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Sample records for single-step liquid chromatography

  1. Online polar two phase countercurrent chromatography×high performance liquid chromatography for preparative isolation of polar polyphenols from tea extract in a single step.

    PubMed

    Chen, Wei-Bin; Li, Shu-Qi; Chen, Long-Jiang; Fang, Mei-Juan; Chen, Quan-Cheng; Wu, Zhen; Wu, Yun-Long; Qiu, Ying-Kun

    2015-08-01

    Herein, we report an on-line two-dimensional system constructed by counter-current chromatography (CCC) coupling with preparative high-performance liquid chromatography (prep-HPLC) for the separation and purification of polar natural products. The CCC was used as the first dimensional isolation column, where an environmental friendly polar two-phase solvent system of isopropanol and 16% sodium chloride aqueous solution (1:1.2, v/v) was introduced for low toxicity and favorable resolution. In addition, by applying the stop-and-go flow technique, effluents pre-fractionated by CCC was further purified by a preparative column packed with octadecyl silane (ODS) as the second dimension. The interface between the two dimensions was comprised of a 6-port switching valve and an electronically controlled 2-position 10-port switching valve connected with two equivalent holding columns. To be highlighted here, this rationally designed interface for the purpose of smooth desalination, absorption and desorption, successfully solved the solvent compatibility problem between the two dimensional separation systems. The present integrated system was successfully applied in a one-step preparative separation and identification of 10 pure compounds from the water extracts of Tieguanyin tea (Chinese oolong tea). In short, all the results demonstrated that the on-line 2D CCC×LC method is an efficient and green approach for harvesting polar targets in a single step, which showed great promise in drug discovery.

  2. A single-step solid phase extraction for the simultaneous determination of 8 mycotoxins in fruits by ultra-high performance liquid chromatography tandem mass spectrometry.

    PubMed

    Wang, Meng; Jiang, Nan; Xian, Hong; Wei, Dizhe; Shi, Lei; Feng, Xiaoyuan

    2016-01-15

    A simple and rapid extraction procedure for the simultaneous determination of eight mycotoxins (Alternaria toxins, ochratoxin A, patulin, citrinin) in a variety of fruit matrices has been developed using ultra high performance liquid chromatography coupled to tandem mass spectrometry. The procedure involves a one-step cleanup using homemade solid phase extraction (SPE) cartridges. By comparative evaluation among six various adsorbents (C18, PSA, HLB, MCX, Silica, NH2), the combination of MCX and NH2 was found to provide the most effective cleanup, removing the greatest number of matrix interferences and also allowing the quantification of all analyzed mycotoxins in fruits. The optimized extraction conditions including acidified aqueous acetonitrile and an additional salt-out step using NaCl were employed before SPE cleanup. Method validation was performed by analyzing samples spiked at three levels (LOQ, 2 LOQ and 10 LOQ). Four fruits including apple, sweet cherry, tomato and orange fruits were selected, and accuracy (recovery%), precision (RSD%), limits of quantification (LOQ), linearity and matrix effect were evaluated during validation. Matrix-matched linearity with correlation coefficients ≥ 0.9921 was established in the range of 5-200 ng mL(-1) for patulin and 1-200 ng mL(-1) for other mycotoxins, respectively. Recoveries between 74.2% and 102.4% and relative standard deviations lower than 4.7% were obtained for all tested fruits. The matrix effect observed was low (≤ ± 17%) in all three fruit matrixes with the exception of orange, for which strong ion suppression was observed for alternariol (25.3%), ochratoxin A (31.6%) and citrinin (40.3%). Therefore, matrix-matched calibration was used for a correct quantification in order to compensate for matrix effect. The limits of quantification (LOQ), ranging from 1 to 5 μg kg(-1) depending on mycotoxins type, were always lower than maximum permitted levels for every regulated mycotoxin by the current European

  3. Single-step purification of native miraculin using immobilized metal-affinity chromatography.

    PubMed

    Duhita, Narendra; Hiwasa-Tanase, Kyoko; Yoshida, Shigeki; Ezura, Hiroshi

    2009-06-24

    Miraculin is a taste-modifying protein that can be isolated from miracle fruit ( Richadella dulcifica ), a shrub native to West Africa. It is able to turn a sour taste into a sweet taste. The commercial exploitation of this sweetness-modifying protein is underway, and a fast and efficient purification method to extract the protein is needed. We succeeded in purifying miraculin from miracle fruit in a single-step purification using immobilized metal-affinity chromatography (IMAC). The purified miraculin exhibited high purity (>95%) in reverse-phase high-performance liquid chromatography. We also demonstrated the necessity of its structure for binding to the nickel-IMAC column. PMID:19469504

  4. Combined Protein A and size exclusion high performance liquid chromatography for the single-step measurement of mAb, aggregates and host cell proteins.

    PubMed

    Gjoka, Xhorxhi; Schofield, Mark; Cvetkovic, Aleksandar; Gantier, Rene

    2014-12-01

    Quantification of monoclonal antibody (mAb) monomer, mAb aggregates, and host cell proteins (HCPs) is critical for the optimization of the mAb production process. The present work describes a single high throughput analytical tool capable of tracking the concentration of mAb, mAb aggregate and HCPs in a growing cell culture batch. By combining two analytical HPLC methods, Protein A affinity and size-exclusion chromatography (SEC), it is possible to detect a relative increase or decrease in the concentration of all three entities simultaneously. A comparison of the combined Protein A-SEC assay to SEC alone was performed, demonstrating that it can be useful tool for the quantification of mAb monomer along with trending data for mAb aggregate and HCP. Furthermore, the study shows that the Protein A-SEC method is at least as accurate as other commonly used analytical methods such as ELISA and Bradford.

  5. Single-step electrotransfer of reverse-stained proteins from sodium dodecyl sulfate-polyacrylamide gel onto reversed-phase minicartridge and subsequent desalting and elution with a conventional high-performance liquid chromatography gradient system for analysis.

    PubMed

    Fernandez-Patron, C; Madrazo, J; Hardy, E; Mendez, E; Frank, R; Castellanos-Serra, L

    1995-06-01

    Isolation of proteins from polyacrylamide electrophoresis gels by a novel combination of techniques is described. A given protein band from a reverse stained (imidazol-sodium dodecyl sulfate--zinc salts) gel can be directly electrotransferred onto a reversed-phase chromatographic support, packed in a self-made minicartridge (2 mm in thickness, 8 mm in internal diameter, made of inert polymeric materials). The minicartridge is then connected to a high-performance liquid chromatography system and the electrotransferred protein eluted by applying an acetonitrile gradient. Proteins elute in a small volume ( < 700 microL) of high-purity volatile solvents (water, trifluoroacetic acid, acetonitrile) and are free of contaminants (gel contaminants, salts, etc). Electrotransferred proteins were efficiently retained, e.g., up to 90% for radioiodinated alpha-lactalbumin, by the octadecyl matrix, and their recovery on elution from the minicartridge was in the range typical for this type of chromatographic support, e.g., 73% for alpha-lactalbumin. The technique was successfully applied to a variety of proteins in the molecular mass range 6-68 kDa, and with amounts between 50 and 2000 pmol. The good mechanical and chemical stability of the developed minicartridges, during electrotransfer and chromatography, allowed their repeated use. This new technique permitted a single-step separation of two proteins unresolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis due to their different elution from the reversed-phase support. The isolated proteins were amenable to analysis by N-terminal sequencing, enzymic digestion and mass spectrometry of their proteolytic fragments. Chromatographic elution of proteins from the reversed-phase mini-cartridge was apparently independent of the specific loading mode employed, i.e., loading by conventional loop injection or by electrotransfer. PMID:7498136

  6. Separation of Be and Al for AMS using single-step column chromatography

    NASA Astrophysics Data System (ADS)

    Binnie, Steven A.; Dunai, Tibor J.; Voronina, Elena; Goral, Tomasz; Heinze, Stefan; Dewald, Alfred

    2015-10-01

    With the aim of simplifying AMS target preparation procedures for TCN measurements we tested a new extraction chromatography approach which couples an anion exchange resin (WBEC) to a chelating resin (Beryllium resin) to separate Be and Al from dissolved quartz samples. Results show that WBEC-Beryllium resin stacks can be used to provide high purity Be and Al separations using a combination of hydrochloric/oxalic and nitric acid elutions. 10Be and 26Al concentrations from quartz samples prepared using more standard procedures are compared with results from replicate samples prepared using the coupled WBEC-Beryllium resin approach and show good agreement. The new column procedure is performed in a single step, reducing sample preparation times relative to more traditional methods of TCN target production.

  7. Single-step purification of chitosanases from Bacillus cereus using expanded bed chromatography.

    PubMed

    de Araújo, Nathália Kelly; Pagnoncelli, Maria Giovana Binder; Pimentel, Vanessa Carvalho; Xavier, Maria Luiza Oliveira; Padilha, Carlos Eduardo Araújo; de Macedo, Gorete Ribeiro; Dos Santos, Everaldo Silvino

    2016-01-01

    A chitosanase-producing strain was isolated and identified as Bacillus cereus C-01. The purification and characterization of two chitosanases were studied. The purification assay was accomplished by ion exchange expanded-bed chromatography. Experiments were carried out in the presence and in the absence of cells through different expansion degree to evaluate the process performance. The adsorption experiments demonstrated that the biomass does not affect substantially the adsorption capacity of the matrix. The enzyme bound to the resin with the same extent using clarified and unclarified broth (0.32 and 0.30 U/g adsorbent, respectively). The fraction recovered exhibited 31% of the yield with a 1.26-fold increase on the specific activity concerned to the initial broth. Two chitosanases from different elution steps were recovery. Chit A and Chit B were stable at 30-60°C, pH 5.5-8.0 and 5.5-7.5, respectively. The highest activity was found at 55°C, pH 5.5 to Chit A and 50°C, pH 6.5 to Chit B. The ions Cu(2+), Fe(2+) and Zn(2+) indicated inhibitory effect on chitosanases activities that were significantly activated by Mn(2+). The methodology applied in this study enables the partial purification of a stable chitosanase using a feedstock without any pre-treatment using a single-step purification.

  8. Single-step isolation of extracellular vesicles by size-exclusion chromatography

    PubMed Central

    Böing, Anita N.; van der Pol, Edwin; Grootemaat, Anita E.; Coumans, Frank A. W.; Sturk, Auguste; Nieuwland, Rienk

    2014-01-01

    Background Isolation of extracellular vesicles from plasma is a challenge due to the presence of proteins and lipoproteins. Isolation of vesicles using differential centrifugation or density-gradient ultracentrifugation results in co-isolation of contaminants such as protein aggregates and incomplete separation of vesicles from lipoproteins, respectively. Aim To develop a single-step protocol to isolate vesicles from human body fluids. Methods Platelet-free supernatant, derived from platelet concentrates, was loaded on a sepharose CL-2B column to perform size-exclusion chromatography (SEC; n=3). Fractions were collected and analysed by nanoparticle tracking analysis, resistive pulse sensing, flow cytometry and transmission electron microscopy. The concentrations of high-density lipoprotein cholesterol (HDL) and protein were measured in each fraction. Results Fractions 9–12 contained the highest concentrations of particles larger than 70 nm and platelet-derived vesicles (46%±6 and 61%±2 of totals present in all collected fractions, respectively), but less than 5% of HDL and less than 1% of protein (4.8%±1 and 0.65%±0.3, respectively). HDL was present mainly in fractions 18–20 (32%±2 of total), and protein in fractions 19–21 (36%±2 of total). Compared to the starting material, recovery of platelet-derived vesicles was 43%±23 in fractions 9–12, with an 8-fold and 70-fold enrichment compared to HDL and protein. Conclusions SEC efficiently isolates extracellular vesicles with a diameter larger than 70 nm from platelet-free supernatant of platelet concentrates. Application SEC will improve studies on the dimensional, structural and functional properties of extracellular vesicles. PMID:25279113

  9. Development of an aptamer-affinity chromatography for efficient single step purification of Concanavalin A from Canavalia ensiformis.

    PubMed

    Ahirwar, Rajesh; Nahar, Pradip

    2015-08-01

    Herein, an aptamer-based affinity chromatography method for rapid and single step purification of Concanavalin A is developed and validated. We have used a 41ntssDNA aptamer of Con A (Con A aptabody) as an affinity reagent in the developed aptamer-affinity chromatography. Stationary phase of the method consists of surface functionalized agarose beads carrying covalently immobilized Con A-aptabody. Affinity purification of Con A from jack bean (Canavalia ensiformis) seed using developed aptamer-affinity columns has resulted in ≥66% recovery with 90% purity and 336-fold purification of Con A. The developed aptamer-affinity chromatography has shown efficient scalability and consistent purification when analysed over 13mm, 20mm and 25mm diameter columns having a bed height of 60mm each. Also, the developed aptamer-agarose columns were found to be reusable with recovery decrease of 12.9% in seven sequential cycles of purification. Therefore, the developed aptamer-affinity chromatography provides a novel, efficient and single-step methodology for isolation and purification of Con A. PMID:26102634

  10. Development of a method to extract and purify target compounds from medicinal plants in a single step: online hyphenation of expanded bed adsorption chromatography and countercurrent chromatography

    PubMed Central

    Li, Yang; Wang, Nan; Zhang, Min; Ito, Yoichiro; Zhang, Hongyang; Wang, Yuerong; Guo, Xin; Hu, Ping

    2014-01-01

    Pure compounds extracted and purified from natural sources are crucial to lead discovery and drug screening. This study presents a novel two-dimensional hyphenation of expanded bed adsorption chromatography (EBAC) and high-speed countercurrent chromatography (HSCCC) for extraction and purification of target compounds from medicinal plants in a single step. The EBAC and HSCCC were hyphenated via a six-port injection valve as an interface. Fractionation of ingredients of Salvia miltiorrhiza and Rhizoma coptidis was performed on the hyphenated system to verify its efficacy. An amount each of 52.9 mg of salvianolic acid B and 2.1 mg of rosmarinic acid was obtained from Salvia miltiorrhiza by the two-dimensional system in a single step. The purities of the targets were over 96% of salvianolic acid B and 74% of rosmarinic acid. An amount each of 4.6 mg of coptisine and 4.1 mg of berberine was obtained from Rhizoma coptidis each with 98% and 82% purity, respectively. The processing time was nearly 50% that of the multi-step method. These results indicate that the present method is a rapid and green way to harvest targets from medicinal plants in a single step. PMID:24588208

  11. A single-step pesticide extraction and clean-up multi-residue analytical method by selective pressurized liquid extraction followed by on-line solid phase extraction and ultra-high-performance liquid chromatography-tandem mass spectrometry for complex matrices.

    PubMed

    Rodrigues, Elsa Teresa; Pardal, Miguel Ângelo; Salgueiro-González, Noelia; Muniategui-Lorenzo, Soledad; Alpendurada, Maria Fátima

    2016-06-24

    Pesticides, a group of compounds linked to human activity, may, when in toxic levels, have a profound effect on water quality, and hence result in adverse consequences to aquatic life and ultimately to human health. Analytical challenges arise when successfully trying to determine these levels in environmental complex matrices. Therefore, fast, simple, sensitive and selective analytical methodologies for multi-residue determination of pesticides (atrazine, azoxystrobin, bentazon, λ-cyhalothrin, penoxsulam and terbuthylazine) in sediment, macrophytes (algae and aquatic plants) and aquatic animals were developed and validated. The established methods were matrix-dependent and were based on Selective Pressurized Liquid Extraction (SPLE) followed by on-line Solid Phase Extraction and Ultra Performance Liquid Chromatography-tandem Mass Spectrometry (on-line SPE-UPLC-ESI-MS/MS). This cutting-edge research methodology uses a small amount of sample, is time saving and reduces the use of organic solvents in compliance with Green Chemistry principles. The analytical features were adequate for all compounds in all studied matrices. The established methodology was applied on real marine samples and no pesticide concentrations above their respective method quantification limits were measured in sediments or aquatic plants. However, terbuthylazine was found in the macroalgae Ulva spp. (108ngg(-1)dw) and all the prospected pesticides were measured above their respective method quantification limits in the bivalve Scrobicularia plana (atrazine: 48ngg(-1)dw, azoxystrobin: 64ngg(-1)dw, bentazon: 33ngg(-1)dw, λ-cyhalothrin: 2531ngg(-1)dw, penoxsulam: 50ngg(-1)dw, and terbuthylazine: 44ngg(-1)dw). PMID:27234845

  12. Advanced proteomic liquid chromatography

    SciTech Connect

    Xie, Fang; Smith, Richard D.; Shen, Yufeng

    2012-10-26

    Liquid chromatography coupled with mass spectrometry is the predominant platform used to analyze proteomics samples consisting of large numbers of proteins and their proteolytic products (e.g., truncated polypeptides) and spanning a wide range of relative concentrations. This review provides an overview of advanced capillary liquid chromatography techniques and methodologies that greatly improve separation resolving power and proteomics analysis coverage, sensitivity, and throughput.

  13. Liquid Chromatography in 1982.

    ERIC Educational Resources Information Center

    Freeman, David H.

    1982-01-01

    Reviews trends in liquid chromatography including apparatus, factors affecting efficient separation of a mixture (peak sharpness and speed), simplified problem-solving, adsorption, bonded phase chromatography, ion selectivity, and size exclusion. The current trend is to control chemical selectivity by the liquid phase. (Author/JN)

  14. Column Liquid Chromatography.

    ERIC Educational Resources Information Center

    Majors, Ronald E.; And Others

    1984-01-01

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

  15. Single-Step Fabrication of Computationally Designed Microneedles by Continuous Liquid Interface Production.

    PubMed

    Johnson, Ashley R; Caudill, Cassie L; Tumbleston, John R; Bloomquist, Cameron J; Moga, Katherine A; Ermoshkin, Alexander; Shirvanyants, David; Mecham, Sue J; Luft, J Christopher; DeSimone, Joseph M

    2016-01-01

    Microneedles, arrays of micron-sized needles that painlessly puncture the skin, enable transdermal delivery of medications that are difficult to deliver using more traditional routes. Many important design parameters, such as microneedle size, shape, spacing, and composition, are known to influence efficacy, but are notoriously difficult to alter due to the complex nature of microfabrication techniques. Herein, we utilize a novel additive manufacturing ("3D printing") technique called Continuous Liquid Interface Production (CLIP) to rapidly prototype sharp microneedles with tuneable geometries (size, shape, aspect ratio, spacing). This technology allows for mold-independent, one-step manufacturing of microneedle arrays of virtually any design in less than 10 minutes per patch. Square pyramidal CLIP microneedles composed of trimethylolpropane triacrylate, polyacrylic acid and photopolymerizable derivatives of polyethylene glycol and polycaprolactone were fabricated to demonstrate the range of materials that can be utilized within this platform for encapsulating and controlling the release of therapeutics. These CLIP microneedles effectively pierced murine skin ex vivo and released the fluorescent drug surrogate rhodamine.

  16. Single-Step Fabrication of Computationally Designed Microneedles by Continuous Liquid Interface Production.

    PubMed

    Johnson, Ashley R; Caudill, Cassie L; Tumbleston, John R; Bloomquist, Cameron J; Moga, Katherine A; Ermoshkin, Alexander; Shirvanyants, David; Mecham, Sue J; Luft, J Christopher; DeSimone, Joseph M

    2016-01-01

    Microneedles, arrays of micron-sized needles that painlessly puncture the skin, enable transdermal delivery of medications that are difficult to deliver using more traditional routes. Many important design parameters, such as microneedle size, shape, spacing, and composition, are known to influence efficacy, but are notoriously difficult to alter due to the complex nature of microfabrication techniques. Herein, we utilize a novel additive manufacturing ("3D printing") technique called Continuous Liquid Interface Production (CLIP) to rapidly prototype sharp microneedles with tuneable geometries (size, shape, aspect ratio, spacing). This technology allows for mold-independent, one-step manufacturing of microneedle arrays of virtually any design in less than 10 minutes per patch. Square pyramidal CLIP microneedles composed of trimethylolpropane triacrylate, polyacrylic acid and photopolymerizable derivatives of polyethylene glycol and polycaprolactone were fabricated to demonstrate the range of materials that can be utilized within this platform for encapsulating and controlling the release of therapeutics. These CLIP microneedles effectively pierced murine skin ex vivo and released the fluorescent drug surrogate rhodamine. PMID:27607247

  17. Single-Step Fabrication of Computationally Designed Microneedles by Continuous Liquid Interface Production

    PubMed Central

    Johnson, Ashley R.; Caudill, Cassie L.; Tumbleston, John R.; Bloomquist, Cameron J.; Moga, Katherine A.; Ermoshkin, Alexander; Shirvanyants, David; Mecham, Sue J.; Luft, J. Christopher; DeSimone, Joseph M.

    2016-01-01

    Microneedles, arrays of micron-sized needles that painlessly puncture the skin, enable transdermal delivery of medications that are difficult to deliver using more traditional routes. Many important design parameters, such as microneedle size, shape, spacing, and composition, are known to influence efficacy, but are notoriously difficult to alter due to the complex nature of microfabrication techniques. Herein, we utilize a novel additive manufacturing (“3D printing”) technique called Continuous Liquid Interface Production (CLIP) to rapidly prototype sharp microneedles with tuneable geometries (size, shape, aspect ratio, spacing). This technology allows for mold-independent, one-step manufacturing of microneedle arrays of virtually any design in less than 10 minutes per patch. Square pyramidal CLIP microneedles composed of trimethylolpropane triacrylate, polyacrylic acid and photopolymerizable derivatives of polyethylene glycol and polycaprolactone were fabricated to demonstrate the range of materials that can be utilized within this platform for encapsulating and controlling the release of therapeutics. These CLIP microneedles effectively pierced murine skin ex vivo and released the fluorescent drug surrogate rhodamine. PMID:27607247

  18. Advanced proteomic liquid chromatography

    PubMed Central

    Xie, Fang; Smith, Richard D.; Shen, Yufeng

    2012-01-01

    Liquid chromatography coupled with mass spectrometry is the predominant platform used to analyze proteomics samples consisting of large numbers of proteins and their proteolytic products (e.g., truncated polypeptides) and spanning a wide range of relative concentrations. This review provides an overview of advanced capillary liquid chromatography techniques and methodologies that greatly improve separation resolving power and proteomics analysis coverage, sensitivity, and throughput. PMID:22840822

  19. Selective Single-Step Separation of a Mixture of Three Metal Ions by a Triphasic Ionic-Liquid-Water-Ionic-Liquid Solvent Extraction System.

    PubMed

    Vander Hoogerstraete, Tom; Blockx, Jonas; De Coster, Hendrik; Binnemans, Koen

    2015-08-10

    In a conventional solvent extraction system, metal ions are distributed between two immiscible phases, typically an aqueous and an organic phase. In this paper, the proof-of-principle is given for the distribution of metal ions between three immiscible phases, two ionic liquid phases with an aqueous phase in between them. Three-liquid-phase solvent extraction allows separation of a mixture of three metal ions in a single step, whereas at least two steps are required to separate three metals in the case of two-liquid-phase solvent extraction. In the triphasic system, the lower organic phase is comprised of the ionic liquid betainium- or choline bis(trifluoromethylsulfonyl)imide, whereas the upper organic phase is comprised of the ionic liquid trihexyl(tetradecyl)phosphonium bis(trifluoromethylsulfonyl)imide. The triphasic system was used for the separation of a mixture of tin(II), yttrium(III), and scandium(III) ions. PMID:26178665

  20. Combination of Electromembrane Extraction and Liquid-Phase Microextraction in a Single Step: Simultaneous Group Separation of Acidic and Basic Drugs.

    PubMed

    Huang, Chuixiu; Seip, Knut Fredrik; Gjelstad, Astrid; Shen, Xiantao; Pedersen-Bjergaard, Stig

    2015-07-01

    Electromembrane extraction (EME) and liquid-phase microextraction (LPME) were combined in a single step for the first time to realize simultaneous and clear group separation of basic and acidic drugs. Using 2-nitrophenyl octyl ether as the supported liquid membrane (SLM) for EME and dihexyl ether as the SLM for LPME, basic and acidic drugs were extracted and separated simultaneously from a low pH sample by EME and LPME, respectively. After 15 min of extraction, basic drugs (citalopram and sertraline) were exhaustively extracted, whereas the recoveries for acidic drugs (ketoprofen and ibuprofen) were in the range of 76%-86%. Longer extraction time provided higher recoveries for the acidic drugs, but this somewhat deteriorated the group separation. Matrices effects from the coexisting acidic drugs/basic drugs were tested, and we observed that simultaneous EME/LPME was not affected by coexisting drugs at high concentration. This approach was further investigated from human plasma. Extraction recoveries were strongly dependent on dilution of plasma with buffer and on extraction time. Finally, this simultaneous EME/LPME approach was evaluated in combination with liquid chromatography (LC)-MS. The linearity ranges for the basic and acidic drugs were 10-600 ng/mL and 1-60 μg/mL, respectively, with R(2) > 0.997 for all analytes. The repeatability at three different levels for all analytes was less than 15%. The limits of quantification (LOQ, S/N = 10) were found to be 4.0-6.3 ng/mL and 0.6-0.9 μg/mL for basic and acidic drugs, respectively. Simultaneous EME/LPME enabled efficient group separation of basic and acidic analytes under optimum experimental conditions for both EME and LPME.

  1. High Performance Liquid Chromatography

    NASA Astrophysics Data System (ADS)

    Talcott, Stephen

    High performance liquid chromatography (HPLC) has many applications in food chemistry. Food components that have been analyzed with HPLC include organic acids, vitamins, amino acids, sugars, nitrosamines, certain pesticides, metabolites, fatty acids, aflatoxins, pigments, and certain food additives. Unlike gas chromatography, it is not necessary for the compound being analyzed to be volatile. It is necessary, however, for the compounds to have some solubility in the mobile phase. It is important that the solubilized samples for injection be free from all particulate matter, so centrifugation and filtration are common procedures. Also, solid-phase extraction is used commonly in sample preparation to remove interfering compounds from the sample matrix prior to HPLC analysis.

  2. High performance aptamer affinity chromatography for single-step selective extraction and screening of basic protein lysozyme.

    PubMed

    Han, Bin; Zhao, Chao; Yin, Junfa; Wang, Hailin

    2012-08-15

    A DNA aptamer based high-performance affinity chromatography is developed for selective extraction and screening of a basic protein lysozyme. First, a poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolithic column was synthesized in situ by thermally initiated radical polymerization, and then an anti-lysozyme DNA aptamer was covalently immobilized on the surface of the monolith through a 16-atom spacer arm. The target protein lysozyme but non-target proteins can be trapped by the immobilized anti-lysozyme DNA aptamer. In contrast, lysozyme cannot be trapped by the immobilized oligodeoxynucleotide that does not contain the sequence of the anti-lysozyme DNA aptamer. The study clearly demonstrates the trapping of lysozyme by the immobilized anti-lysozyme DNA aptamer is mainly due to specific recognition rather than simple electrostatic interaction of positively charged protein and the negatively charged DNA. The inter-day precision was determined as 0.8% for migration time and 4.2% for peak area, respectively. By the use of aptamer affinity monolith, a screening strategy is developed to selectively extract lysozyme from chicken egg white, showing the advantages of high efficiency, low cost and ease-of-operation.

  3. High-Performance Liquid Chromatography

    NASA Astrophysics Data System (ADS)

    Reuhs, Bradley L.; Rounds, Mary Ann

    High-performance liquid chromatography (HPLC) developed during the 1960s as a direct offshoot of classic column liquid chromatography through improvements in the technology of columns and instrumental components (pumps, injection valves, and detectors). Originally, HPLC was the acronym for high-pressure liquid chromatography, reflecting the high operating pressures generated by early columns. By the late 1970s, however, high-performance liquid chromatography had become the preferred term, emphasizing the effective separations achieved. In fact, newer columns and packing materials offer high performance at moderate pressure (although still high pressure relative to gravity-flow liquid chromatography). HPLC can be applied to the analysis of any compound with solubility in a liquid that can be used as the mobile phase. Although most frequently employed as an analytical technique, HPLC also may be used in the preparative mode.

  4. A simple and sensitive single-step method for gas chromatography-mass spectrometric determination of fipronil and its metabolites in sugarcane juice, jaggery and sugar.

    PubMed

    Ramasubramanian, Thirumalaiandi; Paramasivam, Mariappan; Jayanthi, Ramabhadran; Chandrasekaran, Subramaniam

    2014-05-01

    A simple and sensitive single-step method for gas chromatography-mass spectrometric determination of fipronil and its metabolites viz., fipronil desulfinyl, fipronil sulphide and fipronil sulphone in sugarcane juice, jaggery and sugar has been developed. Acetonitrile was superior to ethyl acetate in terms of selectivity, though they were on par with each other in terms of recoveries. This method does not require any cleanup as the PSA-based cleanup was on par with no-cleanup treatment. Interestingly, the recoveries of fipronil and its metabolites decreased with increased amounts of C18 from 10 to 50mg/g of matrix. Matrix effects were insignificant and the limit of quantification was 0.005μg/g. The recoveries of fipronil and its metabolites varied between 87.5% and 108.5% with the RSD of 0.2-5.3% for all the three matrices studied. This method has also been validated by monitoring fipronil and its metabolites in the retail outlet samples of sugarcane juice, jaggery and sugar.

