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Sample records for acid extraction protocol

  1. Fluorometric Quantification of Polyphosphate in Environmental Plankton Samples: Extraction Protocols, Matrix Effects, and Nucleic Acid Interference

    PubMed Central

    Martin, Patrick

    2013-01-01

    Polyphosphate (polyP) is a ubiquitous biochemical with many cellular functions and comprises an important environmental phosphorus pool. However, methodological challenges have hampered routine quantification of polyP in environmental samples. We tested 15 protocols to extract inorganic polyphosphate from natural marine samples and cultured cyanobacteria for fluorometric quantification with 4′,6-diamidino-2-phenylindole (DAPI) without prior purification. A combination of brief boiling and digestion with proteinase K was superior to all other protocols, including other enzymatic digestions and neutral or alkaline leaches. However, three successive extractions were required to extract all polyP. Standard addition revealed matrix effects that differed between sample types, causing polyP to be over- or underestimated by up to 50% in the samples tested here. Although previous studies judged that the presence of DNA would not complicate fluorometric quantification of polyP with DAPI, we show that RNA can cause significant interference at the wavelengths used to measure polyP. Importantly, treating samples with DNase and RNase before proteinase K digestion reduced fluorescence by up to 57%. We measured particulate polyP along a North Pacific coastal-to-open ocean transect and show that particulate polyP concentrations increased toward the open ocean. While our final method is optimized for marine particulate matter, different environmental sample types may need to be assessed for matrix effects, extraction efficiency, and nucleic acid interference. PMID:23104409

  2. Simultaneous extraction of nucleic acids and proteins from tissue specimens by ultracentrifugation: A protocol using the high-salt protein fraction for quantitative proteome analysis.

    PubMed

    Grzendowski, Michael; Riemenschneider, Markus J; Hawranke, Eva; Stefanski, Anja; Meyer, Helmut E; Reifenberger, Guido; Stühler, Kai

    2009-11-01

    Comprehensive molecular profiling of human tumor tissue specimens at the DNA, mRNA and protein level is often obstructed by a limited amount of available material. Homogenization of frozen tissue samples in guanidine isothiocyanate followed by ultracentrifugation over cesium chloride allows the simultaneous extraction of high-molecular weight DNA and RNA. Here, we present a protocol for quantitative proteome analysis using the high-salt protein fraction obtained as supernatant after ultracentrifugation for nucleic acid extraction. We applied this method to extracts from primary human brain tumors and demonstrate its successful application for protein expression profiling in these tumors using 2-D DIGE, MS and Western blotting.

  3. Application of Alternative Nucleic Acid Extraction Protocols to ProGastro SSCS Assay for Detection of Bacterial Enteric Pathogens

    PubMed Central

    Napierala, Maureen; Munson, Kimber L.; Bilbo, Dorothy; Schulte, Michael A.

    2016-01-01

    As an alternative to automated extraction, fecal specimens were processed by investigational lysis/heating (i.e., manual) and by chromatography/centrifugation (i.e., column) methods. ProGastro SSC and Shiga toxin-producing Escherichia coli (i.e., STEC) indeterminate rates for 101 specimens were 1.0% to 3.0% for automated, 11.9% for manual, and 24.8% to 37.6% for column methods. Following freeze-thaw of 247 specimens, indeterminate rates were 1.6% to 2.4% for manual and 0.8 to 5.3% for column methods. Mean processing times for manual and column methods were 30.5 and 69.2 min, respectively. Concordance of investigational methods with automated extraction was ≥98.8%. PMID:26935731

  4. Leaf tissue sampling and DNA extraction protocols.

    PubMed

    Semagn, Kassa

    2014-01-01

    Taxonomists must be familiar with a number of issues in collecting and transporting samples using freezing methods (liquid nitrogen and dry ice), desiccants (silica gel and blotter paper), and preservatives (CTAB, ethanol, and isopropanol), with each method having its own merits and limitations. For most molecular studies, a reasonably good quality and quantity of DNA is required, which can only be obtained using standard DNA extraction protocols. There are many DNA extraction protocols that vary from simple and quick ones that yield low-quality DNA but good enough for routine analyses to the laborious and time-consuming standard methods that usually produce high quality and quantities of DNA. The protocol to be chosen will depend on the quality and quantity of DNA needed, the nature of samples, and the presence of natural substances that may interfere with the extraction and subsequent analysis. The protocol described in this chapter has been tested for extracting DNA from eight species and provided very good quality and quantity of DNA for different applications, including those genotyping methods that use restriction enzymes.

  5. Extraction and anonymity protocol of medical file.

    PubMed Central

    Bouzelat, H.; Quantin, C.; Dusserre, L.

    1996-01-01

    To carry out the epidemiological study of patients suffering from a given cancer, the Department of Medical Informatics (DIM) has to link information coming from different hospitals and medical laboratories in the Burgundy region. Demands from the French department for computerized information security (Commission Nationale de l'Informatique et des Libertés: CNIL), in regard to abiding by the law of January 6, 1978, completed by the law of July 1st, 1994 on nominal data processing in the framework of medical research have to be taken into account. Notably, the CNIL advised to render anonymous patient identities before the extraction of each establishment file. This paper describes a recently implemented protocol, registered with the French department for computerized information security (Service Central de la Sécurité des Systèmes d'information : SCSSI) whose purpose is to render anonymous medical files in view of their extraction. Once rendered anonymous, these files will be exportable so as to be merged with other files and used in a framework of epidemiological studies. Therefore, this protocol uses the Standard Hash Algorithm (SHA) which allows the replacement of identities by their imprints while ensuring a minimal collision rate in order to allow a correct linkage of the different information concerning the same patient. A first evaluation of the extraction and anonymity software with regard to the purpose of an epidemiological survey is described here. In this paper, we also show how it would be possible to implement this system by means of the Internet communication network. PMID:8947681

  6. An acid-catalyzed macrolactonization protocol.

    PubMed

    Trost, Barry M; Chisholm, John D

    2002-10-17

    [reaction: see text] An efficient macrolactonization protocol devoid of any base was developed derived from the use of vinyl esters in transesterification. Subjecting a hydroxy acid and ethoxyacetylene to 2 mol % [RuCl(2)(p-cymene)](2) in toluene followed by addition of camphorsulfonic acid or inverse addition provided macrolactones in good yields.

  7. Microplastics in seafood: Benchmark protocol for their extraction and characterization.

    PubMed

    Dehaut, Alexandre; Cassone, Anne-Laure; Frère, Laura; Hermabessiere, Ludovic; Himber, Charlotte; Rinnert, Emmanuel; Rivière, Gilles; Lambert, Christophe; Soudant, Philippe; Huvet, Arnaud; Duflos, Guillaume; Paul-Pont, Ika

    2016-08-01

    Pollution of the oceans by microplastics (<5 mm) represents a major environmental problem. To date, a limited number of studies have investigated the level of contamination of marine organisms collected in situ. For extraction and characterization of microplastics in biological samples, the crucial step is the identification of solvent(s) or chemical(s) that efficiently dissolve organic matter without degrading plastic polymers for their identification in a time and cost effective way. Most published papers, as well as OSPAR recommendations for the development of a common monitoring protocol for plastic particles in fish and shellfish at the European level, use protocols containing nitric acid to digest the biological tissues, despite reports of polyamide degradation with this chemical. In the present study, six existing approaches were tested and their effects were compared on up to 15 different plastic polymers, as well as their efficiency in digesting biological matrices. Plastic integrity was evaluated through microscopic inspection, weighing, pyrolysis coupled with gas chromatography and mass spectrometry, and Raman spectrometry before and after digestion. Tissues from mussels, crabs and fish were digested before being filtered on glass fibre filters. Digestion efficiency was evaluated through microscopical inspection of the filters and determination of the relative removal of organic matter content after digestion. Five out of the six tested protocols led to significant degradation of plastic particles and/or insufficient tissue digestion. The protocol using a KOH 10% solution and incubation at 60 °C during a 24 h period led to an efficient digestion of biological tissues with no significant degradation on all tested polymers, except for cellulose acetate. This protocol appeared to be the best compromise for extraction and later identification of microplastics in biological samples and should be implemented in further monitoring studies to ensure

  8. Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols.

    PubMed

    D'Abzac, Paul; Bordas, François; Van Hullebusch, Eric; Lens, Piet N L; Guibaud, Gilles

    2010-02-01

    The characteristics of the extracellular polymeric substances (EPS) extracted with nine different extraction protocols from four different types of anaerobic granular sludge were studied. The efficiency of four physical (sonication, heating, cationic exchange resin (CER), and CER associated with sonication) and four chemical (ethylenediaminetetraacetic acid, ethanol, formaldehyde combined with heating, or NaOH) EPS extraction methods was compared to a control extraction protocols (i.e., centrifugation). The nucleic acid content and the protein/polysaccharide ratio of the EPS extracted show that the extraction does not induce abnormal cellular lysis. Chemical extraction protocols give the highest EPS extraction yields (calculated by the mass ratio between sludges and EPS dry weight (DW)). Infrared analyses as well as an extraction yield over 100% or organic carbon content over 1 g g(-1) of DW revealed, nevertheless, a carry-over of the chemical extractants into the EPS extracts. The EPS of the anaerobic granular sludges investigated are predominantly composed of humic-like substances, proteins, and polysaccharides. The EPS content in each biochemical compound varies depending on the sludge type and extraction technique used. Some extraction techniques lead to a slightly preferential extraction of some EPS compounds, e.g., CER gives a higher protein yield.

  9. Soil metaproteomics – Comparative evaluation of protein extraction protocols

    PubMed Central

    Keiblinger, Katharina M.; Wilhartitz, Inés C.; Schneider, Thomas; Roschitzki, Bernd; Schmid, Emanuel; Eberl, Leo; Riedel, Kathrin; Zechmeister-Boltenstern, Sophie

    2012-01-01

    Metaproteomics and its potential applications are very promising to study microbial activity in environmental samples and to obtain a deeper understanding of microbial interactions. However, due to the complexity of soil samples the exhaustive extraction of proteins is a major challenge. We compared soil protein extraction protocols in terms of their protein extraction efficiency for two different soil types. Four different protein extraction procedures were applied based on (a) SDS extraction without phenol, (b) NaOH and subsequent phenol extraction, (c) SDS–phenol extraction and (d) SDS–phenol extraction with prior washing steps. To assess the suitability of these methods for the functional analysis of the soil metaproteome, they were applied to a potting soil high in organic matter and a forest soil. Proteins were analyzed by two-dimensional liquid chromatography/tandem mass spectrometry (2D-LC–MS/MS) and the number of unique spectra as well as the number of assigned proteins for each of the respective protocols was compared. In both soil types, extraction with SDS–phenol (c) resulted in “high” numbers of proteins. Moreover, a spiking experiment was conducted to evaluate protein recovery. To this end sterilized forest soil was amended with proteins from pure cultures of Pectobacterium carotovorum and Aspergillus nidulans. The protein recovery in the spiking experiment was almost 50%. Our study demonstrates that a critical evaluation of the extraction protocol is crucial for the quality of the metaproteomics data, especially in highly complex samples like natural soils. PMID:23125465

  10. Soil metaproteomics - Comparative evaluation of protein extraction protocols.

    PubMed

    Keiblinger, Katharina M; Wilhartitz, Inés C; Schneider, Thomas; Roschitzki, Bernd; Schmid, Emanuel; Eberl, Leo; Riedel, Kathrin; Zechmeister-Boltenstern, Sophie

    2012-11-01

    Metaproteomics and its potential applications are very promising to study microbial activity in environmental samples and to obtain a deeper understanding of microbial interactions. However, due to the complexity of soil samples the exhaustive extraction of proteins is a major challenge. We compared soil protein extraction protocols in terms of their protein extraction efficiency for two different soil types. Four different protein extraction procedures were applied based on (a) SDS extraction without phenol, (b) NaOH and subsequent phenol extraction, (c) SDS-phenol extraction and (d) SDS-phenol extraction with prior washing steps. To assess the suitability of these methods for the functional analysis of the soil metaproteome, they were applied to a potting soil high in organic matter and a forest soil. Proteins were analyzed by two-dimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS) and the number of unique spectra as well as the number of assigned proteins for each of the respective protocols was compared. In both soil types, extraction with SDS-phenol (c) resulted in "high" numbers of proteins. Moreover, a spiking experiment was conducted to evaluate protein recovery. To this end sterilized forest soil was amended with proteins from pure cultures of Pectobacterium carotovorum and Aspergillus nidulans. The protein recovery in the spiking experiment was almost 50%. Our study demonstrates that a critical evaluation of the extraction protocol is crucial for the quality of the metaproteomics data, especially in highly complex samples like natural soils.

  11. DNA extraction from rice endosperm (including a protocol for extraction of DNA from ancient seed samples).

    PubMed

    Mutou, Chiaki; Tanaka, Katsunori; Ishikawa, Ryuji

    2014-01-01

    Deoxyribonucleic acid (DNA) extracted from endosperm can be effectively used for rapid genotyping using seed tissue, to evaluate seed quality from packaged grains and to determine the purity of milled grains. Methods outlined here are optimal procedures to isolate DNA from endosperm tissue of modern rice grains and of aged rice remains preserved between 50 and 100 years. The extracted DNA can be used to amplify regions of chloroplast genomic DNA (ctDNA), mitochondrial genomic DNA (mtDNA), and nuclear genomic DNA using standard PCR protocols. In addition, we describe an optimal procedure to process archaeological grain specimens, aged for a couple of thousand years, to isolate DNA from these ancient samples, referred to here as ancient DNA (aDNA). The aDNA can be successfully amplified by PCR using appropriate primer pairs designed specifically for aDNA amplification.

  12. Evaluation and Comparison of Four Protein Extraction Protocols for Mono- and Two-Dimensional Electrophoresis in Mytilus Galloprovincialis.

    PubMed

    Ceruso, Marina; Chirollo, Claudia; Boccia, Federica; Smaldone, Giorgio; Marrone, Raffaele; Pepe, Tiziana

    2015-06-30

    In this study, four protein extraction protocols from Mytilus galloprovincialis were evaluated with the aim to identify the most practical, efficient and reproducible method. Four extraction protocols frequently used for mussels and organic matrices were selected and compared. The methods were based on the use of: i) TRIzol reagent; ii) Lysis buffer; iii) phenylmethanesulfonyl fluoride; iv) trichloroacetic acid-acetone. Protein concentration was measured by the Bradford method. Three specimens of mussels were studied and the analysis was conducted in triplicate for each of the four protocols. Results indicated that the four methods could extract significantly different protein profiles. The highest number of protein spots resolved in 2DE gels and the best reproducibility was obtained using trichloroacetic acid-acetone protocol. Results afforded the selection of a suitable extraction protocol to be used for ecotoxicoproteomics studies from mussels and for other proteomic studies conducted by particularly complex tissues such as Mytilus galloprovincialis.

  13. Direct Cellular Lysis/Protein Extraction Protocol for Soil Metaproteomics

    SciTech Connect

    Chourey, Karuna; Jansson, Janet; Verberkmoes, Nathan C; Shah, Manesh B; Chavarria, Krystle L.; Tom, Lauren M; Brodie, Eoin L.; Hettich, Robert {Bob} L

    2010-01-01

    We present a novel direct protocol for deep proteome characterization of microorganisms in soil. The method employs thermally assisted detergent-based cellular lysis (SDS) of soil samples, followed by TCA precipitation for proteome extraction/cleanup prior to liquid chromatography-mass spectrometric characterization. This approach was developed and optimized using different soils inoculated with genome-sequenced bacteria (Gram-negative Pseudomonas putida or Gram-positive Arthrobacter chlorophenolicus). Direct soil protein extraction was compared to protein extraction from cells isolated from the soil matrix prior to lysis (indirect method). Each approach resulted in identification of greater than 500 unique proteins, with a wide range in molecular mass and functional categories. To our knowledge, this SDS-TCA approach enables the deepest proteome characterizations of microbes in soil to date, without significant biases in protein size, localization, or functional category compared to pure cultures. This protocol should provide a powerful tool for ecological studies of soil microbial communities.

  14. Direct cellular lysis/protein extraction protocol for soil metaproteomics.

    PubMed

    Chourey, Karuna; Jansson, Janet; VerBerkmoes, Nathan; Shah, Manesh; Chavarria, Krystle L; Tom, Lauren M; Brodie, Eoin L; Hettich, Robert L

    2010-12-03

    We present a novel direct protocol for deep proteome characterization of microorganisms in soil. The method employs thermally assisted detergent-based cellular lysis (SDS) of soil samples, followed by TCA precipitation for proteome extraction/cleanup prior to liquid chromatography-mass spectrometric characterization. This approach was developed and optimized using different soils inoculated with genome-sequenced bacteria (Gram-negative Pseudomonas putida or Gram-positive Arthrobacter chlorophenolicus). Direct soil protein extraction was compared to protein extraction from cells isolated from the soil matrix prior to lysis (indirect method). Each approach resulted in identification of greater than 500 unique proteins, with a wide range in molecular mass and functional categories. To our knowledge, this SDS-TCA approach enables the deepest proteome characterizations of microbes in soil to date, without significant biases in protein size, localization, or functional category compared to pure cultures. This protocol should provide a powerful tool for ecological studies of soil microbial communities.

  15. Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics.

    PubMed

    Wu, Huifeng; Ji, Chenglong; Wei, Lei; Zhao, Jianmin

    2013-11-01

    In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics.

  16. A simplified universal genomic DNA extraction protocol suitable for PCR.

    PubMed

    Wang, T Y; Wang, L; Zhang, J H; Dong, W H

    2011-03-29

    Conventional genomic DNA extraction protocols need expensive and hazardous reagents for decontamination of phenolic compounds from the extracts and are only suited for certain types of tissue. We developed a simple, time-saving and cost-efficient method for genomic DNA extraction from various types of organisms, using relatively innocuous reagents. The protocol employs a single purification step to remove contaminating compounds, using a silica column and a non-hazardous buffer, and a chaotropic-detergent lysing solution that hydrolyzes RNA and allows the selective precipitation of DNA from cell lysates. We used this system to extract genomic DNA from different tissues of various organisms, including algae (Dunaliella salina), human peripheral blood, mouse liver, Escherichia coli, and Chinese hamster ovary cells. Mean DNA yields were 20-30 μg/cm(3) from fresh tissues (comparable to yields given by commercial extraction kits), and the 260/280 nm absorbance ratio was 1.8-2.0, demonstrating a good degree of purity. The extracted DNA was successfully used in PCR, restriction enzyme digestion and for recombinant selection studies.

  17. A comparative evaluation of four DNA extraction protocols from whole blood sample.

    PubMed

    Ghaheri, M; Kahrizi, D; Yari, K; Babaie, A; Suthar, R S; Kazemi, E

    2016-03-31

    All organisms have Deoxyribonucleic acid (DNA) within their cells. DNA is a complex molecule that contains all of the information necessary to build and maintain an organism. DNA extraction is one of the most basic and essential techniques in the study of DNA that allow huge advances in molecular biology, biotechnology and bioinformatics laboratories. Whole blood samples are one of the main sources used to obtain DNA and there are many different protocols available in this issue. In current research, compared four DNA extraction protocols from blood samples; include modified phenol-chloroform protocol, two salting-out and enzyme free method and from commercial kit. The extracted DNAs by these protocols were analyzed according to their time demands, quality and quantity, toxicity and functionality in PCR method. Also the quality and quantity of the extracted DNA were surveyed by gel electrophoresis and Nanodrop spectrophotometry methods. It was observed that there are not significantly differences between these methods about DNA Purity (A260/A280), but the DNA yield (ng DNA/μl) of phenol/chloroform method was higher than other methods. In addition, phenol/chloroform was the most toxic method and it takes more time than other methods. Roche diagnostics GmbH kit was the most expensive among the four methods but the least extraction time was required and it was the safest method.

  18. Comparative analysis of protocols for DNA extraction from soybean caterpillars.

    PubMed

    Palma, J; Valmorbida, I; da Costa, I F D; Guedes, J V C

    2016-04-07

    Genomic DNA extraction is crucial for molecular research, including diagnostic and genome characterization of different organisms. The aim of this study was to comparatively analyze protocols of DNA extraction based on cell lysis by sarcosyl, cetyltrimethylammonium bromide, and sodium dodecyl sulfate, and to determine the most efficient method applicable to soybean caterpillars. DNA was extracted from specimens of Chrysodeixis includens and Spodoptera eridania using the aforementioned three methods. DNA quantification was performed using spectrophotometry and high molecular weight DNA ladders. The purity of the extracted DNA was determined by calculating the A260/A280 ratio. Cost and time for each DNA extraction method were estimated and analyzed statistically. The amount of DNA extracted by these three methods was sufficient for PCR amplification. The sarcosyl method yielded DNA of higher purity, because it generated a clearer pellet without viscosity, and yielded high quality amplification products of the COI gene I. The sarcosyl method showed lower cost per extraction and did not differ from the other methods with respect to preparation times. Cell lysis by sarcosyl represents the best method for DNA extraction in terms of yield, quality, and cost effectiveness.

  19. An innovative protocol for schwann cells extracellular matrix proteins extraction.

    PubMed

    Parisi, L; Zomer Volpato, F; Cagol, N; Siciliano, M; Migliaresi, C; Motta, A; Sala, R

    2016-12-01

    The evidence that extracellular matrix (ECM) components could represent new targets for drugs designed to approach degenerative disease, requires their analysis. Before the analysis, proteins should be extracted from ECM and solubilized. Currently, few protocols for ECM proteins extraction and solubilization are available in literature, and most of them are based mainly on the use of proteolytic enzymes, such as trypsin, which often lead to proteins damage. Moreover, no methods have been so far proposed to solubilize Schwann Cell ECM, which may represent an important target for the therapy of neurodegenerative disorders. In our study, we propose to solubilize SC ECM through the use of surfactants and urea. We compared our method of solubilization, with one of that proposed in literature for a general ECM, mainly based on the use of enzymes. We want to highlight the benefit of solubilizing SC ECM, avoiding the use of proteolytic enzymes. To compare the amount of proteins extracted with both methods, MicroBCA assay was used, while the quality of the proteins extracted was observed through the SDS-PAGE. The results obtained confirm a better solubilization of SC ECM proteins with the proposed protocol, both quantitatively and qualitatively, showing a higher concentration of proteins extracted and a better enrichment of protein fractions, if compared to the enzyme-based protocol. Our results show that SC ECM could be efficiently solubilized through the use of surfactant and urea, avoiding the use of enzyme-base methods. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3175-3180, 2016.

  20. Extractive fermentation of acetic acid

    SciTech Connect

    Busche, R.M.

    1991-12-31

    In this technoeconomic evaluation of the manufacture of acetic acid by fermentation, the use of the bacterium: Acetobacter suboxydans from the old vinegar process was compared with expected performance of the newer Clostridium thermoaceticum bacterium. Both systems were projected to operate as immobilized cells in a continuous, fluidized bed bioreactor, using solvent extraction to recover the product. Acetobacter metabolizes ethanol aerobically to produce acid at 100 g/L in a low pH medium. This ensures that the product is in the form of a concentrated extractable free acid, rather than as an unextractable salt. Unfortunately, yields from glucose by way of the ethanol fermentation are poor, but near the biological limits of the organisms involved. Conversely, C. thermoaceticum is a thermophilic anaerobe that operates at high fermentation rates on glucose at neutral pH to produce acetate salts directly in substantially quantitative yields. However, it is severely inhibited by product, which restricts concentration to a dilute 20 g/L. An improved Acetobacter system operating with recycled cells at 50 g/L appears capable of producing acid at $0.38/lb, as compared with a $0.29/lb price for synthetic acid. However, this system has only a limited margin for process improvement. The present Clostridium system cannot compete, since the required selling price would be $0.42/lb. However, if the organism could be adapted to tolerate higher product concentrations at acid pH, selling price could be reduced to $0.22/lb, or about 80% of the price of synthetic acid.

  1. Subcritical Water Extraction of Amino Acids from Atacama Desert Soils

    NASA Technical Reports Server (NTRS)

    Amashukeli, Xenia; Pelletier, Christine C.; Kirby, James P.; Grunthaner, Frank J.

    2007-01-01

    Amino acids are considered organic molecular indicators in the search for extant and extinct life in the Solar System. Extraction of these molecules from a particulate solid matrix, such as Martian regolith, will be critical to their in situ detection and analysis. The goals of this study were to optimize a laboratory amino acid extraction protocol by quantitatively measuring the yields of extracted amino acids as a function of liquid water temperature and sample extraction time and to compare the results to the standard HCl vapor- phase hydrolysis yields for the same soil samples. Soil samples from the Yungay region of the Atacama Desert ( Martian regolith analog) were collected during a field study in the summer of 2005. The amino acids ( alanine, aspartic acid, glutamic acid, glycine, serine, and valine) chosen for analysis were present in the samples at concentrations of 1 - 70 parts- per- billion. Subcritical water extraction efficiency was examined over the temperature range of 30 - 325 degrees C, at pressures of 17.2 or 20.0 MPa, and for water- sample contact equilibration times of 0 - 30 min. None of the amino acids were extracted in detectable amounts at 30 degrees C ( at 17.2 MPa), suggesting that amino acids are too strongly bound by the soil matrix to be extracted at such a low temperature. Between 150 degrees C and 250 degrees C ( at 17.2 MPa), the extraction efficiencies of glycine, alanine, and valine were observed to increase with increasing water temperature, consistent with higher solubility at higher temperatures, perhaps due to the decreasing dielectric constant of water. Amino acids were not detected in extracts collected at 325 degrees C ( at 20.0 MPa), probably due to amino acid decomposition at this temperature. The optimal subcritical water extraction conditions for these amino acids from Atacama Desert soils were achieved at 200 degrees C, 17.2 MPa, and a water- sample contact equilibration time of 10 min.

  2. Effective protein extraction protocol for proteomics studies of Jerusalem artichoke leaves.

    PubMed

    Zhang, Meide; Shen, Shihua

    2013-07-01

    Protein extraction is a crucial step for proteomics studies. To establish an effective protein extraction protocol suitable for two-dimensional electrophoresis (2DE) analysis in Jerusalem artichoke (Helianthus tuberosus L.), three different protein extraction methods-trichloroacetic acid/acetone, Mg/NP-40, and phenol/ammonium acetate-were evaluated using Jerusalem artichoke leaves as source materials. Of the three methods, trichloroacetic acid/acetone yielded the best protein separation pattern and highest number of protein spots in 2DE analysis. Proteins highly abundant in leaves, such as Rubisco, are typically problematic during leaf 2DE analysis, however, and this disadvantage was evident using trichloroacetic acid/acetone. To reduce the influence of abundant proteins on the detection of low-abundance proteins, we optimized the trichloroacetic acid/acetone method by incorporating a PEG fractionation approach. After optimization, 363 additional (36.2%) protein spots were detected on the 2DE gel. Our results suggest that trichloroacetic acid/acetone method is a better protein extraction technique than Mg/NP-40 and phenol/ammonium acetate in Jerusalem artichoke leaf 2DE analysis, and that trichloroacetic acid/acetone method combined with PEG fractionation procedure is the most effective approach for leaf 2DE analysis of Jerusalem artichoke.

  3. Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods

    PubMed Central

    Tambellini, Nicolas P.; Zaremberg, Vanina; Turner, Raymond J.; Weljie, Aalim M.

    2013-01-01

    Metabolomic and lipidomic approaches aim to measure metabolites or lipids in the cell. Metabolite extraction is a key step in obtaining useful and reliable data for successful metabolite studies. Significant efforts have been made to identify the optimal extraction protocol for various platforms and biological systems, for both polar and non-polar metabolites. Here we report an approach utilizing chemoinformatics for systematic comparison of protocols to extract both from a single sample of the model yeast organism Saccharomyces cerevisiae. Three chloroform/methanol/water partitioning based extraction protocols found in literature were evaluated for their effectiveness at reproducibly extracting both polar and non-polar metabolites. Fatty acid methyl esters and methoxyamine/trimethylsilyl derivatized aqueous compounds were analyzed by gas chromatography mass spectrometry to evaluate non-polar or polar metabolite analysis. The comparative breadth and amount of recovered metabolites was evaluated using multivariate projection methods. This approach identified an optimal protocol consisting of 64 identified polar metabolites from 105 ion hits and 12 fatty acids recovered, and will potentially attenuate the error and variation associated with combining metabolite profiles from different samples for untargeted analysis with both polar and non-polar analytes. It also confirmed the value of using multivariate projection methods to compare established extraction protocols. PMID:24958140

  4. Evaluation of extraction protocols for simultaneous polar and non-polar yeast metabolite analysis using multivariate projection methods.

    PubMed

    Tambellini, Nicolas P; Zaremberg, Vanina; Turner, Raymond J; Weljie, Aalim M

    2013-07-23

    Metabolomic and lipidomic approaches aim to measure metabolites or lipids in the cell. Metabolite extraction is a key step in obtaining useful and reliable data for successful metabolite studies. Significant efforts have been made to identify the optimal extraction protocol for various platforms and biological systems, for both polar and non-polar metabolites. Here we report an approach utilizing chemoinformatics for systematic comparison of protocols to extract both from a single sample of the model yeast organism Saccharomyces cerevisiae. Three chloroform/methanol/water partitioning based extraction protocols found in literature were evaluated for their effectiveness at reproducibly extracting both polar and non-polar metabolites. Fatty acid methyl esters and methoxyamine/trimethylsilyl derivatized aqueous compounds were analyzed by gas chromatography mass spectrometry to evaluate non-polar or polar metabolite analysis. The comparative breadth and amount of recovered metabolites was evaluated using multivariate projection methods. This approach identified an optimal protocol consisting of 64 identified polar metabolites from 105 ion hits and 12 fatty acids recovered, and will potentially attenuate the error and variation associated with combining metabolite profiles from different samples for untargeted analysis with both polar and non-polar analytes. It also confirmed the value of using multivariate projection methods to compare established extraction protocols.

  5. A simple Chelex protocol for DNA extraction from Anopheles spp.

    PubMed

    Musapa, Mulenga; Kumwenda, Taida; Mkulama, Mtawa; Chishimba, Sandra; Norris, Douglas E; Thuma, Philip E; Mharakurwa, Sungano

    2013-01-09

    Endemic countries are increasingly adopting molecular tools for efficient typing, identification and surveillance against malaria parasites and vector mosquitoes, as an integral part of their control programs. For sustainable establishment of these accurate approaches in operations research to strengthen malaria control and elimination efforts, simple and affordable methods, with parsimonious reagent and equipment requirements are essential. Here we present a simple Chelex-based technique for extracting malaria parasite and vector DNA from field collected mosquito specimens. We morphologically identified 72 Anopheles gambiae sl. from 156 mosquitoes captured by pyrethrum spray catches in sleeping rooms of households within a 2,000 km(2) vicinity of the Malaria Institute at Macha. After dissection to separate the head and thorax from the abdomen for all 72 Anopheles gambiae sl. mosquitoes, the two sections were individually placed in 1.5 ml microcentrifuge tubes and submerged in 20 μl of deionized water. Using a sterile pipette tip, each mosquito section was separately homogenized to a uniform suspension in the deionized water. Of the ensuing homogenate from each mosquito section, 10 μl was retained while the other 10 μl was transferred to a separate autoclaved 1.5 ml tube. The separate aliquots were subjected to DNA extraction by either the simplified Chelex or the standard salting out extraction protocol(9,10). The salting out protocol is so-called and widely used because it employs high salt concentrations in lieu of hazardous organic solvents (such as phenol and chloroform) for the protein precipitation step during DNA extraction(9). Extracts were used as templates for PCR amplification using primers targeting arthropod mitochondrial nicotinamide adenine dinucleotide dehydrogenase (NADH) subunit 4 gene (ND4) to check DNA quality, a PCR for identification of Anopheles gambiae sibling species(10) and a nested PCR for typing of Plasmodium falciparum infection

  6. DNA extraction protocols from dormant buds of twelve woody plant genera

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Standard plant DNA extraction protocols call for samples of newly expanding leaves and shoots yet analysis is sometimes needed when plants are dormant. We evaluated three DNA extraction protocols using dormant buds from 40 species and four hybrids of 12 genera. Two protocols were from ready-to-use ...

  7. Development of a standardized sequential extraction protocol for simultaneous extraction of multiple actinide elements

    DOE PAGES

    Faye, Sherry A.; Richards, Jason M.; Gallardo, Athena M.; ...

    2017-02-07

    Sequential extraction is a useful technique for assessing the potential to leach actinides from soils; however, current literature lacks uniformity in experimental details, making direct comparison of results impossible. This work continued development toward a standardized five-step sequential extraction protocol by analyzing extraction behaviors of 232Th, 238U, 239,240Pu and 241Am from lake and ocean sediment reference materials. Results produced a standardized procedure after creating more defined reaction conditions to improve method repeatability. A NaOH fusion procedure is recommended following sequential leaching for the complete dissolution of insoluble species.

  8. Solvent extraction of metals with hydroxamic acids.

    PubMed

    Vernon, F; Khorassani, J H

    1978-07-01

    Solvent extraction with hydroxamic acids has been investigated. with comparison of aliphatic and aromatic reagents for the extraction of iron, copper, cobalt and nickel. Caprylohydroxamic acid has been evaluated for use in extraction systems for titanium, vanadium, chromium, molybdenum and uranium, both in terms of acidity of aqueous phase and oxidation state of the metal. It has been established that caprylohydroxamic acid in 1-hexanol is a suitable extractant for the removal of titanium(IV), vanadium(V), chromium(VI), molybdenum(VI) and uranium(VI) from 6M hydrochloric acid.

  9. A universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis.

    PubMed

    Wang, Wei; Vignani, Rita; Scali, Monica; Cresti, Mauro

    2006-07-01

    A simple and universally applicable protocol for extracting high-quality proteins from recalcitrant plant tissues is described. We have used the protocol with no modification, for a wide range of leaves and fruits. In all cases, this protocol allows to obtain good electrophoretic separation of proteins. As the protocol is rapid, universal, and compatible with silver staining, it could be used for routine protein extraction from recalcitrant plant tissues for proteomic analysis.

  10. Microwave-Assisted γ-Valerolactone Production for Biomass Lignin Extraction: A Cascade Protocol.

    PubMed

    Tabasso, Silvia; Grillo, Giorgio; Carnaroglio, Diego; Calcio Gaudino, Emanuela; Cravotto, Giancarlo

    2016-03-26

    The general need to slow the depletion of fossil resources and reduce carbon footprints has led to tremendous effort being invested in creating "greener" industrial processes and developing alternative means to produce fuels and synthesize platform chemicals. This work aims to design a microwave-assisted cascade process for a full biomass valorisation cycle. GVL (γ-valerolactone), a renewable green solvent, has been used in aqueous acidic solution to achieve complete biomass lignin extraction. After lignin precipitation, the levulinic acid (LA)-rich organic fraction was hydrogenated, which regenerated the starting solvent for further biomass delignification. This process does not requires a purification step because GVL plays the dual role of solvent and product, while the reagent (LA) is a product of biomass delignification. In summary, this bio-refinery approach to lignin extraction is a cascade protocol in which the solvent loss is integrated into the conversion cycle, leading to simplified methods for biomass valorisation.

  11. A Robust Protocol for Protein Extraction and Digestion.

    PubMed

    Atallah, Michelle; Flory, Mark R; Mallick, Parag

    2017-01-01

    Proteins play a key role in all aspects of cellular homeostasis. Proteomics, the large-scale study of proteins, provides in-depth data on protein properties, including abundances and post-translational modification states, and as such provides a rich avenue for the investigation of biological and disease processes. While proteomic tools such as mass spectrometry have enabled exquisitely sensitive sample analysis, sample preparation remains a critical unstandardized variable that can have a significant impact on downstream data readouts. Consistency in sample preparation and handling is therefore paramount in the collection and analysis of proteomic data.Here we describe methods for performing protein extraction from cell culture or tissues, digesting the isolated protein into peptides via in-solution enzymatic digest, and peptide cleanup with final preparations for analysis via liquid chromatography-mass spectrometry. These protocols have been optimized and standardized for maximum consistency and maintenance of sample integrity.

  12. Evaluation of Olive Fruit Lipoxygenase Extraction Protocols on 9- and 13-Z,E-HPODE Formation.

    PubMed

    Soldo, Barbara; Šprung, Matilda; Mušac, Gloria; Pavela-Vrančić, Maja; Ljubenkov, Ivica

    2016-04-20

    In plant tissues, enzymes implicated in the lipoxygenase (LOX) pathway are responsible for the hydroperoxydation of polyunsaturated fatty acids, ultimately leading to the production of small chemical species involved in several physiological processes. During industrial olive oil production, these enzymes are activated upon crushing and grinding of olive fruit tissue, subsequently leading to the synthesis of volatile compounds responsible for the positive aroma and flavor of the oil. An investigation of LOX activity during olive fruit ripening and malaxation could assist in the production of oils with favorable aroma and taste. Therefore, a reliable method for olive LOX purification is crucial. Here we report a critical review of six LOX extraction protocols, two of which have shown minimum enzyme activity, possibly leading to misconceptions in the interpretation of experimental data. Future research concerning olive LOX should employ extraction methods that preserve enzyme activity.

  13. A simple protocol for extracting hemocytes from wild caterpillars.

    PubMed

    Stoepler, Teresa M; Castillo, Julio C; Lill, John T; Eleftherianos, Ioannis

    2012-11-15

    Insect hemocytes (equivalent to mammalian white blood cells) play an important role in several physiological processes throughout an insect's life cycle. In larval stages of insects belonging to the orders of Lepidoptera (moths and butterflies) and Diptera (true flies), hemocytes are formed from the lymph gland (a specialized hematopoietic organ) or embryonic cells and can be carried through to the adult stage. Embryonic hemocytes are involved in cell migration during development and chemotaxis regulation during inflammation. They also take part in cell apoptosis and are essential for embryogenesis. Hemocytes mediate the cellular arm of the insect innate immune response that includes several functions, such as cell spreading, cell aggregation, formation of nodules, phagocytosis and encapsulation of foreign invaders. They are also responsible for orchestrating specific insect humoral defenses during infection, such as the production of antimicrobial peptides and other effector molecules. Hemocyte morphology and function have mainly been studied in genetic or physiological insect models, including the fruit fly, Drosophila melanogaster, the mosquitoes Aedes aegypti and Anopheles gambiae and the tobacco hornworm, Manduca sexta. However, little information currently exists about the diversity, classification, morphology and function of hemocytes in non-model insect species, especially those collected from the wild. Here we describe a simple and efficient protocol for extracting hemocytes from wild caterpillars. We use penultimate instar Lithacodes fasciola (yellow-shouldered slug moth) (Figure 1) and Euclea delphinii (spiny oak slug) caterpillars (Lepidoptera: Limacodidae) and show that sufficient volumes of hemolymph (insect blood) can be isolated and hemocyte numbers counted from individual larvae. This method can be used to efficiently study hemocyte types in these species as well as in other related lepidopteran caterpillars harvested from the field, or it can be

  14. Extraction chemistry of fermentation product carboxylic acid

    SciTech Connect

    Kertes, A.S.; King, C.J.

    1986-02-01

    Within the framework of a program aiming to improve the existing extractive recovery technology of fermentation products, the state of the art is critically reviewed. The acids under consideration are propionic, lactic, pyruvic, succinic, fumaric, maleic, malic, itaconic, tartaric, citric, and isocitric, all obtained by the aerobic fermentation of glucose via the glycolytic pathway and glyoxylate bypass. With no exception, it is the undissociated monomeric acid that is extracted into carbon-bonded and phosphorus-bonded oxygen donor extractants. In the organic phase, the acids are usually dimerized. The extractive transfer process obeys the Nernst law, and the measured partition coefficients range from about 0.003 for aliphatic hydrocarbons to about 2 to 3 for aliphatic alcohols and ketones to about 10 or more for organophosphates. Equally high distribution ratios are measured when long-chain tertiary amines are employed as extractants, forming bulky salts preferentially soluble in the organic phase. 123 references.

  15. Extraction chemistry of fermentation product carboxylic acids

    SciTech Connect

    Kertes, A.S.; King, C.J.

    1986-02-01

    Within the framework of a program aiming to improve the existing extractive recovery technology of fermentation products, the state of the art is critically reviewed. The acids under consideration are propionic, lactic, pyruvic, succinic, fumaric, maleic, malic, itaconic, tartaric, citric, and isocitric, all obtained by the aerobic fermentation of glucose via the glycolytic pathways and glyoxylate bypass. With no exception, it is the undissociated monomeric acid that is extracted into carbon-bonded and phosphorus-bonded oxygen donor extractants. In the organic phase, the acids are usually dimerized. The extractive transfer process obeys the Nernst law, and the measured partition coefficients range from about 0.003 for aliphatic hydrocarbons to about 2 to 3 for aliphatic alcohols and ketones to about 10 or more for organophosphates. Equally high distribution ratios are measured when long-chain tertiary amines are employed as extractants, forming bulky salts preferentially soluble in the organic phase.

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

    PubMed

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

    2014-02-01

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

  17. A high yield multi-method extraction protocol for protein quantification in activated sludge.

    PubMed

    Monique, Ras; Elisabeth, Girbal-Neuhauser; Etienne, Paul; Dominique, Lefebvre

    2008-11-01

    A multi-method extraction protocol based on mechanical, ionic and hydrophobic methods was investigated on two types of activated sludge samples. Extraction methods were chosen with regards to optimal protein yield without cell disruption. Sonication, EDTA and Tween extraction methods were selected and combined. The total amount of protein released by the multi-method protocol sums up to 191 and 264 mg equiv. BSA/g VSS for the two different sludge samples. Protocol repetition on the same sample showed that protein yield after each successive protocol fitted an exponential curve model. The total amount of extractable proteins was evaluated by model predictions, 423 and 516 mg equiv. BSA/g VSS for the two sludge samples. The multi-method extraction protocol appears relevant for harvesting a representative quantity of proteins from the original sample (45-49%), moreover the multi-method criterion of the protocol also offers a heterogeneous pool of proteins. Thus, further qualitative studies may not be biased by the extraction protocol.

  18. A DNA extraction protocol for improved DNA yield from individual mosquitoes

    PubMed Central

    Nieman, Catelyn C.; Yamasaki, Youki; Collier, Travis C.; Lee, Yoosook

    2015-01-01

    Typical DNA extraction protocols from commercially available kits provide an adequate amount of DNA from a single individual mosquito sufficient for PCR-based assays. However, next-generation sequencing applications and high-throughput SNP genotyping assays exposed the limitation of DNA quantity one usually gets from a single individual mosquito. Whole genome amplification could alleviate the issue but it also creates bias in genome representation. While trying to find alternative DNA extraction protocols for improved DNA yield, we found that a combination of the tissue lysis protocol from Life Technologies and the DNA extraction protocol from Qiagen yielded a higher DNA amount than the protocol using the Qiagen or Life Technologies kit only. We have not rigorously tested all the possible combinations of extraction protocols; we also only tested this on mosquito samples. Therefore, our finding should be noted as a suggestion for improving people’s own DNA extraction protocols and not as an advertisement of a commercially available product. PMID:26937269

  19. PROCESS FOR PRODUCING ALKYL ORTHOPHOSPHORIC ACID EXTRACTANTS

    DOEpatents

    Grinstead, R.R.

    1962-01-23

    A process is given for producing superior alkyl orthophosphoric acid extractants for use in solvent extraction methods to recover and purify various metals such as uranium and vanadium. The process comprises slurrying P/sub 2/O/ sub 5/ in a solvent diluent such as kerosene, benzene, isopropyl ether, and the like. An alipbatic alcohol having from nine to seventeen carbon atoms, and w- hcrein ihc OH group is situated inward of the terminal carbon atoms, is added to the slurry while the reaction temperature is mainiained below 60 deg C. The alcohol is added in the mole ratio of about 2 to l, alcohol to P/sub 2/O/sub 5/. A pyrophosphate reaotion product is formed in the slurry-alcohol mixture. Subsequently, the pyrophosphate reaction product is hydrolyzed with dilute mineral acid to produce the desired alkyl orthophosphoric aeid extractant. The extraetant may then be separated and utilized in metal-recovery, solvent- extraction processes. (AEC)

  20. Alkaline earth cation extraction from acid solution

    DOEpatents

    Dietz, Mark; Horwitz, E. Philip

    2003-01-01

    An extractant medium for extracting alkaline earth cations from an aqueous acidic sample solution is described as are a method and apparatus for using the same. The separation medium is free of diluent, free-flowing and particulate, and comprises a Crown ether that is a 4,4'(5')[C.sub.4 -C.sub.8 -alkylcyclohexano]18-Crown-6 dispersed on an inert substrate material.

  1. Bridging the gap between comprehensive extraction protocols in plant metabolomics studies and method validation.

    PubMed

    Bijttebier, Sebastiaan; Van der Auwera, Anastasia; Foubert, Kenn; Voorspoels, Stefan; Pieters, Luc; Apers, Sandra

    2016-09-07

    It is vital to pay much attention to the design of extraction methods developed for plant metabolomics, as any non-extracted or converted metabolites will greatly affect the overall quality of the metabolomics study. Method validation is however often omitted in plant metabolome studies, as the well-established methodologies for classical targeted analyses such as recovery optimization cannot be strictly applied. The aim of the present study is to thoroughly evaluate state-of-the-art comprehensive extraction protocols for plant metabolomics with liquid chromatography-photodiode array-accurate mass mass spectrometry (LC-PDA-amMS) by bridging the gap with method validation. Validation of an extraction protocol in untargeted plant metabolomics should ideally be accomplished by validating the protocol for all possible outcomes, i.e. for all secondary metabolites potentially present in the plant. In an effort to approach this ideal validation scenario, two plant matrices were selected based on their wide versatility of phytochemicals: meadowsweet (Filipendula ulmaria) for its polyphenols content, and spicy paprika powder (from the genus Capsicum) for its apolar phytochemicals content (carotenoids, phytosterols, capsaicinoids). These matrices were extracted with comprehensive extraction protocols adapted from literature and analysed with a generic LC-PDA-amMS characterization platform that was previously validated for broad range phytochemical analysis. The performance of the comprehensive sample preparation protocols was assessed based on extraction efficiency, repeatability and intermediate precision and on ionization suppression/enhancement evaluation. The manuscript elaborates on the finding that none of the extraction methods allowed to exhaustively extract the metabolites. Furthermore, it is shown that depending on the extraction conditions enzymatic degradation mechanisms can occur. Investigation of the fractions obtained with the different extraction methods

  2. Comparison of three detergent-free protein extraction protocols for white adipose tissue.

    PubMed

    Sajic, Tatjana; Hopfgartner, Gérard; Szanto, Ildiko; Varesio, Emmanuel

    2011-08-15

    A comparative study of three detergent-free protein extraction protocols--a differential centrifugal fractionation, a delipidation protocol based on the Bligh and Dyer method, and the trifluoroethanol addition as cosolvent to an aqueous buffer--was performed on white adipose tissue. The performance of the protocols directly compatible with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was evaluated based on the total protein extraction yield and the protein recovery from different functional and cellular compartments. The most suitable method for the extraction of white adipose tissue proteins from a wide range of cellular and structural compartments was the delipidation protocol based on the Bligh and Dyer method.

  3. Optimizing Standard Sequential Extraction Protocol With Lake And Ocean Sediments

    EPA Science Inventory

    The environmental mobility/availability behavior of radionuclides in soils and sediments depends on their speciation. Experiments have been carried out to develop a simple but robust radionuclide sequential extraction method for identification of radionuclide partitioning in sed...

  4. Comparison of protocols for DNA extraction from long-term preserved formalin fixed tissues.

    PubMed

    Paireder, Stefan; Werner, Bettina; Bailer, Josef; Werther, Wolfgang; Schmid, Erich; Patzak, Beatrix; Cichna-Markl, Margit

    2013-08-15

    The current study compared the applicability of protocols to extract DNA from formalin fixed heart tissues that have been preserved for more than 50 years. Ten methods were tested: a cetyltrimethylammonium bromide (CTAB) standard protocol, seven variants of this standard protocol, and two commercial kits. In the case of younger specimens (fixed in 1951, 1934, or 1914), extracts with DNA concentrations ≥ 10.0 ng/μl were obtained with the standard CTAB protocol, two variants of the standard protocol including prolonged tissue digestion (72 h instead of 1-2h), and a commercial kit particularly recommended for DNA extraction from formalin fixed paraffin embedded tissues (FFPE Kit). With the FFPE Kit, DNA could also be extracted from older tissues (fixed in 1893, 1850/1851, or before 1820). In general, the purity of the DNA extracts, assessed from the ratio of the absorbance at 260 and 280 nm, was not very high. In spite of their rather low purity, the DNA extracts could, however, be used to amplify a 122-bp sequence and, in most cases, also a 171-bp sequence of the gene coding for human albumin by the polymerase chain reaction (PCR).

  5. Tracing tree nut allergens in chocolate: A comparison of DNA extraction protocols.

    PubMed

    Costa, Joana; Melo, Vítor S; Santos, Cristina G; Oliveira, M Beatriz P P; Mafra, Isabel

    2015-11-15

    The present work aimed at comparing different DNA extraction methods, from chocolate matrices, for the effective application in molecular techniques to detect tree nut allergens. For this study, DNA from almond or hazelnut model chocolates was extracted using seven selected protocols: the in-house methods of CTAB-PVP (cetyltrimethylammonium bromide-polyvinylpyrrolidone), Wizard with and without RNase, Wizard-PVP with and without RNase, and the Wizard Magnetic and Nucleospin kits. The extracts were assessed for their suitability for amplification by qualitative PCR and real-time PCR. From the evaluated protocols, Nucleospin presented the best results for almond and hazelnut amplification, achieving a limit of detection of 0.005% (w/w) with high PCR efficiency, linearity and range of amplification. These results highlight the importance of the DNA extraction protocol in the case of food allergens from complex matrices, such as chocolate, in which sensitivity is a key parameter.

  6. Sodium sulphite inhibition of potato and cherry polyphenolics in nucleic acid extraction for virus detection by RT-PCR.

    PubMed

    Singh, R P; Nie, X; Singh, M; Coffin, R; Duplessis, P

    2002-01-01

    Phenolic compounds from plant tissues inhibit reverse transcription-polymerase chain reaction (RT-PCR). Multiple-step protocols using several additives to inhibit polyphenolic compounds during nucleic acid extraction are common, but time consuming and laborious. The current research highlights that the inclusion of 0.65 to 0.70% of sodium sulphite in the extraction buffer minimizes the pigmentation of nucleic acid extracts and improves the RT-PCR detection of Potato virus Y (PVY) and Potato leafroll virus (PLRV) in potato (Solanum tuberosum) tubers and Prune dwarf virus (PDV) and Prunus necrotic ringspot virus (PNRSV) in leaves and bark in the sweet cherry (Prunus avium) tree. Substituting sodium sulphite in the nucleic acid extraction buffer eliminated the use of proteinase K during extraction. Reagents phosphate buffered saline (PBS)-Tween 20 and polyvinylpyrrolidone (PVP) were also no longer required during RT or PCR phase. The resultant nucleic acid extracts were suitable for both duplex and multiplex RT-PCR. This simple and less expensive nucleic acid extraction protocol has proved very effective for potato cv. Russet Norkotah, which contains a high amount of polyphenolics. Comparing commercially available RNA extraction kits (Catrimox and RNeasy), the sodium sulphite based extraction protocol yielded two to three times higher amounts of RNA, while maintaining comparable virus detection by RT-PCR. The sodium sulphite based extraction protocol was equally effective in potato tubers, and in leaves and bark from the cherry tree.

  7. A high-throughput, high-quality plant genomic DNA extraction protocol.

    PubMed

    Li, H; Li, J; Cong, X H; Duan, Y B; Li, L; Wei, P C; Lu, X Z; Yang, J B

    2013-10-15

    The isolation of high-quality genomic DNA (gDNA) is a crucial technique in plant molecular biology. The quality of gDNA determines the reliability of real-time polymerase chain reaction (PCR) analysis. In this paper, we reported a high-quality gDNA extraction protocol optimized for real-time PCR in a variety of plant species. Performed in a 96-well block, our protocol provides high throughput. Without the need for phenol-chloroform and liquid nitrogen or dry ice, our protocol is safer and more cost-efficient than traditional DNA extraction methods. The method takes 10 mg leaf tissue to yield 5-10 µg high-quality gDNA. Spectral measurement and electrophoresis were used to demonstrate gDNA purity. The extracted DNA was qualified in a restriction enzyme digestion assay and conventional PCR. The real-time PCR amplification was sufficiently sensitive to detect gDNA at very low concentrations (3 pg/µL). The standard curve of gDNA dilutions from our phenol-chloroform-free protocol showed better linearity (R(2) = 0.9967) than the phenol-chloroform protocol (R(2) = 0.9876). The results indicate that the gDNA was of high quality and fit for real-time PCR. This safe, high-throughput plant gDNA extraction protocol could be used to isolate high-quality gDNA for real-time PCR and other downstream molecular applications.

  8. Comparative assessment of various lipid extraction protocols and optimization of transesterification process for microalgal biodiesel production.

    PubMed

    Mandal, Shovon; Patnaik, Reeza; Singh, Amit Kumar; Mallick, Nirupama

    2013-01-01

    Biodiesel, using microalgae as feedstocks, is being explored as the most potent form of alternative diesel fuel for sustainable economic development. A comparative assessment of various protocols for microalgal lipid extraction was carried out using five green algae, six blue-green algae and two diatom species treated with different single and binary solvents both at room temperature and using a soxhlet. Lipid recovery was maximum with chloroform-methanol in the soxhlet extractor. Pretreatments ofbiomass, such as sonication, homogenization, bead-beating, lyophilization, autoclaving, microwave treatment and osmotic shock did not register any significant rise in lipid recovery. As lipid recovery using chloroform-methanol at room temperature demonstrated a marginally lower value than that obtained under the soxhlet extractor, on economical point of view, the former is recommended for microalgal total lipid extraction. Transesterification process enhances the quality of biodiesel. Experiments were designed to determine the effects of catalyst type and quantity, methanol to oil ratio, reaction temperature and time on the transesterification process using response surface methodology. Fatty acid methyl ester yield reached up to 91% with methanol:HCl:oil molar ratio of 82:4:1 at 65 degrees C for 6.4h reaction time. The biodiesel yield relative to the weight of the oil was found to be 69%.

  9. Protocol: A high-throughput DNA extraction system suitable for conifers

    PubMed Central

    Bashalkhanov, Stanislav; Rajora, Om P

    2008-01-01

    Background High throughput DNA isolation from plants is a major bottleneck for most studies requiring large sample sizes. A variety of protocols have been developed for DNA isolation from plants. However, many species, including conifers, have high contents of secondary metabolites that interfere with the extraction process or the subsequent analysis steps. Here, we describe a procedure for high-throughput DNA isolation from conifers. Results We have developed a high-throughput DNA extraction protocol for conifers using an automated liquid handler and modifying the Qiagen MagAttract Plant Kit protocol. The modifications involve change to the buffer system and improving the protocol so that it almost doubles the number of samples processed per kit, which significantly reduces the overall costs. We describe two versions of the protocol: one for medium-throughput (MTP) and another for high-throughput (HTP) DNA isolation. The HTP version works from start to end in the industry-standard 96-well format, while the MTP version provides higher DNA yields per sample processed. We have successfully used the protocol for DNA extraction and genotyping of thousands of individuals of several spruce and a pine species. Conclusion A high-throughput system for DNA extraction from conifer needles and seeds has been developed and validated. The quality of the isolated DNA was comparable with that obtained from two commonly used methods: the silica-spin column and the classic CTAB protocol. Our protocol provides a fully automatable and cost effective solution for processing large numbers of conifer samples. PMID:18673554

  10. Structural feature extraction protocol for classifying reversible membrane binding protein domains.

    PubMed

    Källberg, Morten; Lu, Hui

    2009-01-01

    Machine learning based classification protocols for automated function annotation of protein structures have in many instances proven superior to simpler sequence based procedures. Here we present an automated method for extracting features from protein structures by construction of surface patches to be used in such protocols. The utility of the developed patch-growing procedure is exemplified by its ability to identify reversible membrane binding domains from the C1, C2, and PH families.

  11. Lactic acid fermentation of crude sorghum extract

    SciTech Connect

    Samuel, W.A.; Lee, Y.Y.; Anthony, W.B.

    1980-04-01

    Crude extract from sweet sorghum supplemented with vetch juice was utilized as the carbohydrate source for fermentative production of lactic acid. Fermentation of media containing 7% (w/v) total sugar was completed in 60-80 hours by Lactobacillus plantarum, product yield averaging 85%. Maximum acid production rates were dependent on pH, initial substrate distribution, and concentration, the rates varying from 2 to 5 g/liter per hour. Under limited medium supplementation the lactic acid yield was lowered to 67%. The fermented ammoniated product contained over eight times as much equivalent crude protein (N x 6.25) as the original medium. Unstructured kinetic models were developed for cell growth, lactic acid formation, and substrate consumption in batch fermentation. With the provision of experimentally determined kinetic parameters, the proposed models accurately described the fermentation process. 15 references.

  12. Comparison between modified DNA extraction protocols and commercial isolation kits in grapevine (Vitis vinifera L.).

    PubMed

    Akkurt, M

    2012-08-13

    Various protocols have been developed and used for DNA extraction in grapevine. However, owing to the long duration of the isolation steps in previously developed protocols, researchers have preferred to use isolation kits for studies in recent years. In our study, the DNA yield and purity obtained using six methods--namely three DNA isolation protocols and three commercial DNA isolation kits--were compared. Modifications were made and the isolation steps were shortened in the previously developed DNA isolation protocols to achieve more rapid and practical protocols. The samples were taken from plants grown under vineyard and greenhouse conditions in two periods during spring and autumn. The best results among the six DNA isolation methods were discussed. The results were also supported with polymerase chain reaction analyses conducted with isolated DNAs.

  13. Bacterial and fungal DNA extraction from blood samples: manual protocols.

    PubMed

    Lorenz, Michael G; Mühl, Helge; Disqué, Claudia

    2015-01-01

    A critical point of molecular diagnosis of systemic infections is the method employed for the extraction of microbial DNA from blood. A DNA isolation method has to be able to fulfill several fundamental requirements for optimal performance of diagnostic assays. First of all, low- and high-molecular-weight substances of the blood inhibitory to downstream analytical reactions like PCR amplification have to be removed. This includes human DNA which is a known source of false-positive results and factor decreasing the analytical sensitivity of PCR assays by unspecific primer binding. At the same time, even extremely low amounts of microbial DNA need to be supplied to molecular diagnostic assays in order to detect low pathogen loads in the blood. Further, considering the variety of microbial etiologies of sepsis, a method should be capable of lysing Gram-positive, Gram-negative, and fungal organisms. Last, extraction buffers, reagents, and consumables have to be free of microbial DNA which leads to false-positive results. Here, we describe manual methods which allow the extraction of microbial DNA from small- and large-volume blood samples for the direct molecular analysis of pathogen.

  14. Assessment of a sequential extraction protocol by examining solution chemistry and mineralogical evolution

    NASA Astrophysics Data System (ADS)

    Maubec, Nicolas; Pauwels, Hélène; Noël, Hervé; Bourrat, Xavier

    2015-04-01

    of them are able to leach several solid phases. In this context, the aim of the present study is to investigate the effectiveness and the selectivity of different reagents for metal extraction from target geochemical fraction. It is based on solid analyses with the use of X-ray diffraction and a scanning electron microscopy (SEM) coupled to a microRaman spectrometer in conjunction with chemical analyses of extracting solutions at each step. This methodology provides the opportunity to assess more accurately the effect of each reagent. The study focuses on extraction of Cu and Zn from sediment samples collected at two sites from river banks and characterized by presence of Quartz, Feldspar K, Micas, Kaolinite but with differences regarding accessory phases (pyrite, organic matter, iron oxy- hydroxide, calcite). The interaction of the samples with eight different reagents was assessed and compared (Ca(NO3)2 and CaCl2 for the exchangeable fraction; buffered solutions of sodium acetate/acetic acid at pH = 5.5 and pH = 5 for the acido-soluble fraction; hydroxylamine hydrochloride and a solution of ammonium oxalate/oxalic acid for reducible fraction; hydrogen peroxide and sodium hypochlorite for the oxidizable fraction. In-depth characterization of solid residue at each step allowed proposing the best protocol for both metals. Anderson, P., Davidson, C. M., Duncan, A. L., Littlejohn, D., Ure, A. M., and Garden, L. M. (2000). Column leaching and sorption experiments to assess the mobility of potentially toxic elements in industrially contaminated land. Journal of Environmental Monitoring, 2. Arey, J. S., Seaman, J. C., and Bertsch, P. M. (1999). Immobilization of uranium in contaminated sediments by hydroxyapatite addition. Environmental Science & Technology, 33, 337-342. Brannon, J. M., and Patrick, W. H. (1987). Fixation, transformation, and mobilization of arsenic in sediments.Environmental Science & Technology, 21, 450-459. Cornu, S., and Clozel, B. (2000). Extractions

  15. Investigation of aggregation in solvent extraction of lanthanides by acidic extractants (organophosphorus and naphthenic acid)

    USGS Publications Warehouse

    Zhou, N.; Wu, J.; Yu, Z.; Neuman, R.D.; Wang, D.; Xu, G.

    1997-01-01

    Three acidic extractants (I) di(2-ethylhexyl) phosphoric acid (HDEHP), (II) 2-ethylhexyl phosphonic acid mono-2-ethylhexyl ester (HEHPEHE) and (III) naphthenic acid were employed in preparing the samples for the characterization of the coordination structure of lanthanide-extractant complexes and the physicochemical nature of aggregates formed in the organic diluent of the solvent extraction systems. Photo correlation spectroscopy (PCS) results on the aggregates formed by the partially saponified HDEHP in n-heptane showed that the hydrodynamic radius of the aggregates was comparable to the molecular dimensions of HDEHP. The addition of 2-octanol into the diluent, by which the mixed solvent was formed, increased the dimensions of the corresponding aggregates. Aggregates formed from the lanthanide ions and HDEHP in the organic phase of the extraction systems were found very unstable. In the case of naphthenic acid, PCS data showed the formation of w/o microemulsion from the saponified naphthenic acid in the mixed solvent. The extraction of lanthanides by the saponified naphthenic acid in the mixed solvent under the given experimental conditions was a process of destruction of the w/o microemulsion. A possible mechanism of the breakdown of the w/o microemulsion droplets is discussed.

  16. A high-performance protocol for extraction of microplastics in fish.

    PubMed

    Karami, Ali; Golieskardi, Abolfazl; Choo, Cheng Keong; Romano, Nicholas; Ho, Yu Bin; Salamatinia, Babak

    2017-02-01

    So far, several classes of digesting solutions have been employed to extract microplastics (MPs) from biological matrices. However, the performance of digesting solutions across different temperatures has never been systematically investigated. In the first phase of the present study, we measured the efficiency of different oxidative agents (NaClO or H2O2), bases (NaOH or KOH), and acids [HCl or HNO3; concentrated and diluted (5%)] in digesting fish tissues at room temperature (RT, 25°C), 40, 50, or 60°C. In the second phase, the treatments that were efficient in digesting the biological materials (>95%) were evaluated for their compatibility with eight major plastic polymers (assessed through recovery rate, Raman spectroscopy analysis, and morphological changes). Among the tested solutions, NaClO, NaOH, and diluted acids did not result in a satisfactory digestion efficiency at any of the temperatures. The H2O2 treatment at 50°C efficiently digested the biological materials, although it decreased the recovery rate of nylon-6 (NY6) and nylon-66 (NY66) and altered the colour of polyethylene terephthalate (PET) fragments. Similarly, concentrated HCl and HNO3 treatments at RT fully digested the fish tissues, but also fully dissolved NY6 and NY66, and reduced the recovery rate of most or all of the polymers, respectively. Potassium hydroxide solution fully eliminated the biological matrices at all temperatures. However, at 50 and 60°C, it degraded PET, reduced the recovery rate of PET and polyvinyl chloride (PVC), and changed the colour of NY66. According to our results, treating biological materials with a 10% KOH solution and incubating at 40°C was both time and cost-effective, efficient in digesting biological materials, and had no impact on the integrity of the plastic polymers. Furthermore, coupling this treatment with NaI extraction created a promising protocol to isolate MPs from whole fish samples.

  17. A "novel" protocol for the analysis of hydroxycinnamic acids in leaf tissue of chicory (Cichorium intybus L., Asteraceae).

    PubMed

    Bahri, Meriem; Hance, Philippe; Grec, Sébastien; Quillet, Marie-Christine; Trotin, Francis; Hilbert, Jean-Louis; Hendriks, Theo

    2012-01-01

    A "novel" protocol is presented for easy and reliable estimation of soluble hydroxycinnamate levels in Cichorium intybus L. leaf tissue in large-scale experiments. Samples were standardized by punching 6 discs per leaf, and hydroxycinnamates were extracted by submerging the discs in 80% ethanol with 5% acetic acid for at least 48 h in the darkness at 4°C. Residual dry mass of the discs was used for a posteriori correction of compound levels. Chlorophyll was eliminated by chloroform, and the aqueous phases were transferred to microplates, dried, and dissolved in 50% methanol for HPLC analysis and storage. An HPLC program of 8 min was developed for the analysis of the extracts. Comparisons with extractions of liquid nitrogen powders indicated that the novel extraction method was reliable. No degradation of the major hydroxycinnamates-caftaric, chlorogenic, and chicoric acids-was observed, during maceration at ambient temperatures, or after storage for 1 year.

  18. Comparison of extraction protocols to determine differences in wine-extractable tannin and anthocyanin in Vitis vinifera L. cv. Shiraz and Cabernet Sauvignon grapes.

    PubMed

    Bindon, Keren A; Kassara, Stella; Cynkar, Wieslawa U; Robinson, Ella M C; Scrimgeour, Neil; Smith, Paul A

    2014-05-21

    Cabernet Sauvignon and Shiraz grapes were sourced from different regions within Australia, and microvinified with a skin contact period of 6 days. Grape samples were extracted using two protocols: a 15% v/v ethanol, 10 g/L tartaric acid extract of gently crushed berries (wine-like, WL) and a 50% v/v ethanol, pH 2 extract of grape berry homogenate. It was found that in WL extracts, grape tannin and anthocyanin concentrations were strongly related to wine tannin, anthocyanin and color density achieved during the skin contact period. No relationship was observed for grape tannin concentration analyzed in homogenate extracts and wine tannin, but a strong, positive relationship was found for anthocyanin concentration. When the data obtained from homogenate extraction was treated separately by grape variety, a stronger relationship between grape and wine tannin concentration was observed. Tannin compositional analysis in wines indicated that higher tannin concentrations were due to the extraction of tannin of higher molecular mass during fermentation, most likely from grape skins.

  19. A modified extraction protocol enables detection and quantification of celiac disease-related gluten proteins from wheat.

    PubMed

    van den Broeck, Hetty C; America, Antoine H P; Smulders, Marinus J M; Bosch, Dirk; Hamer, Rob J; Gilissen, Ludovicus J W J; van der Meer, Ingrid M

    2009-04-01

    The detection, analysis, and quantification of individual celiac disease (CD) immune responsive gluten proteins in wheat and related cereals (barley, rye) require an adequate and reliable extraction protocol. Because different types of gluten proteins behave differently in terms of solubility, currently different extraction protocols exist. The performance of various documented gluten extraction protocols is evaluated for specificity and completeness by gel electrophoresis (SDS-PAGE), immunoblotting and RIDASCREEN Gliadin competitive ELISA. Based on these results, an optimized, two-step extraction protocol has been developed.

  20. An improved protocol for DNA extraction from alkaline soil and sediment samples for constructing metagenomic libraries.

    PubMed

    Verma, Digvijay; Satyanarayana, T

    2011-09-01

    An improved single-step protocol has been developed for extracting pure community humic substance-free DNA from alkaline soils and sediments. The method is based on direct cell lysis in the presence of powdered activated charcoal and polyvinylpolypyrrolidone followed by precipitation with polyethyleneglycol and isopropanol. The strategy allows simultaneous isolation and purification of DNA while minimizing the loss of DNA with respect to other available protocols for metagenomic DNA extraction. Moreover, the purity levels are significant, which are difficult to attain with any of the methods reported in the literature for DNA extraction from soils. The DNA thus extracted was free from humic substances and, therefore, could be processed for restriction digestion, PCR amplification as well as for the construction of metagenomic libraries.

  1. A High-Yield Two-Hour Protocol for Extraction of Human Hair Shaft Proteins

    PubMed Central

    Wong, Sing Ying; Lee, Ching Chin; Ashrafzadeh, Ali; Junit, Sarni Mat; Abrahim, Nazirahanie

    2016-01-01

    Proteome analysis of the human hair remains challenging due to the poor solubility of hair proteins and the difficulty in their extraction. In the present study, we have developed a rapid extraction protocol for hair shaft protein using alkaline-based buffer. The new protocol accelerated the procedure by reducing the extraction time from at least a day to less than two hours and showed a protein recovery of 47.3 ± 3.72%. Further analyses of the extracted protein sample through sodium dodecyl sulfate polyacrylamide gel electrophoresis and Quadrupole-time-of-flight mass spectrometry analysis unveiled a total of 60 proteins, including 25 that were not previously reported. Identification of these proteins is anticipated to be crucial in helping to understand the molecular basis of hair for potential applications in the future. PMID:27741315

  2. Histone extraction protocol from the two model diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana.

    PubMed

    Tirichine, Leïla; Lin, Xin; Thomas, Yann; Lombard, Bérangère; Loew, Damarys; Bowler, Chris

    2014-02-01

    Post-translational modifications of histones affect many biological processes by influencing higher order chromatin structure that affects gene and genome regulation. It is therefore important to develop methods for extracting histones while maintaining their native post-translational modifications. While histone extraction protocols have been developed in multicellular and single celled organisms such as yeast and Arabidopsis, they are inefficient in diatoms that have a silica cell wall that is likely to hinder histone extraction. We report in this work a rapid and reliable method for extraction of large amounts of high quality histones from the two model diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. The protocol is an important enabling step permitting downstream applications such as western blotting and mass spectrometry.

