was extracted as a singular value vector using SVD. The singular value vector matrix corresponding to the current fault state was constructed according to the location of each submatrix. Finally, by training a CNN using singular value vector matrices as inputs, planetary gear fault state identification and classification was achieved. The experimental results confirm that the proposed method can successfully extract local weak feature information and accurately identify different faults. The singular value vector matrices of different fault states have a distinct difference in element size and waveform. The VMD-based partition extraction method is better than ensemble empirical mode decomposition (EEMD), resulting in a higher CNN total recognition rate of 100% with fewer training times (14 times). Further analysis demonstrated that the method can also be applied to the degradation recognition of planetary gears. Thus, the proposed method is an effective feature extraction and fault diagnosis technique for planetary gears.

  13. Qualitative and quantitative evaluation of the genomic DNA extracted from GMO and non-GMO foodstuffs with four different extraction methods.

    PubMed

    Peano, Clelia; Samson, Maria Cristina; Palmieri, Luisa; Gulli, Mariolina; Marmiroli, Nelson

    2004-11-17

    The presence of DNA in foodstuffs derived from or containing genetically modified organisms (GMO) is the basic requirement for labeling of GMO foods in Council Directive 2001/18/CE (Off. J. Eur. Communities 2001, L1 06/2). In this work, four different methods for DNA extraction were evaluated and compared. To rank the different methods, the quality and quantity of DNA extracted from standards, containing known percentages of GMO material and from different food products, were considered. The food products analyzed derived from both soybean and maize and were chosen on the basis of the mechanical, technological, and chemical treatment they had been subjected to during processing. Degree of DNA degradation at various stages of food production was evaluated through the amplification of different DNA fragments belonging to the endogenous genes of both maize and soybean. Genomic DNA was extracted from Roundup Ready soybean and maize MON810 standard flours, according to four different methods, and quantified by real-time Polymerase Chain Reaction (PCR), with the aim of determining the influence of the extraction methods on the DNA quantification through real-time PCR.

  14. Inter-laboratory variation in the chemical analysis of acidic forest soil reference samples from eastern North America

    USGS Publications Warehouse

    Ross, Donald S.; Bailiey, Scott W; Briggs, Russell D; Curry, Johanna; Fernandez, Ivan J.; Fredriksen, Guinevere; Goodale, Christine L.; Hazlett, Paul W.; Heine, Paul R; Johnson, Chris E.; Larson, John T; Lawrence, Gregory B.; Kolka, Randy K; Ouimet, Rock; Pare, D; Richter, Daniel D.; Shirmer, Charles D; Warby, Richard A.F.

    2015-01-01

    Long-term forest soil monitoring and research often requires a comparison of laboratory data generated at different times and in different laboratories. Quantifying the uncertainty associated with these analyses is necessary to assess temporal changes in soil properties. Forest soil chemical properties, and methods to measure these properties, often differ from agronomic and horticultural soils. Soil proficiency programs do not generally include forest soil samples that are highly acidic, high in extractable Al, low in extractable Ca and often high in carbon. To determine the uncertainty associated with specific analytical methods for forest soils, we collected and distributed samples from two soil horizons (Oa and Bs) to 15 laboratories in the eastern United States and Canada. Soil properties measured included total organic carbon and nitrogen, pH and exchangeable cations. Overall, results were consistent despite some differences in methodology. We calculated the median absolute deviation (MAD) for each measurement and considered the acceptable range to be the median 6 2.5 3 MAD. Variability among laboratories was usually as low as the typical variability within a laboratory. A few areas of concern include a lack of consistency in the measurement and expression of results on a dry weight basis, relatively high variability in the C/N ratio in the Bs horizon, challenges associated with determining exchangeable cations at concentrations near the lower reporting range of some laboratories and the operationally defined nature of aluminum extractability. Recommendations include a continuation of reference forest soil exchange programs to quantify the uncertainty associated with these analyses in conjunction with ongoing efforts to review and standardize laboratory methods.

  15. An improved facile method for extraction and determination of steroidal saponins in Tribulus terrestris by focused microwave-assisted extraction coupled with GC-MS.

    PubMed

    Li, Tianlin; Zhang, Zhuomin; Zhang, Lan; Huang, Xinjian; Lin, Junwei; Chen, Guonan

    2009-12-01

    An improved fast method for extraction of steroidal saponins in Tribulus terrestris based on the use of focus microwave-assisted extraction (FMAE) is proposed. Under optimized conditions, four steroidal saponins were extracted from Tribulus terrestris and identified by GC-MS, which are Tigogenin (TG), Gitogenin (GG), Hecogenin (HG) and Neohecogenin (NG). One of the most important steroidal saponins, namely TG was quantified finally. The recovery of TG was in the range of 86.7-91.9% with RSD<5.2%. The convention heating reflux extraction was also conducted in order to validate the reliability of this new FMAE method. The yield of total steroidal saponins was 90.3% in a one-step FMAE, while the yield of 65.0% was achieved during heating reflux extraction, and the extraction time was reduced from 3 h to 5 min by using less solvent. The method was successfully applied to analyze the steroidal saponins of Tribulus terrestris from different areas of occurrence. The difference in chromatographic characteristics of steroidal saponins was proved to be related to the different areas of occurrence. The results showed that FMAE-GC-MS is a simple, rapid, solvent-saving method for the extraction and determination of steroidal saponins in Tribulus terrestris.

  16. New Methods May Reduce Needs, Can't Replace Laboratory Animals

    ERIC Educational Resources Information Center

    Leeper, E. M.

    1976-01-01

    Discusses the symposium between the scientific community and the animal welfare community in which the consensus was absolute: new laboratory methods can complement but not replace intact animals. (LS)

  17. Antioxidant, antinociceptive and anti-inflammatory properties of the aqueous and ethanolic leaf extracts of Andrographis paniculata in some laboratory animals.

    PubMed

    Adedapo, Adeolu Alex; Adeoye, Bisi Olajumoke; Sofidiya, Margaret Oluwatoyin; Oyagbemi, Ademola Adetokunbo

    2015-07-01

    The study was designed to evaluate the anti-inflammatory, analgesic and antioxidant properties of Andrographis paniculata leaf extracts in laboratory animals. The dried and powdered leaves of the plant were subjected to phytochemical and proximate analyses. Its mineral content was also determined. Acute toxicity experiments were first performed to determine a safe dose level. The plant material was extracted using water and ethanol as solvents. These extracts were then used to test for the anti-inflammatory, analgesic and antioxidant properties of the plant. The anti-inflammatory tests included carrageenan-induced and histamine-induced paw oedema. The analgesic tests conducted were formalin paw lick test and acetic acid writhing test. The antioxidant activities of the extracts of A. paniculata were determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), total polyphenol (TP) and 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) using ascorbic acid as standard for both DPPH and FRAP, and gallic acid as a standard for both TP and ABTS. The acute toxicity experiment demonstrated that the plant is safe at high doses even at 1600 mg/kg. It was observed that the ethanolic extract of A. paniculata had higher antioxidant activity than the aqueous extract. The experiments using both extracts may suggest that the extracts of A. paniculata leaves possess anti-inflammatory, analgesic and antioxidant properties, although the ethanolic extract seemed to have higher biological properties than the aqueous extract. The results from this study may have justified the plant's folkloric use for medicinal purpose.

  18. The study on the extraction method of indirubin from Indigo Naturalis.

    PubMed

    Cheng, Li-hui

    2007-01-01

    To discuss how to extract the indirubin of the higher purity from Indigo Naturalis. After extracting the indirubin from Indigo Naturalis by acetic ether, compare the separation effect of the extract in Si gel column with that in alumina column. The indirubin in Indigo Naturalis by Si gel column chromatography is chosen. The purity of indirubin is 98.3% after Si gel column chromatography. The indiruhbin of the higher purity was attained by the simple experimental method.

  19. Intercomparison of Lab-Based Soil Water Extraction Methods for Stable Water Isotope Analysis

    NASA Astrophysics Data System (ADS)

    Pratt, D.; Orlowski, N.; McDonnell, J.

    2016-12-01

    The effect of pore water extraction technique on resultant isotopic signature is poorly understood. Here we present results of an intercomparison of five common lab-based soil water extraction techniques: high pressure mechanical squeezing, centrifugation, direct vapor equilibration, microwave extraction, and cryogenic extraction. We applied five extraction methods to two physicochemically different standard soil types (silty sand and clayey loam) that were oven-dried and rewetted with water of known isotopic composition at three different gravimetric water contents (8, 20, and 30%). We tested the null hypothisis that all extraction techniques would provide the same isotopic result independent from soil type and water content. Our results showed that the extraction technique had a significant effect on the soil water isotopic composition. Each method exhibited deviations from spiked reference water, with soil type and water content showing a secondary effect. Cryogenic extraction showed the largest deviations from the reference water, whereas mechanical squeezing and centrifugation provided the closest match to the reference water for both soil types. We also compared results for each extraction technique that produced liquid water on both an OA-ICOS and IRMS; differences between them were negligible.

  20. a Probability-Based Statistical Method to Extract Water Body of TM Images with Missing Information

    NASA Astrophysics Data System (ADS)

    Lian, Shizhong; Chen, Jiangping; Luo, Minghai

    2016-06-01

    Water information cannot be accurately extracted using TM images because true information is lost in some images because of blocking clouds and missing data stripes, thereby water information cannot be accurately extracted. Water is continuously distributed in natural conditions; thus, this paper proposed a new method of water body extraction based on probability statistics to improve the accuracy of water information extraction of TM images with missing information. Different disturbing information of clouds and missing data stripes are simulated. Water information is extracted using global histogram matching, local histogram matching, and the probability-based statistical method in the simulated images. Experiments show that smaller Areal Error and higher Boundary Recall can be obtained using this method compared with the conventional methods.

  1. Comparative evaluation of different extraction and quantification methods for forensic RNA analysis.

    PubMed

    Grabmüller, Melanie; Madea, Burkhard; Courts, Cornelius

    2015-05-01

    Since about 2005, there is increasing interest in forensic RNA analysis whose versatility may very favorably complement traditional DNA profiling in forensic casework. There is, however, no method available specifically dedicated for extraction of RNA from forensically relevant sample material. In this study we compared five commercially available and commonly used RNA extraction kits and methods (mirVana™ miRNA Isolation Kit Ambion; Trizol® Reagent, Invitrogen; NucleoSpin® miRNA Kit Macherey-Nagel; AllPrep DNA/RNA Mini Kit and RNeasy® Mini Kit both Qiagen) to assess their relative effectiveness of yielding RNA of good quality and their compatibility with co-extraction of DNA amenable to STR profiling. We set up samples of small amounts of dried blood, liquid saliva, semen and buccal mucosa that were aged for different time intervals for co-extraction of RNA and DNA. RNA quality was assessed by determination of 'RNA integrity number' (RIN) and quantitative PCR based expression analysis. DNA quality was assessed via monitoring STR typing success rates. By comparison, the different methods exhibited considerable differences between RNA and DNA yields, RNA quality values and expression levels, and STR profiling success, with the AllPrep DNA/RNA Mini Kit and the NucleoSpin® miRNA Kit excelling at DNA co-extraction and RNA results, respectively. Overall, there was no 'best' method to satisfy all demands of comprehensible co-analysis of RNA and DNA and it appears that each method has specific merits and flaws. We recommend to cautiously choose from available methods and align its characteristics with the needs of the experimental setting at hand. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  2. Development and validation of an extraction method for the analysis of perfluoroalkyl substances in human hair.

    PubMed

    Kim, Da-Hye; Oh, Jeong-Eun

    2017-05-01

    Human hair has many advantages as a non-invasive sample; however, analytical methods for detecting perfluoroalkyl substances (PFASs) in human hair are still in the development stage. Therefore, the aim of this study was to develop and validate a method for monitoring 11 PFASs in human hair. Solid-phase extraction (SPE), ion-pairing extraction (IPE), a combined method (SPE+IPE) and solvent extraction with ENVI-carb clean-up were compared to develop an optimal extraction method using two types of hair sample (powder and piece forms). Analysis of PFASs was performed using liquid chromatography and tandem mass spectrometry. Among the four different extraction procedures, the SPE method using powdered hair showed the best extraction efficiency and recoveries ranged from 85.8 to 102%. The method detection limits for the SPE method were 0.114-0.796 ng/g and good precision (below 10%) and accuracy (66.4-110%) were obtained. In light of these results, SPE is considered the optimal method for PFAS extraction from hair. It was also successfully used to detect PFASs in human hair samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. [Comparison on extraction of volatile oils from Lithospermum erythrorhizon by different methods].

    PubMed

    Yang, Ri-fu; Huang, Ping-ping; Qiu, Tai-qiu; Fan, Xiao-dan

    2011-02-01

    To extract the volatile oils from Lithospermum erythrorhizon via ultrasound-enhanced sub-critical water extraction (USWE) and compare with ultrasound-enhanced solvent extraction (USE) and steam distillation extraction (SD). The extraction yield of the volatile oils, the containing components of extract, the effect of scanvenging activities on free radical DPPH and reducing activities as well as the inhibitory on escherichia coli and staphylococcus aureus were investigated. The extraction yield of volatile oils by USWE, USE and SD were 2.39%, 1.93% and 0.62%, respectively, the extracts by three methods all contained six major components, but the extracts by SD and USE contained more impurities. The inhibitory effect on escherichia coli and staphylococcus aureus of the extract by SD and its reducing action were the best,but those by USWE were the worst. the extraction yield of volatile oils by USWE is the highest, and it contains less impurities based on the worst in reducing power and inhibitory effects.

  4. Specificity of laboratory trail following by the argentine ant,Iridomyrmex humilis (Mayr), to (Z)-9-hexadecenal, analogs, and gaster extract.

    PubMed

    Van Vorhis Key, S E; Baker, T C

    1982-07-01

    In laboratory trail-following bioassays of Argentine ant workers,Iridomyrmex humilis (Mayr), the geometric isomer, (E)-9-hexadecenal, of the trail pheromone component (Z)-9-hexadecenal elicited insignificant trail following as did the potentially more stable formate analogs, (Z)-7-tetradecenyl formate, (E)-7-tetradecenyl formate, and tetradecyl formate. Further, in direct choice tests, workers showed no preference for gaster extract trails (0.002 ant equiv/cm) over trails of (Z)-9-hexadecenal (0.2 ng/cm). Moreover, a 10-fold increase in synthetic trail concentration to 2.0 ng/cm caused (Z)-9-hexadecenal trails to be significantly preferred over gaster extract trails by trail-following ants.

  5. Comparison of manual and automated nucleic acid extraction methods from clinical specimens for microbial diagnosis purposes.

    PubMed

    Wozniak, Aniela; Geoffroy, Enrique; Miranda, Carolina; Castillo, Claudia; Sanhueza, Francia; García, Patricia

    2016-11-01

    The choice of nucleic acids (NAs) extraction method for molecular diagnosis in microbiology is of major importance because of the low microbial load, different nature of microorganisms, and clinical specimens. The NA yield of different extraction methods has been mostly studied using spiked samples. However, information from real human clinical specimens is scarce. The purpose of this study was to compare the performance of a manual low-cost extraction method (Qiagen kit or salting-out extraction method) with the automated high-cost MagNAPure Compact method. According to cycle threshold values for different pathogens, MagNAPure is as efficient as Qiagen for NA extraction from noncomplex clinical specimens (nasopharyngeal swab, skin swab, plasma, respiratory specimens). In contrast, according to cycle threshold values for RNAseP, MagNAPure method may not be an appropriate method for NA extraction from blood. We believe that MagNAPure versatility reduced risk of cross-contamination and reduced hands-on time compensates its high cost. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Hot Salsa: A Laboratory Exercise Exploring the Scientific Method.

    ERIC Educational Resources Information Center

    Levri, Edward P.; Levri, Maureen A.

    2003-01-01

    Presents a laboratory exercise on spicy food and body temperature that introduces the scientific method to introductory biology students. Suggests that when students perform their own experiments which they have developed, it helps with their understanding of and confidence in doing science. (Author/SOE)

  7. Chemical composition and antibacterial activity of Cordia verbenacea extracts obtained by different methods.

    PubMed

    Michielin, Eliane M Z; Salvador, Ana A; Riehl, Carlos A S; Smânia, Artur; Smânia, Elza F A; Ferreira, Sandra R S

    2009-12-01

    The present study describes the chemical composition and the antibacterial activity of extracts from Cordia verbenacea DC (Borraginaceae), a traditional medicinal plant that grows widely along the southeastern coast of Brazil. The extracts were obtained using different extraction techniques: high-pressure operations and low-pressure methods. The high-pressure technique was applied to obtain C. verbenacea extracts using pure CO(2) and CO(2) with co-solvent at pressures up to 30MPa and temperatures of 30, 40 and 50 degrees C. Organic solvents such as n-hexane, ethyl acetate, ethanol, acetone and dichloromethane were used to obtain extracts by low-pressure processes. The antibacterial activity of the extracts was also subjected to screening against four strains of bacteria using the agar dilution method. The extraction yields were up to 5.0% w/w and up to 8.6% w/w for supercritical fluid extraction with pure CO(2) and with ethyl acetate as co-solvent, respectively, while the low-pressure extraction indicates yields up to 24.0% w/w in the soxhlet extraction using water and aqueous mixture with 50% ethanol as solvents. The inhibitory activity of the extracts in gram-positive bacteria was significantly higher than in gram-negative. The quantification and the identification of the extracts recovered were accomplished using GC/MS analysis. The most important components identified in the extract were artemetin, beta-sitosterol, alpha-humulene and beta-caryophyllene, among others.

  8. Application of solid/liquid extraction for the gravimetric determination of lipids in royal jelly.

    PubMed

    Antinelli, Jean-François; Davico, Renée; Rognone, Catherine; Faucon, Jean-Paul; Lizzani-Cuvelier, Louisette

    2002-04-10

    Gravimetric lipid determination is a major parameter for the characterization and the authentication of royal jelly quality. A solid/liquid extraction was compared to the reference method, which is based on liquid/liquid extraction. The amount of royal jelly and the time of the extraction were optimized in comparison to the reference method. Boiling/rinsing ratio and spread of royal jelly onto the extraction thimble were identified as critical parameters, resulting in good accuracy and precision for the alternative method. Comparison of reproducibility and repeatability of both methods associated with gas chromatographic analysis of the composition of the extracted lipids showed no differences between the two methods. As the intra-laboratory validation tests were comparable to the reference method, while offering rapidity and a decrease in amount of solvent used, it was concluded that the proposed method should be used with no modification of quality criteria and norms established for royal jelly characterization.

  9. An efficient and scalable extraction and quantification method for algal derived biofuel.

    PubMed

    Lohman, Egan J; Gardner, Robert D; Halverson, Luke; Macur, Richard E; Peyton, Brent M; Gerlach, Robin

    2013-09-01

    Microalgae are capable of synthesizing a multitude of compounds including biofuel precursors and other high value products such as omega-3-fatty acids. However, accurate analysis of the specific compounds produced by microalgae is important since slight variations in saturation and carbon chain length can affect the quality, and thus the value, of the end product. We present a method that allows for fast and reliable extraction of lipids and similar compounds from a range of algae, followed by their characterization using gas chromatographic analysis with a focus on biodiesel-relevant compounds. This method determines which range of biologically synthesized compounds is likely responsible for each fatty acid methyl ester (FAME) produced; information that is fundamental for identifying preferred microalgae candidates as a biodiesel source. Traditional methods of analyzing these precursor molecules are time intensive and prone to high degrees of variation between species and experimental conditions. Here we detail a new method which uses microwave energy as a reliable, single-step cell disruption technique to extract lipids from live cultures of microalgae. After extractable lipid characterization (including lipid type (free fatty acids, mono-, di- or tri-acylglycerides) and carbon chain length determination) by GC-FID, the same lipid extracts are transesterified into FAMEs and directly compared to total biodiesel potential by GC-MS. This approach provides insight into the fraction of total FAMEs derived from extractable lipids compared to FAMEs derived from the residual fraction (i.e. membrane bound phospholipids, sterols, etc.). This approach can also indicate which extractable lipid compound, based on chain length and relative abundance, is responsible for each FAME. This method was tested on three species of microalgae; the marine diatom Phaeodactylum tricornutum, the model Chlorophyte Chlamydomonas reinhardtii, and the freshwater green alga Chlorella vulgaris

  10. Novel Approaches to Extraction Methods in Recovery of Capsaicin from Habanero Pepper (CNPH 15.192).

    PubMed

    Martins, Frederico S; Borges, Leonardo L; Ribeiro, Claudia S C; Reifschneider, Francisco J B; Conceição, Edemilson C

    2017-07-01

    The objective of this study was to compare three capsaicin extraction methods: Shoxlet, Ultrasound-assisted Extraction (UAE), and Shaker-assisted Extraction (SAE) from Habanero pepper, CNPH 15.192. The different parameters evaluated were alcohol degree, time extraction, and solid-solvent ratio using response surface methodology (RSM). The three parameters found significant ( p < 0.05) were for UAE and solvent concentration and extraction time for SAE. The optimum conditions for the capsaicin UAE and SAE were similar 95% alcohol degree, 30 minutes and solid-liquid ratio 2 mg/mL. The Soxhlet increased the extraction in 10-25%; however, long extraction times (45 minutes) degraded 2% capsaicin. The extraction of capsaicin was influenced by extraction method and by the operating conditions chosen. The optimized conditions provided savings of time, solvent, and herbal material. Prudent choice of the extraction method is essential to ensure optimal yield of extract, thereby making the study relevant and the knowledge gained useful for further exploitation and application of this resource. Habanero pepper , line CNPH 15.192, possess capsaicin in higher levels when compared with others speciesHigher levels of ethanolic strength are more suitable to obtain a higher levels of capsaicinBox-Behnken design indicates to be useful to explore the best conditions of ultrasound assisted extraction of capsaicin. Abbreviations used: Nomenclature UAE: Ultrasound-assisted Extraction; SAE: Shaker-assisted Extraction.

  11. Laboratory Sequence in Computational Methods for Introductory Chemistry

    NASA Astrophysics Data System (ADS)

    Cody, Jason A.; Wiser, Dawn C.

    2003-07-01

    A four-exercise laboratory sequence for introductory chemistry integrating hands-on, student-centered experience with computer modeling has been designed and implemented. The progression builds from exploration of molecular shapes to intermolecular forces and the impact of those forces on chemical separations made with gas chromatography and distillation. The sequence ends with an exploration of molecular orbitals. The students use the computers as a tool; they build the molecules, submit the calculations, and interpret the results. Because of the construction of the sequence and its placement spanning the semester break, good laboratory notebook practices are reinforced and the continuity of course content and methods between semesters is emphasized. The inclusion of these techniques in the first year of chemistry has had a positive impact on student perceptions and student learning.

  12. Enhancing to method for extracting Social network by the relation existence

    NASA Astrophysics Data System (ADS)

    Elfida, Maria; Matyuso Nasution, M. K.; Sitompul, O. S.

    2018-01-01

    To get the trusty information about the social network extracted from the Web requires a reliable method, but for optimal resultant required the method that can overcome the complexity of information resources. This paper intends to reveal ways to overcome the constraints of social network extraction leading to high complexity by identifying relationships among social actors. By changing the treatment of the procedure used, we obtain the complexity is smaller than the previous procedure. This has also been demonstrated in an experiment by using the denial sample.

  13. An Efficient Extraction Method for Fragrant Volatiles from Jasminum sambac (L.) Ait.

    PubMed

    Ye, Qiuping; Jin, Xinyi; Zhu, Xinliang; Lin, Tongxiang; Hao, Zhilong; Yang, Qian

    2015-01-01

    The sweet smell of aroma of Jasminum sambac (L.) Ait. is releasing while the flowers are blooming. Although components of volatile oil have been extensively studied, there are problematic issues, such as low efficiency of yield, flavour distortion. Here, the subcritical fluid extraction (SFE) was performed to extract fragrant volatiles from activated carbon that had absorbed the aroma of jasmine flowers. This novel method could effectively obtain main aromatic compounds with quality significantly better than solvent extraction (SE). Based on the analysis data with response surface methodology (RSM), we optimized the extraction conditions which consisted of a temperature of 44°C, a solvent-to-material ratio of 3.5:1, and an extraction time of 53 min. Under these conditions, the extraction yield was 4.91%. Furthermore, the key jasmine essence oil components, benzyl acetate and linalool, increase 7 fold and 2 fold respectively which lead to strong typical smell of the jasmine oil. The new method can reduce spicy components which lead to the essential oils smelling sweeter. Thus, the quality of the jasmine essence oil was dramatically improved and yields based on the key component increased dramatically. Our results provide a new effective technique for extracting fragrant volatiles from jasmine flowers.

  14. Alternative method of removing otoliths from sturgeon

    USGS Publications Warehouse

    Chalupnicki, Marc A.; Dittman, Dawn E.

    2016-01-01

    Extracting the otoliths (ear bones) from fish that have very thick skulls can be difficult and very time consuming. The common practice of making a transverse vertical incision on the top of the skull with a hand or electrical saw may damage the otolith if not performed correctly. Sturgeons (Acipenseridae) are one family in particular that have a very large and thick skull. A new laboratory method entering the brain cavity from the ventral side of the fish to expose the otoliths was easier than other otolith extraction methods found in the literature. Methods reviewed in the literature are designed for the field and are more efficient at processing large quantities of fish quickly. However, this new technique was designed to be more suited for a laboratory setting when time is not pressing and successful extraction from each specimen is critical. The success of finding and removing otoliths using this technique is very high and does not compromise the structure in any manner. This alternative technique is applicable to other similar fish species for extracting the otoliths.

  15. Methods for pore water extraction from unsaturated zone tuff, Yucca Mountain, Nevada

    USGS Publications Warehouse

    Scofield, K.M.

    2006-01-01

    Assessing the performance of the proposed high-level radioactive waste repository at Yucca Mountain, Nevada, requires an understanding of the chemistry of the water that moves through the host rock. The uniaxial compression method used to extract pore water from samples of tuffaceous borehole core was successful only for nonwelded tuff. An ultracentrifugation method was adopted to extract pore water from samples of the densely welded tuff of the proposed repository horizon. Tests were performed using both methods to determine the efficiency of pore water extraction and the potential effects on pore water chemistry. Test results indicate that uniaxial compression is most efficient for extracting pore water from nonwelded tuff, while ultracentrifugation is more successful in extracting pore water from densely welded tuff. Pore water splits collected from a single nonwelded tuff core during uniaxial compression tests have shown changes in pore water chemistry with increasing pressure for calcium, chloride, sulfate, and nitrate. Pore water samples collected from the intermediate pressure ranges should prevent the influence of re-dissolved, evaporative salts and the addition of ion-deficient water from clays and zeolites. Chemistry of pore water splits from welded and nonwelded tuffs using ultracentrifugation indicates that there is no substantial fractionation of solutes.

  16. Swab Protocol for Rapid Laboratory Diagnosis of Cutaneous Anthrax

    PubMed Central

    Marston, Chung K.; Bhullar, Vinod; Baker, Daniel; Rahman, Mahmudur; Hossain, M. Jahangir; Chakraborty, Apurba; Khan, Salah Uddin; Hoffmaster, Alex R.

    2012-01-01

    The clinical laboratory diagnosis of cutaneous anthrax is generally established by conventional microbiological methods, such as culture and directly straining smears of clinical specimens. However, these methods rely on recovery of viable Bacillus anthracis cells from swabs of cutaneous lesions and often yield negative results. This study developed a rapid protocol for detection of B. anthracis on clinical swabs. Three types of swabs, flocked-nylon, rayon, and polyester, were evaluated by 3 extraction methods, the swab extraction tube system (SETS), sonication, and vortex. Swabs were spiked with virulent B. anthracis cells, and the methods were compared for their efficiency over time by culture and real-time PCR. Viability testing indicated that the SETS yielded greater recovery of B. anthracis from 1-day-old swabs; however, reduced viability was consistent for the 3 extraction methods after 7 days and nonviability was consistent by 28 days. Real-time PCR analysis showed that the PCR amplification was not impacted by time for any swab extraction method and that the SETS method provided the lowest limit of detection. When evaluated using lesion swabs from cutaneous anthrax outbreaks, the SETS yielded culture-negative, PCR-positive results. This study demonstrated that swab extraction methods differ in their efficiency of recovery of viable B. anthracis cells. Furthermore, the results indicated that culture is not reliable for isolation of B. anthracis from swabs at ≥7 days. Thus, we recommend the use of the SETS method with subsequent testing by culture and real-time PCR for diagnosis of cutaneous anthrax from clinical swabs of cutaneous lesions. PMID:23035192

  17. Evaluation of extraction methods for ochratoxin A detection in cocoa beans employing HPLC.

    PubMed

    Mishra, Rupesh K; Catanante, Gaëlle; Hayat, Akhtar; Marty, Jean-Louis

    2016-01-01

    Cocoa is an important ingredient for the chocolate industry and for many food products. However, it is prone to contamination by ochratoxin A (OTA), which is highly toxic and potentially carcinogenic to humans. In this work, four different extraction methods were tested and compared based on their recoveries. The best protocol was established which involves an organic solvent-free extraction method for the detection of OTA in cocoa beans using 1% sodium hydrogen carbonate (NaHCO3) in water within 30 min. The extraction method is rapid (as compared with existing methods), simple, reliable and practical to perform without complex experimental set-ups. The cocoa samples were freshly extracted and cleaned-up using immunoaffinity column (IAC) for HPLC analysis using a fluorescence detector. Under the optimised condition, the limit of detection (LOD) and limit of quantification (LOQ) for OTA were 0.62 and 1.25 ng ml(-1) respectively in standard solutions. The method could successfully quantify OTA in naturally contaminated samples. Moreover, good recoveries of OTA were obtained up to 86.5% in artificially spiked cocoa samples, with a maximum relative standard deviation (RSD) of 2.7%. The proposed extraction method could determine OTA at the level 1.5 µg kg(-)(1), which surpassed the standards set by the European Union for cocoa (2 µg kg(-1)). In addition, an efficiency comparison of IAC and molecular imprinted polymer (MIP) column was also performed and evaluated.

  18. Validation of a One-Step Method for Extracting Fatty Acids from Salmon, Chicken and Beef Samples.

    PubMed

    Zhang, Zhichao; Richardson, Christine E; Hennebelle, Marie; Taha, Ameer Y

    2017-10-01

    Fatty acid extraction methods are time-consuming and expensive because they involve multiple steps and copious amounts of extraction solvents. In an effort to streamline the fatty acid extraction process, this study compared the standard Folch lipid extraction method to a one-step method involving a column that selectively elutes the lipid phase. The methods were tested on raw beef, salmon, and chicken. Compared to the standard Folch method, the one-step extraction process generally yielded statistically insignificant differences in chicken and salmon fatty acid concentrations, percent composition and weight percent. Initial testing showed that beef stearic, oleic and total fatty acid concentrations were significantly lower by 9-11% with the one-step method as compared to the Folch method, but retesting on a different batch of samples showed a significant 4-8% increase in several omega-3 and omega-6 fatty acid concentrations with the one-step method relative to the Folch. Overall, the findings reflect the utility of a one-step extraction method for routine and rapid monitoring of fatty acids in chicken and salmon. Inconsistencies in beef concentrations, although minor (within 11%), may be due to matrix effects. A one-step fatty acid extraction method has broad applications for rapidly and routinely monitoring fatty acids in the food supply and formulating controlled dietary interventions. © 2017 Institute of Food Technologists®.