  5. Determination of thiocyanate in saliva by headspace gas chromatography-mass spectrometry, following a single-step aqueous derivatization with triethyloxonium tetrafluoroborate.

    PubMed

    Ammazzini, Sara; Onor, Massimo; Pagliano, Enea; Mester, Zoltán; Campanella, Beatrice; Pitzalis, Emanuela; Bramanti, Emilia; D'Ulivo, Alessandro

    2015-06-26

    A novel method for the determination of salivary thiocyanate is presented. Thiocyanate was converted into ethyl thiocyanate by single-step aqueous derivatization based on triethyloxonium tetrafluoroborate and measured by gas chromatography-mass spectrometry (15 min runtime). The ethyl thiocyanate derivative is volatile and can be sampled from the headspace. The derivatization chemistry proposed allows for separation of the analyte from saliva matrix whose introduction in the measurement system is avoided. Quantitation of the analyte was obtained by isotope dilution, employing a (13)C-enriched thiocyanate as internal standard. Technical details and fundamental aspects of derivatization chemistry and calibration strategy are presented. The method was validated by comparison with a standard method based on ion chromatography. The two independent methodologies produced results in agreement within 3%. Also a three level spike recovery test was carried out for validation purpose and quantitative recoveries were attained. The method is fast, simple, safe, and sensitive. Measurement of a 1 mL volume 50 ng/g of thiocyanate standard produced a signal-to-noise ratio of 250 for the analytical peak. This method is therefore suitable for ultra-trace determination of thiocyanate (low part-per-billion range). For the application described the full detection potential of the method was not required and the sample preparation presented has been designed for quantitation of saliva samples containing 1-400 μg/g of thiocyanate with a combined standard uncertainty of 2% relative for saliva samples containing 25 μg/g of thiocyanate. This method was applied for the determination of thiocyanate in human saliva samples.

  6. Liquid-phase chromatography detector

    DOEpatents

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

    1983-11-08

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

  7. Liquid-phase chromatography detector

    DOEpatents

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

    1983-01-01

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

  8. High-performance liquid chromatography.

    PubMed

    Clevett, K J

    1990-01-01

    Gas chromatography has developed over the past 25 years or so into one of the most extensively used on-line analytical techniques in industrial process control and optimization. Liquid chromatography, and its several individual techniques, is firmly established in the laboratory, but its on-line process use has not developed as rapidly as GC. At the present time, only three companies (Applied Automation Inc., Dionex Corp., and Millipore Corp.) are active in this area. Nevertheless, substantial growth in on-line process LC is predicted for the next few years. The techniques of HPLC (normal-phase and reversed-phase), IEC, and SEC have great potential in industry as on-line analytical techniques, including the new field of biotechnology. Computer-based, multistream, multicomponent systems should find extensive use in pilot-plant investigations, where their ability to gather large amounts of data (on-line rather than by laboratory testing) could have important implications. In bioprocess control, undoubtedly the greatest challenge will come in the area of sample-handling technique. On-line chromatography has traditionally involved the sampling and conditioning of fairly conventional process gases and liquids. One exception is in the plastics and elastomers areas, where on-line SEC has been used for polymer MWD measurement. Here the sample is more difficult to handle, and some specialized techniques have been used. In biotechnology, we are treading new ground; nevertheless, it is hoped that some of the experience in sample handling gained in industry over the past 25 years will be of use in this new field.

  9. Liquid phase chromatography on microchips.

    PubMed

    Kutter, Jörg P

    2012-01-20

    Over the past twenty years, the field of microfluidics has emerged providing one of the main enabling technologies to realize miniaturized chemical analysis systems, often referred to as micro-Total Analysis Systems (uTAS), or, more generally, Lab-on-a-Chip Systems (LOC) [1,2]. While microfluidics was driven forward a lot from the engineering side, especially with respect to ink jet and dispensing technology, the initial push and interest from the analytical chemistry community was through the desire to develop miniaturized sensors, detectors, and, very early on, separation systems. The initial almost explosive development of, in particular, chromatographic separation systems on microchips, has, however, slowed down in recent years. This review takes a closer, critical look at how liquid phase chromatography has been implemented in miniaturized formats over the past several years, what is important to keep in mind when developing or working with separations in a miniaturized format, and what challenges and pitfalls remain.

  10. Mixed Stationary Liquid Phases for Gas-Liquid Chromatography.

    ERIC Educational Resources Information Center

    Koury, Albert M.; Parcher, Jon F.

    1979-01-01

    Describes a laboratory technique for use in an undergraduate instrumental analysis course that, using the interpretation of window diagrams, prepares a mixed liquid phase column for gas-liquid chromatography. A detailed procedure is provided. (BT)

  11. [High-performance liquid-liquid chromatography in beverage analysis].

    PubMed

    Bricout, J; Koziet, Y; de Carpentrie, B

    1978-01-01

    Liquid liquid chromatography was performed with columns packed with stationary phases chemically bonded to silica microparticules. These columns show a high efficiency and are used very easily. Flavouring compounds like aromatic aldehydes which have a low volatility were analyzed in brandy using a polar phase alkylnitrile. Sapid substances like amarogentin in Gentiana lutea or glyryrrhizin in Glycyrrhiza glabra were determined by reversed phase chromatography. Finally ionizable substances like synthetic dyes can be analyzed by paired ion chromatography witha non polar stationary phase.

  12. Liquid chromatography detection unit, system, and method

    SciTech Connect

    Derenzo, Stephen E.; Moses, William W.

    2015-10-27

    An embodiment of a liquid chromatography detection unit includes a fluid channel and a radiation detector. The radiation detector is operable to image a distribution of a radiolabeled compound as the distribution travels along the fluid channel. An embodiment of a liquid chromatography system includes an injector, a separation column, and a radiation detector. The injector is operable to inject a sample that includes a radiolabeled compound into a solvent stream. The position sensitive radiation detector is operable to image a distribution of the radiolabeled compound as the distribution travels along a fluid channel. An embodiment of a method of liquid chromatography includes injecting a sample that comprises radiolabeled compounds into a solvent. The radiolabeled compounds are then separated. A position sensitive radiation detector is employed to image distributions of the radiolabeled compounds as the radiolabeled compounds travel along a fluid channel.

  13. CATALYST ACTIVITY MAINTENANCE FOR THE LIQUID PHASE SYNTHESIS GAS-TO-DIMETHYL ETHER PROCESS PART II: DEVELOPMENT OF ALUMINUM PHOSPHATE AS THE DEHYDRATION CATALYST FOR THE SINGLE-STEP LIQUID PHASE SYNGAS-TO-DME PROCESS

    SciTech Connect

    Xiang-Dong Peng

    2002-05-01

    At the heart of the single-step liquid phase syngas-to-DME process (LPDME{trademark}) is a catalyst system that can be active as well as stable. In the Alternative Fuels I program, a dual-catalyst system containing a Cu-based commercial methanol synthesis catalyst (BASF S3-86) and a commercial dehydration material ({gamma}-alumina) was demonstrated. It provided the productivity and selectivity expected from the LPDME process. However, the catalyst system deactivated too rapidly to warrant a viable commercial process [1]. The mechanistic investigation in the early part of the DOE's Alternative Fuels II program revealed that the accelerated catalyst deactivation under LPDME conditions is due to detrimental interaction between the methanol synthesis catalyst and methanol dehydration catalyst [2,3]. The interaction was attributed to migration of Cu- and/or Zn-containing species from the synthesis catalyst to the dehydration catalyst. Identification of a dehydration catalyst that did not lead to this detrimental interaction while retaining adequate dehydration activity was elusive. Twenty-nine different dehydration materials were tested, but none showed the desired performance [2]. The search came to a turning point when aluminum phosphate was tested. This amorphous material is prepared by precipitating a solution containing Al(NO{sub 3}){sub 3} and H{sub 3}PO{sub 4} with NH{sub 4}OH, followed by washing, drying and calcination. The aluminum phosphate catalyst has adequate dehydration activity and good stability. It can co-exist with the Cu-based methanol synthesis catalyst without negatively affecting the latter catalyst's stability. This report documents the details of the development of this catalyst. These include initial leads, efforts in improving activity and stability, investigation and development of the best preparation parameters and procedures, mechanistic understanding and resulting preparation guidelines, and the accomplishments of this work.

  14. The effect of metal ions as co-catalysts on acidic ionic liquid catalyzed single-step saccharification of corn stover in water.

    PubMed

    Wiredu, Bernard; Amarasekara, Ananda S

    2015-01-01

    The effects of adding Cr(3+), Mn(2+), Fe(3+), Co(2+) Ni(2+), Cu(2+), Zn(2+) and La(3+) chlorides as co-catalysts to 1-(1-propylsulfonic)-3-methylimidazolium chloride acidic ionic liquid catalyzed saccharification of corn stover in aqueous medium was studied at 140-170 °C, by measuring the total reducing sugar (TRS) and glucose yields. The samples with Mn(2+), Fe(3+), Co(2+) as co-catalysts produced higher TRS yields compared to the sample without the metal ions. The Mn(2+) produced the highest catalytic effect enhancements and produced TRS yields of 68.0%, 72.9%, 90.2% and 87.9% at 140, 150, 160 and 170 °C respectively; whereas the corn stover samples without the Mn(2+) produced TRS yields of 42.9%, 52.3%, 54.4% and 53.5% at the same four temperatures. At higher temperatures of 160 and 170 °C, all metal ions studied produced significant enhancements in glucose yields, except Cr(3+). The addition of La(3+) as a co-catalyst produced the highest glucose yield improvement. PMID:25911191

  15. The effect of metal ions as co-catalysts on acidic ionic liquid catalyzed single-step saccharification of corn stover in water.

    PubMed

    Wiredu, Bernard; Amarasekara, Ananda S

    2015-01-01

    The effects of adding Cr(3+), Mn(2+), Fe(3+), Co(2+) Ni(2+), Cu(2+), Zn(2+) and La(3+) chlorides as co-catalysts to 1-(1-propylsulfonic)-3-methylimidazolium chloride acidic ionic liquid catalyzed saccharification of corn stover in aqueous medium was studied at 140-170 °C, by measuring the total reducing sugar (TRS) and glucose yields. The samples with Mn(2+), Fe(3+), Co(2+) as co-catalysts produced higher TRS yields compared to the sample without the metal ions. The Mn(2+) produced the highest catalytic effect enhancements and produced TRS yields of 68.0%, 72.9%, 90.2% and 87.9% at 140, 150, 160 and 170 °C respectively; whereas the corn stover samples without the Mn(2+) produced TRS yields of 42.9%, 52.3%, 54.4% and 53.5% at the same four temperatures. At higher temperatures of 160 and 170 °C, all metal ions studied produced significant enhancements in glucose yields, except Cr(3+). The addition of La(3+) as a co-catalyst produced the highest glucose yield improvement.

  16. Instrument platforms for nano liquid chromatography.

    PubMed

    Šesták, Jozef; Moravcová, Dana; Kahle, Vladislav

    2015-11-20

    The history of liquid chromatography started more than a century ago and miniaturization and automation are two leading trends in this field. Nanocolumn liquid chromatography (nano LC) and largely synonymous capillary liquid chromatography (capillary LC) are the most recent results of this process where miniaturization of column dimensions and sorbent particle size play crucial role. Very interesting results achieved in the research of extremely miniaturized LC columns at the end of the last century lacked distinctive raison d'être and only advances in mass spectrometry brought a real breakthrough. Configuration of nano LC-electrospray ionization mass spectrometry (LC-ESI-MS) has become a basic tool in bioanalytical chemistry, especially in proteomics. This review discusses and summarizes past and current trends in the realization of nano liquid chromatography (nano LC) platforms. Special attention is given to the mobile phase delivery under nanoflow rates (isocratic, gradient) and sample injection to the nanocolumn. Available detection techniques applied in nano LC separations are also briefly discussed. We followed up the key themes from the original scientific reports over gradual improvements up to the contemporary commercial solutions.

  17. Preparative isolation of the lectin jacalin by anion-exchange high-performance liquid chromatography.

    PubMed

    De Simone, S G; Santos, R; Araujo, M F; Pinho, R T

    1994-12-30

    The lectin jacalin from Artocarpus integrifolia was purified to homogeneity in a single step by preparative anion-exchange high-performance liquid chromatography (HPLC). Selection of the optimum chromatographic parameters in gradient elution allowed a rapid procedure to be obtained for the qualitative and quantitative isolation of the most important alpha- and alpha'-jacalin components. A recovery of 27-33% was obtained from a total soluble extract using a polyacrylate-DEAE HPLC column. The identities of the two isolated polypeptides were established by N-terminal amino acid sequence analysis and from the IgA1 binding lectin activity.

  18. High Performance Liquid Chromatography Experiments to Undergraduate Laboratories

    ERIC Educational Resources Information Center

    Kissinger, Peter T.; And Others

    1977-01-01

    Reviews the principles of liquid chromatography with electrochemical detection (LCEC), an analytical technique that incorporates the advantages of both liquids chromatography and electrochemistry. Also suggests laboratory experiments using this technique. (MLH)

  19. High perfomance liquid chromatography in pharmaceutical analyses.

    PubMed

    Nikolin, Branko; Imamović, Belma; Medanhodzić-Vuk, Saira; Sober, Miroslav

    2004-05-01

    In testing the pre-sale procedure the marketing of drugs and their control in the last ten years, high performance liquid chromatography replaced numerous spectroscopic methods and gas chromatography in the quantitative and qualitative analysis. In the first period of HPLC application it was thought that it would become a complementary method of gas chromatography, however, today it has nearly completely replaced gas chromatography in pharmaceutical analysis. The application of the liquid mobile phase with the possibility of transformation of mobilized polarity during chromatography and all other modifications of mobile phase depending upon the characteristics of substance which are being tested, is a great advantage in the process of separation in comparison to other methods. The greater choice of stationary phase is the next factor which enables realization of good separation. The separation line is connected to specific and sensitive detector systems, spectrafluorimeter, diode detector, electrochemical detector as other hyphernated systems HPLC-MS and HPLC-NMR, are the basic elements on which is based such wide and effective application of the HPLC method. The purpose high performance liquid chromatography (HPLC) analysis of any drugs is to confirm the identity of a drug and provide quantitative results and also to monitor the progress of the therapy of a disease.1) Measuring presented on the Fig. 1. is chromatogram obtained for the plasma of depressed patients 12 h before oral administration of dexamethasone. It may also be used to further our understanding of the normal and disease process in the human body trough biomedical and therapeutically research during investigation before of the drugs registration. The analyses of drugs and metabolites in biological fluids, particularly plasma, serum or urine is one of the most demanding but one of the most common uses of high performance of liquid chromatography. Blood, plasma or serum contains numerous endogenous

  20. Fast, comprehensive two-dimensional liquid chromatography

    PubMed Central

    Stoll, Dwight R.; Li, Xiaoping; Wang, Xiaoli; Carr, Peter W.; Porter, Sarah E. G.; Rutan, Sarah C.

    2011-01-01

    The absolute need to improve the separating power of liquid chromatography, especially for multi-constituent biological samples, is becoming increasingly evident. In response, over the past few years, there has been a great deal of interest in the development of two dimension liquid chromatography (2DLC). Just as 1DLC is preferred to 1DGC based on its compatibility with biological materials we believe that ultimately 2DLC will be preferred to the much more highly developed 2DGC for such samples. The huge advantage of 2D chromatographic techniques over 1D methods is inherent in the tremendous potential increase in peak capacity (resolving power). This is especially true of comprehensive 2D chromatography wherein it is possible, under ideal conditions, to obtain a total peak capacity equal to the product of the peak capacities of the first and second dimension separations. However, the very long timescale (typically several hours to tens of hours) of comprehensive 2DLC is clearly its chief drawback. Recent advances in the use of higher temperatures to speed up isocratic and gradient elution liquid chromatography have been used to decrease the time needed to do the second dimension LC separation of 2DLC to about 20 seconds for a full gradient elution run. Thus fast, high temperature LC is becoming a very promising technique. Peak capacities of over 2000 and rates of peak capacity production of nearly 1 peak/s have been achieved. In consequence, many real samples showing more than 200 peaks with signal to noise ratios of better than 10:1 have been run in total times of under 30 minutes. This report is not intended to be a comprehensive review of 2DLC, but is deliberately focused on the issues involved in doing fast 2DLC by means of elevating the column temperature; however, many issues of broader applicability will be discussed. PMID:17888443

  1. Csaba Horvath and preparative liquid chromatography

    SciTech Connect

    Guiochon, Georges A

    2005-07-01

    Few chromatographers have been interested in furthering preparative liquid chromatography. The pioneers, Tswett, Kuhn and Lederer, A.J.P. Martin, Tiselius, isolated fractions but as an intermediate step in the analysis of their samples. The progress in electronics and sensors, and in their miniaturization has lead to the paradoxical situation that the analysts never see the transient pure fractions that their detector quantitates. Yet, over the last 25 years, preparative liquid chromatography has become an important industrial process for the separation, the extraction, and/or the purification of many pharmaceuticals or pharmaceutical intermediates, including pure enantiomers, purified peptides and proteins, compounds that are manufactured at the relatively large industrial scale of a few kilograms to several hundred tons per year. This development that has strongly affected the modern pharmaceutical industry is mainly due to the pioneering work of Csaba Horvath. His work in preparative HPLC was critical at both the practical and the theoretical levels. He was the first scientist in modern times to pay serious attention to the relationships between the curvature of the equilibrium isotherms, the competitive nature of nonlinear isotherms, and the chromatographic band profiles of complex mixtures. The thermodynamics of multi-component phase equilibria and mass transfer kinetics in chromatography attracted his interest and were the focus of ground-breaking contributions. He investigated displacement chromatography, an old method invented by Tiselius that Csaba was first to implement in HPLC. This choice was explained by the essential characteristic of displacement chromatography, in that it delivers fractions that can be far more concentrated than the feed. Remarkably, once the basics of nonlinear chromatography had been mastered in his group, most of the applications that were studied by his coworkers dealt with peptides of various sizes and with proteins. Thus, all

  2. Spillage detector for liquid chromatography systems

    NASA Technical Reports Server (NTRS)

    Jarvis, M. J.; Fulton, D. S. (Inventor)

    1986-01-01

    A spillage detector device for use in conjunction with fractionation of liquid chromatography systems which includes a spillage recieving enclosure beneath the fractionation area is described. A sensing device having a plurality of electrodes of alternating polarity is mounted within the spillage recieving enclosure. Detection circuitry, responsive to conductivity between electrodes, is operatively connected to the sensing device. The detection circuitry feeds into the output circuitry. The output circuit has relaying and switching circuitry directed to a solenoid, an alarm system and a pump. The solenoid is connected to the pliable conduit of the chromatography system. The alarm system comprises an audio alarm and a visual signal. A 115-volt power system interconnected with the pump, the solenoid, the sensing device, and the detection and output circuitry.

  3. Multi-dimensional Liquid Chromatography in Proteomics

    PubMed Central

    Zhang, Xiang; Fang, Aiqin; Riley, Catherine P.; Wang, Mu; Regnier, Fred E.; Buck, Charles

    2010-01-01

    Proteomics is the large-scale study of proteins, particularly their expression, structures and functions. This still-emerging combination of technologies aims to describe and characterize all expressed proteins in a biological system. Because of upper limits on mass detection of mass spectrometers, proteins are usually digested into peptides and the peptides are then separated, identified and quantified from this complex enzymatic digest. The problem in digesting proteins first and then analyzing the peptide cleavage fragments by mass spectrometry is that huge numbers of peptides are generated that overwhelm direct mass spectral analyses. The objective in the liquid chromatography approach to proteomics is to fractionate peptide mixtures to enable and maximize identification and quantification of the component peptides by mass spectrometry. This review will focus on existing multidimensional liquid chromatographic (MDLC) platforms developed for proteomics and their application in combination with other techniques such as stable isotope labeling. We also provide some perspectives on likely future developments. PMID:20363391

  4. Trends in High Performance Liquid Chromatography for Cultural Heritage.

    PubMed

    Degano, Ilaria; La Nasa, Jacopo

    2016-04-01

    The separation, detection and quantitation of specific species contained in a sample in the field of Cultural Heritage requires selective, sensitive and reliable methods. Procedures based on liquid chromatography fulfil these requirements and offer a wide range of applicability in terms of analyte types and concentration range. The main applications of High Performance Liquid Chromatography in this field are related to the separation and detection of dyestuffs in archaeological materials and paint samples by reversed-phase liquid chromatography with suitable detectors. The relevant literature will be revised, with particular attention to sample treatment strategies and future developments. Reversed phase chromatography has also recently gained increasing importance in the analysis of lipid binders and lipid materials in archaeological residues: the main advantages and disadvantages of the new approaches will be discussed. Finally, the main applications of ion chromatography and size exclusion chromatography in the field of Cultural Heritage will be revised in this chapter. PMID:27573145

  5. Trends in High Performance Liquid Chromatography for Cultural Heritage.

    PubMed

    Degano, Ilaria; La Nasa, Jacopo

    2016-04-01

    The separation, detection and quantitation of specific species contained in a sample in the field of Cultural Heritage requires selective, sensitive and reliable methods. Procedures based on liquid chromatography fulfil these requirements and offer a wide range of applicability in terms of analyte types and concentration range. The main applications of High Performance Liquid Chromatography in this field are related to the separation and detection of dyestuffs in archaeological materials and paint samples by reversed-phase liquid chromatography with suitable detectors. The relevant literature will be revised, with particular attention to sample treatment strategies and future developments. Reversed phase chromatography has also recently gained increasing importance in the analysis of lipid binders and lipid materials in archaeological residues: the main advantages and disadvantages of the new approaches will be discussed. Finally, the main applications of ion chromatography and size exclusion chromatography in the field of Cultural Heritage will be revised in this chapter.

  6. Determination of starch lysophospholipids in rice using liquid chromatography-mass spectrometry (LC-MS).

    PubMed

    Liu, Lei; Tong, Chuan; Bao, Jinsong; Waters, Daniel L E; Rose, Terry J; King, Graham J

    2014-07-16

    Acquiring a complete understanding of rice starch lysophospholipids (LPLs), their biosynthetic pathways and genetic diversity, and the influence of genotype by environment interactions has been hampered by the lack of efficient high-throughput extraction and analysis methods. It was hypothesized a single-step aqueous n-propanol extraction combined with liquid chromatography-mass spectrometry (LC-MS) could be employed to analyze starch LPLs in white rice. The investigation found different grinding methods showed little effect on the final LPL detected, and a simple single-step extraction with 75% n-propanol (8 mL/0.15 g) heated at 100 °C for 2 h was as effective as an onerous multistep extraction method. A LC-MS method was optimized to simultaneously quantify 10 major LPLs in rice starch within 15 min. This method enables total and individual starch LPL analysis of a large number of rice samples at little cost. This approach could be applied to starch LPLs in other cereals.

  7. Use of high pressure liquid chromatography in the study of liquid lubricant oxidation

    NASA Technical Reports Server (NTRS)

    Morales, W.

    1982-01-01

    The general principles of classical liquid chromatography and high-pressure liquid chromatography (HPLC) are reviewed, and their advantages and disadvantages are compared. Several chromatographic techniques are reviewed, and the analysis of a C-ether liquid lubricant by each technique is illustrated. An analysis by size exclusion chromatography of an ester lubricant, which had been degraded using a micro-oxidation apparatus, is illustrated to show how HPLC can be used in the study of high-temperature lubricant degradation.

  8. Chromatography

    MedlinePlus

    Chromatography is a way of separating two or more chemical compounds. Chemical compounds are chemicals that are ... of chemical compound. There are different kinds of chromatography. These include gas, high pressure liquid, or ion ...

  9. Instrumentation for hand-portable liquid chromatography.

    PubMed

    Sharma, Sonika; Plistil, Alex; Simpson, Robert S; Liu, Kun; Farnsworth, Paul B; Stearns, Stanley D; Lee, Milton L

    2014-01-31

    Liquid chromatography (LC) has lagged behind gas chromatography (GC) in developments related to hand-portable instrumentation. In this work, a new battery-operated (24V DC) nano-flow pumping system with a stop-flow injector was developed and integrated with an on-column UV-absorption detector (254nm) that was reduced in size to an acceptable weight and power usage for field operation. The pumping system, which includes nano-flow pump, stepper motor and high-pressure valve weighs only 1.372kg (3lbs) and can generate up to 110.32MPa (16,000psi) pressure. A major advantage of this pump is that it does not employ a splitter, since it was specifically designed for capillary column use. The volume capacity of the pump is 24μL, and a sample volume as low as 10nL can be injected. Flow rate calibration (300nL to 6.12μL per min) was performed, and an accuracy >99.94% was obtained. The percent injection carry-over was found to be low (RSD 0.31%), which makes it practical for quantitative analysis. The detector linear range and limit of detection (LOD) were determined using sodium anthraquinone-2-sulfonate. A linear regression coefficient (R) of 0.9996 was obtained for a plot of log peak area versus log concentration over the range of 3.2μM to 6.5mM, and the LOD (S/N=3) was found to be 7.8fmol (0.13μM). The short term noise of the detector is comparable to commercially available detectors (∼10(-5)AU). In this work, the system was tested in the laboratory using regular line power (120V AC) with an AC to DC adapter. Reversed-phase isocratic separations were performed using a 15.5cm×75μm i.d. fused silica capillary column containing a monolithic stationary phase synthesized from 1,6-hexanediol dimethacrylate. Good retention time repeatability (RSD 0.09-0.74%) was obtained for a mixture containing an unretained marker (i.e., uracil) and a homologous series of alkyl benzenes.

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

    ERIC Educational Resources Information Center

    Bohman, Ove; And Others

    1982-01-01

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

  11. Ion Exchange and Liquid Column Chromatography.

    ERIC Educational Resources Information Center

    Walton, Harold F.

    1980-01-01

    Emphasizes recent advances in principles and methodology in ion exchange and chromatography. Two tables list representative examples for inorganic ions and organic compounds. Cites 544 references. (CS)

  12. Study of tanshinone IIA tissue distribution in rat by liquid chromatography-tandem mass spectrometry method.

    PubMed

    Bi, Hui-chang; Law, Francis C P; Zhong, Guo-ping; Xu, Chen-shu; Pan, Ying; Ding, Liang; Chen, Xiao; Zhao, Li-zi; Xu, Qiong; Huang, Min

    2007-05-01

    A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated for determining tanshinone IIA in rat tissues. After a single step liquid-liquid extraction with diethyl ether, tanshinone IIA and loratadine (internal standard) was subjected to LC/MS/MS analysis using positive electro-spray ionization under selected reaction monitoring mode. Chromatographic separation of tanshinone IIA and loratadine was achieved on a Hypersil BDS C(18) column (i.d. 2.1 x 50 mm, 5 microm) with a mobile phase consisting of methanol-1% formic acid (90:10, v/v) at a flow rate of 300 microL/min. The intra-day and inter-day precision of the method were less than 10.2 and 12.4%, respectively. The intra-day and inter-day accuracies ranged from 99.7 to 109.7%. The lowest limit of quantification for tanshinone IIA was 1 ng/mL. The method was applied to a tanshinone IIA tissue distribution study after an oral dose of 60 mg/kg to rats. Tanshinone IIA tissue concentrations decreased in the order of stomach > small intestine > lung > liver > fat > muscle > kidneys > spleen > heart > plasma > brain > testes. Tanshinone IIA still could be detected in most of the tissues at 20 h post-dosing. These results indicate that the LC/MS/MS method was rapid and sensitive to quantify tanshinone IIA in different rat tissues. PMID:17357178

  13. Micro-polarimeter for high performance liquid chromatography

    DOEpatents

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

    1985-01-01

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

  14. Performance of the same column in supercritical fluid chromatography and in liquid chromatography.

    PubMed

    Lambert, Nándor; Felinger, Attila

    2015-08-28

    We have studied the chromatographic behavior of the homologous series of alkylbenzenes (ranging from octylbenzene to octadecylbenzene) on the same C18 reversed-phase column in supercritical fluid chromatography (SFC) and reversed phase liquid chromatography (RPLC) at various experimental conditions, such as different eluent compositions, flow-rates, and mobile phase densities. The first and the second moments of the peaks were used to estimate the overall mass-transfer processes in both chromatographic modes using the stochastic model of chromatography. The results confirm that in SFC - as the density of the mobile phase is influenced by the flow-rate - there is a broader variation of mass-transfer properties than in liquid chromatography. As expected, the optimum mobile phase velocity is higher in SFC, but there is no real difference in the minimum value of plate height, i.e. in the optimum efficiency.