  3. A High-Yield Two-Hour Protocol for Extraction of Human Hair Shaft Proteins.

    PubMed

    Wong, Sing Ying; Lee, Ching Chin; Ashrafzadeh, Ali; Junit, Sarni Mat; Abrahim, Nazirahanie; Hashim, Onn Haji

    2016-01-01

    Proteome analysis of the human hair remains challenging due to the poor solubility of hair proteins and the difficulty in their extraction. In the present study, we have developed a rapid extraction protocol for hair shaft protein using alkaline-based buffer. The new protocol accelerated the procedure by reducing the extraction time from at least a day to less than two hours and showed a protein recovery of 47.3 ± 3.72%. Further analyses of the extracted protein sample through sodium dodecyl sulfate polyacrylamide gel electrophoresis and Quadrupole-time-of-flight mass spectrometry analysis unveiled a total of 60 proteins, including 25 that were not previously reported. Identification of these proteins is anticipated to be crucial in helping to understand the molecular basis of hair for potential applications in the future.

  4. Evaluation of four automated protocols for extraction of DNA from FTA cards.

    PubMed

    Stangegaard, Michael; Børsting, Claus; Ferrero-Miliani, Laura; Frank-Hansen, Rune; Poulsen, Lena; Hansen, Anders J; Morling, Niels

    2013-10-01

    Extraction of DNA using magnetic bead-based techniques on automated DNA extraction instruments provides a fast, reliable, and reproducible method for DNA extraction from various matrices. Here, we have compared the yield and quality of DNA extracted from FTA cards using four automated extraction protocols on three different instruments. The extraction processes were repeated up to six times with the same pieces of FTA cards. The sample material on the FTA cards was either blood or buccal cells. With the QIAamp DNA Investigator and QIAsymphony DNA Investigator kits, it was possible to extract DNA from the FTA cards in all six rounds of extractions in sufficient amount and quality to obtain complete short tandem repeat (STR) profiles on a QIAcube and a QIAsymphony SP. With the PrepFiler Express kit, almost all the extractable DNA was extracted in the first two rounds of extractions. Furthermore, we demonstrated that it was possible to successfully extract sufficient DNA for STR profiling from previously processed FTA card pieces that had been stored at 4 °C for up to 1 year. This showed that rare or precious FTA card samples may be saved for future analyses even though some DNA was already extracted from the FTA cards.

  5. Process for the extraction of strontium from acidic solutions

    DOEpatents

    Horwitz, E.P.; Dietz, M.L.

    1993-01-01

    The invention is a process for selectively extracting strontium values from aqueous nitric acid waste solutions containing these and other fission product values. The extractant solution is a macrocyclic polyether in an aliphatic hydrocarbon diluent containing a phase modifier. The process will selectively extract strontium values from nitric acid solutions which are up to 6 molar in nitric acid.

  6. Process for the extraction of strontium from acidic solutions

    DOEpatents

    Horwitz, E.P.; Dietz, M.L.

    1994-09-06

    The invention is a process for selectively extracting strontium values from aqueous nitric acid waste solutions containing these and other fission product values. The extractant solution is a macrocyclic polyether in an aliphatic hydrocarbon diluent containing a phase modifier. The process will selectively extract strontium values from nitric acid solutions which are up to 6 molar in nitric acid. 4 figs.

  7. Process for the extraction of strontium from acidic solutions

    DOEpatents

    Horwitz, E. Philip; Dietz, Mark L.

    1994-01-01

    The invention is a process for selectively extracting strontium values from aqueous nitric acid waste solutions containing these and other fission product values. The extractant solution is a macrocyclic polyether in an aliphatic hydrocarbon diluent containing a phase modifier. The process will selectively extract strontium values from nitric acid solutions which are up to 6 molar in nitric acid.

  8. Treatment of Class III Malocclusion: Atypical Extraction Protocol

    PubMed Central

    Ferreira, Fernando Pedrin Carvalho; de Almeida-Pedrin, Renata Rodrigues; Conti, Ana Claudia de Castro Ferreira; Cardoso, Maurício de Almeida

    2017-01-01

    The treatment of Angle Class III malocclusion is rather challenging, because the patient's growth pattern determines the success of long-term treatment. Early diagnosis and treatment are still highly discussed issues in orthodontic literature. This type of early intervention has been indicated more frequently in order to eliminate primary etiological factors and prevent an already present malocclusion from becoming severe. However, when a patient is diagnosed in adulthood, manipulation of the bone bases becomes extremely limited, as there is no longer any potential for growth. Treatments are restricted to dental compensations when possible or orthognathic surgery. However, owing to the high cost and inherent risk of the surgical procedure, this treatment option is often denied by the patient; in such a case, the orthodontist has little choice but to perform, where possible, compensatory treatments to restore a functional occlusion and improve facial esthetics. This article reports a case of Class III malocclusion in a patient who opted for compensatory treatment with lower molar extraction that allowed for correction of the midline and the overjet. Good facial esthetics and functional normal occlusion were achieved at the end of the treatment. PMID:28265473

  9. Development of extraction protocols for life detection biosensor-based instruments

    NASA Astrophysics Data System (ADS)

    Fornaro, Teresa; Brucato, John Robert; Pucci, Amaranta; Branciamore, Sergio

    2013-09-01

    Extraction experiments were performed to evaluate suitable sample-extraction and processing protocols that will be used by bioanalytical instruments like Life Marker Chip (LMC). These instruments will focus on the detection of molecules associated with life that will be extracted from the Martian soil. LMC is an antibody microarray biosensor instrument with optical readout, which uses fluorescently labeled antibodies, to detect and quantify the presence of polar and non-polar biomolecules, extracted from the Martian soil. The success of the LMC biomarkers detection experiment is dependent on the sample extraction protocol. The currently recommended extraction procedure, optimized for aliphatic biomarkers, consists in sonicating the samples using as solvent system 20:80 methanol:water with 1.5 mg mL-1 tween 80, which respects all the tight requirements associated with LMC biosensor. Examining this procedure for a particular class of biomarkers, the nucleobases adenine, cytosine, uracil and hypoxanthine adsorbed onto magnesium oxide (MgO), we observed low extraction efficiency, confirming that the selected solvent system is not able to extract with high efficiency aromatic biomarkers.

  10. A Reliable and Inexpensive Method of Nucleic Acid Extraction for the PCR-Based Detection of Diverse Plant Pathogens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A reliable extraction method is described for the preparation of total nucleic acids from several plant genera for subsequent detection of plant pathogens by PCR-based techniques. By the combined use of a modified CTAB (cetyltrimethylammonium bromide) extraction protocol and a semi-automatic homogen...

  11. A protocol for protein extraction from lipid-rich plant tissues suitable for electrophoresis.

    PubMed

    Zienkiewicz, Agnieszka; Rejón, Juan David; de Dios Alché, Juan; Rodríguez-García, María Isabel; Castro, Antonio Jesús

    2014-01-01

    Plant tissues contain high levels of nonprotein contaminants such as lipids, phenolic compounds, and polysaccharides among others, which interfere with protein extraction and electrophoretic separation. Preparation of good-quality protein extracts is a critical issue for successful electrophoretic analysis. Here, we describe a three-step method for protein extraction from lipid-rich plant tissues, which is suitable for both 1-D and 2-D electrophoresis and is compatible with downstream applications. The protocol includes prefractionation, filtration, and TCA/acetone precipitation steps prior to protein resolubilization.

  12. Fast protocol for extraction of DNA from Prosopis spp leaves (plant adapted to arid environment) without liquid nitrogen.

    PubMed

    Michel-López, C Y; González-Mendoza, D; Grimaldo-Juarez, O

    2013-09-27

    The extraction of high-quality genomic DNA from Prosopis spp for polymerase chain reaction (PCR) amplification is complicated, owing to the presence of a high percentage of secondary metabolites that bind to or co-precipitate with nucleic acids. In the present study, we report a modified sodium dodecyl sulfate/phenol protocol that eliminates the use of liquid nitrogen in the maceration process, β-mercaptoethanol in the buffer extraction, and the ethanol precipitation step. The A₂₆₀/A₂₈₀ absorbance ratios of the isolated DNA were approximately 2.0 to 1.9, suggesting that the DNA fraction was pure and can be used for further PCR analysis. The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction. Finally, this proposal represents an alternative fast, cheap, and effective method for the isolation of genomic DNA from fresh leaves of Prosopis spp, even in low-technology laboratories.

  13. Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays.

    PubMed

    Chen, Qingwen; Narayanan, Kumaran

    2011-07-01

    The phage N15 protelomerase enzyme (TelN) is essential for the replication of its genome by resolution of its telRL domain, located within a telomerase occupancy site (tos), into hairpin telomeres. Isolation of TelN for in vitro processing of tos, however, is a highly complex process, requiring multiple purification steps. In this study a simplified protocol for crude total protein extraction is described that retains the tos-cleaving activity of TelN for at least 4 weeks, greatly simplifying in vitro testing of its activity. This protocol may be extended for functional analysis of other phage and bacterial proteins, particularly DNA-processing enzymes.

  14. Safety of dental extraction among consecutive patients on oral anticoagulant treatment managed using a specific dental management protocol.

    PubMed

    Zanon, Ezio; Martinelli, Franco; Bacci, Christian; Cordioli, GianPiero; Girolami, Antonio

    2003-01-01

    We found no prospective studies on dental extraction in anticoagulated patients in the literature, even though most authors suggest that there is no need to change anticoagulant treatment and to utilize a local haemostatic measure after extraction. In the present study, we have verified the incidence of bleeding complications after dental extraction in a group of 250 consecutive anticoagulated patients. Two hundred and fifty non-anticoagulated subjects requiring dental extraction represented the control group. In all patients, anticoagulant treatment was not changed (International Normalized Ratio, 1.8-4) and local haemostatic measures (fibrin sponge, silk suture and a gauze saturated with tranexamic acid) were used. All procedures were performed in an outpatient clinic setting. We registered four bleeding complications in the group of anticoagulated patients and three in the control group. The difference of bleeding complications in the two groups was not statistically different (relative risk, 1.14; 95% confidence interval, 0.29-6.04; P = 0.7). None of the post-operative late bleeding required hospitalization and/or blood transfusions, and further local measures were sufficient to stop the bleeding. The protocol proposed in the present study makes dental extractions in anticoagulated patients possible on an outpatient basis with a cost reduction for the community and minor discomfort for the patients.

  15. Extraction of fatty acids from dried freshwater algae using accelerated solvent extraction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A high temperature/pressure extraction method (accelerated solvent extraction)(ASE) and a manual extraction method (modified Folch extraction) were compared with regard to their ability to extract total fat from three samples of air-dried filamentous algae and determine the fatty acid (FA) profile o...

  16. Modified CTAB and TRIzol protocols improve RNA extraction from chemically complex Embryophyta1

    PubMed Central

    Jordon-Thaden, Ingrid E.; Chanderbali, Andre S.; Gitzendanner, Matthew A.; Soltis, Douglas E.

    2015-01-01

    Premise of the study: Here we present a series of protocols for RNA extraction across a diverse array of plants; we focus on woody, aromatic, aquatic, and other chemically complex taxa. Methods and Results: Ninety-one taxa were subjected to RNA extraction with three methods presented here: (1) TRIzol/TURBO DNA-free kits using the manufacturer’s protocol with the addition of sarkosyl; (2) a combination method using cetyltrimethylammonium bromide (CTAB) and TRIzol/sarkosyl/TURBO DNA-free; and (3) a combination of CTAB and QIAGEN RNeasy Plant Mini Kit. Bench-ready protocols are given. Conclusions: After an iterative process of working with chemically complex taxa, we conclude that the use of TRIzol supplemented with sarkosyl and the TURBO DNA-free kit is an effective, efficient, and robust method for obtaining RNA from 100 mg of leaf tissue of land plant species (Embryophyta) examined. Our protocols can be used to provide RNA of suitable stability, quantity, and quality for transcriptome sequencing. PMID:25995975

  17. Astrobionibbler: In Situ Microfluidic Subcritical Water Extraction of Amino Acids

    NASA Astrophysics Data System (ADS)

    Noell, A. C.; Fisher, A. M.; Takano, N.; Fors-Francis, K.; Sherrit, S.; Grunthaner, F.

    2016-10-01

    A fluidic-chip based instrument for subcritical water extraction (SCWE) of amino acids and other organics from powder samples has been developed. A variety of soil analog extractions have been performed to better understand SCWE capabilities.

  18. DNA extraction from bristles and quills of Chaetomys subspinosus (Rodentia: Erethizontidae) using a novel protocol.

    PubMed

    Oliveira, C G; Martinez, R A; Gaiotto, F A

    2007-09-30

    DNA extraction protocols are as varied as DNA sources. When it comes to endangered species, it is especially important to pay attention to all details that ensure the completion of the study goals and effectiveness in attaining useful data for conservation. Chaetomys subspinosus (Rodentia: Erethizontidae) is a secretive arboreal porcupine endemic to certain ecosystems of the Brazilian Atlantic Forest. A multidisciplinary study (including genetic data) was performed to create a management plan for the conservation of this species. Individuals from natural populations of the states of Bahia, Espírito Santo and Sergipe were sampled. To obtain a reliable and abundant amount of starting material, non-destructive methods were tested, extracting DNA from the bristles and quills that comprise most of this animal's hide. This method has also been innovative in adapting a DNA extraction protocol traditionally used for plants. Digestion using proteinase K was followed by protein precipitation with CTAB, a chloroform-isoamyl alcohol cleaning and DNA precipitation with isopropyl alcohol. This protocol supplies good-quality DNA for genetic analysis with molecular markers based on PCR.

  19. Recovery of boric acid from wastewater by solvent extraction

    SciTech Connect

    Matsumoto, Michiaki; Kondo, Kazuo; Hirata, Makoto; Kokubu, Shuzo; Hano, Tadashi

    1997-03-01

    An extraction system for the recovery of boric acid using 2-butyl-2-ethyl-1,3-propanediol (BEPD) as an extractant was studied. Loss of the extractant to the aqueous solution was lowered by using 2-ethylhexanol as a diluent. The extraction equilibrium of boric acid with BEPD was clarified, and the equilibrium constants for various diluents were determined. Furthermore, continuous operation for the recovery of boric acid using mixer-settlers for extraction and stripping was successfully conducted during 100 hours. 10 refs., 5 figs., 2 tabs.

  20. Strategic extraction protocol: Use of an image-fusion stereolithographic guide for immediate implant placement.

    PubMed

    Dada, Karim; Pariente, Leon; Daas, Marwan

    2016-11-01

    Increasingly, patients are demanding a smooth transition from a hopeless dentition to a fixed implant-supported prosthesis without the use of an interim removable denture. This raises new challenges in adapting computer-aided design and computer-aided manufacturing (CAD-CAM) techniques to patients whose treatment plan includes immediate implant placement in fresh extraction sockets. To facilitate this conversion to a rehabilitated dentition, a strategic extraction protocol (SEP) can be applied. The SEP consists of selecting strategic, remaining, hopeless teeth as abutments to stabilize an image-fusion stereolithographic guide and to allow the immediate placement of sufficient implants to support an immediately loaded fixed prosthesis. The remaining supporting teeth are extracted in the same surgical session before the placement of the interim prosthesis.

  1. A protocol for high-quality genomic DNA extraction from legumes.

    PubMed

    Agbagwa, I O; Datta, S; Patil, P G; Singh, P; Nadarajan, N

    2012-12-19

    Current DNA extraction protocols, which require liquid nitrogen, lyophilization and considerable infrastructure in terms of instrumentation, often impede the application of biotechnological tools in less researched crops in laboratories in developing countries. We modified and optimized the existing CTAB method for plant genomic DNA extraction by avoiding liquid nitrogen usage and lyophilization. DNA was extracted directly from freshly harvested leaves ground in pre-heated CTAB buffer. Chloroform:isoamyl alcohol (24:1) and RNase treatments followed by single-purification step decontaminated the samples thereby paving way for selective extraction of DNA. High molecular weight DNA yield in the range of 328 to 4776 ng/μL with an average of 1459 ng/μL was obtained from 45 samples of cultivated and wild Cajanus species. With an absorbance ratio at 260 to 280 nm, a range of 1.66 to 2.20, and a mean of 1.85, very low levels of protein and polysaccharide contamination were recorded. Forty samples can be extracted daily at a cost between 1.8 and US$2.0 per plant sample. This modified method is suitable for most plants especially members of the Leguminosae. Apart from Cajanus, it has been extensively applied in DNA extraction from Cicer and Vigna species.

  2. Comparison of different protocols for the extraction of microbial DNA from reef corals

    PubMed Central

    Santos, H.F.; Carmo, F.L.; Leite, D.C.A.; Jesus, H.E.; Maalouf, P. De Carvalho; Almeida, C.; Soriano, A.U.; Altomari, D.; Suhett, L.; Vólaro, V.; Valoni, E.; Francisco, M.; Vieira, J.; Rocha, R.; Sardinha, B.L.; Mendes, L.B.; João, R.R.; Lacava, B.; Jesus, R.F.; Sebastian, G.V.; Pessoa, A.; van Elsas, J.D.; Rezende, R.P.; Pires, D.O.; Duarte, G.; Castro, C.B.; Rosado, A.S.; Peixoto, R.S.

    2012-01-01

    This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals. PMID:24031859

  3. Comparison of different protocols for the extraction of microbial DNA from reef corals.

    PubMed

    Santos, H F; Carmo, F L; Leite, D C A; Jesus, H E; Maalouf, P De Carvalho; Almeida, C; Soriano, A U; Altomari, D; Suhett, L; Vólaro, V; Valoni, E; Francisco, M; Vieira, J; Rocha, R; Sardinha, B L; Mendes, L B; João, R R; Lacava, B; Jesus, R F; Sebastian, G V; Pessoa, A; van Elsas, J D; Rezende, R P; Pires, D O; Duarte, G; Castro, C B; Rosado, A S; Peixoto, R S

    2012-04-01

    This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.

  4. DNA extraction protocol for biological ingredient analysis of Liuwei Dihuang Wan.

    PubMed

    Cheng, Xinwei; Chen, Xiaohua; Su, Xiaoquan; Zhao, Huanxin; Han, Maozhen; Bo, Cunpei; Xu, Jian; Bai, Hong; Ning, Kang

    2014-06-01

    Traditional Chinese medicine (TCM) preparations are widely used for healthcare and clinical practice. So far, the methods commonly used for quality evaluation of TCM preparations mainly focused on chemical ingredients. The biological ingredient analysis of TCM preparations is also important because TCM preparations usually contain both plant and animal ingredients, which often include some mis-identified herbal materials, adulterants or even some biological contaminants. For biological ingredient analysis, the efficiency of DNA extraction is an important factor which might affect the accuracy and reliability of identification. The component complexity in TCM preparations is high, and DNA might be destroyed or degraded in different degrees after a series of processing procedures. Therefore, it is necessary to establish an effective protocol for DNA extraction from TCM preparations. In this study, we chose a classical TCM preparation, Liuwei Dihuang Wan (LDW), as an example to develop a TCM-specific DNA extraction method. An optimized cetyl trimethyl ammonium bromide (CTAB) method (TCM-CTAB) and three commonly-used extraction kits were tested for extraction of DNA from LDW samples. Experimental results indicated that DNA with the highest purity and concentration was obtained by using TCM-CTAB. To further evaluate the different extraction methods, amplification of the second internal transcribed spacer (ITS2) and the chloroplast genome trnL intron was carried out. The results have shown that PCR amplification was successful only with template of DNA extracted by using TCM-CTAB. Moreover, we performed high-throughput 454 sequencing using DNA extracted by TCM-CTAB. Data analysis showed that 3-4 out of 6 prescribed species were detected from LDW samples, while up to 5 contaminating species were detected, suggesting TCM-CTAB method could facilitate follow-up DNA-based examination of TCM preparations.

  5. Fermentation of aqueous plant seed extracts by lactic acid bacteria

    SciTech Connect

    Schafner, D.W.; Beuchat, R.L.

    1986-05-01

    The effects of lactic acid bacterial fermentation on chemical and physical changes in aqueous extracts of cowpea (Vigna unguiculata), peanut (Arachis hypogea), soybean (Glycine max), and sorghum (Sorghum vulgare) were studied. The bacteria investigated were Lactobacillus helveticus, L. delbrueckii, L. casei, L. bulgaricus, L. acidophilus, and Streptococcus thermophilus. Organisms were inoculated individually into all of the seed extracts; L. bulgaricus and S. thermophilus were also evaluated together as inocula for fermenting the legume extracts. During fermentation, bacterial population and changes in titratable acidity, pH, viscosity, and color were measured over a 72 h period at 37 degrees C. Maximum bacterial populations, titratable acidity, pH, and viscosity varied depending upon the type of extract and bacterial strain. The maximum population of each organism was influenced by fermentable carbohydrates, which, in turn, influenced acid production and change in pH. Change in viscosity was correlated with the amount of protein and titratable acidity of products. Color was affected by pasteurization treatment and fermentation as well as the source of extract. In the extracts inoculated simultaneously with L. bulgaricus and S. thermophilus, a synergistic effect resulted in increased bacterial populations, titratable acidity, and viscosity, and decreased pH in all the legume extracts when compared to the extracts fermented with either of these organisms individually. Fermented extracts offer potential as substitutes for cultured dairy products. 24 references.

  6. Microwave-Assisted Extraction of Oleanolic Acid and Ursolic Acid from Ligustrum lucidum Ait

    PubMed Central

    Xia, En-Qin; Wang, Bo-Wei; Xu, Xiang-Rong; Zhu, Li; Song, Yang; Li, Hua-Bin

    2011-01-01

    Oleanolic acid and ursolic acid are the main active components in fruit of Ligustrum lucidum Ait, and possess anticancer, antimutagenic, anti-inflammatory, antioxidative and antiprotozoal activities. In this study, microwave-assisted extraction of oleanolic acid and ursolic acid from Ligustrum lucidum was investigated with HPLC-photodiode array detection. Effects of several experimental parameters, such as type and concentration of extraction solvent, ratio of liquid to material, microwave power, extraction temperature and microwave time, on the extraction efficiencies of oleanolic acid and ursolic acid from Ligustrum lucidum were evaluated. The influence of experimental parameters on the extraction efficiency of ursolic acid was more significant than that of oleanolic acid (p < 0.05). The optimal extraction conditions were 80% ethanol aqueous solution, the ratio of material to liquid was 1:15, and extraction for 30 min at 70 °C under microwave irradiation of 500 W. Under optimal conditions, the yields of oleanolic acid and ursolic acid were 4.4 ± 0.20 mg/g and 5.8 ± 0.15 mg/g, respectively. The results obtained are helpful for the full utilization of Ligustrum lucidum, which also indicated that microwave-assisted extraction is a very useful method for extraction of oleanolic acid and ursolic acid from plant materials. PMID:21954361

  7. A Modified Protocol for High-Quality RNA Extraction from Oleoresin-Producing Adult Pines.

    PubMed

    de Lima, Júlio César; Füller, Thanise Nogueira; de Costa, Fernanda; Rodrigues-Corrêa, Kelly C S; Fett-Neto, Arthur G

    2016-01-01

    RNA extraction resulting in good yields and quality is a fundamental step for the analyses of transcriptomes through high-throughput sequencing technologies, microarray, and also northern blots, RT-PCR, and RTqPCR. Even though many specific protocols designed for plants with high content of secondary metabolites have been developed, these are often expensive, time consuming, and not suitable for a wide range of tissues. Here we present a modification of the method previously described using the commercially available Concert™ Plant RNA Reagent (Invitrogen) buffer for field-grown adult pine trees with high oleoresin content.

  8. Comparison of commercial systems for extraction of nucleic acids from DNA/RNA respiratory pathogens.

    PubMed

    Yang, Genyan; Erdman, Dean E; Kodani, Maja; Kools, John; Bowen, Michael D; Fields, Barry S

    2011-01-01

    This study compared six automated nucleic acid extraction systems and one manual kit for their ability to recover nucleic acids from human nasal wash specimens spiked with five respiratory pathogens, representing Gram-positive bacteria (Streptococcus pyogenes), Gram-negative bacteria (Legionella pneumophila), DNA viruses (adenovirus), segmented RNA viruses (human influenza virus A), and non-segmented RNA viruses (respiratory syncytial virus). The robots and kit evaluated represent major commercially available methods that are capable of simultaneous extraction of DNA and RNA from respiratory specimens, and included platforms based on magnetic-bead technology (KingFisher mL, Biorobot EZ1, easyMAG, KingFisher Flex, and MagNA Pure Compact) or glass fiber filter technology (Biorobot MDX and the manual kit Allprep). All methods yielded extracts free of cross-contamination and RT-PCR inhibition. All automated systems recovered L. pneumophila and adenovirus DNA equivalently. However, the MagNA Pure protocol demonstrated more than 4-fold higher DNA recovery from the S. pyogenes than other methods. The KingFisher mL and easyMAG protocols provided 1- to 3-log wider linearity and extracted 3- to 4-fold more RNA from the human influenza virus and respiratory syncytial virus. These findings suggest that systems differed in nucleic acid recovery, reproducibility, and linearity in a pathogen specific manner.

  9. Protocols for nuclei isolation and nuclear protein extraction from the resurrection plant Xerophyta viscosa for proteomic studies.

    PubMed

    Abdalla, Kamal Omer; Thomson, Jennifer Ann; Rafudeen, Muhammad Suhail

    2009-01-15

    The plant nucleus is an important subcellular organelle but the isolation of pure and enriched nuclei from plants and subsequent extraction of nuclear proteins for proteomic studies is challenging. Here, we present protocols for nuclei isolation and nuclear protein extraction from the resurrection plant, Xerophyta viscosa, and show optimization and modification of the most critical steps.

  10. An economical and effective high-throughput DNA extraction protocol for molecular marker analysis in honey bees

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Extraction of DNA from tissue samples can be expensive both in time and monetary resources and can often require handling and disposal of hazardous chemicals. We have developed a high throughput protocol for extracting DNA from honey bees that is of a high enough quality and quantity to enable hundr...

  11. Streamlining DNA Barcoding Protocols: Automated DNA Extraction and a New cox1 Primer in Arachnid Systematics

    PubMed Central

    Vidergar, Nina; Toplak, Nataša; Kuntner, Matjaž

    2014-01-01

    Background DNA barcoding is a popular tool in taxonomic and phylogenetic studies, but for most animal lineages protocols for obtaining the barcoding sequences—mitochondrial cytochrome C oxidase subunit I (cox1 AKA CO1)—are not standardized. Our aim was to explore an optimal strategy for arachnids, focusing on the species-richest lineage, spiders by (1) improving an automated DNA extraction protocol, (2) testing the performance of commonly used primer combinations, and (3) developing a new cox1 primer suitable for more efficient alignment and phylogenetic analyses. Methodology We used exemplars of 15 species from all major spider clades, processed a range of spider tissues of varying size and quality, optimized genomic DNA extraction using the MagMAX Express magnetic particle processor—an automated high throughput DNA extraction system—and tested cox1 amplification protocols emphasizing the standard barcoding region using ten routinely employed primer pairs. Results The best results were obtained with the commonly used Folmer primers (LCO1490/HCO2198) that capture the standard barcode region, and with the C1-J-2183/C1-N-2776 primer pair that amplifies its extension. However, C1-J-2183 is designed too close to HCO2198 for well-interpreted, continuous sequence data, and in practice the resulting sequences from the two primer pairs rarely overlap. We therefore designed a new forward primer C1-J-2123 60 base pairs upstream of the C1-J-2183 binding site. The success rate of this new primer (93%) matched that of C1-J-2183. Conclusions The use of C1-J-2123 allows full, indel-free overlap of sequences obtained with the standard Folmer primers and with C1-J-2123 primer pair. Our preliminary tests suggest that in addition to spiders, C1-J-2123 will also perform in other arachnids and several other invertebrates. We provide optimal PCR protocols for these primer sets, and recommend using them for systematic efforts beyond DNA barcoding. PMID:25415202

  12. Comparative analysis of different DNA extraction protocols in fresh and herbarium specimens of the genus Dalbergia.

    PubMed

    Ribeiro, R A; Lovato, M B

    2007-03-29

    Five published DNA extraction protocols were compared for their ability to produce good quality DNA from fresh and herbarium leaves of several species of the genus Dalbergia. The leaves of these species contain high amounts of secondary metabolites, which make it difficult to perform a clean DNA extraction and thereby interfering with subsequent PCR amplification. The protocol that produced the best DNA quality in most of the Dalbergia species analyzed, utilizes polyvinylpyrrolidone to bind the phenolic compounds, a high molar concentration of NaCl to inhibit co-precipitation of polysaccharides and DNA, and LiCl for removing RNA by selective precipitation. The DNA quality of herbarium specimens was worse than that for fresh leaves, due to collecting conditions and preservation of samples. We analyzed 54 herbarium specimens, but the recovered DNA allowed successful PCR amplification in only eight. For the genus Dalbergia, the herbarium is an important source of material for phylogenetic and evolutionary studies; due to the occurrence of the different species in various geographical regions in Brazil, it is difficult to obtain fresh material in nature. Our results demonstrated that for Dalbergia species the methods used for the collection and preservation of herbarium specimens have a mayor influence on DNA quality and in the success of phylogenetic studies of the species.

  13. DNA Extraction Protocols for Whole-Genome Sequencing in Marine Organisms.

    PubMed

    Panova, Marina; Aronsson, Henrik; Cameron, R Andrew; Dahl, Peter; Godhe, Anna; Lind, Ulrika; Ortega-Martinez, Olga; Pereyra, Ricardo; Tesson, Sylvie V M; Wrange, Anna-Lisa; Blomberg, Anders; Johannesson, Kerstin

    2016-01-01

    The marine environment harbors a large proportion of the total biodiversity on this planet, including the majority of the earths' different phyla and classes. Studying the genomes of marine organisms can bring interesting insights into genome evolution. Today, almost all marine organismal groups are understudied with respect to their genomes. One potential reason is that extraction of high-quality DNA in sufficient amounts is challenging for many marine species. This is due to high polysaccharide content, polyphenols and other secondary metabolites that will inhibit downstream DNA library preparations. Consequently, protocols developed for vertebrates and plants do not always perform well for invertebrates and algae. In addition, many marine species have large population sizes and, as a consequence, highly variable genomes. Thus, to facilitate the sequence read assembly process during genome sequencing, it is desirable to obtain enough DNA from a single individual, which is a challenge in many species of invertebrates and algae. Here, we present DNA extraction protocols for seven marine species (four invertebrates, two algae, and a marine yeast), optimized to provide sufficient DNA quality and yield for de novo genome sequencing projects.

  14. Microwave-assisted extraction versus Soxhlet extraction to determine triterpene acids in olive skins.

    PubMed

    Fernandez-Pastor, Ignacio; Fernandez-Hernandez, Antonia; Perez-Criado, Sergio; Rivas, Francisco; Martinez, Antonio; Garcia-Granados, Andres; Parra, Andres

    2017-03-01

    Microwave-assisted extraction is compared with a more classical technique, Soxhlet extraction, to determine the content of triterpene acids in olive skins. The samples used in their original unmilled state and milled were extracted with ethyl acetate or methanol as solvents. The optimized operating conditions (e.g., amount and type of solvent, and time and temperature of extractions) to attain the better extraction yields have been established. For the identification and quantitation of the target compounds, an ultra high performance liquid chromatography with tandem mass spectrometry method was employed. The best results were achieved using the microwave-assisted extraction technique, which was much faster than the Soxhlet extraction method, and showed higher efficiency in the extraction of the triterpenic acids (oleanolic and maslinic).

  15. Solvent extraction of rare-earth metals by carboxylic acids

    SciTech Connect

    Preez, A.C. du; Preston, J.S.

    1992-04-01

    The solvent extraction of the trivalent lanthanides and yttrium from nitrate media by solutions of carboxylic acids in xylene has been studied. Commercially available carboxylic acids such as Versatic 10 and naphthenic acids were used, as well as model compounds of known structure, such as 2-ethylhexanoic and 3-cyclohexylpropanoic acids. In a few cases, extraction of the metals from sulphate and chloride solutions was also investigated. The dependence of the extraction properties of the carboxylic acids on the atomic number of the lanthanide shows a definite relationship to the steric bulk of the carboxylic acid molecule quantified by means of the steric parameter, E{sub s}{prime} of the substituent alkyl group. The stoichiometries of the extracted complexes for representative light (La), middle (Gd) and heavy (Lu) rare-earth metals were investigated by the slope-analysis technique for a sterically hindered acid (Versatic 10 acid; -E{prime}{sub s} = 3.83) and an acid with low steric hindrance (3-cyclohexylpropanoic acid; -E{prime}{sub s} = 0.28). 14 refs., 13 figs., 3 tabs.

  16. Extraction of uranium by macroporous bifunctional phosphinic acid resin

    SciTech Connect

    Sabharwal, K.N.; Rao, P.R.V.; Srinivasan, M.

    1995-05-01

    The extraction of U(VI), Th(IV) and a number of fission products from nitric acid medium by a newly synthesised macroporous bifunctional phosphinic acid resin has been studied. The extraction of uranium from sulphuric acid medium has also been studied. While the gel type phosphinic acid resins seems to pose a number of problems in practical applications, the macroporous type resins are shown to be suitable for a variety of applications where conventional ion exchange resins are of limited use. 12 refs., 5 figs., 3 tabs.

  17. Extraction and recovery of methylene blue from industrial wastewater using benzoic acid as an extractant.

    PubMed

    Muthuraman, G; Teng, Tjoon Tow; Leh, Cheu Peng; Norli, I

    2009-04-15

    Liquid-liquid extraction (LLE) of methylene blue (MB) from industrial wastewater using benzoic acid (extractant) in xylene has been studied at 27 degrees C. The extraction of the dye increased with increasing extractant concentration. The extraction abilities have been studied on benzoic acid concentration in the range of 0.36-5.8x10(-2) M. The distribution ratio of the dye is reasonably high (D=49.5) even in the presence of inorganic salts. Irrespective of the concentration of dye, extraction under optimal conditions was 90-99% after 15 min of phase separation. The extracted dye in the organic phase can be back extracted into sulphuric acid solution. The resultant recovered organic phase can be reused in succeeding extraction of dye with the yield ranging from 99 to 87% after 15 times reused, depending on the concentration of the initial feed solution. Experimental parameters examined were benzoic acid concentration, effect of diluent, effect of pH, effect of initial dye concentration, effect of equilibration time, various stripping agents, aqueous to organic phase ratio in extraction, organic to aqueous phase ratio in stripping and reusability of solvent.

  18. Influence of the protocol of fibroin extraction on the antibiotic activities of the constructed composites.

    PubMed

    Abdel-Fattah, Wafa I; Atwa, Nagwa; Ali, Ghareib W

    The effect of the solvents for silk fibroin (SF) extraction on its antimicrobial activity was studied. Extraction protocols were performed using LiBr (SFL) and Ajisawa's reagent (CaCl2:ethanol:H2O) (SFC). The morphological and structural characteristics of the extracted SF and their composites were assessed. Corresponding bactericidal activities against Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 25922) and Pseudomonas aeroginosa (ATCC 27853) were performed. The resulting solutions were either casted into films or individually incorporated into composites of silver nanoparticles (NS) embedded into chitosan fragments (Cs) through γ-irradiation. Films of SF, obtained by using the two solvents, as well as the final prepared composites of SF, NS and Cs were analyzed using XRD, FTIR, SEM, TEM and zeta potential at several pH values. The band gap values were calculated. The results proved that, although SFC consumed shorter gelation time, yet SFL exerted higher antibiotic activity against the tested microorganisms. Moreover, the final composites had the ability to significantly reduce the growth of these medically relevant bacteria and are, therefore, recommended as a novel natural antibacterial biomaterial for several biomedical applications.

  19. Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction.

    PubMed

    Nguyen, Quynh Thu; Wallner, Ulrike; Schmicke, Marion; Waberski, Dagmar; Henning, Heiko

    2016-11-15

    Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05), which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa.

  20. Energy metabolic state in hypothermically stored boar spermatozoa using a revised protocol for efficient ATP extraction

    PubMed Central

    Nguyen, Quynh Thu; Wallner, Ulrike; Schmicke, Marion; Waberski, Dagmar

    2016-01-01

    ABSTRACT Mammalian spermatozoa utilize ATP as the energy source for key functions on the route to fertilization. ATP and its precursor nucleotides ADP and AMP are regularly investigated in sperm physiology studies, mostly by bioluminescence assays. Assay results vary widely, mainly due to different efficiencies in nucleotide extraction and prevention of their enzymatic degradation. Here, we describe a revised, validated protocol for efficient phosphatase inhibition and adenine nucleotide extraction resulting in consistently high ATP concentrations exceeding previously reported values for boar spermatozoa up to 20-fold. The revised assay is applicable for determining ATP concentrations and adenylate energy charge in extracts from fresh and frozen samples, thereby allowing simultaneous assessment of semen samples from long-term storage experiments. After validation, the assay was applied to liquid-preserved boar spermatozoa stored at 17°C and 5°C for 24 and 72 h. Cooling to 5°C, but not storage duration, reduced ATP concentration in spermatozoa (P<0.05), which was accompanied by the appearance of AMP and ADP in the preservation medium. ATP and energy charge were highly correlated to the proportion of membrane-intact spermatozoa, supporting the idea of nucleotides leaking through disrupted membranes in cold-shocked cells. The present assay allows highly standardized studies of energy metabolism in spermatozoa. PMID:27612509

  1. A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots.

    PubMed

    Rodrigues, Elisete Pains; Torres, Adalgisa Ribeiro; da Silva Batista, Jesiane Stefânia; Huergo, Luciano; Hungria, Mariangela

    2012-06-01

    Sample preparation is a critical step in two-dimensional gel electrophoresis (2-DE) of plant tissues. Here we describe a phenol/SDS procedure that, although greatly simplified, produced well-resolved and reproducible 2-DE profiles of protein extracts from soybean [Glycine max (L.) Merril] roots. Extractions were made in three replicates using both the original and simplified procedure. To evaluate the quality of the extracted proteins, ten spots were randomly selected and identified by mass spectrometry (MS). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database entries and allowed high-quality identification of proteins. The simplified protocol described here presents considerable savings of time and reagents without compromising the quality of 2-DE protein profiles and compatibility with MS analysis, and may facilitate the progress of proteomics studies of legume-rhizobia interactions.

  2. A rapid and hazardous reagent free protocol for genomic DNA extraction suitable for genetic studies in plants.

    PubMed

    Kotchoni, Simeon O; Gachomo, Emma W

    2009-07-01

    Protocols for genomic DNA extraction from plants are generally lengthy, since they require that tissues be ground in liquid nitrogen, followed by a precipitation step, washing and drying of the DNA pellet, etc. This represents a major challenge especially when several hundred samples must be screened/analyzed within a working day. There is therefore a need for a rapid and simple procedure, which will produce DNA quality suitable for various analyses. Here, we describe a time and cost efficient protocol for genomic DNA isolation from plants suitable for all routine genetic screening/analyses. The protocol is free from hazardous reagents and therefore safe to be executed by non-specialists. With this protocol more than 100 genomic DNA samples could manually be extracted within a working day, making it a promising alternative in genetic studies of large-scale genomic screening projects.

  3. Mitotic apparatus: the selective extraction of protein with mild acid.

    PubMed

    Bibring, T; Baxandall, J

    1968-07-26

    The treatment of isolated mitotic apparatus with mild (pH 3) hydrochloric acid results in the extraction of less than 10 percent of its protein, accompanied by the selective morphological disappearance of the microtubules. The same extraction can be shown to dissolve outer doublet microtubules from sperm flagella. A protein with points of similarity to the flagellar microtubule protein is the major component of the extract from mitotic apparatus.

  4. Recovery of organic extractant from secondary emulsions formed in the extraction of uranium from wet-process phosphoric acid

    SciTech Connect

    Korchnak, J.D.; Fett, R.H.G.

    1984-01-03

    Uranium in wet-process phosphoric acid is extracted with an organic extractant. The pregnant extractant is then centrifuged to separate contaminants from the extractant. Secondary emulsions obtained by separating the contaminants following centrifugation are mixed with water or an acid leaching solution. After mixing, the mixture is centrifuged to separate and recover extractant which is recycled for stripping.

  5. Recovery of acetic acid from waste streams by extractive distillation.

    PubMed

    Demiral, H; Yildirim, M Ercengiz

    2003-01-01

    Wastes have been considered to be a serious worldwide environmental problem in recent years. Because of increasing pollution, these wastes should be treated. However, industrial wastes can contain a number of valuable organic components. Recovery of these components is important economically. Using conventional distillation techniques, the separation of acetic acid and water is both impractical and uneconomical, because it often requires large number of trays and a high reflux ratio. In practice special techniques are used depending on the concentration of acetic acid. Between 30 and 70% (w/w) acetic acid contents, extractive distillation was suggested. Extractive distillation is a multicomponent-rectification method similar in purpose to azeotropic distillation. In extractive distillation, to a binary mixture which is difficult or impossible to separate by ordinary means, a third component termed an entrainer is added which alters the relative volatility of the original constituents, thus permitting the separation. In our department acetic acid is used as a solvent during the obtaining of cobalt(III) acetate from cobalt(II) acetate by an electrochemical method. After the operation, the remaining waste contains acetic acid. In thiswork, acetic acid which has been found in this waste was recovered by extractive distillation. Adiponitrile and sulfolane were used as high boiling solvents and the effects of solvent feed rate/solution feed rate ratio and type were investigated. According to the experimental results, it was seem that the recovery of acetic acid from waste streams is possible by extractive distillation.

  6. Development of an RNA extraction protocol for detection of waterborne viruses by reverse transcriptase quantitative PCR (RT-qPCR).

    PubMed

    Jothikumar, N; Sobsey, M D; Cromeans, T L

    2010-10-01

    RNA extraction from environmental samples yields frequently an RNA preparation containing inhibitors of molecular reactions. Commercial RNA extraction kits commonly permit extraction of only 0.1-0.2 ml sample volume. An RNA extraction buffer (RNAX buffer) was formulated for the extraction of viral RNA from 4.0 ml using a silica column based protocol. To evaluate the RNAX buffer based protocol, we used hepatitis A virus (HAV) and coxsackievirus B3 (CVB3) to monitor the RNA extraction efficiency from environmental samples. For evaluation of viral RNA recovery from water concentrates which were prepared from river and pond water by PEG concentration, serial ten fold dilutions of two waterborne viruses were added to the water concentrates for evaluation by quantitative detection. Quantitative recovery of HAV and CVB3 was determined by reverse transcriptase quantitative real-time PCR (RT-qPCR). The extracted RNA was compatible with RT-qPCR and sensitivity of detection of 0.8PFU per reaction was found with RNAX buffer and the developed protocol. This level of sensitivity was obtained using viral RNA extracted from 4.0 ml of an inoculated water sample concentrate. The RNAX buffer developed in this study could be applicable to the detection of other pathogens in water and food.

  7. Comparison of five protocols to extract DNA from paraffin-embedded tissues for the detection of human papillomavirus.

    PubMed

    Alvarez-Aldana, Adalucy; Martínez, José William; Sepúlveda-Arias, Juan C

    2015-02-01

    Formalin-fixed paraffin-embedded (FFPE) tissues are a valuable source of DNA with which to perform large retrospective studies on the epidemiology of HPV infection. Five different DNA extraction protocols were carried out to evaluate the DNA obtained from FFPE samples with polymerase chain reaction (PCR) using two primer sets to amplify a constitutive human gene, β-globin, and two primer sets to detect the L1 and E6 HPV genes. From the five DNA extraction protocols evaluated, the best results were obtained with protocol A, corresponding to a crude extract from the sample. With the procedures described herein, we were able to amplify DNA extracted from archival paraffin blocks stored for six years. However, the amplification products were more efficiently obtained with primers that amplified shorter fragments. This result indicates that a major factor limiting the extraction process in these samples is DNA fragmentation, a factor that will naturally vary between the different specimens evaluated. Also, depending upon the extraction method, PCR amplification of a human gene does not necessarily guarantee the successful extraction of viral DNA. In conclusion, different DNA and HPV detection methods can significantly influence the results. Therefore, the DNA extraction methods and primers used for DNA amplification in fixed tissues need to be chosen carefully, depending on the specific requirements of the study being carried out.

  8. Electromembrane extraction and HPLC analysis of haloacetic acids and aromatic acetic acids in wastewater.

    PubMed

    Alhooshani, Khalid; Basheer, Chanbasha; Kaur, Jagjit; Gjelstad, Astrid; Rasmussen, Knut E; Pedersen-Bjergaard, Stig; Lee, Hian Kee

    2011-10-30

    For the first time, haloacetic acids and aromatic acetic acids were extracted from wastewater samples using electromembrane extraction (EME). A thin layer of toluene immobilized on the walls of a polypropylene membrane envelope served as an artificial supported liquid membrane (SLM). The haloacetic acids (HAAs) (chloroacetic acid, dichloroacetic acid, and trifluoroacetic acid) and aromatic acetic acids (phenylacetic acid and p-hydroxyphenylacetic acid) were extracted through the SLM and into an alkalized aqueous buffer solution. The buffer solution was located inside the membrane envelope. The electrical potential difference sustained over the membrane acted as the driving force for the transport of haloacetic acids into the membrane by electrokinetic migration. After extraction, the extracts were analyzed by high-performance liquid chromatography-ultraviolet detection. The detection limits were between 0.072 and 40.3 ng L(-1). The calibration plot linearity was in the range of 5 and 200 μg L(-1) while the correlation coefficients for the analytes ranged from 0.9932 to 0.9967. Relative recoveries were in the range of 87-106%. The extraction efficiency was found to be comparable to that of solid-phase extraction.

  9. A simple protocol for protein extraction of recalcitrant fruit tissues suitable for 2-DE and MS analysis.

    PubMed

    Song, Jun; Braun, Gordon; Bevis, Eric; Doncaster, Kristen

    2006-08-01

    Fruit tissues are considered recalcitrant plant tissue for proteomic analysis. Three phenol-free protein extraction procedures for 2-DE were compared and evaluated on apple fruit proteins. Incorporation of hot SDS buffer, extraction with TCA/acetone precipitation was found to be the most effective protocol. The results from SDS-PAGE and 2-DE analysis showed high quality proteins. More than 500 apple polypeptides were separated on a small scale 2-DE gel. The successful protocol was further tested on banana fruit, in which 504 and 386 proteins were detected in peel and flesh tissues, respectively. To demonstrate the quality of the extracted proteins, several protein spots from apple and banana peels were cut from 2-DE gels, analyzed by MS and have been tentatively identified. The protocol described in this study is a simple procedure which could be routinely used in proteomic studies of many types of recalcitrant fruit tissues.

  10. Extraction of protactinium from mineral acid-alcohol media.

    PubMed

    Alian, A; Sanad, W; Shabana, R

    1968-07-01

    The extraction of protactinium with organic solvents has been investigated in the presence of water-miscible alcohols and acetone. These additives were found to increase considerably the extraction of protactinium in the cases of trilaurylamine, tributyl phosphate and isobutyl methyl ketone. The influence was less in the case of thenoyltrifluoroacetone. In mixtures of an acid with various alcohols, the influence depended on the alcohol concentration, the acidity and on the chain lengths and dielectric constants of the alcohol introduced into the extraction system.

  11. Approaches for regeneration of amine-carboxylic acid extracts

    SciTech Connect

    Dai, Y.; King, C.J.

    1995-07-01

    Extraction processes based on reversible chemical complexation can be useful for separation of polar organics from dilute solution. Tertiary amines are effective extractants for the recovery of carboxylic acids from aqueous solution. The regeneration of aminecarboxylic acid extracts is an important step which strongly influences the economic viability of the separation process. Several regeneration methods are critically reviewed, and the factors that affect swing regeneration processes, including temperature-swing, diluent composition-swing and pH-swing with a volatile base are discussed. Interest in this area comes from interest in treatment of waste streams, particularly in petrochemical and fermentation manufacture.

  12. Transparent DNA/RNA Co-extraction Workflow Protocol Suitable for Inhibitor-Rich Environmental Samples That Focuses on Complete DNA Removal for Transcriptomic Analyses.

    PubMed

    Lim, Natalie Y N; Roco, Constance A; Frostegård, Åsa

    2016-01-01

    Adequate comparisons of DNA and cDNA libraries from complex environments require methods for co-extraction of DNA and RNA due to the inherent heterogeneity of such samples, or risk bias caused by variations in lysis and extraction efficiencies. Still, there are few methods and kits allowing simultaneous extraction of DNA and RNA from the same sample, and the existing ones generally require optimization. The proprietary nature of kit components, however, makes modifications of individual steps in the manufacturer's recommended procedure difficult. Surprisingly, enzymatic treatments are often performed before purification procedures are complete, which we have identified here as a major problem when seeking efficient genomic DNA removal from RNA extracts. Here, we tested several DNA/RNA co-extraction commercial kits on inhibitor-rich soils, and compared them to a commonly used phenol-chloroform co-extraction method. Since none of the kits/methods co-extracted high-quality nucleic acid material, we optimized the extraction workflow by introducing small but important improvements. In particular, we illustrate the need for extensive purification prior to all enzymatic procedures, with special focus on the DNase digestion step in RNA extraction. These adjustments led to the removal of enzymatic inhibition in RNA extracts and made it possible to reduce genomic DNA to below detectable levels as determined by quantitative PCR. Notably, we confirmed that DNase digestion may not be uniform in replicate extraction reactions, thus the analysis of "representative samples" is insufficient. The modular nature of our workflow protocol allows optimization of individual steps. It also increases focus on additional purification procedures prior to enzymatic processes, in particular DNases, yielding genomic DNA-free RNA extracts suitable for metatranscriptomic analysis.

  13. Transparent DNA/RNA Co-extraction Workflow Protocol Suitable for Inhibitor-Rich Environmental Samples That Focuses on Complete DNA Removal for Transcriptomic Analyses

    PubMed Central

    Lim, Natalie Y. N.; Roco, Constance A.; Frostegård, Åsa

    2016-01-01

    Adequate comparisons of DNA and cDNA libraries from complex environments require methods for co-extraction of DNA and RNA due to the inherent heterogeneity of such samples, or risk bias caused by variations in lysis and extraction efficiencies. Still, there are few methods and kits allowing simultaneous extraction of DNA and RNA from the same sample, and the existing ones generally require optimization. The proprietary nature of kit components, however, makes modifications of individual steps in the manufacturer’s recommended procedure difficult. Surprisingly, enzymatic treatments are often performed before purification procedures are complete, which we have identified here as a major problem when seeking efficient genomic DNA removal from RNA extracts. Here, we tested several DNA/RNA co-extraction commercial kits on inhibitor-rich soils, and compared them to a commonly used phenol-chloroform co-extraction method. Since none of the kits/methods co-extracted high-quality nucleic acid material, we optimized the extraction workflow by introducing small but important improvements. In particular, we illustrate the need for extensive purification prior to all enzymatic procedures, with special focus on the DNase digestion step in RNA extraction. These adjustments led to the removal of enzymatic inhibition in RNA extracts and made it possible to reduce genomic DNA to below detectable levels as determined by quantitative PCR. Notably, we confirmed that DNase digestion may not be uniform in replicate extraction reactions, thus the analysis of “representative samples” is insufficient. The modular nature of our workflow protocol allows optimization of individual steps. It also increases focus on additional purification procedures prior to enzymatic processes, in particular DNases, yielding genomic DNA-free RNA extracts suitable for metatranscriptomic analysis. PMID:27803690

  14. Nucleic acids extraction from laser microdissected FFPE tissue sections.

    PubMed

    Burgemeister, Renate

    2011-01-01

    Tissue heterogeneity is a common source of unsuccessful experiments. Laser capture microdissection is a tool to prepare homogeneous tissue and cell areas as starting material for reliable and reproducible results as it allows the defined investigation of spatially different tissue areas.Nearly all samples allow the extraction of DNA. Fresh or fresh frozen samples are an ideal source for getting access to high-quality RNA. But also the large archives of formalin-fixed, paraffin-embedded (FFPE) tissue specimens are a valuable source of sample material for RNA extraction. Optimized protocols may help to make the RNA from FFPE material suitable for expression studies.

  15. Comprehensive extraction study using N,N-dioctyldiglycolamic acid.

    PubMed

    Shimojo, Kojiro; Nakai, Ayaka; Okamura, Hiroyuki; Saito, Takumi; Ohashi, Akira; Naganawa, Hirochika

    2014-01-01

    We report on the acid dissociation constants (Ka) of diglycolamic acid-type ligands together with comprehensive data on the extraction performance of N,N-dioctyldiglycolamic acid (DODGAA) for 54 metal ions. The pKa of the diglycolamic acid framework was determined to be 3.54 ± 0.03 in water (0.1 M LiCl, 25°C) by potentiometric titration, indicating that DODGAA is strongly acidic compared with acetic acid. DODGAA can quantitatively transfer various metal ions among the 54 metal ions through a proton-exchange reaction, and provides excellent extraction performance and separation ability for rare-earth metal ions, In(III), Fe(III), Hg(II), and Pb(II) among the 54 metal ions.

  16. A “Novel” Protocol for the Analysis of Hydroxycinnamic Acids in Leaf Tissue of Chicory (Cichorium intybus L., Asteraceae)

    PubMed Central

    Bahri, Meriem; Hance, Philippe; Grec, Sébastien; Quillet, Marie-Christine; Trotin, Francis; Hilbert, Jean-Louis; Hendriks, Theo

    2012-01-01

    A “novel” protocol is presented for easy and reliable estimation of soluble hydroxycinnamate levels in Cichorium intybus L. leaf tissue in large-scale experiments. Samples were standardized by punching 6 discs per leaf, and hydroxycinnamates were extracted by submerging the discs in 80% ethanol with 5% acetic acid for at least 48 h in the darkness at 4°C. Residual dry mass of the discs was used for a posteriori correction of compound levels. Chlorophyll was eliminated by chloroform, and the aqueous phases were transferred to microplates, dried, and dissolved in 50% methanol for HPLC analysis and storage. An HPLC program of 8 min was developed for the analysis of the extracts. Comparisons with extractions of liquid nitrogen powders indicated that the novel extraction method was reliable. No degradation of the major hydroxycinnamates—caftaric, chlorogenic, and chicoric acids—was observed, during maceration at ambient temperatures, or after storage for 1 year. PMID:23304076

  17. Fatty and resinic acids extractions from crude tall oil

    SciTech Connect

    Nogueira, J.M.F.

    1996-11-01

    The separation of fatty and resinic acidic fractions from crude tall-oil soap solutions with n-heptane by the technique of dissociation extraction is discussed. The theory of the overall process is supported by a systematic study developed to cover the high selectivity demonstrated in the differential solubility and the aptness between fatty and diterpenic acids to both liquids phases. To study the main factors affecting those liquid-liquid extraction systems and the amphiphilic behavior of such molecules involved, sodium salts aqueous solutions of crude tall oil and synthetic mixtures as molecular acidic models were used.

  18. DNA extractions from deep subseafloor sediments: novel cryogenic-mill-based procedure and comparison to existing protocols.

    PubMed

    Alain, Karine; Callac, Nolwenn; Ciobanu, Maria-Cristina; Reynaud, Yann; Duthoit, Frédérique; Jebbar, Mohamed

    2011-12-01

    Extracting DNA from deep subsurface sediments is challenging given the complexity of sediments types, low biomasses, resting structures (spores, cysts) frequently encountered in deep sediments, and the potential presence of enzymatic inhibitors. Promising results for cell lysis efficiency were recently obtained by use of a cryogenic mill (Lipp et al., 2008). These findings encouraged us to devise a DNA extraction protocol using this tool. Thirteen procedures involving a combination of grinding in liquid nitrogen (for various durations and beating rates) with different chemical solutions (phenol, chloroform, SDS, sarkosyl, proteinase, GTC), or with use of DNA recovery kits (MagExtractor®) were compared. Effective DNA extraction was evaluated in terms of cell lysis efficiency, DNA extraction efficiency, DNA yield and determination of prokaryotic diversity. Results were compared to those obtained by standard protocols: the FastDNA®SPIN kit for soil and the Zhou protocol. For most sediment types grinding in a cryogenic mill at a low beating rate in combination with direct phenol-chloroform extraction resulted in much higher DNA yields than those obtained using classical procedures. In general (except for clay-rich sediments), this procedure provided high-quality crude extracts for direct downstream nested-PCR, from cell numbers as low as 1.1×10(6) cells/cm(3). This procedure is simple, rapid, low-cost, and could be used with minor modifications for large-scale DNA extractions for a variety of experimental goals.

  19. Development of an eco-protocol for seaweed chlorophylls extraction and possible applications in dye sensitized solar cells

    NASA Astrophysics Data System (ADS)

    Armeli Minicante, S.; Ambrosi, E.; Back, M.; Barichello, J.; Cattaruzza, E.; Gonella, F.; Scantamburlo, E.; Trave, E.

    2016-07-01

    Seaweeds are a reserve of natural dyes (chlorophylls a, b and c), characterized by low cost and easy supply, without potential environmental load in terms of land subtraction, and also complying with the requirements of an efficient waste management policy. In particular, the brown seaweed Undaria pinnatifida is a species largely present in the Venice Lagoon area, and for it a removal strategy is actually mandatory. In this paper, we set-up an eco-protocol for the best extraction and preparation procedures of the pigment, with the aim of finding an easy and affordable method for chlorophyll c extraction, exploring at the same time the possibility of using these algae within local sustainable management integrated strategies, among which the possible use of chlorophylls as a dye source in dye sensitized solar cells (DSSCs) is investigated. Experimental results suggest that the developed protocols are useful to optimize the chlorophyll c extraction, as shown by optical absorption spectroscopy measurements. The DSSCs built with the chlorophyll extracted by the proposed eco-protocol exhibit solar energy conversion efficiencies are similar to those obtained following extraction protocols with larger environmental impacts.

  20. A simplified genomic DNA extraction protocol for pre-germination genotyping in rice.

    PubMed

    Duan, Y B; Zhao, F L; Chen, H D; Li, H; Ni, D H; Wei, P C; Sheng, W; Teng, J T; Zhang, A M; Xue, J P

    2015-06-11

    Genotyping is a critical step for molecular marker-assisted selection in rice. Rice genomic DNA samples for genotyping are typically isolated from living tissues such as seedlings. This requires the germination of all candidate seeds and extraction of DNA from the seedlings. Currently, an ideal individual is selected from a very large number of plants, which is time- and labor-consuming, requiring several transplantations of materials and sampling processes. In this study, we developed a simplified genomic DNA extraction protocol in rice by using amylase to treat half-seeds. The yields of genomic DNA from a half-seed of Indica and Japonica rice were greater than 203.8 ± 32.5 and 143.2 ± 25.5 ng, respectively, and the 260/280 nm absorbance ratio was 1.75-2.10. The DNA was confirmed to be sufficient for polymerase chain reaction amplification and can be used in a marker-assisted selection program.

  1. Microwave-assisted extraction of glycyrrhizic acid from licorice root.

    PubMed

    Pan; Liu; Jia; Shu

    2000-07-01

    In the present study, a microwave-assisted extraction (MAE) technique has been developed for the extraction of glycyrrhizic acid (GA) from licorice root. Various experimental conditions, such as extraction time, different ethanol and ammonia concentration, liquid/solid ratios, pre-leaching time before MAE and material size for the MAE procedure were investigated to optimize the efficiency of the extraction. Under appropriate MAE conditions, such as extraction times of 4-5min, ethanol concentrations of 50-60% (v/v), ammonia concentrations of 1-2% (v/v) and liquid/solid ratios of 10:1(ml/g), the recovery of GA from licorice root with MAE was equivalent with conventional extraction methods. Those methods include extraction at room temperature (ERT), the traditional Soxhlet extraction, heat reflux extraction and ultrasonic extraction. Due to the considerable savings in time and solvent, MAE was more effective than the conventional methods. This novel method is suitable for fast extraction of GA from licorice root.

  2. Supercritical fluid extraction of free amino acids from broccoli leaves.

    PubMed

    Arnáiz, E; Bernal, J; Martín, M T; Nozal, M J; Bernal, J L; Toribio, L

    2012-08-10

    The extraction of free amino acids (AAs) from broccoli leaves using supercritical fluid extraction (SFE) with CO(2) modified with methanol, is presented in this work. The effect of the different variables was studied, showing the percentage of methanol a strong influence on the extraction. The best results in terms of extraction yield were obtained at 250 bar, 70°C, 35% methanol as organic modifier, a flow rate of 2 mL/min, and 5 min and 30 min as static and dynamic extraction times, respectively. The extraction yield obtained with the SFE method was comparable to that obtained employing conventional solvent extraction with methanol-water (70:30) and minor than using water, but the relative proportion of the AAs in the extracts was very different. For example, the use of SFE allowed the enrichment in proline and glutamine of the extracts. The selected conditions were applied to obtain SFE extracts of broccoli leaves from different varieties (Naxos, Nubia, Marathon, Parthenon and Viola). The highest levels of AAs were found in the SFE extracts from the Nubia variety.

  3. A 1-dodecanethiol-based phase transfer protocol for the highly efficient extraction of noble metal ions from aqueous phase.

    PubMed

    Chen, Dong; Cui, Penglei; Cao, Hongbin; Yang, Jun

    2015-03-01

    A 1-dodecanethiol-based phase-transfer protocol is developed for the extraction of noble metal ions from aqueous solution to a hydrocarbon phase, which calls for first mixing the aqueous metal ion solution with an ethanolic solution of 1-dodecanethiol, and then extracting the coordination compounds formed between noble metal ions and 1-dodecanethiol into a non-polar organic solvent. A number of characterization techniques, including inductively coupled plasma atomic emission spectroscopy, Fourier transform infrared spectroscopy, and thermogravimetric analysis demonstrate that this protocol could be applied to extract a wide variety of noble metal ions from water to dichloromethane with an efficiency of >96%, and has high selectivity for the separation of the noble metal ions from other transition metals. It is therefore an attractive alternative for the extraction of noble metals from water, soil, or waste printed circuit boards.

  4. Enhanced lipid extraction from algae using free nitrous acid pretreatment.

    PubMed

    Bai, Xue; Naghdi, Forough Ghasemi; Ye, Liu; Lant, Paul; Pratt, Steven

    2014-05-01

    Lipid extraction has been identified as a major bottleneck for large-scale algal biodiesel production. In this work free nitrous acid (FNA) is presented as an effective and low cost pretreatment to enhance lipid recovery from algae. Two batch tests, with a range of FNA additions, were conducted to disrupt algal cells prior to lipid extraction by organic solvents. Total accessible lipid content was quantified by the Bligh and Dyer method, and was found to increase with pretreatment time (up to 48 h) and FNA concentration (up to 2.19 mg HNO2-N/L). Hexane extraction was used to study industrially accessible lipids. The mass transfer coefficient (k) for lipid extraction using hexane from algae treated with 2.19 mg HNO2-N/L FNA was found to be dramatically higher than for extraction from untreated algae. Consistent with extraction results, cell disruption analysis indicated the disruption of the cell membrane barrier.

  5. New protein extraction/solubilization protocol for gel-based proteomics of rat (female) whole brain and brain regions.

    PubMed

    Hirano, Misato; Rakwal, Randeep; Shibato, Junko; Agrawal, Ganesh Kumar; Jwa, Nam-Soo; Iwahashi, Hitoshi; Masuo, Yoshinori

    2006-08-31

    The rat is an accepted model for studying human psychiatric/neurological disorders. We provide a protocol for total soluble protein extraction using trichloroacetic acid/acetone (TCA/A) from rat (female) whole brain, 10 brain regions and the pituitary gland, and show that two-dimensional gel electrophoresis (2-DGE) using pre-cast immobilized pH (4-7) gradient (IPG) strip gels (13 cm) in the first dimension yields clean silver nitrate stained protein profiles. Though TCA/A precipitation may not be "ideal", the important choice here is the selection of an appropriate lysis buffer (LB) for solubilizing precipitated proteins. Our results reveal enrichment of protein spots by use of individual brain regions rather than whole brain, as well as the presence of differentially expressed spots in their proteomes. Thus individual brain regions provide improved protein coverage and are better suited for differential protein detection. Moreover, using a phosphoprotein-specific dye, in-gel detection of phosphoproteins was demonstrated. Representative high-resolution silver nitrate stained proteome profiles of rat whole brain total soluble protein are presented. Shortcomings apart (failure to separate membrane proteins), gel-based proteomics remains a viable option, and 2-DGE is the method of choice for generating high-resolution proteome maps of rat brain and brain regions.

  6. Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis.

    PubMed

    Zakharchenko, Olena; Greenwood, Christina; Alldridge, Louise; Souchelnytskyi, Serhiy

    2011-03-10

    Proteomics is a highly informative approach to analyze cancer-associated transformation in tissues. The main challenge to use a tissue for proteomics studies is the small sample size and difficulties to extract and preserve proteins. The choice of a buffer compatible with proteomics applications is also a challenge. Here we describe a protocol optimized for the most efficient extraction of proteins from the human breast tissue in a buffer compatible with two-dimensional gel electrophoresis (2D-GE). This protocol is based on mechanically assisted disintegration of tissues directly in the 2D-GE buffer. Our method is simple, robust and easy to apply in clinical practice. We demonstrate high quality of separation of proteins prepared according to the reported here protocol.

  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. The extraction of actinides from nitric acid solutions with diamides of dipicolinic acid

    NASA Astrophysics Data System (ADS)

    Lapka, Joseph L.; Paulenova, Alena; Alyapyshev, Mikhail Yu; Babain, Vasiliy A.; Law, Jack D.; Herbst, R. Scott

    2010-03-01

    Diamides of dipicolinic acid (N,N'-diethyl-N,N'-ditolyl-dipicolinamide, EtTDPA) were synthesized and evaluated for their extraction capability for actinides. In this work the extractions of neptunium(V), protactinium(V), and thorium(IV) with EtTDPA in a polar fluorinated diluent from nitric acid were investigated. EtTDPA shows a high affinity for Th(IV) even at millimolar concentrations. Np(V) and Pa(V) are both reasonably extractable with EtTDPA; however, near saturated solutions are required to achieve appreciable distribution ratios. A comparison with previously published actinide extraction data is given.

  9. Separation of Minor Actinides from Lanthanides by Dithiophosphinic Acid Extractants

    SciTech Connect

    D. R. Peterman; M. R. Greenhalgh; R. D. Tillotson; J. R. Klaehn; M. K. Harrup; T. A. Luther; J. D. Law; L. M. Daniels

    2008-09-01

    The selective extraction of the minor actinides (Am(III) and Cm(III)) from the lanthanides is an important part of advanced reprocessing of spent nuclear fuel. This separation would allow the Am/Cm to be fabricated into targets and recycled to a reactor and the lanthanides to be dispositioned. This separation is difficult to accomplish due to the similarities in the chemical properties of the trivalent actinides and lanthanides. Research efforts at the Idaho National Laboratory have identified an innovative synthetic pathway yielding new regiospecific dithiophosphinic acid (DPAH) extractants. The synthesis provides DPAH derivatives that can address the issues concerning minor actinide separation and extractant stability. For this work, two new symmetric DPAH extractants have been prepared. The use of these extractants for the separation of minor actinides from lanthanides will be discussed.

  10. DNA Extraction Protocol for Plants with High Levels of Secondary Metabolites and Polysaccharides without Using Liquid Nitrogen and Phenol.

    PubMed

    Sahu, Sunil Kumar; Thangaraj, Muthusamy; Kathiresan, Kandasamy

    2012-01-01

    Mangroves and salt marsh species are known to synthesize a wide spectrum of polysaccharides and polyphenols including flavonoids and other secondary metabolites which interfere with the extraction of pure genomic DNA. Although a plethora of plant DNA isolation protocols exist, extracting DNA from mangroves and salt marsh species is a challenging task. This study describes a rapid and reliable cetyl trimethylammonium bromide (CTAB) protocol suited specifically for extracting DNA from plants which are rich in polysaccharides and secondary metabolites, and the protocol also excludes the use of expensive liquid nitrogen and toxic phenols. Purity of extracted DNA was excellent as evident by A260/A280 ratio ranging from 1.78 to 1.84 and A260/A230 ratio was >2, which also suggested that the preparations were sufficiently free of proteins and polyphenolics/polysaccharide compounds. DNA concentration ranged from 8.8 to 9.9 μg μL(-1). The extracted DNA was amenable to RAPD, restriction digestion, and PCR amplification of plant barcode genes (matK and rbcl). The optimized method is suitable for both dry and fresh leaves. The success of this method in obtaining high-quality genomic DNA demonstrated the broad applicability of this method.

  11. A comparative study of three extraction protocols of DNA from nails: Potential use in the diagnosis of onychomycoses.

    PubMed

    Kotrotsiou, T; Chatzimichaloglou, A; Nasr, A; Spandou, E; Patsatsi, A; Sotiriadis, D; Vyzantiadis, T-A

    2017-03-01

    Molecular techniques can be useful adjuncts to the diagnosis of onychomycoses. However, the nail presents difficulties in the extraction of its DNA. The comparison of three extraction protocols of DNA from nails and their ranking for possible use in the molecular diagnosis of onychomycoses are described. Extraction was performed on weighed nail clippings of equal size from positive (31) or negative (14) samples, according to the culture result. At Prot1, the extraction was performed according to Tahir and Watson, with an additional step implementing silica columns. At Prot2, the methodology proposed by the Statens Serum Institute of Copenhagen was used. At Prot3, DNA was extracted by the use of magnetic separation after homogenisation with glass beading. The evaluation parameters were DNA purity, DNA concentration, total DNA yield/g of tissue, cost and duration. The multiples of the means of medians of the first three parameters, for each protocol, were calculated. Prot3 showed the highest DNA purity. Prot2 presented the highest DNA concentration and DNA yield/g of tissue, while it was the cheapest and shortest. In total, the three protocols were graded as Prot2>Prot1>Prot3. The second method, although had a lower DNA purity, presented the higher DNA concentration and DNA yield, while its duration and cost were also favourable.