  19. Study of Burn Scar Extraction Automatically Based on Level Set Method using Remote Sensing Data

    PubMed Central

    Liu, Yang; Dai, Qin; Liu, JianBo; Liu, ShiBin; Yang, Jin

    2014-01-01

    Burn scar extraction using remote sensing data is an efficient way to precisely evaluate burn area and measure vegetation recovery. Traditional burn scar extraction methodologies have no well effect on burn scar image with blurred and irregular edges. To address these issues, this paper proposes an automatic method to extract burn scar based on Level Set Method (LSM). This method utilizes the advantages of the different features in remote sensing images, as well as considers the practical needs of extracting the burn scar rapidly and automatically. This approach integrates Change Vector Analysis (CVA), Normalized Difference Vegetation Index (NDVI) and the Normalized Burn Ratio (NBR) to obtain difference image and modifies conventional Level Set Method Chan-Vese (C-V) model with a new initial curve which results from a binary image applying K-means method on fitting errors of two near-infrared band images. Landsat 5 TM and Landsat 8 OLI data sets are used to validate the proposed method. Comparison with conventional C-V model, OSTU algorithm, Fuzzy C-mean (FCM) algorithm are made to show that the proposed approach can extract the outline curve of fire burn scar effectively and exactly. The method has higher extraction accuracy and less algorithm complexity than that of the conventional C-V model. PMID:24503563

  20. Planetary Gears Feature Extraction and Fault Diagnosis Method Based on VMD and CNN

    PubMed Central

    Cheng, Gang; Chen, Xihui

    2018-01-01

    Given local weak feature information, a novel feature extraction and fault diagnosis method for planetary gears based on variational mode decomposition (VMD), singular value decomposition (SVD), and convolutional neural network (CNN) is proposed. VMD was used to decompose the original vibration signal to mode components. The mode matrix was partitioned into a number of submatrices and local feature information contained in each submatrix was extracted as a singular value vector using SVD. The singular value vector matrix corresponding to the current fault state was constructed according to the location of each submatrix. Finally, by training a CNN using singular value vector matrices as inputs, planetary gear fault state identification and classification was achieved. The experimental results confirm that the proposed method can successfully extract local weak feature information and accurately identify different faults. The singular value vector matrices of different fault states have a distinct difference in element size and waveform. The VMD-based partition extraction method is better than ensemble empirical mode decomposition (EEMD), resulting in a higher CNN total recognition rate of 100% with fewer training times (14 times). Further analysis demonstrated that the method can also be applied to the degradation recognition of planetary gears. Thus, the proposed method is an effective feature extraction and fault diagnosis technique for planetary gears. PMID:29751671

  1. Use of an enzyme-assisted method to improve protein extraction from olive leaves.

    PubMed

    Vergara-Barberán, M; Lerma-García, M J; Herrero-Martínez, J M; Simó-Alfonso, E F

    2015-02-15

    The improvement of protein extraction from olive leaves using an enzyme-assisted protocol has been investigated. Using a cellulase enzyme (Celluclast® 1.5L), different parameters that affect the extraction process, such as the influence and amount of organic solvent, enzyme amount, pH and extraction temperature and time, were optimised. The influence of these factors was examined using the standard Bradford assay and the extracted proteins were characterised by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum extraction parameters were: 30% acetonitrile, 5% (v/v) Celluclast® 1.5L at pH 5.0 and 55°C for 15min. Under these conditions, several protein extracts from olive leaves of different genetic variety (with a total protein amount comprised between 1.87 and 6.64mgg(-1)) were analysed and compared by SDS-PAGE, showing differences in their electrophoretic protein profiles. The developed enzyme-assisted extraction method has shown a faster extraction, higher recovery and reduced solvent usage with respect to the use of the non-enzymatic methods described in literature. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. A novel optimised and validated method for analysis of multi-residues of pesticides in fruits and vegetables by microwave-assisted extraction (MAE)-dispersive solid-phase extraction (d-SPE)-retention time locked (RTL)-gas chromatography-mass spectrometry with Deconvolution reporting software (DRS).

    PubMed

    Satpathy, Gouri; Tyagi, Yogesh Kumar; Gupta, Rajinder Kumar

    2011-08-01

    A rapid, effective and ecofriendly method for sensitive screening and quantification of 72 pesticides residue in fruits and vegetables, by microwave-assisted extraction (MAE) followed by dispersive solid-phase extraction (d-SPE), retention time locked (RTL) capillary gas-chromatographic separation in trace ion mode mass spectrometric determination has been validated as per ISO/IEC: 17025:2005. Identification and reporting with total and extracted ion chromatograms were facilitated to a great extent by Deconvolution reporting software (DRS). For all compounds LOD were 0.002-0.02mg/kg and LOQ were 0.025-0.100mg/kg. Correlation coefficients of the calibration curves in the range of 0.025-0.50mg/kg were >0.993. To validate matrix effects repeatability, reproducibility, recovery and overall uncertainty were calculated for the 35 matrices at 0.025, 0.050 and 0.100mg/kg. Recovery ranged between 72% and 114% with RSD of <20% for repeatability and intermediate precision. The reproducibility of the method was evaluated by an inter laboratory participation and Z score obtained within ±2. Copyright © 2011 Elsevier Ltd. All rights reserved.

  3. A Generic multi-dimensional feature extraction method using multiobjective genetic programming.

    PubMed

    Zhang, Yang; Rockett, Peter I

    2009-01-01

    In this paper, we present a generic feature extraction method for pattern classification using multiobjective genetic programming. This not only evolves the (near-)optimal set of mappings from a pattern space to a multi-dimensional decision space, but also simultaneously optimizes the dimensionality of that decision space. The presented framework evolves vector-to-vector feature extractors that maximize class separability. We demonstrate the efficacy of our approach by making statistically-founded comparisons with a wide variety of established classifier paradigms over a range of datasets and find that for most of the pairwise comparisons, our evolutionary method delivers statistically smaller misclassification errors. At very worst, our method displays no statistical difference in a few pairwise comparisons with established classifier/dataset combinations; crucially, none of the misclassification results produced by our method is worse than any comparator classifier. Although principally focused on feature extraction, feature selection is also performed as an implicit side effect; we show that both feature extraction and selection are important to the success of our technique. The presented method has the practical consequence of obviating the need to exhaustively evaluate a large family of conventional classifiers when faced with a new pattern recognition problem in order to attain a good classification accuracy.

  4. Effects of Different Extraction Methods on the Extraction Rates of Five Chemical Ingredients of Swertia mussotii Franch by UPLC-ESI-MS/MS

    NASA Astrophysics Data System (ADS)

    Xiong, Yaokun; Zhou, Lifen; Zhao, Yonghong; Liu, Yun; Liu, Xia; Zhu, Genhua; Yan, Zhihong; Liu, Zhiyong

    2018-01-01

    Objective: To compare the effects of three extraction methods (ultrasound, reflux and percolation) on the contents of gentiopicroside, mangiferin, swertiamain, sweroside and oleanolic acid in Swertia mussotii Franch. Method: The contents of five components were determined by UPLC-ESI/MS. In the solvent system, eluent A was 0.05% (v: v) formic acid with 1mM/L ammonium acetate aqueous solution and eluent B was acetonitrile. Chromatographic separations were achieved using an Agilent EC-C18 column (4.6×100 mm, 2.7 um) at 30 °C. The flow rate was set at 0.8mL/min. The compound ionization was adopted at negative ionization mode by electro spray ionization (ESI). The quantification was performed in multiple reaction monitoring (MRM). Results: The linear ranges of swertiamarin, gentiopicroside, mangiferin, sweroside and oleanolic acid are 80∼7450 ng/mL, 103∼6600 ng/mL, 100∼8000 ng/mL, 130∼8450 ng/mL, 100∼7000 ng/mL, respectively. As a result, the content of oleanolic acid was the highest extracted by ultrasonic extraction and the content of mangiferin was the highest extracted by reflux extraction. For percolation extraction, the contents of five components were between ultrasound and reflux extraction. Conclusion: For five components, there are significant differences between the three different extraction methods. The results could provide a reference for the quality control of Swertia mussotii Franch and the research and development of new drugs.

  5. Texture analysis with statistical methods for wheat ear extraction

    NASA Astrophysics Data System (ADS)

    Bakhouche, M.; Cointault, F.; Gouton, P.

    2007-01-01

    In agronomic domain, the simplification of crop counting, necessary for yield prediction and agronomic studies, is an important project for technical institutes such as Arvalis. Although the main objective of our global project is to conceive a mobile robot for natural image acquisition directly in a field, Arvalis has proposed us first to detect by image processing the number of wheat ears in images before to count them, which will allow to obtain the first component of the yield. In this paper we compare different texture image segmentation techniques based on feature extraction by first and higher order statistical methods which have been applied on our images. The extracted features are used for unsupervised pixel classification to obtain the different classes in the image. So, the K-means algorithm is implemented before the choice of a threshold to highlight the ears. Three methods have been tested in this feasibility study with very average error of 6%. Although the evaluation of the quality of the detection is visually done, automatic evaluation algorithms are currently implementing. Moreover, other statistical methods of higher order will be implemented in the future jointly with methods based on spatio-frequential transforms and specific filtering.

  6. An Overview of the Computational Physics and Methods Group at Los Alamos National Laboratory

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

    Baker, Randal Scott

    CCS Division was formed to strengthen the visibility and impact of computer science and computational physics research on strategic directions for the Laboratory. Both computer science and computational science are now central to scientific discovery and innovation. They have become indispensable tools for all other scientific missions at the Laboratory. CCS Division forms a bridge between external partners and Laboratory programs, bringing new ideas and technologies to bear on today’s important problems and attracting high-quality technical staff members to the Laboratory. The Computational Physics and Methods Group CCS-2 conducts methods research and develops scientific software aimed at the latest andmore » emerging HPC systems.« less

  7. Method for liquid chromatographic extraction of strontium from acid solutions

    DOEpatents

    Horwitz, E. Philip; Dietz, Mark L.

    1992-01-01

    A method and apparatus for extracting strontium and technetium values from biological, industrial and environmental sample solutions using a chromatographic column is described. An extractant medium for the column is prepared by generating a solution of a diluent containing a Crown ether and dispersing the solution on a resin substrate material. The sample solution is highly acidic and is introduced directed to the chromatographic column and strontium or technetium is eluted using deionized water.

  8. Comparison of Two Simplification Methods for Shoreline Extraction from Digital Orthophoto Images

    NASA Astrophysics Data System (ADS)

    Bayram, B.; Sen, A.; Selbesoglu, M. O.; Vārna, I.; Petersons, P.; Aykut, N. O.; Seker, D. Z.

    2017-11-01

    The coastal ecosystems are very sensitive to external influences. Coastal resources such as sand dunes, coral reefs and mangroves has vital importance to prevent coastal erosion. Human based effects also threats the coastal areas. Therefore, the change of coastal areas should be monitored. Up-to-date, accurate shoreline information is indispensable for coastal managers and decision makers. Remote sensing and image processing techniques give a big opportunity to obtain reliable shoreline information. In the presented study, NIR bands of seven 1:5000 scaled digital orthophoto images of Riga Bay-Latvia have been used. The Object-oriented Simple Linear Clustering method has been utilized to extract shoreline of Riga Bay. Bend and Douglas-Peucker methods have been used to simplify the extracted shoreline to test the effect of both methods. Photogrammetrically digitized shoreline has been taken as reference data to compare obtained results. The accuracy assessment has been realised by Digital Shoreline Analysis tool. As a result, the achieved shoreline by the Bend method has been found closer to the extracted shoreline with Simple Linear Clustering method.

  9. A Method for Extracting Road Boundary Information from Crowdsourcing Vehicle GPS Trajectories.

    PubMed

    Yang, Wei; Ai, Tinghua; Lu, Wei

    2018-04-19

    Crowdsourcing trajectory data is an important approach for accessing and updating road information. In this paper, we present a novel approach for extracting road boundary information from crowdsourcing vehicle traces based on Delaunay triangulation (DT). First, an optimization and interpolation method is proposed to filter abnormal trace segments from raw global positioning system (GPS) traces and interpolate the optimization segments adaptively to ensure there are enough tracking points. Second, constructing the DT and the Voronoi diagram within interpolated tracking lines to calculate road boundary descriptors using the area of Voronoi cell and the length of triangle edge. Then, the road boundary detection model is established integrating the boundary descriptors and trajectory movement features (e.g., direction) by DT. Third, using the boundary detection model to detect road boundary from the DT constructed by trajectory lines, and a regional growing method based on seed polygons is proposed to extract the road boundary. Experiments were conducted using the GPS traces of taxis in Beijing, China, and the results show that the proposed method is suitable for extracting the road boundary from low-frequency GPS traces, multi-type road structures, and different time intervals. Compared with two existing methods, the automatically extracted boundary information was proved to be of higher quality.

  10. A Method for Extracting Road Boundary Information from Crowdsourcing Vehicle GPS Trajectories

    PubMed Central

    Yang, Wei

    2018-01-01

    Crowdsourcing trajectory data is an important approach for accessing and updating road information. In this paper, we present a novel approach for extracting road boundary information from crowdsourcing vehicle traces based on Delaunay triangulation (DT). First, an optimization and interpolation method is proposed to filter abnormal trace segments from raw global positioning system (GPS) traces and interpolate the optimization segments adaptively to ensure there are enough tracking points. Second, constructing the DT and the Voronoi diagram within interpolated tracking lines to calculate road boundary descriptors using the area of Voronoi cell and the length of triangle edge. Then, the road boundary detection model is established integrating the boundary descriptors and trajectory movement features (e.g., direction) by DT. Third, using the boundary detection model to detect road boundary from the DT constructed by trajectory lines, and a regional growing method based on seed polygons is proposed to extract the road boundary. Experiments were conducted using the GPS traces of taxis in Beijing, China, and the results show that the proposed method is suitable for extracting the road boundary from low-frequency GPS traces, multi-type road structures, and different time intervals. Compared with two existing methods, the automatically extracted boundary information was proved to be of higher quality. PMID:29671792

  11. Optimization of extractants, purifying packings, and eluents for analytical extraction of organochlorine pesticides in Hydragric Acrisols.

    PubMed

    Zhao, Zhenhua; Xia, Liling; Wang, Fang; Jiang, Xin; Gao, Yanzheng

    2012-08-01

    In this study, we screened for an economic, rapid, and efficient hypotoxic pretreatment method for organochlorine pesticides in soil samples for gas chromatography (GC) analysis. The analytical extraction efficiencies of 11 different extractants, nine types of solid-phase purification (SPP) cartridge packings, and three types of eluents for 13 organochlorine pesticides (OCPs) in spiked and natural Chinese red soil (Hydragric Acrisols) were evaluated using an ultrasonic extraction and solid-phase purification method. High percent recoveries (85-106%) were obtained for the 13 organochlorine pesticides in soil when petroleum ether/acetone/water (10:5:2, v/v) was used an extractant. They were purified using celite SPP cartridge packing and eluted with 9 mL of dichloromethane/petroleum ether (1:9, v/v). The OCPs purification pretreatment of Hydragric Acrisols, using the above method, meets the GC analysis requirements. Compared with other traditional pretreatment methods for OCPs in soil samples, this method has several advantages, such as a short extraction time, reducing the amount of solvent, having no emulsion phenomenon, and hypotoxicity to the laboratory technicians. The concentrations of 1,1,1,-trichloro-2(p-chlorophenyl)-2-(o-chlorophenyl) ethane (DDTs; 3.42-8.08 ng g(-1)) in field soils were higher than the hexachlorocyclohexane concentration (2.94-6.12 ng g(-1)). The 1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (p,p'-DDE) + 1,1-dichloro-2,2-bis(p-chlorophenyl)-ethane (p,p'-DDD)/p,p'-DDT ratio in this field soil was approximately 2.7, suggesting that no new DDT pollution source was introduced into the sampling site.

  12. Modified Lipid Extraction Methods for Deep Subsurface Shale

    PubMed Central

    Akondi, Rawlings N.; Trexler, Ryan V.; Pfiffner, Susan M.; Mouser, Paula J.; Sharma, Shikha

    2017-01-01

    Growing interest in the utilization of black shales for hydrocarbon development and environmental applications has spurred investigations of microbial functional diversity in the deep subsurface shale ecosystem. Lipid biomarker analyses including phospholipid fatty acids (PLFAs) and diglyceride fatty acids (DGFAs) represent sensitive tools for estimating biomass and characterizing the diversity of microbial communities. However, complex shale matrix properties create immense challenges for microbial lipid extraction procedures. Here, we test three different lipid extraction methods: modified Bligh and Dyer (mBD), Folch (FOL), and microwave assisted extraction (MAE), to examine their ability in the recovery and reproducibility of lipid biomarkers in deeply buried shales. The lipid biomarkers were analyzed as fatty acid methyl esters (FAMEs) with the GC-MS, and the average PL-FAME yield ranged from 67 to 400 pmol/g, while the average DG-FAME yield ranged from 600 to 3,000 pmol/g. The biomarker yields in the intact phospholipid Bligh and Dyer treatment (mBD + Phos + POPC), the Folch, the Bligh and Dyer citrate buffer (mBD-Cit), and the MAE treatments were all relatively higher and statistically similar compared to the other extraction treatments for both PLFAs and DGFAs. The biomarker yields were however highly variable within replicates for most extraction treatments, although the mBD + Phos + POPC treatment had relatively better reproducibility in the consistent fatty acid profiles. This variability across treatments which is associated with the highly complex nature of deeply buried shale matrix, further necessitates customized methodological developments for the improvement of lipid biomarker recovery. PMID:28790998

  13. Modular workcells: modern methods for laboratory automation.

    PubMed

    Felder, R A

    1998-12-01

    Laboratory automation is beginning to become an indispensable survival tool for laboratories facing difficult market competition. However, estimates suggest that only 8% of laboratories will be able to afford total laboratory automation systems. Therefore, automation vendors have developed alternative hardware configurations called 'modular automation', to fit the smaller laboratory. Modular automation consists of consolidated analyzers, integrated analyzers, modular workcells, and pre- and post-analytical automation. These terms will be defined in this paper. Using a modular automation model, the automated core laboratory will become a site where laboratory data is evaluated by trained professionals to provide diagnostic information to practising physicians. Modem software information management and process control tools will complement modular hardware. Proper standardization that will allow vendor-independent modular configurations will assure success of this revolutionary new technology.

  14. Laboratory Validation and Field Assessment of Petroleum Laboratory Technicians' Dermal Exposure to Crude Oil Using a Wipe Sampling Method.

    PubMed

    Galea, Karen S; Mueller, Will; Arfaj, Ayman M; Llamas, Jose L; Buick, Jennifer; Todd, David; McGonagle, Carolyn

    2018-05-21

    Crude oil may cause adverse dermal effects therefore dermal exposure is an exposure route of concern. Galea et al. (2014b) reported on a study comparing recovery (wipe) and interception (cotton glove) dermal sampling methods. The authors concluded that both methods were suitable for assessing dermal exposure to oil-based drilling fluids and crude oil but that glove samplers may overestimate the amount of fluid transferred to the skin. We describe a study which aimed to further evaluate the wipe sampling method to assess dermal exposure to crude oil, with this assessment including extended sample storage periods and sampling efficiency tests being undertaken at environmental conditions to mimic those typical of outdoor conditions in Saudi Arabia. The wipe sampling method was then used to assess the laboratory technicians' actual exposure to crude oil during typical petroleum laboratory tasks. Overall, acceptable storage efficiencies up to 54 days were reported with results suggesting storage stability over time. Sampling efficiencies were also reported to be satisfactory at both ambient and elevated temperature and relative humidity environmental conditions for surrogate skin spiked with known masses of crude oil and left up to 4 h prior to wiping, though there was an indication of reduced sampling efficiency over time. Nineteen petroleum laboratory technicians provided a total of 35 pre- and 35 post-activity paired hand wipe samples. Ninety-three percent of the pre-exposure paired hand wipes were less than the analytical limit of detection (LOD), whereas 46% of the post-activity paired hand wipes were less than the LOD. The geometric mean paired post-activity wipe sample measurement was 3.09 µg cm-2 (range 1.76-35.4 µg cm-2). It was considered that dermal exposure most frequently occurred through direct contact with the crude oil (emission) or via deposition. The findings of this study suggest that the wipe sampling method is satisfactory in quantifying

  15. A method for the extraction and quantitation of phycoerythrin from algae

    NASA Technical Reports Server (NTRS)

    Stewart, D. E.

    1982-01-01

    A summary of a new technique for the extraction and quantitation of phycoerythrin (PHE) from algal samples is described. Results of analysis of four extracts representing three PHE types from algae including cryptomonad and cyanophyte types are presented. The method of extraction and an equation for quantitation are given. A graph showing the relationship of concentration and fluorescence units that may be used with samples fluorescing around 575-580 nm (probably dominated by cryptophytes in estuarine waters) and 560 nm (dominated by cyanophytes characteristics of the open ocean) is provided.

  16. Study Unveils New Method for Universal Extraction and PCR Amplification of Fungal DNA

    DTIC Science & Technology

    2014-06-12

    Wickes noted that there are methods to extract fungi from soil , for example, and "once you get down to pure DNA, everything else is the same," he said...rare or hard to identify fungal infections. The new extraction and amplification method can be universally applied to fungi , according to the...best treatments. In addition, rare fungi , or species with phenotypic doppelgangers, can stump medical mycologists, so molecular methods are critical

  17. An efficient field and laboratory workflow for plant phylotranscriptomic projects1

    PubMed Central

    Yang, Ya; Moore, Michael J.; Brockington, Samuel F.; Timoneda, Alfonso; Feng, Tao; Marx, Hannah E.; Walker, Joseph F.; Smith, Stephen A.

    2017-01-01

    Premise of the study: We describe a field and laboratory workflow developed for plant phylotranscriptomic projects that involves cryogenic tissue collection in the field, RNA extraction and quality control, and library preparation. We also make recommendations for sample curation. Methods and Results: A total of 216 frozen tissue samples of Caryophyllales and other angiosperm taxa were collected from the field or botanical gardens. RNA was extracted, stranded mRNA libraries were prepared, and libraries were sequenced on Illumina HiSeq platforms. These included difficult mucilaginous tissues such as those of Cactaceae and Droseraceae. Conclusions: Our workflow is not only cost effective (ca. $270 per sample, as of August 2016, from tissue to reads) and time efficient (less than 50 h for 10–12 samples including all laboratory work and sample curation), but also has proven robust for extraction of difficult samples such as tissues containing high levels of secondary compounds. PMID:28337391

  18. The current role of on-line extraction approaches in clinical and forensic toxicology.

    PubMed

    Mueller, Daniel M

    2014-08-01

    In today's clinical and forensic toxicological laboratories, automation is of interest because of its ability to optimize processes, to reduce manual workload and handling errors and to minimize exposition to potentially infectious samples. Extraction is usually the most time-consuming step; therefore, automation of this step is reasonable. Currently, from the field of clinical and forensic toxicology, methods using the following on-line extraction techniques have been published: on-line solid-phase extraction, turbulent flow chromatography, solid-phase microextraction, microextraction by packed sorbent, single-drop microextraction and on-line desorption of dried blood spots. Most of these published methods are either single-analyte or multicomponent procedures; methods intended for systematic toxicological analysis are relatively scarce. However, the use of on-line extraction will certainly increase in the near future.

  19. Comparison of accelerated solvent extraction and quick, easy, cheap, effective, rugged and safe method for extraction and determination of pharmaceuticals in vegetables.

    PubMed

    Chuang, Ya-Hui; Zhang, Yingjie; Zhang, Wei; Boyd, Stephen A; Li, Hui

    2015-07-24

    Land application of biosolids and irrigation with reclaimed water in agricultural production could result in accumulation of pharmaceuticals in vegetable produce. To better assess the potential human health impact from long-term consumption of pharmaceutical-contaminated vegetables, it is important to accurately quantify the amount of pharmaceuticals accumulated in vegetables. In this study, a quick, easy, cheap, effective, rugged and safe (QuEChERS) method was developed and optimized to extract multiple classes of pharmaceuticals from vegetables, which were subsequently quantified by liquid chromatography coupled to tandem mass spectrometry. For the eleven target pharmaceuticals in celery and lettuce, the extraction recovery of the QuEChERS method ranged from 70.1 to 118.6% with relative standard deviation <20%, and the method detection limit was achieved at the levels of nanograms of pharmaceuticals per gram of vegetables. The results revealed that the performance of the QuEChERS method was comparable to, or better than that of accelerated solvent extraction (ASE) method for extraction of pharmaceuticals from plants. The two optimized extraction methods were applied to quantify the uptake of pharmaceuticals by celery and lettuce growing hydroponically. The results showed that all the eleven target pharmaceuticals could be absorbed by the vegetables from water. Compared to the ASE method, the QuEChERS method offers the advantages of short time and reduced costs of sample preparation, and less amount of organic solvents used. The established QuEChERS method could be used to determine the accumulation of multiple classes of pharmaceutical residues in vegetables and other plants, which is needed to evaluate the quality and safety of agricultural produce consumed by humans. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Extraction and analysis of cortisol from human and monkey hair.

    PubMed

    Meyer, Jerrold; Novak, Melinda; Hamel, Amanda; Rosenberg, Kendra

    2014-01-24

    The stress hormone cortisol (CORT) is slowly incorporated into the growing hair shaft of humans, nonhuman primates, and other mammals. We developed and validated a method for CORT extraction and analysis from rhesus monkey hair and subsequently adapted this method for use with human scalp hair. In contrast to CORT "point samples" obtained from plasma or saliva, hair CORT provides an integrated measure of hypothalamic-pituitary-adrenocortical (HPA) system activity, and thus physiological stress, during the period of hormone incorporation. Because human scalp hair grows at an average rate of 1 cm/month, CORT levels obtained from hair segments several cm in length can potentially serve as a biomarker of stress experienced over a number of months. In our method, each hair sample is first washed twice in isopropanol to remove any CORT from the outside of the hair shaft that has been deposited from sweat or sebum. After drying, the sample is ground to a fine powder to break up the hair's protein matrix and increase the surface area for extraction. CORT from the interior of the hair shaft is extracted into methanol, the methanol is evaporated, and the extract is reconstituted in assay buffer. Extracted CORT, along with standards and quality controls, is then analyzed by means of a sensitive and specific commercially available enzyme immunoassay (EIA) kit. Readout from the EIA is converted to pg CORT per mg powdered hair weight. This method has been used in our laboratory to analyze hair CORT in humans, several species of macaque monkeys, marmosets, dogs, and polar bears. Many studies both from our lab and from other research groups have demonstrated the broad applicability of hair CORT for assessing chronic stress exposure in natural as well as laboratory settings.

  1. A method for real-time implementation of HOG feature extraction

    NASA Astrophysics Data System (ADS)

    Luo, Hai-bo; Yu, Xin-rong; Liu, Hong-mei; Ding, Qing-hai

    2011-08-01

    Histogram of oriented gradient (HOG) is an efficient feature extraction scheme, and HOG descriptors are feature descriptors which is widely used in computer vision and image processing for the purpose of biometrics, target tracking, automatic target detection(ATD) and automatic target recognition(ATR) etc. However, computation of HOG feature extraction is unsuitable for hardware implementation since it includes complicated operations. In this paper, the optimal design method and theory frame for real-time HOG feature extraction based on FPGA were proposed. The main principle is as follows: firstly, the parallel gradient computing unit circuit based on parallel pipeline structure was designed. Secondly, the calculation of arctangent and square root operation was simplified. Finally, a histogram generator based on parallel pipeline structure was designed to calculate the histogram of each sub-region. Experimental results showed that the HOG extraction can be implemented in a pixel period by these computing units.

  2. Characterization of rice starch and protein obtained by a fast alkaline extraction method.

    PubMed

    Souza, Daiana de; Sbardelotto, Arthur Francisco; Ziegler, Denize Righetto; Marczak, Ligia Damasceno Ferreira; Tessaro, Isabel Cristina

    2016-01-15

    This study evaluated the characteristics of rice starch and protein obtained by a fast alkaline extraction method on rice flour (RF) derived from broken rice. The extraction was conducted using 0.18% NaOH at 30°C for 30min followed by centrifugation to separate the starch rich and the protein rich fractions. This fast extraction method allowed to obtain an isoelectric precipitation protein concentrate (IPPC) with 79% protein and a starchy product with low protein content. The amino acid content of IPPC was practically unchanged compared to the protein in RF. The proteins of the IPPC underwent denaturation during extraction and some of the starch suffered the cold gelatinization phenomenon, due to the alkaline treatment. With some modifications, the fast method can be interesting in a technological point of view as it enables process cost reduction and useful ingredients obtention to the food and chemical industries. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. A comparative study of Averrhoabilimbi extraction method

    NASA Astrophysics Data System (ADS)

    Zulhaimi, H. I.; Rosli, I. R.; Kasim, K. F.; Akmal, H. Muhammad; Nuradibah, M. A.; Sam, S. T.

    2017-09-01

    In recent year, bioactive compound in plant has become a limelight in the food and pharmaceutical market, leading to research interest to implement effective technologies for extracting bioactive substance. Therefore, this study is focusing on extraction of Averrhoabilimbi by different extraction technique namely, maceration and ultrasound-assisted extraction. Fewplant partsof Averrhoabilimbiweretaken as extraction samples which are fruits, leaves and twig. Different solvents such as methanol, ethanol and distilled water were utilized in the process. Fruit extractsresult in highest extraction yield compared to other plant parts. Ethanol and distilled water have significant role compared to methanol in all parts and both extraction technique. The result also shows that ultrasound-assisted extraction gave comparable result with maceration. Besides, the shorter period on extraction process gives useful in term of implementation to industries.

  4. Insights from a Modular Interdisciplinary Laboratory Course

    NASA Astrophysics Data System (ADS)

    Bouvier-Brown, N. C.; Carmona, V.

    2016-12-01

    A laboratory curriculum is naturally oriented towards "hands on" and problem-based learning. An earth science course, like chemical ecology, has the additional advantage of interdisciplinary education. Our Chemical Ecology course at Loyola Marymount University was structured as a small modular workshop. Students first gained hands-on experience with each analytical technique in the laboratory. The class then discussed these first datasets, delved into lecture topics, and tweaked their experimental procedures. Lastly, students were given time to design and execute their own experiments and present their findings. Three-to-four class periods were allotted for each laboratory topic. Detailed information and student reflections will be presented from the Spectroscopy module. Students used the Folin-Ciocalteau reagent method to extract the phenolic content of vegetation and/or soils. Because phenols are produced by plants for defense, this spectroscopic laboratory activity provided insight on allelopathy using analytical chemistry. Students were extremely engaged and learned not only the lab theory and technique, but also its application to our local ecology.

  5. SW-846 Test Method 3200: Mercury Species Fractionation and Quantification by Microwave Assisted Extraction, Selective Solvent Extraction and/or Solid Phase Extraction

    EPA Pesticide Factsheets

    a sequential extraction and separation procedure that maybe used in conjunction with a determinative method to differentiate mercury species that arepresent in soils and sediments. provides information on both total mercury andvarious mercury species.