  15. Serum theophylline analysis by high-pressure liquid chromatography.

    PubMed

    Evenson, M A; Warren, B L

    1976-06-01

    We have developed and evaluated a rapid, high-pressure liquid-chromatographic method for theophylline in serum. Only 0.2 ml of serum is required for each determination, and the sensitivity of this method is 0.5 mg/liter. This method, involving liquid extraction and silica adsorption chromatography, provides adequate selectivity, accuracy, and precision for routine or research applications. Little sample preparation is required before chromatography. We found no endogenous or exogenous interferences. Use of beta-hydroxypropyl theophylline as the internal standard provides reproducible results for this micro-scale method.

  16. Determination of Sulfonamide Residues in Food by Micellar Liquid Chromatography

    PubMed Central

    Szymański, Arkadiusz

    2008-01-01

    This paper presents new methods of determination of sulfonamide residues in food products of animal origin, based on liquid chromatography with a micellar mobile phase. The methods employ a technique of direct injection of the sample and preliminary isolation of the analyte by extraction in the liquid-solid and liquid-liquid system. The methods have been characterized by providing the parameters of the calibration curves, the range of linearity, limit of detection, and precision and accuracy of particular determinations. The recovery of the sulfonamides introduced into the food products studied has also been determined. PMID:19696937

  17. Liquid Chromatography Applied to Space System

    NASA Astrophysics Data System (ADS)

    Poinot, Pauline; Chazalnoel, Pascale; Geffroy, Claude; Sternberg, Robert; Carbonnier, Benjamin

    Searching for signs of past or present life in our Solar System is a real challenge that stirs up the curiosity of scientists. Until now, in situ instrumentation was designed to detect and determine concentrations of a wide number of organic biomarkers. The relevant method which was and still is employed in missions dedicated to the quest of life (from Viking to ExoMars) corresponds to the pyrolysis-GC-MS. Along the missions, this approach has been significantly improved in terms of extraction efficiency and detection with the use of chemical derivative agents (e.g. MTBSTFA, DMF-DMA, TMAH…), and in terms of analysis sensitivity and resolution with the development of in situ high-resolution mass spectrometer (e.g. TOF-MS). Thanks to such an approach, organic compounds such as amino acids, sugars, tholins or polycyclic aromatic hydrocarbons (PAHs) were expected to be found. However, while there’s a consensus that the GC-MS of Viking, Huygens, MSL and MOMA space missions worked the way they had been designed to, pyrolysis is much more in debate (Glavin et al. 2001; Navarro-González et al. 2006). Indeed, (1) it is thought to remove low levels of organics, (2) water and CO2 could interfere with the detection of likely organic pyrolysis products, and (3) only low to mid-molecular weight organic molecules can be detected by this technique. As a result, researchers are now focusing on other in situ techniques which are no longer based on the volatility of the organic matter, but on the liquid phase extraction and analysis. In this line, micro-fluidic systems involving sandwich and/or competitive immunoassays (e.g. LMC, SOLID; Parro et al. 2005; Sims et al. 2012), micro-chip capillary electrophoreses (e.g. MOA; Bada et al. 2008), or nanopore-based analysis (e.g. BOLD; Schulze-Makuch et al. 2012) have been conceived for in situ analysis. Thanks to such approaches, molecular biological polymers (polysaccharides, polypeptides, polynucleotides, phospholipids, glycolipids

  18. Liquid Chromatography Applied to Space System

    NASA Astrophysics Data System (ADS)

    Poinot, Pauline; Chazalnoel, Pascale; Geffroy, Claude; Sternberg, Robert; Carbonnier, Benjamin

    Searching for signs of past or present life in our Solar System is a real challenge that stirs up the curiosity of scientists. Until now, in situ instrumentation was designed to detect and determine concentrations of a wide number of organic biomarkers. The relevant method which was and still is employed in missions dedicated to the quest of life (from Viking to ExoMars) corresponds to the pyrolysis-GC-MS. Along the missions, this approach has been significantly improved in terms of extraction efficiency and detection with the use of chemical derivative agents (e.g. MTBSTFA, DMF-DMA, TMAH…), and in terms of analysis sensitivity and resolution with the development of in situ high-resolution mass spectrometer (e.g. TOF-MS). Thanks to such an approach, organic compounds such as amino acids, sugars, tholins or polycyclic aromatic hydrocarbons (PAHs) were expected to be found. However, while there’s a consensus that the GC-MS of Viking, Huygens, MSL and MOMA space missions worked the way they had been designed to, pyrolysis is much more in debate (Glavin et al. 2001; Navarro-González et al. 2006). Indeed, (1) it is thought to remove low levels of organics, (2) water and CO2 could interfere with the detection of likely organic pyrolysis products, and (3) only low to mid-molecular weight organic molecules can be detected by this technique. As a result, researchers are now focusing on other in situ techniques which are no longer based on the volatility of the organic matter, but on the liquid phase extraction and analysis. In this line, micro-fluidic systems involving sandwich and/or competitive immunoassays (e.g. LMC, SOLID; Parro et al. 2005; Sims et al. 2012), micro-chip capillary electrophoreses (e.g. MOA; Bada et al. 2008), or nanopore-based analysis (e.g. BOLD; Schulze-Makuch et al. 2012) have been conceived for in situ analysis. Thanks to such approaches, molecular biological polymers (polysaccharides, polypeptides, polynucleotides, phospholipids, glycolipids

  19. Liquid Chromatography-Mass Spectrometry-based Quantitative Proteomics

    SciTech Connect

    Xie, Fang; Liu, Tao; Qian, Weijun; Petyuk, Vladislav A.; Smith, Richard D.

    2011-07-22

    Liquid chromatography-mass spectrometry (LC-MS)-based quantitative proteomics has become increasingly applied for a broad range of biological applications due to growing capabilities for broad proteome coverage and good accuracy in quantification. Herein, we review the current LC-MS-based quantification methods with respect to their advantages and limitations, and highlight their potential applications.

  20. Multichannel Detection in High-Performance Liquid Chromatography.

    ERIC Educational Resources Information Center

    Miller, James C.; And Others

    1982-01-01

    A linear photodiode array is used as the photodetector element in a new ultraviolet-visible detection system for high-performance liquid chromatography (HPLC). Using a computer network, the system processes eight different chromatographic signals simultaneously in real-time and acquires spectra manually/automatically. Applications in fast HPLC…

  1. Quantification of Tea Flavonoids by High Performance Liquid Chromatography

    ERIC Educational Resources Information Center

    Freeman, Jessica D.; Niemeyer, Emily D.

    2008-01-01

    We have developed a laboratory experiment that uses high performance liquid chromatography (HPLC) to quantify flavonoid levels in a variety of commercial teas. Specifically, this experiment analyzes a group of flavonoids known as catechins, plant-derived polyphenolic compounds commonly found in many foods and beverages, including green and black…

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

    PubMed

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

    1977-08-01

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

  3. Determination of Caffeine in Beverages by High Performance Liquid Chromatography.

    ERIC Educational Resources Information Center

    DiNunzio, James E.

    1985-01-01

    Describes the equipment, procedures, and results for the determination of caffeine in beverages by high performance liquid chromatography. The method is simple, fast, accurate, and, because sample preparation is minimal, it is well suited for use in a teaching laboratory. (JN)

  4. An Inexpensive Liquid Chromatography Apparatus for Undergraduate Teaching.

    ERIC Educational Resources Information Center

    McCamish, Malcolm; And Others

    1982-01-01

    Describes an inexpensive, low-pressure liquid chromatography pump, slurry filler, stainless steel columns, and injector system suitable for the undergraduate laboratory or routine analysis. Includes sectional diagram of the pump and construction diagram of the preparative columns. (Author/SK)

  5. LCEC: The Combination of Liquid Chromatography and Electrochemistry.

    ERIC Educational Resources Information Center

    Kissinger, Peter T.

    1983-01-01

    Use of combined liquid chromatography and finite-current electrochemistry (LCEC) procedures are discussed. Also discusses the relationship between electroactivity and molecular structure, selectivity in LCEC, and LCEC applications. Because of its selectivity and low detection limits, the procedures are most often applied in biomedical and…

  6. Mallow carotenoids determined by high-performance liquid chromatography

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mallow (corchorus olitorius) is a green vegetable, which is widely consumed either fresh or dry by Middle East population. This study was carried out to determine the contents of major carotenoids quantitatively in mallow, by using a High Performance Liquid Chromatography (HPLC) equipped with a Bis...

  7. High-Performance Liquid Chromatography-Mass Spectrometry.

    ERIC Educational Resources Information Center

    Vestal, Marvin L.

    1984-01-01

    Reviews techniques for online coupling of high-performance liquid chromatography with mass spectrometry, emphasizing those suitable for application to nonvolatile samples. Also summarizes the present status, strengths, and weaknesses of various techniques and discusses potential applications of recently developed techniques for combined liquid…

  8. Sample injector for high pressure liquid chromatography

    DOEpatents

    Paul, Phillip H.; Arnold, Don W.; Neyer, David W.

    2001-01-01

    Apparatus and method for driving a sample, having a well-defined volume, under pressure into a chromatography column. A conventional high pressure sampling valve is replaced by a sample injector composed of a pair of injector components connected in series to a common junction. The injector components are containers of porous dielectric material constructed so as to provide for electroosmotic flow of a sample into the junction. At an appropriate time, a pressure pulse from a high pressure source, that can be an electrokinetic pump, connected to the common junction, drives a portion of the sample, whose size is determined by the dead volume of the common junction, into the chromatographic column for subsequent separation and analysis. The apparatus can be fabricated on a substrate for microanalytical applications.

  9. Analysis of pesticide multi-residues in leafy vegetables by ultrasonic solvent extraction and liquid chromatography-tandem mass spectrometry.

    PubMed

    Pan, Jian; Xia, Xiao-Xiao; Liang, Juan

    2008-01-01

    Ultrasonic solvent extraction (USE) of pesticide multi-residues including monocrotophos, dimethoate, imidacloprid, carbendazim, carbaryl and simazine from leafy vegetables is presented. The extraction procedure was optimized with regard to the solvent type and amount, sonication time and number of extraction steps. The extract did not need clean-up before injected into liquid chromatography-tandem mass spectrometry (LC-MS-MS) which was employed together with electron microscope to verify the effect of USE method. The proposed procedure allows the extraction of six pesticide residues in a single step with 40 ml of ethyl acetate for 35 min sonication, providing recovery over 83% and LOQ less than 1.4 microg/kg. The optimized USE method is a simple, low cost and an effective preparation method for determination of pesticide multi-residues at trace levels in leafy vegetables in comparison with homogenized extraction method. PMID:17664080

  10. 21 CFR 862.2250 - Gas liquid chromatography system for clinical use.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Gas liquid chromatography system for clinical use... Instruments § 862.2250 Gas liquid chromatography system for clinical use. (a) Identification. A gas liquid chromatography system for clinical use is a device intended to separate one or more drugs or compounds from...

  11. 21 CFR 862.2250 - Gas liquid chromatography system for clinical use.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Gas liquid chromatography system for clinical use... Instruments § 862.2250 Gas liquid chromatography system for clinical use. (a) Identification. A gas liquid chromatography system for clinical use is a device intended to separate one or more drugs or compounds from...

  12. 21 CFR 862.2250 - Gas liquid chromatography system for clinical use.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Gas liquid chromatography system for clinical use... Instruments § 862.2250 Gas liquid chromatography system for clinical use. (a) Identification. A gas liquid chromatography system for clinical use is a device intended to separate one or more drugs or compounds from...

  13. 21 CFR 862.2250 - Gas liquid chromatography system for clinical use.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Gas liquid chromatography system for clinical use... Instruments § 862.2250 Gas liquid chromatography system for clinical use. (a) Identification. A gas liquid chromatography system for clinical use is a device intended to separate one or more drugs or compounds from...

  14. 21 CFR 862.2260 - High pressure liquid chromatography system for clinical use.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false High pressure liquid chromatography system for... Clinical Laboratory Instruments § 862.2260 High pressure liquid chromatography system for clinical use. (a) Identification. A high pressure liquid chromatography system for clinical use is a device intended to...

  15. 21 CFR 862.2250 - Gas liquid chromatography system for clinical use.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Gas liquid chromatography system for clinical use... Instruments § 862.2250 Gas liquid chromatography system for clinical use. (a) Identification. A gas liquid chromatography system for clinical use is a device intended to separate one or more drugs or compounds from...

  16. 21 CFR 862.2260 - High pressure liquid chromatography system for clinical use.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false High pressure liquid chromatography system for... Clinical Laboratory Instruments § 862.2260 High pressure liquid chromatography system for clinical use. (a) Identification. A high pressure liquid chromatography system for clinical use is a device intended to...

  17. 21 CFR 862.2260 - High pressure liquid chromatography system for clinical use.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false High pressure liquid chromatography system for... Clinical Laboratory Instruments § 862.2260 High pressure liquid chromatography system for clinical use. (a) Identification. A high pressure liquid chromatography system for clinical use is a device intended to...

  18. 21 CFR 862.2260 - High pressure liquid chromatography system for clinical use.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false High pressure liquid chromatography system for... Clinical Laboratory Instruments § 862.2260 High pressure liquid chromatography system for clinical use. (a) Identification. A high pressure liquid chromatography system for clinical use is a device intended to...

  19. 21 CFR 862.2260 - High pressure liquid chromatography system for clinical use.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false High pressure liquid chromatography system for... Clinical Laboratory Instruments § 862.2260 High pressure liquid chromatography system for clinical use. (a) Identification. A high pressure liquid chromatography system for clinical use is a device intended to...

  20. Exploring Fluorous Affinity by Liquid Chromatography.

    PubMed

    Catani, Martina; Guzzinati, Roberta; Marchetti, Nicola; Pasti, Luisa; Cavazzini, Alberto

    2015-07-01

    Terms such as "fluorous affinity" and "fluorophilicity" have been used to describe the unique partition and sorption properties often exhibited by highly fluorinated organic compounds, that is molecules rich in sp(3) carbon-fluorine bonds. In this work, we made use of a highly fluorinated stationary phase and a series of benzene derivatives to study the effect of one single perfluorinated carbon on the chromatographic behavior and adsorption properties of molecules. For this purpose, the adsorption equilibria of α,α,α-trifluorotoluene, toluene, and other alkylbenzenes have been studied by means of nonlinear chromatography in a variety of acetonitrile/water eluents. Our results reveal that one single perfluorinated carbon is already enough to induce a drastic change in the adsorption properties of molecules on the perfluorinated stationary phase. In particular, it has been found that adsorption is monolayer if the perfluoroalkyl carbon is present but that, when this unit is missing, molecules arrange as multilayer stack structures. These findings can contribute to the understanding of molecular mechanisms of fluorous affinity. PMID:26047527

  1. Optimizing heterosurface adsorbent synthesis for liquid chromatography

    NASA Astrophysics Data System (ADS)

    Bogoslovskii, S. Yu.; Serdan, A. A.

    2016-03-01

    The structural and geometric parameters of a silica matrix (SM) for the synthesis of heterosurface adsorbents (HAs) are optimized. Modification is performed by shielding the external surfaces of alkyl-modified silica (AS) using human serum albumin and its subsequent crosslinking. The structural and geometric characteristics of the SM, AS, and HA are measured via low-temperature nitrogen adsorption. It is found that the structural characteristics of AS pores with diameters D < 6 nm do not change during HA synthesis, while the volume of pores with diameters of 6 nm < D < 9 nm shrinks slightly due to the adsorption of albumin in the pore orifices. It is established that the volume of pores with diameters D > 9 nm reduces significantly due to adsorption of albumin. It is concluded that silica gel with a maximum pore size distribution close to 5 nm and a minimal proportion of pores with D > 9 nm is optimal for HA synthesis; this allows us to achieve the greatest similarity between the chromatographic retention parameters for HA and AS. The suitability of the synthesized adsorbents for analyzing drugs in biological fluids through direct sample injection is confirmed by chromatography. It was found that the percentage of the protein fraction detected at the outlet of the chromatographic column is 98%.

  2. Ionic liquids as surfactants in micellar liquid chromatography.

    PubMed

    Flieger, Jolanta; Siwek, Agata; Pizoń, Magdalena; Czajkowska-Żelazko, Anna

    2013-05-01

    This paper is devoted to application of ionic liquids as surfactants in LC of organic compounds, derivatives of 1,4-thiosemicarbazides. According to HPLC requirements the most advantageous conditions such as transparency for ultraviolet light, low CMC, additional inorganic salt additives, and appropriate organic solvent were established. The CMC was determined using conductivity measurements. Suitability of two different stationary phases: RP-C18 and cyanopropyl bonded phase was examined under micellar conditions. Chosen ionic liquid surfactant was compared to common traditional amphiphilic reagent - SDS. Elaborated on chromatographic micellar conditions were tested as a pilot technique for prediction of distribution coefficients of organic analytes in ionic liquid-based aqueous two-phase system. PMID:23609988

  3. A newcomer's guide to nano-liquid-chromatography of peptides.

    PubMed

    Fröhlich, Thomas; Arnold, Georg J

    2009-01-01

    LC-MS/MS is one of the most powerful techniques in the field of proteomics allowing high throughput identification of proteins out of complex protein mixtures. Besides high sample throughput, the analytical sensitivity is one of the major benefits of this technology. A prerequisite for sensitive LC-MS/MS approaches is chromatography with very low flow rates in the nanoliter per minute range, usually referred to as nano-liquid chromatography (nano-LC). However, to perform this separation technology, an appropriate instrumental setup as well experienced operators are a prerequisite. The aim of this chapter is to help nano-LC newcomers to get introduced to this fascinating technology. Technical components of nano-LC systems like solvent delivery systems, sample injection systems, and nano-chromatography columns are described. Detailed procedures to mount, test, and operate the system are outlined, and advices for an effective troubleshooting are provided.

  4. Standard Flow Liquid Chromatography for Shotgun Proteomics in Bioenergy Research

    PubMed Central

    González Fernández-Niño, Susana M.; Smith-Moritz, A. Michelle; Chan, Leanne Jade G.; Adams, Paul D.; Heazlewood, Joshua L.; Petzold, Christopher J.

    2015-01-01

    Over the past 10 years, the bioenergy field has realized significant achievements that have encouraged many follow on efforts centered on biosynthetic production of fuel-like compounds. Key to the success of these efforts has been transformational developments in feedstock characterization and metabolic engineering of biofuel-producing microbes. Lagging far behind these advancements are analytical methods to characterize and quantify systems of interest to the bioenergy field. In particular, the utilization of proteomics, while valuable for identifying novel enzymes and diagnosing problems associated with biofuel-producing microbes, is limited by a lack of robustness and limited throughput. Nano-flow liquid chromatography coupled to high-mass accuracy, high-resolution mass spectrometers has become the dominant approach for the analysis of complex proteomic samples, yet such assays still require dedicated experts for data acquisition, analysis, and instrument upkeep. The recent adoption of standard flow chromatography (ca. 0.5 mL/min) for targeted proteomics has highlighted the robust nature and increased throughput of this approach for sample analysis. Consequently, we assessed the applicability of standard flow liquid chromatography for shotgun proteomics using samples from Escherichia coli and Arabidopsis thaliana, organisms commonly used as model systems for lignocellulosic biofuels research. Employing 120 min gradients with standard flow chromatography, we were able to routinely identify nearly 800 proteins from E. coli samples; while for samples from Arabidopsis, over 1,000 proteins could be reliably identified. An examination of identified peptides indicated that the method was suitable for reproducible applications in shotgun proteomics. Standard flow liquid chromatography for shotgun proteomics provides a robust approach for the analysis of complex samples. To the best of our knowledge, this study represents the first attempt to validate the standard

  5. Standard flow liquid chromatography for shotgun proteomics in bioenergy research.

    PubMed

    González Fernández-Niño, Susana M; Smith-Moritz, A Michelle; Chan, Leanne Jade G; Adams, Paul D; Heazlewood, Joshua L; Petzold, Christopher J

    2015-01-01

    Over the past 10 years, the bioenergy field has realized significant achievements that have encouraged many follow on efforts centered on biosynthetic production of fuel-like compounds. Key to the success of these efforts has been transformational developments in feedstock characterization and metabolic engineering of biofuel-producing microbes. Lagging far behind these advancements are analytical methods to characterize and quantify systems of interest to the bioenergy field. In particular, the utilization of proteomics, while valuable for identifying novel enzymes and diagnosing problems associated with biofuel-producing microbes, is limited by a lack of robustness and limited throughput. Nano-flow liquid chromatography coupled to high-mass accuracy, high-resolution mass spectrometers has become the dominant approach for the analysis of complex proteomic samples, yet such assays still require dedicated experts for data acquisition, analysis, and instrument upkeep. The recent adoption of standard flow chromatography (ca. 0.5 mL/min) for targeted proteomics has highlighted the robust nature and increased throughput of this approach for sample analysis. Consequently, we assessed the applicability of standard flow liquid chromatography for shotgun proteomics using samples from Escherichia coli and Arabidopsis thaliana, organisms commonly used as model systems for lignocellulosic biofuels research. Employing 120 min gradients with standard flow chromatography, we were able to routinely identify nearly 800 proteins from E. coli samples; while for samples from Arabidopsis, over 1,000 proteins could be reliably identified. An examination of identified peptides indicated that the method was suitable for reproducible applications in shotgun proteomics. Standard flow liquid chromatography for shotgun proteomics provides a robust approach for the analysis of complex samples. To the best of our knowledge, this study represents the first attempt to validate the standard

  6. Liquid chromatography and mass spectrometry in food allergen detection.

    PubMed

    Fæste, Christiane Kruse; Rønning, Helene Thorsen; Christians, Uwe; Granum, Per Einar

    2011-02-01

    Food allergy is an important issue in the field of food safety because of the hazards for affected persons and the hygiene requirements and legal regulations imposed on the food industry. Consumer protection and law enforcement require suitable analytical techniques for the detection of allergens in foods. Immunological methods are currently preferred; however, confirmatory alternatives are needed. The determination of allergenic proteins by liquid chromatography and mass spectrometry has greatly advanced in recent years, and gel-free allergenomics is becoming a routinely used approach for the identification and quantitation of food allergens. The present review provides a brief overview of the principles of proteomic procedures, various chromatographic set ups, and mass spectrometry instrumentation used in allergenomics. A compendium of published liquid chromatography methods, proteomic analyses, typical marker peptides, and quantitative assays for 14 main allergy-causing foods is also included.

  7. An electrostatic precipitator for preparative gas-liquid chromatography.

    PubMed

    Borka, L; Privett, O S

    1966-03-01

    The effect of the operating variables of electrostatic precipitators on the recovery and structure of methyl esters and related aerosol forming compounds collected in preparative gas-liquid chromatography was studied.Aerosol formation was prevented by AC or DC voltages of 5000 to 12000 volts. AC was more effective than DC but caused changes in structure which were detectable by both thin-layer and gas-liquid chromatographic methods of analysis.An apparatus of simple construction and operation was designed for the collection of methyl esters and its use demonstrated with several model compounds.

  8. Determination of serotonin released from coffee wax by liquid chromatography.

    PubMed

    Kele, M; Ohmacht, R

    1996-04-12

    A simple hydrolysis and extraction method was developed for the release of serotonin (5-hydroxytryptamine) from a coffee wax sample obtained from decaffeination of coffee beans. The recoverable amount of serotonin was determined by reversed-phase high-performance liquid chromatography with gradient elution and UV detection, using the standard addition method. Different type of basic deactivated chromatographic columns were used for the separation.

  9. Nitrogen-sensitive thermionic detection in microcolumn liquid chromatography.

    PubMed

    Gluckman, J C; Novotny, M

    1985-10-01

    The dual-flame thermionic detector for microcolumn liquid chromatography has been improved and optimized for nitrogen sensitivity. The total column effluent is concentrically nebulized and aspirated directly into an air-hydrogen diffusion flame, while detection limits of 1.4 X 10(-11) g nitrogen/sec at the maximum of a Gaussian peak are achieved. Detection linearity spans three orders of magnitude. An example of the analysis of underivatized barbiturate standards is provided.

  10. Determination of serotonin released from coffee wax by liquid chromatography.

    PubMed

    Kele, M; Ohmacht, R

    1996-04-12

    A simple hydrolysis and extraction method was developed for the release of serotonin (5-hydroxytryptamine) from a coffee wax sample obtained from decaffeination of coffee beans. The recoverable amount of serotonin was determined by reversed-phase high-performance liquid chromatography with gradient elution and UV detection, using the standard addition method. Different type of basic deactivated chromatographic columns were used for the separation. PMID:8680597

  11. Characterisation of poly(vinyl alcohol) by liquid chromatography techniques

    SciTech Connect

    Meehan, E.; Warner, F.P.; Patterson, M.

    1995-12-01

    The molecular weight distribution of poly (vinyl alcohol) can be measured by aqueous size exclusion chromatography methods but the choice of eluent is critical in eliminating non size exclusion behavior. Aqueous size exclusion experiments have been carried out using a number of eluents including standard electrolytes and surfactants. The most favorable molecular size separation was obtained using 0.25% w/v sodium lauryl sulphate as eluent. Compositional distributions in copolymer systems can be assessed using high performance liquid chromatography employing a reverse phase separation mechanism. For poly (vinyl alcohol) gradient elution with water/tetrahydrofuran was found to produce a separation according to composition. Fast gradient elution (>10% tetrahydrofuran/minute) suggested abroad distribution of composition which was verified using a column packed with non-porous beads. Slower gradient elution (<1% tetrahydrofuran/minute) suggested that this was not due to a gradual composition change but rather discrete fractions of similarly hydrophobic material.

  12. Mass transfer mechanism in chiral reversed phase liquid chromatography.

    PubMed

    Gritti, Fabrice; Guiochon, Georges

    2014-03-01

    The mechanism of mass transfer in chiral chromatography was investigated using an experimental protocol already applied in RPLC and HILIC chromatography. The different contributions to the reduced height equivalent to a theoretical plate (HETP) include the longitudinal diffusion HETP term, the solid-liquid mass transfer resistance HETP term, the short-range eddy dispersion HETP term, and the long-range eddy dispersion HETP term. Their accurate measurement permits the determination of the adsorption rate constant kads of trans-stilbene enantiomers on a column packed with Lux 5 μm Cellulose-1 particles. The experimental results demonstrate that the number of adsorption-desorption steps per unit time of chiral compounds on polysaccharide-based chiral stationary phases is four orders of magnitude smaller than that of achiral compounds.

  13. Analysis of radioactive mixed hazardous waste using derivatization gas chromatography/mass spectrometry, liquid chromatography, and liquid chromatography/mass spectrometry

    SciTech Connect

    Campbell, J.A.; Lerner, B.D.; Bean, R.M.; Grant, K.E.; Lucke, R.B.; Mong, G.M.; Clauss, S.A.

    1994-08-01

    Six samples of core segments from Tank 101-SY were analyzed for chelators, chelator fragments, and several carboxylic acids by derivatization gas chromatography/mass spectrometry. The major components detected were ethylenediaminetetraacetic acid, nitroso-iminodiacetic acid, nitrilotriacetic acid, citric acid, succinic acid, and ethylenediaminetriacetic acid. The chelator of highest concentration was ethylenediaminetetraacetic acid in all six samples analyzed. Liquid chromatography was used to quantitate low molecular weight acids including oxalic, formic, glycolic, and acetic acids, which are present in the waste as acid salts. From 23 to 61% of the total organic carbon in the samples analyzed was accounted for by these acids.

  14. Modeling of closed-loop recycling liquid-liquid chromatography: Analytical solutions and model analysis.

    PubMed

    Kostanyan, Artak E

    2015-08-01

    In closed-loop recycling (CLR) chromatography, the effluent from the outlet of a column is directly returned into the column through the sample feed line and continuously recycled until the required separation is reached. To select optimal operating conditions for the separation of a given feed mixture, an appropriate mathematical description of the process is required. This work is concerned with the analysis of models for the CLR separations. Due to the effect of counteracting mechanisms on separation of solutes, analytical solutions of the models could be helpful to understand and optimize chromatographic processes. The objective of this work was to develop analytical expressions to describe the CLR counter-current (liquid-liquid) chromatography (CCC). The equilibrium dispersion and cell models were used to describe the transport and separation of solutes inside a CLR CCC column. The Laplace transformation is applied to solve the model equations. Several possible CLR chromatography methods for the binary and complex mixture separations are simulated.

  15. Simultaneous determination of buprenorphine, norbuprenorphine and naloxone in human plasma by liquid chromatography/tandem mass spectrometry.

    PubMed

    Liu, Yongzhen; Li, Xiaohua; Xu, Allan; Nasser, Azmi F; Heidbreder, Christian

    2016-02-20

    A simple, sensitive and rapid liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated for simultaneous quantification of naloxone, buprenorphine and its metabolite norbuprenorphine in human plasma. Human plasma samples were extracted using a single step liquid-liquid extraction, and then separated on an Imtakt Unison UK-C18 column (2.1×50mm, 3μm) using alkaline mobile phases with gradient elution. All of the analytes were detected in positive ion mode using multiple reaction monitoring (MRM). The method was validated and the specificity, linearity, lower limit of quantitation, precision, accuracy, recoveries and stability were determined. The linear range was 20-10000pg/mL for buprenorphine and norbuprenorphine; and 1-500pg/mL for naloxone. The correlation coefficient (R(2)) values for all three analytes were ≥0.995. The precision and accuracy for intra-day and inter-day were <11.0%. The recoveries were >63% and matrix effects were tracked by the deuterated internal standards (IS) with the IS-normalized matrix factor ranging from 0.96 to 1.33 for all three analytes. The validated method was successfully applied in a clinical pharmacokinetic study with low dose administration of sublingual buprenorphine and naloxone.