  12. Rosmarinic Acid and Melissa officinalis Extracts Differently Affect Glioblastoma Cells.

    PubMed

    Ramanauskiene, Kristina; Raudonis, Raimondas; Majiene, Daiva

    Lemon balm (Melissa officinalis L.) has many biological effects but especially important is its neuroprotective activity. The aim of the study is to produce different extracts of Melissa officinalis and analyse their chemical composition and biological properties on rat glioblastoma C6 cells. Results revealed that rosmarinic acid (RA) is the predominant compound of lemon balm extracts. RA has cytotoxic effect on glioblastoma cells (LC50 290.5 μM after the incubation of 24 h and LC50 171.3 μM after 48 h). RA at concentration 80-130 μM suppresses the cell proliferation and has an antioxidant effect. 200 μM and higher concentrations of RA have a prooxidant effect and initiate cell death through necrosis. The aqueous extract of lemon balm is also enriched in phenolic compounds: protocatechuic, caftaric, caffeic, ferulic, and cichoric acids and flavonoid luteolin-7-glucoside. This extract at concentrations 50 μM-200 μM RA has cytotoxic activity and initiates cell death through apoptosis. Extracts prepared with 70% ethanol contain the biggest amount of active compounds. These extracts have the highest cytotoxic activity on glioblastoma cells. They initiate generation of intracellular ROS and cell death through apoptosis and necrosis. Our data suggest that differently prepared lemon balm extracts differently affect glioblastoma cells and can be used as neuroprotective agents in several therapeutic strategies.

  13. Rosmarinic Acid and Melissa officinalis Extracts Differently Affect Glioblastoma Cells

    PubMed Central

    Ramanauskiene, Kristina; Raudonis, Raimondas

    2016-01-01

    Lemon balm (Melissa officinalis L.) has many biological effects but especially important is its neuroprotective activity. The aim of the study is to produce different extracts of Melissa officinalis and analyse their chemical composition and biological properties on rat glioblastoma C6 cells. Results revealed that rosmarinic acid (RA) is the predominant compound of lemon balm extracts. RA has cytotoxic effect on glioblastoma cells (LC50 290.5 μM after the incubation of 24 h and LC50 171.3 μM after 48 h). RA at concentration 80–130 μM suppresses the cell proliferation and has an antioxidant effect. 200 μM and higher concentrations of RA have a prooxidant effect and initiate cell death through necrosis. The aqueous extract of lemon balm is also enriched in phenolic compounds: protocatechuic, caftaric, caffeic, ferulic, and cichoric acids and flavonoid luteolin-7-glucoside. This extract at concentrations 50 μM–200 μM RA has cytotoxic activity and initiates cell death through apoptosis. Extracts prepared with 70% ethanol contain the biggest amount of active compounds. These extracts have the highest cytotoxic activity on glioblastoma cells. They initiate generation of intracellular ROS and cell death through apoptosis and necrosis. Our data suggest that differently prepared lemon balm extracts differently affect glioblastoma cells and can be used as neuroprotective agents in several therapeutic strategies. PMID:27688825

  14. SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol.

    PubMed

    Paul, Tanima; Basu, Semanti; Sarkar, Keka

    2014-12-01

    Direct isolation of soil DNA comes as an emerging technology to understand the microbial diversity of a particular environment circumventing the dependency on culturable methods. Soil DNA isolation is tough due to the presence of various organic components present in soil which interfere in extraction procedure. Here, we report a novel direct soil DNA extraction protocol utilizing bare superparamagnetic iron oxide nanoparticles and its comparison with conventional and commercial kit-based soil DNA extraction methods. The quality, quantity and feasibility of the recovered DNA from all the three methods towards various molecular techniques were checked. Our magnetic nanoparticle-based soil DNA extraction successfully yields pure DNA without any RNA or protein contamination as revealed by the nanodrop spectrophotometer and agarose gel electrophoretic study. Different methods of soil DNA extraction were evaluated on the basis of PCR, denaturing gradient gel electrophoresis and real-time PCR. Soil DNA extracted using conventional method fails to carry out critical molecular biology techniques where as magnetic nanoparticle-based soil DNA extraction gave good results which is comparable to commercial kit. This comparative study suggests that protocol described in this report is novel, less time consuming, cost effective with fewer handling steps and yields high quantity, good quality DNA from soil.

  15. Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae.

    PubMed

    Mogni, Virginia Y; Kahan, Mariano A; de Queiroz, Luciano Paganucci; Vesprini, José L; Ortiz, Juan Pablo A; Prado, Darién E

    2016-01-01

    The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites-i.e. tannins and oleoresins-that are present in almost all tissues of the many members of the group, which complicate the purification and amplification of the DNA. The objective of this work was to improve an available DNA isolation method for Schinopsis spp. and other related Anacardiaceae, as well as the PCR protocols for DNA amplification of the chloroplast trnL-F, rps16 and ndhF and nuclear ITS-ETS fragments. The modifications proposed allowed the extraction of 70-120 µg of non-degraded genomic DNA per gram of dry tissue that resulted useful for PCR amplification. PCR reactions produced the expected fragments that could be directly sequenced. Sequence analyses of amplicons showed similarity with the corresponding Schinopsis accessions available at GenBank. The methodology presented here can be routinely applied for molecular studies of the group aimed to clarify not only aspects on the molecular biology but also the taxonomy and phylogeny of this fascinating group of vascular plants.

  16. Blood cell mRNAs and microRNAs: optimized protocols for extraction and preservation.

    PubMed

    Eikmans, Michael; Rekers, Niels V; Anholts, Jacqueline D H; Heidt, Sebastiaan; Claas, Frans H J

    2013-03-14

    Assessing messenger RNA (mRNA) and microRNA levels in peripheral blood cells may complement conventional parameters in clinical practice. Working with small, precious samples requires optimal RNA yields and minimal RNA degradation. Several procedures for RNA extraction and complementary DNA (cDNA) synthesis were compared for their efficiency. The effect on RNA quality of freeze-thawing peripheral blood cells and storage in preserving reagents was investigated. In terms of RNA yield and convenience, quality quantitative polymerase chain reaction signals per nanogram of total RNA and using NucleoSpin and mirVana columns is preferable. The SuperScript III protocol results in the highest cDNA yields. During conventional procedures of storing peripheral blood cells at -180°C and thawing them thereafter, RNA integrity is maintained. TRIzol preserves RNA in cells stored at -20°C. Detection of mRNA levels significantly decreases in degraded RNA samples, whereas microRNA molecules remain relatively stable. When standardized to reference targets, mRNA transcripts and microRNAs can be reliably quantified in moderately degraded (quality index 4-7) and severely degraded (quality index <4) RNA samples, respectively. We describe a strategy for obtaining high-quality and quantity RNA from fresh and stored cells from blood. The results serve as a guideline for sensitive mRNA and microRNA expression assessment in clinical material.

  17. Development of a direct in-matrix extraction (DIME) protocol for MALDI-TOF-MS detection of glycated phospholipids in heat-treated food samples.

    PubMed

    Calvano, Cosima D; De Ceglie, Cristina; Zambonin, Carlo G

    2014-09-01

    In foodstuffs, one of the main factors inducing modifications in phospholipids (PLs) structure is the heat treatment. Among PLs, only phosphatidylethanolamines and phosphatidylserines, due to their free amino group, can be involved in Maillard reaction and can form adducts with reducing sugars, besides other by-products called advanced glycation end-products. To date, glycated lipid products are less characterized in comparison to proteins. The aim of this work was to develop a novel, rapid and sensitive extraction protocol for the detection and characterization of modified PLs (glycated and oxidized) by means of matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). At first, to investigate the formation of glycated and/or short chain by-products in different classes of PLs, representative standards were heated with or without sugar (lactose or glucose) and subjected to traditional lipid extraction methods as Bligh and Dyer and to the novel direct in matrix extraction (DIME) using 1,8-bis(dimethylamino)naphthalene as preconcentrating matrix. MALDI-MS analysis in negative ion mode allowed detecting glycation and oxidation products both on fatty acid and glucose moieties. Then, the procedure was successfully applied to different heat-treated and powdered samples (milk powders, pasteurized milk, ultra-high-temperature milk and soy flour) for the detection of modified PLs in complex foods. The currently developed DIME protocol could be a powerful tool for understanding lipid glycation also in biological samples.

  18. Urinary extracellular vesicles for RNA extraction: optimization of a protocol devoid of prokaryote contamination

    PubMed Central

    Tataruch-Weinert, Dorota; Musante, Luca; Kretz, Oliver; Holthofer, Harry

    2016-01-01

    Background Urinary extracellular vesicles (UEVs) represent an ideal platform for biomarker discovery. They carry different types of RNA species, and reported profile discrepancies related to the presence/absence of 18s and 28s rRNA remain controversial. Moreover, sufficient urinary RNA yields and respective quality RNA profiles are still to be fully established. Methods UEVs were enriched by hydrostatic filtration dialysis, and RNA content was extracted using 7 different commercially available techniques. RNA quantity was assessed using spectrophotometry and fluorometry, whilst RNA quality was determined by capillary electrophoresis. Results The presence of prokaryotic transcriptome was stressed when cellular RNA, as a control, was spiked into the UEVs samples before RNA extraction. The presence of bacteria in hydrostatic filtration dialysis above 1,000 kDa molecular weight cut-off and in crude urine was confirmed with growth media plates. The efficiency in removing urinary bacteria was evaluated by differential centrifugation, filtration (0.22 µm filters) and chemical pretreatment (water purification tablet). For volumes of urine >200 ml, the chemical treatment provides ease of handling without affecting vesicle integrity, protein and RNA profiles. This protocol was selected to enrich RNA with 7 methods, and its respective quality and quantity were assessed. The results were given as follows: (a) Fluorometry gave more repeatability and reproducibility than spectrophotometry to assess the RNA yields, (b) UEVs were enriched with small RNA, (c) Ribosomal RNA peaks were not observed for any RNA extraction method used and (d) RNA yield was higher for column-based method designed for urinary exosome, whilst the highest relative microRNA presence was obtained using TRIzol method. Conclusion Our results show that the presence of bacteria can lead to misidentification in the electrophoresis peaks. Fluorometry is more reliable than spectrophotometry. RNA isolation method

  19. Improved method for extracting lanthanides and actinides from acid solutions

    DOEpatents

    Horwitz, E.P.; Kalina, D.G.; Kaplan, L.; Mason, G.W.

    1983-07-26

    A process for the recovery of actinide and lanthanide values from aqueous acidic solutions uses a new series of neutral bi-functional extractants, the alkyl(phenyl)-N,N-dialkylcarbamoylmethylphosphine oxides. The process is suitable for the separation of actinide and lanthanide values from fission product values found together in high-level nuclear reprocessing waste solutions.

  20. Acidic solvent extraction of gossypol from cottonseed meal

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In order to expand the use of cottonseed meal in animal feeding, extraction of the meal gossypol was studied with acetic acetone- and ethanol-based solutions. Phosphoric acid was added to hydrolyze and release gossypol bound within the meal. Both solvent systems were effective at reducing gossypo...

  1. Comparison of automated nucleic acid extraction methods for the detection of cytomegalovirus DNA in fluids and tissues

    PubMed Central

    Waggoner, Jesse J.

    2014-01-01

    Testing for cytomegalovirus (CMV) DNA is increasingly being used for specimen types other than plasma or whole blood. However, few studies have investigated the performance of different nucleic acid extraction protocols in such specimens. In this study, CMV extraction using the Cell-free 1000 and Pathogen Complex 400 protocols on the QIAsymphony Sample Processing (SP) system were compared using bronchoalveolar lavage fluid (BAL), tissue samples, and urine. The QIAsymphonyAssay Set-up (AS) system was used to assemble reactions using artus CMV PCR reagents and amplification was carried out on the Rotor-Gene Q. Samples from 93 patients previously tested for CMV DNA and negative samples spiked with CMV AD-169 were used to evaluate assay performance. The Pathogen Complex 400 protocol yielded the following results: BAL, sensitivity 100% (33/33), specificity 87% (20/23); tissue, sensitivity 100% (25/25), specificity 100% (20/20); urine, sensitivity 100% (21/21), specificity 100% (20/20). Cell-free 1000 extraction gave comparable results for BAL and tissue, however, for urine, the sensitivity was 86% (18/21) and specimen quantitation was inaccurate. Comparative studies of different extraction protocols and DNA detection methods in body fluids and tissues are needed, as assays optimized for blood or plasma will not necessarily perform well on other specimen types. PMID:24765569

  2. Superheated water extraction of glycyrrhizic acid from licorice root.

    PubMed

    Shabkhiz, Mohammad A; Eikani, Mohammad H; Bashiri Sadr, Zeinolabedin; Golmohammad, Fereshteh

    2016-11-01

    Superheated water extraction (SWE) has become an interesting green extraction method for different classes of compounds. In this study, SWE was used to extract glycyrrhizic acid (GA) from licorice root. Response surface methodology (RSM) was applied to evaluate and optimize the extraction conditions. The influence of operating conditions such as water temperature (100, 120 and 140°C) and solvent flow rates (1, 3 and 5mL/min) were investigated at 0.5mm mean particle size and 20bar pressure. Separation and identification of the glycyrrhizic acid, as the main component, was carried out by the RP-HPLC method. The best operating conditions for the SWE of licorice were determined to be 100°C temperature,15mL/min flow rate and 120min extraction time. The results showed that the amount of the obtained GA was relatively higher using SWE (54.760mg/g) than the Soxhlet method (28.760mg/g) and ultrasonic extraction (18.240mg/g).

  3. A Novel Protocol to Analyze Short- and Long-Chain Fatty Acids Using Nonaqueous Microchip Capillary Electrophoresis

    NASA Technical Reports Server (NTRS)

    Cable, M. L.; Stockton, A. M.; Mora, Maria F; Willis, P. A.

    2013-01-01

    We propose a new protocol to identify and quantify both short- and long-chain saturated fatty acids in samples of astrobiological interest using non-aqueous microchip capillary electrophoresis (micronNACE) with laser induced fluorescence (LIF).

  4. Propionic acid production by extractive fermentation. 1. Solvent considerations

    SciTech Connect

    Gu, Z.; Glatz, B.A.; Glatz, C.E.

    1998-02-20

    Solvent selection for extractive fermentation for propionic acid was conducted with three systems: Alamine{reg_sign} 304-1 (trilaurylamine) in 2-octanol, 1-dodecanol, and Witcohol{reg_sign} 85 NF (oleyl alcohol). Among them, the solvent containing 2-octanol exhibited the highest partition coefficient in acid extraction, but it was also toxic to propionibacteria. The most solvent-resistant strain among five strains of the microorganism was selected. Solvent toxicity was eliminated via two strategies: entrapment of dissolved toxic solvent in the culture growth medium with vegetable oils such as corn, olive, or soybean oils; or replacement of the toxic 2-octanol with nontoxic Witcohol 85 NF. The complete recovery of acids from the Alamine 304-1/Witcohol 85 NF was also realized with vacuum distillation.

  5. Propionic acid production by extractive fermentation. I. Solvent considerations.

    PubMed

    Gu, Z; Glatz, B A; Glatz, C E

    1998-02-20

    Solvent selection for extractive fermentation for propionic acid was conducted with three systems: Alamine 304-1 (trilaurylamine) in 2-octanol, 1-dodecanol, and Witcohol 85 NF (oleyl alcohol). Among them, the solvent containing 2-octanol exhibited the highest partition coefficient in acid extraction, but it was also toxic to propionibacteria. The most solvent-resistant strain among five strains of the microorganism was selected. Solvent toxicity was eliminated via two strategies: entrapment of dissolved toxic solvent in the culture growth medium with vegetable oils such as corn, olive, or soybean oils; or replacement of the toxic 2-octanol with nontoxic Witcohol 85 NF. The complete recovery of acids from the Alamine 304-1/Witcohol 85 NF was also realized with vacuum distillation.

  6. Recovery of heavy metals from industrial sludge using various acid extraction approaches.

    PubMed

    Wu, C H; Kuo, C Y; Lo, S L

    2009-01-01

    Heavy metals were removed from industrial sludge by traditional and microwave-assisted acid extraction approaches. The effects of acid concentration, extraction time, sludge particle size and solid/liquid (S/L) ratio on copper-extraction efficiency were assessed. Leaching with increased acid concentrations increased the yield of heavy metals from the industrial sludge. In microwave-assisted acid extraction, reducing the S/L ratio and sludge particle size increased copper-extraction efficiency. These experimental findings indicate that S/L ratio most strongly influenced microwave-assisted acid extraction. Both traditional and microwave-assisted acid extraction demonstrate that sulfuric acid was an effective extractant, and the copper fraction in extracted sludge shifted from being primarily bound to Fe-Mn oxides and organic-matter partition, to being mostly bound to organic matter, remaining as a residue during acid extraction.

  7. Qualitative and quantitative evaluation of protein extraction protocols for apple and strawberry fruit suitable for two-dimensional electrophoresis and mass spectrometry analysis.

    PubMed

    Zheng, Qifa; Song, Jun; Doncaster, Kristen; Rowland, Elden; Byers, David M

    2007-03-07

    A modified phenol-based protocol and a phenol-free protocol that involves hot SDS extraction followed by TCA precipitation in acetone were qualitatively and quantitatively compared and evaluated on apple peel and strawberry fruit. The phenol protocol resulted in significantly higher protein yields of 2.35 +/- 0.1 and 0.46 +/- 0.06 mg/g of FW from apple and strawberry fruit, respectively, compared to the SDS protocol, which produced 0.74 +/- 0.1 and 0.27 +/- 0.02 mg/g of FW, respectively. 2-DE analysis of apple protein extracts revealed 1422 protein spots associated with the phenol protocol and 849 spots associated with the SDS protocol. Of these, 761 were present only in phenol gels, whereas 23 were exclusive to SDS samples. For strawberry, SDS extraction produced poor-quality spots with a high degree of streaking, indicating possible contamination. The application of a cleanup procedure resulted in a purified protein extract with high-quality spots. 2-DE analysis of strawberry protein extracts revealed 1368 spots for the phenol protocol and 956 spots for the SDS protocol accompanied by the cleanup procedure. Of these, 599 spots were present only in phenol gels, whereas 109 were present only in SDS samples. Spots from each fruit tissue and extraction procedure were selected, and a total of 26 were identified by LC-MS/MS. Overall, this study demonstrates the complexity of protein extraction of fruit tissues and suggests that a phenol-based protein extraction protocol should be used as a standard procedure for recalcitrant fruit tissues, whereas a SDS protocol with or without a cleanup procedure may be used as an alternative protocol.

  8. Improved proteomic analysis following trichloroacetic acid extraction of Bacillus anthracis spore proteins.

    PubMed

    Deatherage Kaiser, Brooke L; Wunschel, David S; Sydor, Michael A; Warner, Marvin G; Wahl, Karen L; Hutchison, Janine R

    2015-11-01

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Analysis of cellular proteins is dependent upon efficient extraction from bacterial samples, which can be challenging with increasing complexity and refractory characteristics. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrichment for certain classes of proteins. The method presented here is technically simple, does not require specialized equipment such as a mechanical disrupter, and is effective for protein extraction of the particularly challenging sample type of Bacillus anthracis Sterne spores. The ability of Trichloroacetic acid (TCA) extraction to isolate proteins from spores and enrich for spore-specific proteins was compared to the traditional mechanical disruption method of bead beating. TCA extraction improved the total average number of proteins identified within a sample as compared to bead beating (547 vs 495, respectively). Further, TCA extraction enriched for 270 spore proteins, including those typically identified by first isolating the spore coat and exosporium layers. Bead beating enriched for 156 spore proteins more typically identified from whole spore proteome analyses. The total average number of proteins identified was equal using TCA or bead beating for easily lysed samples, such as B. anthracis vegetative cells. As with all assays, supplemental methods such as implementation of an alternative preparation method may simplify sample preparation and provide additional insight to the protein biology of the organism being studied.

  9. Improved Proteomic Analysis Following Trichloroacetic Acid Extraction of Bacillus anthracis Spore Proteins

    SciTech Connect

    Kaiser, Brooke LD; Wunschel, David S.; Sydor, Michael A.; Warner, Marvin G.; Wahl, Karen L.; Hutchison, Janine R.

    2015-08-07

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Proteomic analysis is dependent upon efficient extraction of proteins from bacterial samples without introducing bias toward extraction of particular protein classes. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrich for certain classes of proteins. The method presented here is technically simple and does not require specialized equipment such as a mechanical disrupter. Our data reveal that for particularly challenging samples, such as B. anthracis Sterne spores, trichloroacetic acid extraction improved the number of proteins identified within a sample compared to bead beating (714 vs 660, respectively). Further, TCA extraction enriched for 103 known spore specific proteins whereas bead beating resulted in 49 unique proteins. Analysis of C. botulinum samples grown to 5 days, composed of vegetative biomass and spores, showed a similar trend with improved protein yields and identification using our method compared to bead beating. Interestingly, easily lysed samples, such as B. anthracis vegetative cells, were equally as effectively processed via TCA and bead beating, but TCA extraction remains the easiest and most cost effective option. As with all assays, supplemental methods such as implementation of an alternative preparation method may provide additional insight to the protein biology of the bacteria being studied.

  10. Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils

    PubMed Central

    Quideau, Sylvie A.; McIntosh, Anne C.S.; Norris, Charlotte E.; Lloret, Emily; Swallow, Mathew J.B.; Hannam, Kirsten

    2016-01-01

    Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors. The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis. PMID:27685177

  11. Extraction of Oleic Acid from Moroccan Olive Mill Wastewater

    PubMed Central

    Elkacmi, Reda; Kamil, Noureddine; Bennajah, Mounir; Kitane, Said

    2016-01-01

    The production of olive oil in Morocco has recently grown considerably for its economic and nutritional importance favored by the country's climate. After the extraction of olive oil by pressing or centrifuging, the obtained liquid contains oil and vegetation water which is subsequently separated by decanting or centrifugation. Despite its treatment throughout the extraction process, this olive mill wastewater, OMW, still contains a very important oily residue, always regarded as a rejection. The separated oil from OMW can not be intended for food because of its high acidity of 3.397% which exceeds the international standard for human consumption defined by the standard of the Codex Alimentarius, proving its poor quality. This work gives value addition to what would normally be regarded as waste by the extraction of oleic acid as a high value product, using the technique of inclusion with urea for the elimination of saturated and unsaturated fatty acids through four successive crystallizations at 4°C and 20°C to have a final phase with oleic acid purity of 95.49%, as a biodegradable soap and a high quality glycerin will be produced by the reaction of saponification and transesterification. PMID:26933663

  12. Comparison of Different Sample Preparation Protocols Reveals Lysis Buffer-Specific Extraction Biases in Gram-Negative Bacteria and Human Cells.

    PubMed

    Glatter, Timo; Ahrné, Erik; Schmidt, Alexander

    2015-11-06

    We evaluated different in-solution and FASP-based sample preparation strategies for absolute protein quantification. Label-free quantification (LFQ) was employed to compare different sample preparation strategies in the bacterium Pseudomonas aeruginosa and human embryonic kidney cells (HEK), and organismal-specific differences in general performance and enrichment of specific protein classes were noted. The original FASP protocol globally enriched for most proteins in the bacterial sample, whereas the sodium deoxycholate in-solution strategy was more efficient with HEK cells. Although detergents were found to be highly suited for global proteome analysis, higher intensities were obtained for high-abundant nucleic acid-associated protein complexes, like the ribosome and histone proteins, using guanidine hydrochloride. Importantly, we show for the first time that the observable total proteome mass of a sample strongly depends on the sample preparation protocol, with some protocols resulting in a significant underestimation of protein mass due to incomplete protein extraction of biased protein groups. Furthermore, we demonstrate that some of the observed abundance biases can be overcome by incorporating a nuclease treatment step or, alternatively, a correction factor for complementary sample preparation approaches.

  13. Protein biomarkers of external root resorption: A new protein extraction protocol. Are we going in the right direction?

    PubMed Central

    Vieira, Giovanni Modesto

    2014-01-01

    OBJECTIVE: The aim of this study is to determine a protocol of gingival crevicular fluid protein extraction used for the first dimension of 2-DE gels. It also aims at conducting a review on the current candidates for protein markers of this pathology, all of which may be used to prevent the disease. METHODS: Gingival crevicular fluid was collected from two groups of 60 patients each, with and without external root resorption. Samples were extracted by means of various methods of protein extraction. SDS-PAGE gels were used to assess the quality of the method which was subsequently tested during isoelectric focusing of 2-DE gels taken from samples of patients with and without the disease. RESULTS: Milli-Q ultrapure ice cold water, without precipitation for gingival crevicular fluid protein extraction, proved the method with greatest sharpness to detect protein bands. Additionally, it allowed two-dimensional electrophoresis to be performed. CONCLUSION: The new protein extraction protocol does not interfere in isoeletric focusing of 2-DE gels. Furthermore, it provides the greatest sharpness in detecting protein bands of SDS-PAGE gels. This will allow mapping and searching of new external root resorption markers, particularly due to the difficulty in carrying out molecular tests with the current candidates for protein markers. PMID:25628081

  14. Evaluation of five protocols for DNA extraction from leaves of Malus sieversii, Vitis vinifera, and Armeniaca vulgaris.

    PubMed

    Aubakirova, K; Omasheva, M; Ryabushkina, N; Tazhibaev, T; Kampitova, G; Galiakparov, N

    2014-02-27

    Leaves of Malus sieversii, Vitis vinifera, and Armeniaca vulgaris contain substantial amounts of secondary metabolites, which limit the high-quality DNA extraction performance. In this study, five extraction protocols were compared for their ability to produce good quality DNA from fresh and dried (with silica gel) leaves of these species. The modified protocol of Dellaporta et al., using polyvinylpyrrolidone to bind the phenolic compounds and a high molar concentration of potassium acetate to inhibit co-precipitation of polysaccharides with DNA, produced the best DNA quality for all species tested. DNA extracted by this method had a 1.77-1.96 A260/280 nm ratio and successful amplification of the 18S ribosomal DNA gene. DNA concentrations of dried leaves were lower than those obtained from fresh leaves, which was likely due to aspects of the drying procedure. All five methods for grapevine produced DNA of obvious better quality from green canes compared to leaves, due to the relatively low content of secondary metabolites in the former. For grapevine and apricot, three methods can be equally used to obtain DNA of good quality: the Doyle and Doyle modified method using CTAB and high concentration of NaCl, the Jobes et al. modified method, and the sodium dodecyl sulfate mini preparation method of Edwards et al. The protocol of Jobes et al. using LiCl for RNA removal showed the best results for most of the M. sieversii samples examined.

  15. Extraction of nitric acid, uranyl nitrate, and bismuth nitrate from aqueous nitric acid solutions with CMPO

    SciTech Connect

    Spencer, B.B.

    1995-08-01

    DOE sponsored development of the transuranium extraction (TRUEX) process for removing actinides from radioactive wastes. The solvent is a mixture of CMPO and TBP. Since the extraction characteristics of CMPO are not as well understood as those of TBP, the extraction of nitric acid, uranyl nitrate, and bismuth nitrate with CMPO (dissolved in n-dodecane) were studied. Results indicate that CMPO extracts nitric acid with a 1:1 stoichiometry; equilibrium constant is 2. 660{plus_minus}0.092 at 25 C, and extraction enthalpy is -5. 46{plus_minus}0.46 kcal/mol. Slope analysis indicates that uranyl nitrate extracts with a mixed equilibria of 1:1 and 2:1 stoichiometries in nearly equal proportion. Equil. constant of the 2: 1 extraction was 1.213 {times} 10{sup 6}{plus_minus}3.56 {times} 10{sup 4} at 25 C; reaction enthalpy was -9.610{plus_minus}0.594 kcal/mol. Nitration complexation constant is 8.412{plus_minus}0.579, with an enthalpy of -10.72{plus_minus}1.87 kcal/mol. Bismuth nitrate also extracts with a mixed equilibria of (perhaps) 1:1 and 2:1 stoichiometries. A 2:1 extraction equilibrium and a nitrate complexation adequately model the data. Kinetics and enthalpies were also measured.

  16. Hydrothermal acid treatment for sugar extraction from Golenkinia sp.

    PubMed

    Choi, Sun-A; Choi, Won-Il; Lee, Jin-Suk; Kim, Seung Wook; Lee, Gye-An; Yun, Jihyun; Park, Ji-Yeon

    2015-08-01

    In this study, hydrothermal acid treatment for efficient recovery of sugar from Golenkinia sp. was investigated. The initial glucose and XMG (xylose, mannose, and galactose) contents of a prepared Golenkinia sp. solution (40g/L) were 15.05 and 5.24g/L, respectively. The microalgal cell walls were hydrolyzed, for sugar recovery, by enzymatic saccharification and/or hydrothermal acid treatment. Among the various hydrothermal acid treatment conditions, the most optimal were the 2.0% H2SO4 concentration at 150°C for 15min, under which the glucose- and XMG-extraction yields were 71.7% and 64.9%, respectively. By pH 4.8, 50°C enzymatic hydrolysis after optimal hydrothermal acid treatment, the glucose- and XMG-extraction yields were additionally increased by 8.3% and 0.8%, respectively. After hydrothermal acid treatment, the combination with the enzymatic hydrolysis process improved the total sugar yield of Golenkinia sp. to 75.4%.

  17. Optimizing preservation protocols to extract high-quality RNA from different tissues of echinoderms for next-generation sequencing.

    PubMed

    Pérez-Portela, Rocío; Riesgo, Ana

    2013-09-01

    Transcriptomic information provides fundamental insights into biological processes. Extraction of quality RNA is a challenging step, and preservation and extraction protocols need to be adjusted in many cases. Our objectives were to optimize preservation protocols for isolation of high-quality RNA from diverse echinoderm tissues and to compare the utility of parameters as absorbance ratios and RIN values to assess RNA quality. Three different tissues (gonad, oesophagus and coelomocytes) were selected from the sea urchin Arbacia lixula. Solid tissues were flash-frozen and stored at -80 °C until processed. Four preservation treatments were applied to coelomocytes: flash freezing and storage at -80 °C, RNAlater and storage at -20 °C, preservation in TRIzol reagent and storage at -80 °C and direct extraction with TRIzol from fresh cells. Extractions of total RNA were performed with a modified TRIzol protocol for all tissues. Our results showed high values of RNA quantity and quality for all tissues, showing nonsignificant differences among them. However, while flash freezing was effective for solid tissues, it was inadequate for coelomocytes because of the low quality of the RNA extractions. Coelomocytes preserved in RNAlater displayed large variability in RNA integrity and insufficient RNA amount for further isolation of mRNA. TRIzol was the most efficient system for stabilizing RNA which resulted on high RNA quality and quantity. We did not detect correlation between absorbance ratios and RNA integrity. The best strategies for assessing RNA integrity was the visualization of 18S rRNA and 28S rRNA bands in agarose gels and estimation of RIN values with Agilent Bioanalyzer chips.

  18. Lactic acid bacterial extract as a biogenic mineral growth modifier

    NASA Astrophysics Data System (ADS)

    Borah, Ballav M.; Singh, Atul K.; Ramesh, Aiyagari; Das, Gopal

    2009-04-01

    The formation of minerals and mechanisms by which bacteria could control their formation in natural habitats is now of current interest for material scientists to have an insight of the mechanism of in vivo mineralization, as well as to seek industrial and technological applications. Crystalline uniform structures of calcium and barium minerals formed micron-sized building blocks when synthesized in the presence of an organic matrix consisting of secreted protein extracts from three different lactic acid bacteria (LAB) viz.: Lactobacillus plantarum MTCC 1325, Lactobacillus acidophilus NRRL B4495 and Pediococcus acidilactici CFR K7. LABs are not known to form organic matrix in biological materialization processes. The influence of these bacterial extracts on the crystallization behavior was investigated in details to test the basic coordination behavior of the acidic protein. In this report, varied architecture of the mineral crystals obtained in presence of high molecular weight protein extracts of three different LAB strains has been discussed. The role of native form of high molecular weight bacterial protein extracts in the generation of nucleation centers for crystal growth was clearly established. A model for the formation of organic matrix-cation complex and the subsequent events leading to crystal growth is proposed.

  19. Unusual stable isotope ratios in amino acid and carboxylic acid extracts from the Murchison meteorite

    NASA Technical Reports Server (NTRS)

    Epstein, S.; Krishnamurthy, R. V.; Cronin, J. R.; Pizzarello, S.; Yuen, G. U.

    1987-01-01

    The isotopic composition of hydrogen, nitrogen, and carbon in amino acid and monocarboxylic acid extracts from the Murchison meteorite has been determined. The unusually high D/H and N-15/N-14 ratios in the amino acid fraction are uniquely characteristic of known interstellar organic materials. The delta D value of the monocarboxylic acid fraction is lower but still consistent with an interstellar origin. These results confirm the extraterrestrial origin of both classes of compound and provide the first evidence suggesting a direct relationship between the massive organosynthesis occurring in interstellar clouds and the presence of prebiotic compounds in primitive planetary bodies.

  20. RNA extraction from various recalcitrant plant tissues with a cethyltrimethylammonium bromide-containing buffer followed by an acid guanidium thiocyanate-phenol-chloroform treatment.

    PubMed

    Suzuki, Yuji; Mae, Tadahiko; Makino, Amane

    2008-07-01

    High-quality total RNA was extracted using a cethyltrimethylammonium bromide-containing buffer followed by an acid guanidium thiocyanate-phenol-chloroform treatment from recalcitrant plant tissues such as tree leaves (pine, Norway spruce, ginkgo, Japanese cedar, rose), flowers (rose, Lotus japonicus) and storage tissues (seeds of Lotus japonicus and rice, sweet potato tuber, banana fruit). This protocol greatly reduced the time required for RNA extraction.

  1. Estimates of Soil Bacterial Ribosome Content and Diversity Are Significantly Affected by the Nucleic Acid Extraction Method Employed.

    PubMed

    Wüst, Pia K; Nacke, Heiko; Kaiser, Kristin; Marhan, Sven; Sikorski, Johannes; Kandeler, Ellen; Daniel, Rolf; Overmann, Jörg

    2016-05-01

    Modern sequencing technologies allow high-resolution analyses of total and potentially active soil microbial communities based on their DNA and RNA, respectively. In the present study, quantitative PCR and 454 pyrosequencing were used to evaluate the effects of different extraction methods on the abundance and diversity of 16S rRNA genes and transcripts recovered from three different types of soils (leptosol, stagnosol, and gleysol). The quality and yield of nucleic acids varied considerably with respect to both the applied extraction method and the analyzed type of soil. The bacterial ribosome content (calculated as the ratio of 16S rRNA transcripts to 16S rRNA genes) can serve as an indicator of the potential activity of bacterial cells and differed by 2 orders of magnitude between nucleic acid extracts obtained by the various extraction methods. Depending on the extraction method, the relative abundances of dominant soil taxa, in particular Actino bacteria and Proteobacteria, varied by a factor of up to 10. Through this systematic approach, the present study allows guidelines to be deduced for the selection of the appropriate extraction protocol according to the specific soil properties, the nucleic acid of interest, and the target organisms.

  2. Estimates of Soil Bacterial Ribosome Content and Diversity Are Significantly Affected by the Nucleic Acid Extraction Method Employed

    PubMed Central

    Wüst, Pia K.; Nacke, Heiko; Kaiser, Kristin; Marhan, Sven; Sikorski, Johannes; Kandeler, Ellen; Daniel, Rolf

    2016-01-01

    Modern sequencing technologies allow high-resolution analyses of total and potentially active soil microbial communities based on their DNA and RNA, respectively. In the present study, quantitative PCR and 454 pyrosequencing were used to evaluate the effects of different extraction methods on the abundance and diversity of 16S rRNA genes and transcripts recovered from three different types of soils (leptosol, stagnosol, and gleysol). The quality and yield of nucleic acids varied considerably with respect to both the applied extraction method and the analyzed type of soil. The bacterial ribosome content (calculated as the ratio of 16S rRNA transcripts to 16S rRNA genes) can serve as an indicator of the potential activity of bacterial cells and differed by 2 orders of magnitude between nucleic acid extracts obtained by the various extraction methods. Depending on the extraction method, the relative abundances of dominant soil taxa, in particular Actinobacteria and Proteobacteria, varied by a factor of up to 10. Through this systematic approach, the present study allows guidelines to be deduced for the selection of the appropriate extraction protocol according to the specific soil properties, the nucleic acid of interest, and the target organisms. PMID:26896137

  3. Ultrasound-assisted extraction of carnosic acid and rosmarinic acid using ionic liquid solution from Rosmarinus officinalis.

    PubMed

    Zu, Ge; Zhang, Rongrui; Yang, Lei; Ma, Chunhui; Zu, Yuangang; Wang, Wenjie; Zhao, Chunjian

    2012-01-01

    Ionic liquid based, ultrasound-assisted extraction was successfully applied to the extraction of phenolcarboxylic acids, carnosic acid and rosmarinic acid, from Rosmarinus officinalis. Eight ionic liquids, with different cations and anions, were investigated in this work and [C(8)mim]Br was selected as the optimal solvent. Ultrasound extraction parameters, including soaking time, solid-liquid ratio, ultrasound power and time, and the number of extraction cycles, were discussed by single factor experiments and the main influence factors were optimized by response surface methodology. The proposed approach was demonstrated as having higher efficiency, shorter extraction time and as a new alternative for the extraction of carnosic acid and rosmarinic acid from R. officinalis compared with traditional reference extraction methods. Ionic liquids are considered to be green solvents, in the ultrasound-assisted extraction of key chemicals from medicinal plants, and show great potential.

  4. Ultrasound-Assisted Extraction of Carnosic Acid and Rosmarinic Acid Using Ionic Liquid Solution from Rosmarinus officinalis

    PubMed Central

    Zu, Ge; Zhang, Rongrui; Yang, Lei; Ma, Chunhui; Zu, Yuangang; Wang, Wenjie; Zhao, Chunjian

    2012-01-01

    Ionic liquid based, ultrasound-assisted extraction was successfully applied to the extraction of phenolcarboxylic acids, carnosic acid and rosmarinic acid, from Rosmarinus officinalis. Eight ionic liquids, with different cations and anions, were investigated in this work and [C8mim]Br was selected as the optimal solvent. Ultrasound extraction parameters, including soaking time, solid–liquid ratio, ultrasound power and time, and the number of extraction cycles, were discussed by single factor experiments and the main influence factors were optimized by response surface methodology. The proposed approach was demonstrated as having higher efficiency, shorter extraction time and as a new alternative for the extraction of carnosic acid and rosmarinic acid from R. officinalis compared with traditional reference extraction methods. Ionic liquids are considered to be green solvents, in the ultrasound-assisted extraction of key chemicals from medicinal plants, and show great potential. PMID:23109836

  5. Stabilization of caffeic acid derivatives in Echinacea purpurea L. glycerin extract.

    PubMed

    Bergeron, Chantal; Gafner, Stefan; Batcha, Laura L; Angerhofer, Cindy K

    2002-07-03

    Recent work has shown that enzymatic degradation and oxidation of cichoric acid and other caffeic derivatives occurs in Echinacea preparations. However, very little is known as to the means of stabilizing these phytopreparations. To stabilize the glycerin extract of Echinacea purpurea, we have evaluated the effects of 3 natural antioxidants (citric acid, malic acid, and hibiscus extract) on the stability of the major caffeic acid derivatives (caftaric acid, caffeic acid, cichoric acid, and 2-O-feruloyl-tartaric acid). Chlorogenic acid, which normally occurs in an ethanol extract of E. purpurea, was not present in the glycerin extract. The caffeic acid derivatives, with the exception of 2-O-feruloyl-tartaric acid, were subject to degradation in the control sample. 2-O-Feruloyl-tartaric acid was stable during the whole testing period. All antioxidant treatments greatly improved the stability of caffeic acid derivatives. Stability was dependent upon the concentration of antioxidant added.

  6. Development of a protocol for determination of domoic acid in the sand crab (Emerita analoga): a possible new indicator species.

    PubMed

    Powell, Christine L; Ferdin, M E; Busman, Mark; Kvitek, Rikk G; Doucette, Gregory J

    2002-05-01

    The aim of this study was to begin evaluating the utility of sand crabs (Emerita analoga) as an indicator species for the algal neurotoxin, domoic acid (DA), in Monterey Bay, California, USA, a site of recurrent blooms of the DA-producing diatom, Pseudo-nitzschia. One of the current sentinel organisms, the sea mussel (Mytilus californianus), has shown minimal or undetectable toxicity during some local bloom events. As a critical step in assuring the accuracy of DA determinations in E. analoga, we have developed and validated a highly efficient extraction protocol that yields toxin recoveries of 97+/-2.9%. We also determined by HPLC-UV and receptor binding assay, with confirmation by LC-MS/MS, that sand crabs accumulated measurable amounts of DA during toxic Pseudo-nitzschia blooms, while the sea mussel showed no detectable toxin. In addition, a comparison of inter-animal variability in DA content revealed values ranging from ca. 0.5 to 5 microg DAg(-1) tissue and no consistent trend with size class, based on either animal weight or length. These data on the toxicity of individual animals will be useful in designing an appropriate sampling strategy for monitoring DA and, importantly, indicate that sand crabs do not appear to progressively bioaccumulate DA with age.

  7. Rapid DNA extraction protocol for detection of alpha-1 antitrypsin deficiency from dried blood spots by real-time PCR.

    PubMed

    Struniawski, R; Szpechcinski, A; Poplawska, B; Skronski, M; Chorostowska-Wynimko, J

    2013-01-01

    The dried blood spot (DBS) specimens have been successfully employed for the large-scale diagnostics of α1-antitrypsin (AAT) deficiency as an easy to collect and transport alternative to plasma/serum. In the present study we propose a fast, efficient, and cost effective protocol of DNA extraction from dried blood spot (DBS) samples that provides sufficient quantity and quality of DNA and effectively eliminates any natural PCR inhibitors, allowing for successful AAT genotyping by real-time PCR and direct sequencing. DNA extracted from 84 DBS samples from chronic obstructive pulmonary disease patients was genotyped for AAT deficiency variants by real-time PCR. The results of DBS AAT genotyping were validated by serum IEF phenotyping and AAT concentration measurement. The proposed protocol allowed successful DNA extraction from all analyzed DBS samples. Both quantity and quality of DNA were sufficient for further real-time PCR and, if necessary, for genetic sequence analysis. A 100% concordance between AAT DBS genotypes and serum phenotypes in positive detection of two major deficiency S- and Z- alleles was achieved. Both assays, DBS AAT genotyping by real-time PCR and serum AAT phenotyping by IEF, positively identified PI*S and PI*Z allele in 8 out of the 84 (9.5%) and 16 out of 84 (19.0%) patients, respectively. In conclusion, the proposed protocol noticeably reduces the costs and the hand-on-time of DBS samples preparation providing genomic DNA of sufficient quantity and quality for further real-time PCR or genetic sequence analysis. Consequently, it is ideally suited for large-scale AAT deficiency screening programs and should be method of choice.

  8. Antioxidant activity and sensory evaluation of a rosmarinic acid-enriched extract of Salvia officinalis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An extract of S. officinalis (garden sage) was developed using supercritical fluid extraction, followed by hot water extraction. The resulting extract was enriched in polyphenols, including rosmarinic acid (RA), which has shown promising health benefits in animals. The extract contained RA at a conc...

  9. Extraction Behaviors of Heavy Rare Earths with Organophosphoric Extractants: The Contribution of Extractant Dimer Dissociation, Acid Ionization, and Complexation. A Quantum Chemistry Study.

    PubMed

    Jing, Yu; Chen, Ji; Chen, Li; Su, Wenrou; Liu, Yu; Li, Deqian

    2017-03-30

    Heavy rare earths (HREs), namely Ho(3+), Er(3+), Tm(3+), Yb(3+) and Lu(3+), are rarer and more exceptional than light rare earths, due to the stronger extraction capacity for 100 000 extractions. Therefore, their incomplete stripping and high acidity of stripping become problems for HRE separation by organophosphoric extractants. However, the theories of extractant structure-performance relationship and molecular design method of novel HRE extractants are still not perfect. Beyond the coordination chemistry of the HRE-extracted complex, the extractant dimer dissociation, acid ionization, and complexation behaviors can be crucial to HRE extraction and reactivity of ionic species for understanding and further improving the extraction performance. To address the above issues, three primary fundamental processes, including extractant dimer dissociation, acid ionization, and HRE complexation, were identified and investigated systematically. The intrinsic extraction performances of HRE cations with four acidic organophosphoric extractants (P507, P204, P227 and Cyanex 272) were studied by using relativistic energy-consistent 4f core pseudopotentials, combined with density functional theory and a solvation model. Four acidic organophosphoric extractants have been qualified quantitatively from microscopic structures to chemical properties. It has been found that the Gibbs free energy changes of the overall extraction process (sequence: P204 > P227 > P507 > Cyanex 272) and their differences as a function of HREs (sequence: Ho/Er > Er/Tm > Tm/Yb > Yb/Lu) are in good agreement with the experimental maximum extraction capacities and separation factors. These results could provide an important approach to evaluate HRE extractants by the comprehensive consideration of dimer dissociation, acid ionization, and complexation processes. This paper also demonstrates the importance of the P-O bond, the P-C bond, isomer substituent, and solvation effects on the structure

  10. Method for extracting lanthanides and actinides from acid solutions

    DOEpatents

    Horwitz, E. Philip; Kalina, Dale G.; Kaplan, Louis; Mason, George W.

    1985-01-01

    A process for the recovery of actinide and lanthanide values from aqueous acidic solutions with an organic extractant having the formula: ##STR1## where .phi. is phenyl, R.sup.1 is a straight or branched alkyl or alkoxyalkyl containing from 6 to 12 carbon atoms and R.sup.2 is an alkyl containing from 3 to 6 carbon atoms. The process is suitable for the separation of actinide and lanthanide values from fission product values found together in high level nuclear reprocessing waste solutions.

  11. Evaluation and optimization of nucleic acid extraction methods for the molecular analysis of bacterial communities associated with corroded carbon steel.

    PubMed

    Marty, Florence; Ghiglione, Jean-François; Païssé, Sandrine; Gueuné, Hervé; Quillet, Laurent; van Loosdrecht, Mark C M; Muyzer, Gerard

    2012-01-01

    Different DNA and RNA extraction approaches were evaluated and protocols optimized on in situ corrosion products from carbon steel in marine environments. Protocols adapted from the PowerSoil DNA/RNA Isolation methods resulted in the best nucleic acid (NA) extraction performances (ie combining high NA yield, quality, purity, representativeness of microbial community and processing time efficiency). The PowerSoil RNA Isolation Kit was the only method which resulted in amplifiable RNA of good quality (ie intact 16S/23S rRNA). Sample homogenization and hot chemical (SDS) cell lysis combined with mechanical (bead-beating) lysis in presence of a DNA competitor (skim milk) contributed to improving substantially (around 23 times) the DNA yield of the PowerSoil DNA Isolation Kit. Apart from presenting NA extraction strategies for optimizing extraction parameters with corrosion samples from carbon steel, this study proposes DNA and RNA extraction procedures suited for comparative molecular analysis of total and active fractions of bacterial communities associated with carbon steel corrosion events, thereby contributing to improved MIC diagnosis and control.

  12. Calcium extraction from brine water and seawater using oxalic acid

    NASA Astrophysics Data System (ADS)

    Natasha, Nadia Chrisayu; Lalasari, Latifa Hanum

    2017-01-01

    Calcium can be extracted not only from rocks but also from natural liquor such as seawater and brine water. In order to extract the calcium from seawater and brine water, oxalic acid was used in this research. Effect of variations of the volume of the oxalic acid at a constant concentration in seawater and brine water to produce calcium was investigated. The concentration of oxalic acid was 100 g/l and the variations of its volume were 2 ml, 4 ml, 6 ml, 8 ml, 10 ml, 20 ml, 30 ml, 40 ml, and 50 ml. The used seawater and brine water were firstly evaporated from 100 ml into 50 ml and then the oxalic acid was added into them with mixing to produce the calcium precipitates. The precipitates were analyzed by X-ray diffraction (XRD) and scanning electron microscope (SEM) and the filtrates were analyzed by inductively coupled plasma-optical emission spectrometry (ICP-OES). The SEM analysis showed that the precipitates from brine water were consisted of only calcium compound while from seawater sodium one was also found along with calcium compound. The XRD analysis showed that the calcium was present in the form of calcium oxalate for both seawater and brine water. The ICP-OES analysis of the filtrate from seawater precipitation showed that the its calcium content was decreased from 826.20 ppm to 0.04 ppm while from brine water, it decreased from 170.06 ppm to 1.96 ppm. These results showed that both seawater and brine water have the potential to be a raw material for calcium production.

  13. Fast determination of bioactive phytic acid and pyrophosphate in walnuts using microwave accelerated extraction.

    PubMed

    Liu, Tong; He, Liu; Valiente, Manuel; López-Mesas, Montserrat

    2017-04-15

    Bioactive compounds phytic acid (IP6) and pyrophosphate (PPi) are minor components of walnuts with the ability of being inhibitors of urolithiasis, among others. Since simultaneous analysis of IP6 and PPi have known drawbacks, a new method to determine their content in walnuts has been developed with emphasis on their extraction from walnuts by microwave-assisted extraction (MAE). Acid content of extracting solvent, extraction time and temperature were optimized. After extraction, compounds were purified by selective adsorption/desorption on an anion exchange solid phase extraction and analyzed by inductive coupled plasma/mass spectrometry. A mixture of H2SO4 and HCl as solvent to extract both, IP6 and PPi, provided results slightly higher than those determined by conventional extraction with no statistical difference. The possible hydrolysis of phytic acid by MAE was analyzed. Compared with the conventional acid extraction method, significant improvement is achieved by the MAE method reducing extraction time from 3h to 10min.

  14. Extraction of Palladium from Nitric Acid by Diamides of Di-picolinic Acid

    SciTech Connect

    Alyapyshev, M.Yu.; Babain, V.A.; Pokhitonov, Yu.A.; Esimantovskiy, V.M.

    2007-07-01

    The most complicated and urgent problem of atomic industry consists in the safe isolation and storage of radioactive wastes. The long-lived radionuclides presented in high-level liquid wastes (HLLW) pose a potential threat to environment for hundreds and thousands of years. One of the possible ways to reduce the danger of HLLW storages is concerned with treatment of HLLW intended to recovery of long-lived radionuclides and their partitioning into separate fractions. The separation of the most hazardous radionuclides (like transplutonium elements (TPE)) to the individual fraction of low volume leads to decrease of the total volume of HLLW and therefore to decrease of solidified waste storage costs. It should be noted that only in the case of reprocessing it can be possible to recover individual radionuclides (or their fractions) into separate flows with further special approach to each of them. Partitioning of different HLLW is under investigation in many countries now. Numerous processes for recovery of Cs, Sr, TPE and REE have been already developed and tested. At the same time partitioning is only the first step on the road to the following synthesis of materials providing the safe storage of long-lived radionuclides over many thousands of years. The metallic palladium contained in HLLW seems to be a promising material for producing of matrices for incorporation of radioactive wastes. Different methods for palladium recovery have been investigated: reductive precipitation, electrochemical precipitation, sorption and extraction. Of prime importance are extraction methods. Phosphine oxides, carbamoyl-phosphine oxides, crown-ethers, oximes, sulfides and some other compounds were proposed as extractants towards palladium from nitric acid media. It is reasonable to recover palladium into individual fraction during waste partitioning. Diamides of malonic, di-glycolic and pyridine-dicarboxylic (di-picolinic) acids are intensively investigated as extractants for HLLW

  15. Enantioselective liquid-liquid extraction of zopiclone with mandelic acid ester derivatives.

    PubMed

    Peng, Yangfeng; He, Quan; Zuo, Bin; Niu, Haibo; Tong, Tianzhong; Zhao, Hongliang

    2013-12-01

    Enantioselective liquid-liquid extraction of zopiclone was conducted by employing a series of (R)-mandelic acid esters as chiral extractants. The effects of concentration of extractant, concentration of zopiclone, type of organic solvent, pH value, and temperature on the extraction efficiency were investigated. (R)-o-chloromandelic acid propyl ester was demonstrated to be an efficient chiral extractant for zopiclone resolution with a maximum enantioselectivity of 1.6.

  16. Trace DNA from insect skins: a comparison of five extraction protocols and direct PCR on chironomid pupal exuviae.

    PubMed

    Kranzfelder, Petra; Ekrem, Torbjørn; Stur, Elisabeth

    2016-01-01

    Insect skins (exuviae) are of extracellular origin and shed during moulting. The skins do not contain cells or DNA themselves, but epithelial cells and other cell-based structures might accidentally attach as they are shed. This source of trace DNA can be sufficient for PCR amplification and sequencing of target genes and aid in species identification through DNA barcoding or association of unknown life stages. Species identification is essential for biomonitoring programs, as species vary in sensitivities to environmental factors. However, it requires a DNA isolation protocol that optimizes the output of target DNA. Here, we compare the relative effectiveness of five different DNA extraction protocols and direct PCR in isolation of DNA from chironomid pupal exuviae. Chironomidae (Diptera) is a species-rich group of aquatic macroinvertebrates widely distributed in freshwater environments and considered a valuable bioindicator of water quality. Genomic DNA was extracted from 61.2% of 570 sampled pupal exuviae. There were significant differences in the methods with regard to cost, handling time, DNA quantity, PCR success, sequence success and the ability to sequence target taxa. The NucleoSpin(®) Tissue XS Kit, DNeasy(®) Blood and Tissue kit, and QuickExtract(™) DNA Extraction Solution provided the best results in isolating DNA from single pupal exuviae. Direct PCR and DTAB/CTAB methods gave poor results. While the observed differences in DNA isolation methods on trace DNA will be relevant to research that focuses on aquatic macroinvertebrate ecology, taxonomy and systematics, they should also be of interest for studies using environmental barcoding and metabarcoding of aquatic environments.

  17. Variables influencing extraction of nucleic acids from microbial plankton (viruses, bacteria, and protists) collected on nanoporous aluminum oxide filters.

    PubMed

    Mueller, Jaclyn A; Culley, Alexander I; Steward, Grieg F

    2014-07-01

    Anodic aluminum oxide (AAO) filters have high porosity and can be manufactured with a pore size that is small enough to quantitatively capture viruses. These properties make the filters potentially useful for harvesting total microbial communities from water samples for molecular analyses, but their performance for nucleic acid extraction has not been systematically or quantitatively evaluated. In this study, we characterized the flux of water through commercially produced nanoporous (0.02 μm) AAO filters (Anotop; Whatman) and used isolates (a virus, a bacterium, and a protist) and natural seawater samples to test variables that we expected would influence the efficiency with which nucleic acids are recovered from the filters. Extraction chemistry had a significant effect on DNA yield, and back flushing the filters during extraction was found to improve yields of high-molecular-weight DNA. Using the back-flush protocol, the mass of DNA recovered from microorganisms collected on AAO filters was ≥ 100% of that extracted from pellets of cells and viruses and 94% ± 9% of that obtained by direct extraction of a liquid bacterial culture. The latter is a minimum estimate of the relative recovery of microbial DNA, since liquid cultures include dissolved nucleic acids that are retained inefficiently by the filter. In conclusion, we demonstrate that nucleic acids can be extracted from microorganisms on AAO filters with an efficiency similar to that achievable by direct extraction of microbes in suspension or in pellets. These filters are therefore a convenient means by which to harvest total microbial communities from multiple aqueous samples in parallel for subsequent molecular analyses.

  18. Water-enhanced solubility of carboxylic acids in organic solvents and its applications to extraction processes

    SciTech Connect

    Starr, J.N.; King, C.J.

    1991-11-01

    The solubilities of carboxylic acids in certain organic solvents increase remarkably with an increasing amount of water in the organic phase. This phenomenon leads to a novel extract regeneration process in which the co-extracted water is selectively removed from an extract, and the carboxylic acid precipitates. This approach is potentially advantageous compared to other regeneration processes because it removes a minor component of the extract in order to achieve a large recovery of acid from the extract. Carboxylic acids of interest include adipic acid, fumaric acid, and succinic acid because of their low to moderate solubilities in organic solvents. Solvents were screened for an increase in acid solubility with increased water concentration in the organic phase. Most Lewis-base solvents were found to exhibit this increased solubility phenomena. Solvents that have a carbonyl functional group showed a very large increase in acid solubility. 71 refs., 52 figs., 38 tabs.

  19. Bacterial and fungal DNA extraction from positive blood culture bottles: a manual and an automated protocol.

    PubMed

    Mäki, Minna

    2015-01-01

    When adapting a gene amplification-based method in a routine sepsis diagnostics using a blood culture sample as a specimen type, a prerequisite for a successful and sensitive downstream analysis is the efficient DNA extraction step. In recent years, a number of in-house and commercial DNA extraction solutions have become available. Careful evaluation in respect to cell wall disruption of various microbes and subsequent recovery of microbial DNA without putative gene amplification inhibitors should be conducted prior selecting the most feasible DNA extraction solution for the downstream analysis used. Since gene amplification technologies have been developed to be highly sensitive for a broad range of microbial species, it is also important to confirm that the used sample preparation reagents and materials are bioburden-free to avoid any risks for false-positive result reporting or interference of the diagnostic process. Here, one manual and one automated DNA extraction system feasible for blood culture samples are described.

  20. A novel extraction protocol to probe the role of cholesterol in synaptic vesicle recycling.

    PubMed

    Dason, Jeffrey S; Charlton, Milton P

    2014-01-01

    Cholesterol helps to stabilize membrane fluidity and many membrane proteins interact with cholesterol and are functionally clustered in cholesterol rich "rafts." Synaptic vesicle (SV) membranes are enriched in cholesterol in comparison to other organelles. Attempts to study the function of this high cholesterol content have been hampered by the inability to extract cholesterol from SVs in live presynaptic terminals. Here, we describe a method to extract vesicular cholesterol using a temperature-sensitive Drosophila dynamin mutant, shibire-ts1 (shi), to trap SVs on the plasma membrane. Trapped SVs are more accessible to cholesterol extraction by the cholesterol chelator, methyl-β-cyclodextrin (MβCD). This method can likely be extended to extract other lipids from SVs and could also be used to add lipids. We speculate that this method could be used on mammalian preparations in conjunction with dynamin inhibitors.

  1. Evaluation of extraction protocols for anti-diabetic phytochemical substances from medicinal plants

    PubMed Central

    Okoduwa, Stanley Irobekhian Reuben; Umar, Ismaila A; James, Dorcas B; Inuwa, Hajara M; Habila, James D

    2016-01-01

    AIM To examine the efficacy of three extraction techniques: Soxhlet-extraction (SE), cold-maceration (CM) and microwave-assisted-extraction (MAE) using 80% methanol as solvent. METHODS The study was performed on each of 50 g of Vernonia amygdalina (VA) and Occimum gratissimum (OG) leaves respectively. The percentage yield, duration of extraction, volume of solvent used, qualitative and quantitative phytoconstituents present was compared. The biological activities (hypoglycemic effect) were investigated using albino wistar rat model of diabetes mellitus (n = 36) with a combined dose (1:1) of the two plants leaf extracts (250 mg/kg b.w.) from the three methods. The extracts were administered orally, once daily for 21 d. RESULTS In this report, the percentage VA extract yield from MAE was highest (20.9% ± 1.05%) within 39 min using 250 mL of solvent, when compared to the CM (14.35% ± 0.28%) within 4320 min using 900 mL of solvent and SE (15.75% ± 0.71%) within 265 min using 500 mL of solvent. The percentage differences in OG extract yield between: MAE vs SE was 41.05%; MAE vs CM was 46.81% and SE vs CM was 9.77%. The qualitative chemical analysis of the two plants showed no difference in the various phytoconstituents tested, but differs quantitatively in the amount of the individual phytoconstituents, as MAE had significantly high yield (P > 0.05) on phenolics, saponins and tannins. SE technique gave significantly high yield (P > 0.05) on alkaloid, while CM gave significant high yield on flavonoids. The extracts from CM exhibited a significantly (P > 0.05) better hypoglycemic activity within the first 14-d of treatment (43.3% ± 3.62%) when compared to MAE (36.5% ± 0.08%) and SE methods (33.3% ± 1.60%). However, the percentage hypoglycemic activity, 21 d post-treatment with 250 mg/kg b.w. extract from MAE was 72.6% ± 1.03% and it was more comparable to 10 mg/kg b.w. glibenclamide treated group (75.0% ± 0.73%), unlike the SE (69.5% ± 0.71%) and CM (69.1% ± 1

  2. Role of modifier in microwave assisted extraction of oleanolic acid from Gymnema sylvestre: application of green extraction technology for botanicals.

    PubMed

    Mandal, Vivekananda; Dewanjee, Saikat; Mandal, Subhash C

    2009-08-01

    This work highlights the development of a green extraction technology for botanicals with the use of microwave energy. Taking into consideration the extensive time involved in conventional extraction methods, coupled with usage of large volumes of organic solvent and energy resources, an ecofriendly green method that can overcome the above problems has been developed. The work compares the effect of sample pretreatment with untreated sample for improved yield of oleanolic acid from Gymnema sylvestre leaves. The pretreated sample with water produced 0.71% w/w oleanolic acid in one extraction cycle with 500 W microwave power, 25 mL methanol and only an 8 min extraction time. On the other hand, a conventional heat reflux extraction for 6 hours could produce only 0.62% w/w oleanolic acid. The detailed mechanism of extraction has been studied through scanning electron micrographs. The environmental impact of the proposed green method has also been evaluated.

  3. Comparison of DNA extraction protocols for microbial communities from soil treated with biochar

    PubMed Central

    Leite, D.C.A.; Balieiro, F.C.; Pires, C.A.; Madari, B.E.; Rosado, A.S.; Coutinho, H.L.C.; Peixoto, R.S.

    2014-01-01

    Many studies have evaluated the effects of biochar application on soil structure and plant growth. However, there are very few studies describing the effect of biochar on native soil microbial communities. Microbial analysis of environmental samples requires accurate and reproducible methods for the extraction of DNA from samples. Because of the variety among microbial species and the strong adsorption of the phosphate backbone of the DNA molecule to biochar, extracting and purifying high quality microbial DNA from biochar-amended soil is not a trivial process and can be considerably more difficult than the extraction of DNA from other environmental samples. The aim of this study was to compare the relative efficacies of three commercial DNA extraction kits, the FastDNA® SPIN Kit for Soil (FD kit), the PowerSoil® DNA Isolation Kit (PS kit) and the ZR Soil Microbe DNA Kit Miniprep™ (ZR kit), for extracting microbial genomic DNA from sand treated with different types of biochar. The methods were evaluated by comparing the DNA yields and purity and by analysing the bacterial and fungal community profiles generated by PCR-DGGE. Our results showed that the PCR-DGGE profiles for bacterial and fungal communities were highly affected by the purity and yield of the different DNA extracts. Among the tested kits, the PS kit was the most efficient with respect to the amount and purity of recovered DNA and considering the complexity of the generated DGGE microbial fingerprint from the sand-biochar samples. PMID:24948928

  4. Comparison of DNA extraction protocols for microbial communities from soil treated with biochar.

    PubMed

    Leite, D C A; Balieiro, F C; Pires, C A; Madari, B E; Rosado, A S; Coutinho, H L C; Peixoto, R S

    2014-01-01

    Many studies have evaluated the effects of biochar application on soil structure and plant growth. However, there are very few studies describing the effect of biochar on native soil microbial communities. Microbial analysis of environmental samples requires accurate and reproducible methods for the extraction of DNA from samples. Because of the variety among microbial species and the strong adsorption of the phosphate backbone of the DNA molecule to biochar, extracting and purifying high quality microbial DNA from biochar-amended soil is not a trivial process and can be considerably more difficult than the extraction of DNA from other environmental samples. The aim of this study was to compare the relative efficacies of three commercial DNA extraction kits, the FastDNA® SPIN Kit for Soil (FD kit), the PowerSoil® DNA Isolation Kit (PS kit) and the ZR Soil Microbe DNA Kit Miniprep™ (ZR kit), for extracting microbial genomic DNA from sand treated with different types of biochar. The methods were evaluated by comparing the DNA yields and purity and by analysing the bacterial and fungal community profiles generated by PCR-DGGE. Our results showed that the PCR-DGGE profiles for bacterial and fungal communities were highly affected by the purity and yield of the different DNA extracts. Among the tested kits, the PS kit was the most efficient with respect to the amount and purity of recovered DNA and considering the complexity of the generated DGGE microbial fingerprint from the sand-biochar samples.

  5. Role of polyadenylic acid in a deoxyribonucleic acid-membrane fraction extracted from pneumococci.

    PubMed Central

    Firshein, W; Meyer, B; Epner, E; Viggiani, J

    1976-01-01

    After the addition of radioactive polyadenylic acid to cell suspensions of pneumocci, part of the radioactivity becomes associated with a deoxyribonucleic acid (DNA)-membrane fraction extracted from the cells. A variety of techniques show that a portion of this associated radioactivity may represent oligoadenylates complexed to DNA, probaby as part of a ribonucleic acid (RNA) component. Polyadenylic acid, which had previously been shown to enhance DNA synthesis in cell suspensions (Firshein and Benson, 1968), also enhances the extent of DNA synthesis by the DNA-membrane fraction in vitro under specific conditions of concentration and conformation. The mechanism of action of this enhancement may be related to the ability of oligoadenylates to increase the number of initiation sites for DNA replication by stimulating the production of an RNA primer, thus providing additional 3'-OH groups with which DNA polymerase can react. PMID:6428

  6. Improved extraction of saturated fatty acids but not omega-3 fatty acids from sheep red blood cells using a one-step extraction procedure.

    PubMed

    Clayton, Edward H; Gulliver, Catherine E; Piltz, John W; Taylor, Robert D; Blake, Robert J; Meyer, Richard G

    2012-07-01

    Several methods are available to extract total lipid and methylate fatty acids from a range of samples including red blood cells (RBC). Fatty acid analysis of human RBC can be undertaken using a two-step extraction and methylation or a combined one-step extraction and methylation procedure. The lipid composition of sheep RBC differs significantly from that of humans and may affect their extraction. The purpose of the current study was to examine the efficiency of extraction of lipid and detection of fatty acids from sheep RBC using a one-step procedure. Fatty acids were analysed using a one-step extraction and methylation procedure using methanol:toluene and acetyl chloride in comparison with a two-step procedure involving extraction of lipid using chloroform:methanol and separate methylation. Concentrations of saturated fatty acids including C16:0 and C18:0 were significantly higher (42.6 and 33.9 % respectively) following extraction using the one-step procedure compared with the two-step procedure. However, concentrations of some polyunsaturated fatty acids, including C20:5n-3 and C22:6n-3 were not significantly different between either procedure. The improved detection of fatty acids may be related to the ability of different solvents to extract different lipid fractions. The differential extraction of lipids and detection of fatty acids from sheep RBC may have important implications in studies examining the effect of dietary treatment on the possible health benefits of fatty acids.

  7. A modified acidic approach for DNA extraction from plant species containing high levels of secondary metabolites.

    PubMed

    Cavallari, M M; Siqueira, M V B M; Val, T M; Pavanelli, J C; Monteiro, M; Grando, C; Pinheiro, J B; Zucchi, M I; Gimenes, M A

    2014-08-25

    Purified genomic DNA can be difficult to obtain from some plant species because of the presence of impurities such as polysaccharides, which are often co-extracted with DNA. In this study, we developed a fast, simple, and low-cost protocol for extracting DNA from plants containing high levels of secondary metabolites. This protocol does not require the use of volatile toxic reagents such as mercaptoethanol, chloroform, or phenol and allows the extraction of high-quality DNA from wild and cultivated tropical species.

  8. Comparative evaluation of commercially available manual and automated nucleic acid extraction methods for rotavirus RNA detection in stools.

    PubMed

    Esona, Mathew D; McDonald, Sharla; Kamili, Shifaq; Kerin, Tara; Gautam, Rashi; Bowen, Michael D

    2013-12-01

    Rotaviruses are a major cause of viral gastroenteritis in children. For accurate and sensitive detection of rotavirus RNA from stool samples by reverse transcription-polymerase chain reaction (RT-PCR), the extraction process must be robust. However, some extraction methods may not remove the strong RT-PCR inhibitors known to be present in stool samples. The objective of this study was to evaluate and compare the performance of six extraction methods used commonly for extraction of rotavirus RNA from stool, which have never been formally evaluated: the MagNA Pure Compact, KingFisher Flex and NucliSENS easyMAG instruments, the NucliSENS miniMAG semi-automated system, and two manual purification kits, the QIAamp Viral RNA kit and a modified RNaid kit. Using each method, total nucleic acid or RNA was extracted from eight rotavirus-positive stool samples with enzyme immunoassay optical density (EIA OD) values ranging from 0.176 to 3.098. Extracts prepared using the MagNA Pure Compact instrument yielded the most consistent results by qRT-PCR and conventional RT-PCR. When extracts prepared from a dilution series were extracted by the 6 methods and tested, rotavirus RNA was detected in all samples by qRT-PCR but by conventional RT-PCR testing, only the MagNA Pure Compact and KingFisher Flex extracts were positive in all cases. RT-PCR inhibitors were detected in extracts produced with the QIAamp Viral RNA Mini kit. The findings of this study should prove useful for selection of extraction methods to be incorporated into future rotavirus detection and genotyping protocols.

  9. Profiling of hydroxycinnamoylquinic acids in plant extracts using in-source CID fragmentation.

    PubMed

    Nagy, Ádám; Abrankó, László

    2016-12-01

    Hydroxycinnamoylquinic acids (HCQAs) are a major class of phenolic plant secondary metabolites, belonging to the chlorogenic acid family. Various health-beneficial properties of HCQAs have been shown, which has drawn interest for HCQA profiling in plants of human consumption. However, this task remains challenging, because several isomeric HCQAs can be present in the sample with identical molecular formulae and the limited availability of reference standards poses additional challenges to their identification. In the present work, a high performance liquid chromatography-electrospray ionization-quadrupole time-of-flight-mass spectrometry (HPLC-ESI-Q/TOF-MS) method accompanied with an effective data filtering protocol is presented, which is shown to be suitable for the identification of HCQAs in plant materials in a non-targeted manner. Both collision-induced dissociation (CID) fragmentation performed in a collision cell and in-source (CID) fragmentation were used to produce accurate mass fragments. It was shown that fragmentation characteristics required for identification of regio-isomers of HCQAs can be achieved with in-source CID fragmentation, enabling the use of a single-stage MS system with in-source fragmentation for convincing identification of HCQAs. Based on a thorough validation of identified HCQA compounds using coffee bean extracts as reference samples, comprehensive profiling of HCQAs in two apricot (Prunus armeniaca L.) genotypes ('Preventa' and 'Gönci magyarkajszi') was carried out for the first time and the following 10 HCQAs were shown to be present in apricot fruit: 3-caffeoylquinic acid (CQA), cis-3-CQA, 4-CQA, 5-CQA, cis-5-CQA, 3,5-diCQA, 3-p-coumaroylquinic acid (pCoQA), 4-pCoQA, 3-feruloylquinic acid (FQA) and cis-3-FQA. Copyright © 2016 John Wiley & Sons, Ltd.