  6. A new method of tree xylem water extraction for isotopic analysis

    NASA Astrophysics Data System (ADS)

    Gierke, C.; Newton, B. T.

    2011-12-01

    The Sacramento Mountain Watershed Study in the southern Sacramento Mountains of New Mexico is designed to assess the forest restoration technique of tree thinning in mountain watersheds as an effective method of increasing local and regional groundwater recharge. The project is using a soil water balance approach to quantify the partitioning of local precipitation within this watershed before and after thinning trees. Understanding what sources trees extract their water from (e.g. shallow groundwater, unsaturated fractured bedrock, and soils) is difficult due to a complex hydrologic system and heterogeneous distribution of soil thicknesses. However, in order to accurately quantify the soil water balance and to assess how thinning trees will affect this water balance, it is important determine the sources from which trees extract their water. We plan to use oxygen and hydrogen stable isotopic analysis of various end member waters to identify these different sources. We are in the process of developing a new method of determining the isotopic composition of tree water that has several advantages over conventional methods. Within the tree there is the xylem which transports water from the roots to the leaves and the phloem which transports starches and sugars in a water media throughout the tree. Previous studies have shown that the isotopic composition of xylem water accurately reflects that of source water, while phloem water has undergone isotopic fractionation during photosynthesis and metabolism. The distillation of water from twigs, which is often used to extract tree water for isotopic analysis, is very labor intensive. Other disadvantages to distillation methods include possible fractionation due to phase changes and the possible extraction of fractionated phloem waters. Employing a new mixing method, the composition of the twig water (TW) can be determined by putting twigs of unknown isotopic water composition into waters of known compositions or initial

  7. Mechanomyographic Parameter Extraction Methods: An Appraisal for Clinical Applications

    PubMed Central

    Ibitoye, Morufu Olusola; Hamzaid, Nur Azah; Zuniga, Jorge M.; Hasnan, Nazirah; Wahab, Ahmad Khairi Abdul

    2014-01-01

    The research conducted in the last three decades has collectively demonstrated that the skeletal muscle performance can be alternatively assessed by mechanomyographic signal (MMG) parameters. Indices of muscle performance, not limited to force, power, work, endurance and the related physiological processes underlying muscle activities during contraction have been evaluated in the light of the signal features. As a non-stationary signal that reflects several distinctive patterns of muscle actions, the illustrations obtained from the literature support the reliability of MMG in the analysis of muscles under voluntary and stimulus evoked contractions. An appraisal of the standard practice including the measurement theories of the methods used to extract parameters of the signal is vital to the application of the signal during experimental and clinical practices, especially in areas where electromyograms are contraindicated or have limited application. As we highlight the underpinning technical guidelines and domains where each method is well-suited, the limitations of the methods are also presented to position the state of the art in MMG parameters extraction, thus providing the theoretical framework for improvement on the current practices to widen the opportunity for new insights and discoveries. Since the signal modality has not been widely deployed due partly to the limited information extractable from the signals when compared with other classical techniques used to assess muscle performance, this survey is particularly relevant to the projected future of MMG applications in the realm of musculoskeletal assessments and in the real time detection of muscle activity. PMID:25479326

  8. Evaluation of Lysis Methods for the Extraction of Bacterial DNA for Analysis of the Vaginal Microbiota

    PubMed Central

    Gill, Christina; Blow, Frances; Darby, Alistair C.

    2016-01-01

    Background Recent studies on the vaginal microbiota have employed molecular techniques such as 16S rRNA gene sequencing to describe the bacterial community as a whole. These techniques require the lysis of bacterial cells to release DNA before purification and PCR amplification of the 16S rRNA gene. Currently, methods for the lysis of bacterial cells are not standardised and there is potential for introducing bias into the results if some bacterial species are lysed less efficiently than others. This study aimed to compare the results of vaginal microbiota profiling using four different pretreatment methods for the lysis of bacterial samples (30 min of lysis with lysozyme, 16 hours of lysis with lysozyme, 60 min of lysis with a mixture of lysozyme, mutanolysin and lysostaphin and 30 min of lysis with lysozyme followed by bead beating) prior to chemical and enzyme-based DNA extraction with a commercial kit. Results After extraction, DNA yield did not significantly differ between methods with the exception of lysis with lysozyme combined with bead beating which produced significantly lower yields when compared to lysis with the enzyme cocktail or 30 min lysis with lysozyme only. However, this did not result in a statistically significant difference in the observed alpha diversity of samples. The beta diversity (Bray-Curtis dissimilarity) between different lysis methods was statistically significantly different, but this difference was small compared to differences between samples, and did not affect the grouping of samples with similar vaginal bacterial community structure by hierarchical clustering. Conclusions An understanding of how laboratory methods affect the results of microbiota studies is vital in order to accurately interpret the results and make valid comparisons between studies. Our results indicate that the choice of lysis method does not prevent the detection of effects relating to the type of vaginal bacterial community one of the main outcome measures

  9. Evaluation of Lysis Methods for the Extraction of Bacterial DNA for Analysis of the Vaginal Microbiota.

    PubMed

    Gill, Christina; van de Wijgert, Janneke H H M; Blow, Frances; Darby, Alistair C

    2016-01-01

    Recent studies on the vaginal microbiota have employed molecular techniques such as 16S rRNA gene sequencing to describe the bacterial community as a whole. These techniques require the lysis of bacterial cells to release DNA before purification and PCR amplification of the 16S rRNA gene. Currently, methods for the lysis of bacterial cells are not standardised and there is potential for introducing bias into the results if some bacterial species are lysed less efficiently than others. This study aimed to compare the results of vaginal microbiota profiling using four different pretreatment methods for the lysis of bacterial samples (30 min of lysis with lysozyme, 16 hours of lysis with lysozyme, 60 min of lysis with a mixture of lysozyme, mutanolysin and lysostaphin and 30 min of lysis with lysozyme followed by bead beating) prior to chemical and enzyme-based DNA extraction with a commercial kit. After extraction, DNA yield did not significantly differ between methods with the exception of lysis with lysozyme combined with bead beating which produced significantly lower yields when compared to lysis with the enzyme cocktail or 30 min lysis with lysozyme only. However, this did not result in a statistically significant difference in the observed alpha diversity of samples. The beta diversity (Bray-Curtis dissimilarity) between different lysis methods was statistically significantly different, but this difference was small compared to differences between samples, and did not affect the grouping of samples with similar vaginal bacterial community structure by hierarchical clustering. An understanding of how laboratory methods affect the results of microbiota studies is vital in order to accurately interpret the results and make valid comparisons between studies. Our results indicate that the choice of lysis method does not prevent the detection of effects relating to the type of vaginal bacterial community one of the main outcome measures of epidemiological studies

  10. A COMPARISON OF AUTOMATED AND TRADITIONAL METHODS FOR THE EXTRACTION OF ARSENICALS FROM FISH

    EPA Science Inventory

    An automated extractor employing accelerated solvent extraction (ASE) has been compared with a traditional sonication method of extraction for the extraction of arsenicals from fish tissue. Four different species of fish and a standard reference material, DORM-2, were subjected t...

  11. Social network extraction based on Web: 3. the integrated superficial method

    NASA Astrophysics Data System (ADS)

    Nasution, M. K. M.; Sitompul, O. S.; Noah, S. A.

    2018-03-01

    The Web as a source of information has become part of the social behavior information. Although, by involving only the limitation of information disclosed by search engines in the form of: hit counts, snippets, and URL addresses of web pages, the integrated extraction method produces a social network not only trusted but enriched. Unintegrated extraction methods may produce social networks without explanation, resulting in poor supplemental information, or resulting in a social network of durmise laden, consequently unrepresentative social structures. The integrated superficial method in addition to generating the core social network, also generates an expanded network so as to reach the scope of relation clues, or number of edges computationally almost similar to n(n - 1)/2 for n social actors.

  12. Efficacy Evaluation of Different Wavelet Feature Extraction Methods on Brain MRI Tumor Detection

    NASA Astrophysics Data System (ADS)

    Nabizadeh, Nooshin; John, Nigel; Kubat, Miroslav

    2014-03-01

    Automated Magnetic Resonance Imaging brain tumor detection and segmentation is a challenging task. Among different available methods, feature-based methods are very dominant. While many feature extraction techniques have been employed, it is still not quite clear which of feature extraction methods should be preferred. To help improve the situation, we present the results of a study in which we evaluate the efficiency of using different wavelet transform features extraction methods in brain MRI abnormality detection. Applying T1-weighted brain image, Discrete Wavelet Transform (DWT), Discrete Wavelet Packet Transform (DWPT), Dual Tree Complex Wavelet Transform (DTCWT), and Complex Morlet Wavelet Transform (CMWT) methods are applied to construct the feature pool. Three various classifiers as Support Vector Machine, K Nearest Neighborhood, and Sparse Representation-Based Classifier are applied and compared for classifying the selected features. The results show that DTCWT and CMWT features classified with SVM, result in the highest classification accuracy, proving of capability of wavelet transform features to be informative in this application.

  13. Comparison of different methods for extraction from Tetraclinis articulata: yield, chemical composition and antioxidant activity.

    PubMed

    Herzi, Nejia; Bouajila, Jalloul; Camy, Séverine; Romdhane, Mehrez; Condoret, Jean-Stéphane

    2013-12-15

    In the present study, three techniques of extraction: hydrodistillation (HD), solvent extraction (conventional 'Soxhlet' technique) and an innovative technique, i.e., the supercritical fluid extraction (SFE), were applied to ground Tetraclinis articulata leaves and compared for extraction duration, extraction yield, and chemical composition of the extracts as well as their antioxidant activities. The extracts were analyzed by GC-FID and GC-MS. The antioxidant activity was measured using two methods: ABTS(•+) and DPPH(•). The yield obtained using HD, SFE, hexane and ethanol Soxhlet extractions were found to be 0.6, 1.6, 40.4 and 21.2-27.4 g/kg respectively. An original result of this study is that the best antioxidant activity was obtained with an SFE extract (41 mg/L). The SFE method offers some noteworthy advantages over traditional alternatives, such as shorter extraction times, low environmental impact, and a clean, non-thermally-degraded final product. Also, a good correlation between the phenolic contents and the antioxidant activity was observed with extracts obtained by SFE at 9 MPa. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. A Low-Storage-Consumption XML Labeling Method for Efficient Structural Information Extraction

    NASA Astrophysics Data System (ADS)

    Liang, Wenxin; Takahashi, Akihiro; Yokota, Haruo

    Recently, labeling methods to extract and reconstruct the structural information of XML data, which are important for many applications such as XPath query and keyword search, are becoming more attractive. To achieve efficient structural information extraction, in this paper we propose C-DO-VLEI code, a novel update-friendly bit-vector encoding scheme, based on register-length bit operations combining with the properties of Dewey Order numbers, which cannot be implemented in other relevant existing schemes such as ORDPATH. Meanwhile, the proposed method also achieves lower storage consumption because it does not require either prefix schema or any reserved codes for node insertion. We performed experiments to evaluate and compare the performance and storage consumption of the proposed method with those of the ORDPATH method. Experimental results show that the execution times for extracting depth information and parent node labels using the C-DO-VLEI code are about 25% and 15% less, respectively, and the average label size using the C-DO-VLEI code is about 24% smaller, comparing with ORDPATH.

  15. An improved high-throughput lipid extraction method for the analysis of human brain lipids.

    PubMed

    Abbott, Sarah K; Jenner, Andrew M; Mitchell, Todd W; Brown, Simon H J; Halliday, Glenda M; Garner, Brett

    2013-03-01

    We have developed a protocol suitable for high-throughput lipidomic analysis of human brain samples. The traditional Folch extraction (using chloroform and glass-glass homogenization) was compared to a high-throughput method combining methyl-tert-butyl ether (MTBE) extraction with mechanical homogenization utilizing ceramic beads. This high-throughput method significantly reduced sample handling time and increased efficiency compared to glass-glass homogenizing. Furthermore, replacing chloroform with MTBE is safer (less carcinogenic/toxic), with lipids dissolving in the upper phase, allowing for easier pipetting and the potential for automation (i.e., robotics). Both methods were applied to the analysis of human occipital cortex. Lipid species (including ceramides, sphingomyelins, choline glycerophospholipids, ethanolamine glycerophospholipids and phosphatidylserines) were analyzed via electrospray ionization mass spectrometry and sterol species were analyzed using gas chromatography mass spectrometry. No differences in lipid species composition were evident when the lipid extraction protocols were compared, indicating that MTBE extraction with mechanical bead homogenization provides an improved method for the lipidomic profiling of human brain tissue.

  16. [Laboratory diagnosis of genital herpes--direct immunofluorescence method].

    PubMed

    Majewska, Anna; Romejko-Wolniewicz, Ewa; Zareba-Szczudlik, Julia; Kilijańczyk, Marek; Gajewska, Małgorzata; Młynarczyk, Grazyna

    2013-07-01

    Aim of the study was to determine clinical usefulness of direct immunofluorescence method in the laboratory diagnosis of genital herpes in women. Overall 187 anogenital swabs were collected from 120 women. Using a dacron-tipped applicator 83 swabs were collected from women suspected of genital herpes and 104 from patients with no signs of genital infection. All samples were tested using cell culture (Vero cell line) and then direct immunofluorescence method (DIF) for the identification of antigens of herpes simplex viruses: HSV-1 and HSV-2. Characteristic cytopathic effect (CPE), indicative of alphaherpesvirus infection, was observed in 43.4% of cultures with clinical specimens collected from women with suspected genital herpes and in 29.8% of cultures of clinical specimens taken from patients with no clinical symptoms of genital herpes. Herpes simplex viruses were determined in 73 samples by direct immunofluorescence method after amplification of the virus in cell culture. The DIF test confirmed the diagnosis based on the microscopic CPE observation in 85%. In 15% of samples (taken from pregnant women without clinical signs of infection) we reported positive immunofluorescence in the absence of CPE. The frequency of antigen detection was statistically significantly higher in samples that were positive by culture study (chi-square test with Yates's correction, p < 0.01). This method proved to be highly sensitive (97%) in women with clinically suspected infection. High negative predictive value (99%) proves the clinical utility of the DIF in these group of patients. In asymptomatic infections, viral antigens were detected most frequently in the swabs from the cervical canal, and in cases of suspected genital herpes in swabs taken from the vestibule of the vagina and the vulva. However, there was no statistically significant difference in the frequency of detection of Herpes Simplex Virus antigens in specimens from different parts of the genital tract in both groups

  17. An effective placental cotyledons proteins extraction method for 2D gel electrophoresis.

    PubMed

    Tan, Niu J; Daim, Leona D J; Jamil, Amilia A M; Mohtarrudin, Norhafizah; Thilakavathy, Karuppiah

    2017-03-01

    Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this study is to determine the best protein extraction protocol for the extraction of protein from placental cotyledons tissues for a two-dimensional gel electrophoresis (2D-GE). Based on widely used protein extraction strategies, 12 different extraction methodologies were carefully selected, which included one chemical extraction, two mechanical extraction coupled protein precipitations, and nine chemical extraction coupled protein precipitations. Extracted proteins were resolved in a one-dimensional gel electrophoresis and 2D-GE; then, it was compared with set criteria: extraction efficacy, protein resolution, reproducibility, and recovery efficiency. Our results revealed that a better profile was obtained by chemical extraction in comparison to mechanical extraction. We further compared chemical extraction coupled protein precipitation methodologies, where the DNase/lithium chloride-dense sucrose homogenization coupled dichloromethane-methanol precipitation (DNase/LiCl-DSH-D/MPE) method showed good protein extraction efficiency. This, however, was carried out with the best protein resolution and proteome reproducibility on 2D-gels. DNase/LiCl-DSH-D/MPE was efficient in the extraction of proteins from placental cotyledons tissues. In addition, this methodology could hypothetically allow the protein extraction of any tissue that contains highly abundant lipid and glycogen. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Method 1664, Revision A: n-hexane extractable material (HEM; oil and grease) and silica gel treated n-hexane extractable material (SGT-HEM; non-polar material) by extraction and gravimetry

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

    Not Available

    This method is for determination of n-hexane extractable material (HEM; oil and grease) and n-hexane extractable material that is not adsorbed by silica gel (SGT-HEM; non-polar material) in surface and saline waters and industrial and domestic aqueous wastes. Extractable materials that may be determined are relatively non-volatile hydrocarbons, vegetable oils, animal fats, waxes, soaps, greases, and related materials. This method is capable of measuring HEM and SGT-HEM in the range of 5 to 1000 mg/L, and may be extended to higher levels by analysis of a smaller sample volume collected separately.

  19. A novel method of genomic DNA extraction for Cactaceae1

    PubMed Central

    Fehlberg, Shannon D.; Allen, Jessica M.; Church, Kathleen

    2013-01-01

    • Premise of the study: Genetic studies of Cactaceae can at times be impeded by difficult sampling logistics and/or high mucilage content in tissues. Simplifying sampling and DNA isolation through the use of cactus spines has not previously been investigated. • Methods and Results: Several protocols for extracting DNA from spines were tested and modified to maximize yield, amplification, and sequencing. Sampling of and extraction from spines resulted in a simplified protocol overall and complete avoidance of mucilage as compared to typical tissue extractions. Sequences from one nuclear and three plastid regions were obtained across eight genera and 20 species of cacti using DNA extracted from spines. • Conclusions: Genomic DNA useful for amplification and sequencing can be obtained from cactus spines. The protocols described here are valuable for any cactus species, but are particularly useful for investigators interested in sampling living collections, extensive field sampling, and/or conservation genetic studies. PMID:25202521

  20. A comparison of juice extraction methods in the pungency measurement of onion bulbs.

    PubMed

    Yoo, Kil Sun; Lee, Eun Jin; Hamilton, Brian K; Patil, Bhimanagouda S

    2016-02-01

    Onion pungency is estimated by measuring the pyruvic acid content in juice extracted from fresh tissues. We compared pyruvic acid content and its variation in the juices extracted by the pressing, maceration, blending with no water, or blending with water (blend/water) methods. There were considerable differences in the pyruvic acid content and coefficient of variation (CV) among these methods, and there was an interaction between the onion cultivars and the juice extraction methods. The pressing method showed over 30% CV in the quartered or composite samples. The blend/water method showed the greatest pyruvic acid content in the shortday-type ('TG1015Y' and 'Texas Early White') onions, while the pressing method showed the greatest pyruvic acid content in the longday-type onions. The blend/water method, which gave ratios between 1:1 and 1:4 (w/w), showed the same pyruvic acid content. The blending (no water) method had the highest correlation, followed by the maceration method. The lowest correlations were found with the pressing method and the blend/water method. Complete homogenisation of tissues with 1:1 or greater ratios of water was necessary for the maximum consistency and full development of the pyruvic acid reaction for onion pungency measurement. © 2015 Society of Chemical Industry.

  1. Field and laboratory determination of a poly(vinyl/vinylidene chloride) additive in brick mortar.

    PubMed

    Law, S L; Newman, J H; Ptak, F L

    1990-02-01

    A polymerized vinyl/vinylidene chloride additive, used in brick mortar during the 60s and 70s, is detected at the building site by the field method, which employs a commercially available chloride test strip. The field test results can then be verified by the laboratory methods. In one method, total chlorine in the mortar is determined by an oxygen-bomb method and the additive chloride is determined by difference after water-soluble chlorides have been determined on a separate sample. In the second method, the polymerized additive is extracted directly from the mortar with tetrahydrofuran (THF). The difference in weight before and after extraction of the additive gives the weight of additive in the mortar. Evaporation of the THF from the extract leaves a thin film of the polymer, which gives an infrared "fingerprint" spectrum characteristic of the additive polymer.

  2. Comparison and evaluation of volatile oils from three different extraction methods for some Thai fragrant flowers.

    PubMed

    Paibon, W; Yimnoi, C-A; Tembab, N; Boonlue, W; Jampachaisri, K; Nuengchamnong, N; Waranuch, N; Ingkaninan, K

    2011-04-01

    Several tropical flowers have distinctive fragrances which are very appealing to use in perfumery, cosmetics and spa. However, to obtain a 'natural fragrance' from the flower is a challenge as the scent could change during the extraction process. The aim of the study is to find the suitable procedure for extraction of volatile oils from some Thai fragrant flowers. Three different methods: hydrodistillation, solvent extraction and enfleurage methods have been applied for the extraction of volatile oil from Jasminum sambac L. Aiton; Oleaceae (jasmine). The quantities and quality of jasmine volatile oils obtained from the different tested methods were compared. The solvent extraction method using 95% ethanol provided the greatest level of oil yield. However, sensory evaluation using preference test showed that the scents of the volatile oils from solvent extraction using diethyl ether and from enfleurage method were the closest to the fresh flowers compared with the volatile oils obtained from other methods. Their chemical constituents were analysed using gas chromatography coupled with mass spectrometer. Both volatile oils were then evaluated using a triangle discrimination test. From the triangle test, we found that 14 panellists from the total of 36 could not distinguish between the scents of jasmine oil from enfleurage and fresh jasmine flowers whereas only one panellist could not distinguish between the scent of jasmine oil from the solvent extraction and fresh jasmine flowers. These results suggest that the scent of the volatile oil obtained from the enfleurage method was the closest to fresh flowers compared with that obtained from other methods. This method was then successfully applied for extraction of volatile oils from three other Thai fragrant flowers, Michelia alba DC.; Magnoliaceae, Millingtonia hortensis L.; Bignoniaceae and Hedychium coronarium J. Konig; Zingiberaceae. © 2010 The Authors. Journal compilation © 2010 Society of Cosmetic Scientists and

  3. Screening of extraction methods for glycoproteins from jellyfish ( Rhopilema esculentum) oral-arms by high performance liquid chromatography

    NASA Astrophysics Data System (ADS)

    Ren, Guoyan; Li, Bafang; Zhao, Xue; Zhuang, Yongliang; Yan, Mingyan; Hou, Hu; Zhang, Xiukun; Chen, Li

    2009-03-01

    In order to select an optimum extraction method for the target glycoprotein (TGP) from jellyfish ( Rhopilema esculentum) oral-arms, a high performance liquid chromatography (HPLC)-assay for the determination of the TGP was developed. Purified target glycoprotein was taken as a standard glycoprotein. The results showed that the calibration curves for peak area plotted against concentration for TGP were linear ( r = 0.9984, y = 4.5895 x+47.601) over concentrations ranging from 50 to 400 mgL-1. The mean extraction recovery was 97.84% (CV2.60%). The fractions containing TGP were isolated from jellyfish ( R. esculentum) oral-arms by four extraction methods: 1) water extraction (WE), 2) phosphate buffer solution (PBS) extraction (PE), 3) ultrasound-assisted water extraction (UA-WE), 4) ultrasound-assisted PBS extraction (UA-PE). The lyophilized extract was dissolved in Milli-Q water and analyzed directly on a short TSK-GEL G4000PWXL (7.8 mm×300 mm) column. Our results indicated that the UA-PE method was the optimum extraction method selected by HPLC.

  4. A rule-based named-entity recognition method for knowledge extraction of evidence-based dietary recommendations

    PubMed Central

    2017-01-01

    Evidence-based dietary information represented as unstructured text is a crucial information that needs to be accessed in order to help dietitians follow the new knowledge arrives daily with newly published scientific reports. Different named-entity recognition (NER) methods have been introduced previously to extract useful information from the biomedical literature. They are focused on, for example extracting gene mentions, proteins mentions, relationships between genes and proteins, chemical concepts and relationships between drugs and diseases. In this paper, we present a novel NER method, called drNER, for knowledge extraction of evidence-based dietary information. To the best of our knowledge this is the first attempt at extracting dietary concepts. DrNER is a rule-based NER that consists of two phases. The first one involves the detection and determination of the entities mention, and the second one involves the selection and extraction of the entities. We evaluate the method by using text corpora from heterogeneous sources, including text from several scientifically validated web sites and text from scientific publications. Evaluation of the method showed that drNER gives good results and can be used for knowledge extraction of evidence-based dietary recommendations. PMID:28644863

  5. An efficient method for extraction of astaxanthin from green alga Haematococcus pluvialis.

    PubMed

    Sarada, R; Vidhyavathi, R; Usha, D; Ravishankar, G A

    2006-10-04

    Haematococcus pluvialis is one of the potent organisms for production of astaxanthin, a high value ketocarotenoid. Astaxanthin is accumulated in thick-walled cyst cells of Haematococcus. The thick cell wall is made up of sporopollenin-like material, algaenan, which hinders solvent extraction of astaxanthin. In the present study, an improved method for extraction of astaxanthin without homogenization of cells is reported. Extractability of astaxanthin from cyst cells was evaluated by treating cells with various solvents and pretreating the cells with organic and mineral acids at 70 degrees C followed by acetone extraction. Hydrochloric acid treatment facilitated 86-94% extractability of astaxanthin. Treatment time, temperature, and concentration of the acid were found to be critical factors for maximum extractability. The treatment did not affect the astaxanthin ester profile and the treated cells can be preserved until further use.

  6. Pressure-Assisted Chelating Extraction as a Teaching Tool in Instrumental Analysis

    ERIC Educational Resources Information Center

    Sadik, Omowunmi A.; Wanekaya, Adam K.; Yevgeny, Gelfand

    2004-01-01

    A novel instrumental-digestion technique using pressure-assisted chelating extraction (PACE), for undergraduate laboratory is reported. This procedure is used for exposing students to safe sample-preparation techniques, for correlating wet-chemical methods with modern instrumental analysis and comparing the performance of PACE with conventional…

  7. Extraction of polycyclic aromatic hydrocarbons and organochlorine pesticides from soils: a comparison between Soxhlet extraction, microwave-assisted extraction and accelerated solvent extraction techniques.

    PubMed

    Wang, Wentao; Meng, Bingjun; Lu, Xiaoxia; Liu, Yu; Tao, Shu

    2007-10-29

    The methods of simultaneous extraction of polycyclic aromatic hydrocarbons (PAHs) and organochlorine pesticides (OCPs) from soils using Soxhlet extraction, microwave-assisted extraction (MAE) and accelerated solvent extraction (ASE) were established, and the extraction efficiencies using the three methods were systemically compared from procedural blank, limits of detection and quantification, method recovery and reproducibility, method chromatogram and other factors. In addition, soils with different total organic carbon contents were used to test the extraction efficiencies of the three methods. The results showed that the values obtained in this study were comparable with the values reported by other studies. In some respects such as method recovery and reproducibility, there were no significant differences among the three methods for the extraction of PAHs and OCPs. In some respects such as procedural blank and limits of detection and quantification, there were significant differences among the three methods. Overall, ASE had the best extraction efficiency compared to MAE and Soxhlet extraction, and the extraction efficiencies of MAE and Soxhlet extraction were comparable to each other depending on the property such as TOC content of the studied soil. Considering other factors such as solvent consumption and extraction time, ASE and MAE are preferable to Soxhlet extraction.

  8. High Throughput Method of Extracting and Counting Strontium-90 in Urine

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

    Shkrob, I.; Kaminski, M.; Mertz, C.

    2016-03-01

    A method has been developed for the rapid extraction of Sr-90 from the urine of individuals exposed to radiation in a terrorist attack. The method employs two chromatographic ion-exchange materials: Diphonix resin and Sr resin, both of which are commercially available. The Diphonix resin reduces the alkali ion concentrations below 10 mM, and the Sr resin concentrates and decontaminates strontium-90. Experimental and calculational data are given for a variety of test conditions. On the basis of these results, a flowsheet has been developed for the rapid concentration and extraction of Sr-90 from human urine samples for subsequent beta-counting.

  9. A new method for fast extraction and determination of chlorophylls in natural water.

    PubMed

    Qiu, Nianwei; Wang, Xiushun; Zhou, Feng

    2018-01-26

    Algae collection and chlorophyll extraction are two troublesome steps in the traditional methods used for the determination of chlorophyll concentration in natural water. A new method was established in this study for fast collection and extraction of chlorophyll. Based on our results, the optimum centrifugation condition for collecting algae was determined as: 5000 g for 15 min at 4 °C, and the optimum dilution ratio of dimethyl sulfoxide: 90% acetone was 1:4. The specific steps were as follows: the algae in water samples were collected by centrifugation at 5000 g at 4 °C for 15 min. The precipitated algae were suspended with 2 mL DMSO. Then the sample was transferred to a 15 mL centrifuge tube, and the tube was incubated at 65 °C for 1-2 h in the dark until the sample turned white. After cooling, the chlorophyll extract was diluted with 8 mL 90% acetone, before centrifugation at 5000 g for 5 min. The absorbance values of the supernatants at 750, 664, 647 and 630 nm were used for the calculation of chlorophyll concentrations by the trichromatic equations. This new method saved the filter cost, simplified the extraction process, improved the algae acquisition efficiency, and accelerated the chlorophyll extraction rate.

  10. Combining Feature Extraction Methods to Assist the Diagnosis of Alzheimer's Disease.

    PubMed

    Segovia, F; Górriz, J M; Ramírez, J; Phillips, C

    2016-01-01

    Neuroimaging data as (18)F-FDG PET is widely used to assist the diagnosis of Alzheimer's disease (AD). Looking for regions with hypoperfusion/ hypometabolism, clinicians may predict or corroborate the diagnosis of the patients. Modern computer aided diagnosis (CAD) systems based on the statistical analysis of whole neuroimages are more accurate than classical systems based on quantifying the uptake of some predefined regions of interests (ROIs). In addition, these new systems allow determining new ROIs and take advantage of the huge amount of information comprised in neuroimaging data. A major branch of modern CAD systems for AD is based on multivariate techniques, which analyse a neuroimage as a whole, considering not only the voxel intensities but also the relations among them. In order to deal with the vast dimensionality of the data, a number of feature extraction methods have been successfully applied. In this work, we propose a CAD system based on the combination of several feature extraction techniques. First, some commonly used feature extraction methods based on the analysis of the variance (as principal component analysis), on the factorization of the data (as non-negative matrix factorization) and on classical magnitudes (as Haralick features) were simultaneously applied to the original data. These feature sets were then combined by means of two different combination approaches: i) using a single classifier and a multiple kernel learning approach and ii) using an ensemble of classifier and selecting the final decision by majority voting. The proposed approach was evaluated using a labelled neuroimaging database along with a cross validation scheme. As conclusion, the proposed CAD system performed better than approaches using only one feature extraction technique. We also provide a fair comparison (using the same database) of the selected feature extraction methods.

  11. Analysis of Lard in Lipstick Formulation Using FTIR Spectroscopy and Multivariate Calibration: A Comparison of Three Extraction Methods.