  16. Simultaneous determination of tanshinone IIA and cryptotanshinone in rat plasma by liquid chromatography-electrospray ionisation-mass spectrometry.

    PubMed

    Li, Jing; Wang, Guangji; Li, Peng; Hao, Haiping

    2005-11-01

    Tanshinone IIA (TS) and cryptotanshinone (CT) are the major active constituents contained in Radix salvia miltiorrhiza. This paper described a rapid, sensitive and specific assay for the simultaneous quantitative determination of TS and CT in rat plasma. After a single step of liquid-liquid extraction, plasma samples were analyzed by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) using a reversed-phase C18 column (150 mmx2.0 mm, 5 microm, Shim-pack VP-ODS column). The assay was linear in the concentration range of 2-200 ng/ml. The lower limits of quantification of TS and CT were 1 and 0.2 ng/ml, respectively. Recoveries of TS and CT were greater than 80%. The precisions and accuracies determined from 5 days were all within 12%. The assay was applied to a pharmacokinetic study in rats after an oral administration of total tanshinones with a dose of 150 mg/kg (containing 12% of TS and CT). Results showed that this simple and rapid method was sensitive enough to follow the plasma levels of TS and CT in rats, even though the concentration maximums of both were below 20 ng/ml after an oral administration of total tanshinones. PMID:16140046

  17. Simultaneous determination of tanshinone IIA and its three hydroxylated metabolites by liquid chromatography/tandem mass spectrometry.

    PubMed

    Li, Peng; Wang, Guang-Ji; Li, Jing; Hao, Hai-Ping; Zheng, Chao-Nan

    2006-01-01

    A rapid and sensitive method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the simultaneous determination of tanshinone IIA and its three hydroxylated metabolites, tanshinone IIB, hydroxytanshinone IIA and przewaquinone A, in a rat liver microsome was developed and fully validated. A single step of liquid-liquid extraction with ethyl acetate was utilized in this method. Chromatographic separation of the sample matrix from the analytes and the internal standard diazepam was performed using a Shim-pack VP-ODS analytical column. Detection was performed on a triple quadrupole tandem mass spectrometer equipped with an electrospray ionization source and operated in selected reaction monitoring (SRM) mode. The method was linear in the concentration range of 1-500 ng/mL for all analytes. The intra- and inter-day precisions (RSD %) were within 15% and deviations of the assay accuracies were within 15.0% for all analytes. The analytes proved to be stable during sample storage, preparation and analyses. This validated method was successfully applied to the enzyme kinetic study of tanshinone IIA in liver microsome. The elimination of tanshinone IIA and formation of tanshinone IIB and hydroxytanshinone IIA in the liver microsome all exhibited a sigmoidal kinetics profile. The formation of przewaquinone A shows a typical hyperbolic profile. In addition, this method has now been applied in the analysis of other bio-samples including plasma, urine, bile and feces. PMID:16470728

  18. Simultaneous determination of buprenorphine, norbuprenorphine and naloxone in human plasma by liquid chromatography/tandem mass spectrometry.

    PubMed

    Liu, Yongzhen; Li, Xiaohua; Xu, Allan; Nasser, Azmi F; Heidbreder, Christian

    2016-02-20

    A simple, sensitive and rapid liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated for simultaneous quantification of naloxone, buprenorphine and its metabolite norbuprenorphine in human plasma. Human plasma samples were extracted using a single step liquid-liquid extraction, and then separated on an Imtakt Unison UK-C18 column (2.1×50mm, 3μm) using alkaline mobile phases with gradient elution. All of the analytes were detected in positive ion mode using multiple reaction monitoring (MRM). The method was validated and the specificity, linearity, lower limit of quantitation, precision, accuracy, recoveries and stability were determined. The linear range was 20-10000pg/mL for buprenorphine and norbuprenorphine; and 1-500pg/mL for naloxone. The correlation coefficient (R(2)) values for all three analytes were ≥0.995. The precision and accuracy for intra-day and inter-day were <11.0%. The recoveries were >63% and matrix effects were tracked by the deuterated internal standards (IS) with the IS-normalized matrix factor ranging from 0.96 to 1.33 for all three analytes. The validated method was successfully applied in a clinical pharmacokinetic study with low dose administration of sublingual buprenorphine and naloxone. PMID:26730511

  19. Separation of sulfoalkylated cyclodextrins with hydrophilic interaction liquid chromatography.

    PubMed

    Jaramillo, Michael; Kirschner, Daniel L; Dai, Zhipeng; Green, Thomas K

    2013-11-01

    Determination of the charged state distributions of partially- and fully-substituted sulfoalkylated cyclodextrins was achieved using hydrophilic interaction liquid chromatography (HILIC). HILIC analysis of a spiked sample of the partially sulfopropylated cyclodextrins was achieved using a gradient to baseline resolve the charged states from -1 to -14. The fully-substituted CDs yielded a major peak with some trace impurities and the partially-substituted sulfopropylated cyclodextrins showed a wide range of charge states present in the mixture. Small changes in the structure of the cyclodextrins have a significant impact on the retention times of the various types of cyclodextrins investigated.

  20. Gas-liquid chromatography in lunar organic analysis.

    NASA Technical Reports Server (NTRS)

    Gehrke, C. W.

    1972-01-01

    Gas-liquid chromatography (GLC) is a powerful and sensitive method for the separation and detection of organic compounds at nanogram levels. The primary requirement for successful analyses is that the compounds of interest must be volatile under the chromatographic conditions employed. Nonvolatile organic compounds must be converted to volatile derivatives prior to analysis. The derivatives of choice must be both amenable to chromatographic separation and be relatively stable. The condition of volatility necessitates the development of efficient derivatization reactions for important groups of compounds as amino acids, carbohydrates, nucleosides, etc. Trimethylsilylation and trifluoroacetylation represent specific areas of recent prominence. Some relevant practical aspects of GLC are discussed.

  1. Size distributions of gold nanoclusters studied by liquid chromatography

    SciTech Connect

    WILCOXON,JESS P.; MARTIN,JAMES E.; PROVENCIO,PAULA P.

    2000-05-23

    The authors report high pressure liquid chromatography, (HPLC), and transmission electron microscopy, (TEM), studies of the size distributions of nanosize gold clusters dispersed in organic solvents. These metal clusters are synthesized in inverse micelles at room temperature and those investigated range in diameter from 1--10 nm. HPLC is sensitive enough to discern changes in hydrodynamic volume corresponding to only 2 carbon atoms of the passivating agent or metal core size changes of less than 4 {angstrom}. The authors have determined for the first time how the total cluster volume (metal core + passivating organic shell) changes with the size of the passivating agent.

  2. A novel amide stationary phase for hydrophilic interaction liquid chromatography and ion chromatography.

    PubMed

    Shen, Guobin; Zhang, Feifang; Yang, Bingcheng; Chu, Changhu; Liang, Xinmiao

    2013-10-15

    A novel amide stationary phase (ASP) for hydrophilic interaction liquid chromatography (HILIC) has been prepared via the Click chemistry method. It was based on the strategy that the amino group of Asparagine was easily transferred to the corresponding azido group and then clicked onto terminal alkyne-silica gel in the presence of Cu(I)-based catalyst. For the tested polar compounds including nucleosides and nucleic acid bases, ASP-based column has demonstrated good performance in terms of separation efficiency and column stability, and the retention mechanism was found to match well the typical HILIC retention. In addition, the ASP described here showed much better selectivity in separation of inorganic anions under ion chromatography mode relative to other kinds of commercial ASP.

  3. A novel amide stationary phase for hydrophilic interaction liquid chromatography and ion chromatography.

    PubMed

    Shen, Guobin; Zhang, Feifang; Yang, Bingcheng; Chu, Changhu; Liang, Xinmiao

    2013-10-15

    A novel amide stationary phase (ASP) for hydrophilic interaction liquid chromatography (HILIC) has been prepared via the Click chemistry method. It was based on the strategy that the amino group of Asparagine was easily transferred to the corresponding azido group and then clicked onto terminal alkyne-silica gel in the presence of Cu(I)-based catalyst. For the tested polar compounds including nucleosides and nucleic acid bases, ASP-based column has demonstrated good performance in terms of separation efficiency and column stability, and the retention mechanism was found to match well the typical HILIC retention. In addition, the ASP described here showed much better selectivity in separation of inorganic anions under ion chromatography mode relative to other kinds of commercial ASP. PMID:24054569

  4. Fiber-based monolithic columns for liquid chromatography.

    PubMed

    Ladisch, Michael; Zhang, Leyu

    2016-10-01

    Fiber-based monoliths for use in liquid chromatographic separations are defined by columns packed with aligned fibers, woven matrices, or contiguous fiber structures capable of achieving rapid separations of proteins, macromolecules, and low molecular weight components. A common denominator and motivating driver for this approach, first initiated 25 years ago, was reducing the cost of bioseparations in a manner that also reduced residence time of retained components while achieving a high ratio of mass to momentum transfer. This type of medium, when packed into a liquid chromatography column, minimized the fraction of stagnant liquid and resulted in a constant plate height for non-adsorbing species. The uncoupling of dispersion from eluent flow rate enabled the surface chemistry of the stationary phase to be considered separately from fluid transport phenomena and pointed to new ways to apply chemistry for the engineering of rapid bioseparations. This paper addresses developments and current research on fiber-based monoliths and explains how the various forms of this type of chromatographic stationary phase have potential to provide new tools for analytical and preparative scale separations. The different stationary phases are discussed, and a model that captures the observed constant plate height as a function of mobile phase velocity is reviewed. Methods that enable hydrodynamically stable fiber columns to be packed and operated over a range of mobile phase flow rates, together with the development of new fiber chemistries, are shown to provide columns that extend the versatility of liquid chromatography using monoliths, particularly at the preparative scale. Graphical Abstract Schematic representation of a sample mixture being separated by a rolled-stationary phase column, resulting separated peaks shown in the chromatogram. PMID:27553948

  5. Identification of polychlorinated styrene compounds in heron tissues by gas-liquid chromatography-mass spectrometry

    USGS Publications Warehouse

    Reichel, W.L.; Prouty, R.M.; Gay, M.L.

    1977-01-01

    Unknown compounds detected in Ardea herodias tissues are identified by gas-liquid chromatography-mass spectrometry as residues of octachlorostyrene. Heptachlorostyrene and hexachlorostyrene were tentatively identified.

  6. General theory of peak compression in liquid chromatography.

    PubMed

    Gritti, Fabrice

    2016-02-12

    A new and general expression of the peak compression factor in liquid chromatography is derived. It applies to any type of gradients induced by non-uniform columns (stationary) or by temporal variations (dynamic) of the elution strength related to changes in solvent composition, temperature, or in any external field. The new equation is validated in two ideal cases for which the exact solutions are already known. From a practical viewpoint, it is used to predict the achievable degree of peak compression for curved retention models, retained solvent gradients, and for temperature-programmed liquid chromatography. The results reveal that: (1) curved retention models affect little the compression factor with respect to the best linear strength retention models, (2) gradient peaks can be indefinitely compressed with respect to isocratic peaks if the propagation speed of the gradient (solvent or temperature) becomes smaller than the chromatographic velocity, (3) limitations are inherent to the maximum intensity of the experimental intrinsic gradient steepness, and (4) dynamic temperature gradients can be advantageously combined to solvent gradients in order to improve peak capacities of microfluidic separation devices.

  7. Purification of flavonoids from licorice using an off-line preparative two-dimensional normal-phase liquid chromatography/reversed-phase liquid chromatography method.

    PubMed

    Fan, Yunpeng; Fu, Yanhui; Fu, Qing; Cai, Jianfeng; Xin, Huaxia; Dai, Mei; Jin, Yu

    2016-07-01

    An orthogonal (71.9%) off-line preparative two-dimensional normal-phase liquid chromatography/reversed-phase liquid chromatography method coupled with effective sample pretreatment was developed for separation and purification of flavonoids from licorice. Most of the nonflavonoids were firstly removed using a self-made Click TE-Cys (60 μm) solid-phase extraction. In the first dimension, an industrial grade preparative chromatography was employed to purify the crude flavonoids. Click TE-Cys (10 μm) was selected as the stationary phase that provided an excellent separation with high reproducibility. Ethyl acetate/ethanol was selected as the mobile phase owing to their excellent solubility for flavonoids. Flavonoids co-eluted in the first dimension were selected for further purification using reversed-phase liquid chromatography. Multiple compounds could be isolated from one normal-phase fraction and some compounds with bad resolution in one-dimensional liquid chromatography could be prepared in this two-dimensional system owing to the orthogonal separation. Moreover, this two-dimensional liquid chromatography method was beneficial for the preparation of relatively trace flavonoid compounds, which were enriched in the first dimension and further purified in the second dimension. Totally, 24 flavonoid compounds with high purity were obtained. The results demonstrated that the off-line two-dimensional liquid chromatography method was effective for the preparative separation and purification of flavonoids from licorice.

  8. Enantioselective high performance liquid chromatography and supercritical fluid chromatography separation of spirocyclic terpenoid flavor compounds.

    PubMed

    Schaffrath, Mathias; Weidmann, Verena; Maison, Wolfgang

    2014-10-10

    Chiral spirocyclic terpenoids are abundant natural flavors with significant impact particularly on the food industry. Chromatographic methods for analytical and preparative separation of these compounds are therefore of high interest to natural product chemists in academia and industry. Gas chromatography on chiral stationary phases is currently the standard method for the separation of volatile terpenoids, limiting the scale to analytical quantities. We report herein high performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) protocols for the chiral separation of several racemic spirocyclic terpenoids such as the important flavors theaspirane and vitispirane. A screening of mobile phases and 16 commercially available chiral stationary phases (CSPs) largely based on polysaccharides led to identification of protocols for the separation of all terpenoids tested. SFC methods were found to be particularly useful for the separation of these spirocyclic flavors due to the volatility and low polarity of the compounds. The reported chiral HPLC and SFC protocols are scalable alternatives to gas chromatographic separations of volatile terpenoid flavors.

  9. Comparison of liquid chromatography-microchip/mass spectrometry to conventional liquid chromatography-mass spectrometry for the analysis of steroids.

    PubMed

    Ahonen, Linda; Keski-Rahkonen, Pekka; Saarelainen, Taija; Paviala, Jenni; Ketola, Raimo A; Auriola, Seppo; Poutanen, Matti; Kostianen, Risto

    2012-04-01

    The feasibility of a microfluidic-based liquid chromatography-electrospray ionization/mass spectrometric system (HPLC-Chip/ESI/MS) was studied and compared to a conventional narrow-bore liquid chromatography-electrospray ionization/mass spectrometric (LC-ESI/MS) system for the analysis of steroids. The limits of detection (LODs) for oxime derivatized steroids, expressed as concentrations, were slightly higher with the HPLC-Chip/MS system (50-300 pM) using an injection volume of 0.5 μL than with the conventional LC-ESI/MS (10-150 pM) using an injection volume of 40 μL. However, when the LODs are expressed as injected amounts, the sensitivity of the HPLC-Chip/MS system was about 50 times higher than with the conventional LC-ESI/MS system. The results indicate that the use of HPLC-Chip/MS system is clearly advantageous only in the analysis of low-volume samples. Both methods showed good linearity and good quantitative and chromatographic repeatability. In addition to the instrument comparisons with oxime derivatized steroids, the feasibility of the HPLC-Chip/MS system in the analysis of non-derivatized and oxime derivatized steroids was compared. The HPLC-Chip/MS method developed for non-derivatized steroids was also applied to the quantitative analysis of 15 mouse plasma samples.

  10. Evaluation between ultrahigh pressure liquid chromatography and high-performance liquid chromatography analytical methods for characterizing natural dyestuffs.

    PubMed

    Serrano, Ana; van Bommel, Maarten; Hallett, Jessica

    2013-11-29

    An evaluation was undertaken of ultrahigh pressure liquid chromatography (UHPLC) in comparison to high-performance liquid chromatography (HPLC) for characterizing natural dyes in cultural heritage objects. A new UHPLC method was optimized by testing several analytical parameters adapted from prior UHPLC studies developed in diverse fields of research. Different gradient elution programs were tested on seven UHPLC columns with different dimensions and stationary phase compositions by applying several mobile phases, flow rates, temperatures, and runtimes. The UHPLC method successfully provided more improved data than that achieved by the HPLC method. Indeed, even though carminic acid has shown circa 146% higher resolution with HPLC, UHPLC resulted in an increase of 41-61% resolution and a decrease of 91-422% limit of detection, depending on the dye compound. The optimized method was subsequently assigned to analyse 59 natural reference materials, in which 85 different components were ascribed with different physicochemical properties, in order to create a spectral database for future characterization of dyes in cultural heritage objects. The majority of these reference samples could be successfully distinguished with one single method through the examination of these compounds' retention times and their spectra acquired with a photodiode array detector. These results demonstrate that UHPLC analyses are extremely valuable for the acquisition of more precise chromatographic information concerning natural dyes with complex mixtures of different and/or closely related physicochemical properties, essential for distinguishing similar species of plants and animals used to colour cultural heritage objects. PMID:24139502

  11. Evaluation between ultrahigh pressure liquid chromatography and high-performance liquid chromatography analytical methods for characterizing natural dyestuffs.

    PubMed

    Serrano, Ana; van Bommel, Maarten; Hallett, Jessica

    2013-11-29

    An evaluation was undertaken of ultrahigh pressure liquid chromatography (UHPLC) in comparison to high-performance liquid chromatography (HPLC) for characterizing natural dyes in cultural heritage objects. A new UHPLC method was optimized by testing several analytical parameters adapted from prior UHPLC studies developed in diverse fields of research. Different gradient elution programs were tested on seven UHPLC columns with different dimensions and stationary phase compositions by applying several mobile phases, flow rates, temperatures, and runtimes. The UHPLC method successfully provided more improved data than that achieved by the HPLC method. Indeed, even though carminic acid has shown circa 146% higher resolution with HPLC, UHPLC resulted in an increase of 41-61% resolution and a decrease of 91-422% limit of detection, depending on the dye compound. The optimized method was subsequently assigned to analyse 59 natural reference materials, in which 85 different components were ascribed with different physicochemical properties, in order to create a spectral database for future characterization of dyes in cultural heritage objects. The majority of these reference samples could be successfully distinguished with one single method through the examination of these compounds' retention times and their spectra acquired with a photodiode array detector. These results demonstrate that UHPLC analyses are extremely valuable for the acquisition of more precise chromatographic information concerning natural dyes with complex mixtures of different and/or closely related physicochemical properties, essential for distinguishing similar species of plants and animals used to colour cultural heritage objects.

  12. Method transfer from high-pressure liquid chromatography to ultra-high-pressure liquid chromatography. II. Temperature and pressure effects.

    PubMed

    Åsberg, Dennis; Samuelsson, Jörgen; Leśko, Marek; Cavazzini, Alberto; Kaczmarski, Krzysztof; Fornstedt, Torgny

    2015-07-01

    The importance of the generated temperature and pressure gradients in ultra-high-pressure liquid chromatography (UHPLC) are investigated and compared to high-pressure liquid chromatography (HPLC). The drug Omeprazole, together with three other model compounds (with different chemical characteristics, namely uncharged, positively and negatively charged) were used. Calculations of the complete temperature profile in the column at UHPLC conditions showed, in our experiments, a temperature difference between the inlet and outlet of 16 °C and a difference of 2 °C between the column center and the wall. Through van't Hoff plots, this information was used to single out the decrease in retention factor (k) solely due to the temperature gradient. The uncharged solute was least affected by temperature with a decrease in k of about 5% while for charged solutes the effect was more pronounced, with k decreases up to 14%. A pressure increase of 500 bar gave roughly 5% increase in k for the uncharged solute, while omeprazole and the other two charged solutes gave about 25, 20 and 15% increases in k, respectively. The stochastic model of chromatography was applied to estimate the dependence of the average number of adsorption/desorption events (n) and the average time spent by a molecule in the stationary phase (τs) on temperature and pressure on peak shape for the tailing, basic solute. Increasing the temperature yielded an increase in n and decrease in τs which resulted in less skew at high temperatures. With increasing pressure, the stochastic modeling gave interesting results for the basic solute showing that the skew of the peak increased with pressure. The conclusion is that pressure effects are more pronounced for both retention and peak shape than the temperature effects for the polar or charged compounds in our study.

  13. Single-Step Purification of Ovalbumin from Egg White Using Aqueous Biphasic Systems

    PubMed Central

    Pereira, Matheus M.; Cruz, Rafaela A. P.; Almeida, Mafalda R.; Lima, Álvaro S.; Coutinho, João A. P.; Freire, Mara G.

    2016-01-01

    The ability of aqueous biphasic systems (ABS) composed of polyethylene glycols of different molecular weights (PEG 400, 600 and 1000) and buffered aqueous solutions of potassium citrate/citric acid (pH = 5.0 - 8.0) to selectively extract ovalbumin from egg white was here investigated. Phase diagrams, tie-lines and tie-line lengths were determined at 25ºC and the partitioning of ovalbumin in these systems was then evaluated. Aiming at optimizing the selective extraction of ovalbumin in the studied ABS, factors such as pH, PEG molecular weight and amount of the phase-forming components were initially investigated with pure commercial ovalbumin. In almost all ABS, it was observed a preferential partitioning of ovalbumin to the polymer-rich phase, with extraction efficiencies higher than 90%. The best ABS were then applied in the purification of ovalbumin from the real egg white matrix. In order to ascertain on the ovalbumin purity and yield, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion high performance liquid chromatography (SE-HPLC) analyses were conducted, confirming that the isolation/purification of ovalbumin from egg white was completely achieved in a single-step with a recovery yield of 65%. The results obtained show that polymer-salt-based ABS allow the selective extraction of ovalbumin from egg white with a simpler approach and better performance than previously reported. Finally, it is shown that ovalbumin can be completely recovered from the PEG-rich phase by an induced precipitation using an inexpensive and sustainable separation platform which can be easily applied on an industrial scale. PMID:27642253

  14. Separation of organophosphorus pesticides by using nano-liquid chromatography.

    PubMed

    Buonasera, Katia; D'Orazio, Giovanni; Fanali, Salvatore; Dugo, Paola; Mondello, Luigi

    2009-05-01

    In the present research, the separation of a series of organophosphorus pesticides (fensulfothion, fenamiphos, profenofos, fonofos, isofenphos, dialifos, sulprofos and prothiofos), by using nano-liquid chromatography (nano-LC) with UV detection is described. Three 100 microm ID capillary columns, packed with different silica-based stationary phases (CN, C(18), and phenyl), were investigated. Among these, the phenyl column offered the best results in terms of chromatographic performance, and was selected for pesticide analyses. Parameters, such as sample dilution solvent, injection volume, mobile phase composition and flow rate, were optimized in order to define the ideal experimental conditions. With the aim of improving sensitivity, on-column focusing of large injection volumes was applied: a sensitivity increase of circa 100-fold was attained, with limits of detection (LODs) and quantification (LOQs) within the 4.4-37.5 and 14.5-125.0 ng/mL ranges, respectively. The method was validated, with satisfactory results, through the measurement of the following parameters: limits of detection and quantification, precision, linearity and recovery. Finally, five different baby foods, previously fortified with a solution of the eight aforementioned pesticides, and then subjected to liquid-liquid extraction and solid-phase extraction clean-up, were analyzed. PMID:19321170

  15. Analysis of quality of aviation lubricating oils by means of liquid and gas-liquid chromatography

    SciTech Connect

    Kholostova, G.G.; Bakunin, V.N.; Shimonaev, G.S.

    1987-01-01

    The authors examine the basic methodological aspects of chromatographic analysis of the quality of oils for aircraft gas turbine engines, and certain relationships in oil aging that have been established on this basis. A commercial ester (designated PEE) was selected for investigation of pentaerythritol and C/sub 5/-C/sub 9/ synthetic fatty acids (SFA) which serves as the synthetic base stock for a number of aviation oils. The changes in PEE composition upon oxidation, with or without additives, were evaluated by means of gas-liquid chromatography in a Tsvet-100 chromatograph with a flame ionization detector. The results from examination of the original and oxidized PEE samples by means of gas and liquid chromatography are presented.

  16. Liquid chromatography-mass spectrometry in metabolomics research: mass analyzers in ultra high pressure liquid chromatography coupling.

    PubMed

    Forcisi, Sara; Moritz, Franco; Kanawati, Basem; Tziotis, Dimitrios; Lehmann, Rainer; Schmitt-Kopplin, Philippe

    2013-05-31

    The present review gives an introduction into the concept of metabolomics and provides an overview of the analytical tools applied in non-targeted metabolomics with a focus on liquid chromatography (LC). LC is a powerful analytical tool in the study of complex sample matrices. A further development and configuration employing Ultra-High Pressure Liquid Chromatography (UHPLC) is optimized to provide the largest known liquid chromatographic resolution and peak capacity. Reasonably UHPLC plays an important role in separation and consequent metabolite identification of complex molecular mixtures such as bio-fluids. The most sensitive detectors for these purposes are mass spectrometers. Almost any mass analyzer can be optimized to identify and quantify small pre-defined sets of targets; however, the number of analytes in metabolomics is far greater. Optimized protocols for quantification of large sets of targets may be rendered inapplicable. Results on small target set analyses on different sample matrices are easily comparable with each other. In non-targeted metabolomics there is almost no analytical method which is applicable to all different matrices due to limitations pertaining to mass analyzers and chromatographic tools. The specifications of the most important interfaces and mass analyzers are discussed. We additionally provide an exemplary application in order to demonstrate the level of complexity which remains intractable up to date. The potential of coupling a high field Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (ICR-FT/MS), the mass analyzer with the largest known mass resolving power, to UHPLC is given with an example of one human pre-treated plasma sample. This experimental example illustrates one way of overcoming the necessity of faster scanning rates in the coupling with UHPLC. The experiment enabled the extraction of thousands of features (analytical signals). A small subset of this compositional space could be mapped into a mass

  17. Sheath liquid effects in capillary high-performance liquid chromatography-electrospray mass spectrometry of oligonucleotides.

    PubMed

    Huber, C G; Krajete, A

    2000-02-18

    Fused-silica capillary columns of 200 microm inner diameter were packed with micropellicular, octadecylated, 2.3 microm poly(styrene-divinylbenzene) particles and applied to the separation of oligonucleotides by ion-pair reversed-phase high-performance liquid chromatography. Oligonucleotides were eluted at 50 degrees C with gradients of 3-13% acetonitrile in 50 mM triethylammonium bicarbonate. Addition of sheath liquid to the column effluent allowed the detection of oligonucleotides by electrospray ionization mass spectrometry using full-scan data acquisition with a detectability comparable to that obtained with UV detection. The signal-to-noise ratios with different sheath liquids increased in the order isopropanolliquid was found to influence the charge state distribution of oligonucleotides longer than 20 nucleotide units whereas no significant effect was observed with shorter oligonucleotides. Organic acids and bases in the sheath liquid generally deteriorated the signal-to-noise ratios in the chromatograms and mass spectra mainly because of increased background noise. Only a few charge states were observed in the mass spectra of oligonucleotides because of charge state reduction due to the presence of carbonic acid in the eluent. With triethylammonium hydrogencarbonate as chromatographic eluent and acetonitrile as sheath liquid, very few cation adducts of oligonucleotides were observed in the mass spectra. However, the presence of small amounts of monopotassium adducts enabled the calculation of the charge state of multiply charged ions. With acetonitrile as sheath liquid, 710 amol of a 16-mer oligonucleotide were detected using selected ion monitoring data acquisition with a signal-to-noise ratio of 3:1. Finally, capillary ion-pair reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry was

  18. Investigating the Retention Mechanisms of Liquid Chromatography Using Solid-Phase Extraction Cartridges

    ERIC Educational Resources Information Center

    O'Donnell, Mary E.; Musial, Beata A.; Bretz, Stacey Lowery; Danielson, Neil D.; Ca, Diep

    2009-01-01

    Liquid chromatography (LC) experiments for the undergraduate analytical laboratory course often illustrate the application of reversed-phase LC to solve a separation problem, but rarely compare LC retention mechanisms. In addition, a high-performance liquid chromatography instrument may be beyond what some small colleges can purchase. Solid-phase…

  19. Recent advances in ultra-high performance liquid chromatography for the analysis of traditional chinese medicine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional Chinese medicines (TCMs) have been widely used for the prevention and treatment of various diseases for thousands of years in China. Ultra Performance Liquid Chromatography (UHPLC) is a relatively new technique offering new possibilities in liquid chromatography. This paper reviews recen...

  20. Ribbon storage techniques for liquid chromatography: mass spectrometry

    SciTech Connect

    Smith, R.D.; Johnson, A.L.