  10. Antioxidant activity and sensory assessment of a rosmarinic acid-enriched extract of Salvia officinalis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An extract of S. officinalis (garden sage) was prepared using supercritical carbon dioxide (SC-CO2) extraction, followed by a Soxhlet hot water extraction. The resulting extract was enriched in polyphenols, including rosmarinic acid (RA), which has shown promising health benefits in animals. The ext...

  11. Optimization of an extraction protocol for organic matter from soils and sediments using high resolution mass spectrometry: selectivity and biases

    NASA Astrophysics Data System (ADS)

    Chu, R. K.; Tfaily, M. M.; Tolic, N.; Kyle, J. E.; Robinson, E. R.; Hess, N. J.; Paša-Tolić, L.

    2015-12-01

    Soil organic matter (SOM) is a complex mixture of above and belowground plant litter and microbial residues, and is a key reservoir for carbon (C) and nutrient biogeochemical cycling in different ecosystems. A limited understanding of the molecular composition of SOM prohibits the ability to routinely decipher chemical processes within soil and predict how terrestrial C fluxes will response to changing climatic conditions. Here, we present that the choice of solvent can be used to selectively extract different compositional fractions from SOM to either target a specific class of compounds or gain a better understanding of the entire composition of the soil sample using 12T Fourier transform ion cyclotron resonance mass spectrometry. Specifically, we found that hexane and chloroform were selective for lipid-like compounds with very low O:C ratios; water was selective for carbohydrates with high O:C ratios; acetonitrile preferentially extracts lignin, condensed structures, and tannin polyphenolic compounds with O:C > 0.5; methanol has higher selectivity towards lignin and lipid compounds characterized with relatively low O:C < 0.5. Hexane, chloroform, methanol, acetonitrile and water increase the number and types of organic molecules extracted from soil for a broader range of chemically diverse soil types. Since each solvent extracts a selective group of compounds, using a suite of solvents with varying polarity for analysis results in more comprehensive representation of the diversity of organic molecules present in soil and a better representation of the whole spectrum of available substrates for microorganisms. Moreover, we have developed a sequential extraction protocol that permits sampling diverse classes of organic compounds while minimizing ionization competition during ESI while increasing sample throughput and decreasing sample volume. This allowed us to hypothesize about possible chemical reactions relating classes of organic molecules that reflect abiotic

  12. Extraction equilibrium of indium(III) from nitric acid solutions by di(2-ethylhexyl)phosphoric acid dissolved in kerosene.

    PubMed

    Tsai, Hung-Sheng; Tsai, Teh-Hua

    2012-01-04

    The extraction equilibrium of indium(III) from a nitric acid solution using di(2-ethylhexyl) phosphoric acid (D2EHPA) as an acidic extractant of organophosphorus compounds dissolved in kerosene was studied. By graphical and numerical analysis, the compositions of indium-D2EHPA complexes in organic phase and stoichiometry of the extraction reaction were examined. Nitric acid solutions with various indium concentrations at 25 °C were used to obtain the equilibrium constant of InR₃ in the organic phase. The experimental results showed that the extraction distribution ratios of indium(III) between the organic phase and the aqueous solution increased when either the pH value of the aqueous solution and/or the concentration of the organic phase extractant increased. Finally, the recovery efficiency of indium(III) in nitric acid was measured.

  13. A rational protocol for the successful crystallization of l-amino-acid oxidase from Bothrops atrox

    PubMed Central

    Alves, Raquel Melo; Feliciano, Patricia Rosa; Sampaio, Suely Vilela; Nonato, Maria Cristina

    2011-01-01

    Despite the valuable contributions of robotics and high-throughput approaches to protein crystallization, the role of an experienced crystallographer in the evaluation and rationalization of a crystallization process is still crucial to obtaining crystals suitable for X-ray diffraction measurements. In this work, the difficult task of crystallizing the flavoenzyme l-amino-acid oxidase purified from Bothrops atrox snake venom was overcome by the development of a protocol that first required the identification of a non-amorphous precipitate as a promising crystallization condition followed by the implementation of a methodology that combined crystallization in the presence of oil and seeding techniques. Crystals were obtained and a complete data set was collected to 2.3 Å resolution. The crystals belonged to space group P21, with unit-cell parameters a = 73.64, b = 123.92, c = 105.08 Å, β = 96.03°. There were four protein subunits in the asymmetric unit, which gave a Matthews coefficient V M of 2.12 Å3 Da−1, corresponding to 42% solvent content. The structure has been solved by molecular-replacement techniques. PMID:21505245

  14. Detection efficiency and noise in a semi-device-independent randomness-extraction protocol

    NASA Astrophysics Data System (ADS)

    Li, Hong-Wei; Yin, Zhen-Qiang; Pawłowski, Marcin; Guo, Guang-Can; Han, Zheng-Fu

    2015-03-01

    In this paper, we analyze several critical issues in semi-device-independent quantum information processing protocol. In practical experimental realization, randomness generation in that scenario is possible only if the efficiency of the detectors used is above a certain threshold. Our analysis shows that the critical detection efficiency is √{2/}2 in the symmetric setup, while in the asymmetric setup if one of the bases has perfect critical detection efficiency then the other one can be arbitrarily close to 0. We also analyze the semi-device-independent random-number-generation efficiency based on different averages of guessing probability. To generate more randomness, the proper averaging method should be applied. Its choice depends on the value of a certain dimension witness. More importantly, the general analytical relationship between the maximal average guessing probability and dimension witness is given.

  15. Application of a high-level peracetic acid disinfection protocol to re-process antibiotic disinfected skin allografts.

    PubMed

    Lomas, R J; Huang, Q; Pegg, D E; Kearney, J N

    2004-01-01

    Skin allografts, derived from cadaveric donors, are widely used for the treatment of burns and ulcers. Prior to use in clinical situations, these allografts are disinfected using a cocktail of antibiotics and then cryopreserved. Unfortunately, this antibiotic disinfection procedure fails to decontaminate a significant proportion and these contaminated grafts can not be used clinically. We have investigated whether it is possible to apply a second, more potent disinfection procedure to these contaminated grafts and effectively to re-process them for clinical use. Cadaveric skin grafts, treated with antibiotics and cryopreserved, were thawed and a peracetic acid (PAA) disinfection protocol applied. The grafts were then preserved in a high concentration of glycerol or propylene glycol, and properties thought to be essential for successful clinical performance assessed. The cytotoxicity of the grafts was assessed using both extract and contact assays; damage to the skin collagen was assessed using a collagenase susceptibility assay and the capacity of the grafts to elicit an inflammatory response in vitro was assessed by quantifying the production of the pro-inflammatory cytokine TNF-alpha by human peripheral blood mononuclear phagocytes. PAA disinfection, in conjunction with either glycerol or propylene glycol preservation, did not render the grafts cytotoxic, pro-inflammatory, or increase their susceptibility to collagenase digestion. The rates of penetration of glycerol and propylene glycol into the re-processed skin were comparable to those of fresh skin. This study has demonstrated that PAA disinfection combined with immersion in high concentrations of either glycerol or propylene glycol was an effective method for re-processing contaminated skin allografts, and may justify their clinical use.

  16. Microbes on building materials--evaluation of DNA extraction protocols as common basis for molecular analysis.

    PubMed

    Ettenauer, Jörg D; Piñar, Guadalupe; Lopandic, Ksenija; Spangl, Bernhard; Ellersdorfer, Günther; Voitl, Christian; Sterflinger, Katja

    2012-11-15

    The study of microbial life in building materials is an emerging topic concerning biodeterioration of materials as well as health risks in houses and at working places. Biodegradation and potential health implications associated with microbial growth in our residues claim for more precise methods for quantification and identification. To date, cultivation experiments are commonly used to gain insight into the microbial diversity. Nowadays, molecular techniques for the identification of microorganisms provide efficient methods that can be applied in this field. The efficiency of DNA extraction is decisive in order to perform a reliable and reproducible quantification of the microorganisms by qPCR or to characterize the structure of the microbial community. In this study we tested thirteen DNA extraction methods and evaluated their efficiency for identifying (1) the quantity of DNA, (2) the quality and purity of DNA and (3) the ability of the DNA to be amplified in a PCR reaction using three universal primer sets for the ITS region of fungi as well as one primer pair targeting the 16S rRNA of bacteria with three typical building materials - common plaster, red brick and gypsum cardboard. DNA concentration measurements showed strong variations among the tested methods and materials. Measurement of the DNA yield showed up to three orders of magnitude variation from the same samples, whereas A260/A280 ratios often prognosticated biases in the PCR amplifications. Visualization of the crude DNA extracts and the comparison of DGGE fingerprints showed additional drawbacks of some methods. The FastDNA Spin kit for soil showed to be the best DNA extraction method and could provide positive results for all tests with the three building materials. Therefore, we suggest this method as a gold standard for quantification of indoor fungi and bacteria in building materials.

  17. Novel Regenerated Solvent Extraction Processes for the Recovery of Carboxylic Acids or Ammonia from Aqueous Solutions Part I. Regeneration of Amine-Carboxylic Acid Extracts

    SciTech Connect

    Poole, L.J.; King, C.J.

    1990-03-01

    Two novel regenerated solvent extraction processes are examined. The first process has the potential to reduce the energy costs inherent in the recovery of low-volatility carboxylic acids from dilute aqueous solutions. The second process has the potential for reducing the energy costs required for separate recovery of ammonia and acid gases (e.g. CO{sub 2} and H{sub 2}S) from industrial sour waters. The recovery of carboxylic acids from dilute aqueous solution can be achieved by extraction with tertiary amines. An approach for regeneration and product recovery from such extracts is to back-extract the carboxylic acid with a water-soluble, volatile tertiary amine, such as trimethylamine. The resulting trimethylammonium carboxylate solution can be concentrated and thermally decomposed, yielding the product acid and the volatile amine for recycle. Experimental work was performed with lactic acid, succinic acid, and fumaric acid. Equilibrium data show near-stoichiometric recovery of the carboxylic acids from an organic solution of Alamine 336 into aqueous solutions of trimethylamine. For fumaric and succinic acids, partial evaporation of the aqueous back extract decomposes the carboxylate and yields the acid product in crystalline form. The decomposition of aqueous solutions of trimethylammonium lactates was not carried out to completion, due to the high water solubility of lactic acid and the tendency of the acid to self-associate. The separate recovery of ammonia and acid gases from sour waters can be achieved by combining steam-stripping of the acid gases with simultaneous removal of ammonia by extraction with a liquid cation exchanger. The use of di-2,4,4-trimethylpentyl phosphinic acid as the liquid cation exchanger is explored in this work. Batch extraction experiments were carried out to measure the equilibrium distribution ratio of ammonia between an aqueous buffer solution and an organic solution of the phosphinic acid (0.2N) in Norpar 12. The concentration

  18. Extraction of amino acids from soils and sediments with superheated water

    NASA Technical Reports Server (NTRS)

    Cheng, C. N.; Ponnamperuma, C.

    1974-01-01

    A method of extraction for amino acids from soils and sediments involving superheated water has been investigated. About 75-97 per cent of the amino acids contained in four soils of a soil profile from Illinois were extracted by this method. Deep penetration of water into soil aggregates and partial hydrolysis of peptide bonds during this extraction by water at high temperature are likely mechanisms responsible for the release of amino acids from samples. This extraction method does not require subsequent desalting treatments when analyses are carried out with an ion-exchange amino acid analyzer.

  19. Full-length protein extraction protocols and gel-based downstream applications in formalin-fixed tissue proteomics.

    PubMed

    Tanca, Alessandro; Uzzau, Sergio; Addis, Maria Filippa

    2015-01-01

    Archival formalin-fixed, paraffin-embedded (FFPE) tissue repositories and their associated clinical information can represent a valuable resource for tissue proteomics. In order to make these tissues available for protein biomarker discovery and validation studies, dedicated sample preparation procedures overcoming the intermolecular cross-links introduced by formalin need to be implemented. This chapter describes a full-length protein extraction protocol optimized for downstream gel-based proteomics applications. Using the procedures detailed here, SDS-PAGE, western immunoblotting, GeLC-MS/MS, 2D-PAGE, and 2D-DIGE can be carried out on FFPE tissues. Technical tips, critical aspects, and drawbacks of the method are presented and discussed.

  20. Influence of the protocol used for fibroin extraction on the mechanical properties and fiber sizes of electrospun silk mats.

    PubMed

    Aznar-Cervantes, Salvador D; Vicente-Cervantes, Daniel; Meseguer-Olmo, Luis; Cenis, José L; Lozano-Pérez, A Abel

    2013-05-01

    Silk fibroin (SF) was regenerated using three of the most common protocols described in the bibliography for the dissolution of raw SF (LiBr 9.3M, CaCl2 50 wt.% or CaCl2:EtOH:H2O 1:2:8 in molar ratio). The integrity of regenerated SF in aqueous solution was analyzed by SDS-PAGE and different profiles of degradation were observed depending on the protocol used. This fact was found to affect also the aqueous solubility of the freeze dried protein. These different SFs were used to produce electrospun mats using SF solutions of SF 17 wt.% in 1,1,1,1',1',1'-hexafluoro-2-propanol (HFIP) and significant differences in fiber sizes, elongation and ultimate strength values were found. This work provides a global overview of the manner that different methods of SF extraction can affect the properties of electrospun SF-mats and consequently it should be considered depending on the use they are going to be made for.

  1. Method for extracting lanthanides and actinides from acid solutions by modification of Purex solvent

    DOEpatents

    Horwitz, E.P.; Kalina, D.G.

    1984-05-21

    A process has been developed for the extraction of multivalent lanthanide and actinide values from acidic waste solutions, and for the separation of these values from fission product and other values, which utilizes a new series of neutral bi-functional extractants, the alkyl(phenyl)-N, N-dialkylcarbamoylmethylphosphine oxides, in combination with a phase modifier to form an extraction solution. The addition of the extractant to the Purex process extractant, tri-n-butylphosphate in normal paraffin hydrocarbon diluent, will permit the extraction of multivalent lanthanide and actinide values from 0.1 to 12.0 molar acid solutions.

  2. Choice of solvent extraction technique affects fatty acid composition of pistachio (Pistacia vera L.) oil.

    PubMed

    Abdolshahi, Anna; Majd, Mojtaba Heydari; Rad, Javad Sharifi; Taheri, Mehrdad; Shabani, Aliakbar; Teixeira da Silva, Jaime A

    2015-04-01

    Pistachio (Pistacia vera L.) oil has important nutritional and therapeutic properties because of its high concentration of essential fatty acids. The extraction method used to obtain natural compounds from raw material is critical for product quality, in particular to protect nutritional value. This study compared the fatty acid composition of pistachio oil extracted by two conventional procedures, Soxhlet extraction and maceration, analyzed by a gas chromatography-flame ionization detector (GC-FID). Four solvents with different polarities were tested: n-hexane (Hx), dichloromethane (DCM), ethyl acetate (EtAc) and ethanol (EtOH). The highest unsaturated fatty acid content (88.493 %) was obtained by Soxhlet extraction with EtAc. The Soxhlet method extracted the most oleic and linolenic acids (51.99 % and 0.385 %, respectively) although a higher concentration (36.32 %) of linoleic acid was extracted by maceration.

  3. Characterization of citrus pectin samples extracted under different conditions: influence of acid type and pH of extraction

    PubMed Central

    Kaya, Merve; Sousa, António G.; Crépeau, Marie-Jeanne; Sørensen, Susanne O.; Ralet, Marie-Christine

    2014-01-01

    Background and Aims Pectin is a complex macromolecule, the fine structure of which is influenced by many factors. It is used as a gelling, thickening and emulsifying agent in a wide range of applications, from food to pharmaceutical products. Current industrial pectin extraction processes are based on fruit peel, a waste product from the juicing industry, in which thousands of tons of citrus are processed worldwide every year. This study examines how pectin components vary in relation to the plant source (orange, lemon, lime, grapefruit) and considers the influence of extraction conditions on the chemical and macromolecular characteristics of pectin samples. Methods Citrus peel (orange, lemon, lime and grapefruit) from a commercial supplier was used as raw material. Pectin samples were obtained on a bulk plant scale (kilograms; harsh nitric acid, mild nitric acid and harsh oxalic acid extraction) and on a laboratory scale (grams; mild oxalic acid extraction). Pectin composition (acidic and neutral sugars) and physicochemical properties (molar mass and intrinsic viscosity) were determined. Key Results Oxalic acid extraction allowed the recovery of pectin samples of high molecular weight. Mild oxalic acid-extracted pectins were rich in long homogalacturonan stretches and contained rhamnogalacturonan I stretches with conserved side chains. Nitric acid-extracted pectins exhibited lower molecular weights and contained rhamnogalacturonan I stretches encompassing few and/or short side chains. Grapefruit pectin was found to have short side chains compared with orange, lime and lemon. Orange and grapefruit pectin samples were both particularly rich in rhamnogalacturonan I backbones. Conclusions Structural, and hence macromolecular, variations within the different citrus pectin samples were mainly related to their rhamnogalacturonan I contents and integrity, and, to a lesser extent, to the length of their homogalacturonan domains. PMID:25081519

  4. Determination of free medium-chain fatty acids in beer by stir bar sorptive extraction.

    PubMed

    Horák, Tomás; Culík, Jirí; Jurková, Marie; Cejka, Pavel; Kellner, Vladimír

    2008-07-04

    Free medium-chain fatty acids in beer originate from raw materials, mainly from the fermentation activity of yeasts, and can influence beer taste, vitality of yeasts and also the foam stability of beer. This study presents the development of the method for the determination of free fatty medium-chain acids including caproic acid, caprylic acid, capric acid and lauric acid in beer or wort using stir bar sorptive extraction (SBSE). The combination of this extraction technique with solvent back extraction of the extracted analytes and subsequent gas chromatographic analysis with flame ionization detection was used for the determination of these compounds. The influences of different solvent back solutions, sampling time, solvent back extraction times and different contents of ethanol were studied. The method had high repeatability (RSD <6.7%), good linearity (the correlation coefficients were higher than 0.9963 for quadratic curves over the concentration range 0.5-8.0mg/l) and recoveries 57-89%.

  5. Optimization of the derivatization protocol of pentacyclic triterpenes prior to their gas chromatography-mass spectrometry analysis in plant extracts.

    PubMed

    Jemmali, Zaineb; Chartier, Agnes; Dufresne, Christelle; Elfakir, Claire

    2016-01-15

    This paper focuses on the application of a two-level full factorial design to optimize the key derivatization step before the GC-FID and GC-MS analysis of pentacyclic triterpenes. The derivatization reaction was screened for influential factors and statistically significant parameters with a p value less than 0.05. A multi-response optimization based on a desirability function was then applied, while simultaneously considering overall detection enhancement of compounds. Results showed that derivatization using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) and trimethylchlorosilane (TMCS) in pyridine (22:13:65v/v/v) for 2h at 30°C was the most efficient method of derivatizing all the hydroxyl and carboxylic acid groups contained in the triterpene structures. The validity of the method was demonstrated using GC-MS analyzes of a mixture containing eleven standards (β-amyrin, α-amyrin, lupeol, erythrodiol, uvaol, betulin, oleanolic acid, betulinic acid, ursolic acid, maslinic acid and corosolic acid). These compounds are representative of different classes of terpene compounds bearing different functional groups such as alcohols, diols, and carboxylic acids. The derivatization procedure was then tested on four plant extracts: apple pomace, salvia sclarea (dried leaves and flowers), sea buckthorn (Hyppophae rhammnoides L.) berries, and B. serrata resin. The identification of triterpenes was based on the comparison of their retention time and mass spectra to those of standards. The presence of compounds already identified in the literature was confirmed and new ones such as maslinic and corosolic acids were identified in apples, sea buckthorn and salvia sclarea.

  6. Synthesis of caffeic acid and p-hydroxybenzoic acid molecularly imprinted polymers and their application for the selective extraction of polyphenols from olive mill waste waters.

    PubMed

    Michailof, Chrysa; Manesiotis, Panagiotis; Panayiotou, Costas

    2008-02-22

    Using caffeic acid and p-hydroxybenzoic acid as templates, two molecularly imprinted polymers (MIPs) were prepared that were used for isolation of polyphenols from olive mill waste water samples (OMWWs) without previous pre-treatment. For the preparation of the caffeic acid MIPs 4-vinylpyridine, allylurea, allylaniline and methacrylic acid were tested as functional monomers, ethylene glycol dimethylacrylate (EDMA), pentaerythritol trimethylacrylate (PETRA) and divinylbenzene 80 (DVB80) as cross-linkers and tetrahydrofuran as porogen. For p-hydroxybenzoic acid 4-vinylpyridine, allylurea and allylaniline were tested as functional monomers, EDMA and PETRA as cross-linkers and acetonitrile as porogen. The performance of the synthesized polymers was evaluated against seven structurally related compounds by means of polymer-based HPLC. The two polymers that presented the most interesting properties were further evaluated by batch rebinding and from the derived isotherms their capacity and binding strength were determined. Using solid-phase extraction (SPE), their ability to recognize and bind the template molecule from an aqueous solution as well as the pH dependence of the binding strength were explored. After establishing the best SPE protocol, an aqueous model mixture of compounds and a raw OMWWs sample were loaded on the two best polymers. The result of the consecutive use of the two polymers on the same sample was explored. It was concluded that acidic conditions favour the recognition abilities of both polymers and that they can be used for a quick and efficient isolation of the polyphenol fraction directly from raw OMWW.

  7. Antioxidant activity and sensory evaluation of a rosmarinic acid-enriched extract of Salvia officinalis.

    PubMed

    Bakota, Erica L; Winkler-Moser, Jill K; Berhow, Mark A; Eller, Fred J; Vaughn, Steven F

    2015-04-01

    An extract of Salvia officinalis (garden sage) was prepared using supercritical carbon dioxide (SC-CO2 ) extraction, followed by hot water extraction. The resulting extract was enriched in polyphenols, including rosmarinic acid (RA), which has shown promising health benefits in animals. The extract contained RA at a concentration of 28.4 mg/g, representing a significant enrichment from the RA content in sage leaves. This extract was incorporated into oil-in-water emulsions as a source of lipid antioxidants and compared to emulsions containing pure rosmarinic acid. Both treatments were effective in suppressing lipid oxidation. The extract was evaluated by a trained sensory panel in a tea formulation. While the panel could discriminate among extract-treated and control samples, panelists demonstrated high acceptability of the sage extract in a tea.

  8. 21 CFR 573.500 - Condensed, extracted glutamic acid fermentation product.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.500 Condensed, extracted glutamic acid... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Condensed, extracted glutamic acid...

  9. 21 CFR 573.500 - Condensed, extracted glutamic acid fermentation product.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.500 Condensed, extracted glutamic acid... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Condensed, extracted glutamic acid...

  10. 21 CFR 573.500 - Condensed, extracted glutamic acid fermentation product.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.500 Condensed, extracted glutamic acid... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Condensed, extracted glutamic acid...

  11. 21 CFR 573.500 - Condensed, extracted glutamic acid fermentation product.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.500 Condensed, extracted glutamic acid... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Condensed, extracted glutamic acid...

  12. The behavior and importance of lactic acid complexation in Talspeak extraction systems

    SciTech Connect

    Grimes, Travis S.; Nilsson, Mikael; Nash, Kenneth L.

    2008-07-01

    Advanced partitioning of spent nuclear fuel in the UREX +la process relies on the TALSPEAK process for separation of fission-product lanthanides from trivalent actinides. The classic TALSPEAK utilizes an aqueous medium of both lactic acid and diethylenetriaminepentaacetic acid and the extraction reagent di(2-ethylhexyl)phosphoric acid in an aromatic diluent. In this study, the specific role of lactic acid and the complexes involved in the extraction of the trivalent actinides and lanthanides have been investigated using {sup 14}C-labeled lactic acid. Our results show that lactic acid partitions between the phases in a complex fashion. (authors)

  13. Solvent extraction in the treatment of acidic high-level liquid waste : where do we stand?

    SciTech Connect

    Horwitz, E. P.; Schulz, W. W.

    1998-06-18

    During the last 15 years, a number of solvent extraction/recovery processes have been developed for the removal of the transuranic elements, {sup 90}Sr and {sup 137}Cs from acidic high-level liquid waste. These processes are based on the use of a variety of both acidic and neutral extractants. This chapter will present an overview and analysis of the various extractants and flowsheets developed to treat acidic high-level liquid waste streams. The advantages and disadvantages of each extractant along with comparisons of the individual systems are discussed.

  14. Molecularly imprinted polymer microspheres for solid-phase extraction of protocatechuic acid in Rhizoma homalomenae.

    PubMed

    Chen, Fang-Fang; Wang, Guo-Ying; Shi, Yan-Ping

    2011-10-01

    Molecularly imprinted polymers (MIPs) had been prepared by precipitation polymerization method using acrylamide as the functional monomer, ethylene glycol dimethacrylate as the cross-linker, acetonitrile as the porogen solvent and protocatechuic acid (PA), one of phenolic acids, as the template molecule. The MIPs were characterized by scanning electron microscopy and Fourier transform infrared, and their performance relative to non-imprinted polymers was assessed by equilibrium binding experiments. Six structurally similar phenolic acids, including p-hydroxybenzoic acid, gallic acid, salicylic acid, syringic acid, vanillic acid, ferulic acid were selected to assess the selectivity and recognition capability of the MIPs. The MIPs were applied to extract PA from the traditional Chinese medicines as a solid-phase extraction sorbent. The resultant cartridge showed that the MIPs have a good extraction performance and were able to selectively extract almost 82% of PA from the extract of Rhizoma homalomenae. Thus, the proposed molecularly imprinted-solid phase extraction-high performance liquid chromatography method can be successfully used to extract and analyse PA in traditional Chinese medicines.

  15. Determination of fatty acid composition and quality characteristics of oils from palm fruits using solvent extraction

    NASA Astrophysics Data System (ADS)

    Kasmin, Hasimah; Lazim, Azwan Mat; Awang, Roila

    2015-09-01

    Palm oil contains about 45% of saturated palmitic acid and 39% of mono-unsaturated oleic acid. Investigations made in the past to trace the fatty acid composition in palm revealed that ripeness of fresh fruit bunch (FFB) affect oil composition. However, there is no evidence that processing operations affect oil composition, although different stage of processing does affect the quality of oil extracted. An improved method for sterilizing the oil palm fruits by dry heating, followed by oil extraction has been studied. This method eliminates the use of water, thus, increasing the extraction of lipid soluble. The objective of this study is to determine the possibility production of palm oil with different fatty acid composition (FAC) as well as the changes in quality from conventional milling. The unripe and ripe FFB were collected, sterilized and extracted using different method of solvent extraction. Preliminary data have shown that variation in FAC will also alter the physical and chemical properties of the oil extracted.

  16. The leaf extract of Siberian Crabapple (Malus baccata (Linn.) Borkh) contains potential fatty acid synthase inhibitors.

    PubMed

    Wei, Xiang; Zhao, Ran; Sun, Ying-Hui; Cong, Jian-Ping; Meng, Fan-Guo; Zhou, Hai-Meng

    2009-02-01

    The present work focused on the kinetics of the inhibitory effects of the leaf extract of Siberian Crabapple, named Shan jingzi in China, on chicken liver fatty acid synthase. The results showed that this extract had much stronger inhibitory ability on fatty acid synthase than that from green teas described in many previous reports. The inhibitory ability of this extract is closely related to the extracting solvent, and the time of extraction was also an important influencing factor. The inhibitory types of this extract on diffeerent substrates of chicken liver fatty acid synthase, acetyl-CoA, malonyl-CoA and NADPH, were found to be noncompetitive, uncompetitive and mixed, respectively. The studies here shed a new light on the exploration for inhibitors of fatty acid synthase.

  17. Comparison of nucleic acid extraction methods for the detection of Mycoplasma pneumoniae.

    PubMed

    Thurman, Kathleen A; Cowart, Kelley C; Winchell, Jonas M

    2009-12-01

    Four nucleic acid extraction procedures (2 automated and 2 manual) were compared for their efficiency at isolating Mycoplasma pneumoniae DNA. Oropharyngeal swabs from healthy volunteers were spiked with varying amounts of M. pneumoniae, extracted, and tested using real-time polymerase chain reaction. Our data indicate that both automated extraction methods consistently outperform the manual procedures.

  18. DEVELOPMENTS IN THE SUPERCRITICAL FLUID EXTRACTION OF CHLOROPHENOXY ACID HERBICIDES FROM SOIL SAMPLES

    EPA Science Inventory

    Extraction of chlorophenoxy acid herbicides from soil samples with supercritical carbon dioxide as extractant and tetrabutylammonium hydroxide and methyl iodide as derivatization agents was investigated. The extraction was carried out at 400 atm and 80 C for 15 min static, follow...

  19. Discovery of novel, non-acidic mPGES-1 inhibitors by virtual screening with a multistep protocol

    PubMed Central

    Noha, Stefan M.; Fischer, Katrin; Koeberle, Andreas; Garscha, Ulrike; Werz, Oliver; Schuster, Daniela

    2015-01-01

    Microsomal prostaglandin E2 synthase-1 (mPGES-1) inhibitors are considered as potential therapeutic agents for the treatment of inflammatory pain and certain types of cancer. So far, several series of acidic as well as non-acidic inhibitors of mPGES-1 have been discovered. Acidic inhibitors, however, may have issues, such as loss of potency in human whole blood and in vivo, stressing the importance of the design and identification of novel, non-acidic chemical scaffolds of mPGES-1 inhibitors. Using a multistep virtual screening protocol, the Vitas-M compound library (∼1.3 million entries) was filtered and 16 predicted compounds were experimentally evaluated in a biological assay in vitro. This approach yielded two molecules active in the low micromolar range (IC50 values: 4.5 and 3.8 μM, respectively). PMID:26088337

  20. Safety and efficacy of tranexamic acid in bleeding paediatric trauma patients: a systematic review protocol

    PubMed Central

    Urban, Denisa; Dehaeck, Ruben; Lorenzetti, Diane; Guilfoyle, Jonathan; Poon, Man-Chiu; Steele, MacGregor; Lardner, David; Ma, Irene Wai Yan; Brindle, Mary Elizabeth

    2016-01-01

    Introduction Trauma is the leading cause of death among children aged 1–18. Studies indicate that better control of bleeding could potentially prevent 10–20% of trauma-related deaths. The antifibrinolytic agent tranexamic acid (TxA) has shown promise in haemorrhage control in adult trauma patients. However, information on the potential benefits of TxA in children remains sparse. This review proposes to evaluate the current uses, benefits and adverse effects of TxA in the bleeding paediatric trauma population. Methods and analysis A structured search of bibliographic databases (eg, MEDLINE, EMBASE, PubMed, CINAHL, Cochrane CENTRAL) has been undertaken to retrieve randomised controlled trials and cohort studies that describe the use of TxA in paediatric trauma patients. To ensure that all relevant data were captured, the search did not contain any restrictions on language or publication time. After deduplication, citations will be screened independently by 2 authors, and selected for inclusion based on prespecified criteria. Data extraction and risk of bias assessment will be performed independently and in duplicate. Meta-analytic methods will be employed wherever appropriate. Ethics and dissemination This study will not involve primary data collection, and formal ethical approval will therefore not be required. The findings of this study will be disseminated through a peer-reviewed publication and at relevant conference meetings. Trial registration number CRD42016038023. PMID:27660323

  1. A comparison of DNA extraction protocols from blood spotted on FTA cards for the detection of tick-borne pathogens by Reverse Line Blot hybridization.

    PubMed

    Hailemariam, Zerihun; Ahmed, Jabbar Sabir; Clausen, Peter-Henning; Nijhof, Ard Menzo

    2017-01-01

    An essential step in the molecular detection of tick-borne pathogens (TBPs) in blood is the extraction of DNA. When cooled storage of blood under field conditions prior to DNA extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. We evaluated six DNA extraction methods from blood spotted on FTA Classic(®) cards (FTA cards), to determine the optimal protocol for the subsequent molecular detection of TBPs by PCR and the Reverse Line Blot hybridization assay (RLB). Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. DNA was also isolated from whole blood dilutions using a commercial kit. PCR/RLB results showed that washing of 3mm discs punched from FTA cards with FTA purification reagent followed by DNA extraction using Chelex(®) resin was the most sensitive procedure. The detection limit could be improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3mm discs proved to be most practical. The presented best practice method for the extraction of DNA from blood spotted on FTA cards will facilitate epidemiological studies on TBPs. It may be particularly useful for field studies where a cold chain is absent.

  2. A simple and rapid protocol of crude DNA extraction from apple trees for PCR and real-time PCR detection of 'Candidatus Phytoplasma mali'.

    PubMed

    Aldaghi, M; Massart, S; Dutrecq, O; Bertaccini, A; Jijakli, M H; Lepoivre, P

    2009-03-01

    Different PCR protocols have been established for detection of European fruit trees phytoplasmas; however the majority of the procedures for extracting phytoplasma DNA are complex, time consuming, and expensive, with a risk of contamination or loss of target DNA. In present study, a crude extract preparation method previously used to detect other plant pathogens was adapted to samples from apple trees infected by 'Candidatus Phytoplasma mali'. End-point and real-time PCR detection of 'Ca. P. mali' were used to compare this extraction procedure with an established method for efficient extraction of purified DNA. The crude extract proved fully adequate for phytoplasma detection in samples from 86 in vitro and 35 in vivo apple shoots or plants and 10 periwinkle plants. High inter- and intra-run reproducibility was obtained for phytoplasma detection with different TaqMan MGB- or SYBR Green-based real-time PCR protocols applied to the crude extracts. Real-time PCR applied to serially diluted crude and purified extracts revealed the same phytoplasma detection limit (dilution up to 10(5)). All results confirm the suitability of this simple, quick, efficient extraction technique for accurate detection of 'Ca. P. mali' in different types of apple and periwinkle samples.

  3. Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308

    PubMed Central

    Matrone, M.; Keid, L.B.; Rocha, V.C.M.; Vejarano, M.P.; Ikuta, C.Y.; Rodriguez, C.A.R.; Ferreira, F.; Dias, R.A.; Ferreira Neto, J.S

    2009-01-01

    The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04) or bronchial lymph nodes (p=0.004) and equal to the spleens (p=0.18). From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (p< 0.0001) and GT (p=0.0004). There was no difference between the PK and GT protocols (p=0.5). Some positive samples from the classical bacteriology were negative to the PCR and vice-versa. Therefore, the best strategy for B. abortus detection in the organs of aborted fetuses or calves born from infected cows is the use, in parallel, of isolation by classical bacteriology and the PCR, with the DNA extraction performed by the Boom protocol. PMID:24031391

  4. Antioxidant activities of rosemary (Rosmarinus Officinalis L.) extract, blackseed (Nigella sativa L.) essential oil, carnosic acid, rosmarinic acid and sesamol.

    PubMed

    Erkan, Naciye; Ayranci, Guler; Ayranci, Erol

    2008-09-01

    Antioxidant activities of three pure compounds: carnosic acid, rosmarinic acid and sesamol, as well as two plant extracts: rosemary extract and blackseed essential oil, were examined by applying DPPH and ABTS(+) radical-scavenging assays and the ferric thiocyanate test. All three test methods proved that rosemary extract had a higher antioxidant activity than blackseed essential oil. The order of antioxidant activity of pure compounds showed variations in different tests. This was attributed to structural factors of individual compounds. Phenolic contents of blackseed essential oil and rosemary extract were also determined. Rosemary extract was found to have a higher phenolic content than blackseed essential oil. This fact was utilised in explaining the higher antioxidant activity of rosemary extract.

  5. Antimicrobial activity of acid-hydrolyzed Citrus unshiu peel extract in milk.

    PubMed

    Min, Keun Young; Kim, Hyun Jung; Lee, Kyoung Ah; Kim, Kee-Tae; Paik, Hyun-Dong

    2014-01-01

    Citrus fruit (Citrus unshiu) peels were extracted with hot water and then acid-hydrolyzed using hydrochloric acid. Antimicrobial activities of acid-hydrolyzed Citrus unshiu peel extract were evaluated against pathogenic bacteria, including Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes. Antilisterial effect was also determined by adding extracts at 1, 2, and 4% to whole, low-fat, and skim milk. The cell numbers of B. cereus, Staph. aureus, and L. monocytogenes cultures treated with acid-hydrolyzed extract for 12h at 35°C were reduced from about 8log cfu/mL to <1log cfu/mL. Bacillus cereus was more sensitive to acid-hydrolyzed Citrus unshiu peel extract than were the other bacteria. The addition of 4% acid-hydrolyzed Citrus unshiu extracts to all types of milk inhibited the growth of L. monocytogenes within 1d of storage at 4°C. The results indicated that Citrus unshiu peel extracts, after acid hydrolysis, effectively inhibited the growth of pathogenic bacteria. These findings indicate that acid hydrolysis of Citrus unshiu peel facilitates its use as a natural antimicrobial agent for food products.

  6. Effects of alkaline pretreatments and acid extraction conditions on the acid-soluble collagen from grass carp (Ctenopharyngodon idella) skin.

    PubMed

    Liu, Dasong; Wei, Guanmian; Li, Tiancheng; Hu, Jinhua; Lu, Naiyan; Regenstein, Joe M; Zhou, Peng

    2015-04-01

    This study investigated the effects of alkaline pretreatments and acid extraction conditions on the production of acid-soluble collagen (ASC) from grass carp skin. For alkaline pretreatment, 0.05 and 0.1M NaOH removed non-collagenous proteins without significant loss of ASC at 4, 10, 15 and 20 °C; while 0.2 and 0.5M NaOH caused significant loss of ASC, and 0.5M NaOH caused structural modification of ASC at 15 and 20 °C. For acid extraction at 4, 10, 15 and 20 °C, ASC was partly extracted by 0.1 and 0.2M acetic acid, while 0.5 and 1.0M acetic acid resulted in almost complete extraction. The processing conditions involving 0.05-0.1M NaOH for pretreatment, 0.5M acetic acid for extraction and 4-20 °C for both pretreatment and extraction, produced ASC with the structural integrity being well maintained and hence were recommended to prepare ASC from grass carp skin in practical application.

  7. A Standardized Protocol for Stereotaxic Intrahippocampal Administration of Kainic Acid Combined with Electroencephalographic Seizure Monitoring in Mice

    PubMed Central

    Bielefeld, Pascal; Sierra, Amanda; Encinas, Juan M.; Maletic-Savatic, Mirjana; Anderson, Anne; Fitzsimons, Carlos P.

    2017-01-01

    Lack of scientific reproducibility is a growing concern and weak experimental practices may contribute to irreproducibility. Here, we describe an optimized and versatile protocol for stereotaxic intrahippocampal administration of Kainic Acid (KA) in mice with a C57Bl6 background. In this protocol, KA administration is combined with in vivo recording of neuronal activity with wired and wireless setups. Following our protocol, KA administration results in a robust dose-dependent induction of low-level epileptiform activity or Status Epilepticus (SE) and induces previously characterized hallmarks of seizure-associated pathology. The procedure consists of three main steps: Craniotomy, stereotaxic administration of KA, and placement of recording electrodes in intrahippocampal, and subdural locations. This protocol offers extended possibilities compared to the systemic administration of KA, as it allows the researcher to accurately regulate the local dose of KA and resulting seizure activity, and permits the use and study of convulsive and non-convulsive KA doses, resulting in higher reproducibility and lower inter-individual variability and mortality rates. Caution should be taken when translating this procedure to different strains of mice as inter-strain sensitivity to KA has been described before. The procedure can be performed in ~1 h by a trained researcher, while intrahippocampal administration of KA without placing recording electrodes can be done in 25 min, and can be easily adapted to the titrated intrahippocampal administration of other drugs.

  8. Effects of a peracetic acid disinfection protocol on the biocompatibility and biomechanical properties of human patellar tendon allografts.

    PubMed

    Lomas, R J; Jennings, L M; Fisher, J; Kearney, J N

    2004-01-01

    Patellar tendon allografts, retrieved from cadaveric human donors, are widely used for replacement of damaged cruciate ligaments. In common with other tissue allografts originating from cadaveric donors, there are concerns regarding the potential for disease transmission from the donor to the recipient. Additionally, retrieval and subsequent processing protocols expose the graft to the risk of environmental contamination. For these reasons, disinfection or sterilisation protocols are necessary for these grafts before they are used clinically. A high-level disinfection protocol, utilising peracetic acid (PAA), has been developed and investigated for its effects on the biocompatibility and biomechanics of the patellar tendon allografts. PAA disinfection did not render the grafts either cytotoxic or liable to provoke an inflammatory response as assessed in vitro . However, the protocol was shown to increase the size of gaps between the tendon fibres in the matrix and render the grafts more susceptible to digestion with collagenase. Biomechanical studies of the tendons showed that PAA treatment had no effect on the ultimate tensile stress or Young's modulus of the tendons, and that ultimate strain was significantly higher in PAA treated tendons.

  9. Influence of hydrofluoric acid on extraction of thorium using a commercially available extraction chromatographic resin.

    PubMed

    Shimada-Fujiwara, Asako; Hoshi, Akiko; Kameo, Yutaka; Nakashima, Mikio

    2009-05-01

    The dependence of Th recovery on hydrofluoric acid (HF) concentration in nitric acid (HNO(3)) solutions (1-5 mol/dm(3)) containing 1x10(-6) mol/dm(3) of Th and various concentrations of HF and the elution behavior were studied using a commercially available UTEVA (for uranium and tetravalent actinide) resin column. Thorium recovery decreased with an increase in HF concentration in the sample solutions. The concentration of HF at which Th recovery started to decrease was approximately 1x10(-4) mol/dm(3) in 1 mol/dm(3) HNO(3) solution, approximately 1x10(-3) mol/dm(3) in 3 mol/dm(3) HNO(3) solution, and approximately 1x10(-2) mol/dm(3) in 5 mol/dm(3) HNO(3) solution. When Al(NO(3))(3) (0.2 mol/dm(3)) or Fe(NO(3))(3) (0.6 mol/dm(3)) was added as a masking agent for F(-) to the Th solution containing 1x10(-1) mol/dm(3) HF and 1 mol/dm(3) HNO(3), Th recovery improved from 1.4+/-0.3% to 95+/-5% or 93+/-3%. Effective extraction of Th using UTEVA resin was achieved by selecting the concentration of HNO(3) and/or adding masking agents such as Al(NO(3))(3) according to the concentration of HF in the sample solution.

  10. Ultrasound versus microwave as green processes for extraction of rosmarinic, carnosic and ursolic acids from rosemary.

    PubMed

    Jacotet-Navarro, M; Rombaut, N; Fabiano-Tixier, A-S; Danguien, M; Bily, A; Chemat, F

    2015-11-01

    Ultrasound and microwave as green processes are investigated in this study, focusing on the extraction selectivity towards antioxidant extraction from rosemary leaves. Due to its richness in valuable compounds such as rosmarinic, carnosic and ursolic acids, rosemary is a reference matrix for extraction study. In this work, six alternative processes are compared: ultrasound (bath, reactor and probe), microwave (reflux under microwave, microwave under nitrogen pressure and microwave under vapor pressure). The main result of this study is that selective extraction can be achieved according to extraction techniques and therefore to the extraction process.

  11. Extraction of ethanol with higher carboxylic acid solvents and their toxicity to yeast

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a screening exercise for ethanol-selective extraction solvents, partitioning of ethanol and water from a 5 wt% aqueous solution into several C8 – C18 carboxylic acids was studied. Results for the acids are compared with those from alcohols of similar structure. In all cases studied, the acids exh...

  12. Formic acid aided hot water extraction of hemicellulose from European silver birch (Betula pendula) sawdust.

    PubMed

    Goldmann, Werner Marcelo; Ahola, Juha; Mikola, Marja; Tanskanen, Juha

    2017-02-11

    Hemicellulose has been extracted from birch (Betula pendula) sawdust by formic acid aided hot water extraction. The maximum amount of hemicellulose extracted was about 70mol% of the total hemicellulose content at 170°C, measured as the combined yield of xylose and furfural. Lower temperatures (130 and 140°C) favored hemicellulose hydrolysis rather than cellulose hydrolysis, even though the total hemicellulose yield was less than at 170°C. It was found that formic acid greatly increased the hydrolysis of hemicellulose to xylose and furfural at the experimental temperatures. The amount of lignin in the extract remained below the detection limit of the analysis (3g/L) in all cases. Formic acid aided hot water extraction is a promising technique for extracting hemicellulose from woody biomass, while leaving a solid residue with low hemicellulose content, which can be delignified to culminate in the three main isolated lignocellulosic fractions: cellulose, hemicellulose, and lignin.

  13. Fatty acids and sterols composition, and antioxidant activity of oils extracted from plant seeds.

    PubMed

    Kozłowska, Mariola; Gruczyńska, Eliza; Ścibisz, Iwona; Rudzińska, Magdalena

    2016-12-15

    This study determined and compared the contents of bioactive components in plant seed oils extracted with n-hexane (Soxhlet method) and chloroform/methanol (Folch method) from coriander, caraway, anise, nutmeg and white mustard seeds. Oleic acid dominated among unsaturated fatty acids in nutmeg and anise seed oils while petroselinic acid was present in coriander and caraway oils. Concerning sterols, β-sitosterol was the main component in seed oils extracted with both methods. The content of total phenolics in nutmeg, white mustard and coriander seed oils extracted with chloroform/methanol was higher than in their counterparts prepared with n-hexane. The seed oil samples extracted according to the Folch method exhibited a higher ability to scavenge DPPH radicals compared to the oil samples prepared with the Soxhlet method. DPPH values of the methanolic extracts derived from oils produced with the Folch method were also higher than in the oils extracted with n-hexane.

  14. Removal of acetic acid from simulated hemicellulosic hydrolysates by emulsion liquid membrane with organophosphorus extractants.

    PubMed

    Lee, Sang Cheol

    2015-09-01

    Selective removal of acetic acid from simulated hemicellulosic hydrolysates containing xylose and sulfuric acid was attempted in a batch emulsion liquid membrane (ELM) system with organophosphorus extractants. Various experimental variables were used to develop a more energy-efficient ELM process. Total operation time of an ELM run with a very small quantity of trioctylphosphine oxide as the extractant was reduced to about a third of those required to attain almost the same extraction efficiency as obtained in previous ELM works without any extractant. Under specific conditions, acetic acid was selectively separated with a high degree of extraction and insignificant loss of xylose, and its purity and enrichment ratio in the stripping phase were higher than 92% and 6, respectively. Also, reused organic membrane solutions exhibited the extraction efficiency as high as fresh organic solutions did. These results showed that the current ELM process would be quite practical.

  15. Copper extraction by fatty hydroxamic acids derivatives synthesized based on palm kernel oil.

    PubMed

    Haron, Jelas; Jahangirian, Hossein; Silong, Sidik; Yusof, Nor Azah; Kassim, Anuar; Moghaddam, Roshanak Rafiee; Peyda, Mazyar; Abdollahi, Yadollah; Amin, Jamileh; Gharayebi, Yadollah

    2012-01-01

    Fatty hydroxamic acids derivatives based on palm kernel oil which are phenyl fatty hydroxamic acids (PFHAs), methyl fatty hydroxamic acids (MFHAs), isopropyl fatty hydroxamic acids (IPFHAs) and benzyl fatty hydroxamic acids (BFHAs) were applied as chelating agent for copper liquid-liquid extraction. The extraction of copper from aqueous solution by MFHAs, PFHAs, BFHAs or IPFHAs were carried out in hexane as an organic phase through the formation of copper methyl fatty hydroxamate (Cu-MFHs), copper phenyl fatty hydroxamate (Cu-PFHs), copper benzyl fatty hydroxamate (Cu-BFHs) and copper isopropyl fatty hydroxamate (Cu-IPFHs). The results showed that the fatty hydroxamic acid derivatives could extract copper at pH 6.2 effectively with high percentage of extraction (the percentages of copper extraction by MFHAs, PFHAs, IPFHs and BFHAs were found to be 99.3, 87.5, 82.3 and 90.2%, respectively). The extracted copper could be quantitatively stripped back into sulphuric acid (3M) aqueous solution. The obtained results showed that the copper recovery percentages from Cu-MFHs, Cu-PFHs, Cu-BFHs and Cu-IPFHs are 99.1, 99.4, 99.6 and 99.9 respectively. The copper extraction was not affected by the presence of a large amount of Mg (II), Ni (II), Al (III), Mn (II) and Co (II) ions in the aqueous solution.

  16. Solid-phase extraction of betanin and isobetanin from beetroot extracts using a dipicolinic acid molecularly imprinted polymer.

    PubMed

    Nestora, Sofia; Merlier, Franck; Prost, Elise; Haupt, Karsten; Rossi, Claire; Tse Sum Bui, Bernadette

    2016-09-23

    Betanin is a natural pigment with significant antioxidant and biological activities currently used as food colorant. The isolation of betanin is problematic due to its instability. In this work, we developed a fast and economic procedure based on molecularly imprinted solid-phase extraction (MISPE) for the selective clean-up of betanin and its stereoisomer isobetanin from beetroot extracts. Dipicolinic acid was used as template for the MIP preparation because of its structural similarity with the chromophore group of betanin. The MISPE procedures were fully optimized allowing the almost complete removal of matrix components such as sugars and proteins, resulting in high extraction recovery of betanin/isobetanin in a single step. Moreover, the whole extraction procedure was performed in environmentally friendly solvents with either ethanol or water. Our MISPE method is very promising for the future development of well-formulated beetroot extract with specified betanin/isobetanin content, ready for food or medicinal use.

  17. Molecular tools and protocols for engineering the acid-tolerant yeast Zygosaccharomyces bailii as a potential cell factory.

    PubMed

    Branduardi, Paola; Dato, Laura; Porro, Danilo

    2014-01-01

    Microorganisms offer a tremendous potential as cell factories, and they are indeed used by humans for centuries for biotransformations. Among them, yeasts combine the advantage of unicellular state with a eukaryotic organization, and, in the era of biorefineries, their biodiversity can offer solutions to specific process constraints. Zygosaccharomyces bailii, an ascomycetales budding yeast, is widely known for its peculiar tolerance to various stresses, among which are organic acids. Despite the possibility to apply with this yeast some of the molecular tools and protocols routinely used to manipulate Saccharomyces cerevisiae, adjustments and optimizations are necessary. Here, we describe in detail protocols for transformation, for target gene disruption or gene integration, and for designing episomal expression plasmids helpful for developing and further studying the yeast Z. bailii.

  18. Simultaneous determination of docosahexaenoic acid and eicosapentaenoic acid in common seafood using ultrasonic cell crusher extraction combined with gas chromatography.

    PubMed

    Zhao, Juanjuan; Ren, Yan; Yu, Chen; Chen, Xiangming; Shi, Yanan

    2017-02-01

    An effective method for the simultaneous determination of docosahexaenoic acid and eicosapentaenoic acid in common seafood by gas chromatography was developed and validated. Total docosahexaenoic acid and eicosapentaenoic acid were extracted from seafood by ultrasonic cell crusher assisted extraction and methyl esterified for gas chromatography analysis in the presence of the internal standard. The linearity was good (r > 0.999) in 9.59 ∼ 479.5 μg/mL for docosahexaenoic acid and 9.56 ∼ 477.8 μg/mL for eicosapentaenoic acid. The intrarun and interrun precisions were both within 4.8 and 6.1% for the two analytes, while the accuracy was less than 5.8%. The developed method was applied for determination of docosahexaenoic acid and eicosapentaenoic acid in six kinds of seafood. The result showed the content of docosahexaenoic acid and eicosapentaenoic acid was all higher than 1 mg/g in yellow croaker, hairtail, venerupis philippinarum, mussel, and oyster. Our work may be helpful for dietary optimization and production of docosahexaenoic acid and eicosapentaenoic acid.

  19. Synchronous determination with double-wavelength by RP-HPLC-UV and optimization of ultrasound-assisted extraction of phenolic acids from Caragana species using response surface methodology.

    PubMed

    Zeng, Zhi; Ji, Zhongyin; Hu, Na; Chen, Shasha; Bai, Bo; Wang, Honglun; Suo, Yourui

    2017-03-14

    The utilization of Caragana korshinskii Kom. (CK) is currently concentrated on its ecological and fuel functions. Little attention has been devoted to the analysis of their phenolic acid (PA) components. To obtain more data for further utilization of CK, a new analysis protocol was tested to determine PAs synchronously by RP-HPLC-UV with double-wavelength (280nm and 320nm) detection. Specifically, separation of PA components was performed on a Hypersil Gold C18 reverse phase column with gradient elution. A four-factor-three-level Box-Behnken design was implemented for optimization of PA extraction. The results demonstrated that CK were rich primarily in chlorogenic acid, vanillic acid, caffeic acid and rosmarinic acid. The total content of PAs in CK leaves was the highest compared with its other parts. The distribution of total flavonoid content of CK was leaves>flowers>bark, while that of the total phenolic content of CK was flowers>leaves>bark.

  20. An Evaluation of Partial Digestion Protocols for the Extraction and Measurement of Trace Metals of Environmental Concern in Marine and Estuarine Sediments

    NASA Astrophysics Data System (ADS)

    Winters, S. J.; Krahforst, C.; Sherman, L.; Kehm, K.

    2013-12-01

    As part of a broad study of the fate and transport of trace metals in estuarine sediments (Krahforst et al., 2013), the efficacy of commonly-used partial digestion protocols, including ISO 11466 (treatment with aqua regia), EPA 3050B (nitric acid followed by H2O2) and a modified rock digestion method ('RD' method- H2O2 followed by nitric), were evaluated for two NIST SRM materials, marine sediment 2702 and estuarine sediment 1646a. Unlike so-called total sediment digestions, the methods studied in this work do not employ hydrofluoric acid and are thought to leave silicates substantially or wholly intact. These methods can in principle compliment studies based on total digestions by providing information about trace metals in phases that are potentially more labile in the marine environment. Samples were digested in ~150 mg aliquots. Application of ISO 11466 and EPA 3050B followed published protocols except that digestions were carried out in trace-metal clean 15 mL capped Teflon vessels in an Al block digester and, at the end of the procedure, the supernatant was decanted from undigested material following repeated centrifugation in 2% nitric acid. Digested solutions were analyzed for Al, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Ag, Cd, Sn and Pb content by ICPMS. All elements were analyzed in collision reaction cell mode to minimize isobaric interferences, except Cd and Ag, which were analyzed in standard mode. Instrument performance was monitored in-run by analyzing the SRM 1643e and several quality-check standards. Two repeated digestions of SRM 2702 and SRM 1646a using EPA 3050B produced identical yields, within the standard deviation of repeated analyses (0 - 5%), for all analyzed elements except Cu, which varied by 30% for SRM 2702. The same was true for ISO 11466, although the standard deviation of repeated analyses for this digestion series tended to be larger (< ~15%). The RD method, which consists of pre-treatment with H2O2 followed by repeated treatments with

  1. Role of deoxyribonucleic acid ligase in a doxyribonucleic acid membrane fraction extracted from pneumococci.

    PubMed Central

    Greene, M; Firshein, W

    1976-01-01

    Deoxyribonucleic acid (DNA) ligase has been detected in a DNA membrane fraction extracted from Pneumococcus. The specific activity of the enzyme in this fraction is 10-fold greater than in the remaining cell extract. It remains firmly bound (with other enzymes) to the complex after a purification procedure in which a considerable percentage of the macromolecules are dissociated. The ligase acts in two ways in the DNA membrane fraction in vitro. One, it catalyzes the linkage of small-molecular-weight pieces of newly synthesized DNA into heavier-molecular-weight DNA strands as shown by others (M Gellert, 1976; R. Okazaki, A. Sugino, S. Hirose, T. Okazaki, Y. Imae, R. Kainuma-Kuroda, T. Ogawa, M. Arisawa, and Y. Kurosowa, 1973; B. Olivera and I. Lehman, 14; and A. Sugino, S. Hirose, and R. Okazaki, 1972) and, two, it protects DNA from degradation by deoxyribonucleases. This latter effect is due to a competition between the ability of the nucleases to degrade DNA and the ability of DNA ligase to seal the nicks produced by these degradative enzymes. The ligase acts cooperatively with other enzymes in the DNA membrane fraction to synthesize DNA. PMID:4433

  2. Analysis of several methods for the extraction of high quality DNA from acetic acid bacteria in wine and vinegar for characterization by PCR-based methods.

    PubMed

    Jara, C; Mateo, E; Guillamón, J M; Torija, M J; Mas, A

    2008-12-10

    Acetic acid bacteria (AAB) are fastidious microorganisms with poor recovery in culture. Culture-independent methods are currently under examination. Good DNA extraction is a strict requirement of these methods. We compared five methods for extracting the DNA of AAB directly from wine and vinegar samples. Four matrices (white wine, red wine, superficial vinegar and submerged vinegar) contaminated with two AAB strains belonging to Acetobacter pasteurianus and Gluconacetobacter hansenii were assayed. To improve the yield and quality of the extracted DNA, a sample treatment (washing with polyvinyl pyrrolidone or NaCl) was also tested. DNA quality was measured by amplification of the 16S rRNA gene with conventional PCR. DNA recovery rate was assessed by real-time PCR. DNA amplification was always successful with the Wizard method though DNA recovery was poor. A CTAB-based method and NucleoSpin protocol extracted the highest DNA recoveries from wine and vinegar samples. Both of these methods require treatment to recover suitable DNA for amplification with maximum recovery. Both may therefore be good solutions for DNA extraction in wine and vinegar samples. DNA extraction of Ga hansenii was more effective than that of A. pasteurianus. The fastest and cheapest method we evaluated (the Thermal shock protocol) produced the worst results both for DNA amplification and DNA recovery.

  3. Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures.

    PubMed

    Minas, Konstantinos; McEwan, Neil R; Newbold, Charles Jamie; Scott, Karen P

    2011-12-01

    The quality and yield of extracted DNA are critical for the majority of downstream applications in molecular biology. Moreover, molecular techniques such as quantitative real-time PCR (qPCR) are becoming increasingly widespread; thus, validation and cross-laboratory comparison of data require standardization of upstream experimental procedures. DNA extraction methods depend on the type and size of starting material(s) used. As such, the extraction of template DNA is arguably the most significant variable when cross-comparing data from different laboratories. Here, we describe a reliable, inexpensive and rapid method of DNA purification that is equally applicable to small or large scale or high-throughput purification of DNA. The protocol relies on a CTAB-based buffer for cell lysis and further purification of DNA with phenol : chloroform : isoamyl alcohol. The protocol has been used successfully for DNA purification from rumen fluid and plant cells. Moreover, after slight alterations, the same protocol was used for large-scale extraction of DNA from pure cultures of Gram-positive and Gram-negative bacteria. The yield of the DNA obtained with this method exceeded that from the same samples using commercial kits, and the quality was confirmed by successful qPCR applications.

  4. Tachikawa project for prevention of posttraumatic stress disorder with polyunsaturated fatty acid (TPOP): study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background Preclinical and clinical studies suggest that supplementation with omega-3 fatty acids after trauma might reduce subsequent posttraumatic stress disorder (PTSD). To date, we have shown in an open trial that PTSD symptoms in critically injured patients can be reduced by taking omega-3 fatty acids, hypothesized to stimulate hippocampal neurogenesis. The primary aim of the present randomized controlled trial is to examine the efficacy of omega-3 fatty acid supplementation in the secondary prevention of PTSD following accidental injury, as compared with placebo. This paper describes the rationale and protocol of this trial. Methods/design The Tachikawa Project for Prevention of Posttraumatic Stress Disorder with Polyunsaturated Fatty Acid (TPOP) is a double-blinded, parallel group, randomized controlled trial to assess whether omega-3 fatty acid supplementation can prevent PTSD symptoms among accident-injured patients consecutively admitted to an intensive care unit. We plan to recruit accident-injured patients and follow them prospectively for 12 weeks. Enrolled patients will be randomized to either the omega-3 fatty acid supplement group (1,470 mg docosahexaenoic acid and 147 mg eicosapentaenoic acid daily) or placebo group. Primary outcome is score on the Clinician-Administered PTSD Scale (CAPS). We will need to randomize 140 injured patients to have 90% power to detect a 10-point difference in mean CAPS scores with omega-3 fatty acid supplementation compared with placebo. Secondary measures are diagnosis of PTSD and major depressive disorder, depressive symptoms, physiologic response in the experiment using script-driven imagery and acoustic stimulation, serum brain-derived neurotrophic factor, health-related quality of life, resilience, and aggression. Analyses will be by intent to treat. The trial was initiated on December 13 2008, with 104 subjects randomized by November 30 2012. Discussion This study promises to be the first trial to provide a novel

  5. Determining the fatty acid composition in plasma and tissues as fatty acid methyl esters using gas chromatography – a comparison of different derivatization and extraction procedures.

    PubMed

    Ostermann, Annika I; Müller, Maike; Willenberg, Ina; Schebb, Nils Helge

    2014-12-01

    Analysis of the fatty acid (FA) composition in biological samples is commonly carried out using gas liquid chromatography (GC) after transesterification to volatile FA methyl esters (FAME). We compared the efficacy of six frequently used protocols for derivatization of different lipid classes as well as for plasma and tissue samples. Transesterification with trimethylsulfonium hydroxide (TMSH) led to insufficient derivatization efficacies for polyunsaturated FAs (PUFA, <50%). Derivatization in presence of potassium hydroxide (KOH) failed at derivatizing free FAs (FFAs). Boron trifluoride (BF3) 7% in hexane/MeOH (1:1) was insufficient for the transesterification of cholesterol ester (CE) as well as triacylglycerols (TGs). In contrast, methanolic hydrochloric acid (HCl) as well as a combination of BF3 with methanolic sodium hydroxide (NaOH+BF3) were suitable for the derivatization of FFAs, polar lipids, TGs, and CEs (derivatization rate >80% for all tested lipids). Regarding plasma samples, all methods led to an overall similar relative FA pattern. However, significant differences were observed, for example, for the relative amount of EPA+DHA (n3-index). Absolute FA plasma concentrations differed considerably among the methods, with low yields for KOH and BF3. We also demonstrate that lipid extraction with tert-butyl methyl ether/methanol (MTBE/MeOH) is as efficient as the classical method according to Bligh and Dyer, making it possible to replace (environmentally) toxic chloroform.We conclude that HCl-catalyzed derivatization in combination with MeOH/MTBE extraction is the most appropriate among the methods tested for the analysis of FA concentrations and FA pattern in small biological samples. A detailed protocol for the analysis of plasma and tissues is included in this article.

  6. COMBINING NEUTRAL AND ACIDIC EXTRACTANTS FOR RECOVERING TRANSURANIC ELEMENTS FROM NUCLEAR FUEL

    SciTech Connect

    Lumetta, Gregg J.; Neiner, Doinita; Sinkov, Sergey I.; Carter, Jennifer C.; Braley, Jenifer C.; Latesky, Stanley; Gelis, Artem V.; Tkac, Peter; Vandegrift, George F.

    2011-10-03

    We have been investigating a solvent extraction system that combines a neutral extractant--octyl(phenyl)-N,N-diisobutyl-carbamoylmethylphosphine oxide (CMPO)--with an acidic extractant--bis(2-ethylhexyl)phosphoric acid (HDEHP)--to form a single process solvent for separating Am and Cm from the other components of irradiated nuclear fuel. It was originally hypothesized that the extraction chemistry of CMPO would dominate under conditions of high acidity (> 1 M HNO3), resulting in co-extraction of the transuranic and lanthanide elements into the organic phase. Contacting the loaded solvent with a solution of diethylenetriaminepentaacetate (DTPA) buffered with lactic or citric acid at pH {approx}3 to 4 would result in a condition in which the HDEHP chemistry dominates. Although the data somewhat support this hypothesis, it is clear that there are interactions between the two extractants such that they do not act independently in the extraction and stripping regimes. We report here studies directed at determining the nature and extent of interaction between CMPO and HDEHP, the synergistic behavior of CMPO and HDEHP in the extraction of americium and neodymium, and progress towards determining the thermodynamics of this extraction system. Neodymium and americium behave similarly in the combined solvent system, with a significant synergy between CMPO and HDEHP in the extraction of both of these trivalent elements from lactate-buffered DTPA solutions. In contrast, a much weaker synergistic behaviour is observed for europium. Thus, investigations into the fundamental chemistry involved in this system have focused on the neodymium extraction. The extraction of neodymium has been systematically investigated, individually varying the HDEHP concentration, the CMPO concentration, or the aqueous phase composition. Thermodynamic modeling of the neodymium extraction system has been initiated. Interactions between CMPO and HDEHP in the organic phase must be taken into account in

  7. Chemical composition of phase I Coxiella burnetii soluble antigen prepared by trichloroacetic acid extraction.

    PubMed

    Lukácová, M; Brezina, R; Schramek, S; Pastorek, J

    1989-01-01

    Optimal conditions of extraction (time and temperature) by trichloroacetic acid of soluble antigen from phase I Coxiella burnetii (TCAE), possessing protective properties and used as a chemovaccine against Q fever in men, were studied. Extracts prepared under various conditions were analysed for their polysaccharide, protein and phosphorus contents. Forty-five min of extraction at 0 degrees C were sufficient to obtain a soluble antigen reacting in immunodiffusion with hyperimmune rabbit antiserum. The polysaccharide contents decreased with prolonged extraction at 0 degrees C. At higher extraction temperatures (37 and 100 degrees C), the polysaccharide contents increased while that of proteins decreased. TCAE prepared at 100 degrees C gave no positive immunodiffusion reaction.

  8. Extraction of gallium(III) from hydrochloric acid solutions by trioctylammonium-based mixed ionic liquids.

    PubMed

    Katsuta, Shoichi; Okai, Miho; Yoshimoto, Yuki; Kudo, Yoshihiro

    2012-01-01

    The extractabilities of aluminium(III), gallium(III), and indium(III) from hydrochloric acid solutions were investigated using a mixture of two protic ionic liquids, trioctylammonium bis(trifluoromethanesulfonyl)amide ([TOAH][NTf(2)]) and trioctylammonium nitrate ([TOAH][NO(3)]). At a HCl concentration of 4 mol L(-1) or more, gallium(III) was nearly quantitatively extracted and the extractability order was Ga > Al > In. The extractability of gallium(III) increased with increasing [TOAH][NO(3)] content in the mixed ionic liquid. The extracted gallium(III) was quantitatively stripped with aqueous nitric acid solutions. The separation and recovery of gallium(III) from hydrochloric acid solutions containing excess indium(III) was demonstrated using the mixed ionic liquid.

  9. Simultaneous determination of gibberellic acid, indole-3-acetic acid and abscisic acid in wheat extracts by solid-phase extraction and liquid chromatography-electrospray tandem mass spectrometry.

    PubMed

    Hou, Shengjie; Zhu, Jiang; Ding, Mingyu; Lv, Guohua

    2008-08-15

    A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for simultaneous determination of three representative phytohormones in plant samples: gibberellic acid (GA(3)), indole-3-acetic acid (IAA) and abscisic acid (ABA). A solid-phase extraction (SPE) pretreatment method was used to concentrate and purify the three phytohormones of different groups from plant samples. The separation was carried out on a C(18) reversed-phase column, using methanol/water containing 0.2% formic acid (50:50, v/v) as the isocratic mobile phase at the flow-rate of 1.0 mL min(-1), and the three phytohormones were eluted within 7 min. A linear ion trap mass spectrometer equipped with electrospray ionization source was operated in negative ion mode. Selective reaction monitoring (SRM) was employed for quantitative measurement. The SRM transitions monitored were as 345-->239, 301 for GA(3), 174-->130 for IAA and 263-->153, 219 for ABA. Good linearities were found within the ranges of 5-200 microg mL(-1) for IAA and 0.005-10 microg mL(-1) for ABA and GA(3). Their detection limits based on a signal-to-noise ratio of three were 0.005 microg mL(-1), 2.2 microg mL(-1) and 0.003 microg mL(-1) for GA(3), IAA and ABA, respectively. Good recoveries from 95.5% to 102.4% for the three phytohormones were obtained. The results demonstrated that the SPE-LC-MS/MS method developed is highly effective for analyzing trace amounts of the three phytohormones in plant samples.

  10. A solvent extraction approach to recover acetic acid from mixed waste acids produced during semiconductor wafer process.

    PubMed

    Shin, Chang-Hoon; Kim, Ju-Yup; Kim, Jun-Young; Kim, Hyun-Sang; Lee, Hyang-Sook; Mohapatra, Debasish; Ahn, Jae-Woo; Ahn, Jong-Gwan; Bae, Wookeun

    2009-03-15

    Recovery of acetic acid (HAc) from the waste etching solution discharged from silicon wafer manufacturing process has been attempted by using solvent extraction process. For this purpose 2-ethylhexyl alcohol (EHA) was used as organic solvent. In the pre-treatment stage >99% silicon and hydrofluoric acid was removed from the solution by precipitation. The synthesized product, Na(2)SiF(6) having 98.2% purity was considered of commercial grade having good market value. The waste solution containing 279 g/L acetic acid, 513 g/L nitric acid, 0.9 g/L hydrofluoric acid and 0.030 g/L silicon was used for solvent extraction study. From the batch test results equilibrium conditions for HAc recovery were optimized and found to be 4 stages of extraction at an organic:aqueous (O:A) ratio of 3, 4 stages of scrubbing and 4 stages of stripping at an O:A ratio of 1. Deionized water (DW) was used as stripping agent to elute HAc from organic phase. In the whole batch process 96.3% acetic acid recovery was achieved. Continuous operations were successfully conducted for 100 h using a mixer-settler to examine the feasibility of the extraction system for its possible commercial application. Finally, a complete process flowsheet with material balance for the separation and recovery of HAc has been proposed.

  11. Extracting metal ions with diphosphonic acid, or derivative thereof

    DOEpatents

    Horwitz, Earl P.; Gatrone, Ralph C.; Nash, Kenneth L.

    1994-01-01

    Thermodynamically-unstable complexing agents which are diphosphonic acids and diphosphonic acid derivatives (or sulphur containing analogs), like carboxyhydroxymethanediphosphonic acid and vinylidene-1,1-diphosphonic acid, are capable of complexing with metal ions, and especially metal ions in the II, III, IV, V and VI oxidation states, to form stable, water-soluble metal ion complexes in moderately alkaline to highly-acidic media. However, the complexing agents can be decomposed, under mild conditions, into non-organic compounds which, for many purposes are environmentally-nondamaging compounds thereby degrading the complex and releasing the metal ion for disposal or recovery. Uses for such complexing agents as well as methods for their manufacture are also described.