    PubMed

    Waskitho, Dri; Lukitaningsih, Endang; Sudjadi; Rohman, Abdul

    2016-01-01

    Analysis of lard extracted from lipstick formulation containing castor oil has been performed using FTIR spectroscopic method combined with multivariate calibration. Three different extraction methods were compared, namely saponification method followed by liquid/liquid extraction with hexane/dichlorometane/ethanol/water, saponification method followed by liquid/liquid extraction with dichloromethane/ethanol/water, and Bligh & Dyer method using chloroform/methanol/water as extracting solvent. Qualitative and quantitative analysis of lard were performed using principle component (PCA) and partial least square (PLS) analysis, respectively. The results showed that, in all samples prepared by the three extraction methods, PCA was capable of identifying lard at wavelength region of 1200-800 cm -1 with the best result was obtained by Bligh & Dyer method. Furthermore, PLS analysis at the same wavelength region used for qualification showed that Bligh and Dyer was the most suitable extraction method with the highest determination coefficient (R 2 ) and the lowest root mean square error of calibration (RMSEC) as well as root mean square error of prediction (RMSEP) values.

  12. Comparison of Different Protein Extraction Methods for Gel-Based Proteomic Analysis of Ganoderma spp.

    PubMed

    Al-Obaidi, Jameel R; Saidi, Noor Baity; Usuldin, Siti Rokhiyah Ahmad; Hussin, Siti Nahdatul Isnaini Said; Yusoff, Noornabeela Md; Idris, Abu Seman

    2016-04-01

    Ganoderma species are a group of fungi that have the ability to degrade lignin polymers and cause severe diseases such as stem and root rot and can infect economically important plants and perennial crops such as oil palm, especially in tropical countries such as Malaysia. Unfortunately, very little is known about the complex interplay between oil palm and Ganoderma in the pathogenesis of the diseases. Proteomic technologies are simple yet powerful tools in comparing protein profile and have been widely used to study plant-fungus interaction. A critical step to perform a good proteome research is to establish a method that gives the best quality and a wide coverage of total proteins. Despite the availability of various protein extraction protocols from pathogenic fungi in the literature, no single extraction method was found suitable for all types of pathogenic fungi. To develop an optimized protein extraction protocol for 2-DE gel analysis of Ganoderma spp., three previously reported protein extraction protocols were compared: trichloroacetic acid, sucrose and phenol/ammonium acetate in methanol. The third method was found to give the most reproducible gels and highest protein concentration. Using the later method, a total of 10 protein spots (5 from each species) were successfully identified. Hence, the results from this study propose phenol/ammonium acetate in methanol as the most effective protein extraction method for 2-DE proteomic studies of Ganoderma spp.

  13. A simple method for the small scale synthesis and solid-phase extraction purification of steroid sulfates.

    PubMed

    Waller, Christopher C; McLeod, Malcolm D

    2014-12-01

    Steroid sulfates are a major class of steroid metabolite that are of growing importance in fields such as anti-doping analysis, the detection of residues in agricultural produce or medicine. Despite this, many steroid sulfate reference materials may have limited or no availability hampering the development of analytical methods. We report simple protocols for the rapid synthesis and purification of steroid sulfates that are suitable for adoption by analytical laboratories. Central to this approach is the use of solid-phase extraction (SPE) for purification, a technique routinely used for sample preparation in analytical laboratories around the world. The sulfate conjugates of sixteen steroid compounds encompassing a wide range of steroid substitution patterns and configurations are prepared, including the previously unreported sulfate conjugates of the designer steroids furazadrol (17β-hydroxyandrostan[2,3-d]isoxazole), isofurazadrol (17β-hydroxyandrostan[3,2-c]isoxazole) and trenazone (17β-hydroxyestra-4,9-dien-3-one). Structural characterization data, together with NMR and mass spectra are reported for all steroid sulfates, often for the first time. The scope of this approach for small scale synthesis is highlighted by the sulfation of 1μg of testosterone (17β-hydroxyandrost-4-en-3-one) as monitored by liquid chromatography-mass spectrometry (LCMS). Copyright © 2014 Elsevier Inc. All rights reserved.

  14. PPI-IRO: a two-stage method for protein-protein interaction extraction based on interaction relation ontology.

    PubMed

    Li, Chuan-Xi; Chen, Peng; Wang, Ru-Jing; Wang, Xiu-Jie; Su, Ya-Ru; Li, Jinyan

    2014-01-01

    Mining Protein-Protein Interactions (PPIs) from the fast-growing biomedical literature resources has been proven as an effective approach for the identification of biological regulatory networks. This paper presents a novel method based on the idea of Interaction Relation Ontology (IRO), which specifies and organises words of various proteins interaction relationships. Our method is a two-stage PPI extraction method. At first, IRO is applied in a binary classifier to determine whether sentences contain a relation or not. Then, IRO is taken to guide PPI extraction by building sentence dependency parse tree. Comprehensive and quantitative evaluations and detailed analyses are used to demonstrate the significant performance of IRO on relation sentences classification and PPI extraction. Our PPI extraction method yielded a recall of around 80% and 90% and an F1 of around 54% and 66% on corpora of AIMed and BioInfer, respectively, which are superior to most existing extraction methods.

  15. Optimization of cloud point extraction and solid phase extraction methods for speciation of arsenic in natural water using multivariate technique.

    PubMed

    Baig, Jameel A; Kazi, Tasneem G; Shah, Abdul Q; Arain, Mohammad B; Afridi, Hassan I; Kandhro, Ghulam A; Khan, Sumaira

    2009-09-28

    The simple and rapid pre-concentration techniques viz. cloud point extraction (CPE) and solid phase extraction (SPE) were applied for the determination of As(3+) and total inorganic arsenic (iAs) in surface and ground water samples. The As(3+) was formed complex with ammonium pyrrolidinedithiocarbamate (APDC) and extracted by surfactant-rich phases in the non-ionic surfactant Triton X-114, after centrifugation the surfactant-rich phase was diluted with 0.1 mol L(-1) HNO(3) in methanol. While total iAs in water samples was adsorbed on titanium dioxide (TiO(2)); after centrifugation, the solid phase was prepared to be slurry for determination. The extracted As species were determined by electrothermal atomic absorption spectrometry. The multivariate strategy was applied to estimate the optimum values of experimental factors for the recovery of As(3+) and total iAs by CPE and SPE. The standard addition method was used to validate the optimized methods. The obtained result showed sufficient recoveries for As(3+) and iAs (>98.0%). The concentration factor in both cases was found to be 40.

  16. Extraction of the ANC from the 10Be(d, p)11Be transfer reaction using the ADWA method

    NASA Astrophysics Data System (ADS)

    Yang, J.; Capel, P.

    2018-05-01

    A halo nucleus is built from a core and at least one weakly bound neutron or proton. To understand this unique cluster structure, lots of efforts have been undertaken. During the past decades, the (d, p) reaction has been widely used in experiments and has become an important tool for extracting single-particle properties of nuclei. In this work, our goal is to obtain the Asymptotic Normalization Coefficient (ANC) of the halo nuclei 11Be using the ADWA method. We perform the analysis for the 10Be(d, p)11Be stripping reaction at Ed =21.4, 18, 15, and 12MeV for the ground state and first excited state of the composite nucleus 11Be. The experimental measurement was carried out at Oak Ridge National Laboratory by Schmitt et al. [1] The sensitivity of the calculations to the optical potential choice is also checked. Overall, the transfer process becomes more peripheral at lower energies and forward angles. Investigation in this area is the best way to extract a reliable ANC from the experimental data. For the ground state of 11Be, the ANC obtained using our method (C={0.785}-0.030+0.026{\\text{fm}}-1/2) shows perfect agreement with the one obtained by Ab initio calculations (C Ab=0.786 fm‑1/2) [2].

  17. Extraction of basil leaves (ocimum canum) oleoresin with ethyl acetate solvent by using soxhletation method

    NASA Astrophysics Data System (ADS)

    Tambun, R.; Purba, R. R. H.; Ginting, H. K.

    2017-09-01

    The goal of this research is to produce oleoresin from basil leaves (Ocimum canum) by using soxhletation method and ethyl acetate as solvent. Basil commonly used in culinary as fresh vegetables. Basil contains essential oils and oleoresin that are used as flavouring agent in food, in cosmetic and ingredient in traditional medicine. The extraction method commonly used to obtain oleoresin is maceration. The problem of this method is many solvents necessary and need time to extract the raw material. To resolve the problem and to produce more oleoresin, we use soxhletation method with a combination of extraction time and ratio from the material with a solvent. The analysis consists of yield, density, refractive index, and essential oil content. The best treatment of basil leaves oleoresin extraction is at ratio of material and solvent 1:6 (w / v) for 6 hours extraction time. In this condition, the yield of basil oleoresin is 20.152%, 0.9688 g/cm3 of density, 1.502 of refractive index, 15.77% of essential oil content, and the colour of oleoresin product is dark-green.

  18. Alternative oil extraction methods from Echium plantagineum L. seeds using advanced techniques and green solvents.

    PubMed

    Castejón, Natalia; Luna, Pilar; Señoráns, Francisco J

    2018-04-01

    The edible oil processing industry involves large losses of organic solvent into the atmosphere and long extraction times. In this work, fast and environmentally friendly alternatives for the production of echium oil using green solvents are proposed. Advanced extraction techniques such as Pressurized Liquid Extraction (PLE), Microwave Assisted Extraction (MAE) and Ultrasound Assisted Extraction (UAE) were evaluated to efficiently extract omega-3 rich oil from Echium plantagineum seeds. Extractions were performed with ethyl acetate, ethanol, water and ethanol:water to develop a hexane-free processing method. Optimal PLE conditions with ethanol at 150 °C during 10 min produced a very similar oil yield (31.2%) to Soxhlet using hexane for 8 h (31.3%). UAE optimized method with ethanol at mild conditions (55 °C) produced a high oil yield (29.1%). Consequently, advanced extraction techniques showed good lipid yields and furthermore, the produced echium oil had the same omega-3 fatty acid composition than traditionally extracted oil. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Method 1664: N-hexane extractable material (hem) and silica gel treated n-hexane extractable material (SGT-HEM) by extraction and gravimetry (oil and grease and total petroleum hydrocarbons), April 1995

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

    NONE

    Method 1664 was developed by the United States Environmental Protection Agency Office of Science and Technology to replace previously used gravimetric procedures that employed Freon-113, a Class I CFC, as the extraction solvent for the determination of oil and grease and petroleum hydrocarbons. Method 1664 is a performance-based method applicable to aqueous matrices that requires the use of n-hexane as the extraction solvent and gravimetry as the determinative technique. In addition, QC procedures designed to monitor precision and accuracy have been incorporated into Method 1664.

  20. Extraction and Determination of Cyproheptadine in Human Urine by DLLME-HPLC Method

    PubMed Central

    Maham, Mehdi; Kiarostami, Vahid; Waqif-Husain, Syed; Abroomand-Azar, Parviz; Tehrani, Mohammad Saber; Khoeini Sharifabadi, Malihe; Afrouzi, Hossein; Shapouri, MahmoudReza; Karami-Osboo, Rouhollah

    2013-01-01

    Novel dispersive liquid-liquid microextraction (DLLME), coupled with high performance liquid chromatography with photodiode array detection (HPLC-DAD) has been applied for the extraction and determination of cyproheptadine (CPH), an antihistamine, in human urine samples. In this method, 0.6 mL of acetonitrile (disperser solvent) containing 30 μL of carbon tetrachloride (extraction solvent) was rapidly injected by a syringe into 5 mL urine sample. After centrifugation, the sedimented phase containing enriched analyte was dissolved in acetonitrile and an aliquot of this solution injected into the HPLC system for analysis. Development of DLLME procedure includes optimization of some important parameters such as kind and volume of extraction and disperser solvent, pH and salt addition. The proposed method has good linearity in the range of 0.02-4.5 μg mL-1 and low detection limit (13.1 ng mL-1). The repeatability of the method, expressed as relative standard deviation was 4.9% (n = 3). This method has also been applied to the analysis of real urine samples with satisfactory relative recoveries in the range of 91.6-101.0%. PMID:24250605

  1. Extraction and Determination of Cyproheptadine in Human Urine by DLLME-HPLC Method.

    PubMed

    Maham, Mehdi; Kiarostami, Vahid; Waqif-Husain, Syed; Abroomand-Azar, Parviz; Tehrani, Mohammad Saber; Khoeini Sharifabadi, Malihe; Afrouzi, Hossein; Shapouri, Mahmoudreza; Karami-Osboo, Rouhollah

    2013-01-01

    Novel dispersive liquid-liquid microextraction (DLLME), coupled with high performance liquid chromatography with photodiode array detection (HPLC-DAD) has been applied for the extraction and determination of cyproheptadine (CPH), an antihistamine, in human urine samples. In this method, 0.6 mL of acetonitrile (disperser solvent) containing 30 μL of carbon tetrachloride (extraction solvent) was rapidly injected by a syringe into 5 mL urine sample. After centrifugation, the sedimented phase containing enriched analyte was dissolved in acetonitrile and an aliquot of this solution injected into the HPLC system for analysis. Development of DLLME procedure includes optimization of some important parameters such as kind and volume of extraction and disperser solvent, pH and salt addition. The proposed method has good linearity in the range of 0.02-4.5 μg mL(-1) and low detection limit (13.1 ng mL(-1)). The repeatability of the method, expressed as relative standard deviation was 4.9% (n = 3). This method has also been applied to the analysis of real urine samples with satisfactory relative recoveries in the range of 91.6-101.0%.

  2. Validation for Vegetation Green-up Date Extracted from GIMMS NDVI and NDVI3g Using Variety of Methods

    NASA Astrophysics Data System (ADS)

    Chang, Q.; Jiao, W.

    2017-12-01

    Phenology is a sensitive and critical feature of vegetation change that has regarded as a good indicator in climate change studies. So far, variety of remote sensing data sources and phenology extraction methods from satellite datasets have been developed to study the spatial-temporal dynamics of vegetation phenology. However, the differences between vegetation phenology results caused by the varies satellite datasets and phenology extraction methods are not clear, and the reliability for different phenology results extracted from remote sensing datasets is not verified and compared using the ground observation data. Based on three most popular remote sensing phenology extraction methods, this research calculated the Start of the growing season (SOS) for each pixels in the Northern Hemisphere for two kinds of long time series satellite datasets: GIMMS NDVIg (SOSg) and GIMMS NDVI3g (SOS3g). The three methods used in this research are: maximum increase method, dynamic threshold method and midpoint method. Then, this study used SOS calculated from NEE datasets (SOS_NEE) monitored by 48 eddy flux tower sites in global flux website to validate the reliability of six phenology results calculated from remote sensing datasets. Results showed that both SOSg and SOS3g extracted by maximum increase method are not correlated with ground observed phenology metrics. SOSg and SOS3g extracted by the dynamic threshold method and midpoint method are both correlated with SOS_NEE significantly. Compared with SOSg extracted by the dynamic threshold method, SOSg extracted by the midpoint method have a stronger correlation with SOS_NEE. And, the same to SOS3g. Additionally, SOSg showed stronger correlation with SOS_NEE than SOS3g extracted by the same method. SOS extracted by the midpoint method from GIMMS NDVIg datasets seemed to be the most reliable results when validated with SOS_NEE. These results can be used as reference for data and method selection in future's phenology study.

  3. Solvent Extraction of Copper: An Extractive Metallurgy Exercise for Undergraduate Teaching Laboratories

    ERIC Educational Resources Information Center

    Smellie, Iain A.; Forgan, Ross S.; Brodie, Claire; Gavine, Jack S.; Harris, Leanne; Houston, Daniel; Hoyland, Andrew D.; McCaughan, Rory P.; Miller, Andrew J.; Wilson, Liam; Woodhall, Fiona M.

    2016-01-01

    A multidisciplinary experiment for advanced undergraduate students has been developed in the context of extractive metallurgy. The experiment serves as a model of an important modern industrial process that combines aspects of organic/inorganic synthesis and analysis. Students are tasked to prepare a salicylaldoxime ligand and samples of the…

  4. A method for quickly and exactly extracting hepatic vein

    NASA Astrophysics Data System (ADS)

    Xiong, Qing; Yuan, Rong; Wang, Luyao; Wang, Yanchun; Li, Zhen; Hu, Daoyu; Xie, Qingguo

    2013-02-01

    It is of vital importance that providing detailed and accurate information about hepatic vein (HV) for liver surgery planning, such as pre-operative planning of living donor liver transplantation (LDLT). Due to the different blood flow rate of intra-hepatic vascular systems and the restrictions of CT scan, it is common that HV and hepatic portal vein (HPV) are both filled with contrast medium during the scan and in high intensity in the hepatic venous phase images. As a result, the HV segmentation result obtained from the hepatic venous phase images is always contaminated by HPV which makes accurate HV modeling difficult. In this paper, we proposed a method for quick and accurate HV extraction. Based on the topological structure of intra-hepatic vessels, we analyzed the anatomical features of HV and HPV. According to the analysis, three conditions were presented to identify the nodes that connect HV with HPV in the topological structure, and thus to distinguish HV from HPV. The method costs less than one minute to extract HV and provides a correct and detailed HV model even with variations in vessels. Evaluated by two experienced radiologists, the accuracy of the HV model obtained from our method is over 97%. In the following work, we will extend our work to a comprehensive clinical evaluation and apply this method to actual LDLT surgical planning.

  5. Evaluating Point of Sale Tobacco Marketing Using Behavioral Laboratory Methods

    PubMed Central

    Robinson, Jason D.; Drobes, David J.; Brandon, Thomas H.; Wetter, David W.; Cinciripini, Paul M.

    2018-01-01

    With passage of the 2009 Family Smoking Prevention and Tobacco Control Act, the FDA has authority to regulate tobacco advertising. As bans on traditional advertising venues and promotion of tobacco products have grown, a greater emphasis has been placed on brand exposure and price promotion in displays of products at the point-of-sale (POS). POS marketing seeks to influence attitudes and behavior towards tobacco products using a variety of explicit and implicit messaging approaches. Behavioral laboratory methods have the potential to provide the FDA with a strong scientific base for regulatory actions and a model for testing future manipulations of POS advertisements. We review aspects of POS marketing that potentially influence smoking behavior, including branding, price promotions, health claims, the marketing of emerging tobacco products, and tobacco counter-advertising. We conceptualize how POS marketing potentially influence individual attention, memory, implicit attitudes, and smoking behavior. Finally, we describe specific behavioral laboratory methods that can be adapted to measure the impact of POS marketing on these domains.

  6. Extraction optimization and UHPLC method development for determination of the 20-hydroxyecdysone in Sida tuberculata leaves.

    PubMed

    da Rosa, Hemerson S; Koetz, Mariana; Santos, Marí Castro; Jandrey, Elisa Helena Farias; Folmer, Vanderlei; Henriques, Amélia Teresinha; Mendez, Andreas Sebastian Loureiro

    2018-04-01

    Sida tuberculata (ST) is a Malvaceae species widely distributed in Southern Brazil. In traditional medicine, ST has been employed as hypoglycemic, hypocholesterolemic, anti-inflammatory and antimicrobial. Additionally, this species is chemically characterized by flavonoids, alkaloids and phytoecdysteroids mainly. The present work aimed to optimize the extractive technique and to validate an UHPLC method for the determination of 20-hydroxyecdsone (20HE) in the ST leaves. Box-Behnken Design (BBD) was used in method optimization. The extractive methods tested were: static and dynamic maceration, ultrasound, ultra-turrax and reflux. In the Box-Behnken three parameters were evaluated in three levels (-1, 0, +1), particle size, time and plant:solvent ratio. In validation method, the parameters of selectivity, specificity, linearity, limits of detection and quantification (LOD, LOQ), precision, accuracy and robustness were evaluated. The results indicate static maceration as better technique to obtain 20HE peak area in ST extract. The optimal extraction from surface response methodology was achieved with the parameters granulometry of 710 nm, 9 days of maceration and plant:solvent ratio 1:54 (w/v). The UHPLC-PDA analytical developed method showed full viability of performance, proving to be selective, linear, precise, accurate and robust for 20HE detection in ST leaves. The average content of 20HE was 0.56% per dry extract. Thus, the optimization of extractive method in ST leaves increased the concentration of 20HE in crude extract, and a reliable method was successfully developed according to validation requirements and in agreement with current legislation. Copyright © 2018 Elsevier Inc. All rights reserved.

  7. Comparison of on-site field measured inorganic arsenic in rice with laboratory measurements using a field deployable method: Method validation.

    PubMed

    Mlangeni, Angstone Thembachako; Vecchi, Valeria; Norton, Gareth J; Raab, Andrea; Krupp, Eva M; Feldmann, Joerg

    2018-10-15

    A commercial arsenic field kit designed to measure inorganic arsenic (iAs) in water was modified into a field deployable method (FDM) to measure iAs in rice. While the method has been validated to give precise and accurate results in the laboratory, its on-site field performance has not been evaluated. This study was designed to test the method on-site in Malawi in order to evaluate its accuracy and precision in determination of iAs on-site by comparing with a validated reference method and giving original data on inorganic arsenic in Malawian rice and rice-based products. The method was validated by using the established laboratory-based HPLC-ICPMS. Statistical tests indicated there were no significant differences between on-site and laboratory iAs measurements determined using the FDM (p = 0.263, ά = 0.05) and between on-site measurements and measurements determined using HPLC-ICP-MS (p = 0.299, ά = 0.05). This method allows quick (within 1 h) and efficient screening of rice containing iAs concentrations on-site. Copyright © 2018 Elsevier Ltd. All rights reserved.

  8. New Laboratory Methods for Characterizing the Immersion Factors for Irradiance

    NASA Technical Reports Server (NTRS)

    Hooker, Stanford B. (Editor); Firestone, Elaine R. (Editor); Zibordi, Giuseppe; DAlimonte, Davide; vaderLinde, Dirk; Brown, James W.

    2003-01-01

    The experimental determination of the immersion factor, I(sub f)(lambda), of irradiance collectors is a requirement of any in-water radiometer. The eighth SeaWiFS Intercalibration Round-Robin Experiment (SIRREX-8) showed different implementations, at different laboratories, of the same I(sub f)(lambda) measurement protocol. The different implementations make use of different setups, volumes, and water types. Consequently, they exhibit different accuracies and require different execution times for characterizing an irradiance sensor. In view of standardizing the characterization of I(sub f)(lambda) values for in-water radiometers, together with an increase in the accuracy of methods and a decrease in the execution time, alternative methods are presented, and assessed versus the traditional method. The proposed new laboratory methods include: a) the continuous method, in which optical measurements taken with discrete water depths are substituted by continuous profiles created by removing the water from the water vessel at a constant flow rate (which significantly reduces the time required for the characterization of a single radiometer); and b) the Compact Portable Advanced Characterization Tank (ComPACT) method, in which the commonly used large tanks are replaced by a small water vessel, thereby allowing the determination of I(sub f)(lambda) values with a small water volume, and more importantly, permitting I(sub f)(lambda) characterizations with pure water. Intercomparisons between the continuous and the traditional method showed results within the variance of I(sub f) (lambda) determinations. The use of the continuous method, however, showed a much shorter realization time. Intercomparisons between the ComPACT and the traditional method showed generally higher I(sub f)(lambda) values for the former. This is in agreement with the generalized expectations of a reduction in scattering effects, because of the use of pure water with the ComPACT method versus the use of

  9. Effects of DNA extraction and purification methods on real-time quantitative PCR analysis of Roundup Ready soybean.

    PubMed

    Demeke, Tigst; Ratnayaka, Indira; Phan, Anh

    2009-01-01

    The quality of DNA affects the accuracy and repeatability of quantitative PCR results. Different DNA extraction and purification methods were compared for quantification of Roundup Ready (RR) soybean (event 40-3-2) by real-time PCR. DNA was extracted using cetylmethylammonium bromide (CTAB), DNeasy Plant Mini Kit, and Wizard Magnetic DNA purification system for food. CTAB-extracted DNA was also purified using the Zymo (DNA Clean & Concentrator 25 kit), Qtip 100 (Qiagen Genomic-Tip 100/G), and QIAEX II Gel Extraction Kit. The CTAB extraction method provided the largest amount of DNA, and the Zymo purification kit resulted in the highest percentage of DNA recovery. The Abs260/280 and Abs260/230 ratios were less than the expected values for some of the DNA extraction and purification methods used, indicating the presence of substances that could inhibit PCR reactions. Real-time quantitative PCR results were affected by the DNA extraction and purification methods used. Further purification or dilution of the CTAB DNA was required for successful quantification of RR soybean. Less variability of quantitative PCR results was observed among experiments and replications for DNA extracted and/or purified by CTAB, CTAB+Zymo, CTAB+Qtip 100, and DNeasy methods. Correct and repeatable results for real-time PCR quantification of RR soybean were achieved using CTAB DNA purified with Zymo and Qtip 100 methods.

  10. Validation of a Laboratory Method for Evaluating Dynamic Properties of Reconstructed Equine Racetrack Surfaces

    PubMed Central

    Setterbo, Jacob J.; Chau, Anh; Fyhrie, Patricia B.; Hubbard, Mont; Upadhyaya, Shrini K.; Symons, Jennifer E.; Stover, Susan M.

    2012-01-01

    Background Racetrack surface is a risk factor for racehorse injuries and fatalities. Current research indicates that race surface mechanical properties may be influenced by material composition, moisture content, temperature, and maintenance. Race surface mechanical testing in a controlled laboratory setting would allow for objective evaluation of dynamic properties of surface and factors that affect surface behavior. Objective To develop a method for reconstruction of race surfaces in the laboratory and validate the method by comparison with racetrack measurements of dynamic surface properties. Methods Track-testing device (TTD) impact tests were conducted to simulate equine hoof impact on dirt and synthetic race surfaces; tests were performed both in situ (racetrack) and using laboratory reconstructions of harvested surface materials. Clegg Hammer in situ measurements were used to guide surface reconstruction in the laboratory. Dynamic surface properties were compared between in situ and laboratory settings. Relationships between racetrack TTD and Clegg Hammer measurements were analyzed using stepwise multiple linear regression. Results Most dynamic surface property setting differences (racetrack-laboratory) were small relative to surface material type differences (dirt-synthetic). Clegg Hammer measurements were more strongly correlated with TTD measurements on the synthetic surface than the dirt surface. On the dirt surface, Clegg Hammer decelerations were negatively correlated with TTD forces. Conclusions Laboratory reconstruction of racetrack surfaces guided by Clegg Hammer measurements yielded TTD impact measurements similar to in situ values. The negative correlation between TTD and Clegg Hammer measurements confirms the importance of instrument mass when drawing conclusions from testing results. Lighter impact devices may be less appropriate for assessing dynamic surface properties compared to testing equipment designed to simulate hoof impact (TTD

  11. Optimizing techniques to capture and extract environmental DNA for detection and quantification of fish.

    PubMed

    Eichmiller, Jessica J; Miller, Loren M; Sorensen, Peter W

    2016-01-01

    Few studies have examined capture and extraction methods for environmental DNA (eDNA) to identify techniques optimal for detection and quantification. In this study, precipitation, centrifugation and filtration eDNA capture methods and six commercially available DNA extraction kits were evaluated for their ability to detect and quantify common carp (Cyprinus carpio) mitochondrial DNA using quantitative PCR in a series of laboratory experiments. Filtration methods yielded the most carp eDNA, and a glass fibre (GF) filter performed better than a similar pore size polycarbonate (PC) filter. Smaller pore sized filters had higher regression slopes of biomass to eDNA, indicating that they were potentially more sensitive to changes in biomass. Comparison of DNA extraction kits showed that the MP Biomedicals FastDNA SPIN Kit yielded the most carp eDNA and was the most sensitive for detection purposes, despite minor inhibition. The MoBio PowerSoil DNA Isolation Kit had the lowest coefficient of variation in extraction efficiency between lake and well water and had no detectable inhibition, making it most suitable for comparisons across aquatic environments. Of the methods tested, we recommend using a 1.5 μm GF filter, followed by extraction with the MP Biomedicals FastDNA SPIN Kit for detection. For quantification of eDNA, filtration through a 0.2-0.6 μm pore size PC filter, followed by extraction with MoBio PowerSoil DNA Isolation Kit was optimal. These results are broadly applicable for laboratory studies on carps and potentially other cyprinids. The recommendations can also be used to inform choice of methodology for field studies. © 2015 John Wiley & Sons Ltd.

  12. A method for addressing differences in concentrations of fipronil and three degradates obtained by two different laboratory methods

    USGS Publications Warehouse

    Crawford, Charles G.; Martin, Jeffrey D.

    2017-07-21

    In October 2012, the U.S. Geological Survey (USGS) began measuring the concentration of the pesticide fipronil and three of its degradates (desulfinylfipronil, fipronil sulfide, and fipronil sulfone) by a new laboratory method using direct aqueous-injection liquid chromatography tandem mass spectrometry (DAI LC–MS/MS). This method replaced the previous method—in use since 2002—that used gas chromatography/mass spectrometry (GC/MS). The performance of the two methods is not comparable for fipronil and the three degradates. Concentrations of these four chemical compounds determined by the DAI LC–MS/MS method are substantially lower than the GC/MS method. A method was developed to correct for the difference in concentrations obtained by the two laboratory methods based on a methods comparison field study done in 2012. Environmental and field matrix spike samples to be analyzed by both methods from 48 stream sites from across the United States were sampled approximately three times each for this study. These data were used to develop a relation between the two laboratory methods for each compound using regression analysis. The relations were used to calibrate data obtained by the older method to the new method in order to remove any biases attributable to differences in the methods. The coefficients of the equations obtained from the regressions were used to calibrate over 16,600 observations of fipronil, as well as the three degradates determined by the GC/MS method retrieved from the USGS National Water Information System. The calibrated values were then compared to over 7,800 observations of fipronil and to the three degradates determined by the DAI LC–MS/MS method also retrieved from the National Water Information System. The original and calibrated values from the GC/MS method, along with measures of uncertainty in the calibrated values and the original values from the DAI LC–MS/MS method, are provided in an accompanying data release.

  13. Rapid Solid-Liquid Dynamic Extraction (RSLDE): a New Rapid and Greener Method for Extracting Two Steviol Glycosides (Stevioside and Rebaudioside A) from Stevia Leaves.

    PubMed

    Gallo, Monica; Vitulano, Manuela; Andolfi, Anna; DellaGreca, Marina; Conte, Esterina; Ciaravolo, Martina; Naviglio, Daniele

    2017-06-01

    Stevioside and rebaudioside A are the main diterpene glycosides present in the leaves of the Stevia rebaudiana plant, which is used in the production of foods and low-calorie beverages. The difficulties associated with their extraction and purification are currently a problem for the food processing industries. The objective of this study was to develop an effective and economically viable method to obtain a high-quality product while trying to overcome the disadvantages derived from the conventional transformation processes. For this reason, extractions were carried out using a conventional maceration (CM) and a cyclically pressurized extraction known as rapid solid-liquid dynamic extraction (RSLDE) by the Naviglio extractor (NE). After only 20 min of extraction using the NE, a quantity of rebaudioside A and stevioside equal to 1197.8 and 413.6 mg/L was obtained, respectively, while for the CM, the optimum time was 90 min. From the results, it can be stated that the extraction process by NE and its subsequent purification developed in this study is a simple, economical, environmentally friendly method for producing steviol glycosides. Therefore, this method constitutes a valid alternative to conventional extraction by reducing the extraction time and the consumption of toxic solvents and favouring the use of the extracted metabolites as food additives and/or nutraceuticals. As an added value and of local interest, the experiment was carried out on stevia leaves from the Benevento area (Italy), where a high content of rebaudioside A was observed, which exhibits a sweet taste compared to stevioside, which has a significant bitter aftertaste.