    1981-06-01

    A new liquid chromatography-mass spectrometry (LC-MS) has been developed which allows semipermanent storage of the chromatographically separated material on a moving ribbon permitting multiple temperature analyses of a single LC separation. The new interface removes the major disadvantage of conventional moving ribbon devices by allowing analysis of a single LC separation at several different temperatures. Reconstructed ion chromatograms for the molecular ions in a standard mixture of benzopyrene, chrysene, carbazole, 7 amino-4-hydroxyquinoline, 5-hydroxyquinoline, 5-aminoquinoline, 5-hydroxy indole, and o-cresol are presented. The results are for a single injection and two passes of the separated material through desorption region at 195/sup 0/C and 285/sup 0/C. Reconstructed ion chromatograms are also presented for eight typical ions in an analysis of a biomass product for a single injection at three different ribbon passes through the flash heater.

  1. RNA footprinting analysis using ion pair reverse phase liquid chromatography.

    PubMed Central

    Dickman, Mark J; Conroy, Matthew J; Grasby, Jane A; Hornby, David P

    2002-01-01

    Hydroxyl radical footprinting is a powerful technique often employed in characterization of the tertiary interactions between proteins and nucleic acids. Following the generation of a nucleic acid "ladder" either by chemical or enzymatic reactions, the radiolabeled products are traditionally separated by denaturing gel electrophoresis and further quantified by phosphorimaging techniques. Here we report the use of ion pair reverse phase liquid chromatography to analyze the products of an RNA footprinting reaction using fluorescently labeled RNA molecules. This technique offers several advantages over existing procedures, including rapid analysis, automation, and direct quantification of the cleavage products without the need to employ radiolabeling. To illustrate the resolving power of this technique, we have analyzed the products of base hydrolysis, generated from a fluorescently labeled RNA molecule and have subsequently used this method to define the solvent accessibility of the substrate strand as it docks with the hairpin ribozyme. PMID:11911369

  2. Ultrapure water for liquid chromatography-mass spectrometry studies.

    PubMed

    Regnault, Cecilia; Kano, Ichiro; Darbouret, Daniel; Mabic, Stéphane

    2004-03-19

    Improvements in trace enrichment techniques combined with the sensitivity of mass spectrometry offer enhanced opportunities to analyze ever lower concentrations of drugs, metabolites, pesticides or environmental pollutants. To perform HPLC and liquid chromatography-mass spectrometry (LC-MS) analyses under optimum conditions, the water used for mobile phase preparation needs to be highly purified and delivered on demand. Indeed, both UV photodiode array detection and MS detection methods are sensitive to organic contaminants (total organic carbon, TOC), and the water quality has a direct impact on the achievable detection limits. The benefits of UV photooxidation on TOC reduction for LC-MS studies were highlighted using electrospray ionization MS detection by comparing HPLC-grade bottled water, freshly produced UV185/254-treated water, and freshly produced non-UV-treated water.

  3. Ultra high pressure liquid chromatography for crude plant extract profiling.

    PubMed

    Eugster, Philippe J; Guillarme, Davy; Rudaz, Serge; Veuthey, Jean-Luc; Carrupt, Pierre-Alain; Wolfender, Jean-Luc

    2011-01-01

    Ultra high pressure liquid chromatography (UHPLC) systems operating at very high pressures and using sub-2 microm packing columns have allowed a remarkable decrease in analysis time and increase in peak capacity, sensitivity, and reproducibility compared to conventional HPLC. This technology has rapidly been widely accepted by the analytical community and is being gradually applied to various fields of plant analysis such as QC, profiling and fingerprinting, dereplication, and metabolomics. For many applications, an important improvement of the overall performances has been reported. In this review, the basic principles of UHPLC are summarized, and practical information on the type of columns used and phase chemistry available is provided. An overview of the latest applications to natural product analysis in complex mixtures is given, and the potential and limitations as well as some new trends in the development of UHPLC are discussed.

  4. Structural analysis of amorphous phosphates using high performance liquid chromatography

    SciTech Connect

    Sales, B.C.; Boatner, L.A.; Chakoumakos, B.C.; McCallum, J.C.; Ramey, J.O.; Zuhr, R.A.

    1993-12-31

    Determining the atomic-scale structure of amorphous solids has proven to be a formidable scientific and technological problem for the past 100 years. The technique of high-performance liquid chromatography (HPLC) provides unique detailed information regarding the structure of partially disordered or amorphous phosphate solids. Applications of the experimental technique of HPLC to phosphate solids are reviewed, and examples of the type of information that can be obtained with HPLC are presented. Inorganic phosphates encompass a large class of important materials whose applications include: catalysts, ion-exchange media, solid electrolytes for batteries, linear and nonlinear optical components, chelating agents, synthetic replacements for bone and teeth, phosphors, detergents, and fertilizers. Phosphate ions also represent a unique link between living systems and the inorganic world.

  5. A rapid liquid chromatography determination of free formaldehyde in cod.

    PubMed

    Storey, Joseph M; Andersen, Wendy C; Heise, Andrea; Turnipseed, Sherri B; Lohne, Jack; Thomas, Terri; Madson, Mark

    2015-01-01

    A rapid method for the determination of free formaldehyde in cod is described. It uses a simple water extraction of formaldehyde which is then derivatised with 2,4-dinitrophenylhydrazine (DNPH) to form a sensitive and specific chromophore for high-performance liquid chromatography (HPLC) detection. Although this formaldehyde derivative has been widely used in past tissue analysis, this paper describes an improved derivatisation procedure. The formation of the DNPH formaldehyde derivative has been shortened to 2 min and a stabilising buffer has been added to the derivative to increase its stability. The average recovery of free formaldehyde in spiked cod was 63% with an RSD of 15% over the range of 25-200 mg kg(-1) (n = 48). The HPLC procedure described here was also compared to a commercial qualitative procedure - a swab test for the determination of free formaldehyde in fish. Several positive samples were compared by both methods.

  6. Determination of sulfonamide residues in eggs by liquid chromatography.

    PubMed

    Furusawa, Naoto

    2002-01-01

    A method was developed for determining residual sulfonamide antibacterials such as sulfamethazine (SMZ), sulfamonomethoxine (SMM), sulfadimethoxine (SDM), and sulfaquinoxaline (SQ) in eggs using liquid chromatography with a photodiode array detector. The spiked and blank samples were cleaned up by using an Ultrafree-MC/PL centrifugal ultrafiltration unit. A Mightysil RP-4 GP column and a mobile phase of 28% (v/v) ethanol-H2O with a photodiode array detector were used for the determination. Average recoveries from eggs spiked with each drug at 0.1, 0.2, 0.4, and 1.0 ppm were > or = 80.9%, with relative standard deviations between 1.3 and 4.7%. The limits of quantitation were 0.060 ppm for SMZ, 0.045 for SMM, 0.044 for SDM, and 0.093 for SQ. The analysis of one sample required < 30 min and < 5 mL ethanol as solvent. PMID:12180677

  7. Nano-liquid chromatography in pharmaceutical and biomedical research.

    PubMed

    Gama, Mariana Roberto; Collins, Carol H; Bottoli, Carla B G

    2013-08-01

    Miniaturized separation techniques have emerged as environmentally friendly alternatives to available separation methods. Nano-liquid chromatography (nano-LC), microchip devices and nano-capillary electrophoresis are miniaturized methods that minimize reagent consumption and waste generation. Furthermore, the low levels of analytes, especially in biological samples, promote the search for more highly sensitive techniques; coupled to mass spectrometry, nano-LC has great potential to become an indispensable tool for routine analysis of biomolecules. This short review presents the fundamental aspects of nano-LC analytical instrumentation, discussing practical considerations and the primary differences between miniaturized and conventional instrumentation. Some theoretical aspects are discussed to better explain both the potential and the principal limitations of nano-LC. Recent pharmaceutical and biomedical applications of this separation technique are also presented to indicate the satisfactory performance for complex matrices, especially for proteomic analysis, that is obtained with nano-LC.

  8. High-pressure liquid chromatography of caffeine in coffee.

    PubMed

    Madison, B L; Kozarek, W J; Damo, C P

    1976-11-01

    A new method is described for the determination of caffeine in coffee, based on high-pressure liquid chromatography. The caffeine is extracted from the sample with water and/or methylene chloride, and then separated from interfering materials by passing an aliquot of the extract through a high-pressure column containing sulfonated cation exchange resin, using 0.01M nitric acid as the mobile phase. An ultraviolet detector measures the absorption of the solution directly. The method is rapid and eliminates the lengthy separations common to other methods. The procedure was applied successfully to decaffeinated and non-decaffeinated green, roasted, and instant coffees. This method gives a more accurate measure of the caffeine content in decaffeinated coffee samples than the micro Bailey-Andrew and modified Levine methods, with equal or better precision. This method gives results equal to those obtained by the official methods for non-decaffeinated samples. PMID:993180

  9. Normal-phase high-performance liquid chromatography of triacylglycerols.

    PubMed

    Rhodes, S H; Netting, A G

    1988-08-31

    Triacylglycerols have been separated by normal-phase high-performance liquid chromatography (HPLC) on silica utilising a solvent system consisting of dry acetonitrile-half water saturated hexane (0.7:99.3). This solvent system is UV transparent allowing detection at 200 nm and affords a separation in which retention is primarily dependent on the number of constituent double bonds. There is also a slight separation on chainlength, the longer chainlengths being eluted first. The system is therefore complementary to currently used reversed-phase HPLC systems. Chromatograms for some polyunsaturated fats and oils are given, and the most polyunsaturated triacylglycerols from linseed oil are analysed in more detail. Data are given for the separation and quantitation of the pentafluorobenzyl esters of constituent fatty acids from these triacylglycerols by a similar normal-phase HPLC system.

  10. Effect of temperature in reversed phase liquid chromatography.

    PubMed

    Guillarme, D; Heinisch, S; Rocca, J L

    2004-10-15

    The high temperature liquid chromatography (HTLC) reveals interesting chromatographic properties but even now, it misses some theoretical aspects concerning the influence of high temperature on thermodynamic and kinetic aspects of chromatography: such a knowledge is very essential for method development. In this work, the effect of temperature on solute behavior has been studied using various stationary phases which are representative of the available thermally stable materials present on the market. The thermodynamic properties were evaluated by using different mobile phases: acetonitrile-water, methanol-water and pure water. The obtained results were discussed on the basis of both type of mobile phases and type of stationary phases. Type of mobile phase was found to play an important role on the retention of solutes. The kinetic aspect was studied at various temperatures ranging from ambient temperature to high temperature (typically from about 30 to 200 degrees C) by fitting the experimental data with the Knox equation and it was shown that the efficiency is improved significantly when the temperature is increased. In this paper, we also discussed the problem of temperature control for thermostating columns which may represent a significant source of peak broadening: by taking into account the three main parameters such as heat transfer, pressure drop and band broadening resulting from the preheating tube, suitable rules are set up for a judicious choice of the column internal diameter. PMID:15527119

  11. Group type analysis of asphalt by column liquid chromatography

    SciTech Connect

    Zhang, C.; Yang, J.; Xue, Y.; Li, Y.

    2008-07-01

    An improved analysis method for characterization of asphalt was established. The method is based on column chromatography technique. The asphalts were separated into four groups: saturates, aromatics, resins, and asphaltenes, quantitatively. About 0.1 g of sample was required in each analysis. About 20 mL of n-heptanes was used to separate out saturates first. Then about 35 mL of n-heptanes/dichloromethane (.5, v/v) mixture was used to separate out aromatics. About 30 mL of dichloromethane/tetrahydrofuran (1/3, v/v) mixture was used to separate out resin. The quality of the separation was confirmed by infrared spectra (IR) and {sup 1}H NMR analysis. The model compounds, tetracosan for saturates, dibenz(o)anthracen for aromatics, and acetanilide for resins were used for verification. The IR and {sup 1}H NMR analysis of the prepared fractions from the column liquid chromatography were in good agreement that of pure reagents.

  12. Towards Chip Scale Liquid Chromatography and High Throughput Immunosensing

    SciTech Connect

    Ni, J.

    2000-09-21

    This work describes several research projects aimed towards developing new instruments and novel methods for high throughput chemical and biological analysis. Approaches are taken in two directions. The first direction takes advantage of well-established semiconductor fabrication techniques and applies them to miniaturize instruments that are workhorses in analytical laboratories. Specifically, the first part of this work focused on the development of micropumps and microvalves for controlled fluid delivery. The mechanism of these micropumps and microvalves relies on the electrochemically-induced surface tension change at a mercury/electrolyte interface. A miniaturized flow injection analysis device was integrated and flow injection analyses were demonstrated. In the second part of this work, microfluidic chips were also designed, fabricated, and tested. Separations of two fluorescent dyes were demonstrated in microfabricated channels, based on an open-tubular liquid chromatography (OT LC) or an electrochemically-modulated liquid chromatography (EMLC) format. A reduction in instrument size can potentially increase analysis speed, and allow exceedingly small amounts of sample to be analyzed under diverse separation conditions. The second direction explores the surface enhanced Raman spectroscopy (SERS) as a signal transduction method for immunoassay analysis. It takes advantage of the improved detection sensitivity as a result of surface enhancement on colloidal gold, the narrow width of Raman band, and the stability of Raman scattering signals to distinguish several different species simultaneously without exploiting spatially-separated addresses on a biochip. By labeling gold nanoparticles with different Raman reporters in conjunction with different detection antibodies, a simultaneous detection of a dual-analyte immunoassay was demonstrated. Using this scheme for quantitative analysis was also studied and preliminary dose-response curves from an immunoassay of a

  13. Analysis of β-blockers in groundwater using large-volume injection coupled-column reversed-phase liquid chromatography with fluorescence detection and liquid chromatography time-of-flight mass spectrometry.

    PubMed

    Galera, María M; Vázquez, Piedad P; Vázquez, María del Mar P; García, María Dolores G; Amate, Carmen F

    2011-08-01

    Atenolol, nadolol, metoprolol, bisoprolol and betaxolol were simultaneously determined in groundwater samples by large-volume injection coupled-column reversed-phase liquid chromatography with fluorescence detection (LVI-LC-LC-FD) and liquid chromatography-time-of-flight mass spectrometry (LC-TOF-MS). The LVI-LC-LC-FD method combines analyte isolation, preconcentration and determination into a single step. Significant reductions in costs for sample pre-treatment (solvent and solid phases for clean up) and method development times are also achieved. Using LC-TOF-MS, accurate mass measurements within 3 ppm error were obtained for all of the β-blockers studied. Empirical formula information can be obtained by this method, allowing the unequivocal identification of the target compounds in the samples. To increase the sensitivity, a solid-phase extraction step with Oasis MCX cartridge was carried out yielding recoveries of 79-114% (n=5) with RSD 2-7% for the LC-TOF-MS method. SPE gives a high purification of β-blockers compared with the existing methods. A 100% methanol wash was allowed for these compounds with no loss of analytes. Limit of quantification was 1-7 ng/L for LVI-LC-LC-FD and 0.25-5 ng/L for LC-TOF-MS. As a result of selective extraction and effective removal of coextractives, no matrix effect was observed in LVI-LC-LC-FD and LC-TOF-MS analyses. The methods were applied to detect and quantify β-blockers in groundwater samples of Almería (Spain).

  14. Determination of sulfonamides in butter samples by ionic liquid magnetic bar liquid-phase microextraction high-performance liquid chromatography.

    PubMed

    Wu, Lijie; Song, Ying; Hu, Mingzhu; Xu, Xu; Zhang, Hanqi; Yu, Aimin; Ma, Qiang; Wang, Ziming

    2015-01-01

    A novel, simple, and environmentally friendly pretreatment method, ionic liquid magnetic bar liquid-phase microextraction, was developed for the determination of sulfonamides in butter samples by high-performance liquid chromatography. The ionic liquid magnetic bar was prepared by inserting a stainless steel wire into the hollow of a hollow fiber and immobilizing ionic liquid in the micropores of the hollow fiber. In the extraction process, the ionic liquid magnetic bars were used to stir the mixture of sample and extraction solvent and enrich the sulfonamides in the mixture. After extraction, the analyte-adsorbed ionic liquid magnetic bars were readily isolated with a magnet from the extraction system. It is notable that the present method was environmentally friendly since water and only several microliters of ionic liquid were used in the whole extraction process. Several parameters affecting the extraction efficiency were investigated and optimized, including the type of ionic liquid, sample-to-extraction solvent ratio, the number of ionic liquid magnetic bars, extraction temperature, extraction time, salt concentration, stirring speed, pH of the extraction solvent, and desorption conditions. The recoveries were in the range of 73.25-103.85 % and the relative standard deviations were lower than 6.84 %. The experiment results indicated that the present method was effective for the extraction of sulfonamides in high-fat content samples.

  15. Dispersive liquid-liquid-liquid microextraction combined with liquid chromatography for the determination of chlorophenoxy acid herbicides in aqueous samples.

    PubMed

    Tsai, Wan-Chun; Huang, Shang-Da

    2009-11-01

    A novel sample preparation method "Dispersive liquid-liquid-liquid microextraction" (DLLLME) was developed in this study. DLLLME was combined with liquid chromatography system to determine chlorophenoxy acid herbicide in aqueous samples. DLLLME is a rapid and environmentally friendly sample pretreatment method. In this study, 25microL of 1,1,2,2-tetrachloroethane was added to the sample solution and the targeted analytes were extracted from the donor phase by manually shaking for 90s. The organic phase was separated from the donor phase by centrifugation and was transferred into an insert. Acceptor phase was added to this insert. The analytes were then back-extracted into the acceptor phase by mixing the organic and acceptor phases by pumping those two solutions with a syringe plunger. After centrifugation, the organic phase was settled and removed with a microsyringe. The acceptor phase was injected into the UPLC system by auto sampler. Fine droplets were formed by shaking and pumping with the syringe plunger in DLLLME. The large interfacial area provided good extraction efficiency and shortened the extraction time needed. Conventional LLLME requires an extraction time of 40-60min; an extraction time of approximately 2min is sufficient with DLLLME. The DLLLME technique shows good linearity (r(2)>or=0.999), good repeatability (RSD: 4.0-12.2% for tap water; 5.7-8.5% for river water) and high sensitivity (LODs: 0.10-0.60microg/L for tap water; 0.11-0.95microg/L for river water).

  16. Determination of household chemicals using gas chromatography and liquid chromatography with tandem mass spectrometry.

    PubMed

    Trenholm, Rebecca A; Vanderford, Brett J; Drewes, Jörg E; Snyder, Shane A

    2008-05-01

    A method has been developed for the determination of 24 household high production volume (HPV) chemicals in municipal wastewater systems using solid-phase extraction (SPE) and analyses using both gas chromatography and liquid chromatography, each with tandem mass spectrometry (GC-MS/MS and LC-MS/MS). Target compounds include pesticides, antioxidants, fragrances, plasticizers, preservatives and personal care products. Method reporting limits ranged from 0.1 to 100 ng/L in water and recoveries for most compounds were between 54 and 112%. Household HPVs were consistently detected in raw sewage entering three full-scale wastewater treatment plants. Compounds such as vanillin, DEET, benzophenone, 3-indolebutyric acid, bisphenol A, triclosan and triclocarban were detected in all wastewater influent and effluent samples, but were significantly lower in the effluent. Many of the remaining compounds were detected in the influent, but below detection in effluent samples. Menthol and phenoxyethanol had the highest observed concentrations in influent samples ranging from 1.5 to 13 microg/L for menthol, and 8.8 to 22 microg/L for phenoxyethanol. MGK-11, methylresorcinol, trifluralin, hexabromododecane, acriflavin and atrazine were not detected in any samples. The method described here detects a broad range of HPV chemicals with great sensitivity and selectivity.

  17. Liquid chromatography-fluorescence and liquid chromatography-mass spectrometry detection of tryptophan degradation products of a recombinant monoclonal antibody.

    PubMed

    Nowak, Christine; Ponniah, Gomathinayagam; Cheng, Guilong; Kita, Adriana; Neill, Alyssa; Kori, Yekaterina; Liu, Hongcheng

    2016-03-01

    Light exposure is one of several conditions used to study the degradation pathways of recombinant monoclonal antibodies. Tryptophan is of particular interest among the 20 amino acids because it is the most photosensitive. Tryptophan degradation forms several products, including an even stronger photosensitizer and several reactive oxygen species. The current study reports a specific peptide mapping procedure to monitor tryptophan degradation. Instead of monitoring peptides using UV 214 nm, fluorescence detection with an excitation wavelength of 295 nm and an emission wavelength of 350 nm was used to enable specific detection of tryptophan-containing peptides. Peaks that decreased in area over time are likely to contain susceptible tryptophan residues. This observation can allow further liquid chromatography-mass spectrometry (LC-MS) analysis to focus only on those peaks to confirm tryptophan degradation products. After confirmation of tryptophan degradation, susceptibility of tryptophan residues can be compared based on the peak area decrease. PMID:26717898

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

    DOEpatents

    Johnson, Charles C.; Taylor, Larry T.

    1986-01-01

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

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

    DOEpatents

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

    1985-01-04

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

  20. High Performance Liquid Chromatography at -196 °C.

    PubMed

    Motono, Tomohiro; Kitagawa, Shinya; Ohtani, Hajime

    2016-07-01

    Ultralow temperature high-performance liquid chromatography (HPLC) was developed using a liquefied gas as the mobile phase. HPLC separation of low molecular weight alkanes at -196 °C with liquid nitrogen mobile phase was successfully achieved, whereas their GC separation at -196 °C using helium gas mobile phase failed to elute the analytes due to strong adsorption. Prior to the further study of HPLC at -196 °C, the effect of column temperature on the chromatographic behavior was investigated, and it was found that the retention of analytes drastically increased when the column temperature was over the boiling point of the mobile phase. As the study of retention control in HPLC at -196 °C, the mobile phases of nitrogen and methane mixtures were investigated. The addition of methane to the nitrogen mobile phase suppressed the retention of the analytes (tetra-deuterated methane, ethane, and propane), that is, the retention on HPLC at ultralow temperature could be controlled by the mobile phase composition, akin to the typical retention in HPLC. The selectivity toward the n- and iso-alkane in HPLC at -196 °C was altered compared with that in GC separation at room temperature. A significant enhancement of retention of alkanes compared with alkanes were observed in HPLC at -196 °C. PMID:27282809

  1. Determination of doxycycline in chicken fat by liquid chromatography with UV detection and liquid chromatography-tandem mass spectrometry.

    PubMed

    Gajda, Anna; Posyniak, Andrzej; Zmudzki, Jan; Tomczyk, Grzegorz

    2013-06-01

    A sensitive analytical method for determination of doxycycline (DC) residues in chicken fat/fat and skin was developed. The extraction, in the presence of the internal standard (IS) minocycline (MINO), was carried out using solution of oxalic acid (pH 4.0) and ethyl acetate. The samples were cleaned up by solid phase extraction (SPE) procedure using, at first carboxylic acid and then polymeric Strata X cartridges. Chromatographic separation of DC by LC-UV was achieved on a Luna C8 analytical column and for LC-MS/MS analysis Luna C18 column was used. The presented procedures were evaluated according to the Commission Decision 2002/657/EC. Specificity, decision limit (CCα), detection capacity (CCβ), recovery (absolute and relative), precision (repeatability and reproducibility) were determined during validation process. The limit of detection (LOD) was 10μg/kg for LC-UV and 1μg/kg for LC-MS/MS method. The limit of quantitation (LOQ) was 15 and 2μg/kg for LC-UV and LC-MS/MS, respectively. The absolute recovery for the LC-UV and relative recovery for the LC-MS/MS method at 300μg/kg concentration level were 79%; 101% for fat and 82%; 99% for fat and skin, respectively. The developed liquid chromatography with ultraviolet detection (LC-UV) and liquid chromatography tandem mass spectrometry (LC-MS/MS) methods have been applied to quantitative determination of doxycycline (DC) in samples of chicken fat tissue obtained from animals treated with DC.

  2. Percutaneous Cystgastrostomy as a Single-Step Procedure

    SciTech Connect

    Curry, L. Sookur, P.; Low, D.; Bhattacharya, S.; Fotheringham, T.

    2009-03-15

    The purpose of this study was to evaluate the success of percutaneous transgastric cystgastrostomy as a single-step procedure. We performed a retrospective analysis of single-step percutaneous transgastric cystgastrostomy carried out in 12 patients (8 male, 4 female; mean age 44 years; range 21-70 years), between 2002 and 2007, with large symptomatic pancreatic pseudocysts for whom up to 1-year follow-up data (mean 10 months) were available. All pseudocysts were drained by single-step percutaneous cystgastrostomy with the placement of either one or two stents. The procedure was completed successfully in all 12 patients. The pseudocysts showed complete resolution on further imaging in 7 of 12 patients with either enteric passage of the stent or stent removal by endoscopy. In 2 of 12 patients, the pseudocysts showed complete resolution on imaging, with the stents still noted in situ. In 2 of 12 patients, the pseudocysts became infected after 1 month and required surgical intervention. In 1 of 12 patients, the pseudocyst showed partial resolution on imaging, but subsequently reaccumulated and later required external drainage. In our experience, percutaneous cystgastrostomy as a single-step procedure has a high success rate and good short-term outcomes over 1-year follow-up and should be considered in the treatment of large symptomatic cysts.

  3. Methods to approximate reliabilities in single-step genomic evaluation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Reliability of predictions from single-step genomic BLUP (ssGBLUP) can be calculated by inversion, but that is not feasible for large data sets. Two methods of approximating reliability were developed based on decomposition of a function of reliability into contributions from records, pedigrees, and...

  4. Determination of descriptors for fragrance compounds by gas chromatography and liquid-liquid partition.

    PubMed

    Karunasekara, Thushara; Poole, Colin F

    2012-04-27

    Retention factors on a minimum of eight stationary phases at various temperatures by gas-liquid chromatography and liquid-liquid partition coefficients for five totally organic biphasic systems were combined to estimate descriptors for 28 fragrance compounds with an emphasis on compounds that are known or potential allergens. The descriptors facilitated the estimation of several properties of biological and environmental interest (sensory irritation threshold, odor detection threshold, nasal pungency threshold, skin permeability from water, skin-water partition coefficients, octanol-water partition coefficients, absorption by air particles, adsorption by diesel soot particles, air-water partition coefficients, and adsorption by film water). The descriptors are suitable for use in the solvation parameter model and facilitate the estimation of a wide range of physicochemical, chromatographic, biological, and environmental properties using existing models.

  5. Identification of New Metabolites of Bacterial Transformation of Indole by Gas Chromatography-Mass Spectrometry and High Performance Liquid Chromatography

    PubMed Central

    Arora, Pankaj Kumar

    2014-01-01

    Arthrobacter sp. SPG transformed indole completely in the presence of an additional carbon source. High performance liquid chromatography and gas chromatography-mass spectrometry detected indole-3-acetic acid, indole-3-glyoxylic acid, and indole-3-aldehyde as biotransformation products. This is the first report of the formation of indole-3-acetic acid, indole-3-glyoxylic acid, and indole-3-aldehyde from indole by any bacterium. PMID:25548566

  6. Determination of L-ephedrine, pseudoephedrine, and caffeine in rat plasma by liquid chromatography-tandem mass spectrometry.

    PubMed

    Cooper, Stephen D; Fletcher, Brenda L; Silinski, Melanie A Rehder; Brown, Sherri S; Lodge, Jon W; Fernando, Reshan A; Collins, Bradley J

    2011-07-01

    A rapid and simple liquid chromatography tandem mass spectrometry method was developed and validated for the simultaneous determination of L-ephedrine, pseudoephedrine, and caffeine in male Fisher-344 rat plasma at nanogram-per-milliliter concentrations for use in support of toxicology studies. Only 25 μL of plasma is required, and extraction is performed using a simple, single-step protein precipitation. The method was validated over a range of 2.09 to 5460 ng/mL for L-ephedrine, 2.09 to 5050 ng/mL for pseudoephedrine and 2.03 to 5340 ng/mL for caffeine. A binary gradient elution at 0.3 mL/min was used with a Waters XBridge Phenyl (2.1 × 150 mm, 3.5 μm) column and a Waters XBridge Phenyl 2.1- × 10-mm guard column at ambient temperature. The mobile phase consisted of 10 mM ammonium acetate in water (pH 5.0) and methanol. Caffeine trimethyl-(13)C(3) was used as the internal standard. The method was evaluated for linearity, recovery, precision, accuracy, and stability, and it was successfully applied in toxicokinetic studies of ephedrine, administered alone, in combination with caffeine, and in the herbal source Ma Huang.

  7. [Advances in multidimensional high performance liquid chromatography for separation technology in proteomic study].

    PubMed

    Gao, Mingxia; Guan, Xia; Hong, Guangfeng; Zhang, Xiangmin

    2009-09-01

    With the developments of proteomic study, multidimensional high performance liquid chromatography technology has attracted increasing interests due to its obvious advantages, such as rapid analysis, high automation and easy combination with mass spectrometry and so on. This review emphasizes the advances of multidimensional high performance liquid chromatography technology, including classical bottom-up methods, top-down techniques and array-based two-dimensional liquid chromatography system, which was designed and set up by our lab to improve the throughput. These techniques showed promising potential applications in proteomics study.