  12. Membrane extraction with thermodynamically unstable diphosphonic acid derivatives

    DOEpatents

    Horwitz, Earl Philip; Gatrone, Ralph Carl; Nash, Kenneth LaVerne

    1997-01-01

    Thermodynamically-unstable complexing agents which are diphosphonic acids and diphosphonic acid derivatives (or sulphur containing analogs), like carboxyhydroxymethanediphosphonic acid and vinylidene-1,1-diphosphonic acid, are capable of complexing with metal ions, and especially metal ions in the II, III, IV, V and VI oxidation states, to form stable, water-soluble metal ion complexes in moderately alkaline to highly-acidic media. However, the complexing agents can be decomposed, under mild conditions, into non-organic compounds which, for many purposes are environmentally-nondamaging compounds thereby degrading the complex and releasing the metal ion for disposal or recovery. Uses for such complexing agents as well as methods for their manufacture are also described.

  13. Membrane extraction with thermodynamically unstable diphosphonic acid derivatives

    DOEpatents

    Horwitz, E.P.; Gatrone, R.C.; Nash, K.L.

    1997-10-14

    Thermodynamically-unstable complexing agents which are diphosphonic acids and diphosphonic acid derivatives (or sulphur containing analogs), like carboxyhydroxymethanediphosphonic acid and vinylidene-1,1-diphosphonic acid, are capable of complexing with metal ions, and especially metal ions in the II, III, IV, V and VI oxidation states, to form stable, water-soluble metal ion complexes in moderately alkaline to highly-acidic media. However, the complexing agents can be decomposed, under mild conditions, into non-organic compounds which, for many purposes are environmentally-nondamaging compounds thereby degrading the complex and releasing the metal ion for disposal or recovery. Uses for such complexing agents as well as methods for their manufacture are also described. 1 fig.

  14. Extracting metal ions with diphosphonic acid, or derivative thereof

    DOEpatents

    Horwitz, E.P.; Gatrone, R.C.; Nash, K.L.

    1994-07-26

    Thermodynamically-unstable complexing agents which are diphosphonic acids and diphosphonic acid derivatives (or sulfur containing analogs), like carboxyhydroxymethanediphosphonic acid and vinylidene-1,1-diphosphonic acid, are capable of complexing with metal ions, and especially metal ions in the II, III, IV, V and VI oxidation states, to form stable, water-soluble metal ion complexes in moderately alkaline to highly-acidic media. However, the complexing agents can be decomposed, under mild conditions, into non-organic compounds which, for many purposes are environmentally-nondamaging compounds thereby degrading the complex and releasing the metal ion for disposal or recovery. Uses for such complexing agents as well as methods for their manufacture are also described. 1 fig.

  15. Methods for extraction and determination of phenolic acids in medicinal plants: a review.

    PubMed

    Arceusz, Agnieszka; Wesolowski, Marek; Konieczynski, Pawel

    2013-12-01

    Phenolic acids constitute a group of potentially immunostimulating compounds. They occur in all medicinal plants and are widely used in phytotherapy and foods of plant origin. In recent years, phenolic acids have attracted much interest owing to their biological functions. This paper reviews the extraction and determination methods of phenolic acids in medicinal plants over the last 10 years. Although Soxhlet extraction and ultrasonic assisted extraction (UAE) are commonly used for the extraction of phenolic acids from plant materials, alternative techniques such as supercritical fluid extraction (SFE), and accelerated solvent extraction (ASE) can also be used. After extraction, phenolic acids are determined usually by liquid chromatography (LC) owing to the recent developments in this technique, especially when it is coupled with mass spectrometry (MS). Also detection systems are discussed, including UV-Vis, diode array, electrochemical and fluorimetric. Other popular techniques for the analysis of this group of secondary metabolites are gas chromatography coupled with mass spectrometry (GC-MS) and capillary electrophoresis (CE).

  16. Some Antifungal Properties of Sorbic Acid Extracted from Berries of Rowan (Sorbus Aucuparia).

    ERIC Educational Resources Information Center

    Brunner, Ulrich

    1985-01-01

    The food preservative sorbic acid can be extracted from Eurasian mountain ash berries (commercially available) and used to show antifungal properties in microbiological investigations. Techniques for extraction, purification, ultraviolet analysis, and experiments displaying antifungal activity are described. A systematic search for similar…

  17. 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…

  18. Nanofiltration, bipolar electrodialysis and reactive extraction hybrid system for separation of fumaric acid from fermentation broth.

    PubMed

    Prochaska, Krystyna; Staszak, Katarzyna; Woźniak-Budych, Marta Joanna; Regel-Rosocka, Magdalena; Adamczak, Michalina; Wiśniewski, Maciej; Staniewski, Jacek

    2014-09-01

    A novel approach based on a hybrid system allowing nanofiltration, bipolar electrodialysis and reactive extraction, was proposed to remove fumaric acid from fermentation broth left after bioconversion of glycerol. The fumaric salts can be concentrated in the nanofiltration process to a high yield (80-95% depending on pressure), fumaric acid can be selectively separated from other fermentation components, as well as sodium fumarate can be conversed into the acid form in bipolar electrodialysis process (stack consists of bipolar and anion-exchange membranes). Reactive extraction with quaternary ammonium chloride (Aliquat 336) or alkylphosphine oxides (Cyanex 923) solutions (yield between 60% and 98%) was applied as the final step for fumaric acid recovery from aqueous streams after the membrane techniques. The hybrid system permitting nanofiltration, bipolar electrodialysis and reactive extraction was found effective for recovery of fumaric acid from the fermentation broth.

  19. Application of dissolvable layered double hydroxides as sorbent in dispersive solid-phase extraction and extraction by co-precipitation for the determination of aromatic acid anions.

    PubMed

    Tang, Sheng; Lee, Hian Kee

    2013-08-06

    Three types of magnesium-aluminum layered double hydroxides were synthesized and employed as solid-phase extraction (SPE) sorbents to extract several aromatic acids (protocatechuic acid, mandelic acid, phthalic acid, benzoic acid, and salicylic acid) from aqueous samples. An interesting feature of these sorbents is that they dissolve when the pH of the solution is lower than 4. Thus, the analyte elution step, as needed in conventional sorbent-based extraction, was obviated by dissolving the sorbent in acid after extraction and separation from the sample solution. The extract was then directly injected into a high-performance liquid chromatography-ultraviolet detection system for analysis. In the key adsorption process, both dispersive SPE and co-precipitation extraction with the sorbents were conducted and experimental parameters such as pH, temperature, and extraction time were optimized. The results showed that both extraction methods provided low limits of detection (0.03-1.47 μg/L) and good linearity (r(2) > 0.9903). The optimized extraction conditions were applied to human urine and sports drink samples. This new and interesting extraction approach was demonstrated to be a fast and efficient procedure for the extraction of organic anions from aqueous samples.

  20. Simple DNA extraction protocol for a 16S rDNA study of bacterial diversity in tropical landfarm soil used for bioremediation of oil waste.

    PubMed

    Maciel, B M; Santos, A C F; Dias, J C T; Vidal, R O; Dias, R J C; Gross, E; Cascardo, J C M; Rezende, R P

    2009-03-31

    Landfarm soil is used to bioremediate oil wastes from petrochemical industries. We developed a simplified protocol for microbial DNA extraction of tropical landfarm soil using only direct lysis of macerated material. Two samples of tropical landfarm soil from a Brazilian refinery were analyzed by this protocol (one consisted of crude oil-contaminated soil; the other was continuously enriched for nine months with petroleum). The soil samples were lysed by maceration with liquid nitrogen, eliminating the need for detergents, organic solvents and enzymatic cell lysis. Then, the DNA from the lysed soil sample was extracted using phenol-chloroform-isoamyl alcohol or guanidium isothiocyanate, giving high DNA yields (more than 1 micro g DNA/g soil) from both soil types. This protocol compared favorably with an established method of DNA template preparation that included mechanical, chemical and enzymatic treatment for cell lysis. The efficiency of this extraction protocol was confirmed by polymerase chain reaction amplification of the 16S rRNA gene, denaturing gradient gel electrophoresis and cloning assays. Fifty-one different clones were obtained; their sequences were classified into at least seven different phyla of the Eubacteria group (Proteobacteria - alpha, gamma and delta, Chloroflexi, Actinobacteria, Acidobac teria, Planctomycetes, Bacteroidetes, and Firmicutes). Forty percent of the sequences could not be classified into these phyla, demonstrating the genetic diversity of this microbial community. Only eight isolates had sequences similar to known sequences of 16S rRNA of cultivable organisms or of known environmental isolates and therefore could be identified to the genus level. This method of DNA extraction is a useful tool for analysis of the bacteria responsible for petroleum degradation in contaminated environments.

  1. An accessible protocol for solid-phase extraction of N-linked glycopeptides through reductive amination by amine-functionalized magnetic nanoparticles.

    PubMed

    Zhang, Ying; Kuang, Min; Zhang, Lijuan; Yang, Pengyuan; Lu, Haojie

    2013-06-04

    In light of the significance of glycosylation for wealthy biological events, it is important to prefractionate glycoproteins/glycopeptides from complex biological samples. Herein, we reported a novel protocol of solid-phase extraction of glycopeptides through a reductive amination reaction by employing the easily accessible 3-aminopropyltriethoxysilane (APTES)-functionalized magnetic nanoparticles. The amino groups from APTES, which were assembled onto the surface of the nanoparticles through a one-step silanization reaction, could conjugate with the aldehydes from oxidized glycopeptides and, therefore, completed the extraction. To the best of our knowledge, this is the first example of applying the reductive amination reaction into the isolation of glycopeptides. Due to the elimination of the desalting step, the detection limit of glycopeptides was improved by 2 orders of magnitude, compared to the traditional hydrazide chemistry-based solid phase extraction, while the extraction time was shortened to 4 h, suggesting the high sensitivity, specificity, and efficiency for the extraction of N-linked glycopeptides by this method. In the meantime, high selectivity toward glycoproteins was also observed in the separation of Ribonuclease B from the mixtures contaminated with bovine serum albumin. What's more, this technique required significantly less sample volume, as demonstrated in the successful mapping of glycosylation of human colorectal cancer serum with the sample volume as little as 5 μL. Because of all these attractive features, we believe that the innovative protocol proposed here will shed new light on the research of glycosylation profiling.

  2. A General Protocol for the Polycondensation of Thienyl N-Methyliminodiacetic Acid Boronate Esters To Form High Molecular Weight Copolymers

    PubMed Central

    2016-01-01

    Thienyl di-N-methyliminodiacetic acid (MIDA) boronate esters are readily synthesized by electrophilic C–H borylation producing bench stable crystalline solids in good yield and excellent purity. Optimal conditions for the slow release of the boronic acid using KOH as the base in biphasic THF/water mixtures enables the thienyl MIDA boronate esters to be extremely effective homo-bifunctionalized (AA-type) monomers in Suzuki–Miyaura copolymerizations with dibromo-heteroarenes (BB-type monomers). A single polymerization protocol is applicable for the formation of five alternating thienyl copolymers that are (or are close analogues of) state of the art materials used in organic electronics. The five polymers were produced in excellent yields and with high molecular weights comparable to those produced using Stille copolymerization protocols. Therefore, thienyl di-MIDA boronate esters represent bench stable and low toxicity alternatives to highly toxic di-trimethylstannyl AA-type monomers that are currently ubiquitous in the synthesis of these important alternating copolymers. PMID:27636745

  3. Hydroxycinnamic Acid Derivatives Obtained from a Commercial Crataegus Extract and from Authentic Crataegus spp.§

    PubMed Central

    Kuczkowiak, Ulrich; Petereit, Frank; Nahrstedt, Adolf

    2014-01-01

    Abstract Eleven hydroxycinnamic acid derivatives were isolated from a 70% methanolic Crataegus extract (Crataegi folium cum flore) and partly verified and quantified for individual Crataegus species (C. laevigata, C. monogyna, C. nigra, C. pentagyna) by HPLC: 3-O-(E)-p-coumaroylquinic acid (1), 5-O-(E)-p-coumaroyl-quinic acid (2), 4-O-(E)-p-coumaroylquinic acid (3), 3-O-(E)-caffeoylquinic acid (4), 4-O-(E)-caffeoylquinic acid (5), 5-O-(E)-caffeoylquinic acid (6), 3,5-di-O-(E)-caffeoylquinic acid (7), 4,5-di-O-(E)-caffeoylquinic acid (8), (-)-2-O-(E)-caffeoyl-L-threonic acid (9), (-)-4-O-(E)-caffeoyl-L-threonic acid (10), and (-)-4-O-(E)-p-coumaroyl-L-threonic acid (11). Further, (-)-2-O-(E)-caffeoyl-D-malic acid (12) was isolated from C. submollis and also identified for C. pentagyna and C. nigra by co-chromatography. The isolates 10 and 11 were not found in the authentic fresh specimen, indicating that they may be formed during extraction by acyl migration from the 2-O-acylderivatives. Also, 9 and 11 are described here for the first time. All structures were assigned on the basis of their spectroscopic data (1H-, 13C-NMR, MS, optical rotation). PMID:26171328

  4. Analysis of fatty acids and phytosterols in ethanol extracts of Nelumbo nucifera seeds and rhizomes by GC-MS.

    PubMed

    Zhao, Xu; Shen, Jian; Chang, Kyung Ja; Kim, Sung Hoon

    2013-07-17

    The purpose of this study was to investigate the fatty acid and phytosterol contents in ethanol extracts of lotus seeds and rhizomes. These ethanol extracts were extracted with hexane. The hexane extracts were hydrolyzed in a microwave reactor, and total fatty acids and phytosterols were analyzed. The hexane extracts were also subjected to silica gel column chromatography. Nonpolar components (triglycerides and steryl-fatty acid esters) were hydrolyzed, and then the contents were analyzed. Polar components (diglycerides, monoglycerides, fatty acids, and phytosterols) were analyzed directly. Seeds contained higher concentrations of fatty acids and phytosterols compared to rhizomes. Linoleic acid, palmitic acid, and oleic acid were the main fatty acid components in seeds and rhizomes, and most of them in seeds were in the ester form. In seeds, phytosterols existed mainly in the free form rather than in steryl-fatty acid ester form. β-Sitosterol was the most abundant phytosterol in seeds and rhizomes.

  5. Effects of ultrahigh pressure extraction on yield and antioxidant activity of chlorogenic acid and cynaroside extracted from flower buds of Lonicera japonica.

    PubMed

    Hu, Wen; Guo, Ting; Jiang, Wen-Jun; Dong, Guang-Li; Chen, Da-Wei; Yang, Shi-Lin; Li, He-Ran

    2015-06-01

    The present study was designed to establish and optimize a new method for extracting chlorogenic acid and cynaroside from Lonicera japonica Thunb. through orthogonal experimental designl. A new ultrahigh pressure extraction (UPE) technology was applied to extract chlorogenic acid and cynaroside from L. japonica. The influential factors, including solvent type, ethanol concentration, extraction pressure, time, and temperature, and the solid/liquid ratio, have been studied to optimize the extraction process. The optimal conditions for the UPE were developed by quantitative analysis of the extraction products by HPLC-DAD in comparison with standard samples. In addition, the microstructures of the medicinal materials before and after extraction were studied by scanning electron microscopy (SEM). Furthermore, the extraction efficiency of different extraction methods and the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities of the extracts were investigated. The optimal conditions for extracting chlorogenic acid and cynaroside were as follows: ethanol concentration, 60%; extraction pressure, 400 MPa; extraction time, 2 min; extraction temperature, 30 °C; and the solid/liquid ratio, 1 : 50. Under these conditions, the yields of chlorogenic acid and cynaroside were raised to 4.863% and 0.080%, respectively. Compared with other extraction methods, such as heat reflux extraction (HRE), ultrasonic extraction (UE), and Sohxlet extraction (SE), the UPE method showed several advantages, including higher extraction yield, shorter extraction time, lower energy consumption, and higher purity of the extracts. This study could help better utilize L. japonica flower buds as a readily accessible source of natural antioxidants in food and pharmaceutical industries.

  6. Lipids and Fatty Acids in Algae: Extraction, Fractionation into Lipid Classes, and Analysis by Gas Chromatography Coupled with Flame Ionization Detector (GC-FID).

    PubMed

    Guihéneuf, Freddy; Schmid, Matthias; Stengel, Dagmar B

    2015-01-01

    Despite the number of biochemical studies exploring algal lipids and fatty acid biosynthesis pathways and profiles, analytical methods used by phycologists for this purpose are often diverse and incompletely described. Potential confusion and potential variability of the results between studies can therefore occur due to change of protocols for lipid extraction and fractionation, as well as fatty acid methyl esters (FAME) preparation before gas chromatography (GC) analyses. Here, we describe a step-by-step procedure for the profiling of neutral and polar lipids using techniques such as solid-liquid extraction (SLE), thin-layer chromatography (TLC), and gas chromatography coupled with flame ionization detector (GC-FID). As an example, in this protocol chapter, analyses of neutral and polar lipids from the marine microalga Pavlova lutheri (an EPA/DHA-rich haptophyte) will be outlined to describe the distribution of fatty acid residues within its major lipid classes. This method has been proven to be a reliable technique to assess changes in lipid and fatty acid profiles in several other microalgal species and seaweeds.

  7. Comparison of classic with novel in situ extraction of soil amino acids from grassland soil

    NASA Astrophysics Data System (ADS)

    Chen, J.; Williams, D. G.

    2012-12-01

    Characterization of organic and inorganic soil nitrogen availability is important for determining ecosystem response to global change, as nitrogen limitation is often a major constraint on ecosystem productivity. Classic methods of soil nitrogen extraction involve field collection of soil samples and disturbance of soil aggregates during processing. A novel method of soil amino acid extraction is described that allows the collection of semi-sterile soil water extracts in situ with minimal disturbance to soils. Comparison of samples collected using this novel method to samples collected in parallel using classic methods developed by Brookes et al. 1985 and Kielland 1994 revealed different detectable amino acid N pools relative to ammonium. Glutamate and arginine comprised the highest amino acid N pools from extracts collected from a semiarid grassland site using this new method of extraction. In contrast, samples collected and extracted using the classic method contained higher relative levels of serine, glycine and glutamate. The amounts of dominant amino acids relative to ammonium were significantly greater using the classic method compared to the new method. These observed higher ratios of amino acids to ammonium are likely the result of additional amino acid inputs by lysis of microorganisms which are not removed when filtering in the classic method. Disturbance associated with classic methods of soil N determination may have led to alterations in the quantity and distribution of ammonium and amino acids in extracts. Minimizing disturbance of soil aggregates when sampling nitrogen pools and selection of an appropriate filter for collecting free amino acids may be important for accurately determining nitrogen availability to plant roots and soil microbes.

  8. Extractive fermentation for butyric acid production from glucose by Clostridium tyrobutyricum.

    PubMed

    Wu, Zetang; Yang, Shang-Tian

    2003-04-05

    A novel extractive fermentation for butyric acid production from glucose, using immobilized cells of Clostridium tyrobutyricum in a fibrous bed bioreactor, was developed by using 10% (v/v) Alamine 336 in oleyl alcohol as the extractant contained in a hollow-fiber membrane extractor for selective removal of butyric acid from the fermentation broth. The extractant was simultaneously regenerated by stripping with NaOH in a second membrane extractor. The fermentation pH was self-regulated by a balance between acid production and removal by extraction, and was kept at approximately pH 5.5 throughout the study. Compared with conventional fermentation, extractive fermentation resulted in a much higher product concentration (>300 g/L) and product purity (91%). It also resulted in higher reactor productivity (7.37 g/L. h) and butyric acid yield (0.45 g/g). Without on-line extraction to remove the acid products, at the optimal pH of 6.0, the final butyric acid concentration was only approximately 43.4 g/L, butyric acid yield was 0.423 g/g, and reactor productivity was 6.77 g/L. h. These values were much lower at pH 5.5: 20.4 g/L, 0.38 g/g, and 5.11 g/L. h, respectively. The improved performance for extractive fermentation can be attributed to the reduced product inhibition by selective removal of butyric acid from the fermentation broth. The solvent was found to be toxic to free cells in suspension, but not harmful to cells immobilized in the fibrous bed. The process was stable and provided consistent long-term performance for the entire 2-week period of study.

  9. Recovery of uranium from phosphoric acid medium by polymeric composite beads encapsulating organophosphorus extractants

    SciTech Connect

    Singh, D.K.; Yadav, K.K.; Varshney, L.; Singh, H.

    2013-07-01

    The present study deals with the preparation and evaluation of the poly-ethersulfone (PES) based composite beads encapsulating synergistic mixture of D2EHPA and Cyanex 923 (at 4:1 mole ratio) for the separation of uranium from phosphoric acid medium. SEM was used for the characterization of the composite materials. Addition of 1% PVA (polyvinyl alcohol) improved the internal morphology and porosity of the beads. Additionally, microscopic examination of the composite bead confirmed central coconut type cavity surrounded by porous polymer layer of the beads through which exchange of metal ions take place. Effect of various experimental variables including aqueous acidity, metal ion concentration in aqueous feed, concentration of organic extractant inside the beads, extractant to polymer ratio, liquid to solid (L/S) ratio and temperature on the extraction of uranium was studied. Increase in acidity (1-6 M), L/S ratio (1- 10), metal ion concentration (0.2-3 g/L U{sub 3}O{sub 8}) and polymer to extractant ratio (1:4 -1:10) led to decrease in extraction of uranium. At 5.5 M (comparable to wet process phosphoric acid concentration) the extraction of uranium was about 85% at L/S ratio 5. Increase in extractant concentration inside the bead resulted in enhanced extraction of metal ion. Increase in temperature in the range of 30 to 50 Celsius degrees increased the extraction, whereas further increase to 70 C degrees led to the decrease in extraction of uranium. Amongst various reagents tested, stripping of uranium was quantitative by 12% Na{sub 2}CO{sub 3} solution. Polymeric beads were found to be stable and reusable up-to 10 cycles of extraction/stripping. (authors)

  10. SETG: Nucleic Acid Extraction and Sequencing for In Situ Life Detection on Mars

    NASA Astrophysics Data System (ADS)

    Mojarro, A.; Hachey, J.; Tani, J.; Smith, A.; Bhattaru, S. A.; Pontefract, A.; Doebler, R.; Brown, M.; Ruvkun, G.; Zuber, M. T.; Carr, C. E.

    2016-10-01

    We are developing an integrated nucleic acid extraction and sequencing instrument: the Search for Extra-Terrestrial Genomes (SETG) for in situ life detection on Mars. Our goals are to identify related or unrelated nucleic acid-based life on Mars.

  11. Rosmarinic acid content in antidiabetic aqueous extract of Ocimum canum Sims grown in Ghana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rosmarinic acid (RA) is an important polyphenol that is found in a variety of herbs including Ocimum canum sims (locally called eme or akokobesa in Ghana). Aqueous extracts from the leaves of O.canum are used as an antidiabetic herbal medicine in Ghana. Interestingly, rosmarinic acid content and p...

  12. Influence of gelatinization on the extraction of phenolic acids from wheat fractions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of gelatinization on the analysis of phenolic acids from wheat bran, whole-wheat, and refined flour samples was investigated using two extraction procedures, namely, ultrasonic (UAE) and microwave (MAE). The total phenolic acid (TPA) concentration quantity in wheat bran (2711-2913 µg/g) w...

  13. Effects of hops (Humulus lupulus L.) extract on volatile fatty acid production by rumen bacteria

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aims: To determine the effects of hops extract, on in vitro volatile fatty acid (VFA) production by bovine rumen microorganisms. Methods and Results: When mixed rumen microbes were suspended in media containing carbohydrates, the initial rates of VFA production were suppressed by beta-acid rich hops...

  14. 21 CFR 173.280 - Solvent extraction process for citric acid.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... conventional Aspergillus niger fermentation liquor may be safely used to produce food-grade citric acid in... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Solvent extraction process for citric acid. 173.280 Section 173.280 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN...

  15. 21 CFR 173.280 - Solvent extraction process for citric acid.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... conventional Aspergillus niger fermentation liquor may be safely used to produce food-grade citric acid in... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Solvent extraction process for citric acid. 173.280 Section 173.280 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND...

  16. 21 CFR 173.280 - Solvent extraction process for citric acid.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... conventional Aspergillus niger fermentation liquor may be safely used to produce food-grade citric acid in... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Solvent extraction process for citric acid. 173.280 Section 173.280 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND...

  17. 21 CFR 173.280 - Solvent extraction process for citric acid.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... conventional Aspergillus niger fermentation liquor may be safely used to produce food-grade citric acid in... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Solvent extraction process for citric acid. 173.280 Section 173.280 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND...

  18. Effect of chlorogenic acid on antioxidant activity of Flos Lonicerae extracts*

    PubMed Central

    Wu, Lan

    2007-01-01

    Flos Lonicerae is a medically useful traditional Chinese medicine herb. However, little is known about the antioxidant properties of Flos Lonicerae extracts. Here the antioxidant capacity of water, methanolic and ethanolic extracts prepared from Flos Lonicerae to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and reduce Fe3+ to Fe2+ is examined. Chlorogenic acid, a major component of Flos Lonicerae, is identified and further purified from 70% ethanolic extract with high performance liquid chromatography (HPLC) and its antioxidant capacity is characterized. The total phenolic compounds and chlorogenic acid contents in Flos Lonicerae are determined. The present results demonstrate that the Flos Lonicerae extracts exhibit antioxidant activity and chlorogenic acid is a major contributor to this activity. PMID:17726749

  19. Downstream extraction process development for recovery of organic acids from a fermentation broth.

    PubMed

    Bekatorou, Argyro; Dima, Agapi; Tsafrakidou, Panagiotia; Boura, Konstantina; Lappa, Katerina; Kandylis, Panagiotis; Pissaridi, Katerina; Kanellaki, Maria; Koutinas, Athanasios A

    2016-11-01

    The present study focused on organic acids (OAs) recovery from an acidogenic fermentation broth, which is the main problem regarding the use of OAs for production of ester-based new generation biofuels or other applications. Specifically, 10 solvents were evaluated for OAs recovery from aqueous media and fermentation broths. The effects of pH, solvent/OAs solution ratios and application of successive extractions were studied. The 1:1 solvent/OAs ratio showed the best recovery rates in most cases. Butyric and isobutyric acids showed the highest recovery rates (80-90%), while lactic, succinic, and acetic acids were poorly recovered (up to 45%). The OAs recovery was significantly improved by successive 10-min extractions. Alcohols presented the best extraction performance. The process using repeated extractions with 3-methyl-1-butanol led to the highest OAs recovery. However, 1-butanol can be considered as the most cost-effective option taking into account its price and availability.

  20. Application of ionic liquids based enzyme-assisted extraction of chlorogenic acid from Eucommia ulmoides leaves.

    PubMed

    Liu, Tingting; Sui, Xiaoyu; Li, Li; Zhang, Jie; Liang, Xin; Li, Wenjing; Zhang, Honglian; Fu, Shuang

    2016-01-15

    A new approach for ionic liquid based enzyme-assisted extraction (ILEAE) of chlorogenic acid (CGA) from Eucommia ulmoides is presented in which enzyme pretreatment was used in ionic liquids aqueous media to enhance extraction yield. For this purpose, the solubility of CGA and the activity of cellulase were investigated in eight 1-alkyl-3-methylimidazolium ionic liquids. Cellulase in 0.5 M [C6mim]Br aqueous solution was found to provide better performance in extraction. The factors of ILEAE procedures including extraction time, extraction phase pH, extraction temperatures and enzyme concentrations were investigated. Moreover, the novel developed approach offered advantages in term of yield and efficiency compared with other conventional extraction techniques. Scanning electronic microscopy of plant samples indicated that cellulase treated cell wall in ionic liquid solution was subjected to extract, which led to more efficient extraction by reducing mass transfer barrier. The proposed ILEAE method would develope a continuous process for enzyme-assisted extraction including enzyme incubation and solvent extraction process. In this research, we propose a novel view for enzyme-assisted extraction of plant active component, besides concentrating on enzyme facilitated cell wall degradation, focusing on improvement of bad permeability of ionic liquids solutions.

  1. High efficiency protocol of DNA extraction from Micromys minutus mandibles from owl pellets: a tool for molecular research of cryptic mammal species.

    PubMed

    Buś, Magdalena M; Zmihorski, Michał; Romanowski, Jerzy; Balčiauskienė, Laima; Cichocki, Jan; Balčiauskas, Linas

    2014-01-01

    Owl pellets have high potential as a source of DNA. However, this noninvasive method of collecting DNA is rarely used, and its methodological aspects are poorly understood. We investigated the methodology for DNA extraction and amplification from owl pellets containing the smallest European rodent-the Harvest mouse Micromys minutus-as an example. We used mandibles identified in owl pellets for mitochondrial and nuclear DNA amplification. For DNA extraction, we tested two commercial protocols and utilized a protocol being a combination of two commercial kits which ensured high efficiency of DNA extraction. Additionally, we recorded that the amount of DNA was five times higher in extracts from teeth as compared to DNA extracts from jawbones derived from the same mandible. The quantity of DNA was significantly positively correlated with biological sample weight; however, the age of the pellet remains had an impact on the level of inhibition. We recorded inhibition in 40 % of mtDNA extracts derived from pellets older than 150 months, whereas in DNA extracts from pellets younger than 80 months, we did not observe a negative impact of inhibition on PCR efficiency. The amplification success rate was 89.9 % for the mitochondrial fragment and 39.4 % in the case of the nuclear fragment. We observed partial degradation of DNA evidenced by the fact that the longest fragments that we were able to amplify in the case of mtDNA were 450 and 200 bp for nuDNA. The study shows that pellets can be considered as a source of DNA and have high potential for molecular research in the case of threatened species and species that are difficult to study using standard field techniques.

  2. Characterization and functional properties of mango peel pectin extracted by ultrasound assisted citric acid.

    PubMed

    Wang, Miaomiao; Huang, Bohui; Fan, Chuanhui; Zhao, Kaili; Hu, Hao; Xu, Xiaoyun; Pan, Siyi; Liu, Fengxia

    2016-10-01

    Pectin was extracted from 'Tainong No. 1' mango peels, using a chelating agent-citric acid as extraction medium by ultrasound-assisted extraction (UAE) and conventional extraction (CE) at temperatures of 20 and 80°C. Chemical structures, rheological and emulsifying properties of mango peel pectins (MPPs) were comparatively studied with laboratory grade citrus pectin (CP). All MPPs exhibited higher protein content (4.74%-5.94%), degree of methoxylation (85.43-88.38%), average molecular weight (Mw, 378.4-2858kDa) than the CP, but lower galacuronic acid content (GalA, 52.21-53.35%). CE or UAE at 80°C resulted in significantly higher pectin yield than those at 20°C, while the extraction time for UAE-80°C (15min) was significantly shorter compared to CE-80°C (2h) with comparable pectin yield. Moreover, MPPs extracted at 80°C were observed with higher GalA and protein content, higher Mw, resulting in higher viscosity, better emulsifying capacity and stability, as compared to those extracted at 20°C and the CP. Therefore, these results suggested that MPPs from 'Tainong No. 1' may become a highly promising pectin with good thickening and emulsifying properties, using ultrasound-assisted citric acid as an efficient and eco-friendly extraction method.

  3. Inhibition effects on fermentation of hardwood extracted hemicelluloses by acetic acid and sodium.

    PubMed

    Walton, Sara; van Heiningen, Adriaan; van Walsum, Peter

    2010-03-01

    Extraction of hemicellulose from hardwood chips prior to pulping is a possible method for producing ethanol and acetic acid in an integrated forest bio-refinery, adding value to wood components normally relegated to boiler fuel. Hemicellulose was extracted from hardwood chips using green liquor, a pulping liquor intermediate consisting of aqueous NaOH, Na(2)CO(3), and Na(2)S, at 160 degrees C, held for 110 min in a 20 L rocking digester. The extracted liquor contained 3.7% solids and had a pH of 5.6. The organic content of the extracts was mainly xylo-oligosaccharides and acetic acid. Because it was dilute, the hemicellulose extract was concentrated by evaporation in a thin film evaporator. Concentrates from the evaporator reached levels of up to 10% solids. Inhibitors such as acetic acid and sodium were also concentrated by this method, presenting a challenge for the fermentation organisms. Fermentation experiments were conducted with Escherichia coli K011. The un-concentrated extract supported approximately 70% conversion of the initial sugars in 14 h. An extract evaporated down to 6% solids was also fermentable while a 10% solids extract was not initially fermentable. Strain conditioning was later found to enable fermentation at this level of concentration. Alternative processing schemes or inhibitor removal prior to fermentation are necessary to produce ethanol economically.

  4. Phenolic acids, antioxidant and antiproliferative activities of Naviglio® extracts from Schizogyne sericea (Asteraceae).

    PubMed

    Caprioli, Giovanni; Iannarelli, Romilde; Sagratini, Gianni; Vittori, Sauro; Zorzetto, Christian; Sánchez-Mateo, Candelaria C; Rabanal, Rosa M; Quassinti, Luana; Bramucci, Massimo; Vitali, Luca A; Petrelli, Dezemona; Lupidi, Giulio; Venditti, Alessandro; Maggi, Filippo

    2017-03-01

    Schizogyne sericea, well-known as 'salado', is a halophytic shrub widespread on coastal rocks of Tenerife (Canary Islands). This plant is used traditionally as analgesic, astringent, anti-inflammatory and vulnerary agent. In the present work, we have analysed the aqueous and ethanolic extracts of S. sericea for the content of phenolic acids by HPLC-DAD. The dynamic solid-liquid Naviglio® extractor was used to extract the flowering aerial parts. Aqueous extracts showed higher levels of phenolics than ethanolic extracts. S. sericea extracts were rich in chlorogenic and isochlorogenic acids. The Naviglio® extracts obtained were assayed for in vitro biological activities, namely antioxidant, antimicrobial and cytotoxicity on tumour cells by DPPH, ABTS, FRAP, agar disc-diffusion and MTT methods, respectively. Results showed that aqueous extracts, being richer in phenolic acids, are endowed with relevant radical scavenging activity (TEAC values in the range 208-960 μmol TE/g) while ethanolic extracts exhibited noteworthy antiproliferative effects on tumour cells.

  5. Production of lactic acid from hemicellulose extracts by Bacillus coagulans MXL-9.

    PubMed

    Walton, Sara L; Bischoff, Kenneth M; van Heiningen, Adriaan R P; van Walsum, G Peter

    2010-08-01

    Bacillus coagulans MXL-9 was found capable of growing on pre-pulping hemicellulose extracts, utilizing all of the principle monosugars found in woody biomass. This organism is a moderate thermophile isolated from compost for its pentose-utilizing capabilities. It was found to have high tolerance for inhibitors such as acetic acid and sodium, which are present in pre-pulping hemicellulose extracts. Fermentation of 20 g/l xylose in the presence of 30 g/l acetic acid required a longer lag phase but overall lactic acid yield was not diminished. Similarly, fermentation of xylose in the presence of 20 g/l sodium increased the lag time but did not affect overall product yield, though 30 g/l sodium proved completely inhibitory. Fermentation of hot water-extracted Siberian larch containing 45 g/l total monosaccharides, mainly galactose and arabinose, produced 33 g/l lactic acid in 60 h and completely consumed all sugars. Small amounts of co-products were formed, including acetic acid, formic acid, and ethanol. Hemicellulose extract formed during autohydrolysis of mixed hardwoods contained mainly xylose and was converted into lactic acid with a 94% yield. Green liquor-extracted hardwood hemicellulose containing 10 g/l acetic acid and 6 g/l sodium was also completely converted into lactic acid at a 72% yield. The Bacillus coagulans MXL-9 strain was found to be well suited to production of lactic acid from lignocellulosic biomass due to its compatibility with conditions favorable to industrial enzymes and its ability to withstand inhibitors while rapidly consuming all pentose and hexose sugars of interest at high product yields.

  6. Multi-response optimization of the extraction and derivatization protocol of selected polar metabolites from apple fruit tissue for GC-MS analysis.

    PubMed

    Bekele, Elias A; Annaratone, Carlo E P; Hertog, Maarten L A T M; Nicolai, Bart M; Geeraerd, Annemie H

    2014-05-08

    Given the complexity of the plant metabolome exhibiting a wide spectrum of physico-chemical properties, finding the best compromise for GC-MS analysis is a challenging exercise. In this study, the GC-MS protocol for extracting and analyzing polar metabolites from apple tissue is optimized. We found pure methanol extraction to be slightly better as compared to the two phase methanol/chloroform/water extraction in terms of introducing less degradation of the extracted metabolites while being comparable in extraction efficiency and repeatability. The methanol extraction was superior to the single phase methanol/chloroform/water extraction in all performance measures. A multi-response optimization based on a desirability function was applied to optimize the derivatization. This procedure allows searching for optimal parameters while simultaneously considering overall detection enhancement of metabolites from various metabolic classes. A short oximation at a high temperature in combination with a low silylation temperature gave results similar to a longer oximation at low temperature in combination with a high silylation temperature. Increasing silylation time from 0.5 h to 2 h resulted in an improvement of the silylation reaction.

  7. SOLVENT EXTRACTION PROCESS FOR THE RECOVERY OF METALS FROM PHOSPHORIC ACID

    DOEpatents

    Bailes, R.H.; Long, R.S.

    1958-11-01

    > A solvent extraction process is presented for recovering metal values including uranium, thorium, and other lanthanide and actinide elements from crude industrial phosphoric acid solutions. The process conslsts of contacting said solution with an immisclble organic solvent extractant containing a diluent and a material selected from the group consisting of mono and di alkyl phosphates, alkyl phosphonates and alkyl phosphites. The uranlum enters the extractant phase and is subsequently recovered by any of the methods known to the art. Recovery is improved if the phosphate solution is treated with a reducing agent such as iron or aluminum powder prior to the extraction step.

  8. Protocol of capillary isoelectric focusing to separate extremely acidic and basic proteins.

    PubMed

    Yang, Chun; Zhang, Weibing; Zhang, Jie; Duan, Jicheng; Zhang, Yukui

    2005-01-01

    A new set-up was constructed for capillary isoelectric focusing (CIEF) involving a sampling capillary as a bypass fixed to the separation capillary. Sample solutions were subjected to a previously established pH gradient from the sample capillary. Besides performing conventional CIEF, the separation of ampholytic compounds with isoelectric points (p/s) beyond the pH gradient was carried out on this system. This method was termed as pH gradient driven electrophoresis (PGDE) and the basic mathematical expressions were derived to express the dynamic fundamentals. Proteins such as lysozyme, cytochrome C, and pepsin with p/s higher than 10 or below 3 were separated in a pH gradient provided by Pharmalyte (pH 3-10). Finally, this protocol convincingly exhibited its potential in the separation of a solution of chicken egg white.

  9. Sequential ultrasound-microwave assisted acid extraction (UMAE) of pectin from pomelo peels.

    PubMed

    Liew, Shan Qin; Ngoh, Gek Cheng; Yusoff, Rozita; Teoh, Wen Hui

    2016-12-01

    This study aims to optimize sequential ultrasound-microwave assisted extraction (UMAE) on pomelo peel using citric acid. The effects of pH, sonication time, microwave power and irradiation time on the yield and the degree of esterification (DE) of pectin were investigated. Under optimized conditions of pH 1.80, 27.52min sonication followed by 6.40min microwave irradiation at 643.44W, the yield and the DE value of pectin obtained was respectively at 38.00% and 56.88%. Based upon optimized UMAE condition, the pectin from microwave-ultrasound assisted extraction (MUAE), ultrasound assisted extraction (UAE) and microwave assisted extraction (MAE) were studied. The yield of pectin adopting the UMAE was higher than all other techniques in the order of UMAE>MUAE>MAE>UAE. The pectin's galacturonic acid content obtained from combined extraction technique is higher than that obtained from sole extraction technique and the pectin gel produced from various techniques exhibited a pseudoplastic behaviour. The morphological structures of pectin extracted from MUAE and MAE closely resemble each other. The extracted pectin from UMAE with smaller and more regular surface differs greatly from that of UAE. This has substantiated the highest pectin yield of 36.33% from UMAE and further signified their compatibility and potentiality in pectin extraction.

  10. Application of a continuous distribution model for proton binding by humic acids extracted from acidic lake sediments

    SciTech Connect

    Rhea, J.R.; Young, T.C. )

    1987-01-01

    The proton binding characteristics of humic acids extracted from the sediments of Cranberry Pond, an acidic water body located in the Adirondack Mountain region of New York State, were explored by the application of a nultiligand distribution model. The model characterizes a class of proton binding sites by mean log K values and the standard deviations of log K values and the mean. Mean log K values and their relative abundances were determined directly from experimental titration data. The model accurately predicts the binding of protons by the humic acids for pH values in the range 3.5 to 10.0.

  11. Application of a continuous distribution model for proton binding by humic acids extracted from acidic lake sediments

    NASA Astrophysics Data System (ADS)

    Rhea, James R.; Young, Thomas C.

    1987-10-01

    The proton binding characteristics of humic acids extracted from the sediments of Cranberry Pond, an acidic water body located in the Adirondack Mountain region of New York State, were explored by the application of a multiligand distribution model. The model characterizes a class of proton binding sites by mean log K values and the standard deviations of log K values about the mean. Mean log K values and their relative abundances were determined directly from experimental titration data. The model accurately predicts the binding of protons by the humic acids for pH values in the range 3.5 to 10.0.

  12. 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…

  13. Extractability of elements in sugar maple xylem along a gradient of soil acidity.

    PubMed

    Bilodeau Gauthier, Simon; Houle, Daniel; Gagnon, Christian; Côté, Benoît; Messier, Christian

    2008-01-01

    Dendrochemistry has been used for the historical dating of pollution. Its reliability is questionable due primarily to the radial mobility of elements in sapwood. In the present study, the extractability of seven elements was characterized to assess their suitability for the monitoring of environmental conditions. Nine mature sugar maple trees (Acer saccharum Marsh.), a wide-ranging species in eastern North America that has suffered decline in past decades, were sampled in three Quebec watersheds along a soil acidity gradient. Five-year groups of annual tree rings were treated by sequential chemical extractions using extractants of varying strength (deionized H2O, 0.05 M HCl, and concentrated HNO(3)) to selectively solubilize the elements into three fractions (water-soluble, acid-soluble, and residual). Monovalent K; divalent Ba, Ca, Cd, Mg, Mn; and trivalent Al cations were found mostly in the water-soluble, acid-soluble, and residual fractions, respectively. Forms more likely to be mobile within the tree (water-soluble and acid-soluble) do not seem to be suitable for temporal monitoring because of potential lateral redistribution in sapwood rings. However, certain elements (Cd, Mn) were responsive to current soil acidity and could be used in spatial variation monitoring. Extractability of Al varied according to soil acidity; at less acidic sites, up to 90% of Al was contained in the residual form, whereas on very acidic soils, as much as 45% was found in the water-soluble and acid-soluble fractions. Sequential extractions can be useful for determining specific forms of metals as key indicators of soil acidification.

  14. Studies of the acidic components of the Colorado Green River formation oil shale-Mass spectrometric identification of the methyl esters of extractable acids.

    NASA Technical Reports Server (NTRS)

    Haug, P.; Schnoes, H. K.; Burlingame, A. L.

    1971-01-01

    Study of solvent extractable acidic constituents of oil shale from the Colorado Green River Formation. Identification of individual components is based on gas chromatographic and mass spectrometric data obtained for their respective methyl esters. Normal acids, isoprenoidal acids, alpha, omega-dicarboxylic acids, mono-alpha-methyl dicarboxylic acids and methyl ketoacids were identified. In addition, the presence of monocyclic, benzoic, phenylalkanoic and naphthyl-carboxylic acids, as well as cycloaromatic acids, is demonstrated by partial identification.

  15. Effects of raspberry fruit extracts and ellagic acid on respiratory burst in murine macrophages.

    PubMed

    Raudone, Lina; Bobinaite, Ramune; Janulis, Valdimaras; Viskelis, Pranas; Trumbeckaite, Sonata

    2014-06-01

    The mechanism of action of polyphenolic compounds is attributed to their antioxidant, anti-inflammatory, and anti-proliferative properties and their effects on subcellular signal transduction, cell cycle impairment and apoptosis. A raspberry (Rubus idaeus L.) fruit extract contains various antioxidant active compounds, particularly ellagic acid (EA); however the exact intracellular mechanism of their action is not fully understood. The aim of the study was to evaluate the antioxidant effect of raspberry extracts, and that of ellagic acid by assessment of the production of the reactive oxygen species (ROS) by murine macrophage J774 cells. Raspberry extracts and their active compound EA did not affect or had very minor effects on cell viability. No significant difference in the ROS generation in arachidonic acid stimulated macrophages was determined for raspberry extracts and EA whereas in the phorbol-12 myristate-13 acetate model ROS generation was significantly (p < 0.05) reduced. Our observation that raspberry pomace extracts in vitro reduce ROS production in a J774 macrophage culture suggests that raspberry extract and ellagic acid mediated antioxidant effects may be due to the regulation of NADPH oxidase activity.

  16. Comparison of manual and automated nucleic acid extraction from whole-blood samples.

    PubMed

    Riemann, Kathrin; Adamzik, Michael; Frauenrath, Stefan; Egensperger, Rupert; Schmid, Kurt W; Brockmeyer, Norbert H; Siffert, Winfried

    2007-01-01

    Nucleic acid extraction and purification from whole blood is a routine application in many laboratories. Automation of this procedure promises standardized sample treatment, a low error rate, and avoidance of contamination. The performance of the BioRobot M48 (Qiagen) and the manual QIAmp DNA Blood Mini Kit (Qiagen) was compared for the extraction of DNA from whole blood. The concentration and purity of the extracted DNAs were determined by spectrophotometry. Analytical sensitivity was assessed by common PCR and genotyping techniques. The quantity and quality of the generated DNAs were slightly higher using the manual extraction method. The results of downstream applications were comparable to each other. Amplification of high-molecular-weight PCR fragments, genotyping by restriction digest, and pyrosequencing were successful for all samples. No cross-contamination could be detected. While automated DNA extraction requires significantly less hands-on time, it is slightly more expensive than the manual extraction method.

  17. Mercury analysis of various types of coal using acid extraction and pyrolysis methods

    SciTech Connect

    Jae Young Park; Jong Hyun Won; Tai Gyu Lee

    2006-12-15

    The mercury contents of various types of coal currently consumed in Korea were analyzed using acid extraction and pyrolysis methods. The results of analysis by acid extraction and pyrolysis methods were compared and discussed. Generally, high mercury concentrations of 105.6 to 434.5 ng/g (by acid extraction) and 125.7 to 475.4 ng/g (by pyrolysis) were obtained for tested anthracite coals in this study. For bituminous coals, the mercury contents were 11.5-48 ng/g (by acid extraction) and 12.5-52.4 ng/g (by pyrolysis). For coal samples, much simpler and far less time-consuming pyrolysis method tends to give higher values for the Hg concentration than the acid extraction method (by less than 10%) because of the interference from a UV absorption by SOx generated during thermal destruction of coal matrix. Also, further analysis shows that coals with higher densities have higher mercury contents and that the sulfur and mercury contents of coals are positively correlated with each other. 10 refs., 4 figs., 2 tabs.

  18. Determination of volatile fatty acids in wastewater by solvent extraction and gas chromatography

    NASA Astrophysics Data System (ADS)

    Mkhize, Nontando T.; Msagati, Titus A. M.; Mamba, Bhekie B.; Momba, Maggy

    The purpose of this study was to develop a liquid-liquid extraction method for the analysis of volatile fatty acids collected at the elutriation units of Unit 3, 4 and 5 at Johannesburg Water-Northern Works Wastewater Treatment Plant. Liquid-liquid extraction (LLE) method employing dichloromethane (DCM) and methyl-tert-butyl-ether (MTBE) as extracting solvents was used during the quantitative analysis of volatile fatty acids namely acetic, propionic, butyric, isobutyric, valeric, isovaleric and heptanoic acid. The detection of the extracts was by gas chromatography coupled to a mass spectrometer operating under electron ionization mode (GC-EI-MS). The results showed that MTBE was a better extraction solvent than DCM as it gave much higher recoveries (>5 folds). On the other hand, the overall reactor performance for all the three units in the period when the samples were collected, which was measured by the ratio of propionic to acetic acid was good since the ratio o did not exceed 1.4 with the exception of the samples collected on the 3rd of October where the ratio exceeded 1.4 significantly. The concentration of acetic acid, another indicator for the reactor performance in all three units was way below 800 mg/L thus the digester balance was on par.

  19. Direct lactic acid fermentation of Jerusalem artichoke tuber extract using Lactobacillus paracasei without acidic or enzymatic inulin hydrolysis.

    PubMed

    Choi, Hwa-Young; Ryu, Hee-Kyoung; Park, Kyung-Min; Lee, Eun Gyo; Lee, Hongweon; Kim, Seon-Won; Choi, Eui-Sung

    2012-06-01

    Lactic acid fermentation of Jerusalem artichoke tuber was performed with strains of Lactobacillus paracasei without acidic or enzymatic inulin hydrolysis prior to fermentation. Some strains of L. paracasei, notably KCTC13090 and KCTC13169, could ferment hot-water extract of Jerusalem artichoke tuber more efficiently compared with other Lactobacillus spp. such as L. casei type strain KCTC3109. The L. paracasei strains could utilize almost completely the fructo-oligosaccharides present in Jerusalem artichoke. Inulin-fermenting L. paracasei strains produced c.a. six times more lactic acid compared with L. casei KCTC3109. Direct lactic fermentation of Jerusalem artichoke tuber extract at 111.6g/L of sugar content with a supplement of 5 g/L of yeast extract by L. paracasei KCTC13169 in a 5L jar fermentor produced 92.5 ce:hsp sp="0.25"/>g/L of lactic acid with 16.8 g/L fructose equivalent remained unutilized in 72 h. The conversion efficiency of inulin-type sugars to lactic acid was 98% of the theoretical yield.

  20. Reactive Extraction of Lactic Acid by Using Tri-n-octylamine: Structure of the Ionic Phase.

    PubMed

    Aimer, Matthias; Klemm, Elias; Langanke, Bernd; Gehrke, Helmut; Stubenrauch, Cosima

    2016-03-01

    Lactic acid is a promising biogenic platform chemical which can be produced by fermentation of cellulose and hemicellulose. However, separating lactic acid from the fermentation broth is extremely costly and technically complex. We therefore investigated whether liquid/liquid extraction of lactic acid with tri-n-octylamine is a cost-effective alternative to the existing downstream processing method. In order to find an answer to this question, the structure of the middle phase of the occurring three-phase region, which is enriched with up to 20 wt. % lactic acid, was explored. The results of our IR, small-angle X-ray scattering and NMR measurements show that this phase is ionic and has a bicontinuous structure. Due to the analogy with bicontinuous microemulsions, it should be possible to further enrich the lactic acid, which could lead to a rethink regarding the design of extraction processes.

  1. The Effect of Nucleic Acid Extraction Platforms and Sample Storage on the Integrity of Viral RNA for Use in Whole Genome Sequencing.

    PubMed

    Lewandowski, Kuiama; Bell, Andrew; Miles, Rory; Carne, Simon; Wooldridge, David; Manso, Carmen; Hennessy, Nicola; Bailey, Daniel; Pullan, Steven T; Gharbia, Saheer; Vipond, Richard

    2017-03-01

    Extraction of viral RNA and the storage of sample material are extremely important factors in the detection and whole genome sequencing (WGS) of viral pathogens. Although PCR-based detection methods focus on small amplicons, viral WGS applications require RNA of high quality and integrity for adequate sequence coverage and depth. This study examined the fitness of one manual and four automated RNA extraction platforms commonly used in diagnostic laboratories for use in metagenomic sequencing, how the practice of storing sample material in Qiagen buffer AVL before extraction affected the integrity of viral RNA and its suitability for use in amplicon-based WGS methods, and how the addition of Triton X-100 to buffer AVL affected the capability of the extraction platforms and the integrity of viral RNA in stored samples. This study found that the EZ1 platform gave the best performance of the automated platforms and gave comparable results to the frequently used manual Qiagen extraction protocol when extracted viral RNA was used in metagenomics sequencing. To maintain high levels of viral RNA integrity suitable for amplicon-based WGS, nucleic acid should be extracted from samples immediately, because even short storage periods in buffer AVL have a severe effect on integrity, and the addition of Triton X-100 had little effect on the quality of viral material for WGS.

  2. [Optimization of extraction technology for salidroside, tyrosol, crenulatin and gallic acid in Rhodiolae Crenulatae Radix et Rhizoma with orthogonal test].

    PubMed

    Luo, Xin; Wang, Xue-jing; Zhao, Yi-wu; Huang, Wen-zhe; Wang, Zhen-zhong; Xiao, Wei

    2015-09-01

    The extracting technology of salidroside, tyrosol, crenulatin and gallic acid from Rhodiolae Crenulatae Radix et Rhizoma was optimized. With extraction rate of salidroside, tyrosol, crenulatin and gallic acid as indexes, orthogonal test was used to evaluate effect of 4 factors on extracting technology, including concentration of solvent, the dosage of solvent, duration of extraction, and frequency of extraction. The results showed that, the best extracting technology was to extract in 70% alcohol with 8 times the weight of herbal medicine for 2 times, with 3 hours once. High extraction rate of salidroside, tyrosol, crenulatin and gallic acid were obtained with the present technology. The extracting technology was stable and feasible with high extraction rate of four compounds from Rhodiolae Crenulatae Radix et Rhizoma, it was suitable for industrial production.

  3. Evaluation of a new protocol for enzymatic dynamic kinetic resolution of 3-hydroxy-3-(aryl)propanoic acids.

    PubMed

    Koszelewski, Dominik; Zysk, Małgorzata; Brodzka, Anna; Żądło, Anna; Paprocki, Daniel; Ostaszewski, Ryszard

    2015-12-07

    The application of tandem metal-enzyme dynamic kinetic resolution (DKR) is a powerful tool for the manufacture of high-value chemical commodities. A new protocol of kinetic resolution based on irreversible enzymatic esterification of carboxylic acids with orthoesters was introduced to obtain optically active β-hydroxy esters. This procedure was combined with metal catalyzed racemization of the target substrate providing both (R) and (S) enantiomers of ethyl 3-hydroxy-3-(4-nitrophenyl)propanoate with a high yield of 89% at 40 °C. A substantial influence of the enzyme type, organic co-solvent, and metal catalyst on the conversion and enantioselectivity of the enzymatic dynamic kinetic resolution was noted.

  4. Microbial process for the preparation of acetic acid, as well as solvent for its extraction from the fermentation broth

    DOEpatents

    Gaddy, James L.; Clausen, Edgar C.; Ko, Ching-Whan; Wade, Leslie E.; Wikstrom, Carl V.

    2004-06-22

    A modified water-immiscible solvent useful in the extraction of acetic acid from aqueous streams is a substantially pure mixture of isomers of highly branched di-alkyl amines. Solvent mixtures formed of such a modified solvent with a desired co-solvent, preferably a low boiling hydrocarbon, are useful in the extraction of acetic acid from aqueous gaseous streams. An anaerobic microbial fermentation process for the production of acetic acid employs such solvents, under conditions which limit amide formation by the solvent and thus increase the efficiency of acetic acid recovery. Methods for the direct extraction of acetic acid and the extractive fermentation of acetic acid also employ the modified solvents and increase efficiency of acetic acid production. Such increases in efficiency are also obtained where the energy source for the microbial fermentation contains carbon dioxide and the method includes a carbon dioxide stripping step prior to extraction of acetic acid in solvent.

  5. Microbial process for the preparation of acetic acid, as well as solvent for its extraction from the fermentation broth

    DOEpatents

    Gaddy, James L.; Clausen, Edgar C.; Ko, Ching-Whan; Wade, Leslie E.; Wikstrom, Carl V.

    2007-03-27

    A modified water-immiscible solvent useful in the extraction of acetic acid from aqueous streams is a substantially pure mixture of isomers of highly branched di-alkyl amines. Solvent mixtures formed of such a modified solvent with a desired co-solvent, preferably a low boiling hydrocarbon, are useful in the extraction of acetic acid from aqueous gaseous streams. An anaerobic microbial fermentation process for the production of acetic acid employs such solvents, under conditions which limit amide formation by the solvent and thus increase the efficiency of acetic acid recovery. Methods for the direct extraction of acetic acid and the extractive fermentation of acetic acid also employ the modified solvents and increase efficiency of acetic acid production. Such increases in efficiency are also obtained where the energy source for the microbial fermentation contains carbon dioxide and the method includes a carbon dioxide stripping step prior to extraction of acetic acid in solvent.

  6. [Anti-inflammatory effect of Urtica dioica folia extract in comparison to caffeic malic acid].

    PubMed

    Obertreis, B; Giller, K; Teucher, T; Behnke, B; Schmitz, H

    1996-01-01

    Urtica dioica extract is a traditionary used adjuvant therapeutic in rheumatoid arthritis. The antiphlogistic effects of the urtica dioica folia extract IDS 23 (Extractum Urticae dioicae foliorum) and the main phenolic ingredient caffeic malic acid were tested concerning the inhibitory potential on biosynthesis of arachidonic acid metabolites in vitro. The caffeic malic acid was isolated from Urtica folia extract using gel exclusion- and high performance liquid chromatography and identified by mass spectroscopy and nuclear magnetic resonance. Concerning the 5-lipoxygenase products IDS 23 showed a partial inhibitory effect. The isolated phenolic acid inhibited the synthesis of the leukotriene B4 in a concentration dependent manner. The concentration for halfmaximal inhibition (IC50) was 83 microns/ml in the used assay. IDS 23 showed a strong concentration dependent inhibition of the synthesis of cyclooxygenase derived reactions. The IC50 were 92 micrograms/ml for IDS 23 and 38 micrograms/ml for the caffeic malic acid. Calculating the content in IDS 23 the caffeic malic acid is a possible but not the only active ingredient of the plant extract in the tested assay systems. It is demonstrated that the phenolic component showed a different enzymatic target compared with IDS 23. The antiphlogistic effects observed in vitro may give an explanation for the pharmacological and clinical effects of IDS 23 in therapie of rheumatoid diseases.

  7. Study on synthetic methods of trialkyl phosphate oxide and its extraction behavior of some acids

    SciTech Connect

    Yu, M.J.; Su, Y.F.

    1987-01-01

    Trioctyl phosphine oxide (TOPO) is useful for the extraction of many inorganic and organic compounds. A mixed trialkyl phosphine oxide (TRPO) is similar in property to TOPO. The total number of carbon atoms per molecule of TRPO ranges from 15 to 27. Three methods for synthesizing TRPO are described in this paper. When TRPO is synthesized from an alcohol mixture it is significantly cheaper than a single pure alcohol as required for the production of TOPO; tedious purification steps are eliminated. TRPO is a brown liquid which is very slightly soluble in water. Toxicological measurements of LD50, AMES test, hereditary and accumulative toxicity show that TRPO is safe for use in the extraction of some pharmaceutical and biochemical compounds. Examinations of IR and NMR show that the complex interaction of P=O bond of TRPO with extracted substances is the same as that of TOPO. The distribution coefficients of phosphoric acid, citric acid, malic acid, oxalic acid, and tartaric acid with TRPO are reported. The extraction of these acids is believed to proceed by neutral-complex mechanism.

  8. Recovery and separation of sulfuric acid and iron from dilute acidic sulfate effluent and waste sulfuric acid by solvent extraction and stripping.

    PubMed

    Qifeng, Wei; Xiulian, Ren; Jingjing, Guo; Yongxing, Chen

    2016-03-05

    The recovery and simultaneous separation of sulfuric acid and iron from dilute acidic sulfate effluent (DASE) and waste sulfuric acid (WSA) have been an earnest wish for researchers and the entire sulfate process-based titanium pigment industry. To reduce the pollution of the waste acid and make a comprehensive use of the iron and sulfuric acid in it, a new environmentally friendly recovery and separation process for the DASE and the WSA is proposed. This process is based on the reactive extraction of sulfuric acid and Fe(III) from the DASE. Simultaneously, stripping of Fe(III) is carried out in the loaded organic phase with the WSA. Compared to the conventional ways, this innovative method allows the effective extraction of sulfuric acid and iron from the DASE, and the stripping of Fe(III) from the loaded organic phase with the WSA. Trioctylamine (TOA) and tributyl phosphate (TBP) in kerosene (10-50%) were used as organic phases for solvent extraction. Under the optimal conditions, about 98% of Fe(III) and sulfuric acid were removed from the DASE, and about 99.9% of Fe(III) in the organic phase was stripped with the WSA.

  9. Development of molecularly imprinted poly(methacrylic acid)/silica for clean-up and selective extraction of cholesterol in milk prior to analysis by HPLC-UV.

    PubMed

    Clausen, D N; Visentainer, J V; Tarley, C R T

    2014-10-07

    In the present paper the assessment of a novel molecularly imprinted polymer, poly(methacrylic acid)/silica, for clean-up and selective extraction of cholesterol in milk samples is described. The relative selectivity coefficient (k) values for cholesterol/5-α-cholestane and cholesterol/7-dehydrocholesterol systems were found to be 5.08 and 6.08, respectively, thus attesting the selectivity of the MIP for cholesterol under competitive adsorption with structurally analogous steroid compounds. The milk analysis was initially based on saponification followed by liquid-liquid extraction with n-hexane. Then, the protocol of molecularly imprinted solid phase extraction (MISPE) was carried out by loading the milk hexanic extract through 200 mg of MIP or NIP (non-imprinted polymer) packed into SPE cartridges at a flow rate of 0.6 mL min(-1). The washing step was performed by using n-hexane followed by further elution with ethanol and HPLC-UV analysis at 208 nm. From the breakthrough curve the maximum adsorption capacity of the MIP towards cholesterol was found to be 29.51 mg g(-1). The precision of the MISPE protocol was assessed as intra- and inter-days yielding RSD (relative standard deviations) lower than 4.10%. Cleaner HPLC chromatograms were obtained for milk samples submitted to the MISPE protocol in comparison to the solid phase extraction using the NIP or modified octadecyl silica (C18). Recoveries varying from 96.6 up to 102.2% for milk samples spiked with cholesterol were achieved, thus ensuring the accuracy of the proposed method.

  10. Multiresponse optimization of an extraction procedure of carnosol and rosmarinic and carnosic acids from rosemary.

    PubMed

    Oliveira, Gerlon de A R; de Oliveira, Anselmo E; da Conceição, Edemílson C; Leles, Maria I G

    2016-11-15

    A green solvent-based optimization for rosmarinic acid (RA), carnosol (COH), and carnosic acid (CA) extraction, the three main antioxidants from rosemary, was performed. The conventional solid-liquid extraction was optimized using a central composite design (CCD) followed by the desirability approach. In the CCD analysis the quantitative effects of extraction time (4.8-55.2min), liquid-to-solid ratio (4.6-21.4mLg(-1)), and ethanol content (44.8-95.2% v/v) were determined for the extracted amount of antioxidants, their concentrations in the extract, and the extraction yield. Samples were analyzed by HPLC and the antioxidants were identified by comparison with pure standard retention times and UV spectra. The desirability function that simultaneously maximizes the antioxidants extraction and their concentrations in the final product was validated. The extraction using a hydroalcoholic solution 70% v/v, at low liquid-to-solid ratio (5mLg(-1)), and after 55-min yielded an antioxidant recovery rate of 89.8%, and a final product 4.75 times richer in the main antioxidants than the raw material.

  11. Antioxidant activity and sensory analysis of a rosmarinic acid-enriched extract of garden sage (Salvia officinalis)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A novel extract of S. officinalis (garden sage) was prepared using supercritical carbon dioxide (SC-CO2) extraction, followed by a Soxhlet hot water extraction. The resulting extract was enriched in polyphenols, including rosmarinic acid (RA), which has shown promising health benefits in animals. Th...

  12. Nucleic Acids Research Group (NRG): The Importance of DNA Extraction in Metagenomics: The Gatekeeper to Accurate Results!

    PubMed Central

    Carmical, R.; Nadella, V.; Herbert, Z.; Beckloff, N.; Chittur, S.; Rosato, C.; Perera, A.; Auer, H.; Robinson, M.; Tighe, S.; Holbrook, Jennifer

    2013-01-01

    It is well recognized that the field of metagenomics is becoming a critical tool for studying previously unobtainable population dynamics at both an identification of species level and a functional or transcriptional level. Because the power to resolve microbial information is so important for identifying the components in an mixed sample, metagenomics can be used to study nearly any possible environment or system including clinical, environmental, and industrial, to name a few. Clinically, it may be used to determine sub-populations colonizing regions of the body or determining a rare infection to assist in treatment strategies. Environmentally it may be used to identify microbial populations within a soil, water or air sample, or within a bioreactor to characterize a population- based functional process. The possibilities are endless. However, the accuracy of a metagenomics dataset relies on three important “gatekeepers” including 1) The ability to effectively extract all DNA or RNA from every cell within a sample, 2) The reliability of the methods used for deep or high-throughput sequencing, and 3) The software used to analyze the data. Since DNA extraction is the first step in the technical process of metagenomics, the Nucleic Acid Research Group (NARG) conducted a study to evaluate extraction methods using a synthetic microbial sample. The synthetic microbial sample was prepared from 10 known bacteria at specific concentrations and ranging in diversity. Samples were extracted in duplicate using various popular kit based methods as well as several homebrew protocols then analyzed by NextGen sequencing on an Illumina HiSeq. Results of the study include determining the percent recovery of those organisms by comparing to the known quantity in the original synthetic mix.

  13. Comparison of Aromatic Dithiophoshinic and Phosphinic Acid Derivatives for Minor Actinide Extraction

    SciTech Connect

    John R. Klaehn; Dean R. Peterman; Mason K. Harrup; Richard D. Tillotson; Mitchell R. Greenhalgh; Thomas A. Luther; Jack D. Law; Lee M. Daniels

    2008-03-01

    A new extractant for the separation of actinide(III) and lanthanide(III), bis(otrifluoromethylphenyl) phosphinic acid (O-PA) was synthesized. The synthetic route employed mirrors one that was employed to produce the sulfur containing analog bis(otrifluoromethylphenyl) dithiophosphinic acid (S-PA). Multinuclear NMR spectroscopy was used for elementary characterization of the new O-PA derivative. This new O-PA extractant was used to perform Am(III)/Eu(III) separations and the results were directly compared to those obtained in identical separation experiments using S-PA, an extractant that is known to exhibit separation factors of ~100,000 at low pH. The separations data are presented and discussed in terms comparing the nature of the oxygen atom as a donor to that of the sulfur atom in extractants that are otherwise identical.

  14. Comparison of Dithiophosphinic and Phosphinic Acid Derivatives for Minor Actinide Extraction

    SciTech Connect

    Mason K Harrup; Dean R. Peterman; Thomas A. Luther; Mitchell R. Greenhalgh; John R. Klaehn

    2008-03-01

    A new extractant for the separation of actinide(III) and lanthanide(III), bis(otrifluoromethylphenyl) phosphinic acid (O-PA) was synthesized. The synthetic route employed mirrors one that was employed to produce the sulfur containing analog bis(otrifluoromethylphenyl) dithiophosphinic acid (S-PA). Multinuclear NMR spectroscopy was used for elementary characterization of the new O-PA derivative. This new O-PA extractant was used to perform Am(III)/Eu(III) separations and the results were directly compared to those obtained in identical separation experiments using S-PA, an extractant that is known to exhibit separation factors of ~100,000 at low pH. The separations data are presented and discussed in terms comparing the nature of the oxygen atom as a donor to that of the sulfur atom in extractants that are otherwise identical.

  15. Sensitive life detection: extraction of nucleic acids sorbing to Mars analogue minerals

    NASA Astrophysics Data System (ADS)

    Direito, S. O. L.; Marees, A.; Röling, W. F. M.

    2011-10-01

    The main goal of space missions to Mars is to find irrefutable proof of life. Consequently, the development, evaluation and optimization of sensitive extraction and detection methods for biomarkers are of extreme importance. Our aim consisted in the optimization of sensitive extraction techniques for molecules storing hereditary information (nucleic acids such as DNA), since these are common in life forms. However, adsorption of nucleic acids to mineral matrixes and soils can generate low extraction yields. Therefore, a second aim was to determine adsorption and identify 'problematic' Mars analogue minerals. In addition, the development of a method for quantification of DNA recovery by the use of an internal control was proved to be essential, since sensitive extraction needs information on recovery.

  16. Optimization of an Efficient Protein Extraction Protocol Compatible with Two-Dimensional Electrophoresis and Mass Spectrometry from Recalcitrant Phenolic Rich Roots of Chickpea (Cicer arietinum L.)

    PubMed Central

    Chatterjee, Moniya; Gupta, Sumanti; Bhar, Anirban; Das, Sampa

    2012-01-01

    Two-dimensional electrophoresis and mass spectrometry are undoubtedly two essential tools popularly used in proteomic analyses. Utilization of these techniques however largely depends on efficient and optimized sample preparation, regarded as one of the most crucial steps for recovering maximum amount of reliable information. The present study highlights the optimization of an effective and efficient protocol, capable of extraction of root proteins from recalcitrant phenolic rich tissues of chickpea. The widely applicable TCA-acetone and phenol-based methods have been comparatively evaluated, amongst which the latter appeared to be better suited for the sample. The phenol extraction-based method further complemented with sodium dodecyl sulphate (SDS) and pulsatory treatments proved to be the most suitable method represented by greatest spot number, good resolution, and spot intensities. All the randomly selected spots showed successful identification when subjected to further downstream MALDI-TOF and MS/MS analyses. Hence, the information obtained collectively proposes the present protein extraction protocol to be an effective one that could be applicable for recalcitrant leguminous root samples. PMID:23193474

  17. Optimization of an Efficient Protein Extraction Protocol Compatible with Two-Dimensional Electrophoresis and Mass Spectrometry from Recalcitrant Phenolic Rich Roots of Chickpea (Cicer arietinum L.).