  14. Rapid and efficient method to extract metagenomic DNA from estuarine sediments.

    PubMed

    Shamim, Kashif; Sharma, Jaya; Dubey, Santosh Kumar

    2017-07-01

    Metagenomic DNA from sediments of selective estuaries of Goa, India was extracted using a simple, fast, efficient and environment friendly method. The recovery of pure metagenomic DNA from our method was significantly high as compared to other well-known methods since the concentration of recovered metagenomic DNA ranged from 1185.1 to 4579.7 µg/g of sediment. The purity of metagenomic DNA was also considerably high as the ratio of absorbance at 260 and 280 nm ranged from 1.88 to 1.94. Therefore, the recovered metagenomic DNA was directly used to perform various molecular biology experiments viz. restriction digestion, PCR amplification, cloning and metagenomic library construction. This clearly proved that our protocol for metagenomic DNA extraction using silica gel efficiently removed the contaminants and prevented shearing of the metagenomic DNA. Thus, this modified method can be used to recover pure metagenomic DNA from various estuarine sediments in a rapid, efficient and eco-friendly manner.

  15. Pomegranate extract exhibits in vitro activity against Clostridium difficile.

    PubMed

    Finegold, Sydney M; Summanen, Paula H; Corbett, Karen; Downes, Julia; Henning, Susanne M; Li, Zhaoping

    2014-10-01

    To determine the possible utility of pomegranate extract in the management or prevention of Clostridium difficile infections or colonization. The activity of pomegranate was tested against 29 clinical C. difficile isolates using the Clinical and Laboratory Standards Institute-approved agar dilution technique. Total phenolics content of the pomegranate extract was determined by Folin-Ciocalteau colorimetric method and final concentrations of 6.25 to 400 μg/mL gallic acid equivalent were achieved in the agar. All strains had MICs at 12.5 to 25 mg/mL gallic acid equivalent range. Our results suggest antimicrobial in vitro activity for pomegranate extract against toxigenic C. difficile. Pomegranate extract may be a useful contributor to the management and prevention of C. difficile disease or colonization. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Intra-laboratory validation of chronic bee paralysis virus quantitation using an accredited standardised real-time quantitative RT-PCR method.

    PubMed

    Blanchard, Philippe; Regnault, Julie; Schurr, Frank; Dubois, Eric; Ribière, Magali

    2012-03-01

    Chronic bee paralysis virus (CBPV) is responsible for chronic bee paralysis, an infectious and contagious disease in adult honey bees (Apis mellifera L.). A real-time RT-PCR assay to quantitate the CBPV load is now available. To propose this assay as a reference method, it was characterised further in an intra-laboratory study during which the reliability and the repeatability of results and the performance of the assay were confirmed. The qPCR assay alone and the whole quantitation method (from sample RNA extraction to analysis) were both assessed following the ISO/IEC 17025 standard and the recent XP U47-600 standard issued by the French Standards Institute. The performance of the qPCR assay and of the overall CBPV quantitation method were validated over a 6 log range from 10(2) to 10(8) with a detection limit of 50 and 100 CBPV RNA copies, respectively, and the protocol of the real-time RT-qPCR assay for CBPV quantitation was approved by the French Accreditation Committee. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Preparing silica aerogel monoliths via a rapid supercritical extraction method.

    PubMed

    Carroll, Mary K; Anderson, Ann M; Gorka, Caroline A

    2014-02-28

    A procedure for the fabrication of monolithic silica aerogels in eight hours or less via a rapid supercritical extraction process is described. The procedure requires 15-20 min of preparation time, during which a liquid precursor mixture is prepared and poured into wells of a metal mold that is placed between the platens of a hydraulic hot press, followed by several hours of processing within the hot press. The precursor solution consists of a 1.0:12.0:3.6:3.5 x 10(-3) molar ratio of tetramethylorthosilicate (TMOS):methanol:water:ammonia. In each well of the mold, a porous silica sol-gel matrix forms. As the temperature of the mold and its contents is increased, the pressure within the mold rises. After the temperature/pressure conditions surpass the supercritical point for the solvent within the pores of the matrix (in this case, a methanol/water mixture), the supercritical fluid is released, and monolithic aerogel remains within the wells of the mold. With the mold used in this procedure, cylindrical monoliths of 2.2 cm diameter and 1.9 cm height are produced. Aerogels formed by this rapid method have comparable properties (low bulk and skeletal density, high surface area, mesoporous morphology) to those prepared by other methods that involve either additional reaction steps or solvent extractions (lengthier processes that generate more chemical waste).The rapid supercritical extraction method can also be applied to the fabrication of aerogels based on other precursor recipes.

  18. Preparing Silica Aerogel Monoliths via a Rapid Supercritical Extraction Method

    PubMed Central

    Gorka, Caroline A.

    2014-01-01

    A procedure for the fabrication of monolithic silica aerogels in eight hours or less via a rapid supercritical extraction process is described. The procedure requires 15-20 min of preparation time, during which a liquid precursor mixture is prepared and poured into wells of a metal mold that is placed between the platens of a hydraulic hot press, followed by several hours of processing within the hot press. The precursor solution consists of a 1.0:12.0:3.6:3.5 x 10-3 molar ratio of tetramethylorthosilicate (TMOS):methanol:water:ammonia. In each well of the mold, a porous silica sol-gel matrix forms. As the temperature of the mold and its contents is increased, the pressure within the mold rises. After the temperature/pressure conditions surpass the supercritical point for the solvent within the pores of the matrix (in this case, a methanol/water mixture), the supercritical fluid is released, and monolithic aerogel remains within the wells of the mold. With the mold used in this procedure, cylindrical monoliths of 2.2 cm diameter and 1.9 cm height are produced. Aerogels formed by this rapid method have comparable properties (low bulk and skeletal density, high surface area, mesoporous morphology) to those prepared by other methods that involve either additional reaction steps or solvent extractions (lengthier processes that generate more chemical waste).The rapid supercritical extraction method can also be applied to the fabrication of aerogels based on other precursor recipes. PMID:24637334

  19. Deep liquid-chromatographic purification of uranium extract from technetium

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

    Volk, V.; Dvoeglazov, K; Podrezova, L.

    The recycling of uranium in the nuclear fuel cycle requires the removal of a number of radioactive and stable impurities like {sup 99}Tc from spent fuels. In order to improve the grade of uranium extract purification from technetium the method of liquid chromatography and the apparatus for its performance have been developed. Process of technetium extraction and concentrating in aqueous solution containing reducing agent has been studied on simulated solutions (U-Tc-HNO{sub 3}-30% TBP-isoparM). The dynamic tests of the method have been carried out on the laboratory unit. Solution of diformyl-hydrazine in nitric acid was used as a stationary phase. Silicamore » gel with specific surface of 186 m{sup 2}/g was used as a carrier of the stationary phase. It is shown that the volume of purified extract increases as the solution temperature increases, concentration of reducing agent increases and extract flow rate decreases. It is established that the technetium content in uranium by this method could achieve a value below 0.3 ppm. Some variants of overload and composition of the stationary phase containing the extracted technetium have been offered and tested. It is defined that the method provides reduction of processing medium-active wastes by more than 10 times during finish refining process. (authors)« less

  20. Analytical study to define a helicopter stability derivative extraction method, volume 1

    NASA Technical Reports Server (NTRS)

    Molusis, J. A.

    1973-01-01

    A method is developed for extracting six degree-of-freedom stability and control derivatives from helicopter flight data. Different combinations of filtering and derivative estimate are investigated and used with a Bayesian approach for derivative identification. The combination of filtering and estimate found to yield the most accurate time response match to flight test data is determined and applied to CH-53A and CH-54B flight data. The method found to be most accurate consists of (1) filtering flight test data with a digital filter, followed by an extended Kalman filter (2) identifying a derivative estimate with a least square estimator, and (3) obtaining derivatives with the Bayesian derivative extraction method.

  1. Validation of laboratory-scale recycling test method of paper PSA label products

    Treesearch

    Carl Houtman; Karen Scallon; Richard Oldack

    2008-01-01

    Starting with test methods and a specification developed by the U.S. Postal Service (USPS) Environmentally Benign Pressure Sensitive Adhesive Postage Stamp Program, a laboratory-scale test method and a specification were developed and validated for pressure-sensitive adhesive labels, By comparing results from this new test method and pilot-scale tests, which have been...

  2. [Effects of post-harvest processing and extraction methods on polysaccharides content of Dendrobium officinale].

    PubMed

    Li, Cong; Ning, Li-Dan; Si, Jin-Ping; Wu, Ling-Shang; Liu, Jing-Jing; Song, Xian-Shui; Yu, Qiao-Xian

    2013-02-01

    To reveal the quality variation of polysaccharide in Dendrobium officinale by post-harvest processing and extraction methods, and provide a basis for post-harvest processing and clinical and hygienical applications of Tiepifengdou (Dendrobii Officinalis Caulis). The content of polysaccharides were studied by 4 post-harvest processing methods, i. e. drying by drying closet, drying after scalding by boiling water, drying while twisting, and drying while twisting after scalding by boiling water. And a series of temperatures were set in each processing procedure. An orthogonal test L9 (3(4)) with crushed degrees, solid-liquid ratio, extraction time and extraction times as factors were designed to analyze the dissolution rate of polysaccharides in Tiepifengdou processed by drying while twisting at 80 degrees C. The content of polysaccharides was ranged from 26.59% to 32.70% in different samples processed by different processing methods, among which drying while twisting at 80 degrees C and 100 degrees C respectively were the best. Crushed degree was the most important influence on the dissolution rate of polysaccharides. The dissolution rate of polysaccharides was extremely low when the sample was boiled directly without crushing and sieving. Drying while twisting at 80 degrees C was the best post-harvest processing method, which can help to dry the fresh herbs and improve the accumulation of polysaccharides. Boiling the uncrushed Tiepifengdou for a long time as traditional method could not fully extract polysaccharides, while boiling the crushed Tiepifengdou can efficiently extract polysaccharides.

  3. [Study of blending method for the extracts of herbal plants].

    PubMed

    Liu, Yongsuo; Cao, Min; Chen, Yuying; Hu, Yuzhu; Wang, Yiming; Luo, Guoan

    2006-03-01

    The irregularity in herbal plant composition is influenced by multiple factors. As for quality control of traditional Chinese medicine, the most critical challenge is to ensure the dosage content uniformity. This content uniformity can be improved by blending different batches of the extracts of herbal plants. Nonlinear least-squares regression was used to calculate the blending coefficient, which means no great absolute differences allowed for all ingredients. For traditional Chinese medicines, even relatively smaller differences could present to be very important for all the ingredients. The auto-scaling pretreatment was used prior to the calculation of the blending coefficients. The pretreatment buffered the characteristics of individual data for the ingredients in different batches, so an improved auto-scaling pretreatment method was proposed. With the improved auto-scaling pretreatment, the relative. differences decreased after blending different batches of extracts of herbal plants according to the reference samples. And the content uniformity control of the specific ingredients could be achieved by the error control coefficient. In the studies for the extracts of fructus gardeniae, the relative differences of all the ingredients is less than 3% after blending different batches of the extracts. The results showed that nonlinear least-squares regression can be used to calculate the blending coefficient of the herbal plant extracts.

  4. Student Reciprocal Peer Teaching as a Method for Active Learning: An Experience in an Electrotechnical Laboratory

    NASA Astrophysics Data System (ADS)

    Muñoz-García, Miguel A.; Moreda, Guillermo P.; Hernández-Sánchez, Natalia; Valiño, Vanesa

    2013-10-01

    Active learning is one of the most efficient mechanisms for learning, according to the psychology of learning. When students act as teachers for other students, the communication is more fluent and knowledge is transferred easier than in a traditional classroom. This teaching method is referred to in the literature as reciprocal peer teaching. In this study, the method is applied to laboratory sessions of a higher education institution course, and the students who act as teachers are referred to as "laboratory monitors." A particular way to select the monitors and its impact in the final marks is proposed. A total of 181 students participated in the experiment, experiences with laboratory monitors are discussed, and methods for motivating and training laboratory monitors and regular students are proposed. The types of laboratory sessions that can be led by classmates are discussed. This work is related to the changes in teaching methods in the Spanish higher education system, prompted by the Bologna Process for the construction of the European Higher Education Area

  5. Novel Methods to Generate Active Ingredients-Enriched Ashwagandha Leaves and Extracts.

    PubMed

    Kaul, Sunil C; Ishida, Yoshiyuki; Tamura, Kazuya; Wada, Teruo; Iitsuka, Tomoko; Garg, Sukant; Kim, Mijung; Gao, Ran; Nakai, Shoichi; Okamoto, Youji; Terao, Keiji; Wadhwa, Renu

    2016-01-01

    Ashwagandha (Withania somnifera) is an Ayurvedic herb commonly used in world-renowned traditional Indian home medicine system. Roots of Ashwagandha have been traditionally known to possess a variety of therapeutic and health promoting potentials that have not been sufficiently supported by laboratory studies. Nevertheless, most, if not all, of the preventive and therapeutic potentials have been assigned to its bioactive components, steroidal alkaloids and lactones. In contrast to the traditional use of roots, we have been exploring bioactivities in leaves of Ashwagandha. Here, we report that the leaves possess higher content of active Withanolides, Withaferin-A (Wi-A) and Withanone (Wi-N), as compared to the roots. We also established, for the first time, hydroponic cultivation of Ashwagandha and investigated the effect of various cultivation conditions on the content of Wi-A and Wi-N by chemical analysis and bioassays. We report that the Withanone/Withaferin A-rich leaves could be obtained by manipulating light condition during hydroponic cultivation. Furthermore, we recruited cyclodextrins to prepare extracts with desired ratio of Wi-N and Wi-A. Hydroponically grown Ashwagandha and its extracts with high ratio of withanolides are valuable for cancer treatment.

  6. Novel Methods to Generate Active Ingredients-Enriched Ashwagandha Leaves and Extracts

    PubMed Central

    Kaul, Sunil C.; Ishida, Yoshiyuki; Tamura, Kazuya; Wada, Teruo; Iitsuka, Tomoko; Garg, Sukant; Kim, Mijung; Gao, Ran; Nakai, Shoichi; Okamoto, Youji; Terao, Keiji; Wadhwa, Renu

    2016-01-01

    Ashwagandha (Withania somnifera) is an Ayurvedic herb commonly used in world-renowned traditional Indian home medicine system. Roots of Ashwagandha have been traditionally known to possess a variety of therapeutic and health promoting potentials that have not been sufficiently supported by laboratory studies. Nevertheless, most, if not all, of the preventive and therapeutic potentials have been assigned to its bioactive components, steroidal alkaloids and lactones. In contrast to the traditional use of roots, we have been exploring bioactivities in leaves of Ashwagandha. Here, we report that the leaves possess higher content of active Withanolides, Withaferin-A (Wi-A) and Withanone (Wi-N), as compared to the roots. We also established, for the first time, hydroponic cultivation of Ashwagandha and investigated the effect of various cultivation conditions on the content of Wi-A and Wi-N by chemical analysis and bioassays. We report that the Withanone/Withaferin A-rich leaves could be obtained by manipulating light condition during hydroponic cultivation. Furthermore, we recruited cyclodextrins to prepare extracts with desired ratio of Wi-N and Wi-A. Hydroponically grown Ashwagandha and its extracts with high ratio of withanolides are valuable for cancer treatment. PMID:27936030

  7. Review on the Extraction Methods of Crude oil from all Generation Biofuels in last few Decades

    NASA Astrophysics Data System (ADS)

    Bhargavi, G.; Nageswara Rao, P.; Renganathan, S.

    2018-03-01

    The ever growing demand for the energy fuels, economy of oil, depletion of energy resources and environmental protection are the inevitable challenges required to be solved meticulously in future decades in order to sustain the life of humans and other creatures. Switching to alternate fuels that are renewable, biodegradable, economically and environmentally friendly can quench the minimum thirst of fuel demands, in addition to mitigation of climate changes. At this moment, production of biofuels has got prominence. The term biofuels broadly refer to the fuels derived from living matter either animals or plants. Among the competent biofuels, biodiesel is one of the promising alternates for diesel engines. Biodiesel is renewable, environmentally friendly, safe to use with wide applications and biodegradable. Due to which, it has become a major focus of intensive global research and development of alternate energy. The present review has been focused specifically on biodiesel. Concerning to the biodiesel production, the major steps includes lipid extraction followed by esterification/transesterification. For the extraction of lipids, several extraction techniques have been put forward irrespective of the generations and feed stocks used. This review provides theoretical background on the two major extraction methods, mechanical and chemical extraction methods. The practical issues of each extraction method such as efficiency of extraction, extraction time, oil sources and its pros and cons are discussed. It is conceived that congregating information on oil extraction methods may helpful in further research advancements to ease biofuel production.

  8. Plant Growth Biostimulants Based on Different Methods of Seaweed Extraction with Water.

    PubMed

    Godlewska, Katarzyna; Michalak, Izabela; Tuhy, Łukasz; Chojnacka, Katarzyna

    2016-01-01

    We explored two methods for obtaining aqueous extracts: boiling and soaking of Baltic seaweeds (EB and ES, resp.). Algal extracts were characterized in terms of polyphenols, micro- and macroelements, lipids content, and antibacterial properties. The utilitarian properties were examined in the germination tests on Lepidium sativum for three extract dilutions (0.5, 2.5, and 10%). It was found that the extracts were similar in micro- and macroelement concentrations. Water was proved to be a good solvent to extract phenolic compounds. The algal extract produced by soaking biomass did not show inhibitory effect on Escherichia coli and Staphylococcus aureus. Only the boiled extract had an inhibitory activity against E. coli. Germination tests revealed a positive influence of the bioproducts on the cultivated plants. In the group treated with 10% EB, plants were 13% longer than in the control group; the content of elements B, Mo, Zn, and Na in the group treated with 10% ES was higher by 76%, 48%, 31%, and 59% than in the control group, respectively; the content of chlorophyll was 2.5 times higher in 0.5% ES than in the control group. Extracts showed the slight impact on the morphology of plants.

  9. Plant Growth Biostimulants Based on Different Methods of Seaweed Extraction with Water

    PubMed Central

    Godlewska, Katarzyna; Tuhy, Łukasz; Chojnacka, Katarzyna

    2016-01-01

    We explored two methods for obtaining aqueous extracts: boiling and soaking of Baltic seaweeds (EB and ES, resp.). Algal extracts were characterized in terms of polyphenols, micro- and macroelements, lipids content, and antibacterial properties. The utilitarian properties were examined in the germination tests on Lepidium sativum for three extract dilutions (0.5, 2.5, and 10%). It was found that the extracts were similar in micro- and macroelement concentrations. Water was proved to be a good solvent to extract phenolic compounds. The algal extract produced by soaking biomass did not show inhibitory effect on Escherichia coli and Staphylococcus aureus. Only the boiled extract had an inhibitory activity against E. coli. Germination tests revealed a positive influence of the bioproducts on the cultivated plants. In the group treated with 10% EB, plants were 13% longer than in the control group; the content of elements B, Mo, Zn, and Na in the group treated with 10% ES was higher by 76%, 48%, 31%, and 59% than in the control group, respectively; the content of chlorophyll was 2.5 times higher in 0.5% ES than in the control group. Extracts showed the slight impact on the morphology of plants. PMID:27366749

  10. PRN 96-1: Tolerance Enforcement Methods - Independent Laboratory Validation by Petitioner

    EPA Pesticide Factsheets

    This notice is intended to clarify the requirements for submission of an Independent Laboratory Validation to accompany new pesticide analytical methods and does not contain additional data requirements.This notice supersedes PR Notice 88-5.

  11. Development and validation of an automated liquid-liquid extraction GC/MS method for the determination of THC, 11-OH-THC, and free THC-carboxylic acid (THC-COOH) from blood serum.

    PubMed

    Purschke, Kirsten; Heinl, Sonja; Lerch, Oliver; Erdmann, Freidoon; Veit, Florian

    2016-06-01

    The analysis of Δ(9)-tetrahydrocannabinol (THC) and its metabolites 11-hydroxy-Δ(9)-tetrahydrocannabinol (11-OH-THC), and 11-nor-9-carboxy-Δ(9)-tetrahydrocannabinol (THC-COOH) from blood serum is a routine task in forensic toxicology laboratories. For examination of consumption habits, the concentration of the phase I metabolite THC-COOH is used. Recommendations for interpretation of analysis values in medical-psychological assessments (regranting of driver's licenses, Germany) include threshold values for the free, unconjugated THC-COOH. Using a fully automated two-step liquid-liquid extraction, THC, 11-OH-THC, and free, unconjugated THC-COOH were extracted from blood serum, silylated with N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), and analyzed by GC/MS. The automation was carried out by an x-y-z sample robot equipped with modules for shaking, centrifugation, and solvent evaporation. This method was based on a previously developed manual sample preparation method. Validation guidelines of the Society of Toxicological and Forensic Chemistry (GTFCh) were fulfilled for both methods, at which the focus of this article is the automated one. Limits of detection and quantification for THC were 0.3 and 0.6 μg/L, for 11-OH-THC were 0.1 and 0.8 μg/L, and for THC-COOH were 0.3 and 1.1 μg/L, when extracting only 0.5 mL of blood serum. Therefore, the required limit of quantification for THC of 1 μg/L in driving under the influence of cannabis cases in Germany (and other countries) can be reached and the method can be employed in that context. Real and external control samples were analyzed, and a round robin test was passed successfully. To date, the method is employed in the Institute of Legal Medicine in Giessen, Germany, in daily routine. Automation helps in avoiding errors during sample preparation and reduces the workload of the laboratory personnel. Due to its flexibility, the analysis system can be employed for other liquid-liquid extractions as

  12. LABORATORY EVALUATION OF METHODS TO SEPARATE FINE GRAINED SEDIMENT FROM STORM WATER

    EPA Science Inventory

    A literature survey had been conducted by the St. Anthony Falls Hydraulic laboratory to assess various methods for separation of sediment from storm water at construction sites. Two methods have shown some promise in this application, and a research program was initiated with the...

  13. Revisiting chlorophyll extraction methods in biological soil crusts - methodology for determination of chlorophyll a and chlorophyll a + b as compared to previous methods

    NASA Astrophysics Data System (ADS)

    Caesar, Jennifer; Tamm, Alexandra; Ruckteschler, Nina; Lena Leifke, Anna; Weber, Bettina

    2018-03-01

    Chlorophyll concentrations of biological soil crust (biocrust) samples are commonly determined to quantify the relevance of photosynthetically active organisms within these surface soil communities. Whereas chlorophyll extraction methods for freshwater algae and leaf tissues of vascular plants are well established, there is still some uncertainty regarding the optimal extraction method for biocrusts, where organism composition is highly variable and samples comprise major amounts of soil. In this study we analyzed the efficiency of two different chlorophyll extraction solvents, the effect of grinding the soil samples prior to the extraction procedure, and the impact of shaking as an intermediate step during extraction. The analyses were conducted on four different types of biocrusts. Our results show that for all biocrust types chlorophyll contents obtained with ethanol were significantly lower than those obtained using dimethyl sulfoxide (DMSO) as a solvent. Grinding of biocrust samples prior to analysis caused a highly significant decrease in chlorophyll content for green algal lichen- and cyanolichen-dominated biocrusts, and a tendency towards lower values for moss- and algae-dominated biocrusts. Shaking of the samples after each extraction step had a significant positive effect on the chlorophyll content of green algal lichen- and cyanolichen-dominated biocrusts. Based on our results we confirm a DMSO-based chlorophyll extraction method without grinding pretreatment and suggest the addition of an intermediate shaking step for complete chlorophyll extraction (see Supplement S6 for detailed manual). Determination of a universal chlorophyll extraction method for biocrusts is essential for the inter-comparability of publications conducted across all continents.

  14. Cepstrum based feature extraction method for fungus detection

    NASA Astrophysics Data System (ADS)

    Yorulmaz, Onur; Pearson, Tom C.; Çetin, A. Enis

    2011-06-01

    In this paper, a method for detection of popcorn kernels infected by a fungus is developed using image processing. The method is based on two dimensional (2D) mel and Mellin-cepstrum computation from popcorn kernel images. Cepstral features that were extracted from popcorn images are classified using Support Vector Machines (SVM). Experimental results show that high recognition rates of up to 93.93% can be achieved for both damaged and healthy popcorn kernels using 2D mel-cepstrum. The success rate for healthy popcorn kernels was found to be 97.41% and the recognition rate for damaged kernels was found to be 89.43%.

  15. Extraction and labeling methods for microarrays using small amounts of plant tissue.

    PubMed

    Stimpson, Alexander J; Pereira, Rhea S; Kiss, John Z; Correll, Melanie J

    2009-03-01

    Procedures were developed to maximize the yield of high-quality RNA from small amounts of plant biomass for microarrays. Two disruption techniques (bead milling and pestle and mortar) were compared for the yield and the quality of RNA extracted from 1-week-old Arabidopsis thaliana seedlings (approximately 0.5-30 mg total biomass). The pestle and mortar method of extraction showed enhanced RNA quality at the smaller biomass samples compared with the bead milling technique, although the quality in the bead milling could be improved with additional cooling steps. The RNA extracted from the pestle and mortar technique was further tested to determine if the small quantity of RNA (500 ng-7 microg) was appropriate for microarray analyses. A new method of low-quantity RNA labeling for microarrays (NuGEN Technologies, Inc.) was used on five 7-day-old seedlings (approximately 2.5 mg fresh weight total) of Arabidopsis that were grown in the dark and exposed to 1 h of red light or continued dark. Microarray analyses were performed on a small plant sample (five seedlings; approximately 2.5 mg) using these methods and compared with extractions performed with larger biomass samples (approximately 500 roots). Many well-known light-regulated genes between the small plant samples and the larger biomass samples overlapped in expression changes, and the relative expression levels of selected genes were confirmed with quantitative real-time polymerase chain reaction, suggesting that these methods can be used for plant experiments where the biomass is extremely limited (i.e. spaceflight studies).

  16. Comparison of the DNA extraction methods for polymerase chain reaction amplification from formalin-fixed and paraffin-embedded tissues.

    PubMed

    Sato, Y; Sugie, R; Tsuchiya, B; Kameya, T; Natori, M; Mukai, K

    2001-12-01

    To obtain an adequate quality and quantity of DNA from formalin-fixed and paraffin-embedded tissue, six different DNA extraction methods were compared. Four methods used deparaffinization by xylene followed by proteinase K digestion and phenol-chloroform extraction. The temperature of the different steps was changed to obtain higher yields and improved quality of extracted DNA. The remaining two methods used microwave heating for deparaffinization. The best DNA extraction method consisted of deparaffinization by microwave irradiation, protein digestion with proteinase K at 48 degrees C overnight, and no further purification steps. By this method, the highest DNA yield was obtained and the amplification of a 989-base pair beta-globin gene fragment was achieved. Furthermore, DNA extracted by means of this procedure from five gastric carcinomas was successfully used for single strand conformation polymorphism and direct sequencing assays of the beta-catenin gene. Because the microwave-based DNA extraction method presented here is simple, has a lower contamination risk, and results in a higher yield of DNA compared with the ordinary organic chemical reagent-based extraction method, it is considered applicable to various clinical and basic fields.

  17. Method for the determination of catechin and epicatechin enantiomers in cocoa-based ingredients and products by high-performance liquid chromatography: single-laboratory validation.

    PubMed

    Machonis, Philip R; Jones, Matthew A; Schaneberg, Brian T; Kwik-Uribe, Catherine L

    2012-01-01

    A single-laboratory validation study was performed for an HPLC method to identify and quantify the flavanol enantiomers (+)- and (-)-epicatechin and (+)- and (-)-catechin in cocoa-based ingredients and products. These compounds were eluted isocratically with an ammonium acetate-methanol mobile phase applied to a modified beta-cyclodextrin chiral stationary phase and detected using fluorescence. Spike recovery experiments using appropriate matrix blanks, along with cocoa extract, cocoa powder, and dark chocolate, were used to evaluate accuracy, repeatability, specificity, LOD, LOQ, and linearity of the method as performed by a single analyst on multiple days. In all samples analyzed, (-)-epicatechin was the predominant flavanol and represented 68-91% of the total monomeric flavanols detected. For the cocoa-based products, within-day (intraday) precision for (-)-epicatechin was between 1.46-3.22%, for (+)-catechin between 3.66-6.90%, and for (-)-catechin between 1.69-6.89%; (+)-epicatechin was not detected in these samples. Recoveries for the three sample types investigated ranged from 82.2 to 102.1% at the 50% spiking level, 83.7 to 102.0% at the 100% spiking level, and 80.4 to 101.1% at the 200% spiking level. Based on performance results, this method may be suitable for routine laboratory use in analysis of cocoa-based ingredients and products.

  18. [A customized method for information extraction from unstructured text data in the electronic medical records].

    PubMed

    Bao, X Y; Huang, W J; Zhang, K; Jin, M; Li, Y; Niu, C Z

    2018-04-18

    There is a huge amount of diagnostic or treatment information in electronic medical record (EMR), which is a concrete manifestation of clinicians actual diagnosis and treatment details. Plenty of episodes in EMRs, such as complaints, present illness, past history, differential diagnosis, diagnostic imaging, surgical records, reflecting details of diagnosis and treatment in clinical process, adopt Chinese description of natural language. How to extract effective information from these Chinese narrative text data, and organize it into a form of tabular for analysis of medical research, for the practical utilization of clinical data in the real world, is a difficult problem in Chinese medical data processing. Based on the EMRs narrative text data in a tertiary hospital in China, a customized information extracting rules learning, and rule based information extraction methods is proposed. The overall method consists of three steps, which includes: (1) Step 1, a random sample of 600 copies (including the history of present illness, past history, personal history, family history, etc.) of the electronic medical record data, was extracted as raw corpora. With our developed Chinese clinical narrative text annotation platform, the trained clinician and nurses marked the tokens and phrases in the corpora which would be extracted (with a history of diabetes as an example). (2) Step 2, based on the annotated corpora clinical text data, some extraction templates were summarized and induced firstly. Then these templates were rewritten using regular expressions of Perl programming language, as extraction rules. Using these extraction rules as basic knowledge base, we developed extraction packages in Perl, for extracting data from the EMRs text data. In the end, the extracted data items were organized in tabular data format, for later usage in clinical research or hospital surveillance purposes. (3) As the final step of the method, the evaluation and validation of the proposed

  19. Carbon coated magnetic nanoparticles as a novel magnetic solid phase extraction adsorbent for simultaneous extraction of methamphetamine and ephedrine from urine samples.