  8. Simultaneous determination of cyclodol and diprazin by thin layer chromatography and high performance liquid chromatography.

    PubMed

    Makharadze, R; Adeishvili, L; Chelidze, T; Imnadze, N; Nizharadze, N

    2009-11-01

    Ciklodol (trihexyphenidil)--the central and peripheral m-cholinoblocker is currently used with other antipsychotic drugs such as phenotiazines and tricycle antidepressants. For the purpose of simultaneous determination of ciklodol and diprazine, were selected two methods of analysis: Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC). During development of TLC method was studied the 10 visualizing system and 24 mobile systems. For individual or simultaneous determination of ciklodol and diprazine were recommended the following solvents' systems: 1. Toluene-acetone-ethanole-25%NH(4)OH (45:45: 7.5:2.5), 2. Hexane-ethyl acetate (15:5), 3. Chloroform-heptene-25%NH(4)OH (16:3:3), 4. Ethylacetate-hexane (10:10), 5. Acetonitrile-metanol (10:10) and 6.Heptene-chloroform-ethanol-25% NH(4)OH (5:10:3:1). As visualizing systems were chosen: Iodine vapors, blacklight (UV254) and reagent of FNP. Reagent of FNP gives colored spot just with diprazine and it is also could be used for separation of both objects in simultaneous analysis. Developed HPLC method of simultaneous determination of ciklodol and diprazine: like mobile phase is recommended: Acetonitril- 0.05M KH(2)PO4 (55:45) (v/v) +H(3)PO(4) (pH3.5), column EC250 x 4.6mm, with solid phase Nucleosil, flow rate 1ml/min, sample volume 40 microl. In given conditions, the retention time of ciklodol is 6.005min and diprazine 7.227min. Developed method of simultaneous determination and separation of ciklodol and diprazine in respective mixtures could be successfully applied as in the pharmaceutical, as well in the chemical-toxicological laboratories.

  9. Unified pH values of liquid chromatography mobile phases.

    PubMed

    Suu, Agnes; Jalukse, Lauri; Liigand, Jaanus; Kruve, Anneli; Himmel, Daniel; Krossing, Ingo; Rosés, Martí; Leito, Ivo

    2015-03-01

    This work introduces a conceptually new approach of measuring pH of mixed-solvent liquid chromatography (LC) mobile phases. Mobile phase pH is very important in LC, but its correct measurement is not straightforward, and all commonly used approaches have deficiencies. The new approach is based on the recently introduced unified pH (pH(abs)) scale, which enables direct comparison of acidities of solutions made in different solvents based on chemical potential of the proton in the solutions. This work represents the first experimental realization of the pH(abs) concept using differential potentiometric measurement for comparison of the chemical potentials of the proton in different solutions (connected by a salt bridge), together with earlier published reference points for obtaining the pH(abs) values (referenced to the gas phase) or pH(abs)(H₂O) values (referenced to the aqueous solution). The liquid junction potentials were estimated in the framework of Izutsu's three-component method. pH(abs) values for a number of common LC and LC-MS mobile phases have been determined. The pH(abs) scale enables for the first time direct comparison of acidities of any LC mobile phases, with different organic additives, different buffer components, etc. A possible experimental protocol of putting this new approach into chromatographic practice has been envisaged and its applicability tested. It has been demonstrated that the ionization behavior of bases (cationic acids) in the mobile phases can be better predicted by using the pH(abs)(H₂O) values and aqueous pKa values than by using the alternative means of expressing mobile phase acidity. Description of the ionization behavior of acids on the basis of pH(abs)(H₂O) values is possible if the change of their pKa values with solvent composition change is taken into account. PMID:25664372

  10. Isopropylammonium Formate as a Mobile Phase Modifier for Liquid Chromatography

    PubMed Central

    Collins, Matthew P.; Zhou, Ling; Camp, Suzanne E.; Danielson, Neil D.

    2012-01-01

    Isopropylammonium formate (IPAF), a new alkylammonium formate (AAF) room temperature ionic liquid, has been synthesized from isopropylamine and formic acid and characterized as an organic solvent mobile phase replacement for reversed-phase liquid chromatography (LC). Characterization of IPAF solvent properties in water such as pH, conductivity, and viscosity, as well as its synthesis, is described. The LC polarity (P′) and the solvent strength (S) parameters are determined to be 6.0 and 2.4, respectively, similar to those same parameters for methanol and acetonitrile. Application of this RTIL is demonstrated as an organic solvent replacement for reversed-phase LC to separate a test mixture of niacinamide, acetophenone and p-nitroaniline. The van Deemter plot profile for several columns of different dimensions, particle size, pore size and stationary phase are compared using an IPAF–water mobile phase. At flow rates above 2 mL/min, on-line mixing of the viscous IPAF with water appears not to be uniform. A flattening of the van Deemter profile is noted for particularly short (50 mm) wide bore (4.6 mm) columns packed with larger particles (10 µm). Small particle longer columns likely facilitated mixing at the beginning of the column generating typical linearly increasing van Deemeter curves. IPAF has been further shown as a function of temperature to be a non-denaturing modifier solvent for the separation of the protein cytochrome c from tryptophan compared to methanol. This is important to show, because the semi-preparative separation of native proteins using AAF mobile phases is the long-term goal of this research program. PMID:22718743

  11. Simultaneous analysis for water- and fat-soluble vitamins by a novel single chromatography technique unifying supercritical fluid chromatography and liquid chromatography.

    PubMed

    Taguchi, Kaori; Fukusaki, Eiichiro; Bamba, Takeshi

    2014-10-01

    Chromatography techniques usually use a single state in the mobile phase, such as liquid, gas, or supercritical fluid. Chromatographers manage one of these techniques for their purpose but are sometimes required to use multiple methods, or even worse, multiple techniques when the target compounds have a wide range of chemical properties. To overcome this challenge, we developed a single method covering a diverse compound range by means of a "unified" chromatography which completely bridges supercritical fluid chromatography and liquid chromatography. In our method, the phase state was continuously changed in the following order; supercritical, subcritical and liquid. Moreover, the gradient of the mobile phase starting at almost 100% CO2 was replaced with 100% methanol at the end completely. As a result, this approach achieved further extension of the polarity range of the mobile phase in a single run, and successfully enabled the simultaneous analysis of fat- and water-soluble vitamins with a wide logP range of -2.11 to 10.12. Furthermore, the 17 vitamins were exceptionally separated in 4min. Our results indicated that the use of dense CO2 and the replacement of CO2 by methanol are practical approaches in unified chromatography covering diverse compounds. Additionally, this is a first report to apply the novel approach to unified chromatography, and can open another door for diverse compound analysis in a single chromatographic technique with single injection, single column and single system.

  12. Single-step route to diamond-nanotube composite

    PubMed Central

    2012-01-01

    Candle wax was used as a precursor for the production of a diamond-nanotube composite in a single step. The composite films were fabricated by sulfur-assisted hot-filament chemical vapor deposition technique. The morphology of the composite films was analyzed by scanning electron microscopy and transmission electron microscopy. Raman spectra of the films show characteristic diamond band at 1,332 cm−1, D-band around 1,342 cm−1, and graphitic G-band around 1,582 cm−1. The electron energy-loss spectroscopy recorded at the carbon K-edge region shows signature features of diamond and carbon nanotube in the fabricated material. The ability to synthesize diamond-nanotube composites at relatively low temperatures by a single-step process opens up new possibilities for the fabrication of nanoelectronic devices. PMID:23013660

  13. Single-step route to diamond-nanotube composite

    NASA Astrophysics Data System (ADS)

    Varshney, Deepak; Ahmadi, Majid; Guinel, Maxime J.-F.; Weiner, Brad R.; Morell, Gerardo

    2012-09-01

    Candle wax was used as a precursor for the production of a diamond-nanotube composite in a single step. The composite films were fabricated by sulfur-assisted hot-filament chemical vapor deposition technique. The morphology of the composite films was analyzed by scanning electron microscopy and transmission electron microscopy. Raman spectra of the films show characteristic diamond band at 1,332 cm-1, D-band around 1,342 cm-1, and graphitic G-band around 1,582 cm-1. The electron energy-loss spectroscopy recorded at the carbon K-edge region shows signature features of diamond and carbon nanotube in the fabricated material. The ability to synthesize diamond-nanotube composites at relatively low temperatures by a single-step process opens up new possibilities for the fabrication of nanoelectronic devices.

  14. Liquid chromatography of hydrocarbonaeous quaternary amines on cyclodextrin bonded silica

    USGS Publications Warehouse

    Abidi, S.L.

    1986-01-01

    Mixtures of n-alkylbenzyldimethylammonium chloride (ABDAC) were resolved into homologous components by high-performance liquid chromatography (HPLC) with a cyclodextrin-bonded silica stationary phase. With a few exceptions, results from this study are similar to those obtained from traditional reversed-phase HPLC. It was found that the presence of electrolytes in aqueous mobile phases is not a critical factor in determining the success of HPLC separation. Under normal HPLC conditions, a mobile phase consisting of either methanol–water (50:50) or acetonitrile–water (30:70) was employed for obtaining adequate resolution of the quaternary ammonium mixtures. Although the percent organic modifier–water profiles were similar to those in previous studies with these compounds, resolution (R) and selectivity (α) parameters were found to be quite susceptible to changes in the mobile phase solvent composition. The retention behavior of the cationic analytes in the homologous series is consistent with the hydrophobic-interaction concept proposed for the retention mechanism via dominant inclusion complex formation. Several electrolytes were chosen for a study of the counter ion effect on the chromatographic characteristics of ABDAC components. Among the electrolytes examined, the perchlorate ion was found most likely to act as an ion-pairing counter ion for ammonium cations in the HPLC system studied. A correlation study established linear relationships between the chain length of ABDAC and the logarithmic capacity factor (k2). The analytical utility of the HPLC method was demonstrated by the analysis of various unknown mixtures.

  15. Simultaneous determination of sulfaquinoxaline, sulfamethazine and pyrimethamine by liquid chromatography.

    PubMed

    Berzas Nevado, J J; Castañeda Peñalvo, G; Guzmán Bernardo, F J

    2000-02-18

    A liquid chromatography method is described to determine sulfaquinoxaline (SQX), sulfamethazine (SMT), and pyrimethamine (PMT), by using a Kromasil C18 column and a 40 mM NaH2PO4 buffer solution, containing 10 mM NaClO4 (pH 3.0)-acetonitrile (65:35) as mobile phase. The mobile phase flow-rate and sample volume injected were 1.5 ml/min and 20 microl, respectively and the samples were dissolved in the mobile phase. The limits of quantification were found to be about 180 microg/l (3.6 ng) for each compound. The method was applied in veterinary commercial formulations. Analyses were made by means of the standard addition method, whose results were compared with those obtained by preparing "tests" (from the stock solutions) and with those obtained by a capillary electrophoresis method. Both methods showed similar results, and then it was proved that some commercial claimed levels were not in agreement with the obtained results by using our analytical method, as they were in other cases. PMID:10722074

  16. Chromatographic analysis of olopatadine in hydrophilic interaction liquid chromatography.

    PubMed

    Maksić, Jelena; Jovanović, Marko; Rakić, Tijana; Popović, Igor; Ivanović, Darko; Jančić-Stojanović, Biljana

    2015-01-01

    In this paper, chromatographic analysis of active substance olopatadine hydrochloride, which is used in eye drops as antihistaminic agent, and its impurity E isomer by hydrophilic interaction liquid chromatography (HILIC) and application of design of experiments (DoE) methodology are presented. In addition, benzalkonium chloride is very often used as a preservative in eye drops. Therefore, the evaluation of its chromatographic behavior in HILIC was carried out as well. In order to estimate chromatographic behavior and set optimal chromatographic conditions, DoE methodology was applied. After the selection of important chromatographic factors, Box-Behnken design was utilized, and on the basis of the obtained models factor effects were examined. Then, multi-objective robust optimization is performed aiming to obtain chromatographic conditions that comply with several quality criteria simultaneously: adequate and robust separation of critical peak pair and maximum retention of the first eluting peak. The optimal conditions are identified by using grid point search methodology. The experimental verification confirmed the adequacy of the defined optimal conditions. Finally, under optimal chromatographic conditions, the method was validated and applicability of the proposed method was confirmed.

  17. Determination of saffron quality by high-performance liquid chromatography.

    PubMed

    Valle García-Rodríguez, M; Serrano-Díaz, Jéssica; Tarantilis, Petros A; López-Córcoles, Horacio; Carmona, Manuel; Alonso, Gonzalo L

    2014-08-13

    The aim of this work was to propose a high-performance liquid chromatography with diode array detection (HPLC-DAD) method for determining the three main compounds responsible for determining the quality of saffron (crocetin esters, picrocrocin, and safranal) by preparing an aqueous extract according to the ISO 3632 standard to solve the difficulty that this standard has for aroma and taste determination by ultraviolet-visible spectroscopy. Toward this aim, laboratory-isolated picrocrocin, a safranal standard with a purity of ≥ 88%, trans-crocetin di(β-D-gentiobiosyl) ester (trans-4-GG) and trans-crocetin (β-D-glucosyl)-(β-D-gentiobiosyl) ester (trans-3-Gg) standards, both with a purity of ≥ 99%, and 50 different saffron spice samples from Italy, Iran, Greece, and Spain were used in the intralaboratory validation of the HPLC method. The analytical method proposed was adequate in terms of linearity, selectivity, sensitivity, and accuracy for determining the three foremost parameters that define the quality of saffron using only a saffron solution prepared according to the ISO 3632 standard.

  18. Electrochemically modulated liquid chromatography: Theoretical investigations and applications from the perspectives of chromatography and interfacial electrochemistry

    SciTech Connect

    Keller, David W.

    2005-01-01

    Electrochemically modulated liquid chromatography (EMLC) employs a conductive material as both a stationary phase for chromatographic separations and as a working electrode for performing electrochemistry experiments. This dual functionality gives EMLC the capacity to manipulate chromatographic separations by changing the potential applied (Eapp) to the stationary phase with respect to an external reference. The ability to monitor retention as a function of Eapp provides a means to chromatographically monitor electrosorption processes at solid-liquid interfaces. In this dissertation, the retention mechanism for EMLC is examined from the perspective of electrical double layer theory and interfacial thermodynamics. From the chromatographic data, it is possible to determine the interfacial excess (Λ) of a solute and changes in interfacial tension (dγ) as a function of both Eapp and the supporting electrolyte concentration. Taken together, these two experimentally manipulated parameters can be examined within the context of the Gibbs adsorption equation to delineate the contribution of a variety of interfacial properties, including the charge of solute on the stationary phase and the potential of zero charge (PZC), to the mechanism behind EMLC-based retention. The chromatographic probing of interfacial phenomena is complemented by electroanalytical experiments that exploit the ability to monitor the electronic current flowing through an EMLC column. Cyclic voltammetry and chronoamperometry of an EMLC column are used to determine the electronic performance characteristics of an EMLC column. An electrochemical flow injection analysis of a column is provided in which the current required to maintain a constant Eapp is monitored and provides a way to examine the influence that acetonitrile and supporting electrolyte composition, flow rate, column backpressure, and ionic strength have on the structure of electrified interfaces.

  19. Preparation of fatty acid methyl esters for gas-liquid chromatography[S

    PubMed Central

    Ichihara, Ken'ichi; Fukubayashi, Yumeto

    2010-01-01

    A convenient method using commercial aqueous concentrated HCl (conc. HCl; 35%, w/w) as an acid catalyst was developed for preparation of fatty acid methyl esters (FAMEs) from sterol esters, triacylglycerols, phospholipids, and FFAs for gas-liquid chromatography (GC). An 8% (w/v) solution of HCl in methanol/water (85:15, v/v) was prepared by diluting 9.7 ml of conc. HCl with 41.5 ml of methanol. Toluene (0.2 ml), methanol (1.5 ml), and the 8% HCl solution (0.3 ml) were added sequentially to the lipid sample. The final HCl concentration was 1.2% (w/v). This solution (2 ml) was incubated at 45°C overnight or heated at 100°C for 1–1.5 h. The amount of FFA formed in the presence of water derived from conc. HCl was estimated to be <1.4%. The yields of FAMEs were >96% for the above lipid classes and were the same as or better than those obtained by saponification/methylation or by acid-catalyzed methanolysis/methylation using commercial anhydrous HCl/methanol. The method developed here could be successfully applied to fatty acid analysis of various lipid samples, including fish oils, vegetable oils, and blood lipids by GC. PMID:19759389

  20. Determination of Aspartame, Caffeine, Saccharin, and Benzoic Acid in Beverages by High Performance Liquid Chromatography.

    ERIC Educational Resources Information Center

    Delaney, Michael F.; And Others

    1985-01-01

    Describes a simple and reliable new quantitative analysis experiment using liquid chromatography for the determinaiton of caffeine, saccharin, and sodium benzoate in beverages. Background information, procedures used, and typical results obtained are provided. (JN)

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

    NASA Technical Reports Server (NTRS)

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

    1976-01-01

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

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

    SciTech Connect

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

    1986-07-10

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

  3. HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF SELECTED ORGANIC PEROXIDES WITH OXIDATIVE AMPEROMETRIC DETECTION

    EPA Science Inventory

    Reversed-phase high-performance liquid chromatography with oxidative amperometric detection was optimized for the determination of several organic peroxides in drinking water under ideal conditions.The determinations were performed under isocratic conditions using acetonitrile an...

  4. New methods and materials for solid phase extraction and high performance liquid chromatography

    SciTech Connect

    Dumont, P.J.

    1996-04-23

    This paper describes methods for solid phase extraction and high performance liquid chromatography (HPLC). The following are described: Effects of Resin Sulfonation on the Retention of Polar Organic Compounds in Solid Phase Extraction; Ion-Chromatographic Separation of Alkali Metals In Non-Aqueous Solvents; Cation-Exchange Chromatography in Non-Aqueous Solvents; and Silicalite As a Stationary Phase For HPLC.

  5. A simple liquid extraction protocol for overcoming the ion suppression of triacylglycerols by phospholipids in liquid chromatography mass spectrometry studies.

    PubMed

    Araujo, Pedro; Tilahun, Ephrem; Breivik, Joar Fjørtoft; Abdulkader, Bashir M; Frøyland, Livar; Zeng, Yingxu

    2016-02-01

    It is well-known that triacylglycerol (TAG) ions are suppressed by phospholipid (PL) ions in regiospecific analysis of TAG by mass spectrometry (MS). Hence, it is essential to remove the PL during sample preparation prior to MS analysis. The present article proposes a cost-effective liquid-liquid extraction (LLE) method to remove PL from TAG in different kinds of biological samples by using methanol, hexane and water. High performance thin layer chromatography confirmed the lack of PL in krill oil and salmon liver samples, submitted to the proposed LLE protocol, and liquid chromatography tandem MS confirmed that the identified TAG ions were highly enhanced after implementing the LLE procedure.

  6. Column liquid chromatography-ultraviolet and column liquid chromatography/mass spectrometry evaluation of stress degradation behavior of escitalopram oxalate.

    PubMed

    Dhaneshwar, Sunil R; Mahadik, Mahadeo V; Kulkarni, Mahesh J

    2009-01-01

    The objective of this work was to study the degradation behavior of escitalopram oxalate under different International Conference on Harmonization (ICH)-recommended stress conditions by column liquid chromatography (LC)-UV and LC/mass spectrometry (LC/MS) and to establish a validated stability-indicating LC assay method. Escitalopram oxalate was subjected to stress conditions of hydrolysis, oxidation, photolysis, and thermal decomposition. Extensive degradation was found to occur in alkaline medium. Mild degradation was observed in acidic and oxidative conditions. Escitalopram oxalate was stable to neutral, photolytic, and thermal stress. Successful separation of the drug from degradation products formed under stress conditions was achieved on a PerfectSil-100 ODS-3 column [C18 (5 microm, 25 cm x 4.6 mm id)] using methanol-0.01 M acetate buffer pH 3.8 adjusted with acetic acid (45 + 55) as the mobile phase. The flow rate was 1 ml/min, and the detection wavelength was 239 nm. The method was validated according to ICH guidelines. Major degradation products formed in hydrolysis and oxidative conditions were isolated, and structural elucidation of degradation products was done by LCIMS and infrared spectrometry studies. The major hydrolysis degradation product was confirmed as 1-(3-dimethylaminopropyl)-1-(4-fluoro- phenyl)-1,3dihydroisobenzofuran-5-carboxylic acid, and the major oxidative degradation product was confirmed as 1-{[3-dimethylamino(oxide)- propyl]-1-(4-fluro-phenyl)}-1,3-dihydro-isobenzofuran- 5-carbonitrile.

  7. Chromatography.

    ERIC Educational Resources Information Center

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

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

  8. Advanced Capillary Liquid Chromatography-Mass Spectrometry for Proteomics

    SciTech Connect

    Shen, Yufeng; Page, Jason S.; Smith, Richard D.

    2009-02-23

    The liquid chromatography (LC)-mass spectrometric (MS) analysis of peptides has become a routine method for proteomics – the study of the entire complement of proteins e.g., expressed by a cell under a specific set of conditions at a specific time. Mixtures of peptides, such as those generated from enzymatic (e.g., trypsin) digestion of globally recovered proteins (i.e. a proteome), are typically very complex and >100,000 different molecular species may be observable using MS detection [1]. LC separations implemented prior to MS for broad protein identification have three major roles: 1) to isolate individual components or reduce complexity as much as possible, 2) to increase sensitivity by concentrating the components into narrow zones prior to MS, and 3) to eliminate or displace interfering species (e.g., salts and polymers) that may be present in proteomics samples. A desired quality of LC separation can be achieved from the use of either multiple steps of moderate quality separations, or fewer steps of high power separations. The former approach is generally more easily accessible for very high quality separations due to the variety of commercialized LC platforms available, while the latter still often requires considerable developmental efforts (for both columns and instrumentation). In addition to proteomics data quality, other differences between these two approaches include proteomics analysis time and sample consumption (and subsequent analysis costs), as well as direct impact on potential proteomics applications that have special requirements in terms of analysis coverage, sample size, dynamic range, sensitivity, and throughput.

  9. Determination of dalcetrapib by liquid chromatography-tandem mass spectrometry.

    PubMed

    Heinig, Katja; Bucheli, Franz; Kuhlmann, Olaf; Zell, Manfred; Pähler, Axel; Zwanziger, Elke; Gross, Günter; Tardio, Joseph; Ishikawa, Tomohiro; Yamashita, Tomoko

    2012-07-01

    The cholesteryl ester transfer protein modulator dalcetrapib is currently under development for the prevention of dyslipidemia and cardiovascular disease. Dalcetrapib, a thioester, is rapidly hydrolyzed in vivo to the corresponding thiophenol which in turn is further oxidized to the dimer and mixed disulfides (where the thiophenol binds to peptides, proteins and other endogenous thiols). These forms co-exist in an oxidation-reduction equilibrium via the thiol and cannot be stabilized without influencing the equilibrium, hence specific determination of individual components, i.e., in order to distinguish between the free thiol, the disulfide dimer and mixed disulfide adducts, was not pursued for routine analysis. The individual forms were quantified collectively as dalcetrapib-thiol (dal-thiol) after reduction under basic conditions with dithiothreitol to break disulfide bonds and derivatization with N-ethylmaleimide to stabilize the free thiol. The S-methyl and S-glucuronide metabolites were determined simultaneously with dal-thiol with no effect from the derivatization procedure. Column-switching liquid chromatography-tandem mass spectrometry provided a simple, fast and robust method for analysis of human and animal plasma and human urine samples. Addition of the surfactant Tween 80 to urine prevented adsorptive compound loss. The lower limits of quantitation (LLOQ) were 5 ng/mL for dal-thiol, and 5 ng/mL for the S-methyl and 50 ng/mL for the S-glucuronide metabolites. Using stable isotope-labeled internal standards, inter- and intra-assay precisions were each <15% (<20% at LLOQ) and accuracy was between 85 and 115%. Recovery was close to 100%, and no significant matrix effect was observed. PMID:22541249

  10. Using dispersive liquid-liquid microextraction and liquid chromatography for determination of guaifenesin enantiomers in human urine.

    PubMed

    Hatami, Mehdi; Farhadi, Khalil; Abdollahpour, Assem

    2011-11-01

    A simple, rapid, and efficient method, dispersive liquid-liquid microextraction (DLLME) coupled with high-performance liquid chromatography-fluorescence detector, has been developed for the determination of guaifenesin (GUA) enantiomers in human urine samples after an oral dose administration of its syrup formulation. Urine samples were collected during the time intervals 0-2, 2-4, and 4-6 h and concentration and ratio of two enantiomers was determined. The ratio of R-(-) to S-(+) enantiomer concentrations in urine showed an increase with time, with R/S ratios of 0.66 at 2 h and 2.23 at 6 h. For microextraction process, a mixture of extraction solvent (dichloromethane, 100 μL) and dispersive solvent (THF, 1 mL) was rapidly injected into 5.0 mL diluted urine sample for the formation of cloudy solution and extraction of enantiomers into the fine droplets of CH(2)Cl(2). After optimization of HPLC enantioselective conditions, some important parameters, such as the kind and volume of extraction and dispersive solvents, extraction time, temperature, pH, and salt effect were optimized for dispersive liquid-liquid microextraction process. Under the optimum extraction condition, the method yields a linear calibration curve in the concentration range from 10 to 2000 ng/mL for target analytes. LOD was 3.00 ng/mL for both of the enantiomers. PMID:21972192

  11. Vortex and air assisted liquid-liquid microextraction as a sample preparation method for high-performed liquid chromatography determinations.

    PubMed

    Hosseini, Mohammad; Heydari, Rouhollah; Alimoradi, Mohammad

    2014-12-01

    A novel, simple and sensitive method based on vortex and air assisted liquid-liquid microextraction (VAALLME) technique coupled with high-performance liquid chromatography (HPLC) has been developed for quantitative analysis of β-naphthol, naphthalene and anthracene as model analytes. Unlike the dispersive liquid-liquid microextraction (DLLME), dispersive solvent and centrifuging step were eliminated in proposed technique. In this technique, extraction solvent was dispersed into the aqueous sample solution by using vortex. Phase separation was achieved via motion of air bubbles from the bottom to top of the extraction tube, which promoted the analytes transfer into the supernatant organic phase. Influential parameters on the extraction efficiency such as type and volume of extraction solvent, salt type and its concentration, vortex and aeration times, and sample pH were evaluated and optimized. The calibration curves showed good linearity (r(2)>0.9947) and precision (RSD<5.0%) in the working concentration ranges. The limit of detection (LOD) for β-naphthol, naphthalene and anthracene were 10, 5.0 and 0.5 ng mL(-1), respectively. The recoveries were in the range of 97.0-102.0% with RSD values ranging from 2.2 to 5.2%.

  12. [Determination of phenolic and salicylanilide anthelmintics in liquid milk by high performance liquid chromatography].

    PubMed

    Wang, Wei; Huang, Xianhui; Wang, Hui; Yan, Changyan; Kong, Xiangkai

    2013-10-01

    An analytical method for the determination of nitroxynil, oxyclozanide, closantel and rafoxanide in liquid milk by high performance liquid chromatography (HPLC) has been established. The milk sample was extracted with acetonitrile containing 1% (v/v) triethylamine. The supernatant was purified by an anion exchange solid phase extraction column. The analyte was detected by ultraviolet detector after the HPLC separation on a C18 RP column. The mobile phase was composed of acetonitrile-0.02 mol/L ammonium acetate solution with pH 4.0. The linear ranges of the four drugs in the spiked blank milk samples were 5-500 microg/kg, and the correlation coefficients were higher than 0.99. The limits of detection (LOD) were 3 microg/kg, and the limits of quantification (LOQ) were 5 microg/kg. The average recoveries and relative standard deviations (RSDs) of nitroxynil, oxyclozanide, closantel and rafoxanide at the spiked levels of 1/2MRL (maximum residue limit), MRL and 2MRL ranged from 92.20% to 96.13%, 5.55% to 16.30%; 87.40% to 94.74%, 5.40% to 12.21%; 86.97% to 91.09%, 2.67% to 8.17%; and 77.86% to 95.36%, 5.02% to 13.15% respectively. The method is simple and sensitive for the quantification of phenolic and salicylanilide anthelmintics in liquid milk.

  13. Analysis of global components in Ganoderma using liquid chromatography system with multiple columns and detectors.

    PubMed

    Qian, Zhengming; Zhao, Jing; Li, Deqiang; Hu, Dejun; Li, Shaoping

    2012-10-01

    In present study, a multiple columns and detectors liquid chromatography system for analysis of global components in traditional Chinese medicines was developed. The liquid chromatography system was consist of three columns, including size exclusion chromatography column, hydrophilic interaction chromatography column, and reversed phase chromatography column, and three detectors, such as diode array detector, evaporative light scattering detector, and mass spectrometry detector, based on column switching technique. The developed multiple columns and detectors liquid chromatography system was successfully applied to the analysis of global components, including macromolecular (polysaccharides), high (nucleosides and sugars)-, and low (triterpenes)-polarity small molecular compounds in Ganoderma, a well-known Chinese medicinal mushroom. As a result, one macromolecular chromatographic peak was found in two Ganoderma species, 19 components were identified in Ganoderma lucidum (two sugars, three nucleosides, and 14 triterpenes), and four components (two sugars and two nucleosides) were identified in Ganoderma sinense. The developed multiple columns and detectors liquid chromatography system was helpful to understand comprehensive chemical characters in TCMs.