    PubMed

    Chatterjee, Moniya; Gupta, Sumanti; Bhar, Anirban; Das, Sampa

    2012-01-01

    Two-dimensional electrophoresis and mass spectrometry are undoubtedly two essential tools popularly used in proteomic analyses. Utilization of these techniques however largely depends on efficient and optimized sample preparation, regarded as one of the most crucial steps for recovering maximum amount of reliable information. The present study highlights the optimization of an effective and efficient protocol, capable of extraction of root proteins from recalcitrant phenolic rich tissues of chickpea. The widely applicable TCA-acetone and phenol-based methods have been comparatively evaluated, amongst which the latter appeared to be better suited for the sample. The phenol extraction-based method further complemented with sodium dodecyl sulphate (SDS) and pulsatory treatments proved to be the most suitable method represented by greatest spot number, good resolution, and spot intensities. All the randomly selected spots showed successful identification when subjected to further downstream MALDI-TOF and MS/MS analyses. Hence, the information obtained collectively proposes the present protein extraction protocol to be an effective one that could be applicable for recalcitrant leguminous root samples.

  18. SIMPLE METHOD FOR THE EXTRACTION OF PHOTOPIGMENTS AND MYCOSPORINE-LIKE AMINO ACIDS (MAAS) FROM SYMBIODINIUM SPP.

    EPA Science Inventory

    Numerous extraction methods have been developed and used in the quantitation of both photopigments and mycosporine amino acids (MAAs) found in Symbiodinium sp. and zooanthellate metazoans. We have development of a simple, mild extraction procedure using methanol, which when coupl...

  19. A novel microalgal lipid extraction method using biodiesel (fatty acid methyl esters) as an extractant.

    PubMed

    Huang, Wen-Can; Park, Chan Woo; Kim, Jong-Duk

    2017-02-01

    Although microalgae are considered promising renewable sources of biodiesel, the high cost of the downstream process is a significant obstacle in large-scale biodiesel production. In this study, a novel approach for microalgal biodiesel production was developed by using the biodiesel as an extractant. First, wet microalgae with 70% water content were incubated with a mixture of biodiesel/methanol and penetration of the mixture through the cell membrane and swelling of the lipids contained in microalgae was confirmed. Significant increases of lipid droplets were observed by confocal microscopy. Second, the swelled lipid droplets in microalgae were squeezed out using mechanical stress across the cell membrane and washed with methanol. The lipid extraction efficiency reached 68%. This process does not require drying of microalgae or solvent recovery, which the most energy-intensive step in solvent-based biodiesel production.

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

    PubMed

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

    2015-06-01

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

  1. Regularities of extracting humic acids from soils using sodium pyrophosphate solutions

    NASA Astrophysics Data System (ADS)

    Bakina, L. G.; Drichko, V. F.; Orlova, N. E.

    2017-02-01

    Regularities of extracting humic acids from different soil types (soddy-podzolic soil, gray forest soil, and all chernozem subtypes) with sodium pyrophosphate solutions at different pH values (from 5 to 13) have been studied. It is found that, regardless of soil type, the process occurs in two stages through the dissociation of carboxylic groups and phenolic hydroxyls, each of which can be described by a logistic function. Parameters of the logistic equations approximating the extraction of humic acids from soils at different pH values are independent of the content and composition of humus in soils. Changes in the optical density of humic acids extracted from soils using sodium pyrophosphate solutions with different pH values are described in the first approximation by the Gaussian function. The optically densest humic acids are extracted using sodium pyrophosphate solutions at pH 10. Therefore, it is proposed to use an extract with pH 10 for the characterization of organic matter with the maximum possible degree of humification in the given soil.

  2. Distribution of zirconium in petroleum sulfoxides during extraction and sorption from nitric and hydrochloric acid solutions

    SciTech Connect

    Turanov, A.N.

    1988-11-20

    Petroleum sulfoxides (PSO) are effective extractants for several metals. We discussed the distribution of petroleum sulfoxides and zirconium between aqueous solutions of hydrochloric and nitric acid and organic solvents, and also the macroporous sorbent impregnated with PSO. For the investigation we used a macroposous copolymer of styrene with divinylbenzene. Our investigation showed a noticeable decrease in the contamination of the raffinates by petroleum sulfoxides and their more complete utilization as extractant of metals from solutions of acids when PSO is deposited on a macroporous copolymer of styrene with divinylbenzene.

  3. Enhanced absorption of boswellic acids by a lecithin delivery form (Phytosome(®)) of Boswellia extract.

    PubMed

    Hüsch, Jan; Bohnet, Janine; Fricker, Gert; Skarke, Carsten; Artaria, Christian; Appendino, Giovanni; Schubert-Zsilavecz, Manfred; Abdel-Tawab, Mona

    2013-01-01

    The anti-inflammatory potential of Boswellia serrata gum resin extracts has been demonstrated in vitro and in animal studies as well as in pilot clinical trials. However, pharmacokinetic studies have evidenced low systemic absorption of boswellic acids (BAs), especially of KBA and AKBA, in rodents and humans. This observation has provided a rationale to improve the formulation of Boswellia extract. We present here the results of a murine comparative bioavailability study of Casperome™, a soy lecithin formulation of standardized B. serrata gum resin extract (BE), and its corresponding non-formulated extract. The concentration of the six major BAs [11-keto-β-boswellic acid (KBA), acetyl-11-keto-β-boswellic acid (AKBA), β-boswellic acid (βBA), acetyl-β-boswellic acid (AβBA), α-boswellic acid (αBA), and acetyl-α-boswellic acid (AαBA)] was evaluated in the plasma and in a series of tissues (brain, muscle, eye, liver and kidney), providing the first data on tissue distribution of BAs. Weight equivalent and equimolar oral administration of Casperome™ provided significantly higher plasma levels (up to 7-fold for KBA, and 3-fold for βBA quantified as area under the plasma concentration time curve, AUC(last)) compared to the non-formulated extract. This was accompanied by remarkably higher tissue levels. Of particular relevance was the marked increase in brain concentration of KBA and AKBA (35-fold) as well as βBA (3-fold) following Casperome™ administration. Notably, up to 17 times higher BA levels were observed in poorly vascularized organs such as the eye. The increased systemic availability of BAs and the improved tissue distribution, qualify Casperome™ for further clinical development to fully exploit the clinical potential of BE.

  4. Water-enhanced solubility of carboxylic acids in organic solvents and its application to extraction processes

    SciTech Connect

    Starr, J.N. ); King, C.J. )

    1992-11-01

    This paper reports on solubilities of carboxylic acids in certain organic solvents which increase sharply as the concentration of water in the solvent increases. This phenomenon leads to a method of regeneration for solvent-extraction processes whereby coextracted water is selectively removed from the extract, such as by stripping, thereby precipitating the acid. The removal of a minor constituent to cause precipitation reduces energy consumption, in contrast with bulk removal of solvent. Solubilities of fumaric acid were measured in a number of organic solvents, with varying amounts of water in the organic phase. Cyclohexanone and methylcyclohexanone were chosen as solvents for which detailed solid-liquid and liquid-liquid equilibria were measured for adipic, fumaric, and succinic acids in the presence of varying concentrations of water, at both 25 and 45[degrees]C. Batch precipitation experiments were performed to demonstrate the processing concept and determine the relative volatility of water to solvent in the presence of carbon.

  5. The removal of uranium from acidic media using ion exchange and/or extraction chromatography

    SciTech Connect

    FitzPatrick, J.R.; Schake, B.S.; Murphy, J.; Holmes, K; West, M.H.

    1996-06-01

    The separation and purification of uranium from either nitric acid or hydrochloric acid media can be accomplished by using either solvent extraction or ion-exchange. Over the past two years at Los Alamos, emerging programs are focused on recapturing the expertise required to do limited, small-quantity processing of enriched uranium. During this period of time, we have been investigating ion-addition, waste stream polishing is associated with this effort in order to achieve more complete removal of uranium prior to recycle of the acid. Extraction chromatography has been demonstrated to further polish the uranium from both nitric and hydrochloric acid media thus allowing for a more complete recovery of the actinide material and creation of less waste during the processing steps.

  6. One-stop genomic DNA extraction by salicylic acid-coated magnetic nanoparticles.

    PubMed

    Zhou, Zhongwu; Kadam, Ulhas S; Irudayaraj, Joseph

    2013-11-15

    Salicylic acid-coated magnetic nanoparticles were prepared via a modified one-step synthesis and used for a one-stop extraction of genomic DNA from mammalian cells. The synthesized magnetic particles were used for magnetic separation of cells from the media by nonspecific binding of the particles as well as extraction of genomic DNA from the lysate. The quantity and quality were confirmed by agarose gel electrophoresis and polymerase chain reaction. The entire process of extraction and isolation can be completed within 30 min. Compared with traditional methods based on centrifugation and filtration, the established method is fast, simple, reliable, and environmentally friendly.

  7. Quinic acid is a biologically active component of the Uncaria tomentosa extract C-Med 100.

    PubMed

    Akesson, Christina; Lindgren, Hanna; Pero, Ronald W; Leanderson, Tomas; Ivars, Fredrik

    2005-01-01

    We have previously reported that the C-Med 100 extract of the plant Uncaria tomentosa induces prolonged lymphocyte half life and hence increased spleen cell number in mice receiving the extract in their drinking water. Further, the extract induces cell proliferation arrest and inhibits activation of the transcriptional regulator nuclear factor kappaB (NF-kappaB) in vitro. We now report that mice exposed to quinic acid (QA), a component of this extract, had significantly increased number of spleen cells, thus recapitulating the in vivo biological effect of C-Med 100 exposure. Commercially supplied QA (H(+) form) did not, however, inhibit cell proliferation in vitro, while the ammonia-treated QA (QAA) was a potent inhibitor. Both QA and QAA inhibited NF-kappaB activity in exposed cells at similar concentrations. Thus, our present data identify QA as a candidate component for both in vivo and in vitro biological effects of the C-Med 100 extract.

  8. Method for extracting lanthanides and actinides from acid solutions by modification of purex solvent

    DOEpatents

    Horwitz, E. Philip; Kalina, Dale G.

    1986-01-01

    A process for the recovery of actinide and lanthanide values from aqueous solutions with an extraction solution containing an organic extractant having the formula: ##STR1## where .phi. is phenyl, R.sup.1 is a straight or branched alkyl or alkoxyalkyl containing from 6 to 12 carbon atoms and R.sup.2 is an alkyl containing from 3 to 6 carbon atoms and phase modifiers in a water-immiscible hydrocarbon diluent. The addition of the extractant to the Purex process extractant, tri-n-butylphosphate in normal paraffin hydrocarbon diluent, will permit the extraction of multivalent lanthanide and actinide values from 0.1 to 12.0 molar acid solutions.

  9. Extraction of lycopene from tomato paste by ursodeoxycholic acid using the selective inclusion complex method.

    PubMed

    Seifi, Mahmoud; Seifi, Parisa; Hadizadeh, Farzin; Mohajeri, Seyed Ahmad

    2013-11-01

    Lycopene, a precursor of β-carotene with well-known antioxidant activity and powerful health properties, can be found in many natural products such as tomato (Lycopersicon esculentum), watermelon, red pepper, and papaya. Many separation methods have been reported for extracting lycopene from its sources. The inclusion complex is an effective method for extraction and purification of organic chemicals. This procedure has 2 main components: host and guest molecules. In this study, lycopene (guest) was extracted from tomato paste by ursodeoxycholic acid, the inclusive agent (host). The molecular structure of the extracted lycopene was then confirmed by (1) HNMR and its purity was evaluated using high-performance liquid chromatography and UV-Vis spectrophotometry methods, in comparison with a standard product. The results indicated that the proposed separation method was very promising and could be used for the extraction and purification of lycopene from tomato paste.

  10. Method for extracting lanthanides and actinides from acid solutions by modification of Purex solvent

    DOEpatents

    Horwitz, E.P.; Kalina, D.G.

    1986-03-04

    A process is described for the recovery of actinide and lanthanide values from aqueous solutions with an extraction solution containing an organic extractant having the formula as shown in a diagram where [phi] is phenyl, R[sup 1] is a straight or branched alkyl or alkoxyalkyl containing from 6 to 12 carbon atoms and R[sup 2] is an alkyl containing from 3 to 6 carbon atoms and phase modifiers in a water-immiscible hydrocarbon diluent. The addition of the extractant to the Purex process extractant, tri-n-butylphosphate in normal paraffin hydrocarbon diluent, will permit the extraction of multivalent lanthanide and actinide values from 0.1 to 12.0 molar acid solutions. 6 figs.

  11. [Rapid determination of eight organic acids in plant tissue by sequential extraction and high performance liquid chromatography].

    PubMed

    Huang, Tianzhi; Wang, Shijie; Liu Xiuming; Liu, Hong; Wu, Yanyou; Luo Xuqiang

    2014-12-01

    A sequential extraction method was developed to determine different forms of oxalate and seven oxalate-metabolism-related organic acids (glyoxylic acid, tartaric acid, glycolic acid, malic acid, acetic acid, citric acid, succinic acid) in plant tissue. The ultra-pure water was used as the extraction medium to obtain water-soluble oxalic acid and the other seven water-soluble organic acids. After the extraction of the water-soluble organic acids, the residues were extracted by dilute hydrochloric acid successively to get the acid-soluble oxalate which entered the liquid phase. A Hypersil ODS column was used with 5 mmol/L potassium dihydrogen phosphate buffer solution (pH 2. 8) as the mobile phase. The diode array detector was set at 210 nm and the column temperature at 30 °C with the injection volume of 5 µL. The flow rate was controlled at different times which allowed a good and rapid separation of the organic acids and hydrochloric acid. Under these conditions, the linear ranges of the method were 1-2000 mg/L for oxalic acid, 25-2,000 mg/L for acetic acid, and 10-2,000 mg/L for glyoxylic acid, tartaric acid, glycolic acid, malic acid, citric acid and succinic acid, with the correlation coefficients of the eight organic acids ≥ 0. 9996. The average recoveries of the eight organic acids in leaves and roots were 93. 5%-104. 4% and 85. 3%-105. 4% with RSDs of 0. 15% -2.43% and 0. 31%-2. 9% (n=7), respectively. The limits of detection ranged from 1 to 10 ng (S/N=3). The results indicated that the method is accurate, rapid and reproducible for the determination of organic acids in plant samples.

  12. Plant uptake of cadmium from acid-extracted anaerobically digested sewage sludge. [Beta vulgaris

    SciTech Connect

    Logan, T.J.; Feltz, R.E.

    1985-01-01

    Approximately 80% of the Cd in an anaerobically digested sewage sludge was removed by acid extraction and dewatering. The acid extracted sludge was treated by (i) neutralization to pH 5.9 with Ca(OH)/sub 2/, (ii) addition of monocalcium phosphate (MCP) followed by Ca(OH)/sub 2/ neutralization to pH 5.9, and, (iii) addition of rock phosphate (RP) followed by Ca(OH)/sub 2/ neutralization to pH 5.9. The three treated sludges and the non acid-extracted sludge were applied to Spinks loamy sand at rates equivalent to 18.7 and 37.4 ..mu..mol Cd kg/sup -1/. Swiss chard (Beta vulgaris) was grown in the greenhouse for 56 d. Cadmium, Fe, Ca, and P were measured in saturation extracts of treated soil after sludge addition. These data indicated that hydroxyapatite was stable throughout the study in the soil receiving MCP treated sludge but not in other soil treatments. Cadmium concentration in saturation extracts of the soil with MCP sludge decreased while Cd concentration in saturation extracts of the other sludge treatments were much higher throughout the study. Chard yields were higher in the control than in any of the sludge treatments, and the difference was attributed to greater N availability in the control. Cadmium concentration in Swiss chard tissue at harvest was significantly lower from the MCP sludge than from the other sludges. Cadmium concentration in chard tissue was also higher from the aerated sludge (11.9 mmol Cd kg/sup -1/) than from the three acid-extracted sludges (2.58-3.29 mmol Cd kg/sup -1/). No significant difference in the Cd concentration of chard was obtained for the 18.7 and 37.4 ..mu..mol Cd kg/sup -1/ rates of the MCP sludge, while Cd concentrations in chard increased linearly with Cd applied by the other sludges.

  13. 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.

  14. Fatty acid profile and elemental content of avocado (Persea americana Mill.) oil--effect of extraction methods.

    PubMed

    Reddy, Mageshni; Moodley, Roshila; Jonnalagadda, Sreekanth B

    2012-01-01

    Interest in vegetable oil extracted from idioblast cells of avocado fruit is growing. In this study, five extraction methods to produce avocado oil have been compared: traditional solvent extraction using a Soxhlet or ultrasound, Soxhlet extraction combined with microwave or ultra-turrax treatment and supercritical fluid extraction (SFE). Traditional Soxhlet extraction produced the most reproducible results, 64.76 ± 0.24 g oil/100 g dry weight (DW) and 63.67 ± 0.20 g oil/100 g DW for Hass and Fuerte varieties, respectively. Microwave extraction gave the highest yield of oil (69.94%) from the Hass variety. Oils from microwave extraction had the highest fatty acid content; oils from SFE had wider range of fatty acids. Oils from Fuerte variety had a higher monounsaturated: saturated FA ratio (3.45-3.70). SFE and microwave extraction produced the best quality oil, better than traditional Soxhlet extraction, with the least amount of oxidizing metals present.

  15. Identification of Bioactivity, Volatile and Fatty Acid Profile in Supercritical Fluid Extracts of Mexican arnica

    PubMed Central

    García-Pérez, J. Saúl; Cuéllar-Bermúdez, Sara P.; Arévalo-Gallegos, Alejandra; Rodríguez-Rodríguez, José; Iqbal, Hafiz M. N.; Parra-Saldivar, Roberto

    2016-01-01

    Supercritical fluid extraction (SFE) is a sustainable technique used for the extraction of lipophilic metabolites such as pigments and fatty acids. Arnica plant is considered a potential candidate material with high antioxidant and antimicrobial activities. Therefore, in this study, a locally available Heterotheca inuloides, also known as Mexican arnica, was analyzed for the extraction of high-value compounds. Based on different pressure (P), temperature (T), and co-solvent (CoS), four treatments (T) were prepared. A maximum 7.13% yield was recovered from T2 (T = 60 °C, P = 10 MPa, CoS = 8 g/min), followed by 6.69% from T4 (T = 60 °C, P = 30 MPa, CoS = 4 g/min). Some bioactive sesquiterpenoids such as 7-hydroxycadalene, caryophyllene and δ-cadinene were identified in the extracts by GC/MS. The fatty acid profile revealed that the main components were palmitic acid (C16:0), followed by linoleic acid (C18:2ω6c), α-linolenic acid (C18:3ω3) and stearic acid (C18:0) differing in percent yield per treatment. Antibacterial activities were determined by the agar diffusion method, indicating that all the treatments exerted strong antibacterial activity against S. aureus, C. albicans, and E. coli strains. The antioxidant capacity of the extracts was also measured by three in vitro assays, DPPH, TEAC and FRAP, using Trolox as a standard. Results showed high antioxidant capacity enabling pharmaceutical applications of Mexican arnica. PMID:27626416

  16. Identification of Bioactivity, Volatile and Fatty Acid Profile in Supercritical Fluid Extracts of Mexican arnica.

    PubMed

    García-Pérez, J Saúl; Cuéllar-Bermúdez, Sara P; Arévalo-Gallegos, Alejandra; Rodríguez-Rodríguez, José; Iqbal, Hafiz M N; Parra-Saldivar, Roberto

    2016-09-12

    Supercritical fluid extraction (SFE) is a sustainable technique used for the extraction of lipophilic metabolites such as pigments and fatty acids. Arnica plant is considered a potential candidate material with high antioxidant and antimicrobial activities. Therefore, in this study, a locally available Heterotheca inuloides, also known as Mexican arnica, was analyzed for the extraction of high-value compounds. Based on different pressure (P), temperature (T), and co-solvent (CoS), four treatments (T) were prepared. A maximum 7.13% yield was recovered from T2 (T = 60 °C, P = 10 MPa, CoS = 8 g/min), followed by 6.69% from T4 (T = 60 °C, P = 30 MPa, CoS = 4 g/min). Some bioactive sesquiterpenoids such as 7-hydroxycadalene, caryophyllene and δ-cadinene were identified in the extracts by GC/MS. The fatty acid profile revealed that the main components were palmitic acid (C16:0), followed by linoleic acid (C18:2ω6c), α-linolenic acid (C18:3ω3) and stearic acid (C18:0) differing in percent yield per treatment. Antibacterial activities were determined by the agar diffusion method, indicating that all the treatments exerted strong antibacterial activity against S. aureus, C. albicans, and E. coli strains. The antioxidant capacity of the extracts was also measured by three in vitro assays, DPPH, TEAC and FRAP, using Trolox as a standard. Results showed high antioxidant capacity enabling pharmaceutical applications of Mexican arnica.

  17. Phytochemicals from Tradescantia albiflora Kunth Extracts Reduce Serum Uric Acid Levels in Oxonate-induced Rats

    PubMed Central

    Wang, Wen-Ling; Sheu, Shi-Yuan; Huang, Wen-Dar; Chuang, Ya-Ling; Tseng, Han-Chun; Hwang, Tzann-Shun; Fu, Yuan-Tsung; Kuo, Yueh-Hsiung; Yao, Chun-Hsu; Kuo, Tzong-Fu

    2016-01-01

    Background: Tradescantia albiflora (TA) Kunth (Commelinaceae) has been used for treating gout and hyperuricemia as folklore remedies in Taiwan. Therefore, it is worthwhile to study the effect of TA extracts on lowering uric acid activity. The hypouricemic effects of TA extracts on potassium oxonate (PO)-induced acute hyperuricemia were investigated for the first time. Materials and Methods: All treatments at the same volume (1 ml) were orally administered to the abdominal cavity of PO-induced hyperuricemic rats. One milliliter of TA extract in n-hexane (HE), ethyl acetate (EA), n-butanol (BuOH), and water fractions has 0.28, 0.21, 0.28, and 1.03 mg TA, respectively; and the plasma uric acid (PUA) level was measured for a consecutive 4 h after administration. Results: All four fractions' extracts derived from TA were observed to significantly reduce PUA compared with the PO group. The EA-soluble fraction (TA-EA) exhibited the best xanthine oxidase (XO) inhibitory activity. Following column chromatography, 12 phytochemicals were isolated and identified from the EA fraction. The IC50 values of isolated phytochemicals indicated that bracteanolide A (AR11) showed the remarkable XO inhibitory effect (IC50 value of 76.4 μg/ml). These findings showed that the in vivo hypouricemic effect in hyperuricemic rats was consistent with in vitro XO inhibitory activity, indicating that TA extracts and derived phytochemicals could be potential candidates as hypouricemic agents. SUMMARY Tradescantia albiflora extracts possess in vivo hypouricemic action in hyperuricemic ratsT. albiflora extracts exhibited strong inhibitory activity against xanthine oxidase (XO)Butenolide may play an important role in XO inhibitionThe extract bracteanolide A was demonstrated potent XO inhibitory activity in vitro. Abbreviations used: TA: Tradescantia albiflora, PO: potassium oxonate, HE: n-hexane, EA: ethyl acetate, BuOH: n-butanol, PUA: plasma uric acid, XO: xanthine oxidase, MeOH: methanol, IP

  18. Simple Protocol for Secondary School Hands-On Activity: Electrophoresis of Pre-Stained Nucleic Acids on Agar-Agar Borate Gels

    ERIC Educational Resources Information Center

    Britos, Leticia; Goyenola, Guillermo; Orono, Silvia Umpierrez

    2004-01-01

    An extremely simple, inexpensive, and safe method is presented, which emulates nucleic acids isolation and electrophoretic analysis as performed in a research environment, in the context of a secondary school hands-on activity. The protocol is amenable to an interdisciplinary approach, taking into consideration the electrical and chemical…

  19. Comparison of the Amino-Acid Content in Pharmacopuncture Extracts Taken from a Scorpion's Body and from Its Tail

    PubMed Central

    Lee, Jin-Ho; Shin, Joon-Shik; Chi, Eun-Hya; Lee, In-Hee

    2013-01-01

    Objective: This study was conducted to investigate the amino-acid compositions of pharmacopuncture extracts taken from the body and from the tail of Buthus martensii Karsch, which are frequently prescribed in Oriental medicine. Methods: Amino acids in hot water and 70% ethanol extracts taken from the scorpion’s whole body and from its tail were screened by using high performance liquid chromatography (HPLC). The experiments were performed with linearity, precision and accuracy. Results: The results of the amino-acid-composition analysis showed that the Buthus martensii Karsch extracts contained various amino acids such as aspartic acid, histidine, alanine, tyrosine, and cystine. The amino-acid analysis showed that the hot water extract was more beneficial than the ethanol extract, except for histidine. The amino acids from the tail and the body of the scorpion were compared, and the concentration of aspartic acid in the extract from the scorpion’s tail was two times that found in the extract from its body. The results of validation experiments were all satisfactory. Conclusion: Studies on the ingredients in extracts from a scorpion other than buthotoxin may demonstrate that the antiepileptic efficacy, anticancer activity, antithrombotic action and analgesic effect are enhanced. Using only the tail of the scorpion when pharmacopuncture is dispensed may be beneficial because the extracts from the tail of the scorpion have higher potency than those from the whole body. PMID:25780666

  20. Extraction of p-coumaric acid and ferulic acid using surfactant-based aqueous two-phase system.

    PubMed

    Dhamole, Pradip B; Demanna, Dhanashree; Desai, S A

    2014-09-01

    Ferulic acid (FA) and p-coumaric acid (pCA) are high-value products that can be obtained by alkaline hydrolysis of lignocellulose. Present work explores the potential of surfactant-based cloud-point extraction (CPE) for FA and pCA extraction from corn cob hydrolysate. More than 90 % (w/w) extraction of both FA and pCA was achieved from model system with L92. The partition coefficient of FA and pCA in L92 aqueous phase system was 35 and 55, respectively. A significant enrichment (8-10-fold) of both FA and pCA was achieved in surfactant-rich phase. Furthermore, the downstream process volume was reduced by 10 to 13 times. Optimized conditions (5 % v/v L92 and pH 3.0) resulted into 85 and 89 % extraction of FA and p-CA, respectively, from alkaline corn cob hydrolysate. Biocompatibility tests were carried out for L92 for ethanol fermentation and found to be biocompatible. Thus, the new surfactant-based CPE system not only concentrated FA and pCA but also reduced the process volume significantly. Further, aqueous phase containing sugars can be used for ethanol fermentation.

  1. Copper-promoted cementation of antimony in hydrochloric acid system: A green protocol.

    PubMed

    Wu, Lian-Kui; Li, Ying-Ying; Cao, Hua-Zhen; Zheng, Guo-Qu

    2015-12-15

    A new method of recovering antimony in hydrochloric acid system by cementation with copper powder was proposed and carried out at laboratory scale. Thermodynamic analysis and cyclic voltammetry test were conducted to study the cementation process. This is a novel antimony removal technology and quite meets the requirements of green chemistry. The main cement product Cu2Sb is a promising anodic material for lithium and sodium ion battery. And nearly all consumed copper powder are transformed into CuCl which is an important industrial material. The effect of reaction temperature, stoichiometric ratio of Cu to Sb(III), stirring rate and concentration of HCl on the cementation efficiency of antimony were investigated in detail. Optimized cementation condition is obtained at 60 °C for 120 min and stirring rate of 600 rpm with Cu/Sb(III) stoichiometric ratio of 6 in 3 mol L(-1) HCl. At this time, nearly all antimony can be removed by copper powder and the cementation efficiency is over 99%. The structure and morphologies of the cement products were characterized by X-ray diffraction and scanning electron microscopy, respectively. Results show that the reaction temperature has little influence on the morphology of the cement products which consist of particles with various sizes. The activation energy of the cementation antimony on copper is 37.75 kJ mol(-1), indicating a chemically controlled step. Inductively coupled plasma mass spectrometry results show that no stibine generates during the cementation process.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  3. Emerging Technology Summary. ACID EXTRACTION TREATMENT SYSTEM FOR TREATMENT OF METAL CONTAMINATED SOILS

    EPA Science Inventory

    The Acid Extraction Treatment System (AETS) is intended to reduce the concentrations and/or teachability of heavy metals in contaminated soils so the soil can be returned to the site from which it originated. The objective of the project was to determine the effectiveness and com...

  4. Rosmarinic acid content in antidiabetic aqueous extract from ocimum canum sims in Ghana

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rosmarinic acid (RA) is an important polyphenol that is found in a variety of herbs including Ocimum canum sims (locally called eme or akokobesa in Ghana). Aqueous extracts from the leaves of O. canum are used as an antidiabetic herbal medicine in Ghana. Analytical TLC was used to examine the compos...

  5. The influence of tributyl phosphate on molybdenum extraction with solutions of dibutyl phosphoric acid

    NASA Astrophysics Data System (ADS)

    Goletskiy, N. D.; Zilberman, B. Ya.; Fedorov, Yu. S.; Khonina, I. V.; Kukharev, D. N.

    2006-01-01

    Comparative investigations were carried out to study the influence of TBP on Mo extraction by HDBP solutions in xylene and TBP in xylene. The dependences of DMo on HNO3 concentration for both HDBP and D2EHPA have minima at about 3 mol/L HNO3. This shows similar extraction properties of HDBP and D2EHPA. The presence of TBP in the solvent results in the reduction of Mo extraction and in an increase in the formal slopes of the falling and rising parts of the logDMo — log[HNO3] curve from -0.5 and +2 up to -2 and +4. Solvent loading curves with Mo show that in the absence of TBP a molybdenum solvate with two molecules of HDBP is formed at low acidity. Anomalous increase in the maximum solvent loading in the presence of TBP is caused by the ability of TBP to extract Mo from oversaturated low acidity solutions following the acidic mechanism. A molybdenum solvate with two HDBP molecules and one HNO3 molecule is possibly formed at high acidity. A flowsheet for Mo recovery from HLW with HDBP-TBP solvent was tested in centrifugal contactors.

  6. Umami taste amino acids produced by hydrolyzing extracted protein from tomato seed meal

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enzymatic hydrolysis was performed for extracting protein to prepare umami taste amino acids from defatted tomato seed meal (DTSM) which is a by-product of tomato processing. Papain was used as an enzyme for the hydrolysis of DTSM. The particle size distribution of DTSM, protein concentration and fr...

  7. TWOPOT: a computer model of the two-pot extractive distillation concept for nitric acid

    SciTech Connect

    Jubin, R.T.; Holland, W.D.; Counce, R.M.; Beckwith, D.R.

    1985-05-01

    A mathematical model, TWOPOT, of the ''two-pot'' extractive distillation concept for nitric acid concentration has been developed. Prediction from a computer simulation using this model shows excellent agreement with the experimental data. This model is recommended for use in the design of large-scale, similar-purpose equipment. 9 refs., 15 figs., 2 tabs.

  8. 21 CFR 173.280 - Solvent extraction process for citric acid.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Solvent extraction process for citric acid. 173.280 Section 173.280 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) SECONDARY DIRECT FOOD ADDITIVES PERMITTED IN FOOD FOR HUMAN CONSUMPTION...

  9. Phenolic acids composition of fruit extracts of Ber (Ziziphus mauritiana L., var. Golo lemai)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fruits of Ziziphus mauritiana L. (ber) are consumed in fresh and dried/processed form in many countries across Asia including Pakistan. In the present study, we analyzed the composition of total phenolic acids (free, soluble-bound and insoluble-bound) from Golo lemai ber fruit extracts by applying a...

  10. Enrichment and separation of chlorogenic acid from the extract of Eupatorium adenophorum Spreng by macroporous resin.

    PubMed

    Liu, Boyan; Dong, Beitao; Yuan, Xiaofan; Kuang, Qirong; Zhao, Qingsheng; Yang, Mei; Liu, Jie; Zhao, Bing

    2016-01-01

    A simple and efficient chromatographic method for separation of chlorogenic acid from Eupatorium adenophorum Spreng extract was developed. The adsorption properties of nine macroporous resins were evaluated. NKA-II resin showed much better adsorption/desorption properties. The adsorption of chlorogenic acid on NKA-II resin at 25°C was well fitted to Langmuir isotherm model and pseudo-second-order kinetic model. The dynamic adsorption and desorption experiments were carried out on columns packed with NKA-II resin to optimize the separation process. The content of chlorogenic acid in the product increased to 22.17%, with a recovery yield of 82.41%.

  11. Lemon juice improves the extractability and quality characteristics of pectin from yellow passion fruit by-product as compared with commercial citric acid extractant.

    PubMed

    Yapo, Beda M

    2009-06-01

    An environment-friendly procedure, allowing the extraction of safe pectin products with good functional properties from yellow passion fruit by-product, was developed using two natural acid extractants, namely, pure lemon juice and citric acid solvent. The results show that both of them solubilise, from cell wall material, pectins characterised by high galacturonic acid content (64-78% w/w), degree of esterification (52-73), viscosity-average molecular weight (70-95 kDa) and capable of forming gels in the presence of high soluble solids (sucrose) content and acid. However, compared to pure citric acid solvents, lemon natural juice and its concentrate isolate, under similar extraction conditions, pectins of superior quality characteristics, i.e., higher galacturonic acid content, degree of esterification, viscosity-average molecular weight and gelling power.

  12. Investigation of metal ion extraction and aggregate formation combining acidic and neutral organophosphorous reagents

    SciTech Connect

    Braatz, A.D.; Nilsson, M.; Ellis, R.; Antonio, M.

    2013-07-01

    In the present study, we investigate how varying mixtures of tri-n-butyl phosphate (TBP) and dibutyl phosphate (HDBP) results in enhanced extraction of lanthanum(III), La{sup 3+}, and dysprosium(III), Dy{sup 3+}. Water and metal ion extraction were carefully monitored as a function of TBP:HDBP mole ratio.In addition to these techniques, EXAFS was used to determine the coordination environment of the metal ion in this system. To produce the necessary signal, a concentration of 1.25*10{sup -3} M La{sup 3+} and Dy{sup 3+} was used. Although previous studies of synergistic extraction of metal cations using combinations of neutral and acidic reagents explain the enhanced extraction by increased dehydration of the metal ion and the formation of mixed extractant complexes, our evidence for the increased water extraction coupled with the aggregate formation suggests a reverse micellar aspect to synergism in the system containing TBP and HDBP. It is quite possible that both of these phenomena contribute to our system behavior. The EXAFS data shows that, based on coordination numbers alone, several possible structures may exist. From this study, we cannot provide a definitive answer as to the nature of extraction in this system or the exact complex formed during extraction.

  13. Evaluation of microwave-assisted extraction for aristolochic acid from Aristolochiae Fructus by chromatographic analysis coupled with nephrotoxicity studies.

    PubMed

    Zhou, Ting; Xiao, Xiao-Hua; Wang, Jia-Yue; Chen, Jin-Ling; Xu, Xian-Fang; He, Zhi-Feng; Li, Gong-Ke

    2012-02-01

    In this paper, a microwave-assisted extraction (MAE) method was established for aristolochic acid-I from Aristolochiae Fructus, and the advantage of MAE was evaluated by chromatographic analysis coupled with nephrotoxicity studies. The experimental parameters of MAE for aristolochic acid-I in Aristolochiae Fructus were investigated and MAE was compared with Soxhlet extraction and ultrasound-assisted extraction in terms of extraction yields and extraction conditions. Under the optimum conditions, MAE could provide higher extraction yields of aristolochic acid-I (1.10 mg/g) than ultrasound-assisted extraction (0.82 mg/g) and Soxhlet extraction (0.95 mg/g), in addition to using less solvent and having a shorter extraction time. Furthermore, the nephrotoxicities of the extracts of Aristolochiae Fructus from different extraction procedures were investigated in Sprague-Dawley rats. The results of nephrotoxicity studies of, for example, general conditions, biochemistry parameters and histopathology examination showed no significantly differences in the nephrotoxicity levels of the extracts from MAE and that from Soxhlet extraction. These results indicated that MAE technique is a simple, rapid and effective extraction method, and the microwave irradiation during MAE procedure did not have any influence on the nephrotoxicity of Aristolochiae Fructus compared with Soxhlet extraction.

  14. Extraction of manganese by alkyl monocarboxylic acid in a mixed extractant from a leaching solution of spent lithium-ion battery ternary cathodic material

    NASA Astrophysics Data System (ADS)

    Joo, Sung-Ho; Shin, Dongju; Oh, ChangHyun; Wang, Jei-Pil; Shin, Shun Myung

    2016-02-01

    We investigate the separation of manganese by an antagonistic effect from a leaching solution of ternary cathodic material of spent lithium-ion batteries that contain 11,400 mg L-1 Co, 11,700 mg L-1 Mn, 12,200 mg L-1 Ni, and 5300 mg L-1 Li using a mixture of alkyl monocarboxylic acid and di-(2-ethylhexyl)phosphoric acid extractants. pH isotherm, distribution coefficient, separation factor, McCabe-Thiele diagram, selective scrubbing, and countercurrent extraction tests are carried out to prove an antagonistic effect and to recover manganese using alkyl monocarboxylic in the mixed extractant. Slope analysis is used to determine the extraction mechanism between a mixture of extractants and valuable metals. An increasing concentration of alkyl monocarboxylic acid in the mixture of extractants results in a decrease in distribution coefficient of cobalt and manganese, however, the separation factor value (β(Mn/Co)) increases at pH 4.5. This is caused by slope analysis where alkyl monocarboxylic acid disrupts the extraction mechanism between di-(2-ethylhexyl)phosphoric acid and cobalt. Finally, continuous countercurrent extraction in a mini-plant test demonstrate the feasibility of manganese recovery from cobalt, nickel, and lithium.

  15. An Advanced TALSPEAK Concept Using 2-Ethylhexylphosphonic Acid Mono-2-Ethylhexyl Ester as the Extractant

    SciTech Connect

    Lumetta, Gregg J.; Casella, Amanda J.; Rapko, Brian M.; Levitskaia, Tatiana G.; Pence, Natasha K.; Carter, Jennifer C.; Niver, Cynthia M.; Smoot, Margaret R.

    2014-12-21

    A method for separating the trivalent actinides and lanthanides is being developed using 2-ethylhexylphosphonic acid mono-2-ethylhexyl ester (HEH[EHP]) as the extractant. The method is based on the preferential binding of the actinides in the aqueous phase by N-(2-hydroxyethyl)ethylenediamine-N,N',N'-triacetic acid (HEDTA), which serves to keep the actinides in the aqueous phase while the lanthanides are extracted into an organic phase containing HEH[EHP]. The process is very robust, showing little dependence upon the pH or the HEH[EHP], HEDTA, and citrate concentrations over the ranges that might be expected in a nuclear fuel recycling plant. Single-stage runs with a 2-cm centrifugal contactor indicate that modifications to the process chemistry may be needed to increase the extraction rate for Sm, Eu, and Gd. The hydraulic properties of the system are favorable to application in centrifugal contactors.

  16. Nucleic acid extraction methods from fixed and paraffin-embedded tissues in cancer diagnostics.

    PubMed

    Bonin, Serena; Stanta, Giorgio

    2013-04-01

    Diagnostic tests, based on nucleic acid extracts from formalin-fixed and paraffin-embedded tissues, are now becoming increasingly common due to the introduction of biological agents for cancer therapy. Unfortunately, the formalin-fixed and paraffin-embedded tissues are heterogeneous in terms of processing and tissue type, and this has an impact on downstream molecular techniques, especially RNA-based techniques. The present review deals with most of the variables connected to the extraction of nucleic acids from formalin-fixed paraffin-embedded tissues, ranging from tissue processing to quality control of extracts. The most recent peer-reviewed publications (mostly published in the past 5 years) and information provided by company websites have been analyzed to compile this review.

  17. Method for the extraction of the volatile compound salicylic acid from tobacco leaf material.

    PubMed

    Verberne, Marianne C; Brouwer, Nynke; Delbianco, Federica; Linthorst, Huub J M; Bol, John F; Verpoorte, Robert

    2002-01-01

    Salicylic acid (SA) is a signalling compound in plants which is able to induce systemic acquired resistance. In the analysis of SA in plant tissues, the extraction recovery is often very low and variable. This is mainly caused by sublimation of SA, especially during evaporation of organic solvents. Techniques have been designed in order to overcome this problem. In the first part of the extraction procedure, sublimation of SA was prevented by addition of 0.2 M sodium hydroxide. At a later stage of the extraction procedure, sublimation of SA during solvent evaporation was controlled by the addition of a small amount of HPLC eluent. In this way, recoveries in the range of 71-91% for free SA and 65-79% for acid-hydrolysed SA were obtained. Recoveries could be further optimised by the use of an internal standard to correct for volume changes after the addition of the HPLC eluent.

  18. Experimental development of a new protocol for extraction and characterization of microplastics in fish tissues: First observations in commercial species from Adriatic Sea.

    PubMed

    Avio, Carlo Giacomo; Gorbi, Stefania; Regoli, Francesco

    2015-10-01

    The presence of microplastics in the marine environment has raised scientific interest during the last decade. Several organisms can ingest microplastics with potentially adverse effects on the digestive tract, respiratory system and locomotory appendages. However, a clear evidence of tissue accumulation and transfer of such microparticles in wild organisms is still lacking, partially hampered by technical difficulties in isolation and characterization protocols from biological samples. In this work, we compared the efficacy of some existing approaches and we optimized a new protocol allowing an extraction yield of microplastics from fish tissues ranging between 78% and 98%, depending on the polymer size. FT-IR analyses confirmed that the extraction procedure did not affect the particles characteristics. The method was further validated on the fish mullet, Mugil cephalus, exposed under laboratory conditions to polystyrene and polyethylene; the particles were isolated and quantified in stomach and liver, and their presence in the hepatic tissue was confirmed also by histological analyses. A preliminary characterization revealed the presence and distribution of microplastics in various fish species collected along the Adriatic Sea. FT-IR analyses indicated polyethylene as the predominant polymer (65%) in the stomach of fish. The overall results confirmed the newly developed method as a reliable approach to detect and quantify microplastics in the marine biota.

  19. Solvent extraction of Sc(III), Zr(IV), Th(IV), Fe(III), and Lu(III) with thiosubstituted organophosphinic acid extractants

    SciTech Connect

    Wang, C.; Li, D.

    1995-05-01

    The solvent extraction of Sc(III), Zr(IV), Th(IV), Fe(III) and Lu(III) with Cyanex 302 (bis(2,4,4-trimethylpentyl)monothiphosphinic acid) and Cyanex 301 (bis(2,4,4-trimethylpentyl)dithiophosphinic acid) in n-hexane from acidic aqueous solutions has been investigated systematically. The effect of equilibrium aqueous acidity on the extraction with these reagents was studied. The separation of Th(IV), Fe(III) and Lu(III) from Sc(III), or the separation of other metals from Lu(III) with Cyanex 302, can be achieved by controlling the aqueous acidity. However, Cyanex 301 exhibited a poor selectivity for the above metals, except for Lu(III). The extraction of these metals with Cyanex 272, Cyanex 302 and Cyanex 301 has been compared. The stripping percentages of Sc(III) for Cyanex 302 and Cyanex 301 in a single stage are near 78% and 75% with 3.5 mol/L and 5.8 mol/L sulphuric acid solutions, respectively. The effects of extractant concentration and temperature on the extraction of Sc(III) were investigated. The stoichiometry of the extraction of Sc(III) with Cyanex 302 was determined. The role of different components of Cyanex 302 in the extraction of Sc(III) was discussed. 18 refs., 10 figs.

  20. Validation of the TaqMan Influenza A Detection Kit and a rapid automated total nucleic acid extraction method to detect influenza A virus in nasopharyngeal specimens.

    PubMed

    Bolotin, Shelly; De Lima, Cedric; Choi, Kam-Wing; Lombos, Ernesto; Burton, Laura; Mazzulli, Tony; Drews, Steven J

    2009-01-01

    This study describes the validation of the TaqMan Influenza A Detection Kit v2.0 combined with an automated nucleic acid extraction method. The limit of detection of this assay was determined by probit regression (95% confidence interval) to be 2 influenza A/PR/8/34 (H1N1) virus particles per microlitre. One hundred and eleven specimens previously tested using the Seeplex RV assay and viral culture methods were tested using the TaqMan Influenza A Detection Kit. Compared to the aggregate gold-standard, the sensitivity and specificity of the TaqMan Influenza A Detection Kit were 100% (35/35) and 97% (74/76), respectively. Because of its accuracy, quick turn-around-time and lyophilized bead form, the TaqMan Influenza A Detection Kit, combined with the NucliSense easyMAG automated extraction method, constitutes a reliable protocol for influenza A diagnosis.

  1. Sensitive life detection strategies for low-biomass environments: optimizing extraction of nucleic acids adsorbing to terrestrial and Mars analogue minerals.

    PubMed

    Direito, Susana O L; Marees, Andries; Röling, Wilfred F M

    2012-07-01

    The adsorption of nucleic acids to mineral matrixes can result in low extraction yields and negatively influences molecular microbial ecology studies, in particular for low-biomass environments on Earth and Mars. We determined the recovery of nucleic acids from a range of minerals relevant to Earth and Mars. Clay minerals, but also other silicates and nonsilicates, showed very low recovery (< 1%). Consequently, optimization of DNA extraction was directed towards clays. The high temperatures and acidic conditions used in some methods to dissolve mineral matrices proved to destruct DNA. The most efficient method comprised a high phosphate solution (P/EtOH; 1 M phosphate, 15% ethanol buffer at pH 8) introduced at the cell-lysing step in DNA extraction, to promote chemical competition with DNA for adsorption sites. This solution increased DNA yield from clay samples spiked with known quantities of cells up to nearly 100-fold. DNA recovery was also enhanced from several mineral samples retrieved from an aquifer, while maintaining reproducible DGGE profiles. DGGE profiles were obtained for a clay sample for which no profile could be generated with the standard DNA isolation protocol. Mineralogy influenced microbial community composition. The method also proved suitable for the recovery of low molecular weight DNA (< 1.5 kb).

  2. A modified acid digestion procedure for extraction of tungsten from soil.

    PubMed

    Bednar, A J; Jones, W T; Chappell, M A; Johnson, D R; Ringelberg, D B

    2010-01-15

    Interest in tungsten occurrence and geochemistry is increasing due to increased use of tungsten compounds and its unknown biochemical effects. Tungsten has a complex geochemistry, existing in most environmental matrices as the soluble and mobile tungstate anion, as well as poly- and heteropolytungstates. Because the geochemistry of tungsten is substantially different than most trace metals, including the formation of insoluble species under acidic conditions, it is not extracted from soil matrices using standard acid digestion procedures. Therefore, the current work describes a modification to a commonly used acid digestion procedure to facilitate quantification of tungsten in soil matrices. Traditional soil digestion procedures, using nitric and hydrochloric acids with hydrogen peroxide yield <1 up to 50% recovery on soil matrix spike samples, whereas the modified method reported here, which includes the addition of phosphoric acid, yields spike recoveries in the 76-98% range. Comparison of the standard and modified digestion procedures on National Institute of Standards and Technology Standard Reference Materials yielded significantly improved tungsten recoveries for the phosphoric acid modified method. The modified method also produces comparable results for other acid extractable metals as the standard methods, and therefore can be used simultaneously for tungsten and other metals of interest.

  3. Topical Formulation Comprising Fatty Acid Extract from Cod Liver Oil: Development, Evaluation and Stability Studies

    PubMed Central

    Ilievska, Biljana; Loftsson, Thorsteinn; Hjalmarsdottir, Martha Asdis; Asgrimsdottir, Gudrun Marta

    2016-01-01

    The purpose of this study was to develop a pharmaceutical formulation containing fatty acid extract rich in free omega-3 fatty acids such as eicosapentaenoic acid and docosahexaenoic acid for topical use. Although the health benefits of cod liver oil and other fish oils taken orally as a dietary supplement have been acknowledged and exploited, it is clear that their use can be extended further to cover their antibacterial properties. In vitro evaluation showed that 20% (v/v) fatty acid extract exhibits good activity against strains of the Gram-positive bacteria Staphylococcus aureus, Enterococcus faecalis, Streptoccoccus pyogenes and Streptoccoccus pneumonia. Therefore, free polyunsaturated fatty acids from cod liver oil or other fish oils can be used as safe and natural antibacterial agents. In this study, ointment compositions containing free fatty acids as active antibacterial agents were prepared by using various natural waxes and characterized. The effects of different waxes, such as carnauba wax, ozokerite wax, laurel wax, beeswax, rice bran wax, candelilla wax and microcrystalline wax, in the concentration range of 1% to 5% (w/w) on the ointment texture, consistency and stability were evaluated. The results showed significant variations in texture, sensory and rheological profiles. This was attributed to the wax’s nature and chain composition. Microcrystalline wax gave the best results but laurel wax, beeswax and rice bran wax exhibited excellent texturing, similar sensory profiles and well-balanced rheological properties. PMID:27258290

  4. Altered nucleic acid partitioning during phenol extraction or silica adsorption by guanidinium and potassium salts.

    PubMed

    Xu, Lei; Lv, Jun; Ling, Liefeng; Wang, Peng; Song, Ping; Su, Ruirui; Zhu, Guoping

    2011-12-15

    Nucleic acids were found to partition into the phenol phase during phenol extraction in the presence of guanidinium at certain concentrations under acidic conditions. The guanidinium-concentration-dependent nucleic acid partitioning patterns were analogous to those of the nucleic acid adsorption/partitioning onto silica mediated by guanidinium, which implied that phenol and silica interact with nucleic acids through similar mechanisms. A competition effect was observed in which the nucleic acids that had partitioned into the phenol phase or onto the silica solid phase could be recovered to the aqueous phases by potassium in a molecular weight-salt concentration-dependent manner (the higher molecular weight nucleic acids needed higher concentrations of potassium to be recovered, and vice versa). Methods were developed based on these findings to isolate total RNA from Escherichia coli. By controlling the concentrations of guanidinium and potassium salts used before phenol extraction or silica adsorption, we can selectively recover total RNA but not the high molecular weight genomic DNA in the aqueous phases. Genomic DNA-free total RNA obtained by our methods is suitable for RT-PCR or other purposes. The methods can also be adapted to isolate small RNAs or RNA in certain molecular weight ranges by changing the salt concentrations used.

  5. Relevance of carnosic acid, carnosol, and rosmarinic acid concentrations in the in vitro antioxidant and antimicrobial activities of Rosmarinus officinalis (L.) methanolic extracts.

    PubMed

    Jordán, Maria J; Lax, Vanesa; Rota, Maria C; Lorán, Susana; Sotomayor, José A

    2012-09-26

    The importance of the diterpenic and rosmarinic acid content in the biological activities of rosemary extracts has been studied previously, but how the relationship between the concentration of these components affects their antioxidant and antibacterial activities has received little attention. Accordingly, from a total of 150 plants, 27 methanolic extracts were selected, for their similar diterpene contents but different ratios between carnosic acid and carnosol concentrations. In extracts with similar rosmarinic acid contents but differing proportions between carnosic acid and carnosol, the two diterpenes were seen to equally affect the in vitro antioxidant activity; however, and related with the antibacterial efficiency, this biological activity improved when carnosol was the major diterpene component.

  6. A Simple and Rapid Protocol for Producing Yeast Extract from Saccharomyces cerevisiae Suitable for Preparing Bacterial Culture Media.

    PubMed

    Zarei, Omid; Dastmalchi, Siavoush; Hamzeh-Mivehroud, Maryam

    2016-01-01

    Yeasts, especially Saccharomyces cerevisiae, are one of the oldest organisms with broad spectrum of applications, owing to their unique genetics and physiology. Yeast extract, i.e. the product of yeast cells, is extensively used as nutritional resource in bacterial culture media. The aim of this study was to develop a simple, rapid and cost benefit process to produce the yeast extract. In this procedure mechanical methods such as high temperature and pressure were utilized to produce the yeast extract. The growth of the bacteria feed with the produced yeast extract was monitored in order to assess the quality of the product. The results showed that the quality of the produced yeast extract was very promising concluded from the growth pattern of bacterial cells in media prepared from this product and was comparable with that of the three commercial yeast extracts in terms of bacterial growth properties. One of the main advantages of the current method was that no chemicals and enzymes were used, leading to the reduced production cost. The method is very simple and cost effective, and can be performed in a reasonable time making it suitable for being adopted by research laboratories. Furthermore, it can be scaled up to produce large quantities for industrial applications.

  7. A Simple and Rapid Protocol for Producing Yeast Extract from Saccharomyces cerevisiae Suitable for Preparing Bacterial Culture Media

    PubMed Central

    Zarei, Omid; Dastmalchi, Siavoush; Hamzeh-Mivehroud, Maryam

    2016-01-01

    Yeasts, especially Saccharomyces cerevisiae, are one of the oldest organisms with broad spectrum of applications, owing to their unique genetics and physiology. Yeast extract, i.e. the product of yeast cells, is extensively used as nutritional resource in bacterial culture media. The aim of this study was to develop a simple, rapid and cost benefit process to produce the yeast extract. In this procedure mechanical methods such as high temperature and pressure were utilized to produce the yeast extract. The growth of the bacteria feed with the produced yeast extract was monitored in order to assess the quality of the product. The results showed that the quality of the produced yeast extract was very promising concluded from the growth pattern of bacterial cells in media prepared from this product and was comparable with that of the three commercial yeast extracts in terms of bacterial growth properties. One of the main advantages of the current method was that no chemicals and enzymes were used, leading to the reduced production cost. The method is very simple and cost effective, and can be performed in a reasonable time making it suitable for being adopted by research laboratories. Furthermore, it can be scaled up to produce large quantities for industrial applications. PMID:28243289

  8. Disposable and removable nucleic acid extraction and purification cartridges for automated flow-through systems

    DOEpatents

    Regan, John Frederick

    2014-09-09

    Removable cartridges are used on automated flow-through systems for the purpose of extracting and purifying genetic material from complex matrices. Different types of cartridges are paired with specific automated protocols to concentrate, extract, and purifying pathogenic or human genetic material. Their flow-through nature allows large quantities sample to be processed. Matrices may be filtered using size exclusion and/or affinity filters to concentrate the pathogen of interest. Lysed material is ultimately passed through a filter to remove the insoluble material before the soluble genetic material is delivered past a silica-like membrane that binds the genetic material, where it is washed, dried, and eluted. Cartridges are inserted into the housing areas of flow-through automated instruments, which are equipped with sensors to ensure proper placement and usage of the cartridges. Properly inserted cartridges create fluid- and air-tight seals with the flow lines of an automated instrument.

  9. DNA extraction protocols cause differences in 16S rRNA amplicon sequencing efficiency but not in community profile composition or structure

    PubMed Central

    Rubin, Benjamin E R; Sanders, Jon G; Hampton-Marcell, Jarrad; Owens, Sarah M; Gilbert, Jack A; Moreau, Corrie S

    2014-01-01

    The recent development of methods applying next-generation sequencing to microbial community characterization has led to the proliferation of these studies in a wide variety of sample types. Yet, variation in the physical properties of environmental samples demands that optimal DNA extraction techniques be explored for each new environment. The microbiota associated with many species of insects offer an extraction challenge as they are frequently surrounded by an armored exoskeleton, inhibiting disruption of the tissues within. In this study, we examine the efficacy of several commonly used protocols for extracting bacterial DNA from ants. While bacterial community composition recovered using Illumina 16S rRNA amplicon sequencing was not detectably biased by any method, the quantity of bacterial DNA varied drastically, reducing the number of samples that could be amplified and sequenced. These results indicate that the concentration necessary for dependable sequencing is around 10,000 copies of target DNA per microliter. Exoskeletal pulverization and tissue digestion increased the reliability of extractions, suggesting that these steps should be included in any study of insect-associated microorganisms that relies on obtaining microbial DNA from intact body segments. Although laboratory and analysis techniques should be standardized across diverse sample types as much as possible, minimal modifications such as these will increase the number of environments in which bacterial communities can be successfully studied. PMID:25257543

  10. DNA extraction protocols cause differences in 16S rRNA amplicon sequencing efficiency but not in community profile composition or structure

    SciTech Connect

    2014-12-01

    The recent development of methods applying next-generation sequencing to microbial community characterization has led to the proliferation of these studies in a wide variety of sample types. Yet, variation in the physical properties of environmental samples demands that optimal DNA extraction techniques be explored for each new environment. The microbiota associated with many species of insects offer an extraction challenge as they are frequently surrounded by an armored exoskeleton, inhibiting disruption of the tissues within. In this study, we examine the efficacy of several commonly used protocols for extracting bacterial DNA from ants. While bacterial community composition recovered using Illumina 16S rRNA amplicon sequencing was not detectably biased by any method, the quantity of bacterial DNA varied drastically, reducing the number of samples that could be amplified and sequenced. These results indicate that the concentration necessary for dependable sequencing is around 10,000 copies of target DNA per microliter. Exoskeletal pulverization and tissue digestion increased the reliability of extractions, suggesting that these steps should be included in any study of insect-associated microorganisms that relies on obtaining microbial DNA from intact body segments. Although laboratory and analysis techniques should be standardized across diverse sample types as much as possible, minimal modifications such as these will increase the number of environments in which bacterial communities can be successfully studied.

  11. DNA extraction protocols cause differences in 16S rRNA amplicon sequencing efficiency but not in community profile composition or structure

    DOE PAGES

    None

    2014-12-01

    The recent development of methods applying next-generation sequencing to microbial community characterization has led to the proliferation of these studies in a wide variety of sample types. Yet, variation in the physical properties of environmental samples demands that optimal DNA extraction techniques be explored for each new environment. The microbiota associated with many species of insects offer an extraction challenge as they are frequently surrounded by an armored exoskeleton, inhibiting disruption of the tissues within. In this study, we examine the efficacy of several commonly used protocols for extracting bacterial DNA from ants. While bacterial community composition recovered using Illuminamore » 16S rRNA amplicon sequencing was not detectably biased by any method, the quantity of bacterial DNA varied drastically, reducing the number of samples that could be amplified and sequenced. These results indicate that the concentration necessary for dependable sequencing is around 10,000 copies of target DNA per microliter. Exoskeletal pulverization and tissue digestion increased the reliability of extractions, suggesting that these steps should be included in any study of insect-associated microorganisms that relies on obtaining microbial DNA from intact body segments. Although laboratory and analysis techniques should be standardized across diverse sample types as much as possible, minimal modifications such as these will increase the number of environments in which bacterial communities can be successfully studied.« less

  12. Extractive and oxidative removal of copper bound to humic acid in soil.

    PubMed

    Hwang, Bo-Ram; Kim, Eun-Jung; Yang, Jung-Seok; Baek, Kitae

    2015-04-01

    Copper (Cu) is often found strongly bound to natural organic matter (NOM) in soil through the formation of strong Cu-NOM complexes. Therefore, in order to successfully remediate Cu-contaminated soils, effective removal of Cu bound to soil organic matter should be considered. In this study, we investigated soil washing methods for Cu removal from a synthetic Cu-contaminated model silica soil coated with humic acid (HA) and from field contaminated soil. Various reagents were studied to extract Cu bound to NOM, which included oxidant (H2O2), base (NaOH), and chelating agents (citric acid and ethylenediaminetetraacetic acid (EDTA)). Among the wash reagents, EDTA extracted Cu most effectively since EDTA formed very strong complexes with Cu, and Cu-HA complexes were transformed into Cu-EDTA complexes. NaOH extracted slightly less Cu compared to EDTA. HA was effectively extracted from the model soil under strongly alkaline conditions with NaOH, which seemed to concurrently release Cu bound to HA. However, chemical oxidation with H2O2 was not effective at destroying Cu-HA complexes. Fourier transform infrared spectroscopy and elemental analysis revealed that chelating agents such as citrate and EDTA were adsorbed onto the model soil via possible complexation between HA and extraction agents. The extraction of Cu from a field contaminated soil sample was effective with chelating agents, while oxidative removal with H2O2 and extractive removal with NaOH separated negligible amounts of Cu from the soil. Based on these results, Cu bound to organic matter in soil could be effectively removed by chelating agents, although remnant agents may remain in the soil.

  13. Should I eXtract Every Six dental trial (SIXES): study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background Extraction of lower first permanent molars in children is common. There is uncertainty among clinicians as to whether a ‘compensating extraction’ (removal of the upper first permanent molar to prevent it over erupting) is necessary despite current guidelines recommending this. As a result, unnecessary dental extractions may be carried out or children may be failing to receive extractions required to achieve optimal long-term oral health. In addition, the decision to extract fewer or more teeth affects management options (local anesthetic injections alone, inhalation sedation or general anesthesia) needed to support the child with the surgical procedure(s). The SIXES (Should I eXtract Every Six) dental trial investigates clinical effectiveness and quality of life for conventional treatment (following the guideline of compensation extraction of the upper first permanent molar) compared with the alternative intervention (removal of lower first permanent molars but no extraction of the upper). Methods/Design This is a multicenter, two-arm parallel group randomized clinical trial. Allocation will be web-based randomization. Practitioners in primary and secondary care settings, reflecting the points of presentation and treatment of eligible patients, will recruit 400 children, aged 7 to 11 years requiring extraction of lower first permanent molars but who have upper first permanent molars of good prognosis. Baseline measures (prior to treatment) and outcome data (at one and five years, or when the patient reaches 14 years of age) will be assessed through study models and child/parent questionnaires. The primary outcome measure is degree of tipping of the lower second permanent molar, (favorable outcome is tipping less than 15°). The secondary outcomes are type of anesthetic/sedation used, residual spacing (between lower second premolar and second permanent molar), orthodontic treatment requirement, quality of life, and over-eruption in the intervention

  14. Selective extraction and determination of chlorogenic acid in fruit juices using hydrophilic magnetic imprinted nanoparticles.

    PubMed

    Hao, Yi; Gao, Ruixia; Liu, Dechun; He, Gaiyan; Tang, Yuhai; Guo, Zengjun

    2016-06-01

    In this paper, the novel hydrophilic magnetic molecularly imprinted nanoparticles were developed for selective separation and determination of chlorogenic acid in aqueous fruit juices. The polymers were prepared by using amino-functionalized magnetic nanoparticles as carriers, branched polyethyleneimine as functional monomer, and chlorogenic acid as template molecule. Branched polyethyleneimine with abundant active amino groups could react with template sufficiently, and its unique dendritic structure may amplify the number of the imprinted cavities. Meanwhile, it would improve the hydrophilicity of imprinted materials for attaining high extraction efficiency. The resulted polymers exhibit fast kinetics, high adsorption capacity, and favorable selectivity. In addition, the obtained nanoparticles were used as solid-phase extraction sorbents for selective isolation and determination of chlorogenic acid in peach, apple, and grape juices (0.92, 4.21, and 0.75 μg mL(-1), respectively).

  15. Extraction-spectrophotometric determination of germanium(IV) with mandelic acid and malachite green.

    PubMed

    Sato, S; Tanaka, H

    1989-03-01

    A method has been developed for determination of germanium, based on complexation with mandelic acid and extraction of the ion-associate formed with Malachite Green (MG) into chlorobenzene. A weakly acidic aqueous solution (pH 2.5-3.5) at room temperature is used and indirect determination is achieved by measuring the absorbance of MG in the extract, at 628 mn. The calibration graph is linear over the range (0.17-8.63) x 10(-6) (0.05-2.50 mug of germanium); the apparent molar absorptivity is 1.33 x 10(5) 1.mole(-1).cm(-1). The interferences from Fe, Ti, Sn(IV), Mo, and SB(III) can be eliminated by addition of trans-1,2-diaminocyclohexanetetra-acetic acid and sodium diethyldithiocarbamate.

  16. Determination of naphthenic acids in crude oils using nonaqueous ion exchange solid-phase extraction.

    PubMed

    Jones, D M; Watson, J S; Meredith, W; Chen, M; Bennett, B

    2001-02-01

    A method is presented for the routine, rapid, and quantitative analysis of aliphatic and naphthenic acids in crude oils, based on their isolation using nonaqueous ion exchange solid-phase extraction cartridges. The isolated acid fractions are methylated and analyzed by gas chromatography and gas chromatography/mass spectrometry. The method is effective on both light and heavy oils and is capable of providing mechanistic information of geochemical significance on the origin of the acids in the oils. Analysis of oils that were solvent extracted from laboratory and field mesocosm marine sediment oil degradation studies indicate that this new method of analyzing the products of hydrocarbon biodegradation may be a useful tool for monitoring the progress of bioremediation of oil spills in the environment.

  17. Liquid extraction of rhenium(VII) and molybdenum(VI) with trialkylphosphine oxide from acidic solutions

    NASA Astrophysics Data System (ADS)

    Travkin, V. F.; Palant, A. A.

    2012-01-01

    The liquid extraction of rhenium(VII) and molybdenum(VI) ions from sulfuric, hydrochloric, and nitric acid media is studied in the temperature range from 20 to 40°C using trialkylphosphine oxide in kerosene as an extracting agent. The maximum separation of these metals is attained when they are extracted from solutions of 1.0-2.0 M H2SO4 (the duration of intense phase mixing was 3-5 min). The enthalpy of the studied process is estimated to be Δ H = -32.32 kJ/mol for molybdenum and -51.52 kJ/mol for rhenium. The chemical aspects of the extraction process studied are discussed.

  18. Comparison of QIAGEN automated nucleic acid extraction methods for CMV quantitative PCR testing.

    PubMed

    Miller, Steve; Seet, Henrietta; Khan, Yasmeen; Wright, Carolyn; Nadarajah, Rohan

    2010-04-01

    We examined the effect of nucleic acid extraction methods on the analytic characteristics of a quantitative polymerase chain reaction (PCR) assay for cytomegalovirus (CMV). Human serum samples were extracted with 2 automated instruments (BioRobot EZ1 and QIAsymphony SP, QIAGEN, Valencia, CA) and CMV PCR results compared with those of pp65 antigenemia testing. Both extraction methods yielded results that were comparably linear and precise, whereas the QIAsymphony SP had a slightly lower limit of detection (1.92 log(10) copies/mL vs 2.26 log(10) copies/mL). In both cases, PCR was more sensitive than CMV antigen detection, detecting CMV viremia in 12% (EZ1) and 21% (QIAsymphony) of antigen-negative specimens. This study demonstrates the feasibility of using 2 different extraction techniques to yield results within 0.5 log(10) copies/mL of the mean value, a level that would allow for clinical comparison between different laboratory assays.

  19. Efficiency and selectivity of triterpene acid extraction from decoctions and tinctures prepared from apple peels

    PubMed Central

    Siani, Antonio C.; Nakamura, Marcos J.; dos Santos, Daniel S.; Mazzei, José L.; do Nascimento, Adriana C.; Valente, Ligia M. M.

    2014-01-01

    Background: This study assessed the extraction efficiency of ursolic (UA) and oleanolic acids (OA), as well as the total phenols in aqueous and hydroethanolic extracts of dry apple peels at room temperature. Materials and Methods: After running preliminary assays on decoctions and tinctures (ethanol: water 7:3 v/v), the extracts from dried apple (cv. Fuji) peels were obtained by static maceration over varied intervals (2 to 180 days). The UA and OA content in the extracts was quantified by High Performance Liquid Chromatography with Diode Array Detection (HPLC-DAD) with a reversed phase column and isocratic elution (CH3CN/H2O/H3PO4) against calibration curves (R2 > 0.9995). The total phenol content in the extracts was evaluated spectrophotometrically at 760 nm using the Folin-Ciocalteau method referencing gallic acid. Results: UA and OA in the hydroethanolic extracts ranged from 3.63-6.12 mg/g and 2.12-3.30 mg/g, corresponding to 1.72-3.07 and 1.00-1.66 mg/g in the raw material, respectively. Higher values of triterpene acid content corresponded to maceration periods of 10 or 30 days. The residual phenol and polyphenol content ranged from 6.97 to 11.6 mg/g. The UA and OA yields, as well as the total phenol content, versus the maceration time were plotted in Control Charts within confidence intervals (95%) and were unaffected during the assayed period. Conclusion: Apple peel tinctures from 10% solids obtained at room temperature exhibited the highest content of triterpene acids when employing a maceration period of 10 to 30 days. Extracts prepared using this procedure contained an average of 7.33 mg/g of total triterpene acids and 10.6 mg/g phenolic compounds. These results establish supporting data for apple peel tinctures and their derived phytopharmaceuticals that are standardized on the ursolic-oleanolic acid content. PMID:24991096

  20. PROCESS FOR EXTRACTING NEPTUNIUM AND PLUTONIUM FROM NITRIC ACID SOLUTIONS OF SAME CONTAINING URANYL NITRATE WITH A TERTIARY AMINE

    DOEpatents

    Sheppard, J.C.

    1962-07-31

    A process of selectively extracting plutonium nitrate and neptunium nitrate with an organic solution of a tertiary amine, away from uranyl nitrate present in an aqueous solution in a maximum concentration of 1M is described. The nitric acid concentration is adjusted to about 4M and nitrous acid is added prior to extraction. (AEC)

  1. Extracts from Tribulus species may modulate platelet adhesion by interfering with arachidonic acid metabolism.

    PubMed

    Olas, Beata; Hamed, Arafa I; Oleszek, Wieslaw; Stochmal, Anna

    2015-01-01

    The present work was designed to study the effects of crude extracts from Tribulus pterocarpus, T. pentandrus and T. parvispinus on selected biological functions of human blood platelets in vitro. Platelet suspensions were pre-incubated with extracts from aerial parts of T. pterocarpus, T. pentandrus and T. parvispinus, at the final concentrations of 0.5, 5 and 50 µg/ml. Then, for platelet activation thrombin, was used. The effects of crude extracts from T. pterocarpus, T. pentandrus and T. parvispinus on adhesion of blood platelets to collagen were determined by method according to Tuszynski and Murphy. Arachidonic acid metabolism was measured by the level of thiobarbituric acid reactive substances (TBARS). In these studies we also compared the action of tested crude plant extracts with the effects of the polyphenolic fraction isolated from aerial parts of T. pterocarpus, which has antiplatelet and antioxidative properties. The performed assays demonstrated that the tested crude extract from T. pterocarpus and the phenolic fraction from T. pterocarpus might influence the platelet functions in vitro. The inhibitory, concentration-dependent effects of this tested extract and its phenolic fraction on adhesion of resting platelets and thrombin - stimulated platelets to collagen was found. We also observed that the crude extract from T. pterocarpus, like the polyphenolic fraction from T. pterocarpus reduced TBARS production in blood platelets. In the comparative studies, the tested crude extract from T. pterocarpus was not found to be more effective antiplatelet factor, than the polyphenolic fraction from this plant. The results obtained suggest that T. pterocarpus may be a promising source of natural compounds, valuable in the prevention of the enhanced activity of blood platelets in numerous cardiovascular diseases.