    PubMed

    Taghvimi, Arezou; Hamishehkar, Hamed

    2017-01-15

    This paper develops a highly selective, specific and efficient method for simultaneous determination of ephedrine and methamphetamine by a new carbon coated magnetic nanoparticles (C/MNPs) as a magnetic solid phase extraction (MSPE) adsorbent in biological urine medium. The characterization of synthesized magnetic nano adsorbent was completely carried out by various characterization techniques like Fourier transform infrared (FT-IR) spectroscopy, powder x-ray diffraction (XRD), scanning electron microscopy (SEM) and vibrating sample magnetometer (VSM). Nine important parameters influencing extraction efficiency including amount of adsorbent, amounts of sample volume, pH, type and amount of extraction organic solvent, time of extraction and desorption, agitation rate and ionic strength of extraction medium, were studied and optimized. Under optimized extraction conditions, a good linearity was observed in the concentration range of 100-2000ng/mL for ephedrine and 100-2500ng/mL for methamphetamine. Analysis of positive urine samples was carried out by proposed method with the recovery of 98.71 and 97.87% for ephedrine and methamphetamine, respectively. The results indicated that carbon coated magnetic nanoparticles could be applied in clinical and forensic laboratories for simultaneous determination of abused drugs in urine media. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Validated method for determination of eight banned nitroimidazole residues in natural casings by LC/MS/MS with solid-phase extraction.

    PubMed

    Sun, Hanwen; Wang, Fengchi; Ai, Lianfeng; Guo, Chunhai; Chen, Ruichun

    2009-01-01

    A sensitive method based on solid-phase extraction-liquid chromatography-tandem mass spectrometry interfaced with electrospray ionization (SPE-LC-MS/MS-ESI) was developed for the simultaneous determination of 8 banned nitroimidazole (NOZ) drugs including metronidazole (MNZ), ronidazole (RNZ), dimetridazole (DMZ), tinidazole, ornidazole, secnidazole, metronidazole-OH (MNZOH, the metabolite of MNZ), and 2-hydroxymethyl-1-methyl-5-nitroimidazole (HMMNI, the metabolite of RNZ and DMZ) in natural casings. After extraction with ethyl acetate and evaporation, the NOZs were reconstituted in ethyl acetate and purified on a strong cation-exchange SPE column, and then LC/MS/MS analysis was performed by positive ESI applying multiple reaction monitoring of 2 transition reactions for each compound. The method was validated according to the European Union requirements (Commission Decision 2002/657/EC). Specificity, linearity, decision limit (CCalpha), detection capability (CCbeta), accuracy, and precision were determined. Average recoveries of the 8 NOZs from natural animal casing fortified at 3 levels (0.1, 0.5, and 1 microg/kg) ranged from 87.3 to 116.5%. The calculated CCalpha for NOZs ranged from 0.029 to 0.049 microg/kg, and CCbeta ranged from 0.049 to 0.083 microg/kg. Repeatability was in the range of 3.35-10.1%, and within-laboratory reproducibility was <10.3%.

  1. Comparison of Methods for the Extraction of DNA from Formalin-Fixed, Paraffin-Embedded Archival Tissues

    PubMed Central

    Sengüven, Burcu; Baris, Emre; Oygur, Tulin; Berktas, Mehmet

    2014-01-01

    Aim: Discussing a protocol involving xylene-ethanol deparaffinization on slides followed by a kit-based extraction that allows for the extraction of high quality DNA from FFPE tissues. Methods: DNA was extracted from the FFPE tissues of 16 randomly selected blocks. Methods involving deparaffinization on slides or tubes, enzyme digestion overnight or for 72 hours and isolation using phenol chloroform method or a silica-based commercial kit were compared in terms of yields, concentrations and the amplifiability. Results: The highest yield of DNA was produced from the samples that were deparaffinized on slides, digested for 72 hours and isolated with a commercial kit. Samples isolated with the phenol-chloroform method produced DNA of lower purity than the samples that were purified with kit. The samples isolated with the commercial kit resulted in better PCR amplification. Conclusion: Silica-based commercial kits and deparaffinized on slides should be considered for DNA extraction from FFPE. PMID:24688314

  2. Ultrasound assisted methods for enhanced extraction of phycobiliproteins from marine macro-algae, Gelidium pusillum (Rhodophyta).

    PubMed

    Mittal, Rochak; Tavanandi, Hrishikesh A; Mantri, Vaibhav A; Raghavarao, K S M S

    2017-09-01

    Extraction of phycobiliproteins (R-phycoerythrin, R-PE and R-phycocyanin, R-PC) from macro-algae is difficult due to the presence of large polysaccharides (agar, cellulose etc.) present in the cell wall which offer major hindrance for cell disruption. The present study is aimed at developing most suitable methodology for the primary extraction of R-PE and R-PC from marine macro-algae, Gelidium pusillum(Stackhouse) Le Jolis. Such extraction of phycobiliproteins by using ultrasonication and other conventional methods such as maceration, maceration in presence of liquid nitrogen, homogenization, and freezing and thawing (alone and in combinations) is reported for the first time. Standardization of ultrasonication for different parameters such as ultrasonication amplitude (60, 90 and 120µm) and ultrasonication time (1, 2, 4, 6, 8 and 10mins) at different temperatures (30, 35 and 40°C) was carried out. Kinetic parameters were estimated for extraction of phycobiliproteins by ultrasonication based on second order mass transfer kinetics. Based on calorimetric measurements, power, ultrasound intensity and acoustic power density were estimated to be 41.97W, 14.81W/cm 2 and 0.419W/cm 3 , respectively. Synergistic effect of ultrasonication was observed when employed in combination with other conventional primary extraction methods. Homogenization in combination with ultrasonication resulted in an enhancement in efficiency by 9.3% over homogenization alone. Similarly, maceration in combination with ultrasonication resulted in an enhancement in efficiency by 31% over maceration alone. Among all the methods employed, maceration in combination with ultrasonication resulted in the highest extraction efficiency of 77 and 93% for R-PE and R-PC, respectively followed by homogenization in combination with ultrasonication (69.6% for R-PE and 74.1% for R-PC). HPLC analysis was carried out in order to ensure that R-PE was present in the extract and remained intact even after processing

  3. Comparison of extraction methods for the analysis of natural dyes in historical textiles by high-performance liquid chromatography.

    PubMed

    Valianou, Lemonia; Karapanagiotis, Ioannis; Chryssoulakis, Yannis

    2009-12-01

    Different methods for the extraction of Dactylopius coccus Costa, Rubia tinctorum L., Isatis tinctoria L., Reseda luteola L., Curcuma longa L. and Cotinus coggygria Scop. from wool fibres are investigated using high-performance liquid chromatography with diode array detector (HPLC-DAD). The efficiencies of five extraction methods which include the use of HCl (widely used extraction method), citric acid, oxalic acid, TFA and a combination of HCOOH and EDTA are compared on the basis of the (a) number, (b) relative quantities, measured as HPLC peak areas and (c) signal-to-noise ratios (S/N) of the compounds extracted from the wool substrates. Flavonoid glycosides and curcuminoids contained in R. luteola L. and C. longa L., respectively, according to liquid chromatography with mass spectrometry (LC-MS) identifications, are not detected after treating the fibres with HCl. All the other milder methods are successful in extracting these compounds. Experiments are performed using HPLC-DAD to compare the HPLC peak areas and the S/N of the following extracted compounds: indigotin, indirubin, curcumin, demethoxycurcumin, bisdemethoxycurcumin, fisetin, sulfuretin, luteolin, luteolin-7-O-glucoside, apigenin, carminic acid, alizarin, puruprin and rubiadin. It is shown that the TFA method provides overall the best results as it gives elevated extraction yields except for fisetin, luteolin, apigenin and luteolin-7-O-glucoside and highest S/N except for fisetin and luteolin-7-O-glucoside. It is noteworthy that treatment of the fibres with the typical HCl extraction method results overall in very low S/N. The TFA method is selected for further studies, as follows. First, it is applied on silk dyed samples and compared with the HCl method. The same relative differences of the TFA and HCl methods observed for the wool dyed samples are reported for the silk dyed samples too, except for rubiadin, luteolin and apigenin. Thus, in most cases, the nature of the substrate (wool or silk

  4. Comparative efficiency of different methods of gluten extraction in indigenous varieties of wheat.

    PubMed

    Imran, Samra; Hussain, Zaib; Ghafoor, Farkhanda; Nagra, Saeedahmad; Ziai, Naheeda Ashbeal

    2013-06-01

    The present study investigated six varieties of locally grown wheat (Lasani, Sehar, Miraj-08, Chakwal-50, Faisalabad-08 and Inqlab) procured from Punjab Seed Corporation, Lahore, Pakistan for their proximate contents. On the basis of protein content and ready availability, Faisalabad-08 (FD-08) was selected to be used for the assessment of comparative efficiency of various methods used for gluten extraction. Three methods, mechanical, chemical and microbiological were used for the extraction of gluten from FD-08. Each method was carried out under ambient conditions using a drying temperature of 55 degrees C. Mechanical method utilized four different processes viz:- dough process, dough batter process, batter process and ethanol washing process using standard 150 mesh. The starch thus obtained was analyzed for its proximate contents. Dough batter process proved to be the most efficient mechanical method and was further investigated using 200 and 300 mesh. Gluten content was determined using sandwich omega-gliadin enzyme-linked immunosorbent assay (ELISA).The results of dough batter process using 200 mesh indicated a starch product with gluten content of 678 ppm. Chemical method indicated high gluten content of more than 5000 ppm and the microbiological method reduced the gluten content from 2500 ppm to 398 ppm. From the results it was observed that no gluten extraction method is viable to produce starch which can fulfill the criteria of a gluten free product (20 ppm).

  5. A method for the solvent extraction of low-boiling-point plant volatiles.

    PubMed

    Xu, Ning; Gruber, Margaret; Westcott, Neil; Soroka, Julie; Parkin, Isobel; Hegedus, Dwayne

    2005-01-01

    A new method has been developed for the extraction of volatiles from plant materials and tested on seedling tissue and mature leaves of Arabidopsis thaliana, pine needles and commercial mixtures of plant volatiles. Volatiles were extracted with n-pentane and then subjected to quick distillation at a moderate temperature. Under these conditions, compounds such as pigments, waxes and non-volatile compounds remained undistilled, while short-chain volatile compounds were distilled into a receiving flask using a high-efficiency condenser. Removal of the n-pentane and concentration of the volatiles in the receiving flask was carried out using a Vigreux column condenser prior to GC-MS. The method is ideal for the rapid extraction of low-boiling-point volatiles from small amounts of plant material, such as is required when conducting metabolic profiling or defining biological properties of volatile components from large numbers of mutant lines.

  6. 2D automatic body-fitted structured mesh generation using advancing extraction method

    NASA Astrophysics Data System (ADS)

    Zhang, Yaoxin; Jia, Yafei

    2018-01-01

    This paper presents an automatic mesh generation algorithm for body-fitted structured meshes in Computational Fluids Dynamics (CFD) analysis using the Advancing Extraction Method (AEM). The method is applicable to two-dimensional domains with complex geometries, which have the hierarchical tree-like topography with extrusion-like structures (i.e., branches or tributaries) and intrusion-like structures (i.e., peninsula or dikes). With the AEM, the hierarchical levels of sub-domains can be identified, and the block boundary of each sub-domain in convex polygon shape in each level can be extracted in an advancing scheme. In this paper, several examples were used to illustrate the effectiveness and applicability of the proposed algorithm for automatic structured mesh generation, and the implementation of the method.

  7. Development of automated extraction method of biliary tract from abdominal CT volumes based on local intensity structure analysis

    NASA Astrophysics Data System (ADS)

    Koga, Kusuto; Hayashi, Yuichiro; Hirose, Tomoaki; Oda, Masahiro; Kitasaka, Takayuki; Igami, Tsuyoshi; Nagino, Masato; Mori, Kensaku

    2014-03-01

    In this paper, we propose an automated biliary tract extraction method from abdominal CT volumes. The biliary tract is the path by which bile is transported from liver to the duodenum. No extraction method have been reported for the automated extraction of the biliary tract from common contrast CT volumes. Our method consists of three steps including: (1) extraction of extrahepatic bile duct (EHBD) candidate regions, (2) extraction of intrahepatic bile duct (IHBD) candidate regions, and (3) combination of these candidate regions. The IHBD has linear structures and intensities of the IHBD are low in CT volumes. We use a dark linear structure enhancement (DLSE) filter based on a local intensity structure analysis method using the eigenvalues of the Hessian matrix for the IHBD candidate region extraction. The EHBD region is extracted using a thresholding process and a connected component analysis. In the combination process, we connect the IHBD candidate regions to each EHBD candidate region and select a bile duct region from the connected candidate regions. We applied the proposed method to 22 cases of CT volumes. An average Dice coefficient of extraction result was 66.7%.

  8. Optimizing Fungal DNA Extraction Methods from Aerosol Filters

    NASA Astrophysics Data System (ADS)

    Jimenez, G.; Mescioglu, E.; Paytan, A.

    2016-12-01

    Fungi and fungal spores can be picked up from terrestrial ecosystems, transported long distances, and deposited into marine ecosystems. It is important to study dust-borne fungal communities, because they can stay viable and effect the ambient microbial populations, which are key players in biogeochemical cycles. One of the challenges of studying dust-borne fungal populations is that aerosol samples contain low biomass, making extracting good quality DNA very difficult. The aim of this project was to increase DNA yield by optimizing DNA extraction methods. We tested aerosol samples collected from Haifa, Israel (polycarbonate filter), Monterey Bay, CA (quartz filter) and Bermuda (quartz filter). Using the Qiagen DNeasy Plant Kit, we tested the effect of altering bead beating times and incubation times, adding three freeze and thaw steps, initially washing the filters with buffers for various lengths of time before using the kit, and adding a step with 30 minutes of sonication in 65C water. Adding three freeze/thaw steps, adding a sonication step, washing with a phosphate buffered saline overnight, and increasing incubation time to two hours, in that order, resulted in the highest increase in DNA for samples from Israel (polycarbonate). DNA yield of samples from Monterey (quart filter) increased about 5 times when washing with buffers overnight (phosphate buffered saline and potassium phophate buffer), adding a sonication step, and adding three freeze and thaw steps. Samples collected in Bermuda (quartz filter) had the highest increase in DNA yield from increasing incubation to 2 hours, increasing bead beating time to 6 minutes, and washing with buffers overnight (phosphate buffered saline and potassium phophate buffer). Our results show that DNA yield can be increased by altering various steps of the Qiagen DNeasy Plant Kit protocol, but different types of filters collected at different sites respond differently to alterations. These results can be used as

  9. Extraction of CYP chemical interactions from biomedical literature using natural language processing methods.

    PubMed

    Jiao, Dazhi; Wild, David J

    2009-02-01

    This paper proposes a system that automatically extracts CYP protein and chemical interactions from journal article abstracts, using natural language processing (NLP) and text mining methods. In our system, we employ a maximum entropy based learning method, using results from syntactic, semantic, and lexical analysis of texts. We first present our system architecture and then discuss the data set for training our machine learning based models and the methods in building components in our system, such as part of speech (POS) tagging, Named Entity Recognition (NER), dependency parsing, and relation extraction. An evaluation of the system is conducted at the end, yielding very promising results: The POS, dependency parsing, and NER components in our system have achieved a very high level of accuracy as measured by precision, ranging from 85.9% to 98.5%, and the precision and the recall of the interaction extraction component are 76.0% and 82.6%, and for the overall system are 68.4% and 72.2%, respectively.

  10. [Optimization of extraction process for tannins from Geranium orientali-tibeticum by supercritical CO2 method].

    PubMed

    Xie, Song; Tong, Zhi-Ping; Tan, Rui; Liu, Xiao-Zhen

    2014-08-01

    In order to optimize extraction process conditions of tannins from Geranium orientali-tibeticum by supercritical CO2, the content of tannins was determined by phosphomolybdium tungsten acid-casein reaction, with extraction pressure, extraction temper- ature and extraction time as factors, the content of tannins from extract of G. orientali-tibeticum as index, technology conditions were optimized by orthogonal test. Optimum technology conditions were as follows: extraction pressure was 25 MPa, extraction temperature was 50 °C, extracted 1.5 h. The content of tannins in extract was 12.91 mg x g(-1), extract rate was 3.67%. The method established could be used for assay the contents of tannin in G. orientali-tibeticum. The circulated extraction was an effective extraction process that was stable and feasible, and that provides a way of the extraction process conditions of tannin from G. orientali-tibeticum.

  11. A window-based time series feature extraction method.

    PubMed

    Katircioglu-Öztürk, Deniz; Güvenir, H Altay; Ravens, Ursula; Baykal, Nazife

    2017-10-01

    This study proposes a robust similarity score-based time series feature extraction method that is termed as Window-based Time series Feature ExtraCtion (WTC). Specifically, WTC generates domain-interpretable results and involves significantly low computational complexity thereby rendering itself useful for densely sampled and populated time series datasets. In this study, WTC is applied to a proprietary action potential (AP) time series dataset on human cardiomyocytes and three precordial leads from a publicly available electrocardiogram (ECG) dataset. This is followed by comparing WTC in terms of predictive accuracy and computational complexity with shapelet transform and fast shapelet transform (which constitutes an accelerated variant of the shapelet transform). The results indicate that WTC achieves a slightly higher classification performance with significantly lower execution time when compared to its shapelet-based alternatives. With respect to its interpretable features, WTC has a potential to enable medical experts to explore definitive common trends in novel datasets. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Development of Novel Method for Rapid Extract of Radionuclides from Solution Using Polymer Ligand Film

    NASA Astrophysics Data System (ADS)

    Rim, Jung H.

    Accurate and fast determination of the activity of radionuclides in a sample is critical for nuclear forensics and emergency response. Radioanalytical techniques are well established for radionuclides measurement, however, they are slow and labor intensive, requiring extensive radiochemical separations and purification prior to analysis. With these limitations of current methods, there is great interest for a new technique to rapidly process samples. This dissertation describes a new analyte extraction medium called Polymer Ligand Film (PLF) developed to rapidly extract radionuclides. Polymer Ligand Film is a polymer medium with ligands incorporated in its matrix that selectively and rapidly extract analytes from a solution. The main focus of the new technique is to shorten and simplify the procedure necessary to chemically isolate radionuclides for determination by alpha spectrometry or beta counting. Five different ligands were tested for plutonium extraction: bis(2-ethylhexyl) methanediphosphonic acid (H2DEH[MDP]), di(2-ethyl hexyl) phosphoric acid (HDEHP), trialkyl methylammonium chloride (Aliquat-336), 4,4'(5')-di-t-butylcyclohexano 18-crown-6 (DtBuCH18C6), and 2-ethylhexyl 2-ethylhexylphosphonic acid (HEH[EHP]). The ligands that were effective for plutonium extraction further studied for uranium extraction. The plutonium recovery by PLFs has shown dependency on nitric acid concentration and ligand to total mass ratio. H2DEH[MDP] PLFs performed best with 1:10 and 1:20 ratio PLFs. 50.44% and 47.61% of plutonium were extracted on the surface of PLFs with 1M nitric acid for 1:10 and 1:20 PLF, respectively. HDEHP PLF provided the best combination of alpha spectroscopy resolution and plutonium recovery with 1:5 PLF when used with 0.1M nitric acid. The overall analyte recovery was lower than electrodeposited samples, which typically has recovery above 80%. However, PLF is designed to be a rapid field deployable screening technique and consistency is more important

  13. Automated methods of tree boundary extraction and foliage transparency estimation from digital imagery

    Treesearch

    Sang-Mook Lee; Neil A. Clark; Philip A. Araman

    2003-01-01

    Foliage transparency in trees is an important indicator for forest health assessment. This paper helps advance transparency measurement research by presenting methods of automatic tree boundary extraction and foliage transparency estimation from digital images taken from the ground of open grown trees.Extraction of proper boundaries of tree crowns is the...

  14. Do Toxicity Identification and Evaluation Laboratory-Based Methods Reflect Causes of Field Impairment?

    EPA Science Inventory

    Sediment Toxicity Identification and Evaluation (TIE) methods have been developed for both interstitial waters and whole sediments. These relatively simple laboratory methods are designed to identify specific toxicants or classes of toxicants in sediments; however, the question ...

  15. Method development for the analysis of N-nitrosodimethylamine and other N-nitrosamines in drinking water at low nanogram/liter concentrations using solid-phase extraction and gas chromatography with chemical ionization tandem mass spectrometry.

    PubMed

    Munch, Jean W; Bassett, Margarita V

    2006-01-01

    N-nitrosodimethylamine (NDMA) is a probable human carcinogen of concern that has been identified as a drinking water contaminant. U.S. Environmental Protection Agency Method 521 has been developed for the analysis of NDMA and 6 additional N-nitrosamines in drinking water at low ng/L concentrations. The method uses solid-phase extraction with coconut charcoal as the sorbent and dichloromethane as the eluent to concentrate 0.50 L water samples to 1 mL. The extracts are analyzed by gas chromatography-chemical ionization tandem mass spectrometry using large-volume injection. Method performance was evaluated in 2 laboratories. Typical analyte recoveries of 87-104% were demonstrated for fortified reagent water samples, and recoveries of 77-106% were demonstrated for fortified drinking water samples. All relative standard deviations on replicate analyses were < 11%.

  16. Physicochemical and Antioxidant Properties of Rice Bran Oils Produced from Colored Rice Using Different Extraction Methods.

    PubMed

    Mingyai, Sukanya; Kettawan, Aikkarach; Srikaeo, Khongsak; Singanusong, Riantong

    2017-06-01

    This study investigated the physicochemical and antioxidant properties of rice bran oil (RBO) produced from the bran of three rice varities; Khao Dawk Mali 105 (white rice), Red Jasmine rice (red rice) and Hom-nin rice (black rice) using three extraction methods including cold-press extraction (CPE), solvent extraction (SE) and supercritical CO 2 extraction (SC-CO 2 ). Yields, color, acid value (AV), free fatty acid (FFA), peroxide value (PV), iodine value (IV), total phenolic compound (TPC), γ-oryzanol, α-tocopherol and fatty acid profile were analyzed. It was found that the yields obtained from SE, SC-CO 2 and CPE extractions were 17.35-20.19%, 14.76-18.16% and 3.22-6.22%, respectively. The RBO from the bran of red and black rice samples exhibited high antioxidant activities. They also contained higher amount of γ-oryzanol and α-tocopherol than those of white rice sample. In terms of extraction methods, SC-CO 2 provided better qualities of RBO as evidenced by their physicochemical and antioxidant properties. This study found that RBO produced from the bran of black rice samples using SC-CO 2 extraction method showed the best physicochemical and antioxidant properties.

  17. SRC-I demonstration plant analytical laboratory methods manual. Final technical report

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

    Klusaritz, M.L.; Tewari, K.C.; Tiedge, W.F.

    1983-03-01

    This manual is a compilation of analytical procedures required for operation of a Solvent-Refined Coal (SRC-I) demonstration or commercial plant. Each method reproduced in full includes a detailed procedure, a list of equipment and reagents, safety precautions, and, where possible, a precision statement. Procedures for the laboratory's environmental and industrial hygiene modules are not included. Required American Society for Testing and Materials (ASTM) methods are cited, and ICRC's suggested modifications to these methods for handling coal-derived products are provided.

  18. Novel method of extracting motion from natural movies.

    PubMed

    Suzuki, Wataru; Ichinohe, Noritaka; Tani, Toshiki; Hayami, Taku; Miyakawa, Naohisa; Watanabe, Satoshi; Takeichi, Hiroshige

    2017-11-01

    The visual system in primates can be segregated into motion and shape pathways. Interaction occurs at multiple stages along these pathways. Processing of shape-from-motion and biological motion is considered to be a higher-order integration process involving motion and shape information. However, relatively limited types of stimuli have been used in previous studies on these integration processes. We propose a new algorithm to extract object motion information from natural movies and to move random dots in accordance with the information. The object motion information is extracted by estimating the dynamics of local normal vectors of the image intensity projected onto the x-y plane of the movie. An electrophysiological experiment on two adult common marmoset monkeys (Callithrix jacchus) showed that the natural and random dot movies generated with this new algorithm yielded comparable neural responses in the middle temporal visual area. In principle, this algorithm provided random dot motion stimuli containing shape information for arbitrary natural movies. This new method is expected to expand the neurophysiological and psychophysical experimental protocols to elucidate the integration processing of motion and shape information in biological systems. The novel algorithm proposed here was effective in extracting object motion information from natural movies and provided new motion stimuli to investigate higher-order motion information processing. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

  19. Evaluating the Impact of DNA Extraction Method on the Representation of Human Oral Bacterial and Fungal Communities

    PubMed Central

    Biswas, Kristi; Taylor, Michael W.; Gear, Kim

    2017-01-01

    The application of high-throughput, next-generation sequencing technologies has greatly improved our understanding of the human oral microbiome. While deciphering this diverse microbial community using such approaches is more accurate than traditional culture-based methods, experimental bias introduced during critical steps such as DNA extraction may compromise the results obtained. Here, we systematically evaluate four commonly used microbial DNA extraction methods (MoBio PowerSoil® DNA Isolation Kit, QIAamp® DNA Mini Kit, Zymo Bacterial/Fungal DNA Mini PrepTM, phenol:chloroform-based DNA isolation) based on the following criteria: DNA quality and yield, and microbial community structure based on Illumina amplicon sequencing of the V3–V4 region of the 16S rRNA gene of bacteria and the internal transcribed spacer (ITS) 1 region of fungi. Our results indicate that DNA quality and yield varied significantly with DNA extraction method. Representation of bacterial genera in plaque and saliva samples did not significantly differ across DNA extraction methods and DNA extraction method showed no effect on the recovery of fungal genera from plaque. By contrast, fungal diversity from saliva was affected by DNA extraction method, suggesting that not all protocols are suitable to study the salivary mycobiome. PMID:28099455

  20. Method of Grassland Information Extraction Based on Multi-Level Segmentation and Cart Model

    NASA Astrophysics Data System (ADS)

    Qiao, Y.; Chen, T.; He, J.; Wen, Q.; Liu, F.; Wang, Z.

    2018-04-01

    It is difficult to extract grassland accurately by traditional classification methods, such as supervised method based on pixels or objects. This paper proposed a new method combing the multi-level segmentation with CART (classification and regression tree) model. The multi-level segmentation which combined the multi-resolution segmentation and the spectral difference segmentation could avoid the over and insufficient segmentation seen in the single segmentation mode. The CART model was established based on the spectral characteristics and texture feature which were excavated from training sample data. Xilinhaote City in Inner Mongolia Autonomous Region was chosen as the typical study area and the proposed method was verified by using visual interpretation results as approximate truth value. Meanwhile, the comparison with the nearest neighbor supervised classification method was obtained. The experimental results showed that the total precision of classification and the Kappa coefficient of the proposed method was 95 % and 0.9, respectively. However, the total precision of classification and the Kappa coefficient of the nearest neighbor supervised classification method was 80 % and 0.56, respectively. The result suggested that the accuracy of classification proposed in this paper was higher than the nearest neighbor supervised classification method. The experiment certificated that the proposed method was an effective extraction method of grassland information, which could enhance the boundary of grassland classification and avoid the restriction of grassland distribution scale. This method was also applicable to the extraction of grassland information in other regions with complicated spatial features, which could avoid the interference of woodland, arable land and water body effectively.

  1. A simple and rapid infrared-assisted self enzymolysis extraction method for total flavonoid aglycones extraction from Scutellariae Radix and mechanism exploration.

    PubMed

    Wang, Liping; Duan, Haotian; Jiang, Jiebing; Long, Jiakun; Yu, Yingjia; Chen, Guiliang; Duan, Gengli

    2017-09-01

    A new, simple, and fast infrared-assisted self enzymolysis extraction (IRASEE) approach for the extraction of total flavonoid aglycones (TFA) mainly including baicalein, wogonin, and oroxylin A from Scutellariae Radix is presented to enhance extraction yield. Extraction enzymolysis temperature, enzymolysis liquid-to-solid ratio, enzymolysis pH, enzymolysis time and infrared power, the factors affecting IRASEE procedure, were investigated in a newly designed, temperature-controlled infrared-assisted extraction (TC-IRAE) system to acquire the optimum analysis conditions. The results illustrated that IRASEE possessed great advantages in terms of efficiency and time compared with other conventional extraction techniques. Furthermore, the mechanism of IRASEE was preliminarily explored by observing the microscopic change of the samples surface structures, studying the main chemical compositions change of the samples before and after extraction and investigating the kinetics and thermodynamics at three temperature levels during the IRASEE process. These findings revealed that IRASEE can destroy the surface microstructures to accelerate the mass transfer and reduce the activation energy to intensify the chemical process. This integrative study presents a simple, rapid, efficient, and environmental IRASEE method for TFA extraction which has promising prospects for other similar herbal medicines. Graphical Abstract ᅟ.

  2. Ion binding compounds, radionuclide complexes, methods of making radionuclide complexes, methods of extracting radionuclides, and methods of delivering radionuclides to target locations

    DOEpatents

    Chen, Xiaoyuan; Wai, Chien M.; Fisher, Darrell R.

    2000-01-01

    The invention pertains to compounds for binding lanthanide ions and actinide ions. The invention further pertains to compounds for binding radionuclides, and to methods of making radionuclide complexes. Also, the invention pertains to methods of extracting radionuclides. Additionally, the invention pertains to methods of delivering radionuclides to target locations. In one aspect, the invention includes a compound comprising: a) a calix[n]arene group, wherein n is an integer greater than 3, the calix[n]arene group comprising an upper rim and a lower rim; b) at least one ionizable group attached to the lower rim; and c) an ion selected from the group consisting of lanthanide and actinide elements bound to the ionizable group. In another aspect, the invention includes a method of extracting a radionuclide, comprising: a) providing a sample comprising a radionuclide; b) providing a calix[n]arene compound in contact with the sample, wherein n is an integer greater than 3; and c) extracting radionuclide from the sample into the calix[n]arene compound. In yet another aspect, the invention includes a method of delivering a radionuclide to a target location, comprising: a) providing a calix[n]arene compound, wherein n is an integer greater than 3, the calix[n]arene compound comprising at least one ionizable group; b) providing a radionuclide bound to the calix[n]arene compound; and c) providing an antibody attached to the calix[n]arene compound, the antibody being specific for a material found at the target location.

  3. Texture Analysis and Cartographic Feature Extraction.

    DTIC Science & Technology

    1985-01-01

    Investigations into using various image descriptors as well as developing interactive feature extraction software on the Digital Image Analysis Laboratory...system. Originator-supplied keywords: Ad-Hoc image descriptor; Bayes classifier; Bhattachryya distance; Clustering; Digital Image Analysis Laboratory

  4. An Efficient Method for Automatic Road Extraction Based on Multiple Features from LiDAR Data

    NASA Astrophysics Data System (ADS)

    Li, Y.; Hu, X.; Guan, H.; Liu, P.

    2016-06-01

    The road extraction in urban areas is difficult task due to the complicated patterns and many contextual objects. LiDAR data directly provides three dimensional (3D) points with less occlusions and smaller shadows. The elevation information and surface roughness are distinguishing features to separate roads. However, LiDAR data has some disadvantages are not beneficial to object extraction, such as the irregular distribution of point clouds and lack of clear edges of roads. For these problems, this paper proposes an automatic road centerlines extraction method which has three major steps: (1) road center point detection based on multiple feature spatial clustering for separating road points from ground points, (2) local principal component analysis with least squares fitting for extracting the primitives of road centerlines, and (3) hierarchical grouping for connecting primitives into complete roads network. Compared with MTH (consist of Mean shift algorithm, Tensor voting, and Hough transform) proposed in our previous article, this method greatly reduced the computational cost. To evaluate the proposed method, the Vaihingen data set, a benchmark testing data provided by ISPRS for "Urban Classification and 3D Building Reconstruction" project, was selected. The experimental results show that our method achieve the same performance by less time in road extraction using LiDAR data.