  14. Evaluation of a silicon oxynitride hydrophilic interaction liquid chromatography column in saccharide and glycoside separations.

    PubMed

    Wan, Huihui; Sheng, Qianying; Zhong, Hongmin; Guo, Xiujie; Fu, Qing; Liu, Yanfang; Xue, Xingya; Liang, Xinmiao

    2015-05-01

    The retention characteristics of a silicon oxynitride stationary phase for carbohydrate separation were studied in hydrophilic interaction chromatography mode. Four saccharides including mono-, di-, and trisaccharides were employed to investigate the effects of water content and buffer concentration in the mobile phase on hydrophilic interaction liquid chromatography retention. For the tested saccharides, the silicon oxynitride column demonstrated excellent performance in terms of separation efficiency, hydrophilicity, and interesting separation selectivity for carbohydrates compared to the bare silica stationary phase. Finally, the silicon oxynitride hydrophilic interaction liquid chromatography column was employed in the separation of complex samples of fructooligosaccharides, saponins, and steviol glycoside from natural products. The resulting chromatograms demonstrated good separation efficiency and longer retention compared with silica, which further confirmed the advantages and potential application of silicon oxynitride stationary phase for hydrophilic interaction liquid chromatography separation.

  15. Single-Step Process toward Achieving Superhydrophobic Reduced Graphene Oxide.

    PubMed

    Li, Zhong; Tang, Xiu-Zhi; Zhu, Wenyu; Thompson, Brianna C; Huang, Mingyue; Yang, Jinglei; Hu, Xiao; Khor, Khiam Aik

    2016-05-01

    We report the first use of spark plasma sintering (SPS) as a single-step process to achieve superhydrophobic reduced graphene oxide (rGO). It was found that SPS was capable of converting smooth and electrically insulating graphene oxide (GO) sheets into highly electrically conductive rGO with minimum residual oxygen and hierarchical roughness which could be well retained after prolonged ultrasonication. At a temperature of 500 °C, which is lower than the conventional critical temperature for GO exfoliation, GO was successfully exfoliated, reduced, and hierarchically roughened. rGO fabricated by only 1 min of treatment at 1050 °C was superhydrophobic with a surface roughness (Ra) 10 times as large as that of GO as well as an extraordinarily high C:O ratio of 83.03 (atom %) and water contact angle of 153°. This demonstrates that SPS is a superior GO reduction technique, which enabled superhydrophobic rGO to be quickly and effectively achieved in one single step. Moreover, the superhydrophobic rGO fabricated by SPS showed an impressive bacterial antifouling and inactivation effect against Escherichia coli in both aqueous solution and the solid state. It is envisioned that the superhydrophobic rGO obtained in this study can be potentially used for a wide range of industrial and biomedical applications, such as the fabrication of self-cleaning and antibacterial surfaces.

  16. Single-Step Process toward Achieving Superhydrophobic Reduced Graphene Oxide.

    PubMed

    Li, Zhong; Tang, Xiu-Zhi; Zhu, Wenyu; Thompson, Brianna C; Huang, Mingyue; Yang, Jinglei; Hu, Xiao; Khor, Khiam Aik

    2016-05-01

    We report the first use of spark plasma sintering (SPS) as a single-step process to achieve superhydrophobic reduced graphene oxide (rGO). It was found that SPS was capable of converting smooth and electrically insulating graphene oxide (GO) sheets into highly electrically conductive rGO with minimum residual oxygen and hierarchical roughness which could be well retained after prolonged ultrasonication. At a temperature of 500 °C, which is lower than the conventional critical temperature for GO exfoliation, GO was successfully exfoliated, reduced, and hierarchically roughened. rGO fabricated by only 1 min of treatment at 1050 °C was superhydrophobic with a surface roughness (Ra) 10 times as large as that of GO as well as an extraordinarily high C:O ratio of 83.03 (atom %) and water contact angle of 153°. This demonstrates that SPS is a superior GO reduction technique, which enabled superhydrophobic rGO to be quickly and effectively achieved in one single step. Moreover, the superhydrophobic rGO fabricated by SPS showed an impressive bacterial antifouling and inactivation effect against Escherichia coli in both aqueous solution and the solid state. It is envisioned that the superhydrophobic rGO obtained in this study can be potentially used for a wide range of industrial and biomedical applications, such as the fabrication of self-cleaning and antibacterial surfaces. PMID:27064825

  17. Characterization of synthetic dyes by comprehensive two-dimensional liquid chromatography combining ion-exchange chromatography and fast ion-pair reversed-phase chromatography.

    PubMed

    Pirok, Bob W J; Knip, Jitske; van Bommel, Maarten R; Schoenmakers, Peter J

    2016-03-01

    In the late 19th century, newly invented synthetic dyes rapidly replaced the natural dyes on the market. The characterization of mixtures of these so-called early synthetic dyes is complicated through the occurrence of many impurities and degradation products. Conventional one-dimensional liquid chromatography does not suffice to obtain fingerprints with sufficient resolution and baseline integrity. Comprehensive two-dimensional liquid chromatography (LC×LC) is employed in this study, with ion-exchange chromatography in the first dimension and fast ion-pair liquid chromatography in the second. Retention in the first dimension is largely determined by the number of charges, while the selection of a small ion-pair reagent (tetramethylammonium hydroxide) in the second dimension causes retention to be largely determined by the molecular structure of the dye. As a result, there is a high degree of orthogonality of the two dimensions, similar to the values typically encountered in GC×GC. The proposed LC×LC method shows a theroretical peak capacity of about 2000 in an analysis time of about three hours. Clear, informative fingerprints are obtained that open a way to a more efficient characterization of dyes used in objects of cultural heritage.

  18. Characterization of synthetic dyes by comprehensive two-dimensional liquid chromatography combining ion-exchange chromatography and fast ion-pair reversed-phase chromatography.

    PubMed

    Pirok, Bob W J; Knip, Jitske; van Bommel, Maarten R; Schoenmakers, Peter J

    2016-03-01

    In the late 19th century, newly invented synthetic dyes rapidly replaced the natural dyes on the market. The characterization of mixtures of these so-called early synthetic dyes is complicated through the occurrence of many impurities and degradation products. Conventional one-dimensional liquid chromatography does not suffice to obtain fingerprints with sufficient resolution and baseline integrity. Comprehensive two-dimensional liquid chromatography (LC×LC) is employed in this study, with ion-exchange chromatography in the first dimension and fast ion-pair liquid chromatography in the second. Retention in the first dimension is largely determined by the number of charges, while the selection of a small ion-pair reagent (tetramethylammonium hydroxide) in the second dimension causes retention to be largely determined by the molecular structure of the dye. As a result, there is a high degree of orthogonality of the two dimensions, similar to the values typically encountered in GC×GC. The proposed LC×LC method shows a theroretical peak capacity of about 2000 in an analysis time of about three hours. Clear, informative fingerprints are obtained that open a way to a more efficient characterization of dyes used in objects of cultural heritage. PMID:26868445

  19. Supercritical Fluid Chromatography, Pressurized Liquid Extraction and Supercritical Fluid Extraction

    SciTech Connect

    Henry, Matthew C.; Yonker, Clement R.

    2006-06-15

    In this review we examine the related fields of supercritical fluid chromatography (SFC) and supercritical fluid extraction (SFE). We reviewed the published literature in the period from November 2003 to November 2005. Well over 300 papers were published in this period. This large body of work indicates continuing active growth of the field, but an exhaustive review is beyond the scope of this work. We have chosen to include a sampling of publications that best represent the continuing trends and new ideas in the field. In keeping with past reviews on this subject1, we have broadened our scope to include fluid systems operating at high temperature and pressure, but below the critical point. Various terms have been applied to this state: sub-critical fluid extraction, pressurized liquid extraction, and accelerated solvent extraction. The term accelerated solvent extraction has been used by instrument manufacturers to refer to this process, but we will use the more descriptive term pressurized liquid extraction (PLE) to refer to these systems. Most of the research in the field is of an “evolutionary” rather than “revolutionary” nature. As in the previous review period, applications papers make up a majority of the published work. Pharmaceutical applications continue to be a strong theme. Most of the pharmaceutical work has centered on preparative, rather than analytical, separations. Chiral separations are an exception, as analytical scale separations of chiral compounds are an area of intense interest. Food and natural products represent the next largest body of work. Major themes are the isolation and characterization of high-value added foodstuffs, fragrances, and flavor compounds from novel natural materials or agricultural by-products. The areas of food, natural products, and pharmaceutical separation science converge in the area of so-called nutraceuticals. These are typically high-value products, either sold alone or as part of a fortified food, that

  20. High-performance liquid chromatography-ionspray mass spectrometry for the specific determination of digoxin and some related cardiac glycosides in human plasma.

    PubMed

    Tracqui, A; Kintz, P; Ludes, B; Mangin, P

    1997-04-25

    An original method based upon high-performance liquid chromatography coupled to ionspray mass spectrometry (HPLC-ISP-MS) has been developed for the identification and quantification in plasma of several cardiac glycosides, namely digoxin, digitoxin, lanatoside C and acetyldigitoxin. After single-step liquid-liquid extraction by chloroform-2-propanol (95:5, v/v) at pH 9.5 using oleandrin as an internal standard, solutes are separated on a 4 microm NovaPak C18 (Waters) column (150x2.0 mm, I.D.), using a gradient of acetonitrile-2 mM NH4COOH, pH 3 buffer (flow-rate 200 microl/min, post-column split 1:3). Detection is done by a Perkin-Elmer Sciex API-100 mass analyzer equipped with an ISP interface. In most instances the major ion observed is not [M+H]+ as expected, but [M+NH4]+. The mean retention times (min) are: lanatoside C, 5.74; digoxin, 6.00; digitoxin, 8.08, oleandrin, 8.30, acetyldigitoxin, 8.66 and 9.01 (isomers alpha and beta, respectively). The lower limits of detection in single ion monitoring mode range from 0.15 ng/ml (alpha- and beta-acetyldigitoxin) to 0.60 ng/ml (lanatoside C), making the method less sensitive than radioimmunoassay, whereas it is much more specific.

  1. Preparation and evaluation of surface-bonded tricationic ionic liquid silica as stationary phases for high-performance liquid chromatography.

    PubMed

    Qiao, Lizhen; Shi, Xianzhe; Lu, Xin; Xu, Guowang

    2015-05-29

    Two tricationic ionic liquids were prepared and then bonded onto the surface of supporting silica materials through "thiol-ene" click chemistry as new stationary phases for high-performance liquid chromatography. The obtained columns of tricationic ionic liquids were evaluated respectively in the reversed-phase liquid chromatography (RPLC) mode and hydrophilic interaction liquid chromatography (HILIC) mode, and possess ideal column efficiency of 80,000 plates/m in the RPLC mode with naphthalene as the test solute. The tricationic ionic liquid stationary phases exhibit good hydrophobic and shape selectivity to hydrophobic compounds, and RPLC retention behavior with multiple interactions. In the HILIC mode, the retention and selectivity were evaluated through the efficient separation of nucleosides and bases as well as flavonoids, and the typical HILIC retention behavior was demonstrated by investigating retention changes of hydrophilic solutes with water volume fraction in mobile phase. The results show that the tricationic ionic liquid columns possess great prospect for applications in analysis of hydrophobic and hydrophilic samples. PMID:25890438

  2. Preparation and evaluation of surface-bonded tricationic ionic liquid silica as stationary phases for high-performance liquid chromatography.

    PubMed

    Qiao, Lizhen; Shi, Xianzhe; Lu, Xin; Xu, Guowang

    2015-05-29

    Two tricationic ionic liquids were prepared and then bonded onto the surface of supporting silica materials through "thiol-ene" click chemistry as new stationary phases for high-performance liquid chromatography. The obtained columns of tricationic ionic liquids were evaluated respectively in the reversed-phase liquid chromatography (RPLC) mode and hydrophilic interaction liquid chromatography (HILIC) mode, and possess ideal column efficiency of 80,000 plates/m in the RPLC mode with naphthalene as the test solute. The tricationic ionic liquid stationary phases exhibit good hydrophobic and shape selectivity to hydrophobic compounds, and RPLC retention behavior with multiple interactions. In the HILIC mode, the retention and selectivity were evaluated through the efficient separation of nucleosides and bases as well as flavonoids, and the typical HILIC retention behavior was demonstrated by investigating retention changes of hydrophilic solutes with water volume fraction in mobile phase. The results show that the tricationic ionic liquid columns possess great prospect for applications in analysis of hydrophobic and hydrophilic samples.

  3. Crosslinked polymeric ionic liquids as solid-phase microextraction sorbent coatings for high performance liquid chromatography.

    PubMed

    Yu, Honglian; Merib, Josias; Anderson, Jared L

    2016-03-18

    Neat crosslinked polymeric ionic liquid (PIL) sorbent coatings for solid-phase microextraction (SPME) compatible with high-performance liquid chromatography (HPLC) are reported for the first time. Six structurally different PILs were crosslinked to nitinol supports and applied for the determination of select pharmaceutical drugs, phenolics, and insecticides. Sampling conditions including sample solution pH, extraction time, desorption solvent, desorption time, and desorption solvent volume were optimized using design of experiment (DOE). The developed PIL sorbent coatings were stable when performing extractions under acidic pH and remained intact in various organic desorption solvents (i.e., methanol, acetonitrile, acetone). The PIL-based sorbent coating polymerized from the IL monomer 1-vinyl-3-(10-hydroxydecyl) imidazolium chloride [VC10OHIM][Cl] and IL crosslinker 1,12-di(3-vinylbenzylimidazolium) dodecane dichloride [(VBIM)2C12] 2[Cl] exhibited superior extraction performance compared to the other studied PILs. The extraction efficiency of pharmaceutical drugs and phenolics increased when the film thickness of the PIL-based sorbent coating was increased while many insecticides were largely unaffected. Satisfactory analytical performance was obtained with limits of detection (LODs) ranging from 0.2 to 2 μg L(-1) for the target analytes. The accuracy of the analytical method was examined by studying the relative recovery of analytes in real water samples, including tap water and lake water, with recoveries varying from 50.2% to 115.9% and from 48.8% to 116.6%, respectively. PMID:26896916

  4. Crosslinked polymeric ionic liquids as solid-phase microextraction sorbent coatings for high performance liquid chromatography.

    PubMed

    Yu, Honglian; Merib, Josias; Anderson, Jared L

    2016-03-18

    Neat crosslinked polymeric ionic liquid (PIL) sorbent coatings for solid-phase microextraction (SPME) compatible with high-performance liquid chromatography (HPLC) are reported for the first time. Six structurally different PILs were crosslinked to nitinol supports and applied for the determination of select pharmaceutical drugs, phenolics, and insecticides. Sampling conditions including sample solution pH, extraction time, desorption solvent, desorption time, and desorption solvent volume were optimized using design of experiment (DOE). The developed PIL sorbent coatings were stable when performing extractions under acidic pH and remained intact in various organic desorption solvents (i.e., methanol, acetonitrile, acetone). The PIL-based sorbent coating polymerized from the IL monomer 1-vinyl-3-(10-hydroxydecyl) imidazolium chloride [VC10OHIM][Cl] and IL crosslinker 1,12-di(3-vinylbenzylimidazolium) dodecane dichloride [(VBIM)2C12] 2[Cl] exhibited superior extraction performance compared to the other studied PILs. The extraction efficiency of pharmaceutical drugs and phenolics increased when the film thickness of the PIL-based sorbent coating was increased while many insecticides were largely unaffected. Satisfactory analytical performance was obtained with limits of detection (LODs) ranging from 0.2 to 2 μg L(-1) for the target analytes. The accuracy of the analytical method was examined by studying the relative recovery of analytes in real water samples, including tap water and lake water, with recoveries varying from 50.2% to 115.9% and from 48.8% to 116.6%, respectively.

  5. Single Step Endodontic Management of Immature Apices using MTA

    PubMed Central

    Mishra, SK; Kumar, Ashok; Garg, Amit Kumar; Jindal, MK; Juneja, Harish

    2011-01-01

    Aim To examine the clinical and radiographic appearance of teeth with immature apices that were treated by single step procedure using mineral trioxide aggregate (MTA). Summary Creation of a physiological hard tissue barrier with calcium hydroxide in a nonvital tooth although quite predictable has certain limitations, such as the very long duration of the treatment spread over multiple visits and increased risk of root fracture. Plugging the root canal end with MTA has been advocated as an alternative treatment modality for open apices. The technique has been proven to be successful in many recently reported cases. The cases reported here present the successful treatment of two traumatized maxillary central incisors with open apices and periapical lesions using MTA. In this case report, MTA has been used to create a hard tissue barrier after disinfection of the root canal.

  6. Algal-based, single-step treatment of urban wastewaters.

    PubMed

    Henkanatte-Gedera, S M; Selvaratnam, T; Caskan, N; Nirmalakhandan, N; Van Voorhies, W; Lammers, Peter J

    2015-08-01

    Currently, urban wastewaters (UWW) laden with organic carbon (BOD) and nutrients (ammoniacal nitrogen, N, and phosphates, P) are treated in multi-stage, energy-intensive process trains to meet the mandated discharge standards. This study presents a single-step process based on mixotrophic metabolism for simultaneous removal of carbon and nutrients from UWWs. The proposed system is designed specifically for hot, arid environments utilizing an acidophilic, thermotolerant algal species, Galdieria sulphuraria, and an enclosed photobioreactor to limit evaporation. Removal rates of BOD, N, and P recorded in this study (14.93, 7.23, and 1.38 mg L(-1) d(-1), respectively) are comparable to literature reports. These results confirm that the mixotrophic system can reduce the energy costs associated with oxygen supply in current UWW treatment systems, and has the potential to generate more energy-rich biomass for net energy extraction from UWW.

  7. Single Step Endodontic Management of Immature Apices using MTA

    PubMed Central

    Mishra, SK; Kumar, Ashok; Garg, Amit Kumar; Jindal, MK; Juneja, Harish

    2011-01-01

    Aim To examine the clinical and radiographic appearance of teeth with immature apices that were treated by single step procedure using mineral trioxide aggregate (MTA). Summary Creation of a physiological hard tissue barrier with calcium hydroxide in a nonvital tooth although quite predictable has certain limitations, such as the very long duration of the treatment spread over multiple visits and increased risk of root fracture. Plugging the root canal end with MTA has been advocated as an alternative treatment modality for open apices. The technique has been proven to be successful in many recently reported cases. The cases reported here present the successful treatment of two traumatized maxillary central incisors with open apices and periapical lesions using MTA. In this case report, MTA has been used to create a hard tissue barrier after disinfection of the root canal. PMID:27678239

  8. Isolation and purification of six iridoid glycosides from gardenia jasminoides fruit by medium-pressure liquid chromatography combined with macroporous resin chromatography.

    PubMed

    Wang, Yun; Liu, Hui; Shen, Lifeng; Yao, Lan; Ma, Yinlian; Yu, Dingrong; Chen, Jianhong; Li, Puling; Chen, Ying; Zhang, Cun

    2015-12-01

    Gardeniae fructus is one of the most frequently used herbs in traditional Chinese medicine. In the present study, a process for the enrichment of six iridoid glycosides from Gardeniae fructus was developed using medium-pressure liquid chromatography combined with macroporous resin and reversed-phase chromatography. The purities of different fractions from Gardeniae fructus were assessed using quantitative high-performance liquid chromatography. After fractionation using HPD-100 column chromatography, a 30% ethanol fraction was selected based on high-performance liquid chromatography and liquid chromatography with mass spectrometry qualitative analysis to separate and purify. Based on the orientation analysis results, six compounds-deacetyl asperulosidic acid methyl ester, gardenoside, ixoroside, scandoside methyl ester, genipin-1-O-β-d-gentiobioside, and geniposide-were successfully isolated and purified in three to four combined steps from Gardeniae fructus. The purities of these compounds were found by high-performance liquid chromatography analysis to be 97.9, 98.1, 95.5, 96.3, 97.1, and 98.7%, respectively. Moreover, their structures were elucidated by NMR spectroscopy and liquid chromatography with tandem mass spectrometry. The separation process was highly efficient, rapid, and accurate, making it a potential approach for the large-scale production of iridoids in the laboratory and providing several marker compounds for quality control. This procedure may be meaningful for the purification of other natural products used in traditional Chinese medicine. PMID:26768549

  9. Isolation and purification of six iridoid glycosides from gardenia jasminoides fruit by medium-pressure liquid chromatography combined with macroporous resin chromatography.

    PubMed

    Wang, Yun; Liu, Hui; Shen, Lifeng; Yao, Lan; Ma, Yinlian; Yu, Dingrong; Chen, Jianhong; Li, Puling; Chen, Ying; Zhang, Cun

    2015-12-01

    Gardeniae fructus is one of the most frequently used herbs in traditional Chinese medicine. In the present study, a process for the enrichment of six iridoid glycosides from Gardeniae fructus was developed using medium-pressure liquid chromatography combined with macroporous resin and reversed-phase chromatography. The purities of different fractions from Gardeniae fructus were assessed using quantitative high-performance liquid chromatography. After fractionation using HPD-100 column chromatography, a 30% ethanol fraction was selected based on high-performance liquid chromatography and liquid chromatography with mass spectrometry qualitative analysis to separate and purify. Based on the orientation analysis results, six compounds-deacetyl asperulosidic acid methyl ester, gardenoside, ixoroside, scandoside methyl ester, genipin-1-O-β-d-gentiobioside, and geniposide-were successfully isolated and purified in three to four combined steps from Gardeniae fructus. The purities of these compounds were found by high-performance liquid chromatography analysis to be 97.9, 98.1, 95.5, 96.3, 97.1, and 98.7%, respectively. Moreover, their structures were elucidated by NMR spectroscopy and liquid chromatography with tandem mass spectrometry. The separation process was highly efficient, rapid, and accurate, making it a potential approach for the large-scale production of iridoids in the laboratory and providing several marker compounds for quality control. This procedure may be meaningful for the purification of other natural products used in traditional Chinese medicine.

  10. Speciation analysis of mercury in water samples using dispersive liquid-liquid microextraction combined with high-performance liquid chromatography.

    PubMed

    Gao, Zhongben; Ma, Xiaoguo

    2011-09-19

    A novel approach for preconcentration and speciation analysis of trace amount of mercury from water samples was proposed by dispersive liquid-liquid microextraction (DLLME) coupled to high performance liquid chromatography with diode array detection (HPLC-DAD). Mercury species (Hg(2+), methylmercury (MeHg(+)) and phenylmercury (PhHg(+))) were complexed with dithizone (DZ) to form hydrophobic chelates and then extracted into the fine drops of extraction solvent dispersed in the aqueous sample by dispersive solvent. After extraction, the sedimented phase was analyzed by HPLC-DAD. Some important parameters affecting the DLLME such as extraction solvent and dispersive solvent type and volume, concentration of dithizone solution, sample pH, extraction time and salt effect were investigated. Ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIM][PF(6)]) was found to be a suitable extractant for the chelates. Under the optimized conditions (extraction solvent: 70 μL of ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate ([HMIM][PF(6)]); dispersive solvent: 0.75 mL of methanol containing dithizone (0.02%, m/v); pH: 4; extraction time: 5 min; and without salt addition), the limits of detection for Hg(2+), MeHg(+) and PhHg(+) were 0.32, 0.96 and 1.91 μg L(-1) (SN(-1)=3) respectively, and the relative standard deviation (RSD) was between 4.1 and 7.3% (n=5). Three real water samples (tap water, river water and lake water) spiked with mercury species were detected by the developed method, and the relative recoveries obtained for Hg(2+), MeHg(+) and PhHg(+) were 89.6-101.3%, 85.6-102.0% and 81.3-97.6%, respectively. PMID:21819859

  11. Monodisperse microbeads of hypercrosslinked polystyrene for liquid and supercritical fluid chromatography

    NASA Astrophysics Data System (ADS)

    Tsyurupa, M. P.; Blinnikova, Z. K.; Il'in, M. M.; Davankov, V. A.; Parenago, O. O.; Pokrovskii, O. I.; Usovich, O. I.

    2015-11-01

    Monodisperse styrene-divinylbenzene (1 wt %) copolymer microbeads are obtained via the elaborate method of high-productivity precipitation polymerization. The crosslinking of this copolymer with chloromethyl methyl ether in the presence of Friedel-Crafts catalyst yields porous hypercrosslinked polymers with degrees of crosslinking that range from 200 to 500%. Microbead sorbents are shown to be suited for selective stationary phases for high-performance liquid chromatography and supercritical fluid chromatography.

  12. Surface diffusion in reversed-phase liquid chromatography

    SciTech Connect

    Miyabe, Kanji; Guiochon, Georges A

    2010-01-01

    More than 40 years ago, Giddings pointed out in 'Dynamics of Chromatography' that surface diffusion should become an important research topic in the kinetics of chromatographic phenomena. However, few studies on surface diffusion in adsorbents used in chromatography were published since then. Most scientists use ordinary rate equations to study mass transfer kinetics in chromatography. They take no account of surface diffusion and overlook the significant contributions of this mass transfer process to chromatographic behavior and to column efficiency at high mobile phase flow rate. Only recently did the significance of surface diffusion in separation processes begin to be recognized in connection with the development of new techniques of fast flow, high efficiency chromatography. In this review, we revisit the reports on experimental data on surface diffusion and introduce a surface-restricted molecular diffusion model, derived as a first approximation for the mechanism of surface diffusion, on the basis of the absolute rate theory. We also explain how this model accounts for many intrinsic characteristics of surface diffusion that cannot properly be explained by the conventional models of surface diffusion.

  13. Vortex-assisted liquid-liquid-liquid microextraction (VALLLME) technique: A new microextraction approach for direct liquid chromatography and capillary electrophoresis analysis.

    PubMed

    Makahleh, Ahmad; Yap, Hui Fang; Saad, Bahruddin

    2015-10-01

    A new, rapid and sensitive microextraction technique named vortex-assisted liquid-liquid-liquid microextraction (VALLLME) is proposed. The complete extraction process involves two steps. First, a vortex-assisted liquid-liquid microextraction (VALLME) procedure was used to extract the analytes from a relatively large volume of sample (donor phase) to a small volume of organic solvent (intermediate phase). Next, a micro-vortex-assisted liquid-liquid extraction (µ-VALLE) was used to extract the target analytes from the intermediate phase to a smaller volume of aqueous solution (acceptor phase). The final extract (acceptor phase) can be directly injected into the high performance liquid chromatography or capillary electrophoresis units without any further treatments. The selection of the intermediate phase and the manipulation of pH are key parameters that ensure good extraction efficiency of the technique. The proposed technique has been successfully applied for the determination of carvedilol (used as model analyte) in biological fluid samples. The optimum extraction conditions were: toluene as intermediate phase (150 μL); pH of the donor phase, 9.5; vortex time of the VALLME, 45 s (maximum speed, 2500 rpm); 0.1M HCl (15 μL) as acceptor phase; vortexing time of the µ-VALLME, 75 s (maximum stirring speed, 2500 rpm) and salt concentration in the donor phase, 5% (w/v). Under these conditions, enrichment factors of 51- and 418-fold for VALLME step and VALLLME procedure, respectively, were achieved. PMID:26078176

  14. Comprehensive Microstructure and Molar Mass Analysis of Polybutadiene by Multidimensional Liquid Chromatography.

    PubMed

    Maiko, Khumo; Pasch, Harald

    2015-12-01

    For the first time, polybutadiene is separated according to microstructure using solvent gradient interaction chromatography (SGIC). Superior separation of polybutadienes having different microstructures is obtained on a silica-based reversed stationary phase and a mobile phase of acetone-hexane. This SGIC system enables the baseline separation of 1,2-polybutadienes and 1,4-polybutadienes even in cases where the samples have similar molar masses. 2D liquid chromatography is performed with the SGIC method separating according to microstructure in the first dimension coupled to size exclusion chromatography separating according to molar mass in the second dimension, thus providing comprehensive information on both microstructure and molar mass.

  15. Comprehensive Microstructure and Molar Mass Analysis of Polybutadiene by Multidimensional Liquid Chromatography.

    PubMed

    Maiko, Khumo; Pasch, Harald

    2015-12-01

    For the first time, polybutadiene is separated according to microstructure using solvent gradient interaction chromatography (SGIC). Superior separation of polybutadienes having different microstructures is obtained on a silica-based reversed stationary phase and a mobile phase of acetone-hexane. This SGIC system enables the baseline separation of 1,2-polybutadienes and 1,4-polybutadienes even in cases where the samples have similar molar masses. 2D liquid chromatography is performed with the SGIC method separating according to microstructure in the first dimension coupled to size exclusion chromatography separating according to molar mass in the second dimension, thus providing comprehensive information on both microstructure and molar mass. PMID:26461063

  16. On-line comprehensive two-dimensional normal-phase liquid chromatography×reversed-phase liquid chromatography for preparative isolation of toad venom.