  2. Anti-leukemic activity of Dillenia indica L. fruit extract and quantification of betulinic acid by HPLC.

    PubMed

    Kumar, Deepak; Mallick, Sumana; Vedasiromoni, Joseph R; Pal, Bikas C

    2010-05-01

    The methanolic extract of Dillenia indica L. fruits showed significant anti-leukemic activity in human leukemic cell lines U937, HL60 and K562. This finding led to fractionation of the methanolic extract, on the basis of polarity, in which the ethyl acetate fraction showed the highest anti-leukemic activity. A major compound, betulinic acid, was isolated from the ethyl acetate fraction by silica gel column chromatography and was identified and characterized. Betulinic acid could explain the anti-leukemic activity of the methanolic extract and the ethyl acetate fraction. Hence the quantitative estimation of betulinic acid was approached in methanolic extract and fractions using HPLC.

  3. A simple, efficient and environmentally benign synthetic protocol for the synthesis of spirooxindoles using choline chloride-oxalic acid eutectic mixture as catalyst/solvent system.

    PubMed

    Khandelwal, Sarita; Rajawat, Anshu; Tailor, Yogesh Kumar; Kumar, Mahendra

    2014-01-01

    An efficient and environmentally benign domino protocol has been presented for the synthesis of structurally diverse spirooxindoles spiroannulated with pyranopyridopyrimidines, indenopyridopyrimidines, and chromenopyridopyrimidines involving three-component reaction of aminouracils, isatins and cyclic carbonyl compounds in deep eutectic solvent (choline chloride-oxalic acid: 1:1) which acts as efficient catalyst and environmentally benign reaction medium. The present protocol offers several advantages such as operational simplicity with easy workup, shorter reaction times excellent yields with superior atom economy and environmentally benign reaction conditions with the use of cost-effective, recyclable, non-toxic and bio-degradable DES as catalyst/solvent.

  4. Extraction, purification, methylation and GC-MS analysis of short-chain carboxylic acids for metabolic flux analysis.

    PubMed

    Tivendale, Nathan D; Jewett, Erin M; Hegeman, Adrian D; Cohen, Jerry D

    2016-08-15

    Dynamic metabolic flux analysis requires efficient and effective methods for extraction, purification and analysis of a plethora of naturally-occurring compounds. One area of metabolism that would be highly informative to study using metabolic flux analysis is the tricarboxylic acid (TCA) cycle, which consists of short-chain carboxylic acids. Here, we describe a newly-developed method for extraction, purification, derivatization and analysis of short-chain carboxylic acids involved in the TCA cycle. The method consists of snap-freezing the plant material, followed by maceration and a 12-15h extraction at -80 °C. The extracts are then subject to reduction (to stabilize β-keto acids), purified by strong anion exchange solid phase extraction and methylated with methanolic HCl. This method could also be readily adapted to quantify many other short-chain carboxylic acids.

  5. Ultrasonic-Assisted Extraction of Raspberry Seed Oil and Evaluation of Its Physicochemical Properties, Fatty Acid Compositions and Antioxidant Activities

    PubMed Central

    Huang, Qun; Wang, Jinli; Lin, Qiyang; Liu, Mingxin; Lee, Won Young; Song, Hongbo

    2016-01-01

    Ultrasonic-assisted extraction was employed for highly efficient separation of aroma oil from raspberry seeds. A central composite design with two variables and five levels was employed and effects of process variables of sonication time and extraction temperature on oil recovery and quality were investigated. Optimal conditions predicted by response surface methodology were sonication time of 37 min and extraction temperature of 54°C. Specifically, ultrasonic-assisted extraction (UAE) was able to provide a higher content of beneficial unsaturated fatty acids, whereas conventional Soxhlet extraction (SE) resulted in a higher amount of saturated fatty acids. Moreover, raspberry seed oil contained abundant amounts of edible linoleic acid and linolenic acid, which suggest raspberry seeds could be valuable edible sources of natural γ-linolenic acid products. In comparison with SE, UAE exerted higher free radical scavenging capacities. In addition, UAE significantly blocked H2O2-induced intracellular reactive oxygen species (ROS) generation. PMID:27120053

  6. Ultrasonic-Assisted Extraction of Raspberry Seed Oil and Evaluation of Its Physicochemical Properties, Fatty Acid Compositions and Antioxidant Activities.

    PubMed

    Teng, Hui; Chen, Lei; Huang, Qun; Wang, Jinli; Lin, Qiyang; Liu, Mingxin; Lee, Won Young; Song, Hongbo

    2016-01-01

    Ultrasonic-assisted extraction was employed for highly efficient separation of aroma oil from raspberry seeds. A central composite design with two variables and five levels was employed and effects of process variables of sonication time and extraction temperature on oil recovery and quality were investigated. Optimal conditions predicted by response surface methodology were sonication time of 37 min and extraction temperature of 54°C. Specifically, ultrasonic-assisted extraction (UAE) was able to provide a higher content of beneficial unsaturated fatty acids, whereas conventional Soxhlet extraction (SE) resulted in a higher amount of saturated fatty acids. Moreover, raspberry seed oil contained abundant amounts of edible linoleic acid and linolenic acid, which suggest raspberry seeds could be valuable edible sources of natural γ-linolenic acid products. In comparison with SE, UAE exerted higher free radical scavenging capacities. In addition, UAE significantly blocked H2O2-induced intracellular reactive oxygen species (ROS) generation.

  7. Temperature shifts for extraction and purification of zygomycetes chitosan with dilute sulfuric acid.

    PubMed

    Zamani, Akram; Edebo, Lars; Niklasson, Claes; Taherzadeh, Mohammad J

    2010-08-13

    The temperature-dependent hydrolysis and solubility of chitosan in sulfuric acid solutions offer the possibility for chitosan extraction from zygomycetes mycelia and separation from other cellular ingredients with high purity and high recovery. In this study, Rhizomucor pusillus biomass was initially extracted with 0.5 M NaOH at 120 °C for 20 min, leaving an alkali insoluble material (AIM) rich in chitosan. Then, the AIM was subjected to two steps treatment with 72 mM sulfuric acid at (i) room temperature for 10 min followed by (ii) 120 °C for 45 min. During the first step, phosphate of the AIM was released into the acid solution and separated from the chitosan-rich residue by centrifugation. In the second step, the residual AIM was re-suspended in fresh 72 mM sulfuric acid, heated at 120 °C and hot filtered, whereby chitosan was extracted and separated from the hot alkali and acid insoluble material (HAAIM). The chitosan was recovered from the acid solution by precipitation at lowered temperature and raised pH to 8-10. The treatment resulted in 0.34 g chitosan and 0.16 g HAAIM from each gram AIM. At the start, the AIM contained at least 17% phosphate, whereas after the purification, the corresponding phosphate content of the obtained chitosan was just 1%. The purity of this chitosan was higher than 83%. The AIM subjected directly to the treatment with hot sulfuric acid (at 120 °C for 45 min) resulted in a chitosan with a phosphate impurity of 18.5%.

  8. Assessment of rosmarinic acid content in six Lamiaceae species extracts and their antioxidant and antimicrobial potential.

    PubMed

    Benedec, Daniela; Hanganu, Daniela; Oniga, Ilioara; Tiperciuc, Brindusa; Olah, Neli-Kinga; Raita, Oana; Bischin, Cristina; Silaghi-Dumitrescu, Radu; Vlase, Laurian

    2015-11-01

    In the present study, six indigenous species of Lamiaceae family (Origanum vulgare L., Melissa officinalis L., Rosmarinus officinalis L., Ocimum basilicum L., Salvia officinalis L. and Hyssopus officinalis L.), have been analyzed to assess the rosmarinic acid, phenyl propane derivatives and polyphenolic contents and their antioxidant and antimicrobial potential. HPLC-MS method has been used for the analysis ofrosmarinicacid. The phenyl propane derivatives and total phenolic contents were determined using spectrophotometric method. The ethanolic extracts were screened for antioxidant activities by DPPH radical scavenging, HAPX (hemoglobin ascorbate per oxidase activity inhibition), and EPR (electron paramagnetic resonance) methods. The ethanolic extracts revealed the presence of rosmarinic acid in the largest amount in O. vulgare (12.40mg/g) and in the lowest in R. officinalis (1.33 mg/g). O. vulgare extracts exhibited the highest antioxidant capacity, in line with the rosmarinic acid and polyphenolic contents. The antimicrobial testing showed a significant activity against L. monocytogenes, S. aureus and C. albicans for all six extracts.

  9. Aqueous extracts of Mozambican plants as alternative and environmentally safe acid-base indicators.

    PubMed

    Macuvele, Domingos Lusitaneo Pier; Sithole, Gerre Zebedias Samo; Cesca, Karina; Macuvele, Suzana Lília Pinare; Matsinhe, Jonas Valente

    2016-06-01

    Indicators are substances that change color as the pH of the medium. Many of these substances are dyes of synthetic origin. The mulala plant (Euclea natalensis), which roots are commonly used by rural communities for their oral hygiene, and roseira (Hibiscus rosa-sinensis), an ornamental plant, are abundant in Mozambique. Currently, synthetic acid-base indicators are most commonly used but have environmental implications and, on the other hand, are expensive products, so the demand for natural indicators started. This study investigated the applicability of aqueous extracts of H. rosa-sinensis and E. natalensis as acid-base indicators. Ground on this work, the extracts can be used as acid-base indicators. On the basis of the absorption spectroscopy in both the UV-Vis region and previous studies, it was possible to preliminarily pinpoint anthocyanins and naphthoquinones as responsible for the shifting of colors depending on the pH range of aqueous extracts of H. rosa-sinensis and E. natalensis. These natural indicators are easily accessible, inexpensive, easy to extract, environmentally safe, and locally available.

  10. Comparison of supercritical fluid extraction and ultrasound-assisted extraction of fatty acids from quince (Cydonia oblonga Miller) seed using response surface methodology and central composite design.

    PubMed

    Daneshvand, Behnaz; Ara, Katayoun Mahdavi; Raofie, Farhad

    2012-08-24

    Fatty acids of Cydonia oblonga Miller cultivated in Iran were obtained by supercritical (carbon dioxide) extraction and ultrasound-assisted extraction methods. The oils were analyzed by capillary gas chromatography using mass spectrometric detections. The compounds were identified according to their retention indices and mass spectra (EI, 70eV). The experimental parameters of SFE such as pressure, temperature, modifier volume, static and dynamic extraction time were optimized using a Central Composite Design (CCD) after a 2(5) factorial design. Pressure and dynamic extraction time had significant effect on the extraction yield, while the other factors (temperature, static extraction time and modifier volume) were not identified as significant factors under the selected conditions. The results of chemometrics analysis showed the highest yield for SFE (24.32%), which was obtained at a pressure of 353bar, temperature of 35°C, modifier (methanol) volume of 150μL, and static and dynamic extraction times of 10 and 60min, respectively. Ultrasound-assisted extraction (UAE) of Fatty acids from C. oblonga Miller was optimized, using a rotatable central composite design. The optimum conditions were as follows: solvent (n-hexane) volume, 22mL; extraction time, 30min; and extraction temperature, 55°C. This resulted in a maximum oil recovery of 19.5%. The extracts with higher yield from both methods were subjected to transesterification and GC-MS analysis. The results show that the oil obtained by SFE with the optimal operating conditions allowed a fatty acid composition similar to the oil obtained by UAE in optimum condition and no significant differences were found. The major components of oil extract were Linoleic, Palmitic, Oleic, Stearic and Eicosanoic acids.

  11. Lipid nutritional value of legumes: Evaluation of different extraction methods and determination of fatty acid composition.

    PubMed

    Caprioli, Giovanni; Giusti, Federica; Ballini, Roberto; Sagratini, Gianni; Vila-Donat, Pilar; Vittori, Sauro; Fiorini, Dennis

    2016-02-01

    This study sought to contribute to the assessment of the nutritional properties of legumes by determining the fatty acid (FA) composition of 29 legume samples after the evaluation of nine extraction methods. The Folch method and liquid-solid extraction with hexane/isopropanol or with hexane/acetone were investigated, as was the effect of previous hydration of samples. Soxhlet extractions were also evaluated with different solvent mixtures. Results on FA composition using the hexane/isopropanol extraction method were the same in terms of FA composition of the Folch method, but the extraction yield was only around 20-40% of that of the Folch method preceded by hydration. Some types of legumes showed particularly interesting values for the ratio of polyunsaturated fatty acids (PUFAs) n-6/n-3, such as lentils, with the value of 4.0, and Azuki beans, at 3.2. In lentils, the PUFAs% ranged from 42.0% to 57.4%, while in Azuki beans it was 57.5%.

  12. Extractive fermentation for enhanced propionic acid production from lactose by Propionibacterium acidipropionici

    SciTech Connect

    Jin, Z.; Yang, S.T.

    1998-05-01

    An extractive fermentation process using an amine extractant and a hollow-fiber membrane extractor to selectively remove propionic acid from the fermentation broth was developed to produce propionate from lactose. Compared to the conventional batch fermentation, the extractive fermentation had a much higher productivity ({approximately}1 g/(L{center_dot}h) or 5-fold increase), higher propionate yield (up to 0.66 g/g or more than 20% increase), higher final product concentration (75 g/L or higher), and higher product purity ({approximately}90%). Meanwhile, acetate and succinate productions in the extractive fermentation were significantly reduced. The improved fermentation performance can be attributed to the reduced product inhibition and a possible metabolic pathway shift to favor more propionic but less acetic and succinic acid production. The process was stable and gave consistent long-term performance over the 1.5-month period studied. The effects of propionate concentration, pH, and amine content in the solvent on the extractive fermentation were also studied and are discussed in this paper.

  13. Integrated printed circuit board device for cell lysis and nucleic acid extraction.

    PubMed

    Marshall, Lewis A; Wu, Liang Li; Babikian, Sarkis; Bachman, Mark; Santiago, Juan G

    2012-11-06

    Preparation of raw, untreated biological samples remains a major challenge in microfluidics. We present a novel microfluidic device based on the integration of printed circuit boards and an isotachophoresis assay for sample preparation of nucleic acids from biological samples. The device has integrated resistive heaters and temperature sensors as well as a 70 μm × 300 μm × 3.7 cm microfluidic channel connecting two 15 μL reservoirs. We demonstrated this device by extracting pathogenic nucleic acids from 1 μL dispensed volume of whole blood spiked with Plasmodium falciparum. We dispensed whole blood directly onto an on-chip reservoir, and the system's integrated heaters simultaneously lysed and mixed the sample. We used isotachophoresis to extract the nucleic acids into a secondary buffer via isotachophoresis. We analyzed the convective mixing action with micro particle image velocimetry (micro-PIV) and verified the purity and amount of extracted nucleic acids using off-chip quantitative polymerase chain reaction (PCR). We achieved a clinically relevant limit of detection of 500 parasites per microliter. The system has no moving parts, and the process is potentially compatible with a wide range of on-chip hybridization or amplification assays.

  14. Nigella sativa L. Seed Extract Modulates the Neurotransmitter Amino Acids Release in Cultured Neurons In Vitro

    PubMed Central

    El-Naggar, Tarek; Gómez-Serranillos, María Pilar; Palomino, Olga María; Arce, Carmen; Carretero, María Emilia

    2010-01-01

    Nigella sativa L. (NS) has been used for medicinal purposes since ancient times. This study aimed to investigate the cytotoxicity of NS dry methanolic extract on cultured cortical neurons and its influence on neurotransmitter release, as well as the presence of excitatory (glutamate and aspartate) and inhibitory amino acids (gamma-aminobutyric acid—GABA—and glycine) in NS extract. Cultured rat cortical neurons were exposed to different times and concentrations of NS dry methanolic extract and cell viability was then determined by a quantitative colorimetric method. NS did not induce any toxicity. The secretion of different amino acids was studied in primary cultured cortical neurons by high-performance liquid chromatography (HPLC) using a derivation before injection with dansyl chloride. NS modulated amino acid release in cultured neurons; GABA was significantly increased whereas secretion of glutamate, aspartate, and glycine were decreased. The in vitro findings support the hypothesis that the sedative and depressive effects of NS observed in vivo could be based on changes of inhibitory/excitatory amino acids levels. PMID:20625485

  15. Carboxylic and dicarboxylic acids extracted from crushed magnesium oxide single crystals

    NASA Technical Reports Server (NTRS)

    Freund, F.; Gupta, A. D.; Kumar, D.

    1999-01-01

    Carboxylic and dicarboxylic acids (glycolic, oxalic, malonic and succinic) have been extracted with tetrahydrofuran (THF) and H2O from large synthetic MgO crystals, crushed to a medium fine powder. The extracts were characterized by infrared spectroscopy and 1H-NMR. The THF extracts were derivatized with tert-butyldimethylsilyl (t-BDMS) for GC-MS analysis. A single crystal separated from the extract was used for an x-ray structure analysis, giving the monoclinic unit cell, space group P21/c with ao = 5.543 A, bo = 8.845 A, co = 5.086 A, and beta = 91.9 degrees, consistent with beta-succinic acid, HOOC(CH2)COOH. The amount of extracted acids is estimated to be of the order of 0.1 to 0.5 mg g-1 MgO. The MgO crystals from which these organic acids were extracted grew from the 2860 degrees C hot melt, saturated with CO/CO2 and H2O, thereby incorporating small amounts of the gaseous components to form a solid solution (ss) with MgO. Upon cooling, the ss becomes supersaturated, causing solute carbon and other solute species to segregate not only to the surface but also internally, to dislocations and subgrain boundaries. The organic acids extracted from the MgO crystals after crushing appear to derive from these segregated solutes that formed C-C, C-H and C-O bonds along dislocations and other defects in the MgO structure, leading to entities that can generically be described as (HxCyOz)n-. The processes underlying the formation of these precursors are fundamental in nature and expected to be operational in any minerals, preferentially those with dense structures, that crystallized in H2O-CO2-laden environments. This opens the possibility that common magmatic and metamorphic rocks when weathering at the surface of a tectonically active planet like Earth may be an important source of abiogenically formed complex organic compounds.

  16. Carboxylic and Dicarboxylic Acids Extracted from Crushed Magnesium Oxide Single Crystals

    NASA Technical Reports Server (NTRS)

    Freund, Friedemann; Gupta, Alka D.; Kumar, Devendra; DeVincenzi, Donald (Technical Monitor)

    1998-01-01

    Carboxylic and dicarboxylic acids (glycolic, oxalic, malonic and succinic) have been extracted with tetrahydrofuran (THE) and H2O from large synthetic MgO crystals, crushed to a medium fine powder. The extracts were characterized by infrared spectroscopy and (sup 1)H-NMR (Nuclear Magnetic Resonance). The THF extracts were derivatized with tert-butyldimethylsilyl (t-BDMS) for GC-MS (Gas Chromatography - Mass Spectroscopy) analysis. A single crystal separated from the extract was used for an x-ray structure analysis, giving the monoclinic unit cell, space group P2(sub 1)/c with a(sub o) = 5.543 A, b(sub o) = 8.845 A, c(sub o) = 5.086 A, and beta = 91.9 degrees, consistent with beta-succinic acid, HOOC(CH2)COOH. The amount of extracted acids is estimated to be of the order of 0.1 to 0.5 mg/g MgO. The MgO crystals from which these organic acids were extracted grew from the 2360 C hot melt, saturated with CO/CO2 and H2O, thereby incorporating small amounts of the gaseous components to form a solid solution (ss) with MgO. Upon cooling, the ss becomes supersaturated, causing solute carbon and other solute species to segregate not only to the surface but also internally, to dislocations and subgrain boundaries. The organic acids extracted from the MgO crystals after crushing appear to derive from these segregated solutes that formed C-C, C-H, and C-O bonds along dislocations and other defects in the MgO structure, leading to entities that can generically be described as (HxCyOz)(sup n-). The processes underlying the formation of these precursors are fundamental in nature and expected to be operational in any minerals, preferentially those with dense structures, that crystallized in H2O-CO2-laden environments. This opens the possibility that common magmatic and metamorphic rocks when weathering at the surface of a tectonically active planet like Earth may be an important source of abiogenically formed complex organic compounds.

  17. Comparison of different commercial DNA extraction kits and PCR protocols for the detection of Echinococcus multilocularis eggs in faecal samples from foxes.

    PubMed

    Maksimov, Pavlo; Schares, Gereon; Press, Sebastian; Fröhlich, Andreas; Basso, Walter; Herzig, Mandy; Conraths, Franz J

    2017-04-15

    Effective and sensitive methods for the molecular detection of Echinococcus multilocularis in faecal samples of final hosts are crucial for the prevention and control of human alveolar echinococcosis and for studies on the epidemiology of the parasite. Little is known about the suitability of commercial test kits for isolating DNA of E. multilocularis from fox faeces and the performance of standard Polymerase Chain Reaction (PCR) protocols in relation to the quality of DNA extracted by these kits. We compared four different kits: ZR Faecal DNA MiniPrep™ (Zymo Research), FastDNA(®) SPIN Kit for Soil (MP Biomedicals), QIAamp(®) Fast DNA Stool Mini Kit (QIAGEN) and NucleoSpin(®) Soil Kit (Macherey-Nagel) for the extraction of DNA from E. multilocularis eggs present in faeces of foxes. Negative faecal samples were spiked with 600, 300, 150, 75, 37, 18, 9, 5 or 2 E. multilocularis eggs, and each egg concentration was tested 10 times with each of the DNA extraction kits. Each extracted DNA sample was amplified using three PCR protocols: i. conventional PCR (cPCR, Platinum(®)Taq, Invitrogen), ii. qPCR with the iQ™ Supermix (Bio-Rad) and iii. qPCR with the QuantiTect(®) Multiplex-Master Mix (QIAGEN). The highest analytical sensitivities for molecular detection of E. multilocularis eggs in spiked fox faeces were observed when combining either the QIAamp(®) Fast DNA Stool Mini Kit or the ZR Faecal DNA MiniPrep™ kit with the qPCR using the QuantiTect(®) Multiplex-Master Mix (Sensitivities 97% and 94%, respectively). Combinations including the remaining test kits (NucleoSpin(®) Soil Kit and FastDNA(®) SPIN Kit for Soil) showed a markedly lower analytical sensitivity for PCR examinations. The results of the present study indicate that it is of utmost importance to select suitable DNA extraction kits in combination with robust PCR methods or reagents to achieve acceptable analytical sensitivity in the molecular detection of E. multilocularis eggs in fox faecal

  18. Extraction and quantification of gymnemic acids through gymnemagenin from callus cultures of Gymnema sylvestre.

    PubMed

    Kanetkar, P V; Singhal, R S; Laddha, K S; Kamat, M Y

    2006-01-01

    The phyto-constituents of Gymnema sylvestre are used in the treatment of diabetes and obesity. The present work reports on the extraction of gymnemic acid through gymnemagenin from callus cultures of G. sylvestre. Components were separated on pre-coated silica gel 60 GF254 plates with chloroform:methanol (8:2) and scanned using a densitometric scanner at 205 nm in the near-UV region. Linearity of determination of gymnemagenin was observed in the range 2-10 microg. The average percentage recovery of gymnemagenin from leaf callus extracts was 98.9+/-0.3.

  19. Chlorogenic acids from green coffee extract are highly bioavailable in humans.

    PubMed

    Farah, Adriana; Monteiro, Mariana; Donangelo, Carmen M; Lafay, Sophie

    2008-12-01

    Chlorogenic acids (CGA) are cinnamic acid derivatives with biological effects mostly related to their antioxidant and antiinflammatory activities. Caffeoylquinic acids (CQA) and dicaffeoylquinic acids (diCQA) are the main CGA found in nature. Because green coffee is a major source of CGA, it has been used for production of nutraceuticals. However, data on the bioavailability of CGA from green coffee in humans are inexistent. The present study evaluated the pharmacokinetic profile and apparent bioavailability of CGA in plasma and urine of 10 healthy adults for 8 h after the consumption of a decaffeinated green coffee extract containing 170 mg of CGA. Three CQA, 3 diCQA, and caffeic, ferulic, isoferulic, and p-coumaric acids were identified in plasma by HPLC-Diode Array Detector-MS after treatment. Over 30% (33.1 +/- 23.1%) of the ingested cinnamic acid moieties were recovered in plasma, including metabolites, with peak levels from 0.5 to 8 h after treatment. CGA and metabolites identified in urine after treatment were 4-CQA, 5-CQA, and sinapic, p-hydroxybenzoic, gallic, vanillic, dihydrocaffeic, caffeic, ferulic, isoferulic, and p-coumaric acids, totaling 5.5 +/- 10.6% urinary recovery of the ingested cinnamic and quinic acid moiteties. This study shows that the major CGA compounds present in green coffee are highly absorbed and metabolized in humans.

  20. Applicability of a System for fully automated nucleic acid extraction from formalin-fixed paraffin-embedded sections for routine KRAS mutation testing.

    PubMed

    Lehmann, Annika; Schewe, Christiane; Hennig, Guido; Denkert, Carsten; Weichert, Wilko; Budczies, Jan; Dietel, Manfred

    2012-06-01

    Due to the approval of various new targeted therapies for the treatment of cancer, molecular pathology laboratories with a diagnostic focus have to meet new challenges: simultaneous handling of a large number of samples, small amounts of input material, and fragmentation of nucleic acids because of formalin fixation. As a consequence, fully automated systems for a fast and standardized extraction of high-quality DNA from formalin-fixed paraffin-embedded (FFPE) tissues are urgently needed. In this study, we tested the performance of a fully automated, high-throughput method for the extraction of nucleic acids from FFPE tissues. We investigated the extraction performance in sections of 5 different tissue types often analyzed in routine pathology laboratories (cervix, colon, liver, lymph node, and lung; n=340). Furthermore, we compared the quality, labor input, and applicability of the method for diagnostic purposes with those of a laboratory-validated manual method in a clinical setting by screening a set of 45 colorectal adenocarcinoma for the KRAS mutation. Automated extraction of both DNA and RNA was successful in 339 of 340 FFPE samples representing 5 different tissue types. In comparison with a conventional manual extraction protocol, the method showed an overall agreement of 97.7% (95% confidence interval, 88.2%-99.9%) for the subsequent mutational analysis of the KRAS gene in colorectal cancer samples. The fully automated system is a promising tool for a simple, robust, and rapid extraction of DNA and RNA from formalin-fixed tissue. It ensures a standardization of sample processing and can be applied to clinical FFPE samples in routine pathology.

  1. Prospective evaluation of a new automated nucleic acid extraction system using routine clinical respiratory specimens.

    PubMed

    Mengelle, C; Mansuy, J-M; Sandres-Sauné, K; Barthe, C; Boineau, J; Izopet, J

    2012-06-01

    The aim of the study was to evaluate the MagNA Pure 96™ nucleic acid extraction system using clinical respiratory specimens for identifying viruses by qualitative real-time PCR assays. Three extraction methods were tested, that is, the MagNA Pure LC™, the COBAS Ampliprep™, and the MagNA Pure 96™ with 10-fold dilutions of an influenza A(H1N1)pdm09 sample. Two hundred thirty-nine respiratory specimens, 35 throat swabs, 164 nasopharyngeal specimens, and 40 broncho-alveolar fluids, were extracted with the MagNA Pure 96™ and the COBAS Ampliprep™ instruments. Forty COBAS Ampliprep™ positive samples were also tested. Real-time PCRs were used to identify influenza A and influenza A(H1N1)pdm09, rhinovirus, enterovirus, adenovirus, varicella zoster virus, cytomegalovirus, and herpes simplex virus. Similar results were obtained on RNA extracted from dilutions of influenza A(H1N1)pdm09 with the three systems: the MagNA Pure LC™, the COBAS Ampliprep™, and the MagNA Pure 96™. Data from clinical respiratory specimens extracted with the MagNA Pure 96™ and COBAS Ampliprep™ instruments were in 98.5% in agreement (P < 0.0001) for influenza A and influenza A(H1N1)pdm09. Data for rhinovirus were in 97.3% agreement (P < 0.0001) and in 96.8% agreement for enterovirus. They were in 100% agreement for adenovirus. Data for cytomegalovirus and HSV1-2 were in 95.2% agreement (P < 0.0001). The MagNA Pure 96™ instrument is easy-to-use, reliable, and has a high throughput for extracting total nucleic acid from respiratory specimens. These extracts are suitable for molecular diagnosis with any type of real-time PCR assay.

  2. New optimized DNA extraction protocol for fingerprints deposited on a special self-adhesive security seal and other latent samples used for human identification.

    PubMed

    Kopka, Julieta; Leder, Monika; Jaureguiberry, Stella M; Brem, Gottfried; Boselli, Gabriel O

    2011-09-01

    Obtaining complete short tandem repeat (STR) profiles from fingerprints containing minimal amounts of DNA, using standard extraction techniques, can be difficult. The aim of this study was to evaluate a new kit, Fingerprint DNA Finder (FDF Kit), recently launched for the extraction of DNA and STR profiling from fingerprints placed on a special device known as Self-Adhesive Security Seal Sticker(®) and other latent fingerprints on forensic evidentiary material like metallic guns. The DNA extraction system is based on a reversal of the silica principle, and all the potential inhibiting substances are retained on the surface of a special adsorbent, while nucleic acids are not bound and remain in solution dramatically improving DNA recovery. DNA yield was quite variable among the samples tested, rendering in most of the cases (>90%) complete STR profiles, free of PCR inhibitors, and devoid of artifacts. Even samples with DNA amount below 100 pg could be successfully analyzed.

  3. Biochemical characteristics and gelling capacity of pectin from yellow passion fruit rind as affected by acid extractant nature.

    PubMed

    Yapo, Beda M

    2009-02-25

    The effects of acid extractant type on the yield and characteristics of pectin from yellow passion fruit (Passiflora edulis flavicarpa) rind was investigated by using citric, nitric, or sulfuric acids at different concentrations (10 mM and 30 mM) and pH (1.8 and 2.5). The results showed that not only concentration, but also acid type influenced the extracted pectin yields (3-14%, w/w). The yield of pectin extracted with citric acid was the lowest. Acid type and concentration affected the molecular characteristics of pectin, notably, the degree of esterification (29-73), galacturonic acid to rhamnose ratio (14-35), weight average-molecular weight (100-250 kDa), gel strength (127-179), and setting time (841-1236 s). Citric acid-extracted pectin had a higher degree of esterification and weight average-molecular weight and better gelling properties. At 30 mM concentration, nitric and sulfuric acids solubilize pectins having a degree of esterification <50, contrary to citric acid. The results indicate that the latter acid exerts the least deesterifying action on pectin solubilization from the cell wall material. Citric acid-extracted pectin was closer to lemon pectin of similar degree of esterification in terms of gelling properties.

  4. A fully disposable and integrated paper-based device for nucleic acid extraction, amplification and detection.

    PubMed

    Tang, Ruihua; Yang, Hui; Gong, Yan; You, MinLi; Liu, Zhi; Choi, Jane Ru; Wen, Ting; Qu, Zhiguo; Mei, Qibing; Xu, Feng

    2017-03-29

    Nucleic acid testing (NAT) has been widely used for disease diagnosis, food safety control and environmental monitoring. At present, NAT mainly involves nucleic acid extraction, amplification and detection steps that heavily rely on large equipment and skilled workers, making the test expensive, time-consuming, and thus less suitable for point-of-care (POC) applications. With advances in paper-based microfluidic technologies, various integrated paper-based devices have recently been developed for NAT, which however require off-chip reagent storage, complex operation steps and equipment-dependent nucleic acid amplification, restricting their use for POC testing. To overcome these challenges, we demonstrate a fully disposable and integrated paper-based sample-in-answer-out device for NAT by integrating nucleic acid extraction, helicase-dependent isothermal amplification and lateral flow assay detection into one paper device. This simple device allows on-chip dried reagent storage and equipment-free nucleic acid amplification with simple operation steps, which could be performed by untrained users in remote settings. The proposed device consists of a sponge-based reservoir and a paper-based valve for nucleic acid extraction, an integrated battery, a PTC ultrathin heater, temperature control switch and on-chip dried enzyme mix storage for isothermal amplification, and a lateral flow test strip for naked-eye detection. It can sensitively detect Salmonella typhimurium, as a model target, with a detection limit of as low as 10(2) CFU ml(-1) in wastewater and egg, and 10(3) CFU ml(-1) in milk and juice in about an hour. This fully disposable and integrated paper-based device has great potential for future POC applications in resource-limited settings.

  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.

  6. Biotransformation of caffeoyl quinic acids from green coffee extracts by Lactobacillus johnsonii NCC 533

    PubMed Central

    2013-01-01

    The potential of Lactobacillus johnsonii NCC 533 to metabolize chlorogenic acids from green coffee extract was investigated. Two enzymes, an esterase and a hydroxycinnamate decarboxylase (HCD), were involved in this biotransformation. The complete hydrolysis of 5-caffeoylquinic acid (5-CQA) into caffeic acid (CA) by L. johnsonii esterase occurred during the first 16 h of reaction time. No dihydrocaffeic acid was identified in the reaction mixture. The decarboxylation of CA into 4-vinylcatechol (4-VC) started only when the maximum concentration of CA was reached (10 μmol/ml). CA was completely transformed into 4-VC after 48 h of incubation. No 4-vinylphenol or other derivatives could be identified in the reaction media. In this study we demonstrate the capability of L. johnsonii to transform chlorogenic acids from green coffee extract into 4-VC in two steps one pot reaction. Thus, the enzymatic potential of certain lactobacilli might be explored to generate flavor compounds from plant polyphenols. PMID:23692950

  7. Kinetics of the extraction of succinic acid with tri-n-octylamine in 1-octanol solution.

    PubMed

    Jun, Young-Si; Huh, Yun Suk; Hong, Won Hi; Hong, Yeon Ki

    2005-01-01

    Kinetic studies for the extraction of succinic acid from aqueous solution with 1-octanol solutions of tri-n-octylamine (TOA) were carried out using a stirred cell with a microporous hydrophobic membrane. The interfacial concentrations of species were correlated and thus the intrinsic kinetics was obtained. The overall extraction process was controlled by the chemical reaction at or near the interface between the aqueous and organic phases. The formation reaction of succinic acid-TOA complex was found to be first order with respect to the concentration of succinic acid in the aqueous phase and the order of 0.5 with respect to that of TOA in the organic phase with a rate constant of (3.14 +/- 0.6) x 10(-8) m(2.5) x mol(-0.5) x s(-1). The dissociation reaction of succinic acid-TOA complex was found to be the second-order with respect to that of succinic acid-TOA complex in the organic phase and the order of -2 with respect to that of TOA in the organic phase with a rate constant of (1.44 +/- 1.4) x 10(-4) mol x m(-2) x s(-1).

  8. Extraction of Alumina from high-silica bauxite by hydrochloric acid leaching using preliminary roasting method

    NASA Astrophysics Data System (ADS)

    Valeev, D. V.; Mansurova, E. R.; Bychinskii, V. A.; Chudnenko, K. V.

    2016-02-01

    A process of dissolution Severoonezhsk deposit boehmite-kaolinite bauxite by hydrochloric acid, as well as the processes that occur during open-air calcination, were investigated. A dehydration process has been studied, and the basic phase transformation temperatures were identified. Temperature and time of calcination influence on bauxite dehydration speed were determined. It is shown that the preliminary calcination increases the extraction ratio of alumina into solution up to 89%. Thermodynamic modelling of physical and chemical processes of bauxite decomposition by hydrochloric acid and the basic forms of aluminium speciation in solution were obtained.

  9. Simplified protocol for DNA extraction and amplification of 2 molecular markers to detect and type Giardia duodenalis.

    PubMed

    Uda-Shimoda, Carla Fernanda; Colli, Cristiane Maria; Pavanelli, Mariana Felgueira; Falavigna-Guilherme, Ana Lúcia; Gomes, Mônica Lúcia

    2014-01-01

    We evaluated the ability of 3 kits: QIAmp® DNA stool mini kit (Qiagen, Hilden, Germany), PureLink PCR Purification®, and PureLink™ Genomic DNA® (Invitrogen, Carlsbad, CA, USA) for DNA extraction, and of 2 molecular markers (heat shock protein [HSP] and β-giardin genes) for detection and genotyping of Giardia duodenalis stool samples. The detection and typing limits of the markers were determined by the DNA concentration of trophozoites and cysts and were tested in 26 clinical samples. Of the 3 kits tested, the PureLink PCR Purification gave the best results when tested with clinical samples with low, intermediate, and high numbers of cysts. The DNA extracted from trophozoites and cysts was diluted successively in 1:2 ratios until it was no longer possible to observe the amplified product in polyacrylamide gel. Similarly, a suspension of cysts was diluted until no cysts were observed, and then the DNA was extracted. The amount of DNA of trophozoites and cysts for the typing of the parasite was smaller for the HSP marker than for β-giardin. Combined use of both markers allowed us to detect DNA of Giardia in parasitologically positive samples in a higher percentage (75%) than the results obtained for each marker and in 1 parasitologically negative sample, indicating that this combination increased the potential to accurately detect and genotype this parasite. We also concluded that the HSP marker has a higher limit of detection and typing than the β-giardin marker and that the DNA extraction method tested for G. duodenalis is simpler and more efficient than those that are currently in use and can be applied on a large scale.

  10. Comparison of manual and semi-automatic DNA extraction protocols for the barcoding characterization of hematophagous louse flies (Diptera: Hippoboscidae).

    PubMed

    Gutiérrez-López, Rafael; Martínez-de la Puente, Josué; Gangoso, Laura; Soriguer, Ramón C; Figuerola, Jordi

    2015-06-01

    The barcoding of life initiative provides a universal molecular tool to distinguish animal species based on the amplification and sequencing of a fragment of the subunit 1 of the cytochrome oxidase (COI) gene. Obtaining good quality DNA for barcoding purposes is a limiting factor, especially in studies conducted on small-sized samples or those requiring the maintenance of the organism as a voucher. In this study, we compared the number of positive amplifications and the quality of the sequences obtained using DNA extraction methods that also differ in their economic costs and time requirements and we applied them for the genetic characterization of louse flies. Four DNA extraction methods were studied: chloroform/isoamyl alcohol, HotShot procedure, Qiagen DNeasy(®) Tissue and Blood Kit and DNA Kit Maxwell(®) 16LEV. All the louse flies were morphologically identified as Ornithophila gestroi and a single COI-based haplotype was identified. The number of positive amplifications did not differ significantly among DNA extraction procedures. However, the quality of the sequences was significantly lower for the case of the chloroform/isoamyl alcohol procedure with respect to the rest of methods tested here. These results may be useful for the genetic characterization of louse flies, leaving most of the remaining insect as a voucher.

  11. Selective precipitation of ribonucleic acid from a mixture of total cellular nucleic acids extracted from cultured mammalian cells

    PubMed Central

    Harrison, P. R.

    1971-01-01

    A simple and reproducible method is described for precipitating RNA selectively from total mammalian-cell nucleic acids extracted by the phenol–sodium dodecyl sulphate procedure at pH8.0. Under specified conditions bulk RNA is precipitated almost quantitatively whereas bulk DNA remains in solution. Minor components of RNA (detected by pulse-labelling and chromatography on methylated albumin–kieselguhr) and rapidly labelled components of DNA containing single-stranded regions are also precipitated. The usefulness of the method is discussed in the context of isolating separately both RNA and DNA from cultured cells that are difficult to obtain in quantity. PMID:5165620

  12. Spontaneous surface convection in extraction of lanthanoids by di-2-ethylhexylphosphoric acid

    SciTech Connect

    Dupal, A.Ya.; Tarasov, V.V.; Yagodin, G.A.; Arutyunyan, V.A.

    1988-09-01

    It has been established that when lanthanoids are extracted from aqueous nitric acid solutions (pH > 1.5) by di-2-ethylhexylphosphoric acid in decane or toluene a spontaneous surface convection occurs in the system over the initial period, which leads to an up to tenfold increase in the mass transfer coefficient. The intensity of the spontaneous surface convection (SSC) depends on the concentrations of the components and the conditions under which the extraction is conducted. With the passage of time an interphase film is formed at the interface which suppresses the SSC and retards the mass transfer. Small additions of ionic surfactants increase the surface viscosity, reducing any motion in the interphase region, which leads to an effective suppression of the SSC.

  13. Recovery of Uranium from Wet Phosphoric Acid by Solvent Extraction Processes

    SciTech Connect

    Beltrami, Denis; Cote, Gérard; Mokhtari, Hamid; Courtaud, Bruno; Moyer, Bruce A.; Chagnes, Alexandre

    2014-11-17

    Between 1951 and 1991, we developed about 17 processes to recover uranium from wet phosphoric acid (WPA), but the viability of these processes was subject to the variation of the uranium price market. Nowadays, uranium from WPA appears to be attractive due to the increase of the global uranium demand resulting from the emergence of developing countries. Moreover, the increasing demand provides impetus for a new look at the applicable technology with a view to improvements as well as altogether new approaches. This paper gives an overview on extraction processes developed in the past to recover uranium from wet phosphoric acid (WPA) as well as the physicochemistry involved in these processes. Recent advances concerning the development of new extraction systems are also reported and discussed.

  14. Recovery of Uranium from Wet Phosphoric Acid by Solvent Extraction Processes

    DOE PAGES

    Beltrami, Denis; Cote, Gérard; Mokhtari, Hamid; ...

    2014-11-17

    Between 1951 and 1991, we developed about 17 processes to recover uranium from wet phosphoric acid (WPA), but the viability of these processes was subject to the variation of the uranium price market. Nowadays, uranium from WPA appears to be attractive due to the increase of the global uranium demand resulting from the emergence of developing countries. Moreover, the increasing demand provides impetus for a new look at the applicable technology with a view to improvements as well as altogether new approaches. This paper gives an overview on extraction processes developed in the past to recover uranium from wet phosphoricmore » acid (WPA) as well as the physicochemistry involved in these processes. Recent advances concerning the development of new extraction systems are also reported and discussed.« less

  15. Supercritical carbon dioxide extraction of seed oil from winter melon (Benincasa hispida) and its antioxidant activity and fatty acid composition.

    PubMed

    Bimakr, Mandana; Rahman, Russly Abdul; Taip, Farah Saleena; Adzahan, Noranizan Mohd; Sarker, Md Zaidul Islam; Ganjloo, Ali

    2013-01-15

    In the present study, supercritical carbon dioxide (SC-CO(2)) extraction of seed oil from winter melon (Benincasa hispida) was investigated. The effects of process variables namely pressure (150-300 bar), temperature (40-50 °C) and dynamic extraction time (60-120 min) on crude extraction yield (CEY) were studied through response surface methodology (RSM). The SC-CO(2) extraction process was modified using ethanol (99.9%) as co-solvent. Perturbation plot revealed the significant effect of all process variables on the CEY. A central composite design (CCD) was used to optimize the process conditions to achieve maximum CEY. The optimum conditions were 244 bar pressure, 46 °C temperature and 97 min dynamic extraction time. Under these optimal conditions, the CEY was predicted to be 176.30 mg-extract/g-dried sample. The validation experiment results agreed with the predicted value. The antioxidant activity and fatty acid composition of crude oil obtained under optimized conditions were determined and compared with published results using Soxhlet extraction (SE) and ultrasound assisted extraction (UAE). It was found that the antioxidant activity of the extract obtained by SC-CO(2) extraction was strongly higher than those obtained by SE and UAE. Identification of fatty acid composition using gas chromatography (GC) showed that all the extracts were rich in unsaturated fatty acids with the most being linoleic acid. In contrast, the amount of saturated fatty acids extracted by SE was higher than that extracted under optimized SC-CO(2) extraction conditions.

  16. Interlaboratory comparison of measurements of acid-volatile sulfide and simultaneously extracted nickel in spiked sediments

    USGS Publications Warehouse

    Brumbaugh, William G.; Hammerschmidt, Chad R.; Zanella, Luciana; Rogevich, Emily; Salata, Gregory; Bolek, Radoslaw

    2011-01-01

    An interlaboratory comparison of acid-volatile sulfide (AVS) and simultaneously extracted nickel (SEM_Ni) measurements of sediments was conducted among five independent laboratories. Relative standard deviations for the seven test samples ranged from 5.6 to 71% (mean = 25%) for AVS and from 5.5 to 15% (mean = 10%) for SEM_Ni. These results are in stark contrast to a recently published study that indicated AVS and SEM analyses were highly variable among laboratories.

  17. [Determination of phthalic acid esters in textiles by solid phase extraction-gas chromatography].

    PubMed

    Niu, Zengyuan; Ye, Xiwen; Fang, Liping; Xue, Qiuhong; Sun, Zhongsong

    2006-09-01

    A method was established for the simultaneous determination of some phthalic acid esters, namely, dimethyl phthalate (DMP), diethyl phthalate (DEP), dipropyl phthalate (DPrP), dibutyl phthalate (DBP), diamyl phthalate (DAP), dihexyl phthalate (DHP), benzyln-butyl phthalate (BBP), di-(2-ethylhexyl) phthalate (DEHP), dicyclohexyl phthalate (DCHP), di-n-octyl phthalate (DNOP), diisononyl phthalate (DINP) and diisodecyl phthalate (DIDP) in textiles by solid phase extraction (SPE) coupled with gas chromatography (GC). The phthalic acid esters in textiles were extracted by Soxhlet extraction with hexane, the extracts were then cleaned up and enriched by a strong anion exchange (SAX) SPE cartridge. The parameters affecting the purification efficiency of SPE cartridge, such as solvent conditioning, rinsing, and elution, were studied. Conditioning with 5 mL hexane and rinsing with 3 mL isooctane were proved to be the optimal conditions. Of the several solvent ratios (ethylacetate in hexane) used for selective elution of phthalic acid esters from the SAX SPE cartridge, the 15% (v/v) content for ethylacetate in hexane gave the best result. Under the optimized conditions, the recoveries of phthalic acid esters for spiked standards (n=7) were 86.3%-102.7%, and the relative standard deviations (RSDs) were less than 5%. In this method the detection limits for DMP, DEP, DPrP, DBP, DAP, BBP, DCHP, DEHP, DNOP were all below 1 mg/kg, and the detection limits for DINP and DIDP were 1.74 mg/kg and 1.55 mg/kg respectively. This SPE-GC method is sensitive, accurate and suitable for the analysis of phthalate environmental hormones in textiles.

  18. Solvent extraction of organic acids from stillage for its re-use in ethanol production process.

    PubMed

    Castro, G A; Caicedo, L A; Alméciga-Díaz, C J; Sanchez, O F

    2010-06-01

    Stillage re-use in the fermentation stage in ethanol production is a technique used for the reduction of water and fermentation nutrients consumption. However, the inhibitory effect on yeast growth of the by-products and feed components that remains in stillage increases with re-use and reduces the number of possible recycles. Several methods such as ultrafiltration, electrodialysis and advanced oxidation processes have been used in stillage treatment prior its re-use in the fermentation stage. Nevertheless, few studies evaluating the effect of solvent extraction as a stillage treatment option have been performed. In this work, the inhibitory effect of serial stillage recycling over ethanol and biomass production was determined, using acetic acid as a monitoring compound during the fermentation and solvent extraction process. Raw palm oil methyl ester showed the highest acetic acid extraction from the aqueous phase, presenting a distribution coefficient of 3.10 for a 1:1 aqueous phase mixture:solvent ratio. Re-using stillage without treatment allowed up to three recycles with an ethanol production of 53.7 +/- 2.0 g L(-1), which was reduced 25% in the fifth recycle. Alternatively, treated stillage allowed up to five recycles with an ethanol final concentration of 54.7 +/- 1.3 g L(- 1). These results show that reduction of acetic acid concentration by an extraction process with raw palm oil methyl ester before re-using stillage improves the number of recycles without a major effect on ethanol production. The proposed process generates a palm oil methyl ester that contains organic acids, among other by-products, that could be used for product recovery and as an alternative fuel.

  19. Gas chromatographic determination of urinary phenol conjugates after acid hydrolysis/extractive acetylation.

    PubMed

    Weber, L

    1992-02-14

    Phenolic metabolites of inhaled aromatic solvent vapours were liberated by acid hydrolysis of their urinary conjugates. Steam distillation enhanced by salting-out with MgSO4 gave good recoveries. After extractive acetylation, the derivatives of all cresols and xylenols were completely separated on a Se-54 capillary column. The overall recoveries of urinary phenols relative to the internal standard, 3-chlorophenol, were in the range 92-99%.

  20. Interlaboratory comparison of measurements of acid-volatile sulfide and simultaneously extracted nickel in spiked sediments

    USGS Publications Warehouse

    Brumbaugh, W.G.; Hammerschmidt, C.R.; Zanella, L.; Rogevich, E.; Salata, G.; Bolek, R.

    2011-01-01

    An interlaboratory comparison of acid-volatile sulfide (AVS) and simultaneously extracted nickel (SEM-Ni) measurements of sediments was conducted among five independent laboratories. Relative standard deviations for the seven test samples ranged from 5.6 to 71% (mean=25%) for AVS and from 5.5 to 15% (mean=10%) for SEM-Ni. These results are in stark contrast to a recently published study that indicated AVS and SEM analyses were highly variable among laboratories. ?? 2011 SETAC.

  1. Interlaboratory comparison of measurements of acid-volatile sulfide and simultaneously extracted nickel in spiked sediments

    USGS Publications Warehouse

    Brumbaugh, William G.; Hammerschmidt, Chad R.; Zanella, Luciana; Rogevich, Emily; Salata, Gregory; Bolek, Radoslaw

    2011-01-01

    An interlaboratory comparison of acid-volatile sulfide (AVS) and simultaneously extracted nickel (SEM_Ni) measurements of sediments was conducted among five independent laboratories. Relative standard deviations for the seven test samples ranged from 5.6 to 71% (mean?=?25%) for AVS and from 5.5 to 15% (mean?=?10%) for SEM_Ni. These results are in stark contrast to a recently published study that indicated AVS and SEM analyses were highly variable among laboratories.

  2. Extraction of metals and/or metalloids from acidic media using supercritical fluids and salts

    DOEpatents

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

    1998-01-01

    A method of extracting metalloid and metal species from a solid or liquid material by exposing the material to a fluid solvent, particularly supercritical carbon dioxide, containing a chelating agent is described. The chelating agent forms chelates that are soluble in the fluid to allow removal of the species from the material. In preferred embodiments, the extraction solvent is supercritical carbon dioxide and the chelating agent comprises a trialkyl phosphate, a triaryl phosphate, a trialkylphosphine oxide, a triarylphosphine oxide, or mixtures thereof. The method provides an environmentally benign process for removing contaminants from industrial waste. The method is particularly useful for extracting actinides from acidic solutions, and the process can be aided by the addition of nitrate salts. The chelate and supercritical fluid can be regenerated, and the contaminant species recovered, to provide an economic, efficient process.

  3. Extraction of metals and/or metalloids from acidic media using supercritical fluids and salts

    DOEpatents

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

    1998-06-23

    A method is described for extracting metalloid and metal species from a solid or liquid material by exposing the material to a fluid solvent, particularly supercritical carbon dioxide, containing a chelating agent. The chelating agent forms chelates that are soluble in the fluid to allow removal of the species from the material. In preferred embodiments, the extraction solvent is supercritical carbon dioxide and the chelating agent comprises a trialkyl phosphate, a triaryl phosphate, a trialkylphosphine oxide, a triarylphosphine oxide, or mixtures thereof. The method provides an environmentally benign process for removing contaminants from industrial waste. The method is particularly useful for extracting actinides from acidic solutions, and the process can be aided by the addition of nitrate salts. The chelate and supercritical fluid can be regenerated, and the contaminant species recovered, to provide an economic, efficient process. 7 figs.

  4. Dyeing wool and cotton fibres with acidic extract of Hibiscus rosa sinensis flower.

    PubMed

    Hayat, Lamya; Jacob, Dangly Ann

    2016-05-02

    The focus of this work is to extract a natural dye for colouring camel wool as a substitute for synthetic dyes used in the Sadu House of Kuwait. Their target is to keep the tradition of tent and rug production natural in all its manifestations. Therefore, our task was to find an abundant source that provides a colour preferably red to purple. Hibiscus rosa sinensis (HRS) is an abundantly available plant in Kuwait that was explored for extraction of the red dye to colour camel wool permanently. The powdered petals of red flowers of HRS was extracted with 5% acetic acid which yielded a deep red colour that showed a great potential for woollen fibre dyeing. The use of mordants like alum and some metal salts manifested a wide range of fixed colours which intensified at 85 °C. The colours produced had excellent fastness and was accepted by the Sadu House.

  5. Hypoglycaemic effects of tea extracts and ent-kaurenoic acid from Smallanthus sonchifolius.

    PubMed

    Raga, Dennis D; Alimboyoguen, Agnes B; del Fierro, Ramon S; Ragasa, Consolacion Y

    2010-11-01

    Hypoglycaemic activity was observed in normoglycaemic mice orally administered with the aqueous Smallanthus sonchifolius leaf tea extract, alloxan-induced diabetic mice orally administered with ent-kaurenoic acid (1), and normoglycaemic mice intraperitoneally administered with 1 from S. sonchifolius leaves. A single dose administration of 50 mg kg(-1) BW yacon leaf tea extract demonstrated immediate but relatively short hypoglycaemic activity, with significant effects observed during 1-2 h. Similarly, administration with 100 mg kg(-1) BW yacon leaf tea extract obtained by heavy stirring in hot water demonstrated a more potent activity compared to the positive control at 1.5-2.0 h. Oral administration of 1 did not affect the blood glucose level of the alloxan-induced diabetic mice, but a single intraperitonial injection of 10 mg kg(-1) BW in normoglycaemic mice had consistent percent blood glucose reduction persisting from 1 to 2 h observation periods.

  6. [Determination of trace haloacetic acids in drinking water using ion chromatography coupled with solid phase extraction].

    PubMed

    Sun, Yingxue; Huang, Jianjun; Gu, Ping

    2006-05-01

    The combined solid phase extraction (SPE)-ion chromatography (IC) method was developed for the analysis of trace haloacetic acids (HAAs) in drinking water. The tested HAAs included monochloroacetic acid (MCAA), dichloroacetic acid (DCAA), trichloroacetic acid (TCAA), monobromoacetic acid (MBAA) and dibromoacetic acid (DBAA). For trace determination of HAAs in real drinking water samples, conditions of LiChrolut EN SPE cartridge were investigated for HAAs preconcentration and matrix elimination. Elution was carried out by 2 mL of sodium hydroxide (10 mmol/L) with the flow rate of 2 mL/min. The Dionex IonPac AS16 column (250 mm x 4 mm i. d.), a high capacity and hydroxide-selective anion-exchange column designed for the determination of polarizable anions, was chosen for chromatographic separation. HAAs were analyzed with a concentration gradient of NaOH with the flow rate of 0.8 mL/min and detected by suppressed conductivity. A 500 microL sample loop was used. The detection limits of this SPE-IC method for MCAA, DCAA, DBAA and TCAA were 0.38-1.69 microg/L and MBAA was 12.5 microg/L under 25-fold preconcentration. The results demonstrate that the method is suitable for the analysis of trace haloacetic acids in drinking water.

  7. Addition of Grape Seed Extract Renders Phosphoric Acid a Collagen-stabilizing Etchant.

    PubMed

    Liu, Y; Dusevich, V; Wang, Y

    2014-08-01

    Previous studies found that grape seed extract (GSE), which is rich in proanthocyanidins, could protect demineralized dentin collagen from collagenolytic activities following clinically relevant treatment. Because of proanthocyanidin's adverse interference to resin polymerization, it was believed that GSE should be applied and then rinsed off in a separate step, which in effect increases the complexity of the bonding procedure. The present study aimed to investigate the feasibility of combining GSE treatment with phosphoric acid etching to address the issue. It is also the first attempt to formulate collagen-cross-linking dental etchants. Based on Fourier-transformed infrared spectroscopy and digestion assay, it was established that in the presence of 20% to 5% phosphoric acid, 30 sec of GSE treatment rendered demineralized dentin collagen inert to bacterial collagenase digestion. Based on this positive result, the simultaneous dentin etching and collagen protecting of GSE-containing phosphoric acid was evaluated on the premise of a 30-second etching time. According to micro-Raman spectroscopy, the formulation containing 20% phosphoric acid was found to lead to overetching. Based on scanning and transmission electronic microscopy, this same formulation exhibited unsynchronized phosphoric acid and GSE penetration. Therefore, addition of GSE did render phosphoric acid a collagen-stabilizing etchant, but the preferable phosphoric acid concentration should be <20%.

  8. Analysis of perfluorinated carboxylic acids in soils II: optimization of chromatography and extraction.

    PubMed

    Washington, John W; Henderson, W Matthew; Ellington, J Jackson; Jenkins, Thomas M; Evans, John J

    2008-02-15

    With the objective of detecting and quantitating low concentrations of perfluorinated carboxylic acids (PFCAs), including perfluorooctanoic acid (PFOA), in soils, we compared the analytical suitability of liquid chromatography columns containing three different stationary phases, two different liquid chromatography-tandem mass spectrometry (LC/MS/MS) systems, and eight combinations of sample-extract pretreatments, extractions and cleanups on three test soils. For the columns and systems we tested, we achieved the greatest analytical sensitivity for PFCAs using a column with a C(18) stationary phase in a Waters LC/MS/MS. In this system we achieved an instrument detection limit for PFOA of 270 ag/microL, equating to about 14 fg of PFOA on-column. While an elementary acetonitrile/water extraction of soils recovers PFCAs effectively, natural soil organic matter also dissolved in the extracts commonly imparts significant noise that appears as broad, multi-nodal, asymmetric peaks that coelute with several PFCAs. The intensity and elution profile of this noise is highly variable among soils and it challenges detection of low concentrations of PFCAs by decreasing the signal-to-noise contrast. In an effort to decrease this background noise, we investigated several methods of pretreatment, extraction and cleanup, in a variety of combinations, that used alkaline and unbuffered water, acetonitrile, tetrabutylammonium hydrogen sulfate, methyl-tert-butyl ether, dispersed activated carbon and solid-phase extraction. For the combined objectives of complete recovery and minimization of background noise, we have chosen: (1) alkaline pretreatment; (2) extraction with acetonitrile/water; (3) evaporation to dryness; (4) reconstitution with tetrabutylammonium-hydrogen-sulfate ion-pairing solution; (5) ion-pair extraction to methyl-tert-butyl ether; (6) evaporation to dryness; (7) reconstitution with 60/40 acetonitrile/water (v/v); and (8) analysis by LC/MS/MS. Using this method, we

  9. Selective extraction of zinc(II) over iron(II) from spent hydrochloric acid pickling effluents by liquid-liquid extraction.

    PubMed

    Mansur, Marcelo Borges; Rocha, Sônia Denise Ferreira; Magalhães, Fernando Silva; Benedetto, Jeaneth dos Santos

    2008-02-11

    The selective removal of zinc(II) over iron(II) by liquid-liquid extraction from spent hydrochloric acid pickling effluents produced by the zinc hot-dip galvanizing industry was studied at room temperature. Two distinct effluents were investigated: effluent 1 containing 70.2g/L of Zn, 92.2g/L of Fe and pH 0.6, and effluent 2 containing 33.9 g/L of Zn, 203.9g/L of Fe and 2M HCl. The following extractants were compared: TBP (tri-n-butyl phosphate), Cyanex 272 [bis(2,4,4-trimethylpentyl)phosphinic acid], Cyanex 301 [bis(2,4,4-trimethylpentyl) dithiophosphinic acid] and Cyanex 302 [bis(2,4,4-trimethylpentyl) monothiophosphinic acid]. The best separation results were obtained for extractants TBP and Cyanex 301. Around 92.5% of zinc and 11.2% of iron were extracted from effluent 1 in one single contact using 100% (v/v) of TBP. With Cyanex 301, around 80-95% of zinc and less than 10% of iron were extracted from effluent 2 at pH 0.3-1.0. For Cyanex 272, the highest extraction yield for zinc (70% of zinc with 20% of iron extraction) was found at pH 2.4. Cyanex 302 presented low metal extraction levels (below 10%) and slow phase disengagement characteristics. Reactions for the extraction of zinc with TBP and Cyanex 301 from hydrochloric acid solution were proposed.

  10. Optimization of squalene extraction from Palm Fatty Acid Distillate (PFAD) in multistage process

    NASA Astrophysics Data System (ADS)

    Sibuyo, Leah; Widiputri, Diah; Legowo, Evita

    2017-01-01

    Squalene is a compound widely known as one of the natural antioxidants used in the cosmetic and pharmaceutical industries. As the main source of squalene, which is shark liver oil, is becoming more limited in its availability, attempts have been made to extract squalene from other sources, e.g. from vegetable oils. Research has found that one of the wastes produced by palm oil industry, namely the palm fatty acid distillate (PFAD), contains squalene among other useful compounds. Since Indonesia is one of the largest producers of palm oil, the abundant amount of PFAD becomes very interesting to be a solution in coping with today demand of natural squalene. In this research, the extraction of squalene from PFAD is optimized through a multiple-stage extraction process, where results show a significant increase of squalene yield. Furthermore, the liquid-liquid phase equilibrium data for an extraction using dichloromethane (DCM) were plotted to develop a ternary-phase-diagram between squalene, DCM and free-fatty acids.

  11. Antioxidant activity, anti-proliferative activity, and amino acid profiles of ethanolic extracts of edible mushrooms.

    PubMed

    Panthong, S; Boonsathorn, N; Chuchawankul, S

    2016-10-17

    Biological activities of various mushrooms have recently been discovered, particularly, immunomodulatory and antitumor activities. Herein, three edible mushrooms, Auricularia auricula-judae (AA), Pleurotus abalonus (PA) and Pleurotus sajor-caju (PS) extracted using Soxhlet ethanol extraction were evaluated for their antioxidative, anti-proliferative effects on leukemia cells. Using the Folin-Ciocalteau method and Trolox equivalent antioxidant capacity assay, phenolics and antioxidant activity were found in all sample mushrooms. Additionally, anti-proliferative activity of mushroom extracts against U937 leukemia cells was determined using a viability assay based on mitochondrial activity. PA (0.5 mg/mL) and AA (0.25-0.5 mg/mL) significantly reduced cell viability. Interestingly, PS caused a hormetic-like biphasic dose-response. Low doses (0-0.25 mg/L) of PS promoted cell proliferation up to 140% relative to control, whereas higher doses (0.50 mg/mL) inhibited cell proliferation. Against U937 cells, AA IC50 was 0.28 ± 0.04 mg/mL, which was lower than PS or PA IC50 (0.45 ± 0.01 and 0.49 ± 0.001 mg/mL, respectively). Furthermore, lactate dehydrogenase (LDH) leakage conferred cytotoxicity. PS and PA were not toxic to U937 cells at any tested concentration; AA (0.50 mg/mL) showed high LDH levels and caused 50% cytotoxicity. Additionally, UPLC-HRMS data indicated several phytochemicals known to support functional activities as either antioxidant or anti-proliferative. Glutamic acid was uniquely found in ethanolic extracts of AA, and was considered an anti-cancer amino acid with potent anti-proliferative effects on U937 cells. Collectively, all mushroom extracts exhibited antioxidant effects, but their anti-proliferative effects were dose-dependent. Nevertheless, the AA extract, with highest potency, is a promising candidate for future applications.

  12. Effect of temperature on the extraction of uranium(VI) from nitric acid by tri-n-amyl phosphate

    SciTech Connect

    Srinivasan, T.G.; Rao, P.R.V.; Sood, D.D. |

    1997-01-01

    Studies have been carried out on the effect of temperature on the extraction of U(VI) from nitric acid medium by tri-n-amyl phosphate/n-dodecane, measured as a function of the extractant concentration and aqueous phase acidity. The results indicate that the extraction is exothermic as in the case of tri-n-butyl phosphate. From the data available an effort has been made to calculate the equilibrium constant, the Gibbs energy change and the entropy changes of the extraction reaction. 21 refs., 3 figs., 4 tabs.

  13. Suitability of Soxhlet extraction to quantify microalgal Fatty acids as determined by comparison with in situ transesterification.

    PubMed

    McNichol, Jesse; MacDougall, Karen M; Melanson, Jeremy E; McGinn, Patrick J

    2012-02-01

    To assess Soxhlet extraction as a method for quantifying fatty acids (FA) of microalgae, crude lipid, FA content from Soxhlet extracts and FA content from in situ transesterification (ISTE) were compared. In most cases, gravimetric lipid content was considerably greater (up to sevenfold) than the FA content of the crude lipid extract. FA content from Soxhlet lipid extraction and ISTE were similar in 12/18 samples, whereas in 6/18 samples, total FA content from Soxhlet extraction was less than the ISTE procedure. Re-extraction of residual biomass from Soxhlet extraction with ISTE liberated a quantity of FA equivalent to this discrepancy. Employing acid hydrolysis before Soxhlet extraction yielded FA content roughly equivalent to ISTE, indicating that acidic conditions of ISTE are responsible for this observed greater recovery of FA. While crude lipid derived from Soxhlet extraction was not a useful proxy for FA content for the species tested, it is effective in most strains at extracting total saponifiable lipid. Lipid class analysis showed the source of FA was primarily polar lipids in most samples (12/18 lipid extracts contained <5% TAG), even in cases where total FA content was high (>15%). This investigation confirms the usefulness of ISTE, reveals limitations of gravimetric methods for projecting biodiesel potential of microalgae, and reinforces the need for intelligent screening using both FA and lipid class analysis.

  14. Isolation and pharmacological characterization of fatty acids from saw palmetto extract.

    PubMed

    Abe, Masayuki; Ito, Yoshihiko; Suzuki, Asahi; Onoue, Satomi; Noguchi, Hiroshi; Yamada, Shizuo

    2009-04-01

    Saw palmetto extract (SPE) has been widely used for the treatment of lower urinary-tract symptoms secondary to benign prostatic hyperplasia. The mechanisms of pharmacological effects of SPE include the inhibition of 5alpha-reductase, anti-androgenic effects, anti-proliferative effects, and anti-inflammatory effects. Previously, we showed that SPE bound actively to alpha(1)-adrenergic, muscarinic and 1,4-dihydropyridine calcium channel (1,4-DHP) receptors in the prostate and bladder of rats, whereas its active constituents have not been fully clarified. The present investigation is aimed to identify the main active components contained in hexane and diethyl ether extracts of SPE with the use of column chromatography and preparative HPLC. Based on the binding activity with alpha(1)-adrenergic, muscarinic, and 1,4-DHP receptors, both isolated oleic and lauric acids were deduced to be active components. Authentic samples of oleic and lauric acids also exhibited similar binding activities to these receptors as the fatty acids isolated from SPE, consistent with our findings. In addition, oleic and lauric acids inhibited 5alpha-reductase, possibly leading to therapeutic effects against benign prostatic hyperplasia and related lower urinary-tract symptoms.

  15. Thallium extraction from hydrochloric acid media into a deep eutectic solvent using bis(2-ethylhexyl) phosphate

    NASA Astrophysics Data System (ADS)

    Tran, Kate; Volia, Merinda; Tereshatov, Evgeny; Folden, Charles, III

    2016-09-01

    The chemical properties of superheavy elements are relatively unknown due to their short half-lives and difficulty of production. In preparation for a future experiment to study the chemical properties of element 113, separation techniques have been used to study the behavior of its homologs, In and Tl. Previous work studied the liquid-liquid extraction of radioactive 201Tl (t1 / 2 = 3 . 04 d) from various concentrations of HCl into a mixture of menthol and lauric acid that formed a so-called deep eutectic solvent (DES). This work focuses on the effects of adding an extraction agent, bis(2-ethylhexyl) phosphate (HDEHP), to the DES on the efficiency of thallium extraction. The extraction of Tl(I) was generally poor, both with and without HDEHP added. In contrast, 111In (t1 / 2 = 2 . 80 d) showed significant extraction using HDEHP added to the same DES. This difference in behavior could potentially be exploited in a future experiment on the chemistry of element 113. National Science Foundation.

  16. Evaluation of Liquid-Liquid Extraction Process for Separating Acrylic Acid Produced From Renewable Sugars

    NASA Astrophysics Data System (ADS)

    Alvarez, M. E. T.; Moraes, E. B.; Machado, A. B.; Maciel Filho, R.; Wolf-Maciel, M. R.

    In this article, the separation and the purification of the acrylic acid produced from renewable sugars were studied using the liquid-liquid extraction process. Nonrandom two-liquids and universal quasi-chemical models and the prediction method univeral quasi-chemical functional activity coefficients were used for generating liquid-liquid equilibrium diagrams for systems made up of acrylic acid, water, and solvents (diisopropyl ether, isopropyl acetate, 2-ethyl hexanol, and methyl isobutyl ketone) and the results were compared with available liquid-liquid equilibrium experimental data. Aspen Plus (Aspen Technology, Inc., version 2004.1) software was used for equilibrium and process calculations. High concentration of acrylic acid was obtained in this article using diisopropyl ether as solvent.

  17. Evaluation of liquid-liquid extraction process for separating acrylic acid produced from renewable sugars.

    PubMed

    Alvarez, M E T; Moraes, E B; Machado, A B; Maciel Filho, R; Wolf-Maciel, M R

    2007-04-01

    In this article, the separation and the purification of the acrylic acid produced from renewable sugars were studied using the liquid-liquid extraction process. Nonrandom two-liquids and universal quasi-chemical models and the prediction method universal quasi-chemical functional activity coefficients were used for generating liquid-liquid equilibrium diagrams for systems made up of acrylic acid, water, and solvents (diisopropyl ether, isopropyl acetate, 2-ethyl hexanol, and methyl isobutyl ketone) and the results were compared with available liquid-liquid equilibrium experimental data. Aspen Plus (Aspen Technology, Inc., version 2004.1) software was used for equilibrium and process calculations. High concentration of acrylic acid was obtained in this article using diisopropyl ether as solvent.