  5. Effect of DNA extraction methods and sampling techniques on the apparent structure of cow and sheep rumen microbial communities.

    PubMed

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J; Waghorn, Garry C; Janssen, Peter H

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  6. Effect of DNA Extraction Methods and Sampling Techniques on the Apparent Structure of Cow and Sheep Rumen Microbial Communities

    PubMed Central

    Henderson, Gemma; Cox, Faith; Kittelmann, Sandra; Miri, Vahideh Heidarian; Zethof, Michael; Noel, Samantha J.; Waghorn, Garry C.; Janssen, Peter H.

    2013-01-01

    Molecular microbial ecology techniques are widely used to study the composition of the rumen microbiota and to increase understanding of the roles they play. Therefore, sampling and DNA extraction methods that result in adequate yields of microbial DNA that also accurately represents the microbial community are crucial. Fifteen different methods were used to extract DNA from cow and sheep rumen samples. The DNA yield and quality, and its suitability for downstream PCR amplifications varied considerably, depending on the DNA extraction method used. DNA extracts from nine extraction methods that passed these first quality criteria were evaluated further by quantitative PCR enumeration of microbial marker loci. Absolute microbial numbers, determined on the same rumen samples, differed by more than 100-fold, depending on the DNA extraction method used. The apparent compositions of the archaeal, bacterial, ciliate protozoal, and fungal communities in identical rumen samples were assessed using 454 Titanium pyrosequencing. Significant differences in microbial community composition were observed between extraction methods, for example in the relative abundances of members of the phyla Bacteroidetes and Firmicutes. Microbial communities in parallel samples collected from cows by oral stomach-tubing or through a rumen fistula, and in liquid and solid rumen digesta fractions, were compared using one of the DNA extraction methods. Community representations were generally similar, regardless of the rumen sampling technique used, but significant differences in the abundances of some microbial taxa such as the Clostridiales and the Methanobrevibacter ruminantium clade were observed. The apparent microbial community composition differed between rumen sample fractions, and Prevotellaceae were most abundant in the liquid fraction. DNA extraction methods that involved phenol-chloroform extraction and mechanical lysis steps tended to be more comparable. However, comparison of data

  7. 2D Automatic body-fitted structured mesh generation using advancing extraction method

    USDA-ARS?s Scientific Manuscript database

    This paper presents an automatic mesh generation algorithm for body-fitted structured meshes in Computational Fluids Dynamics (CFD) analysis using the Advancing Extraction Method (AEM). The method is applicable to two-dimensional domains with complex geometries, which have the hierarchical tree-like...

  8. 2D automatic body-fitted structured mesh generation using advancing extraction method

    USDA-ARS?s Scientific Manuscript database

    This paper presents an automatic mesh generation algorithm for body-fitted structured meshes in Computational Fluids Dynamics (CFD) analysis using the Advancing Extraction Method (AEM). The method is applicable to two-dimensional domains with complex geometries, which have the hierarchical tree-like...

  9. Comparison of methods for the extraction of DNA from formalin-fixed, paraffin-embedded archival tissues.

    PubMed

    Sengüven, Burcu; Baris, Emre; Oygur, Tulin; Berktas, Mehmet

    2014-01-01

    Discussing a protocol involving xylene-ethanol deparaffinization on slides followed by a kit-based extraction that allows for the extraction of high quality DNA from FFPE tissues. DNA was extracted from the FFPE tissues of 16 randomly selected blocks. Methods involving deparaffinization on slides or tubes, enzyme digestion overnight or for 72 hours and isolation using phenol chloroform method or a silica-based commercial kit were compared in terms of yields, concentrations and the amplifiability. The highest yield of DNA was produced from the samples that were deparaffinized on slides, digested for 72 hours and isolated with a commercial kit. Samples isolated with the phenol-chloroform method produced DNA of lower purity than the samples that were purified with kit. The samples isolated with the commercial kit resulted in better PCR amplification. Silica-based commercial kits and deparaffinized on slides should be considered for DNA extraction from FFPE.

  10. A simple method for the extraction and identification of light density microplastics from soil.

    PubMed

    Zhang, Shaoliang; Yang, Xiaomei; Gertsen, Hennie; Peters, Piet; Salánki, Tamás; Geissen, Violette

    2018-03-01

    This article introduces a simple and cost-saving method developed to extract, distinguish and quantify light density microplastics of polyethylene (PE) and polypropylene (PP) in soil. A floatation method using distilled water was used to extract the light density microplastics from soil samples. Microplastics and impurities were identified using a heating method (3-5s at 130°C). The number and size of particles were determined using a camera (Leica DFC 425) connected to a microscope (Leica wild M3C, Type S, simple light, 6.4×). Quantification of the microplastics was conducted using a developed model. Results showed that the floatation method was effective in extracting microplastics from soils, with recovery rates of approximately 90%. After being exposed to heat, the microplastics in the soil samples melted and were transformed into circular transparent particles while other impurities, such as organic matter and silicates were not changed by the heat. Regression analysis of microplastics weight and particle volume (a calculation based on image J software analysis) after heating showed the best fit (y=1.14x+0.46, R 2 =99%, p<0.001). Recovery rates based on the empirical model method were >80%. Results from field samples collected from North-western China prove that our method of repetitive floatation and heating can be used to extract, distinguish and quantify light density polyethylene microplastics in soils. Microplastics mass can be evaluated using the empirical model. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. [Comparative study of different extraction methods and assays of tannins in some pteridophytes].

    PubMed

    Laurent, S

    1975-10-01

    Various processes of extraction and quantitative analysis of a condensed tannin in a plant extract, which also includes some chlorogenic acids, have been examined. 60% methanol, at 50 degrees C, proved the most efficient extraction solvent. Several methods of analysis have been tried. The measure of the colour intensity obtained by the action of sulphuric vanilline on flavanols cannot be used because it depends on the tannin condensation stage. It is impossible to separate tannin from chlorogenic acids using the methods of adsorption by skin or nylon powders, or precipitation by polyvinylpyrrolidone. Only paper chromatography, followed by the distinct elution of the various phenolic compounds, allows the tannin evaluation by subtraction; but owing to the variability of the results, many more experiments are necessary. Some other processes are being studied.

  12. A new method for stable lead isotope extraction from seawater.

    PubMed

    Zurbrick, Cheryl M; Gallon, Céline; Flegal, A Russell

    2013-10-24

    A new technique for stable lead (Pb) isotope extraction from seawater is established using Toyopearl AF-Chelate 650M(®) resin (Tosoh Bioscience LLC). This new method is advantageous because it is semi-automated and relatively fast; in addition it introduces a relatively low blank by minimizing the volume of chemicals used in the extraction. Subsequent analyses by HR ICP-MS have a good relative external precision (2σ) of 3.5‰ for (206)Pb/(207)Pb, while analyses by MC-ICP-MS have a better relative external precision of 0.6‰. However, Pb sample concentrations limit MC-ICP-MS analyses to (206)Pb, (207)Pb, and (208)Pb. The method was validated by processing the common Pb isotope reference material NIST SRM-981 and several GEOTRACES intercalibration samples, followed by analyses by HR ICP-MS, all of which showed good agreement with previously reported values. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. An Optimal Control Method for Maximizing the Efficiency of Direct Drive Ocean Wave Energy Extraction System

    PubMed Central

    Chen, Zhongxian; Yu, Haitao; Wen, Cheng

    2014-01-01

    The goal of direct drive ocean wave energy extraction system is to convert ocean wave energy into electricity. The problem explored in this paper is the design and optimal control for the direct drive ocean wave energy extraction system. An optimal control method based on internal model proportion integration differentiation (IM-PID) is proposed in this paper though most of ocean wave energy extraction systems are optimized by the structure, weight, and material. With this control method, the heavy speed of outer heavy buoy of the energy extraction system is in resonance with incident wave, and the system efficiency is largely improved. Validity of the proposed optimal control method is verified in both regular and irregular ocean waves, and it is shown that IM-PID control method is optimal in that it maximizes the energy conversion efficiency. In addition, the anti-interference ability of IM-PID control method has been assessed, and the results show that the IM-PID control method has good robustness, high precision, and strong anti-interference ability. PMID:25152913

  14. An optimal control method for maximizing the efficiency of direct drive ocean wave energy extraction system.

    PubMed

    Chen, Zhongxian; Yu, Haitao; Wen, Cheng

    2014-01-01

    The goal of direct drive ocean wave energy extraction system is to convert ocean wave energy into electricity. The problem explored in this paper is the design and optimal control for the direct drive ocean wave energy extraction system. An optimal control method based on internal model proportion integration differentiation (IM-PID) is proposed in this paper though most of ocean wave energy extraction systems are optimized by the structure, weight, and material. With this control method, the heavy speed of outer heavy buoy of the energy extraction system is in resonance with incident wave, and the system efficiency is largely improved. Validity of the proposed optimal control method is verified in both regular and irregular ocean waves, and it is shown that IM-PID control method is optimal in that it maximizes the energy conversion efficiency. In addition, the anti-interference ability of IM-PID control method has been assessed, and the results show that the IM-PID control method has good robustness, high precision, and strong anti-interference ability.

  15. Method Development for Comprehensive Extraction and Analysis of Marine Toxins: Liquid-Liquid Extraction and Tandem Liquid Chromatography Separations Coupled to Electrospray Tandem Mass Spectrometry

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

    Wunschel, David S.; Valenzuela, Blandina R.; Kaiser, Brooke L. Deatherage

    A variety of toxins are produced by marine and freshwater microorganisms that present a threat to human health. These toxins have diverse chemical properties and specifically, a range of hydrophobicity. Methods for extraction and identification of these toxins are often geared toward specific classes of toxin depending on the sample type. There is a need for a general method of toxin extraction and identification for screening samples where the likely toxin content is not known a priori. Here, we have applied a general method for metabolite extraction to toxin containing samples. This method was coupled with a simple dual liquidmore » chromatography approach for separating a broad range of toxins. This liquid chromatography approach was coupled to triple quadrupole and quadrupole time-of-flight MS/MS platforms. The method was testing on a fish matrix for recovery of palytoxin as well as marine corals for detection of natural mixtures of palytoxin analogues. The recovery of palytoxin was found to produce a linear response (R 2 of 0.95) when spiked into the fish matrix with a limit of quantitation of 2.5 ng/μL and recovery efficiency of 73% +/- 9%. The screening of corals revealed varying amount of palytoxin, and in one case, different palytoxin structural analogues. This demonstration illustrates the potential utility of this method for toxin extraction and detection.« less

  16. Method Development for Comprehensive Extraction and Analysis of Marine Toxins: Liquid-Liquid Extraction and Tandem Liquid Chromatography Separations Coupled to Electrospray Tandem Mass Spectrometry

    DOE PAGES

    Wunschel, David S.; Valenzuela, Blandina R.; Kaiser, Brooke L. Deatherage; ...

    2018-05-09

    A variety of toxins are produced by marine and freshwater microorganisms that present a threat to human health. These toxins have diverse chemical properties and specifically, a range of hydrophobicity. Methods for extraction and identification of these toxins are often geared toward specific classes of toxin depending on the sample type. There is a need for a general method of toxin extraction and identification for screening samples where the likely toxin content is not known a priori. Here, we have applied a general method for metabolite extraction to toxin containing samples. This method was coupled with a simple dual liquidmore » chromatography approach for separating a broad range of toxins. This liquid chromatography approach was coupled to triple quadrupole and quadrupole time-of-flight MS/MS platforms. The method was testing on a fish matrix for recovery of palytoxin as well as marine corals for detection of natural mixtures of palytoxin analogues. The recovery of palytoxin was found to produce a linear response (R 2 of 0.95) when spiked into the fish matrix with a limit of quantitation of 2.5 ng/μL and recovery efficiency of 73% +/- 9%. The screening of corals revealed varying amount of palytoxin, and in one case, different palytoxin structural analogues. This demonstration illustrates the potential utility of this method for toxin extraction and detection.« less

  17. Validated reversed phase LC method for quantitative analysis of polymethoxyflavones in citrus peel extracts.

    PubMed

    Wang, Zhenyu; Li, Shiming; Ferguson, Stephen; Goodnow, Robert; Ho, Chi-Tang

    2008-01-01

    Polymethoxyflavones (PMFs), which exist exclusively in the citrus genus, have biological activities including anti-inflammatory, anticarcinogenic, and antiatherogenic properties. A validated RPLC method was developed for quantitative analysis of six major PMFs, namely nobiletin, tangeretin, sinensetin, 5,6,7,4'-tetramethoxyflavone, 3,5,6,7,3',4'-hexamethoxyflavone, and 3,5,6,7,8,3',4'-heptamethoxyflavone. The polar embedded LC stationary phase was able to fully resolve the six analogues. The developed method was fully validated in terms of linearity, accuracy, precision, sensitivity, and system suitability. The LOD of the method was calculated as 0.15 microg/mL and the recovery rate was between 97.0 and 105.1%. This analytical method was successfully applied to quantify the individual PMFs in four commercially available citrus peel extracts (CPEs). Each extract shows significant difference in the PMF composition and concentration. This method may provide a simple, rapid, and reliable tool to help reveal the correlation between the bioactivity of the PMF extracts and the individual PMF content.

  18. An easy, rapid and inexpensive method to monitor tributyltin (TBT) toxicity in the laboratory.

    PubMed

    Cruz, Andreia; Moreira, Rafael; Mendo, Sónia

    2014-05-01

    Tributyltin (TBT) contamination remains a major problem worldwide. Many laboratories are committed to the development of remediation methodologies that could help reduce the negative impact of this compound in the environment. Furthermore, it is important to have at hand simple methodologies for evaluating TBT toxicity in the laboratory, besides the use of complex and costly analytical instrumentation. With that purpose, a method was adapted that is based on the inhibition of growth of an indicator strain, Micrococcus luteus ATCC 9341, under TBT. Different types of matrices, of TBT concentrations and sample treatments were tested. The results herein reported show that the bioassay method can be applied for both aqueous and soil samples and also for a high range of TBT concentrations (at least up to 500 μmol/L). Besides being cheap and easy to perform, it can be performed in any laboratory. Additionally, one possible application of the method to monitor TBT degradation is presented as an example.

  19. First beam measurements on the vessel for extraction and source plasma analyses (VESPA) at the Rutherford Appleton Laboratory (RAL)

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

    Lawrie, Scott R., E-mail: scott.lawrie@stfc.ac.uk; John Adams Institute for Accelerator Science, Department of Physics, University of Oxford; Faircloth, Daniel C.

    2015-04-08

    In order to facilitate the testing of advanced H{sup −} ion sources for the ISIS and Front End Test Stand (FETS) facilities at the Rutherford Appleton Laboratory (RAL), a Vessel for Extraction and Source Plasma Analyses (VESPA) has been constructed. This will perform the first detailed plasma measurements on the ISIS Penning-type H{sup −} ion source using emission spectroscopic techniques. In addition, the 30-year-old extraction optics are re-designed from the ground up in order to fully transport the beam. Using multiple beam and plasma diagnostics devices, the ultimate aim is improve H{sup −} production efficiency and subsequent transport for eithermore » long-term ISIS user operations or high power FETS requirements. The VESPA will also accommodate and test a new scaled-up Penning H{sup −} source design. This paper details the VESPA design, construction and commissioning, as well as initial beam and spectroscopy results.« less

  20. A method of directly extracting multiwave angle-domain common-image gathers

    NASA Astrophysics Data System (ADS)

    Han, Jianguang; Wang, Yun

    2017-10-01

    Angle-domain common-image gathers (ADCIGs) can provide an effective way for migration velocity analysis and amplitude versus angle analysis in oil-gas seismic exploration. On the basis of multi-component Gaussian beam prestack depth migration (GB-PSDM), an alternative method of directly extracting multiwave ADCIGs is presented in this paper. We first introduce multi-component GB-PSDM, where a wavefield separation is proceeded to obtain the separated PP- and PS-wave seismic records before migration imaging for multiwave seismic data. Then, the principle of extracting PP- and PS-ADCIGs using GB-PSDM is presented. The propagation angle can be obtained using the real-value travel time of Gaussian beam in the course of GB-PSDM, which can be used to calculate the incidence and reflection angles. Two kinds of ADCIGs can be extracted for the PS-wave, one of which is P-wave incidence ADCIGs and the other one is S-wave reflection ADCIGs. In this paper, we use the incident angle to plot the ADCIGs for both PP- and PS-waves. Finally, tests of synthetic examples show that the method introduced here is accurate and effective.

  1. Rapid extraction and assay of uranium from environmental surface samples

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

    Barrett, Christopher A.; Chouyyok, Wilaiwan; Speakman, Robert J.

    Extraction methods enabling faster removal and concentration of uranium compounds for improved trace and low-level assay are demonstrated for standard surface sampling material in support of nuclear safeguards efforts, health monitoring, and other nuclear analysis applications. A key problem with the existing surface sampling swipes is the requirement for complete digestion of sample and sampling matrix. This is a time-consuming and labour-intensive process that limits laboratory throughput, elevates costs, and increases background levels. Various extraction methods are explored for their potential to quickly and efficiently remove different chemical forms of uranium from standard surface sampling material. A combination of carbonatemore » and peroxide solutions is shown to give the most rapid and complete form of uranyl compound extraction and dissolution. This rapid extraction process is demonstrated to be compatible with standard inductive coupled plasma mass spectrometry methods for uranium isotopic assay as well as screening techniques such as x-ray fluorescence. The general approach described has application beyond uranium to other analytes of nuclear forensic interest (e.g., rare earth elements and plutonium) as well as heavy metals for environmental and industrial hygiene monitoring.« less

  2. Membrane contactor assisted water extraction system for separating hydrogen peroxide from a working solution, and method thereof

    DOEpatents

    Snyder, Seth W [Lincolnwood, IL; Lin, Yupo J [Naperville, IL; Hestekin', Jamie A [Fayetteville, AR; Henry, Michael P [Batavia, IL; Pujado, Peter [Kildeer, IL; Oroskar, Anil [Oak Brook, IL; Kulprathipanja, Santi [Inverness, IL; Randhava, Sarabjit [Evanston, IL

    2010-09-21

    The present invention relates to a membrane contactor assisted extraction system and method for extracting a single phase species from multi-phase working solutions. More specifically one preferred embodiment of the invention relates to a method and system for membrane contactor assisted water (MCAWE) extraction of hydrogen peroxide (H.sub.2O.sub.2) from a working solution.

  3. Comparison of methods for conducting marine and estuarine sediment porewater toxicity tests—extraction, storage, and handling techniques

    USGS Publications Warehouse

    Carr, R.S.; Chapman, D.C.

    1995-01-01

    A series of studies was conducted to compare different porewater extraction techniques and to evaluate the effects of sediment and porewater storage conditions on the toxicity of pore water, using assays with the sea urchin Arbacia punctulata. If care is taken in the selection of materials, several different porewater extraction techniques (pressurized squeezing, centrifugation, vacuum) yield samples with similar toxicity. Where the primary contaminants of concern are highly hydrophobic organic compounds, centrifugation is the method of choice for minimizing the loss of contaminants during the extraction procedure. No difference was found in the toxicity of pore water obtained with the Teflon® and polyvinyl chloride pressurized extraction devices. Different types of filters in the squeeze extraction devices apparently adsorbed soluble contaminants to varying degrees. The amount of fine suspended particulate material remaining in the pore water after the initial extraction varied among the methods. For most of the sediments tested, freezing and thawing did not affect the toxicity of porewater samples obtained by the pressurized squeeze extraction method. Pore water obtained by other methods (centrifugation, vacuum) and frozen without additional removal of suspended particulates by centrifugation may exhibit increased toxicity compared with the unfrozen sample.The toxicity of pore water extracted from refrigerated (4°C) sediments exhibited substantial short-term (days, weeks) changes. Similarly, sediment pore water extracted over time from a simulated amphipod solid-phase toxicity test changed substantially in toxicity. For the sediments tested, the direction and magnitude of change in toxicity of pore water extracted from both refrigerated and solid-phase test sediments was unpredictable.

  4. Soil sampling and extraction methods with possible application to pear thrips (Thysanoptera: Thripidae)

    Treesearch

    John E. Bater

    1991-01-01

    Techniques are described for the sampling and extraction of microarthropods from soil and the potential of these methods to extract the larval stages of the pear thrips, Taeniothrips inconsequens (Uzel), from soil cores taken in sugar maple stands. Also described is a design for an emergence trap that could be used to estimate adult thrips...

  5. Extraction of solubles from plant biomass for use as microbial growth stimulant and methods related thereto

    DOEpatents

    Lau, Ming Woei

    2015-12-08

    A method for producing a microbial growth stimulant (MGS) from a plant biomass is described. In one embodiment, an ammonium hydroxide solution is used to extract a solution of proteins and ammonia from the biomass. Some of the proteins and ammonia are separated from the extracted solution to provide the MGS solution. The removed ammonia can be recycled and the proteins are useful as animal feeds. In one embodiment, the method comprises extracting solubles from pretreated lignocellulosic biomass with a cellulase enzyme-producing growth medium (such T. reesei) in the presence of water and an aqueous extract.

  6. Influence of extractives on bonding properties of white and southern red oak

    Treesearch

    Mon-Lin Kuo; Daniel DiCarlo; Chung-Yun Hse

    1984-01-01

    White and southern red oak veneer were subjected to four methods of drying followed by five surface treatments. The four drying methods were mill drying at 350°F, laboratory drying at 350 and 212°F, and air drying. The five surface treatments were no treatment, surface scraping, soaking and dipping in 1% NaOH aqueous solution, and water extraction. Plywood panels were...

  7. A Manual of Simplified Laboratory Methods for Operators of Wastewater Treatment Facilities.

    ERIC Educational Resources Information Center

    Westerhold, Arnold F., Ed.; Bennett, Ernest C., Ed.

    This manual is designed to provide the small wastewater treatment plant operator, as well as the new or inexperienced operator, with simplified methods for laboratory analysis of water and wastewater. It is emphasized that this manual is not a replacement for standard methods but a guide for plants with insufficient equipment to perform analyses…

  8. A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples

    PubMed Central

    Oates, Lawrence G.; Read, Harry W.; Gutknecht, Jessica L. M.; Duncan, David S.; Balser, Teri B.; Jackson, Randall D.

    2017-01-01

    Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial communities, a relatively precise but effort-intensive technique to assay microbial community composition is phospholipid fatty acid (PLFA) analysis. PLFA was developed to analyze phospholipid biomarkers, which can be used as indicators of microbial biomass and the composition of broad functional groups of fungi and bacteria. It has commonly been used to compare soils under alternative plant communities, ecology, and management regimes. The PLFA method has been shown to be sensitive to detecting shifts in microbial community composition. An alternative method, fatty acid methyl ester extraction and analysis (MIDI-FA) was developed for rapid extraction of total lipids, without separation of the phospholipid fraction, from pure cultures as a microbial identification technique. This method is rapid but is less suited for soil samples because it lacks an initial step separating soil particles and begins instead with a saponification reaction that likely produces artifacts from the background organic matter in the soil. This article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples. The method combines the accuracy achieved through PLFA profiling by extracting and concentrating soil lipids as a first step, and a reduction in effort by saponifying the organic material extracted and processing with the MIDI-FA method as a second step. PMID:28745639

  9. Method for extracting metals from aqueous waste streams for long term storage

    DOEpatents

    Chaiko, D.J.

    1995-03-07

    A liquid-liquid extraction method for removing metals and hydrous metal colloids from waste streams is provided wherein said waste streams are contacted with a solvent system containing a water-in-oil microemulsion wherein the inverted micelles contain the extracted metal. A silicon alkoxide, either alone or in combination with other metal alkoxide compounds is added to the water-in-oil microemulsion, thereby allowing encapsulation of the extracted metal within a silicon oxide network. Lastly, the now-encapsulated metal is precipitated from the water-in-oil microemulsion phase to yield aggregates of metal-silicate particles having average individual particle sizes of approximately 40 nanometers. 2 figs.

  10. Method for extracting metals from aqueous waste streams for long term storage

    DOEpatents

    Chaiko, D.J.

    1993-01-01

    A liquid-liquid extraction method for removing metals and hydrous metal colloids from waste streams is provided wherein said waste streams are contacted with a solvent system containing a water-in-oil microemulsion wherein the inverted micelles contain the extracted metal. A silicon alkoxide, either alone or in combination with other metal alkoxide compounds is added to the water-in-oil microemulsion, thereby allowing encapsulation of the extracted metal within a silicon oxide network. Lastly, the now-encapsulated metal is precipitated from the water-in-oil microemulsion phase to yield aggregates of metal-silicate particles having average. individual particle sizes of approximately 40 manometers.

  11. Method for extracting metals from aqueous waste streams for long term storage

    DOEpatents

    Chaiko, David J.

    1995-01-01

    A liquid--liquid extraction method for removing metals and hydrous metal colloids from waste streams is provided wherein said waste streams are contacted with a solvent system containing a water-in-oil microemulsion wherein the inverted micelles contain the extracted metal. A silicon alkoxide, either alone or in combination with other metal alkoxide compounds is added to the water-in-oil microemulsion, thereby allowing encapsulation of the extracted metal within a silicon oxide network. Lastly, the now-encapsulated metal is precipitated from the water-in-oil microemulsion phase to yield aggregates of metal-silicate particles having average individual particle sizes of approximately 40 nanometers.

  12. A method for automatically extracting infectious disease-related primers and probes from the literature

    PubMed Central

    2010-01-01

    Background Primer and probe sequences are the main components of nucleic acid-based detection systems. Biologists use primers and probes for different tasks, some related to the diagnosis and prescription of infectious diseases. The biological literature is the main information source for empirically validated primer and probe sequences. Therefore, it is becoming increasingly important for researchers to navigate this important information. In this paper, we present a four-phase method for extracting and annotating primer/probe sequences from the literature. These phases are: (1) convert each document into a tree of paper sections, (2) detect the candidate sequences using a set of finite state machine-based recognizers, (3) refine problem sequences using a rule-based expert system, and (4) annotate the extracted sequences with their related organism/gene information. Results We tested our approach using a test set composed of 297 manuscripts. The extracted sequences and their organism/gene annotations were manually evaluated by a panel of molecular biologists. The results of the evaluation show that our approach is suitable for automatically extracting DNA sequences, achieving precision/recall rates of 97.98% and 95.77%, respectively. In addition, 76.66% of the detected sequences were correctly annotated with their organism name. The system also provided correct gene-related information for 46.18% of the sequences assigned a correct organism name. Conclusions We believe that the proposed method can facilitate routine tasks for biomedical researchers using molecular methods to diagnose and prescribe different infectious diseases. In addition, the proposed method can be expanded to detect and extract other biological sequences from the literature. The extracted information can also be used to readily update available primer/probe databases or to create new databases from scratch. PMID:20682041

  13. Puffed cereals with added chamomile - quantitative analysis of polyphenols and optimization of their extraction method.

    PubMed

    Blicharski, Tomasz; Oniszczuk, Anna; Olech, Marta; Oniszczuk, Tomasz; Wójtowicz, Agnieszka; Krawczyk, Wojciech; Nowak, Renata

    2017-05-11

    [b]Abstract Introduction[/b]. Functional food plays an important role in the prevention, management and treatment of chronic diseases. One of the most interesting techniques of functional food production is extrusion-cooking. Functional foods may include such items as puffed cereals, breads and beverages that are fortified with vitamins, some nutraceuticals and herbs. Due to its pharmacological activity, chamomile flowers are the most popular components added to functional food. Quantitative analysis of polyphenolic antioxidants, as well as comparison of various methods for the extraction of phenolic compounds from corn puffed cereals, puffed cereals with an addition of chamomile (3, 5, 10 and 20%) and from [i]Chamomillae anthodium. [/i] [b]Materials and Methods[/b]. Two modern extraction methods - ultrasound assisted extraction (UAE) at 40 °C and 60 °C, as well as accelerated solvent extraction (ASE) at 100 °C and 120 °C were used for the isolation of polyphenols from functional food. Analysis of flavonoids and phenolic acids was carried out using reversed-phase high-performance liquid chromatography and electrospray ionization mass spectrometry (LC-ESI-MS/MS). [b]Results and Conclusions[/b]. For most of the analyzed compounds, the highest yields were obtained by ultrasound assisted extraction. The highest temperature during the ultrasonification process (60 °C) increased the efficiency of extraction, without degradation of polyphenols. UAE easily arrives at extraction equilibrium and therefore permits shorter periods of time, reducing the energy input. Furthermore, UAE meets the requirements of 'Green Chemistry'.

  14. The continued value of disk diffusion for assessing antimicrobial susceptibility in clinical laboratories: report from the Clinical and Laboratory Standards Institute Methods Development and Standardization Working Group.

    PubMed

    Humphries, Romney M; Kircher, Susan; Ferrell, Andrea; Krause, Kevin M; Malherbe, Rianna; Hsiung, Andre; Burnham, C A

    2018-05-09

    Expedited pathways to antimicrobial agent approval by the United States Food and Drug Administration (FDA) have led to increased delays between drug approval and the availability of FDA-cleared antimicrobial susceptibility testing (AST) devices. Antimicrobial disks for use with disk diffusion testing are among the first AST devices available to clinical laboratories. However, many laboratories are reluctant to implement a disk diffusion method for a variety of reasons, including dwindling proficiency with this method, interruptions to laboratory workflow, uncertainty surrounding the quality and reliability of a disk diffusion test, and perceived need to report an MIC to clinicians. This mini-review provides a report from the Clinical and Laboratory Standards Institute Working Group on Methods Development and Standardization on the current standards and clinical utility of disk diffusion testing. Copyright © 2018 American Society for Microbiology.

  15. Comparability between various field and laboratory wood-stove emission-measurement methods

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

    McCrillis, R.C.; Jaasma, D.R.