  18. Selective and cost-effective protocol to separate bioactive triterpene acids from plant matrices using alkalinized ethanol: Application to leaves of Myrtaceae species

    PubMed Central

    Lima, Adélia M. Belem; Siani, Antonio Carlos; Nakamura, Marcos Jun; D’Avila, Luiz Antonio

    2015-01-01

    Background: Triterpenes as betulinic (BA), oleanolic (OA) and ursolic acids (UA) have increasingly gained therapeutic relevance due to their wide scope of pharmacological activities. To fit large-scale demands, exploitable sources of these compounds have to be found and simple, cost-effective methods to extract them developed. Leaf material represents the best plant sustainable raw material. To obtain triterpene acid-rich extracts from leaves of Eugenia, Psidium and Syzygium species (Myrtaceae) by directly treating the dry plant material with alkalinized hydrated ethanol. This procedure was adapted from earlier methods to effect depolymerization of the leaf cutin. Materials and Methods: Extracts were prepared by shaking the milled dry leaves in freshly prepared 2% NaOH in 95% EtOH solution (1:4 w/v) at room temperature for 6 h. Working up the product in acidic aqueous medium led to clear precipitates in which BA, OA and UA were quantified by gas chromatography. Results: Pigment-free and low-polyphenol content extracts (1.2–2.8%) containing 6–50% of total triterpene acids were obtained for the six species assayed. UA (7–20%) predominated in most extracts, but BA preponderated in Eugenia florida (39%). Carried out in parallel, n-hexane defatted leaves led to up to 9% enhancement of total acids in the extracts. The hydroalcoholate treatment of Myrtaceae species dry leaves proved to be a cost-effective and environmentally friendly method to obtain triterpene acids, providing them be resistant to alkaline medium. These combined techniques might be applicable to other plant species and tissues. PMID:26246721

  19. Extraction of Am(III), Eu(III) and U(VI) from perchloric acid and mixtures of acids by dialkyl-(diaryl) [diethylcarbamoylmethyl] phosphine oxides

    SciTech Connect

    Chmutova, M.K.; Litvina, M.N.; Nesterova, N.P.; Myasoedov, B.F.; Kabachnik, M.I.

    1992-07-01

    Extraction of Am(III), and Eu(III) and U(VI) from perchloric acid solutions by dialkyl(diaryl) [diethyl-carbamoylmethyl] phosphine oxides (CMPO) has been investigated. It has been shown that elements are extracted more effectively from perchloric acid solutions than from nitric acid. An `anomalous aryl effect` increases sufficiently in perchloric media as compared with nitric media. Solvate numbers of elements also increase in perchloric acid, they being extremely high for aryl-substituted reagents and not so high for alkyl-substituted compounds. It has been shown that in HNO{sub 3} and HClO{sub 4} solutions the value of `anomalous aryl effect` directly depends on the difference in values of element solvate numbers in complexes with aryl- and alkyl-substituted CMPO. Am(III), Eu(III) and U(VI) are not practically extracted from phosphoric and sulphuric acids by CMPO solutions. 16 refs., 7 figs., 3 tabs.

  20. Comparison of the Bruker Biotyper and Vitek MS matrix-assisted laser desorption ionization-time of flight mass spectrometry systems for identification of mycobacteria using simplified protein extraction protocols.

    PubMed

    Mather, Cheryl A; Rivera, Sheila F; Butler-Wu, Susan M

    2014-01-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has recently been described as a fast and inexpensive method for the identification of mycobacteria. Although mycobacteria require extraction prior to MALDI-TOF MS analysis, previously published protocols have been relatively complex, involving significant hands-on time and materials not often found in the clinical laboratory. In this study, we tested two simplified protein extraction protocols developed at the University of Washington (UW) and by bioMérieux (BMX) for use with two different mass spectrometry platforms (the Bruker MALDI Biotyper and the bioMérieux Vitek MS, respectively). Both extraction protocols included vortexing with silica beads in the presence of ethanol. The commercial Bruker database was also augmented with an in-house database composed of 123 clinical Mycobacterium strains. A total of 198 clinical strains, representing 18 Mycobacterium species, were correctly identified to the species level 94.9% of the time when extracted using the UW protocol and compared to the augmented database. The BMX protocol and Vitek MS system resulted in correct species-level identifications for 94.4% of these strains. In contrast, only 79.3% of the strains were identified to the species level by the nonaugmented Bruker database, although the use of a lower identification score threshold (≥1.7) increased the identification rate to 93.9%, with two misidentifications that were unlikely to be clinically relevant. The two simplified protein extraction protocols described in this study are easy to use for identifying commonly encountered Mycobacterium species.

  1. Antinutritional factor content and hydrochloric acid extractability of minerals in pearl millet cultivars as affected by germination.

    PubMed

    Abdelrahaman, Samia M; Elmaki, Hagir B; Idris, Wisal H; Hassan, Amro B; Babiker, Elfadil E; El Tinay, Abdullahi H

    2007-02-01

    Four pearl millet cultivars of two different species--Kordofani and Ugandi (Pennisetum typhoideum) and Madelkawaya and Shambat (Pennisetum glaucum)--were germinated for 6 days. The germinated grains were dried and milled. Phytic acid and polyphenol contents and hydrochloric acid (HCl) extractability of minerals from the malt flours were determined at intervals of 2 days during germination. Phytic acid and polyphenol contents decreased significantly (P <0.01) with an increase in germination time, with a concomitant increase in HCl extractable minerals. However, the major mineral content was significantly decreased while that of trace minerals was increased with germination time. When the grains were germinated for 6 days, Madelkawaya had higher extractable calcium while Ugandi had higher extractable phosphorus, whereas iron and manganese recorded high levels in Shambat and Madelkawaya, respectively. There was good correlation between antinutritional factors reduction and the increment in extractable minerals with germination time.

  2. Vine-shoot waste aqueous extract applied as foliar fertilizer to grapevines: Effect on amino acids and fermentative volatile content.

    PubMed

    Sánchez-Gómez, R; Garde-Cerdán, T; Zalacain, A; Garcia, R; Cabrita, M J; Salinas, M R

    2016-04-15

    The aim of this work was to study the influence of foliar applications of different wood aqueous extracts on the amino acid content of musts and wines from Airén variety; and to study their relationship with the volatile compounds formed during alcoholic fermentation. For this purpose, the foliar treatments proposed were a vine-shoot aqueous extract applied in one and two times, and an oak extract which was only applied once. Results obtained show the potential of Airén vine-shoot waste aqueous extracts to be used as foliar fertilizer, enhancing the wine amino acid content especially when they were applied once. Similar results were observed with the aqueous oak extract. Regarding wine fermentative volatile compounds, there is a close relationship between musts and their wines amino acid content allowing us to discuss about the role of proline during the alcoholic fermentation and the generation of certain volatiles.

  3. Extraction of uranium: comparison of stripping with ammonia vs. strong acid

    SciTech Connect

    Moldovan, B.; Grinbaum, B.; Efraim, A.

    2008-07-01

    Following extraction of uranium in the first stage of solvent extraction using a tertiary amine, typically Alamine 336, the stripping of the extracted uranium is accomplished either by use of an aqueous solution of (NH{sub 4}){sub 2}SO{sub 4} /NH{sub 4}OH or by strong-acid stripping using 400-500 g/L H{sub 2}SO{sub 4}. Both processes have their merits and determine the downstream processing. The classical stripping with ammonia is followed by addition of strong base, to precipitate ammonium uranyl sulfate (NH{sub 4}){sub 2}UO{sub 2}(SO{sub 4}){sub 2}, which yields finally the yellow cake. Conversely, stripping with H{sub 2}SO{sub 4}, followed by oxidation with hydrogen peroxide yields uranyl oxide as product. At the Cameco Key Lake operation, both processes were tested on a pilot scale, using a Bateman Pulsed Column (BPC). The BPC proved to be applicable to both processes. It met the process criteria both for extraction and stripping, leaving less than 1 mg/L of U{sub 3}O{sub 8} in the raffinate, and product solution had the required concentration of U{sub 3}O{sub 8} at high flux and reasonable height of transfer unit. In the Key Lake mill, each operation can be carried out in a single column. The main advantages of the strong-acid stripping over ammonia stripping are: (1) 60% higher flux in the extraction, (2) tenfold higher concentration of the uranium in the product solution, and (3) far more robust process, with no need of pH control in the stripping and no need to add acid to the extraction in order to keep the pH above the point of precipitation of iron compounds. The advantages of the ammoniacal process are easier stripping, that is, less stages needed to reach equilibrium and lower concentration of modifier needed to prevent the creation of a third phase. (authors)

  4. Microbial process for the preparation of acetic acid as well as solvent for its extraction from the fermentation broth

    DOEpatents

    Gaddy, James L.; Clausen, Edgar C.; Ko, Ching-Whan; Wade, Leslie E.; Wikstrom, Carl V.

    2006-07-11

    A modified water-immiscible solvent useful in the extraction of acetic acid from aqueous streams is a substantially pure mixture of isomers of highly branched di-alkyl amines. This solvent is substantially devoid of mono-alkyl amines and alcohols. Solvent mixtures formed of such a modified solvent with a desired cosolvent, preferably a low boiling hydrocarbon which forms an azeotrope with water are useful in the extraction of acetic acid from aqueous gaseous streams. An anaerobic microbial fermentation process for the production of acetic acid employs such solvents, under conditions which limit amide formation by the solvent and thus increase the efficiency of acetic acid recovery. Methods for the direct extraction of acetic acid and the extractive fermentation of acetic acid also employ the modified solvents and increase efficiency of acetic acid production. Such increases in efficiency are also obtained where the energy source for the microbial fermentation contains carbon dioxide and the method includes a carbon dioxide stripping step prior to extraction of acetic acid in solvent.

  5. Microbial process for the preparation of acetic acid as well as solvent for its extraction from the fermentation broth

    DOEpatents

    Gaddy, James L.; Clausen, Edgar C.; Ko, Ching-Whan; Wade, Leslie E.; Wikstrom, Carl V.

    2002-01-01

    A modified water-immiscible solvent useful in the extraction of acetic acid from aqueous streams is a substantially pure mixture of isomers of highly branched di-alkyl amines. This solvent is substantially devoid of mono-alkyl amines and alcohols. Solvent mixtures formed of such a modified solvent with a desired cosolvent, preferably a low boiling hydrocarbon which forms an azeotrope with water are useful in the extraction of acetic acid from aqueous gaseous streams. An anaerobic microbial fermentation process for the production of acetic acid employs such solvents, under conditions which limit amide formation by the solvent and thus increase the efficiency of acetic acid recovery. Methods for the direct extraction of acetic acid and the extractive fermentation of acetic acid also employ the modified solvents and increase efficiency of acetic acid production. Such increases in efficiency are also obtained where the energy source for the microbial fermentation contains carbon dioxide and the method includes a carbon dioxide stripping step prior to extraction of acetic acid in solvent.

  6. Controllable Phase Separation by Boc-Modified Lipophilic Acid as a Multifunctional Extractant

    NASA Astrophysics Data System (ADS)

    Tao, Kai; Adler-Abramovich, Lihi; Gazit, Ehud

    2015-12-01

    While phase separation of immiscible liquid-liquid systems has become increasingly significant in diverse areas, the irreversible nature limits their further application in controllable extraction-concentration or capture-release fields. There is a need for the development of simple, efficient and reversible methods for numerous research and industrial extraction and separation applications. We envisioned Boc-modified lipophilic acids as a simple model for such use based on the studies of the multi-phase transitions of Boc-modified supramolecular polymeric systems. Here, we demonstrate that in the presence of Boc-7-aminoheptanoic acid (Boc-7), phase separation occurs in mixtures of miscible organic solvent and water. The separation behavior was confirmed by differential colorimetric development in aqueous and organic phases using methyl orange staining assays. Component substitution experiments verified that the phase separation results from the subtle balance between the aggregation and the solvation forces of Boc-7, and is reversible by adjusting the solution pH. Owing to the intrinsic hydrophobic properties of the organic phase and the hydrogen bonding-forming ability of the carboxyl group of Boc-7, the phase separation system captures and releases Sudan Red, fluorescein, and streptavidin in a controllable manner. Consequently, a reversible and simple phase separation system can be designed as a multifunctional extractant.

  7. Citric acid production from Aspergillus niger MT-4 using hydrolysate extract of the insect Locusta migratoria.

    PubMed

    Taskin, Mesut; Tasar, Gani Erhan; Incekara, Umit

    2013-06-01

    Citric acid (CA) is the most important organic acid used in the food and other industries. Locusta migratoria is an insect species, which has rich nutritional composition (especially protein) and cultivated in some countries. Therefore, the present study investigated the usability of hydrolysate extract of L. migratoria biomass as substrate for the production of CA from Aspergillus niger MT-4. The insect extract (IE) was found to be rich in ash (34.9 g/100 g), protein (35.6 g/100 g) and mineral contents. Yeast extract was found to be the most favorable substrate for biomass production, whereas the maximum production of CA (41.8 g/L) was achieved in the medium containing IE. Besides, uniform pellets with the smallest size (4 mm) were observed in IE medium. It was thought that rich magnesium (6.78 g/100 g) and manganese (1.14 g/100 g) contents of IE increased the production of CA, resulting in the formation of small uniform pellets. This is the first report on the effect of protein-rich insect biomasses on the production of CA. In this regard, L. migratoria biomass was tested for the first time as a CA-production substrate.

  8. Controllable Phase Separation by Boc-Modified Lipophilic Acid as a Multifunctional Extractant

    PubMed Central

    Tao, Kai; Adler-Abramovich, Lihi; Gazit, Ehud

    2015-01-01

    While phase separation of immiscible liquid-liquid systems has become increasingly significant in diverse areas, the irreversible nature limits their further application in controllable extraction-concentration or capture-release fields. There is a need for the development of simple, efficient and reversible methods for numerous research and industrial extraction and separation applications. We envisioned Boc-modified lipophilic acids as a simple model for such use based on the studies of the multi-phase transitions of Boc-modified supramolecular polymeric systems. Here, we demonstrate that in the presence of Boc-7-aminoheptanoic acid (Boc-7), phase separation occurs in mixtures of miscible organic solvent and water. The separation behavior was confirmed by differential colorimetric development in aqueous and organic phases using methyl orange staining assays. Component substitution experiments verified that the phase separation results from the subtle balance between the aggregation and the solvation forces of Boc-7, and is reversible by adjusting the solution pH. Owing to the intrinsic hydrophobic properties of the organic phase and the hydrogen bonding-forming ability of the carboxyl group of Boc-7, the phase separation system captures and releases Sudan Red, fluorescein, and streptavidin in a controllable manner. Consequently, a reversible and simple phase separation system can be designed as a multifunctional extractant. PMID:26627307

  9. Solvent extraction of scandium from malonic acid with high molecular-weight amines.

    PubMed

    Dalvi, M B; Khopkar, S M

    1979-09-01

    Scandium is quantitatively extracted with 4% Amberlite LA-1 or Amberlite LA-2 in xylene at pH 2.5-5.5 from 0.1M malonic acid. Scandium is stripped from the organic phase with 0.5M hydrochloric acid and determined spectrophotometrically at 525 nm, as its complex with Alizarin Red S. Primene JM-T, tri-iso-octylamine, tributylamine and tribenzylamine have also been studied as extractants, but found to be unsatisfactory for various reasons. Xylene, toluene, benzene, chloroform, carbon tetrachloride, hexane, cyclohexane and kerosene have been studied as diluents. Xylene is found to be the most efficient. Scandium can be separated from most metals by selective extraction, and from gallium, thallium(III), bismuth, antimony(III), chromium(III), copper(II), iron(III), uranium(VI), cerium, zirconium, indium, thorium and titanium by selective stripping, in some cases combined with use of suitable complexing media to retain the other metals in the organic phase.

  10. Use of lanthanum and sulphuric acid to suppress interferences in the flame photometric determination of calcium m soil extracts.

    PubMed

    Evans, C C; Grimshaw, H M

    1968-04-01

    Interference by iron, aluminium and phosphate in the flame photometric determination of calcium in soil extracts is not fully suppressed by lanthanum unless dilute sulphuric acid is also present. The investigation was restricted to the oxy-acetylene flame.

  11. Freeze-out extraction of monocarboxylic acids from water into acetonitrile under the action of centrifugal forces

    NASA Astrophysics Data System (ADS)

    Bekhterev, V. N.

    2016-10-01

    It is established that the efficiency of the freezing-out extraction of monocarboxylic acids C3-C;8 and sorbic acid from water into acetonitrile increases under the action of centrifugal forces. The linear growth of the partition coefficient in the homologous series of C2-C8 acids with an increase in molecule length, and the difference between the efficiency of extracting sorbic and hexanoic acid, are discussed using a theoretical model proposed earlier and based on the adsorption-desorption equilibrium of the partition of dissolved organic compounds between the resulting surface of ice and the liquid phase of the extract. The advantages of the proposed technique with respect to the degree of concentration over the method of low-temperature liquid-liquid extraction are explained in light of the phase diagram for the water-acetonitrile mixture.

  12. Revisiting the ALA/N (alpha-lipoic acid/low-dose naltrexone) protocol for people with metastatic and nonmetastatic pancreatic cancer: a report of 3 new cases.

    PubMed

    Berkson, Burton M; Rubin, Daniel M; Berkson, Arthur J

    2009-12-01

    The authors, in a previous article, described the long-term survival of a man with pancreatic cancer and metastases to the liver, treated with intravenous alpha-lipoic acid and oral low-dose naltrexone (ALA/N) without any adverse effects. He is alive and well 78 months after initial presentation. Three additional pancreatic cancer case studies are presented in this article. At the time of this writing, the first patient, GB, is alive and well 39 months after presenting with adenocarcinoma of the pancreas with metastases to the liver. The second patient, JK, who presented to the clinic with the same diagnosis was treated with the ALA/N protocol and after 5 months of therapy, PET scan demonstrated no evidence of disease. The third patient, RC, in addition to his pancreatic cancer with liver and retroperitoneal metastases, has a history of B-cell lymphoma and prostate adenocarcinoma. After 4 months of the ALA/N protocol his PET scan demonstrated no signs of cancer. In this article, the authors discuss the poly activity of ALA: as an agent that reduces oxidative stress, its ability to stabilize NF(k)B, its ability to stimulate pro-oxidant apoptosic activity, and its discriminative ability to discourage the proliferation of malignant cells. In addition, the ability of lowdose naltrexone to modulate an endogenous immune response is discussed. This is the second article published on the ALA/N protocol and the authors believe the protocol warrants clinical trial.

  13. Citric acid production from extract of Jerusalem artichoke tubers by the genetically engineered yeast Yarrowia lipolytica strain 30 and purification of citric acid.

    PubMed

    Wang, Ling-Fei; Wang, Zhi-Peng; Liu, Xiao-Yan; Chi, Zhen-Ming

    2013-11-01

    In this study, citric acid production from extract of Jerusalem artichoke tubers by the genetically engineered yeast Yarrowia lipolytica strain 30 was investigated. After the compositions of the extract of Jerusalem artichoke tubers for citric acid production were optimized, the results showed that natural components of extract of Jerusalem artichoke tubers without addition of any other components were suitable for citric acid production by the yeast strain. During 10 L fermentation using the extract containing 84.3 g L(-1) total sugars, 68.3 g L(-1) citric acid was produced and the yield of citric acid was 0.91 g g(-1) within 336 h. At the end of the fermentation, 9.2 g L(-1) of residual total sugar and 2.1 g L(-1) of reducing sugar were left in the fermented medium. At the same time, citric acid in the supernatant of the culture was purified. It was found that 67.2 % of the citric acid in the supernatant of the culture was recovered and purity of citric acid in the crystal was 96 %.

  14. Exhaustive and stable electromembrane extraction of acidic drugs from human plasma.

    PubMed

    Huang, Chuixiu; Gjelstad, Astrid; Seip, Knut Fredrik; Jensen, Henrik; Pedersen-Bjergaard, Stig

    2015-12-18

    The first part of the current work systematically described the screening of different types of organic solvents as the supported liquid membrane (SLM) for electromembrane extraction (EME) of acidic drugs, including different alcohols, ketones, and ethers. Seven acidic drugs with a wide logP range (1.01-4.39) were selected as model substances. For the first time, the EME recovery of acidic drugs and system-current across the SLM with each organic solvent as SLM were investigated and correlated to relevant solvent properties such as viscosity and Kamlet and Taft solvatochromic parameters. Solvents with high hydrogen bonding acidity (α) and dipolarity-polarizability (π*) were found to be successful SLMs, and 1-heptanol was the most efficient candidate, which provided EME recovery in the range of 94-110%. Both hydrogen bonding interactions, dipole-dipole interactions, and hydrophobic interactions were involved in stabilizing the deprotonated acidic analytes (with high hydrogen bonding basicity and high dipole moment) during mass transfer across the SLM. The efficiency of the extraction normally decreased with increasing hydrocarbon chain length of the SLM, which was mainly due to increasing viscosity and decreasing α and π* values. The system-current during EME was found to be dependent on the type and the volume of the SLM. In contact with human plasma, an SLM of pure 1-heptanol was unstable, and to improve stability, 1-heptanol was mixed with 2-nitrophenyl octyl ether (NPOE). With this SLM, exhaustive EME was performed from diluted human plasma, and the recoveries of five out of seven analytes were over 91% after 10min EME. This approach was evaluated using HPLC-UV, and the evaluation data were found to be satisfactory.

  15. Development of a solid-phase extraction/gas chromatographic-mass spectrometric method for quantification of succinic acid in nucleoside derivatives for oligonucleotide synthesis.

    PubMed

    Stenholm, Ake; Drevin, Ingrid; Lundgren, Michael

    2005-04-08

    A solid-phase extraction (SPE)/gas chromatographic-mass spectrometric (GC-MS) method was developed for analysing residual succinic acid in nucleoside derivatives to be used in oligonucleotide synthesis. Use of a SPE protocol, enabled most of the derivatives to be trapped, thereby creating eluates enriched in succinic acid. GC-MS was used to quantify the amount of residual succinic acid in four different nucleoside preparations, with succinate concentrations varying from 0.18 to 0.24% (w/w). The within-day repeatability of the method was found to be 1.25% RSD. A linear relationship was observed between the amount of succinic acid in the sample and the GC-MS peak area, with a correlation coefficient of 0.9997 in the concentration interval 0.05-2.5% (w/w). Recoveries were measured by the addition of internal standards to working solutions and varied between 99.8 and 102.6%.

  16. In situ extractive fermentation for the production of hexanoic acid from galactitol by Clostridium sp. BS-1.

    PubMed

    Jeon, Byoung Seung; Moon, Chuloo; Kim, Byung-Chun; Kim, Hyunook; Um, Youngsoon; Sang, Byoung-In

    2013-08-15

    Clostridium sp. BS-1 produces hexanoic acid as a metabolite using galactitol and enhanced hexanoic acid production was obtained by in situ extractive fermentation with Clostridium sp. BS-1 under an optimized medium composition. For medium optimization, five ingredients were selected as variables, and among them yeast extract, tryptone, and sodium butyrate were selected as significant variables according to a fractional factorial experimental design, a steepest ascent experimental design, and a Box-Behnken experimental design. The optimized medium had the following compositions in modified Clostridium acetobutyricum (mCAB) medium: 15.5gL(-1) of yeast extract, 10.13gL(-1) of tryptone, 0.04gL(-1) of FeSO4·7H2O, 0.85gL(-1) of sodium acetate, and 6.47gL(-1) of sodium butyrate. The predicted concentration of hexanoic acid with the optimized medium was 6.98gL(-1), and this was validated experimentally by producing 6.96gL(-1) of hexanoic acid with Clostridium sp. BS-1 under the optimized conditions. In situ extractive fermentation for hexanoic acid removal was then applied in a batch culture system with the optimized medium and 10% (v/v) alamine 336 in oleyl alcohol as an extractive solvent. The pH of the culture in the extractive fermentation was maintained at 5.4-5.6 by an acid balance between production and retrieval by extraction. During a 16 day culture, the hexanoic acid concentration in the solvent increased to 32gL(-1) while it was maintained in a range of 1-2gL(-1) in the medium. The maximum rate of hexanoic acid production was 0.34gL(-1)h(-1) in in situ extractive fermentation.

  17. Cleaner production of citric acid by recycling its extraction wastewater treated with anaerobic digestion and electrodialysis in an integrated citric acid-methane production process.

    PubMed

    Xu, Jian; Su, Xian-Feng; Bao, Jia-Wei; Chen, Yang-Qiu; Zhang, Hong-Jian; Tang, Lei; Wang, Ke; Zhang, Jian-Hua; Chen, Xu-Sheng; Mao, Zhong-Gui

    2015-01-01

    To solve the pollution problem of extraction wastewater in citric acid production, an integrated citric acid-methane production process was proposed. Extraction wastewater was treated through anaerobic digestion and the anaerobic digestion effluent (ADE) was recycled for the next batch of citric acid fermentation, thus eliminating wastewater discharge and reducing water consumption. Excessive Na(+) contained in ADE could significantly inhibit citric acid fermentation in recycling and was removed by electrodialysis in this paper. Electrodialysis performance was improved after pretreatment of ADE with air stripping and activated carbon adsorption to remove precipitable metal ions and pigments. Moreover, the concentrate water was recycled and mixed with feed to improve the water recovery rate above 95% in electrodialysis treatment, while the dilute water was collected for citric acid fermentation. The removal rate of Na(+) in ADE was above 95% and the citric acid production was even higher than that with tap water.

  18. Carob pod water extracts as feedstock for succinic acid production by Actinobacillus succinogenes 130Z.

    PubMed

    Carvalho, Margarida; Roca, Christophe; Reis, Maria A M

    2014-10-01

    Carob pods are a by-product of locust bean gum industry containing more than 50% (w/w) sucrose, glucose and fructose. In this work, carob pod water extracts were used, for the first time, for succinic acid production by Actinobacillus succinogenes 130Z. Kinetic studies of glucose, fructose and sucrose consumption as individual carbon sources till 30g/L showed no inhibition on cell growth, sugar consumption and SA production rates. Sugar extraction from carob pods was optimized varying solid/liquid ratio and extraction time, maximizing sugar recovery while minimizing the extraction of polyphenols. Batch fermentations containing 10-15g/L total sugars resulted in a maximum specific SA production rate of 0.61Cmol/Cmol X.h, with a yield of 0.55Cmol SA/Cmol sugar and a volumetric productivity of 1.61g SA/L.h. Results demonstrate that carob pods can be a promising low cost feedstock for bio-based SA production.

  19. Cloud point extraction of uranium using H₂DEH[MDP] in acidic conditions.

    PubMed

    Labrecque, Charles; Potvin, Sabrina; Whitty-Léveillé, Laurence; Larivière, Dominic

    2013-03-30

    A procedure has been developed for the cloud point extraction (CPE) of uranium (VI) using H2DEH[MDP] (P,P-di(2-ethylhexyl) methanediphosphonic acid) with inductively coupled plasma coupled to mass spectrometry (ICP-MS). The method is based on the modification of the cloud point temperature using cetyl trimethyl ammonium bromide (CTAB) and KI. Optimal conditions of extraction were found using a cross-optimization of every parameter (non-ionic and ionic surfactant concentrations, chelating agent concentration, pH and the extraction, and phase separation temperatures). Furthermore, the figures of merit of the methodology were assessed (limit of detection, limit of quantification, recovery, sensibility, and linear range) and are reported. Quantitative extraction (99 ± 0.5%) was obtained in drinking water samples over a wide range of uranium concentrations. The approach was also validated using drinking (SCP EP-L-3 and SCP EP-H-3), and wastewater (SCP EU-L-3) certified materials. Interferences from most critical anions and cations were evaluated to determine the reliability of the method. The proposed method showed robustness since its performance is maintained over a wide range of pH and metal ion concentrations.

  20. Combination of graphene oxide-based solid phase extraction and electro membrane extraction for the preconcentration of chlorophenoxy acid herbicides in environmental samples.

    PubMed

    Tabani, Hadi; Fakhari, Ali Reza; Shahsavani, Abolfath; Behbahani, Mohammad; Salarian, Mani; Bagheri, Akbar; Nojavan, Saeed

    2013-07-26

    Combination of different extraction methods is an interesting and debatable work in the field of sample preparation. In the current study, for the first time, solid phase extraction combined with electro membrane extraction (SPE-EME) was developed for ultra-preconcentration and determination of chlorophenoxy acid herbicides in environmental samples using capillary electrophoresis (CE). In the mentioned method, first, a 100mL of chlorophenoxy acid herbicides (2-methyl-4-chlorophenoxyacetic acid (MCPA), 2-(2,4-dichlorophenoxy) propanoic acid (2,4-DP) and 2-(4-chloro-2-methylphenoxy) propanoic acid (MCPP)) was passed through a column of graphene oxide as a solid phase, and then the adsorbed herbicides were eluted by 4.0mL of 8% acetic acid (HOAC) in methanol. Then, the elution solvent was evaporated and the herbicides residue was dissolved in 4.0mL of double distilled water (pH 9.0). Afterwards, the herbicides in 4.0mL of the aqueous solution were transferred to an EME glass vial. In the EME step, the herbicides were extracted from the sample solution into the basic acceptor solution (pH 13.0) under electrical potential, which was held inside the lumen of the fiber with 1-octanol as the supported liquid membrane (SLM). Under the optimized conditions, high enrichment factors were obtained in the range of 1950-2000. The limits of quantification (LOQs) and method detection limits (MDLs) were obtained in the range of 1.0-1.5 and 0.3-0.5ngmL(-1), respectively. Finally, the performance of the present method was evaluated for extraction and determination of chlorophenoxy acid herbicides in environmental samples.

  1. Modulation of the immune system by Boswellia serrata extracts and boswellic acids.

    PubMed

    Ammon, H P T

    2010-09-01

    Extracts from the gum resin of Boswellia serrata and some of is constituents including boswellic acids affect the immune system in different ways. Among the various boswellic acids 11-keto-beta-boswellic acid (KBA) and acetyl-11-keto-beta-boswellic acid have been observed to be active. However, also other boswellic acids may exhibit actions in the immune system. In the humoral defence system a mixture of boswellic acis at higher doses reduced primary antibody titres; on the other hand lower doses enhanced secondary antibody titres following treatment with sheep erythrocytes. In the cellular defence boswellic acides appear to increase lymphocyte proliferation whereas higher concentrations are even inhibitory. Moreover, BAs increase phagocytosis of macrophages. BAs affect the cellular defence system by interaction with production/release of cytokines. Thus, BAs inhibit activation of NFkappaB which is a product of neutrophile granulocytes. Consequently a down regulation of TNF-alpha and decrease of IL-1, IL-2, IL-4, IL-6 and IFN-gamma, which are proinflammatory cytokines by BEs and BAs has been reported. Suppressions of the classic way of the complement system was found to be due to inhibition of the conversion of C3 into C3a and C3b. However, which of these pharmacological actions contribute to the therapeutic effects and which is finally the best dosage of a standardized extract needs further examination. And it is also a question whether or not a single BA will have the same therapeutic effect as a standardized extract. Among the mediators of inflammatory reaction, mast cell stabilisation has been described by a BE. Inhibition of prostaglandin synthesis appears to play only a minor role as far as the anti-inflammatory effect is concerned. On the other hand the inhibitory action of BAs on 5-LO leading to a decreased production of leukotrienes has received high attention by the scientific community since a variety of chronic inflammatory diseases is associatied with

  2. A Magnetic Nanoparticle Based Nucleic Acid Isolation and Purification Instrument for DNA Extraction of Escherichia Coli O157: H7.

    PubMed

    Chen, Yahui; Lin, Jianhan; Jiang, Qin; Chen, Qi; Zhang, Shengjun; Li, Li

    2016-03-01

    The objective of this study was to evaluate the performance of a nucleic acid isolation and purification instrument using Escherichia coli O157:H7 as the model. The instrument was developed with magnetic nanoparticles for efficiently capturing nucleic acids and an intelligent mechanical unit for automatically performing the whole nucleic acid extraction process. A commercial DNA extraction kit from Huier Nano Company was used as reference. Nucleic acids in 1 ml of E. coli O157: H7 at a concentration of 5 x 10(8) CFU/mL were extracted by using this instrument and the kit in parallel and then detected by an ultraviolet spectrophotometer to obtain A260 values and A260/A280 values for the determination of the extracted DNA's quantity and purity, respectively. The A260 values for the instrument and the kit were 0.78 and 0.61, respectively, and the A260/A280 values were 1.98 and 1.93. The coefficient of variations of these parallel tests ranged from 10.5% to 16.7%. The results indicated that this nucleic acid isolation and purification instrument could extract a comparable level of nucleic acid within 50 min compared to the commercial DNA extraction kit.

  3. Substituted ethan phosphonic acid esters as reagents for the separation of molybdenum from rhenium by solvent extraction.

    PubMed

    Jordanov, N; Mareva, S; Borisov, G; Jordanov, B

    1968-02-01

    New esters of the ethan phosphonic acids have been synthesized and their extraction properties studied with respect to Mo(VI), Re(VII), Fe(III),Au(III),Tl(III) and Sb(V). A possibility is shown for the analytical separation of molybdenum from rhenium (beta = 700). The state of these new extraction agents in carbon tetrachloride solution, and the mechanism of the extraction processes, have been investigated by means of infrared spectra.

  4. Extraction of formic and acetic acids from aqueous solution by dynamic headspace-needle trap extraction temperature and pH optimization.

    PubMed

    Lou, Da-Wei; Lee, Xinqing; Pawliszyn, Janusz

    2008-08-08

    A combined method of dynamic headspace-needle trap sample preparation and gas chromatography for the determination of formic and acetic acids in aqueous solution was developed in this study. A needle extraction device coupled with a gas aspirating pump was intended to perform sampling and preconcentration of target compounds from aqueous sample before gas chromatographic analysis. The needle trap extraction (NTE) technique allows for the successful sampling of short chain fatty acids under dynamic conditions while keeping the headspace (HS) volume constant. Two important parameters, including extraction temperature and effect of acidification, have been optimized and evaluated using the needle trap device. The method detection limits for the compounds estimated were 87.2microg/L for acetic acid and 234.8microg/L for formic acid in spite of the low flame ionization detection response for formic acid and its low Henry's law constant in aqueous solution. Precision was determined based on the two real samples and ranged between 4.7 and 10.7%. The validated headspace-needle trap extraction method was also successfully applied to several environmental samples.

  5. Evaluation of different solvent extraction methods for removing actinides from high acid waste streams

    SciTech Connect

    Yarbro, S.L.; Schreiber, S.B.; Dunn, S.L. ); Rogers, J. )

    1991-01-01

    At the Los Alamos National Laboratory Plutonium Facility, anion exchange is used to recover plutonium from nitric acid solutions. Although this approach recovers >99%, trace amounts of plutonium and other actinides remain the effluent and require additional processing. Currently, a ferric hydroxide carrier precipitation is used to remove the trace actinides and the resulting sludge is cemented. Because it costs approximately $10,000 per drum for disposal, we are developing an additional polishing step so that the effluent actinide levels are reduced to below 100 nCi/g. This would allow the resulting waste sludge to disposed as low-level waste at approximately $200 per drum. We are investigating various solvent extraction techniques for removing actinides. The most promising are chelating resins and membrane-based liquid-liquid solvent extraction. This report details some of our preliminary results. 4 refs., 3 tabs.

  6. Determination of chloroacetic acids in drinking water using suppressed ion chromatography with solid-phase extraction.

    PubMed

    Yoshikawa, Kenji; Soda, Yuko; Sakuragawa, Akio

    2009-12-01

    Suppressed ion chromatography with a conductivity detector was developed for the determination of trace amounts of underivatized chloroacetic acids (CAAs). When sodium carbonate and methanol were used as a mobile phase, the simultaneous determination of each CAA took approximately 25 min. The linearity, reproducibility and detection limits were determined for the proposed method. For the solid-phase extraction step, the effects of the pH of the sample solution, sample volume and the eluting agent were tested. Under the optimized extracting conditions, the average recoveries for CAAs spiked in tap water were 83-107%, with an optimal preconcentration factor of 20. The reproducibility of recovery rate for CAAs was 1.2-3.8%, based upon 6 repetitions of the recovery experiments.

  7. Simultaneous extraction and HPLC determination of 3-indole butyric acid and 3-indole acetic acid in pea plant by using ionic liquid-modified silica as sorbent.

    PubMed

    Sheikhian, Leila; Bina, Sedigheh

    2016-01-15

    In this study, ionic liquid-modified silica was used as sorbent for simultaneous extraction and preconcentration of 3-indole butyric acid and 3-indole acetic acid in pea plants. The effect of some parameters such as pH and ionic strength of sample solution, amount of sorbent, flow rate of aqueous sample solution and eluent solution, concentration of eluent solution, and temperature were studied for each hormone solution. Percent extraction of 3-indole butyric acid and 3-indole acetic acid was strongly affected by pH of aqueous sample solution. Ionic strength of aqueous phase and temperature showed no serious effects on extraction efficiency of studied plant hormones. Obtained breakthrough volume was 200mL for each of studied hormones. Preconcentration factor for spectroscopic and chromatographic determination of studied hormones was 100 and 4.0×10(3) respectively. Each solid sorbent phase was reusable for almost 10 times of extraction/stripping procedure. Relative standard deviations of extraction/stripping processes of 3-indole butyric acid and 3-indole acetic acid were 2.79% and 3.66% respectively. The calculated limit of detections for IBA and IAA were 9.1×10(-2)mgL(-1) and 1.6×10(-1)mgL(-1) respectively.

  8. Design and Performance Testing of a DNA Extraction Assay for Sensitive and Reliable Quantification of Acetic Acid Bacteria Directly in Red Wine Using Real Time PCR

    PubMed Central

    Longin, Cédric; Guilloux-Benatier, Michèle; Alexandre, Hervé

    2016-01-01

    Although strategies exist to prevent AAB contamination, the increased interest for wines with low sulfite addition leads to greater AAB spoilage. Hence, there is a real need for a rapid, specific, sensitive, and reliable method for detecting these spoilage bacteria. All these requirements are met by real time Polymerase Chain Reaction (or quantitative PCR; qPCR). Here, we compare existing methods of isolating DNA and their adaptation to a red wine matrix. Two different protocols for isolating DNA and three PCR mix compositions were tested to select the best method. The addition of insoluble polyvinylpolypyrrolidone (PVPP) at 1% (v/v) during DNA extraction using a protocol succeeded in eliminating PCR inhibitors from red wine. We developed a bacterial internal control which was efficient in avoiding false negative results due to decreases in the efficiency of DNA isolation and/or amplification. The specificity, linearity, repeatability, and reproducibility of the method were evaluated. A standard curve was established for the enumeration of AAB inoculated into red wines. The limit of quantification in red wine was 3.7 log AAB/mL and about 2.8 log AAB/mL when the volume of the samples was increased from 1 to 10 mL. Thus, the DNA extraction method developed in this paper allows sensitive and reliable AAB quantification without underestimation thanks to the presence of an internal control. Moreover, monitoring of both the AAB population and the amount of acetic acid in ethanol medium and red wine highlighted that a minimum about 6.0 log cells/mL of AAB is needed to significantly increase the production of acetic acid leading to spoilage. PMID:27313572

  9. Extraction mechanism of Sc(III) and separation from Th(IV), Fe(III) and Lu(III) with bis(2,4,4-trimethylpentyl)phosphinic acid in n-hexane from sulphuric acid solutions

    SciTech Connect

    Wang, Chun; Li, Deqian )

    1994-06-01

    The extraction equilibrium data of sulphuric acid and scandium(III) with bis(2,4,4-trimethylpentyl)phosphinic acid (H[BTMPP]) from sulphuric acid solutions have been obtained. There are two extraction mechanisms of scandium(III) with H[BTMPP] at different ranges of aqueous acidity. The extraction of Sc(III) proceeds according to a cation exchange reaction at lower aqueous acidity and to a solvating reaction at higher acidity. The practicability of separating Sc(III) from Th(IV),Fe(III) and Lu(III) by controlling the aqueous acidity was discussed. The effect of temperature on extraction of Sc(III) was observed and thermodynamic functions were calculated. IR and NMR spectra were made for organic extracts. The extraction characteristics of H[BTMPP] and di(2-ethylhexyl)phosphinic acid (H[DEHP]) were contrasted to obtain more information about extraction with H[BTMPP]. 12 refs., 6 figs., 1 tab.

  10. Assessment of bioavailable organic phosphorus in tropical forest soils by organic acid extraction and phosphatase hydrolysis.

    PubMed

    Darch, Tegan; Blackwell, Martin S A; Chadwick, David; Haygarth, Philip M; Hawkins, Jane M B; Turner, Benjamin L

    2016-12-15

    Soil organic phosphorus contributes to the nutrition of tropical trees, but is not accounted for in standard soil phosphorus tests. Plants and microbes can release organic anions to solubilize organic phosphorus from soil surfaces, and synthesize phosphatases to release inorganic phosphate from the solubilized compounds. We developed a procedure to estimate bioavailable organic phosphorus in tropical forest soils by simulating the secretion processes of organic acids and phosphatases. Five lowland tropical forest soils with contrasting properties (pH 4.4-6.1, total P 86-429 mg P kg(- 1)) were extracted with 2 mM citric acid (i.e., 10 μmol g(- 1), approximating rhizosphere concentrations) adjusted to soil pH in a 4:1 solution to soil ratio for 1 h. Three phosphatase enzymes were then added to the soil extract to determine the forms of hydrolysable organic phosphorus. Total phosphorus extracted by the procedure ranged between 3.22 and 8.06 mg P kg(- 1) (mean 5.55 ± 0.42 mg P kg(- 1)), of which on average three quarters was unreactive phosphorus (i.e., organic phosphorus plus inorganic polyphosphate). Of the enzyme-hydrolysable unreactive phosphorus, 28% was simple phosphomonoesters hydrolyzed by phosphomonoesterase from bovine intestinal mucosa, a further 18% was phosphodiesters hydrolyzed by a combination of nuclease from Penicillium citrinum and phosphomonoesterase, and the remaining 51% was hydrolyzed by a broad-spectrum phytase from wheat. We conclude that soil organic phosphorus can be solubilized and hydrolyzed by a combination of organic acids and phosphatase enzymes in lowland tropical forest soils, indicating that this pathway could make a significant contribution to biological phosphorus acquisition in tropical forests. Furthermore, we have developed a method that can be used to assess the bioavailability of this soil organic phosphorus.

  11. Advancing forensic RNA typing: On non-target secretions, a nasal mucosa marker, a differential co-extraction protocol and the sensitivity of DNA and RNA profiling.

    PubMed

    van den Berge, Margreet; Bhoelai, Bryan; Harteveld, Joyce; Matai, Anuska; Sijen, Titia

    2016-01-01

    The forensic identification of human body fluids and tissues by means of messenger RNA (mRNA) profiling is a long studied methodology that is increasingly applied to casework samples. Previously, we have described an mRNA multiplex system that targets blood, saliva, semen, menstrual secretion, vaginal mucosa and skin (Lindenbergh et al. and van den Berge et al.). In this study we consider various topics to improve this mRNA profiling system or its use and adapt the method accordingly. Bodily secretions that may be encountered at a crime scene whilst not targeted by the multiplex-id est nasal mucosa, sweat, tears, faeces and urine-were examined for false positive signals. The results prompted us to identify a nasal mucosa marker that allows the discrimination of nasal mucosa from saliva or vaginal mucosa and nosebleed blood from peripheral blood. An updated version of the multiplex was prepared to which the nasal mucosa marker was added and in which markers for semen, vaginal mucosa and blood were replaced. Lactobacillus markers were regarded unsuitable as replacement for vaginal mucosa mRNA markers because of background signals on penile swabs that appeared devoid of female DNA. Furthermore, we provide approaches to deal with highly unbalanced mixtures. First, a differential extraction protocol was incorporated into a co-extraction protocol to allow DNA and RNA analysis of separated non-sperm and sperm fractions. In a second approach, besides the standard multiplex, a customized multiplex is used which excludes markers for prevailing cell types. This allows the use of lower cDNA inputs for the prevailing cell types and higher inputs for cell types that appear masked. Additionally, we assessed the relation between the percentage of alleles or markers detected in DNA or RNA profiles when decreasing sample amounts are analysed. While blood, saliva, semen and menstrual secretion show the trend that DNA profiling is more sensitive than RNA profiling, the reverse is seen

  12. Binding characteristics of Cu(2+) to natural humic acid fractions sequentially extracted from the lake sediments.

    PubMed

    He, En; Lü, Changwei; He, Jiang; Zhao, Boyi; Wang, Jinghua; Zhang, Ruiqing; Ding, Tao

    2016-11-01

    Humic acids (HAs) determine the distribution, toxicity, bioavailability, and ultimate fate of heavy metals in the environment. In this work, ten HA fractions (F1-F10) were used as adsorbent, which were sequentially extracted from natural sediments of Lake Wuliangsuhai, to investigate the binding characteristics of Cu(2+) to HA. On the basis of the characterization results, differences were found between the ten extracted HA fractions responding to their elemental compositions and acidic functional groups. The characterization results reveal that the responses of ten extracted HA fractions to their elemental compositions and acidic functional groups were different. The O/C and (O + N)/C ratio of F1-F8 approximately ranged from 0.66 to 0.53 and from 0.72 to 0.61, respectively; the measured results showed that the contents of phenolic groups and carboxyl groups decreased from 4.46 to 2.60 mmol/g and 1.60 to 0.58 mmol/g, respectively. The binding characteristics of Cu(2+) to the ten HA fractions were well modeled by the bi-Langmuir model; the binding behavior of Cu(2+) to all the ten HA fractions were strongly impacted by pH and ionic strength. The FTIR and SEM-EDX image of HA fractions (pre- and post-adsorption) revealed that carboxyl and phenolic groups were responsible for the Cu(2+) sorption on the ten sequentially extracted HA fractions process, which is the same with the analysis of the ligand binding and bi-Langmuir models Accordingly, the adsorption capacity of the former HA fractions on Cu(2+) were higher than the latter ones, which may be attributed to the difference of carboxyl and phenolic group contents between the former and latter extracted HA fractions. Additionally, the functional groups with N and S should not be neglected. This work is hopeful to understand the environmental effect of humic substances, environmental geochemical behavior, and bioavailability of heavy metals in lakes.

  13. Rapid determination of acid herbicides in soil by liquid chromatography with tandem mass spectrometric detection based on dispersive solid phase extraction.

    PubMed

    Kaczyński, Piotr; Łozowicka, Bożena; Jankowska, Magdalena; Hrynko, Izabela

    2016-05-15

    This study determined twenty six the highly sensitive phenoxy, pyridines, aliphatic and aromatic acid compounds in soil with a liquid chromatography tandem mass spectrometry. The samples were prepared by modified quick, easy, cheap, effective, rugged and safe (QuEChERS) analytical procedure in solid samples. Herbicides extraction effectiveness was evaluated at three different spiking levels (0.01, 0.1 and 1.0 mg kg(-1)). Fourteen different dispersive solid-phase extraction (d-SPE) sorbents in clean-up step were tested. The QuEChERS protocol with acidic alumina provided the highest number of pesticides with recoveries in the 70-120% range. The soil matrix effect was evaluated and for the majority of compounds were not significant, showing suppression or enhancement (±81-123%). The precision calculated as relative standard deviation (RSD) was below 22%. The linear relation was observed in the range 0.01-2.0 mg kg(-1) and the correlation coefficient R>0.999. The expanded measurement uncertainty was estimated as being on average, and was between 9% and 33%. The validated method was employed in the analysis of 309 real soil samples.

  14. Extraction and separation of Pd(II), Pt(IV), Fe(III), Zn(II), Cu(II) and Ag(I) from hydrochloric acid solutions with selected cyanamides as novel extractants.

    PubMed

    Mowafy, E A; Aly, H F

    2007-10-22

    Three structurally related novel extractants namely: N,N-dihexylcyanamide (DHCY), N,N-di(2-ethylhexyl)cyanamide (DEHCY) and N,N-di-octylcyanamide (DOCY) were synthesized in our laboratory and characterized by different techniques. The general method for synthesizing these extractants was based on the reaction of relevant secondary amines with cyanogen bromide in presence of sodium acetate anhydride. Their extracting ability in toluene as a diluent for Pd(II), Pt(IV), Fe(III), Zn(II), Cu(II) and Ag(I) from hydrochloric acid media has been studied. The extraction of hydrochloric acid was studied also. Pd(II) was strongly extracted by these extractants at low hydrochloric acid concentrations and the extraction decreased with increasing hydrochloric acid concentration while the reverse was obtained in the extraction of Pt(IV), Fe(III) and Zn(II). Under similar extraction conditions Cu(II) and Ag(I) were found poorly extracted. Hydrochloric acid was extracted only in its high concentration region. A systematic investigation has been carried out on the extraction of Pd(II) using two of the synthesized extractants. Pd(II) was extracted as a solvated complex with the composition, metal:chloride ion:extractant=1:2:2. The extracted species were studied also using IR spectra.

  15. Extracorporeal worm extraction of Diphyllobothrium nihonkaiense with amidotrizoic acid in a child.

    PubMed

    Shin, Hye Kyung; Roh, Joo-Hyung; Oh, Jae-Won; Ryu, Jae-Sook; Goo, Youn-Kyoung; Chung, Dong-Il; Kim, Yong Joo

    2014-12-01

    Infection cases of diphyllobothriid tapeworms are not much in the below teen-age group. We report a case of Diphyllobothrium nihonkaiense infection in a 13-year-old boy. He presented with severe fatigue, occasional abdominal pain at night time. He also had several episodes of tapeworm segment discharge in his stools. By his past history, he had frequently eaten raw fish including salmon and trout with his families. Numerous eggs of diphyllobothriid tapeworm were detected in the fecal examination. We introduced amidotrizoic acid as a cathartic agent through nasogastroduodenal tube and let nearly whole length (4.75 m) of D. nihonkaiense be excreted through his anus. After a single dose of praziquantel, the child's stool showed no further eggs, and his symptoms disappeared. The evacuated worm was identified as D. nihonkaiense by mitochondrial cox1 gene analysis. Here we report a successful extracorporeal worm extraction from an infection case of D. nihonkaiense by the injection of amidotrizoic acid.

  16. Extracorporeal Worm Extraction of Diphyllobothrium nihonkaiense with Amidotrizoic Acid in a Child

    PubMed Central

    Shin, Hye Kyung; Roh, Joo-Hyung; Oh, Jae-Won; Ryu, Jae-Sook; Goo, Youn-Kyoung; Chung, Dong-Il

    2014-01-01

    Infection cases of diphyllobothriid tapeworms are not much in the below teen-age group. We report a case of Diphyllobothrium nihonkaiense infection in a 13-year-old boy. He presented with severe fatigue, occasional abdominal pain at night time. He also had several episodes of tapeworm segment discharge in his stools. By his past history, he had frequently eaten raw fish including salmon and trout with his families. Numerous eggs of diphyllobothriid tapeworm were detected in the fecal examination. We introduced amidotrizoic acid as a cathartic agent through nasogastroduodenal tube and let nearly whole length (4.75 m) of D. nihonkaiense be excreted through his anus. After a single dose of praziquantel, the child's stool showed no further eggs, and his symptoms disappeared. The evacuated worm was identified as D. nihonkaiense by mitochondrial cox1 gene analysis. Here we report a successful extracorporeal worm extraction from an infection case of D. nihonkaiense by the injection of amidotrizoic acid. PMID:25548421

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

    PubMed

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

    2012-12-04

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

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

    PubMed

    Reddersen, K; Heberer, Th

    2003-09-05

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

  19. Development of a solid-phase extraction method for simultaneous extraction of adipic acid, succinic acid and 1,4-butanediol formed during hydrolysis of poly(butylene adipate) and poly(butylene succinate).

    PubMed

    Lindström, Annika; Albertsson, Ann-Christine; Hakkarainen, Minna

    2004-01-02

    A solid-phase extraction (SPE) method was developed for the simultaneous extraction of dicarboxylic acids and diols formed during hydrolysis of poly(butylene succinate), PBS, and poly(butylene adipate), PBA. Four commercial non-polar SPE columns, three silica based: C8, C18, C18 (EC), and one resin based: ENV+, were tested for the extraction of succinic acid, adipic acid and 1,4-butanediol, the expected final hydrolysis products of PBS and PBA. ENV+ resin was chosen as a solid-phase, because it displayed the best extraction efficiency for 1,4-butanediol and succinic acid. Linear range for the extracted analytes was 1-500 ng/microl for adipic acid and 2-500 ng/microl for 1,4-butanediol and succinic acid. Detection and quantification limits for the analytes were between 1-2 and 2-7 ng/microl, respectively, and relative standard deviations were between 3 and 7%. Good repeatability and low detection limits made the developed SPE method and subsequent gas chromatography-mass spectrometry (GC-MS) analysis a sensitive tool for identification and quantification of hydrolysis products at early stages of degradation.

  20. Application of ionic liquids based microwave-assisted simultaneous extraction of carnosic acid, rosmarinic acid and essential oil from Rosmarinus officinalis.

    PubMed

    Liu, Tingting; Sui, Xiaoyu; Zhang, Rongrui; Yang, Lei; Zu, Yuangang; Zhang, Lin; Zhang, Ying; Zhang, Zhonghua

    2011-11-25

    An ionic liquid based microwave-assisted simultaneous extraction and distillation (ILMSED) method has been developed for the effective extraction of carnosic acid (CA), rosmarinic acid (RA) and essential oil (EO) from Rosmarinus officinalis. A series of 1-alkyl-3-methylimidazolium ionic liquids differing in composition of anion and cation were evaluated for extraction yield in this work. The results obtained indicated that the anions and cations of ionic liquids had influences on the extraction of CA and RA, 1.0M 1-octyl-3-methylimidazolium bromide ([C8mim]Br) solution was selected as solvent. In addition, the ILMSED procedures for the three target ingredients were optimized and compared with other conventional extraction techniques. ILMSED gave the best result due to the highest extraction yield within the shortest extraction time for CA and RA. The novel process developed offered advantages in term of yield and selectivity of EO and shorter isolation time (20 min in comparison of 4h of hydrodistillation), and provides a more valuable EO (with high amount of oxygenated compounds). The microstructures and chemical structures of rosemary samples before and after extraction were also investigated. Moreover, the proposed method was validated by the stability, repeatability and recovery experiments. The results indicated that the developed ILMSED method provided a good alternative for the both extraction of non-volatile compounds (CA and RA) and EO from rosemary as well as other herbs.

  1. Stability of Rosmarinic Acid in Aqueous Extracts from Different Lamiaceae Species after in vitro Digestion with Human Gastrointestinal Enzymes

    PubMed Central

    Zorić, Zoran; Markić, Joško; Pedisić, Sandra; Bučević-Popović, Viljemka; Generalić-Mekinić, Ivana; Grebenar, Katarina

    2016-01-01

    Summary The present study compares the gastrointestinal stability of rosmarinic acid in aqueous extracts of thyme, winter savory and lemon balm with the stability of pure rosmarinic acid. The stability of rosmarinic acid was detected after two-phase in vitro digestion process (gastric and duodenal) with human gastrointestinal enzymes. The concentration of rosmarinic acid in undigested and digested samples was detected using HPLC-DAD. Results showed that gastrointestinal stability of pure rosmarinic acid was significantly higher than that of rosmarinic acid from plant extracts after both gastric and intestinal phases of digestion. Among plant extracts, rosmarinic acid was the most stable in lemon balm after gastric (14.10%) and intestinal digestion phases (6.5%). The temperature (37 °C) and slightly alkaline medium (pH=7.5) did not affect the stability of rosmarinic acid, while acid medium (pH=2.5) significantly decreased its stability (≥50%). In addition, the stability rate of rosmarinic acid is influenced by the concentration of human gastrointestinal juices. PMID:27904398

  2. Stability of Rosmarinic Acid in Aqueous Extracts from Different Lamiaceae Species after in vitro Digestion with Human Gastrointestinal Enzymes.

    PubMed

    Zorić, Zoran; Markić, Joško; Pedisić, Sandra; Bučević-Popović, Viljemka; Generalić-Mekinić, Ivana; Grebenar, Katarina; Kulišić-Bilušić, Tea

    2016-03-01

    The present study compares the gastrointestinal stability of rosmarinic acid in aqueous extracts of thyme, winter savory and lemon balm with the stability of pure rosmarinic acid. The stability of rosmarinic acid was detected after two-phase in vitro digestion process (gastric and duodenal) with human gastrointestinal enzymes. The concentration of rosmarinic acid in undigested and digested samples was detected using HPLC-DAD. Results showed that gastrointestinal stability of pure rosmarinic acid was significantly higher than that of rosmarinic acid from plant extracts after both gastric and intestinal phases of digestion. Among plant extracts, rosmarinic acid was the most stable in lemon balm after gastric (14.10%) and intestinal digestion phases (6.5%). The temperature (37 °C) and slightly alkaline medium (pH=7.5) did not affect the stability of rosmarinic acid, while acid medium (pH=2.5) significantly decreased its stability (≥50%). In addition, the stability rate of rosmarinic acid is influenced by the concentration of human gastrointestinal juices.

  3. EFFECT OF PERILLA FRUTESCENS EXTRACTS AND ROSMARINIC ACID ON RAT HEART MITOCHONDRIAL FUNCTIONS.

    PubMed

    Raudone, Lina; Burdulis, Deividas; Raudonis, Raimondas; Janulis, Valdimaras; Jankauskiene, Laima; Viskelis, Pranas; Trumbeckaite, Sonata

    2016-01-01

    Perilla frutescens L. due to its aromatic, antibacterial, anti-inflammatory and antioxidant traits has been traditionally used as medicinal plant in Eastern Asia. Alterations of mitochondria are interconnected with many chronic diseases. Bioactives of herbal extracts can modulate mitochondrial effects and be beneficial in prevention of mitochondrial related chronic diseases. Direct effects of the red-leaf form P. frutescens extract (PFE) and the green-leaf form P. frutescens var. crispa f. viridis extract (PCE) were evaluated investigating activities on the oxidative phosphorylation and antioxidant activity in the rat heart mitochondria in vitro. HPLC-MS analysis was applied for the identification of phenolic compounds. Cell with a Clark-type oxygen electrode was used for mitochondrial respiration measurement. The generation of reactive oxygen species was estimated in isolated rat heart mitochondria and determined fluorimetrically. State 3 respiration rate was not affected by lower concentrations, however, it was inhibited at higher concentrations by 22-70% for PFE and by 45-55% for PCE. PFE containing anthocyanins induced the concentration-dependent stimulation (by 23-76%) of the State 4 respiration rate after addition of cytochrome c due to reducing properties. Significant reduction of H₂O₂ pro- duction was observed with investigated concentrations of rosmarinic acid and both perilla extracts. Our results demonstrate that the effect of PFE and PCE extracts on rat heart mitochondria depend on the qualitative characteristics of complex of biologically active compounds. Selective effects on mitochondrial function could enable the regulation of apoptosis or another mechanisms occurring in cells.

  4. New method for the rapid extraction of natural products: efficient isolation of shikimic acid from star anise.

    PubMed

    Just, Jeremy; Deans, Bianca J; Olivier, Wesley J; Paull, Brett; Bissember, Alex C; Smith, Jason A

    2015-05-15

    A new, practical, rapid, and high-yielding process for the pressurized hot water extraction (PHWE) of multigram quantities of shikimic acid from star anise (Illicium verum) using an unmodified household espresso machine has been developed. This operationally simple and inexpensive method enables the efficient and straightforward isolation of shikimic acid and the facile preparation of a range of its synthetic derivatives.

  5. Effects of radiation, acid, and base on the extractant dihexyl-(diethylcarbamoyl)methyl) phosphonate

    SciTech Connect

    Bahner, C.T.; Shoun, R.R.; McDowell, W.J.

    1981-11-01

    The effects of exposure to gamma radiation (/sup 60/Co) and of contact with acidic and basic aqueous solutions on dihexyl((diethylcarbamoyl)methyl)phosphonate (DHDECMP) were studied. Gamma radiation decomposes DHDECMP into a variety of products. The most troublesome of those are the acidic compounds that cause problems in stripping the actinides and lanthanides from the extractant at low acid concentrations. The rate of degradation of DHDECMP by radiation is about the same or only slightly higher than that of tri-n-butyl phosphate (TBP). It is relatively easy to remove the radiation-produced impurities by equilibration (scrubbing) with sodium carbonate or sodium hydroxide or by column chromatographic methods. The hydrolysis of DHDECMP in contact with aqueous solutions containing less than 3 M HNO/sub 3/ is not more severe than that of TBP under the same conditions but is significant above that acid concentration. Hydrolysis of DHDECMP in contact with aqueous sodium hydroxide solution does occur, but it should not pose an important problem with the short contact times such as those anticipated for the removal of the radiation-induced degradation products by caustic scrubbing. Results of various chromatographic tests to characterize the degradation products of DHDECMP are also given.

  6. Extraction of vanadium from vanadium slag by high pressure oxidative acid leaching

    NASA Astrophysics Data System (ADS)

    Zhang, Guo-quan; Zhang, Ting-an; Lü, Guo-zhi; Zhang, Ying; Liu, Yan; Liu, Zhuo-lin

    2015-01-01

    To extract vanadium in an environment friendly manner, this study focuses on the process of leaching vanadium from vanadium slag by high pressure oxidative acid leaching. Characterizations of the raw slag, mineralogy transformation, and the form of leach residues were made by X-ray diffraction, scanning electron microscopy, and energy dispersive X-ray spectroscopy. The result shows that the vanadium slag is composed of major phases of fayalite, titanomagnetite, and spinel. During the high pressure oxidative acid leaching process, the fayalite and spinel phases are gradually decomposed by sulfuric acid, causing the release of vanadium and iron in the solution. Meanwhile, unreacted silicon and titanium are enriched in the leach residues. With the initial concentration of sulfuric acid at 250 g·L-1, a leaching temperature of 140°C, a leaching time of 50 min, a liquid-solid ratio of 10:1 mL·g-1, and oxygen pressure at 0.2 MPa, the leaching rate of vanadium reaches 97.69%.

  7. Effect of acidity on the energy level of curcumin dye extracted from Curcuma longa L.

    NASA Astrophysics Data System (ADS)

    Agustia, Yuda Virgantara; Suyitno, Arifin, Zainal; Sutanto, Bayu

    2016-03-01

    The purpose of this research is to investigate the effect of acidity on the energy level of curcumin dye. The natural dye, curcumin, was synthesized from Curcuma longa L. using a simple extraction technique. The purification of curcumin dye was conducted in a column of chromatography and its characteristics were studied. Next, the purified curcumin dye was added by benzoic acids until various acidities of 3.0, 3.5, 4.0, 4.5, and 5.0. The absorbance spectra and the functionality groups found in the dyes were detected by ultraviolet-visible spectroscopy and Fourier-transform infrared spectroscopy, respectively. Meanwhile, the energy level of the dyes, EHOMO and ELUMO was measured by cyclic voltammetry. The best energy level of curcumin dye was achieved at pH 3.5 where Ered = -0.37V, ELUMO = -4.28 eV, Eox = 1.15V, EHOMO = -5.83 eV, and Eband gap = 1.55 eV. Therefore, the purified curcumin dye added by benzoic acid was promising for sensitizing the dye-sensitized solar cells.

  8. Production of (R)-3-hydroxybutyric acid by Burkholderia cepacia from wood extract hydrolysates

    PubMed Central

    2014-01-01

    (R)-hydroxyalkanoic acids (R-HAs) are valuable building blocks for the synthesis of fine chemicals and biopolymers because of the chiral center and the two active functional groups. Hydroxyalkanoic acids fermentation can revolutionize the polyhydroxyalkanoic acids (PHA) production by increasing efficiency and enhancing product utility. Modifying the fermentation conditions that promotes the in vivo depolymerization and secretion to fermentation broth in wild type bacteria is a novel and promising approach to produce R-HAs. Wood extract hydrolysate (WEH) was found to be a suitable substrate for R-3-hydroxybutyric acid (R-3-HB) production by Burkholderia cepacia. Using Paulownia elongate WEH as a feedstock, the R-3-HB concentration in fermentation broth reached as high as 14.2 g/L after 3 days of batch fermentation and the highest concentration of 16.8 g/L was obtained at day 9. Further investigation indicated that the composition of culture medium contributed to the enhanced R-3-HB production. PMID:24949263

  9. Phytochemical Profiling of Flavonoids, Phenolic Acids, Terpenoids, and Volatile Fraction of a Rosemary (Rosmarinus officinalis L.) Extract.

    PubMed

    Mena, Pedro; Cirlini, Martina; Tassotti, Michele; Herrlinger, Kelli A; Dall'Asta, Chiara; Del Rio, Daniele

    2016-11-19

    This paper presents a comprehensive analysis of the phytochemical profile of a proprietary rosemary (Rosmarinus officinalis L.) extract rich in carnosic acid. A characterization of the (poly)phenolic and volatile fractions of the extract was carried out using mass spectrometric techniques. The (poly)phenolic composition was assessed by ultra-high performance liquid chromatography-electrospray ionization-mass spectrometry (UHPLC-ESI-MS(n)) and a total of 57 compounds were tentatively identified and quantified, 14 of these being detected in rosemary extract for the first time. The rosemary extract contained 24 flavonoids (mainly flavones, although flavonols and flavanones were also detected), 5 phenolic acids, 24 diterpenoids (carnosic acid, carnosol, and rosmanol derivatives), 1 triterpenoid (betulinic acid), and 3 lignans (medioresinol derivatives). Carnosic acid was the predominant phenolic compound. The volatile profile of the rosemary extract was evaluated by head space solid-phase microextraction (HS-SPME) linked to gas chromatography-mass spectrometry (GC-MS). Sixty-three volatile molecules (mainly terpenes, alcohols, esters, aldehydes, and ketones) were identified. This characterization extends the current knowledge on the phytochemistry of Rosmarinus officinalis and is, to our knowledge, the broadest profiling of its secondary metabolites to date. It can assist in the authentication of rosemary extracts or rosemary-containing products or in testing its bioactivity. Moreover, this methodological approach could be applied to the study of other plant-based food ingredients.

  10. The impact of extraction with a chelating agent under acidic conditions on the cell wall polymers of mango peel.

    PubMed

    Jamsazzadeh Kermani, Zahra; Shpigelman, Avi; Kyomugasho, Clare; Van Buggenhout, Sandy; Ramezani, Mohsen; Van Loey, Ann M; Hendrickx, Marc E

    2014-10-15

    The objective of this research was to evaluate whether mango peel is a potential source of functional cell wall polymers. To reach this objective, the native pectin polymers (NPP) extracted as alcohol insoluble residue from mango peel, were characterised in terms of uronic acid content, degree of methoxylation, neutral sugar content, and molar mass and compared to citric acid (pH 2.5, 2h at 80°C) extracted polymers, mimicking industrial pectin extraction conditions. Water-solubilised NPP were highly methoxylated having two populations with a Mw of 904 and 83kDa and a degree of methoxylation of 66%. Citric acid extraction with a yield higher than H2SO4 extraction resulted in a very branched pectin with an extremely high DM (83%) and a high molar mass. Comparing the Fourier Transform Infra-Red spectroscopy of extracted and native WSF showed that citric acid remained partially associated to the extracted pectin due to its chelating properties.

  11. DIAMIDE DERIVATIVES OF DIPICOLINIC ACID AS ACTINIDE AND LANTHANIDE EXTRACTANTS IN A VARIATION OF THE UNEX PROCESS

    SciTech Connect

    D. R. Peterman; R. S. Herbst; J. D. Law; R. D. Tillotson; T. G. Garn; T. A. Todd; V. N. Romanovskiy; V. A. Babain; M. Yu. Alyapyshev; I. V. Smirnov

    2007-09-01

    The Universal Extraction (UNEX) process has been developed for simultaneous extraction of cesium, strontium, and actinides from acidic solutions. This process utilizes an extractant consisting of 0.08 M chlorinated cobalt dicarbollide (HCCD), 0.007-0.02 M polyethylene glycol (PEG-400), and 0.02 M diphenyl-N,N-di-n-butylcarbamoylmethylphosphine oxide (Ph2CMPO) in the diluent trifluoromethylphenyl sulfone (CF3C6H5SO2, designated FS-13) and provides simultaneous extraction of Cs, Sr, actinides, and lanthanides from HNO3 solutions. The UNEX process is of limited utility for processing acidic solutions containing large quantities of lanthanides and/or actinides, such as dissolved spent nuclear fuel solutions. These constraints are primarily attributed to the limited concentrations of CMPO (a maximum of ~0.02 M) in the organic phase and limited solubility of the CMPO-metal complexes. As a result, alternative actinide and lanthanide extractants are being investigated for use with HCCD as an improvement for waste processing and for applications where higher concentrations of the metals are present. Our preliminary results indicate that diamide derivatives of dipicolinic acid may function as efficient actinide and lanthanide extractants. The results to be presented indicate that, of the numerous diamides studied to date, the tetrabutyldiamide of dipicolinic acid, TBDPA, shows the most promise as an alternative actinide/lanthanide extractant in the UNEX process.

  12. Ultrasound-assisted extraction of pectins from grape pomace using citric acid: a response surface methodology approach.

    PubMed

    Minjares-Fuentes, R; Femenia, A; Garau, M C; Meza-Velázquez, J A; Simal, S; Rosselló, C

    2014-06-15

    An ultrasound-assisted procedure for the extraction of pectins from grape pomace with citric acid as the extracting agent was established. A Box-Behnken design (BBD) was employed to optimize the extraction temperature (X1: 35-75°C), extraction time (X2: 20-60 min) and pH (X3: 1.0-2.0) to obtain a high yield of pectins with high average molecular weight (MW) and degree of esterification (DE) from grape pomace. Analysis of variance showed that the contribution of a quadratic model was significant for the pectin extraction yield and for pectin MW whereas the DE of pectins was more influenced by a linear model. An optimization study using response surface methodology was performed and 3D response surfaces were plotted from the mathematical model. According to the RSM model, the highest pectin yield (∼32.3%) can be achieved when the UAE process is carried out at 75°C for 60 min using a citric acid solution of pH 2.0. These pectic polysaccharides, composed mainly by galacturonic acid units (<97% of total sugars), have an average MW of 163.9 kDa and a DE of 55.2%. Close agreement between experimental and predicted values was found. These results suggest that ultrasound-assisted extraction could be a good option for the extraction of functional pectins with citric acid from grape pomace at industrial level.

  13. Survival of Unstressed and Acid-, Cold-, and Starvation-Stress-Adapted Listeria monocytogenes in Ham Extract with Hops Beta Acids and Consumer Acceptability of HBA on Ready-to-Eat Ham

    PubMed Central

    Wang, Li; Shen, Cangliang

    2015-01-01

    The efficacy of hops beta acids (HBA) against unstressed and stress-adapted Listeria monocytogenes in ham extract and the consumers' acceptability of HBA on ready-to-eat (RTE) hams were investigated. Unstressed or acid-, cold-, or starvation-stress-adapted L. monocytogenes was inoculated (1.3–1.5 log CFU/mL) into 10% ham extract, without (control) or with HBA (4.44 or 10.0 µg/mL). Survival/growth of the pathogen during storage (7.2°C, 26 days) was moni