    1991-01-01

    The paper compares various field and laboratory woodstove emission measurement methods. In 1988, the U.S. EPA promulgated performance standards for residential wood heaters (woodstoves). Over the past several years, a number of field studies have been undertaken to determine the actual level of emission reduction achieved by new technology woodstoves in everyday use. The studies have required the development and use of particulate and gaseous emission sampling equipment compatible with operation in private homes. Since woodstoves are tested for certification in the laboratory using EPA Methods 5G and 5H, it is of interest to determine the correlation between these regulatorymore » methods and the inhouse equipment. Two inhouse sampling systems have been used most widely: one is an intermittent, pump-driven particulate sampler that collects particulate and condensible organics on a filter and organic adsorbent resin; and the other uses an evacuated cylinder as the motive force and particulate and condensible organics are collected in a condenser and dual filter. Both samplers can operate unattended for 1-week periods. A large number of tests have been run comparing Methods 5G and 5H to both samplers. The paper presents these comparison data and determines the relationships between regulations and field samplers.« less

  16. Method for evaluation of laboratory craters using crater detection algorithm for digital topography data

    NASA Astrophysics Data System (ADS)

    Salamunićcar, Goran; Vinković, Dejan; Lončarić, Sven; Vučina, Damir; Pehnec, Igor; Vojković, Marin; Gomerčić, Mladen; Hercigonja, Tomislav

    In our previous work the following has been done: (1) the crater detection algorithm (CDA) based on digital elevation model (DEM) has been developed and the GT-115225 catalog has been assembled [GRS, 48 (5), in press, doi:10.1109/TGRS.2009.2037750]; and (2) the results of comparison between explosion-induced laboratory craters in stone powder surfaces and GT-115225 have been presented using depth/diameter measurements [41stLPSC, Abstract #1428]. The next step achievable using the available technology is to create 3D scans of such labo-ratory craters, in order to compare different properties with simple Martian craters. In this work, we propose a formal method for evaluation of laboratory craters, in order to provide objective, measurable and reproducible estimation of the level of achieved similarity between these laboratory and real impact craters. In the first step, the section of MOLA data for Mars (or SELENE LALT for Moon) is replaced with one or several 3D-scans of laboratory craters. Once embedment was done, the CDA can be used to find out whether this laboratory crater is similar enough to real craters, as to be recognized as a crater by the CDA. The CDA evaluation using ROC' curve represents how true detection rate (TDR=TP/(TP+FN)=TP/GT) depends on the false detection rate (FDR=FP/(TP+FP)). Using this curve, it is now possible to define the measure of similarity between laboratory and real impact craters, as TDR or FDR value, or as a distance from the bottom-right origin of the ROC' curve. With such an approach, the reproducible (formally described) method for evaluation of laboratory craters is provided.

  17. Effects of different extraction methods on total phenolic content and antioxidant activity in soybean cultivars

    NASA Astrophysics Data System (ADS)

    Yusnawan, E.

    2018-01-01

    Soybean secondary metabolites particularly phenolic compounds act as chemical defence against biotic stress such as pathogen infection. Functional properties of these compounds have also been investigated. This study aimed to determine the effects of particle size and extraction methods on total flavonoid, phenolic contents as well as antioxidant activity in soybean seeds. This study also investigated the total phenolic contents and antioxidant activity of Indonesian soybean cultivars using the optimized extraction method. Soybean flour of ≤ 177 μm as many as 0.5 g was selected for extraction with 50% acetone for estimation of total phenolic and flavonoid contents and with 80% ethanol for antioxidant activity. Treatments of twice extraction either shaking followed by maceration or ultrasound-assisted extraction followed by maceration could be used to extract the secondary metabolite contents in soybean seeds. Flavonoid, phenolic contents and antioxidant activity of twenty soybean cultivars ranged from 0.23 to 0.44 mg CE/g, from 3.70 to 5.22 mg GAE/g, and from 4.97 to 9.04 µmol TE/g, respectively. A simple extraction with small amount of soybean flour such as investigated in this present study is effective to extract secondary metabolites especially when the availability of samples is limited such as breeding materials or soybean germplasm.

  18. Stable Extraction of Threshold Voltage Using Transconductance Change Method for CMOS Modeling, Simulation and Characterization

    NASA Astrophysics Data System (ADS)

    Choi, Woo Young; Woo, Dong-Soo; Choi, Byung Yong; Lee, Jong Duk; Park, Byung-Gook

    2004-04-01

    We proposed a stable extraction algorithm for threshold voltage using transconductance change method by optimizing node interval. With the algorithm, noise-free gm2 (=dgm/dVGS) profiles can be extracted within one-percent error, which leads to more physically-meaningful threshold voltage calculation by the transconductance change method. The extracted threshold voltage predicts the gate-to-source voltage at which the surface potential is within kT/q of φs=2φf+VSB. Our algorithm makes the transconductance change method more practical by overcoming noise problem. This threshold voltage extraction algorithm yields the threshold roll-off behavior of nanoscale metal oxide semiconductor field effect transistor (MOSFETs) accurately and makes it possible to calculate the surface potential φs at any other point on the drain-to-source current (IDS) versus gate-to-source voltage (VGS) curve. It will provide us with a useful analysis tool in the field of device modeling, simulation and characterization.

  19. A novel method to augment extraction of mangiferin by application of microwave on three phase partitioning.

    PubMed

    Kulkarni, Vrushali M; Rathod, Virendra K

    2015-06-01

    This work reports a novel approach where three phase partitioning (TPP) was combined with microwave for extraction of mangiferin from leaves of Mangifera indica . Soxhlet extraction was used as reference method, which yielded 57 mg/g in 5 h. Under optimal conditions such as microwave irradiation time 5 min, ammonium sulphate concentration 40% w/v, power 272 W, solute to solvent ratio 1:20, slurry to t -butanol ratio 1:1, soaking time 5 min and duty cycle 50%, the mangiferin yield obtained was 54 mg/g by microwave assisted three phase partitioning extraction (MTPP). Thus extraction method developed resulted into higher extraction yield in a shorter span, thereby making it an interesting alternative prior to down-stream processing.

  20. Comparison of four extraction/methylation analytical methods to measure fatty acid composition by gas chromatography in meat.

    PubMed

    Juárez, M; Polvillo, O; Contò, M; Ficco, A; Ballico, S; Failla, S

    2008-05-09

    Four different extraction-derivatization methods commonly used for fatty acid analysis in meat (in situ or one-step method, saponification method, classic method and a combination of classic extraction and saponification derivatization) were tested. The in situ method had low recovery and variation. The saponification method showed the best balance between recovery, precision, repeatability and reproducibility. The classic method had high recovery and acceptable variation values, except for the polyunsaturated fatty acids, showing higher variation than the former methods. The combination of extraction and methylation steps had great recovery values, but the precision, repeatability and reproducibility were not acceptable. Therefore the saponification method would be more convenient for polyunsaturated fatty acid analysis, whereas the in situ method would be an alternative for fast analysis. However the classic method would be the method of choice for the determination of the different lipid classes.

  1. Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory : determination of polycyclic aromatic hydrocarbon compounds in sediment by gas chromatography/mass spectrometry

    USGS Publications Warehouse

    Olson, Mary C.; Iverson, Jana L.; Furlong, Edward T.; Schroeder, Michael P.

    2004-01-01

    A method for the determination of 28 polycyclic aromatic hydrocarbons (PAHs) and 25 alkylated PAH homolog groups in sediment samples is described. The compounds are extracted from sediment by solvent extraction, followed by partial isolation using high-performance gel permeation chromatography. The compounds are identified and uantitated using capillary-column gas chromatography/mass spectrometry. The report presents performance data for full-scan ion monitoring. Method detection limits in laboratory reagent matrix samples range from 1.3 to 5.1 micrograms per kilogram for the 28 PAHs. The 25 groups of alkylated PAHs are homologs of five groups of isomeric parent PAHs. Because of the lack of authentic standards, these homologs are reported semiquantitatively using a response factor from a parent PAH or a specific alkylated PAH. Precision data for the alkylated PAH homologs are presented using two different standard reference manuals produced by the National Institute of Standards and Technology: SRM 1941b and SRM 1944. The percent relative standard deviations for identified alkylated PAH homolog groups ranged from 1.55 to 6.98 for SRM 1941b and from 6.11 to 12.0 for SRM 1944. Homolog group concentrations reported under this method include the concentrations of individually identified compounds that are members of the group. Organochlorine (OC) pesticides--including toxaphene, polychlorinated biphenyls (PCBs), and organophosphate (OP) pesticides--can be isolated simultaneously using this method. In brief, sediment samples are centrifuged to remove excess water and extracted overnight with dichloromethan (95 percent) and methanol (5 percent). The extract is concentrated and then filtered through a 0.2-micrometer polytetrafluoroethylene syringe filter. The PAH fraction is isolated by quantitatively injecting an aliquot of sample onto two polystyrene-divinylbenzene gel-permeation chromatographic columns connected in series. The compounds are eluted with dichloromethane

  2. Comparison of Boiling and Robotics Automation Method in DNA Extraction for Metagenomic Sequencing of Human Oral Microbes.

    PubMed

    Yamagishi, Junya; Sato, Yukuto; Shinozaki, Natsuko; Ye, Bin; Tsuboi, Akito; Nagasaki, Masao; Yamashita, Riu

    2016-01-01

    The rapid improvement of next-generation sequencing performance now enables us to analyze huge sample sets with more than ten thousand specimens. However, DNA extraction can still be a limiting step in such metagenomic approaches. In this study, we analyzed human oral microbes to compare the performance of three DNA extraction methods: PowerSoil (a method widely used in this field), QIAsymphony (a robotics method), and a simple boiling method. Dental plaque was initially collected from three volunteers in the pilot study and then expanded to 12 volunteers in the follow-up study. Bacterial flora was estimated by sequencing the V4 region of 16S rRNA following species-level profiling. Our results indicate that the efficiency of PowerSoil and QIAsymphony was comparable to the boiling method. Therefore, the boiling method may be a promising alternative because of its simplicity, cost effectiveness, and short handling time. Moreover, this method was reliable for estimating bacterial species and could be used in the future to examine the correlation between oral flora and health status. Despite this, differences in the efficiency of DNA extraction for various bacterial species were observed among the three methods. Based on these findings, there is no "gold standard" for DNA extraction. In future, we suggest that the DNA extraction method should be selected on a case-by-case basis considering the aims and specimens of the study.

  3. Critical points of DNA quantification by real-time PCR – effects of DNA extraction method and sample matrix on quantification of genetically modified organisms

    PubMed Central

    Cankar, Katarina; Štebih, Dejan; Dreo, Tanja; Žel, Jana; Gruden, Kristina

    2006-01-01

    Background Real-time PCR is the technique of choice for nucleic acid quantification. In the field of detection of genetically modified organisms (GMOs) quantification of biotech products may be required to fulfil legislative requirements. However, successful quantification depends crucially on the quality of the sample DNA analyzed. Methods for GMO detection are generally validated on certified reference materials that are in the form of powdered grain material, while detection in routine laboratories must be performed on a wide variety of sample matrixes. Due to food processing, the DNA in sample matrixes can be present in low amounts and also degraded. In addition, molecules of plant origin or from other sources that affect PCR amplification of samples will influence the reliability of the quantification. Further, the wide variety of sample matrixes presents a challenge for detection laboratories. The extraction method must ensure high yield and quality of the DNA obtained and must be carefully selected, since even components of DNA extraction solutions can influence PCR reactions. GMO quantification is based on a standard curve, therefore similarity of PCR efficiency for the sample and standard reference material is a prerequisite for exact quantification. Little information on the performance of real-time PCR on samples of different matrixes is available. Results Five commonly used DNA extraction techniques were compared and their suitability for quantitative analysis was assessed. The effect of sample matrix on nucleic acid quantification was assessed by comparing 4 maize and 4 soybean matrixes. In addition 205 maize and soybean samples from routine analysis were analyzed for PCR efficiency to assess variability of PCR performance within each sample matrix. Together with the amount of DNA needed for reliable quantification, PCR efficiency is the crucial parameter determining the reliability of quantitative results, therefore it was chosen as the primary

  4. Two-step extraction method for lead isotope fractionation to reveal anthropogenic lead pollution.

    PubMed

    Katahira, Kenshi; Moriwaki, Hiroshi; Kamura, Kazuo; Yamazaki, Hideo

    2018-05-28

    This study developed the 2-step extraction method which eluted the Pb adsorbing on the surface of sediments in the first solution by aqua regia and extracted the Pb absorbed inside particles into the second solution by mixed acid of nitric acid, hydrofluoric acid and hydrogen peroxide solution. We applied the method to sediments in the enclosed water area and found out that the isotope ratios of Pb in the second solution represented those of natural origin. This advantage of the method makes it possible to distinguish the Pb between natural origin and anthropogenic source on the basis of the isotope ratios. The results showed that the method was useful to discuss the Pb sources and that anthropogenic Pb in the sediment samples analysed was mainly derived from China because of transboundary air pollution.

  5. Laboratory Evaluation of Acoustic Backscatter and LISST Methods for Measurements of Suspended Sediments

    PubMed Central

    Meral, Ramazan

    2008-01-01

    The limitation of traditional sampling method to provide detailed spatial and temporal profiles of suspended sediment concentration has led to an interest in alternative devices and methods based on scattering of underwater sound and light. In the present work, acoustic backscatter and LISST (the Laser In Situ Scattering Transmissometry) devices, and methodologies were given. Besides a laboratory study was conducted to compare pumping methods for different sediment radiuses at the same concentration. The glass spheres (ballotini) of three different radiuses of 115, 137 and 163 μm were used to obtain suspension in the sediment tower at laboratory. A quite good agreement was obtained between these methods and pumping results with the range at 60.6-94.2% for sediment concentration and 91.3-100% for radius measurements. These results and the other studies show that these methods have potential for research tools for sediment studies. In addition further studies are needed to determine the ability of these methods for sediment measurement under different water and sediment material conditions. PMID:27879747

  6. Laboratory Production of Lemon Liqueur (Limoncello) by Conventional Maceration and a Two-Syringe System to Illustrate Rapid Solid-Liquid Dynamic Extraction

    ERIC Educational Resources Information Center

    Naviglio, Daniele; Montesano, Domenico; Gallo, Monica

    2015-01-01

    Two experimental techniques of solid-liquid extraction are compared relating to the lab-scale production of lemon liqueur, most commonly named "limoncello"; the first is the official method of maceration for the solid-liquid extraction of analytes and is widely used to extract active ingredients from a great variety of natural products;…

  7. Microgravimetric Analysis Method for Activation-Energy Extraction from Trace-Amount Molecule Adsorption.

    PubMed

    Xu, Pengcheng; Yu, Haitao; Li, Xinxin

    2016-05-03

    Activation-energy (Ea) value for trace-amount adsorption of gas molecules on material is rapidly and inexpensively obtained, for the first time, from a microgravimetric analysis experiment. With the material loaded, a resonant microcantilever is used to record in real time the adsorption process at two temperatures. The kinetic parameter Ea is thereby extracted by solving the Arrhenius equation. As an example, two CO2 capture nanomaterials are examined by the Ea extracting method for evaluation/optimization and, thereby, demonstrating the applicability of the microgravimetric analysis method. The achievement helps to solve the absence in rapid quantitative characterization of sorption kinetics and opens a new route to investigate molecule adsorption processes and materials.

  8. DNA Extraction from Soils: Old Bias for New Microbial Diversity Analysis Methods

    PubMed Central

    Martin-Laurent, F.; Philippot, L.; Hallet, S.; Chaussod, R.; Germon, J. C.; Soulas, G.; Catroux, G.

    2001-01-01

    The impact of three different soil DNA extraction methods on bacterial diversity was evaluated using PCR-based 16S ribosomal DNA analysis. DNA extracted directly from three soils showing contrasting physicochemical properties was subjected to amplified ribosomal DNA restriction analysis and ribosomal intergenic spacer analysis (RISA). The obtained RISA patterns revealed clearly that both the phylotype abundance and the composition of the indigenous bacterial community are dependent on the DNA recovery method used. In addition, this effect was also shown in the context of an experimental study aiming to estimate the impact on soil biodiversity of the application of farmyard manure or sewage sludge onto a monoculture of maize for 15 years. PMID:11319122

  9. Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes.

    PubMed

    Mikaeili, F; Kia, E B; Sharbatkhori, M; Sharifdini, M; Jalalizand, N; Heidari, Z; Zarei, Z; Stensvold, C R; Mirhendi, H

    2013-06-01

    Six simple methods for extraction of ribosomal and mitochondrial DNA from Toxocara canis, Toxocara cati and Toxascaris leonina were compared by evaluating the presence, appearance and intensity of PCR products visualized on agarose gels and amplified from DNA extracted by each of the methods. For each species, two isolates were obtained from the intestines of their respective hosts: T. canis and T. leonina from dogs, and T. cati from cats. For all isolates, total DNA was extracted using six different methods, including grinding, boiling, crushing, beating, freeze-thawing and the use of a commercial kit. To evaluate the efficacy of each method, the internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit 1 (cox1) gene were chosen as representative markers for ribosomal and mitochondrial DNA, respectively. Among the six DNA extraction methods, the beating method was the most cost effective for all three species, followed by the commercial kit. Both methods produced high intensity bands on agarose gels and were characterized by no or minimal smear formation, depending on gene target; however, beating was less expensive. We therefore recommend the beating method for studies where costs need to be kept at low levels. Copyright © 2013 Elsevier Inc. All rights reserved.

  10. Evaluation of DNA extraction methods and their clinical application for direct detection of causative bacteria in continuous ambulatory peritoneal dialysis culture fluids from patients with peritonitis by using broad-range PCR.

    PubMed

    Kim, Si Hyun; Jeong, Haeng Soon; Kim, Yeong Hoon; Song, Sae Am; Lee, Ja Young; Oh, Seung Hwan; Kim, Hye Ran; Lee, Jeong Nyeo; Kho, Weon-Gyu; Shin, Jeong Hwan

    2012-03-01

    The aims of this study were to compare several DNA extraction methods and 16S rDNA primers and to evaluate the clinical utility of broad-range PCR in continuous ambulatory peritoneal dialysis (CAPD) culture fluids. Six type strains were used as model organisms in dilutions from 10(8) to 10(0) colony-forming units (CFU)/mL for the evaluation of 5 DNA extraction methods and 5 PCR primer pairs. Broad-range PCR was applied to 100 CAPD culture fluids, and the results were compared with conventional culture results. There were some differences between the various DNA extraction methods and primer sets with regard to the detection limits. The InstaGene Matrix (Bio-Rad Laboratories, USA) and Exgene Clinic SV kits (GeneAll Biotechnology Co. Ltd, Korea) seem to have higher sensitivities than the others. The results of broad-range PCR were concordant with the results from culture in 97% of all cases (97/100). Two culture-positive cases that were broad-range PCR-negative were identified as Candida albicans, and 1 PCR-positive but culture-negative sample was identified as Bacillus circulans by sequencing. Two samples among 54 broad-range PCR-positive products could not be sequenced. There were differences in the analytical sensitivity of various DNA extraction methods and primers for broad-range PCR. The broad-range PCR assay can be used to detect bacterial pathogens in CAPD culture fluid as a supplement to culture methods.

  11. Ferrous iron phosphorus in sediments: development of a quantification method through 2,2'-bipyridine extraction.

    PubMed

    Li, Qingman; Wang, Xingxiang; Bartlett, Rebecca; Pinay, Gilles; Kan, Dan; Zhang, Wen; Sun, Jingxian

    2012-11-01

    The role of ferrous iron in the phosphorus cycle of an aquatic ecosystem is poorly understood because of a lack of suitable methods to quantitatively evaluate ferrous iron phosphorus (FIP) phases. Using sediments sampled from Fubao Bay of Dianchi Lake in China, a novel extraction method for FIP using 2,2'-bipyridine was explored. Total phosphorus and iron in the sediments ranged from 1.0 to 5.0 mg/g (dry weight) and 28.5 to 90.6 mg/g, respectively. Organic content (as indicated by loss on ignition or LOI) and iron(II) ranged from 3.1 to 27.0% and 26.5 to 64.9 mg/g, respectively. The dissolution dynamics of FIP extraction with a low solid/liquid ratio (1:25) indicated that a single application of 0.2% 2,2'-bipyridine extracted both iron(II) (Fe(II)) and phosphorus (as PO4(3-)) in sediments with different organic contents with low efficiency. The extraction efficiency of Fe(II) was improved by alteration of the solid/liquid ratio, but the effect was limited. However, addition of a 1:1000 solid/liquid ratio of 0.5 M potassium chloride to a 0.2% 2,2'-bipyridine solution significantly accelerated extraction of FIP with the release of Fe(II) and phosphorus toward equilibrium at approximately 150 hours. Further investigation demonstrated that 2,2'-bipyridine exhibited a higher selectivity in distinguishing FIP from phosphorus bound to ferric (Fe(III)) oxides or precipitated by calcium (Ca2+). Air-drying sediments significantly decreased the amount of extracted FIP, which indicates that fresh, wet sediment should be used in this type of FIP extraction. Based on experimental results using the proposed extraction protocol, (1) FIP in sediments of Fubao Bay had a predominant status in the lake sediment and accounted for 23.4 to 39.8% of total phosphorus, and (2) Fe(II)(FIP) released in the extraction is directly proportional to phosphorus(FIP) (Fe(II)(FIP) = 2.84 x P(FIP) + 0.0007; R2 = 0.97) with an average molar ratio of Fe(II)(FIP)/P(FIP) of 2.7. This study shows that FIP

  12. An ensemble method for extracting adverse drug events from social media.

    PubMed

    Liu, Jing; Zhao, Songzheng; Zhang, Xiaodi

    2016-06-01

    Because adverse drug events (ADEs) are a serious health problem and a leading cause of death, it is of vital importance to identify them correctly and in a timely manner. With the development of Web 2.0, social media has become a large data source for information on ADEs. The objective of this study is to develop a relation extraction system that uses natural language processing techniques to effectively distinguish between ADEs and non-ADEs in informal text on social media. We develop a feature-based approach that utilizes various lexical, syntactic, and semantic features. Information-gain-based feature selection is performed to address high-dimensional features. Then, we evaluate the effectiveness of four well-known kernel-based approaches (i.e., subset tree kernel, tree kernel, shortest dependency path kernel, and all-paths graph kernel) and several ensembles that are generated by adopting different combination methods (i.e., majority voting, weighted averaging, and stacked generalization). All of the approaches are tested using three data sets: two health-related discussion forums and one general social networking site (i.e., Twitter). When investigating the contribution of each feature subset, the feature-based approach attains the best area under the receiver operating characteristics curve (AUC) values, which are 78.6%, 72.2%, and 79.2% on the three data sets. When individual methods are used, we attain the best AUC values of 82.1%, 73.2%, and 77.0% using the subset tree kernel, shortest dependency path kernel, and feature-based approach on the three data sets, respectively. When using classifier ensembles, we achieve the best AUC values of 84.5%, 77.3%, and 84.5% on the three data sets, outperforming the baselines. Our experimental results indicate that ADE extraction from social media can benefit from feature selection. With respect to the effectiveness of different feature subsets, lexical features and semantic features can enhance the ADE extraction

  13. A study of an aroma extraction method and evaluation of the aroma extract contribution to the palatability and reinforcement effect of dried bonito using mice.

    PubMed

    Amitsuka, Takahiko; Okamura, Maya; Shiibashi, Hiroko; Yamamoto, Naoto; Saito, Tsukasa; Nammoku, Takashi; Tsuzuki, Satoshi; Inoue, Kazuo; Fushiki, Tohru

    2014-01-01

    Japanese cuisine has provided satisfying meals by fully utilizing the characteristic aroma and taste of katsuodashi (dried bonito broth), though it is not rich in sugars or fats. Katsuodashi is a very basic and indispensable element in Japanese cuisine, and is a hot water extract of katsuobushi (dried bonito). It has been reported that a dextrin solution containing natural dried bonito broth has a significant reinforcement effect, and has been suggested that the olfactory stimulation is important for the reinforcement effect. We examined various source materials for broth and identified an optimal method of aroma extraction by two-bottle choice and conditioned place preference tests in mice. By two-bottle choice tests, a solution containing arabushi (a type of katsuobushi) aroma extract obtained by a supercritical CO2 extraction method showed a significantly high preference. The conditioned place preference test showed the dashi-taste solution with arabushi supercritical CO2 extract had a reinforcement effect. Our results suggest that the arabushi extract obtained by supercritical CO2 extraction contains components responsible for preference and reinforcement effects in mice; it could become conducive to making Japanese cuisine more satisfying and palatable.

  14. Fusion method of SAR and optical images for urban object extraction

    NASA Astrophysics Data System (ADS)

    Jia, Yonghong; Blum, Rick S.; Li, Fangfang

    2007-11-01

    A new image fusion method of SAR, Panchromatic (Pan) and multispectral (MS) data is proposed. First of all, SAR texture is extracted by ratioing the despeckled SAR image to its low pass approximation, and is used to modulate high pass details extracted from the available Pan image by means of the á trous wavelet decomposition. Then, high pass details modulated with the texture is applied to obtain the fusion product by HPFM (High pass Filter-based Modulation) fusion method. A set of image data including co-registered Landsat TM, ENVISAT SAR and SPOT Pan is used for the experiment. The results demonstrate accurate spectral preservation on vegetated regions, bare soil, and also on textured areas (buildings and road network) where SAR texture information enhances the fusion product, and the proposed approach is effective for image interpret and classification.

  15. Laboratory procedures to generate viral metagenomes.

    PubMed

    Thurber, Rebecca V; Haynes, Matthew; Breitbart, Mya; Wegley, Linda; Rohwer, Forest

    2009-01-01

    This collection of laboratory protocols describes the steps to collect viruses from various samples with the specific aim of generating viral metagenome sequence libraries (viromes). Viral metagenomics, the study of uncultured viral nucleic acid sequences from different biomes, relies on several concentration, purification, extraction, sequencing and heuristic bioinformatic methods. No single technique can provide an all-inclusive approach, and therefore the protocols presented here will be discussed in terms of hypothetical projects. However, care must be taken to individualize each step depending on the source and type of viral-particles. This protocol is a description of the processes we have successfully used to: (i) concentrate viral particles from various types of samples, (ii) eliminate contaminating cells and free nucleic acids and (iii) extract, amplify and purify viral nucleic acids. Overall, a sample can be processed to isolate viral nucleic acids suitable for high-throughput sequencing in approximately 1 week.

  16. General method for extracting the quantum efficiency of dispersive qubit readout in circuit QED

    NASA Astrophysics Data System (ADS)

    Bultink, C. C.; Tarasinski, B.; Haandbæk, N.; Poletto, S.; Haider, N.; Michalak, D. J.; Bruno, A.; DiCarlo, L.

    2018-02-01

    We present and demonstrate a general three-step method for extracting the quantum efficiency of dispersive qubit readout in circuit QED. We use active depletion of post-measurement photons and optimal integration weight functions on two quadratures to maximize the signal-to-noise ratio of the non-steady-state homodyne measurement. We derive analytically and demonstrate experimentally that the method robustly extracts the quantum efficiency for arbitrary readout conditions in the linear regime. We use the proven method to optimally bias a Josephson traveling-wave parametric amplifier and to quantify different noise contributions in the readout amplification chain.

  17. Multicenter evaluation of the Sepsityper™ extraction kit and MALDI-TOF MS for direct identification of positive blood culture isolates using the BD BACTEC™ FX and VersaTREK(®) diagnostic blood culture systems.

    PubMed

    Schieffer, K M; Tan, K E; Stamper, P D; Somogyi, A; Andrea, S B; Wakefield, T; Romagnoli, M; Chapin, K C; Wolk, D M; Carroll, K C

    2014-04-01

    (i) Evaluation of delayed time to blood culture extraction by the Sepsityper kit and impact of shipping pellets off-site for MALDI-TOF MS analysis. (ii) Comparison of Sepsityper and laboratory-developed extraction methods from a literature review. Using two blood culture systems (BD BACTEC and VersaTREK), we extracted 411 positive blood cultures using the Sepsityper kit to mimic a potential protocol for institutions without a MALDI-TOF MS. Extracted pellets were shipped and analysed on the Bruker UltraflexIII. Successful extraction of 358 (87·1%) samples was determined by the presence of detectable proteins. MALDI-TOF MS correctly identified 332 (80·8%) samples. Delayed time to extraction did not affect Sepsityper extraction or MALDI-TOF MS accuracy. The extracted pellets remain stable and provide accurate results by MALDI-TOF MS when shipped at room temperature to off-site reference laboratories. This is the first study to show that institutions without a MALDI-TOF MS can take advantage of this innovative technology by shipping a volume of blood to an off-site laboratory for extraction and MALDI-TOF MS analysis. We also performed a literature review to compare various extraction methods. © 2014 The Society for Applied Microbiology.

  18. Research on Methods of High Coherent Target Extraction in Urban Area Based on Psinsar Technology

    NASA Astrophysics Data System (ADS)

    Li, N.; Wu, J.

    2018-04-01

    PSInSAR technology has been widely applied in ground deformation monitoring. Accurate identification of Persistent Scatterers (PS) is key to the success of PSInSAR data processing. In this paper, the theoretic models and specific algorithms of PS point extraction methods are summarized and the characteristics and applicable conditions of each method, such as Coherence Coefficient Threshold method, Amplitude Threshold method, Dispersion of Amplitude method, Dispersion of Intensity method, are analyzed. Based on the merits and demerits of different methods, an improved method for PS point extraction in urban area is proposed, that uses simultaneously backscattering characteristic, amplitude and phase stability to find PS point in all pixels. Shanghai city is chosen as an example area for checking the improvements of the new method. The results show that the PS points extracted by the new method have high quality, high stability and meet the strong scattering characteristics. Based on these high quality PS points, the deformation rate along the line-of-sight (LOS) in the central urban area of Shanghai is obtained by using 35 COSMO-SkyMed X-band SAR images acquired from 2008 to 2010 and it varies from -14.6 mm/year to 4.9 mm/year. There is a large sedimentation funnel in the cross boundary of Hongkou and Yangpu district with a maximum sedimentation rate of more than 14 mm per year. The obtained ground subsidence rates are also compared with the result of spirit leveling and show good consistent. Our new method for PS point extraction is more reasonable, and can improve the accuracy of the obtained deformation results.

  19. Nuclear deformation in the laboratory frame

    NASA Astrophysics Data System (ADS)

    Gilbreth, C. N.; Alhassid, Y.; Bertsch, G. F.

    2018-01-01

    We develop a formalism for calculating the distribution of the axial quadrupole operator in the laboratory frame within the rotationally invariant framework of the configuration-interaction shell model. The calculation is carried out using a finite-temperature auxiliary-field quantum Monte Carlo method. We apply this formalism to isotope chains of even-mass samarium and neodymium nuclei and show that the quadrupole distribution provides a model-independent signature of nuclear deformation. Two technical advances are described that greatly facilitate the calculations. The first is to exploit the rotational invariance of the underlying Hamiltonian to reduce the statistical fluctuations in the Monte Carlo calculations. The second is to determine quadruple invariants from the distribution of the axial quadrupole operator in the laboratory frame. This allows us to extract effective values of the intrinsic quadrupole shape parameters without invoking an intrinsic frame or a mean-field approximation.

  20. A method for developing outcome measures in the clinical laboratory.

    PubMed

    Jones, J

    1996-01-01

    Measuring and reporting outcomes in health care is becoming more important for quality assessment, utilization assessment, accreditation standards, and negotiating contracts in managed care. How does one develop an outcome measure for the laboratory to assess the value of the services? A method is described which outlines seven steps in developing outcome measures for a laboratory service or process. These steps include the following: 1. Identify the process or service to be monitored for performance and outcome assessment. 2. If necessary, form an multidisciplinary team of laboratory staff, other department staff, physicians, and pathologists. 3. State the purpose of the test or service including a review of published data for the clinical pathological correlation. 4. Prepare a process cause and effect diagram including steps critical to the outcome. 5. Identify key process variables that contribute to positive or negative outcomes. 6. Identify outcome measures that are not process measures. 7. Develop an operational definition, identify data sources, and collect data. Examples, including a process cause and effect diagram, process variables, and outcome measures, are given using the Therapeutic Drug Monitoring service (TDM). A summary of conclusions and precautions for outcome measurement is then provided.