    PubMed

    Li, Jia-Fu; Fang, Hua; Yan, Xia; Chang, Fang-Rong; Wu, Zhen; Wu, Yun-Long; Qiu, Ying-Kun

    2016-07-22

    An on-line comprehensive preparative two-dimensional normal-phase liquid chromatography×reversed-phase liquid chromatography (2D NPLC×RPLC) system was constructed with a newly developed vacuum evaporation assisted adsorption (VEAA) interface, allowing fast removal of NPLC solvent in the vacuum condition and successfully solving the solvent incompatibility problem between NPLC and RPLC. The system achieved on-line solvent exchange within the two dimensions and its performance was illustrated by gram-scale isolation of crude extract from the venom of Bufo bufo gargarizans. Within separation time of ∼20h, 19 compounds were obtained with high purity in a single run. With the VEAA interface, the 2D system exhibited apparent advantages in separation efficiency and automation compared with conventional methods, indicating its promising application in the routine separation process for complicated natural products. PMID:27328884

  17. High-pressure liquid chromatography: A brief introduction and its application in analyzing the degradation of a C-ether (Thio-ether) liquid lubricant

    NASA Technical Reports Server (NTRS)

    1983-01-01

    The general principles of classical liquid chromatography and high pressure liquid chromatography (HPLC) are reviewed, and their advantages and disadvantages are compared. Several chromatographic techniques are reviewed, and the analytical separation of a C-ether liquid lubricant by each technique is illustrated. A practical application of HPLC is then demonstrated by analyzing a degraded C-ether liquid lubricant from full scale, high temperature bearing tests.

  18. High-performance liquid chromatography of oligoguanylates at high pH

    NASA Technical Reports Server (NTRS)

    Stribling, R.; Deamer, D. (Principal Investigator)

    1991-01-01

    Because of the stable self-structures formed by oligomers of guanosine, standard high-performance liquid chromatography techniques for oligonucleotide fractionation are not applicable. Previously, oligoguanylate separations have been carried out at pH 12 using RPC-5 as the packing material. While RPC-5 provides excellent separations, there are several limitations, including the lack of a commercially available source. This report describes a new anion-exchange high-performance liquid chromatography method using HEMA-IEC BIO Q, which successfully separates different forms of the guanosine monomer as well as longer oligoguanylates. The reproducibility and stability at high pH suggests a versatile role for this material.

  19. Automated dispersive liquid-liquid microextraction-gas chromatography-mass spectrometry.

    PubMed

    Guo, Liang; Lee, Hian Kee

    2014-04-15

    An innovative automated procedure, low-density solvent based/solvent demulsification dispersive liquid-liquid microextraction (automated DLLME) coupled to gas chromatography-mass spectrometry (GC/MS) analysis, has been developed. The most significant innovation of the method is the automation. The entire procedure, including the extraction of the model analytes (phthalate esters) by DLLME from the aqueous sample solution, breaking up of the emulsion after extraction, collection of the extract, and analysis of the extract by GC/MS, was completely automated. The applications of low-density solvent as extraction solvent and the solvent demulsification technique to break up the emulsion simplified the procedure and facilitated its automation. Orthogonal array design (OAD) as an efficient optimization strategy was employed to optimize the extraction parameters, with all the experiments conducted auotmatically. An OA16 (4(1) × 2(12)) matrix was initially employed for the identification of optimized extraction parameters (type and volume of extraction solvent, type and volume of dispersive solvent and demulsification solvent, demulsification time, and injection speed). Then, on the basis of the results, more levels (values) of five extraction parameters were investigated by an OA16 (4(5)) matrix and quantitatively assessed by the analysis of variance (ANOVA). Enrichment factors of between 178- and 272-fold were obtained for the phthalate esters. The linearities were in the range of 0.1 and 50 μg/L and 0.2 and 50 μg/L, depending on the analytes. Good limits of detection (in the range of 0.01 to 0.02 μg/L) and satisfactory repeatability (relative standard deviations of below 5.9%) were obtained. The proposed method demonstrates for the first time integrated sample preparation by DLLME and analysis by GC/MS that can be operated automatically across multiple experiments.

  20. Determination of organophosphorus pesticides in soil by dispersive liquid-liquid microextraction and gas chromatography.

    PubMed

    Yang, Zhonghua; Liu, Yu; Liu, Donghui; Zhou, Zhiqiang

    2012-01-01

    In this article, a rapid and sensitive sample pretreatment technique for the determination of organophosphorus pesticides (OPPs) in soil samples is developed by using dispersive liquid-liquid microextraction (DLLME) combined with gas chromatography-flame photometric detection. Experimental conditions, including the kind of extraction and disperser solvent and their volumes, the extraction time, and the salt addition, are investigated, and the following experiment factors are used: 20 µL chlorobenzene as the extraction solvent; 1.0 mL acetonitrile as the disperser solvent; no addition of salt; and an extraction time of 1 min. Under the optimum conditions, the linearities for the three target OPPs (ethoprophos, chlorpyriphos, and profenofos) are obtained by five points in the concentration range of 2.5-1500 µg/kg, and three replicates are used for each point. Correlation coefficients vary from 0.9987 to 0.9997. The repeatability is tested by spiking soil samples at a concentration level of 5.0 µg/kg. The relative standard deviation (n = 3) varied between 2.0% and 6.6%. The limits of detection, based on a signal-to-noise ratio (S/N) of 3, range from 200 to 500 pg/g. This method is applied to the analysis of the spiked samples S1, S2, and S3, which are collected from the China Agriculture University's orchard, lawn, and garden, respectively. The recoveries for each target analyte are in the range between 87.9% and 108.0%, 87.4% and 108.0%, and 86.7% and 107.2%, respectively. PMID:22291051

  1. Single-step PCR in molecular diagnosis of hepatitis C virus infection.

    PubMed

    Farma, E; Boeri, E; Bettini, P; Repetto, C M; McDermott, J; Lillo, F B; Varnier, O E

    1996-12-01

    The diagnostic utility of two PCR systems and three PCR detection methods for hepatitis C virus (HCV) RNA was evaluated in serum samples. A nested PCR was considered the reference assay and was compared with two single-step PCR methods: the first is based on the detection of PCR products by liquid hybridization with a 32P-end-labeled probe, and the second is the Roche Amplicor colorimetric assay using microwell plate hybridization with a specific nucleic acid probe. Using the Pelicheck HCV RNA Eurohep genotype 1 proficiency panel, our laboratory achieved medium-high levels of performance with all three methods. The highest sensitivity was, however, observed with the isotopic single-step PCR (ss-PCR) method. The analytical sensitivity of ss-PCR with isotopic detection and ss-PCR with colorimetric detection was identical to that of nested PCR, with a 100% result concordance. Comparison of ss-PCR with enzyme-linked immunosorbent and RIBA assays in the analysis of clinical samples showed a high concordance. ss-PCR methods appear more suitable for diagnostic application. Nevertheless, HCV RNA PCR cannot be considered a screening assay; it should be requested in the presence of reactive serology or specific clinical symptomatology with altered liver parameters, and it is a potential tool for the follow-up of patients with HCV infection.

  2. Identification of Explosives from Porous Materials: Applications Using Reverse Phase High Performance Liquid Chromatography and Gas Chromatography

    SciTech Connect

    C.J. Miller; G. Elias; N.C. Schmitt; C. Rae

    2010-06-01

    High performance liquid chromatography and gas chromatography techniques are well documented and widely used for the detection of trace explosives from organic solvents. These techniques were modified to specifically identify and quantify explosives extracted from various materials taken from people who had recently handled explosives. Documented techniques were modified to specifically detect and quantify RDX, TNT, and PETN from denim, colored flannel, vinyl, and canvas extracted in methanol using no sample cleanup prior to analysis. The methanol extracts were injected directly into several different column types and analyzed by HPLC-UV and/or GC-ECD. This paper describes general screening methods that were used to determine the presence of explosives in unknown samples and techniques that have been optimized for quantification of each explosive from the substrate extracts.

  3. Thin-layer chromatography-densitometry and liquid chromatography analysis of alkaloids in leaves of Papaver somniferum under stress conditions.

    PubMed

    Szabó, Beata; Lakatos, Agnes; Kõszegi, Tamás; Kátay, György; Botz, L

    2005-01-01

    The effect of stress conditions on the concentrations of secondary metabolites were examined during various developmental stages of Papaver somniferum plants. P. somniferum plants were grown in laboratory conditions (Budakalász). The experiment consisted of 22 treatments. Significantly different alkaloid contents can be observed under different stress conditions. In general, the alkaloid contents of plants are very low; therefore, a highly sensitive and reliable method has to be developed for analysis. The amount of alkaloids was measured by 2 separation and detection techniques. Accuracy of the thin-layer chromatography method for quantitative analysis is limited. Without purification of samples the background is too noisy. Column liquid chromatography is a sensitive and relatively inexpensive method that allows precise quantitative determination of the alkaloid content.

  4. Strong cation exchange chiral stationary phase--a comparative study in high-performance liquid chromatography and subcritical fluid chromatography.

    PubMed

    Wolrab, Denise; Macíková, Pavla; Boras, Mario; Kohout, Michal; Lindner, Wolfgang

    2013-11-22

    The performance of a strong cation exchange-type (SCX) chiral stationary phase (CSP) was evaluated with subcritical fluid chromatography (subFC) and high performance liquid chromatography (HPLC). The chromatographic conditions in subFC were optimized by changing the amount of polar organic modifier, concentration of a basic additive in the modifier, system pressure and temperature. In this way the concentration of in situ formed transient ionic species could be varied. The gradual change of the concentration of the transient buffer, i.e. gradient elution conditions in subFC, was found beneficial for separation of a mixture of racemic compounds. The strength and amount of the in situ formed buffer was estimated on the basis of comparative experiments in subSFC and HPLC.

  5. Ultrasound-assisted ionic liquid-based homogeneous liquid-liquid microextraction high-performance liquid chromatography for determination of tanshinones in Salvia miltiorrhiza Bge. root.

    PubMed

    Wang, Zhibing; Cao, Bocheng; Yu, Aimin; Zhang, Hanqi; Qiu, Fangping

    2015-02-01

    The ultrasound-assisted ionic liquid-based homogeneous liquid-liquid microextraction has been developed and applied to the extraction of four tanshinones, including dihydrotanshinone, tanshinone I, cryptotanshinone and tanshinone IIA in Salvia miltiorrhiza Bge. root. High performance liquid chromatography was applied to the separation and determination of the analytes. The ionic liquid was used as extraction solvent and target analytes were extracted with help of ultrasound. Then, ion-pairing agent was added into the sample solution, which resulted in the formation of water-insoluble ionic liquid in the solution. The phase separation was performed by centrifugation. The extraction, concentration and purification of target analytes were performed simultaneously. The experimental parameters, including type and volume of ionic liquid, sample amount, the size of sample particle, pH value of extraction medium, extraction temperature, extraction time, amount of ion-pairing agent and centrifuging time, were investigated and optimized. The calibration curves showed good linear relationship (r>0.9997). The limits of detection and quantification were in the range of 0.052-0.093 and 0.17-0.31 μg mL(-1), respectively. The recoveries were between 70.45% and 94.23% with relative standard deviations lower than 5.31%. The present method is free of volatile organic solvents, and represents lower expenditures of sample, extraction time and solvent, compared with UAE and HRE. There was no obvious difference in the extraction yields of active constitutions obtained by the three extraction methods.

  6. Single step radiolytic synthesis of iridium nanoparticles onto graphene oxide

    NASA Astrophysics Data System (ADS)

    Rojas, J. V.; Molina Higgins, M. C.; Toro Gonzalez, M.; Castano, C. E.

    2015-12-01

    In this work a new approach to synthesize iridium nanoparticles on reduced graphene oxide is presented. The nanoparticles were directly deposited and grown on the surface of the carbon-based support using a single step reduction method through gamma irradiation. In this process, an aqueous isopropanol solution containing the iridium precursor, graphene oxide, and sodium dodecyl sulfate was initially prepared and sonicated thoroughly to obtain a homogeneous dispersion. The samples were irradiated with gamma rays with energies of 1.17 and 1.33 MeV emitted from the spontaneous decay of the 60Co irradiator. The interaction of gamma rays with water in the presence of isopropanol generates highly reducing species homogeneously distributed in the solution that can reduce the Ir precursor down to a zero valence state. An absorbed dose of 60 kGy was used, which according to the yield of reducing species is sufficient to reduce the total amount of precursor present in the solution. This novel approach leads to the formation of 2.3 ± 0.5 nm Ir nanoparticles distributed along the surface of the support. The oxygenated functionalities of graphene oxide served as nucleation sites for the formation of Ir nuclei and their subsequent growth. XPS results revealed that the interaction of Ir with the support occurs through Irsbnd O bonds.

  7. Determination of tetraalkyllead compounds in gasoline by liquid chromatography-atomic absorption spectrometry

    USGS Publications Warehouse

    Messman, J.D.; Rains, T.C.

    1981-01-01

    A liquid chromatography-atomic absorption spectrometry (LC-AAS) hybrid analytical technique is presented for metal speciation measurements on complex liquid samples. The versatility and inherent metal selectivity of the technique are Illustrated by the rapid determination of five tetraalkyllead compounds in commercial gasoline. Separation of the individual tetraalkyllead species is achieved by reversed-phase liquid chromatography using an acetonitrile/water mobile phase. The effluent from the liquid Chromatograph Is introduced directly into the aspiration uptake capillary of the nebulizer of an air/acetylene flame atomic absorption spectrometer. Spectral interferences due to coeluting hydrocarbon matrix constituents were not observed at the 283.3-nm resonance line of lead used for analysis. Detection limits of this LC-AAS hydrid analytical technique, based on a 20-??L injection, are approximately 10 ng Pb for each tetraalkyllead compound.

  8. DETERMINATION OF CARBENDAZIM IN WATER BY HIGH-PERFORMANCE IMMUNOAFFINITY CHROMATOGRAPHY ON-LINE WITH HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH DIODE-ARRAY OR MASS SPECTROMETRIC DETECTION

    EPA Science Inventory

    An automated method for the determination of carbendazim in water that combines high-performance immunoaffinity chromatography (HPIAC), high-performance liquid chromatography (HPLC) in the reversed-phase mode, and detection by either UV-Vis diode array detector (DAD) spectroscopy...

  9. Application in pesticide analysis: Liquid chromatography - A review of the state of science for biomarker discovery and identification

    EPA Science Inventory

    Book Chapter 18, titled Application in pesticide analysis: Liquid chromatography - A review of the state of science for biomarker discovery and identification, will be published in the book titled High Performance Liquid Chromatography in Pesticide Residue Analysis (Part of the C...

  10. Determination of five antiarrhythmic drugs in human plasma by dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    PubMed

    Jouyban, Abolghasem; Sorouraddin, Mohammad Hossein; Farajzadeh, Mir Ali; Somi, Mohammad Hossein; Fazeli-Bakhtiyari, Rana

    2015-03-01

    A fast and sensitive high-performance liquid chromatography (HPLC) method with ultraviolet (UV) detection was developed and validated for the simultaneous quantitation of five antiarrhythmic drugs (metoprolol, propranolol, carvedilol, diltiazem, and verapamil) in human plasma samples. It involves dispersive liquid-liquid microextraction (DLLME) of the desired drugs from 660 µL plasma and separation using isocratic elution with UV detection at 200 nm. The complete separation of all analytes was achieved within 7 min. Acetonitrile (as disperser solvent) resulting from the protein precipitation procedure was mixed with 100 µL dichloromethane (as an extraction solvent) and rapidly injected into 5 mL aqueous solution (pH 11.5) containing 1% (w/v), NaCl. After centrifugation, the sedimented phase containing enriched analytes was collected and evaporated to dryness. The residue was re-dissolved in 50 µL de-ionized water (acidified to pH 3) and injected into the HPLC system for analysis. Under the optimal conditions, the enrichment factors and extraction recoveries ranged between 4.4-10.8 and 33-82%, respectively. The suggested method was linear (r(2) ≥0.997) over a dynamic range of 0.02-0.80 µg mL(-1) in plasma. The intra- and inter-days relative standard deviation (RSD%) and relative error (RE%) values of the method were below 20%, which shows good precision and accuracy. Finally, this method was applied to the analysis of real plasma samples obtained from the patients treated with these drugs.

  11. Analysis of drugs of abuse in human plasma by dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    PubMed

    Fernández, P; Regenjo, M; Bermejo, A M; Fernández, A M; Lorenzo, R A; Carro, A M

    2015-04-01

    Opioids and cocaine are widely used at present, both for recreational purposes and as drugs of abuse. This raises the need to develop new analytical methods specifically designed for the simultaneous detection of several drugs of abuse in biological samples. In this work, dispersive liquid-liquid microextraction (DLLME) was assessed as a new sample treatment for the simultaneous extraction of morphine (MOR), 6-acetylmorphine (6AM), cocaine (COC), benzoylecgonine (BZE) and methadone (MET) from human plasma. Preliminary assays were done before developing an experimental design based on a Uniform Network Doehlert which allowed the optimum extraction conditions to be identified, namely: a volume of extractant solvent (chloroform) and dispersant solvent (acetonitrile) of 220 µl and 3.2 ml, respectively; 0.2 g of NaCl as a salting-out additive; pH 10.6 and ultrasound stirring for 3.5 min. The resulting extracts were analyzed by high-performance liquid chromatography with photodiode array detection (HPLC-PDA), using an XBridge® RP18 column (250 × 4.6 mm i.d., 5 µm particle size). Calibration graphs were linear over the concentration range 0.1-10 µg ml⁻¹, and detection limits ranged from 13.9 to 28.5 ng ml⁻¹. Precision calculated at three different concentration levels in plasma was included in the range 0.1-6.8% RSD. Recoveries of the five drugs were all higher than 84% on average. Finally the proposed method was successfully applied to 22 plasma samples from heroin, cocaine and/or methadone users, and the most frequently detected drug was benzoylecgonine, followed by methadone, cocaine and morphine.

  12. Liquid chromatography and supercritical fluid chromatography as alternative techniques to gas chromatography for the rapid screening of anabolic agents in urine.

    PubMed

    Desfontaine, Vincent; Nováková, Lucie; Ponzetto, Federico; Nicoli, Raul; Saugy, Martial; Veuthey, Jean-Luc; Guillarme, Davy

    2016-06-17

    This work describes the development of two methods involving supported liquid extraction (SLE) sample treatment followed by ultra-high performance liquid chromatography or ultra-high performance supercritical fluid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS and UHPSFC-MS/MS) for the screening of 43 anabolic agents in human urine. After evaluating different stationary phases, a polar-embedded C18 and a diol columns were selected for UHPLC-MS/MS and UHPSFC-MS/MS, respectively. Sample preparation, mobile phases and MS conditions were also finely tuned to achieve highest selectivity, chromatographic resolution and sensitivity. Then, the performance of these two methods was compared to the reference routine procedure for steroid analyses in anti-doping laboratories, which combines liquid-liquid extraction (LLE) followed by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). For this purpose, urine samples spiked with the compounds of interest at five different concentrations were analyzed using the three analytical platforms. The retention and selectivity of the three techniques were very different, ensuring a good complementarity. However, the two new methods displayed numerous advantages. The overall procedure was much faster thanks to high throughput SLE sample treatment using 48-well plates and faster chromatographic analysis. Moreover, the highest sensitivity was attained using UHPLC-MS/MS with 98% of the doping agents detected at the lowest concentration level (0.1ng/mL), against 76% for UHPSFC-MS/MS and only 14% for GC-MS/MS. Finally, the weakest matrix effects were obtained with UHPSFC-MS/MS with 76% of the analytes displaying relative matrix effect between -20 and 20%, while the GC-MS/MS reference method displayed very strong matrix effects (over 100%) for all of the anabolic agents.

  13. Liquid chromatography and supercritical fluid chromatography as alternative techniques to gas chromatography for the rapid screening of anabolic agents in urine.

    PubMed

    Desfontaine, Vincent; Nováková, Lucie; Ponzetto, Federico; Nicoli, Raul; Saugy, Martial; Veuthey, Jean-Luc; Guillarme, Davy

    2016-06-17

    This work describes the development of two methods involving supported liquid extraction (SLE) sample treatment followed by ultra-high performance liquid chromatography or ultra-high performance supercritical fluid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS and UHPSFC-MS/MS) for the screening of 43 anabolic agents in human urine. After evaluating different stationary phases, a polar-embedded C18 and a diol columns were selected for UHPLC-MS/MS and UHPSFC-MS/MS, respectively. Sample preparation, mobile phases and MS conditions were also finely tuned to achieve highest selectivity, chromatographic resolution and sensitivity. Then, the performance of these two methods was compared to the reference routine procedure for steroid analyses in anti-doping laboratories, which combines liquid-liquid extraction (LLE) followed by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS). For this purpose, urine samples spiked with the compounds of interest at five different concentrations were analyzed using the three analytical platforms. The retention and selectivity of the three techniques were very different, ensuring a good complementarity. However, the two new methods displayed numerous advantages. The overall procedure was much faster thanks to high throughput SLE sample treatment using 48-well plates and faster chromatographic analysis. Moreover, the highest sensitivity was attained using UHPLC-MS/MS with 98% of the doping agents detected at the lowest concentration level (0.1ng/mL), against 76% for UHPSFC-MS/MS and only 14% for GC-MS/MS. Finally, the weakest matrix effects were obtained with UHPSFC-MS/MS with 76% of the analytes displaying relative matrix effect between -20 and 20%, while the GC-MS/MS reference method displayed very strong matrix effects (over 100%) for all of the anabolic agents. PMID:27185056

  14. The analysis of aqueous mixtures using liquid chromatography-electrospray mass spectrometry

    SciTech Connect

    Johnson, S.

    1999-02-12

    The focus of this dissertation is the use of chromatographic methods coupled with electrospray mass spectrometry (ES-MS) for the determination of both organic and inorganic compounds in aqueous solutions. The combination of liquid chromatography (LC) methods and ES-MS offers one of the foremost methods for determining compounds in complex aqueous solutions. In this work, LC-ES-MS methods are devised using ion exclusion chromatography, reversed phase chromatography, and ion exchange chromatography, as well as capillary electrophoresis (CE). For an aqueous sample, these LC-ES-MS and CE-ES-MS techniques require no sample preparation or analyte derivatization, which makes it possible to observe a wide variety of analytes as they exist in solution. The majority of this work focuses on the use of LC-ES-MS for the determination of unknown products and intermediates formed during electrochemical incineration (ECI), an experimental waste remediation process. This report contains a general introduction to the project and the general conclusions. Four chapters have been removed for separate processing. Titles are: Chapter 2: Determination of small carboxylic acids by ion exclusion chromatography with electrospray mass spectrometry; Chapter 3: Electrochemical incineration of benzoquinone in aqueous media using a quaternary metal oxide electrode in the absence of a soluble supporting electrolyte; Chapter 4: The determination of electrochemical incineration products of 4-chlorophenol by liquid chromatography-electrospray mass spectrometry; and Chapter 5: Determination of small carboxylic acids by capillary electrophoresis with electrospray mass spectrometry.

  15. 3D printed metal columns for capillary liquid chromatography.

    PubMed

    Sandron, S; Heery, B; Gupta, V; Collins, D A; Nesterenko, E P; Nesterenko, P N; Talebi, M; Beirne, S; Thompson, F; Wallace, G G; Brabazon, D; Regan, F; Paull, B

    2014-12-21

    Coiled planar capillary chromatography columns (0.9 mm I.D. × 60 cm L) were 3D printed in stainless steel (316L), and titanium (Ti-6Al-4V) alloys (external dimensions of ~5 × 30 × 58 mm), and either slurry packed with various sized reversed-phase octadecylsilica particles, or filled with an in situ prepared methacrylate based monolith. Coiled printed columns were coupled directly with 30 × 30 mm Peltier thermoelectric direct contact heater/cooler modules. Preliminary results show the potential of using such 3D printed columns in future portable chromatographic devices. PMID:25285334

  16. Gold nanoparticle decorated graphene oxide/silica composite stationary phase for high-performance liquid chromatography.

    PubMed

    Liang, Xiaojing; Wang, Xusheng; Ren, Haixia; Jiang, Shengxiang; Wang, Licheng; Liu, Shujuan

    2014-06-01

    In the initial phase of this study, graphene oxide (GO)/silica was fabricated by assembling GO onto the silica particles, and then gold nanoparticles (GNPs) were used to modify the GO/silica to prepare a novel stationary phase for high-performance liquid chromatography. The new stationary phase could be used in both reversed-phase chromatography and hydrophilic interaction liquid chromatography modes. Good separations of alkylbenzenes, isomerides, amino acids, nucleosides, and nucleobases were achieved in both modes. Compared with the GO/silica phase and GNPs/silica phase, it is found that except for hydrophilicity, large π-electron systems, hydrophobicity, and coordination functions, this new stationary phase also exhibited special separation performance due to the combination of 2D GO with zero-dimensional GNPs. PMID:24723561

  17. Gold nanoparticle decorated graphene oxide/silica composite stationary phase for high-performance liquid chromatography.

    PubMed

    Liang, Xiaojing; Wang, Xusheng; Ren, Haixia; Jiang, Shengxiang; Wang, Licheng; Liu, Shujuan

    2014-06-01

    In the initial phase of this study, graphene oxide (GO)/silica was fabricated by assembling GO onto the silica particles, and then gold nanoparticles (GNPs) were used to modify the GO/silica to prepare a novel stationary phase for high-performance liquid chromatography. The new stationary phase could be used in both reversed-phase chromatography and hydrophilic interaction liquid chromatography modes. Good separations of alkylbenzenes, isomerides, amino acids, nucleosides, and nucleobases were achieved in both modes. Compared with the GO/silica phase and GNPs/silica phase, it is found that except for hydrophilicity, large π-electron systems, hydrophobicity, and coordination functions, this new stationary phase also exhibited special separation performance due to the combination of 2D GO with zero-dimensional GNPs.

  18. [Determination of five triazine herbicides in infant milk powder by high performance liquid chromatography coupled with ionic liquid-based homogeneous liquid-liquid microextraction].

    PubMed

    Zhang, Liyuan; Yao, Di; Li, Na; Zhang, Hanqi; Yu, Aimin

    2015-07-01

    A high performance liquid chromatography coupled with homogeneous liquid-liquid microextraction was developed for the determination of five triazine herbicides in infant milk powders. The ionic liquid was used as microextraction solvent. The separation of the herbicides was performed on an Eclipse XDB-C18 column using acetonitrile and water as mobile phases in gradient mode. The effects of homogeneous liquid-liquid extraction conditions on the experimental results were investigated in detail. Under the optimized experimental conditions, the calibration curves for determining the analytes were linear and the correlation coefficients were ≥ 0.9992. The limits of detection for cyanazine, desmetryn, terbumeton, terbuthylazine and dimethametryn were 12.1, 13.8, 11.8, 14.6 and 13.7 μg/kg, respectively. The recoveries of the analytes spiked in four infant milk powders ranged from 92.2% to 103.2% and the relative standard deviations were lower than 6%. This method is sensitive, simple, and suitable for the determination of triazine herbicides in milk powder samples. PMID:26672205

  19. [Determination of five triazine herbicides in infant milk powder by high performance liquid chromatography coupled with ionic liquid-based homogeneous liquid-liquid microextraction].

    PubMed

    Zhang, Liyuan; Yao, Di; Li, Na; Zhang, Hanqi; Yu, Aimin

    2015-07-01

    A high performance liquid chromatography coupled with homogeneous liquid-liquid microextraction was developed for the determination of five triazine herbicides in infant milk powders. The ionic liquid was used as microextraction solvent. The separation of the herbicides was performed on an Eclipse XDB-C18 column using acetonitrile and water as mobile phases in gradient mode. The effects of homogeneous liquid-liquid extraction conditions on the experimental results were investigated in detail. Under the optimized experimental conditions, the calibration curves for determining the analytes were linear and the correlation coefficients were ≥ 0.9992. The limits of detection for cyanazine, desmetryn, terbumeton, terbuthylazine and dimethametryn were 12.1, 13.8, 11.8, 14.6 and 13.7 μg/kg, respectively. The recoveries of the analytes spiked in four infant milk powders ranged from 92.2% to 103.2% and the relative standard deviations were lower than 6%. This method is sensitive, simple, and suitable for the determination of triazine herbicides in milk powder samples.

  20. Microwave-assisted ionic liquid homogeneous liquid-liquid microextraction coupled with high performance liquid chromatography for the determination of anthraquinones in Rheum palmatum L.

    PubMed

    Wang, Zhibing; Hu, Jianxue; Du, Hongxia; He, Shuang; Li, Qing; Zhang, Hanqi

    2016-06-01

    The microwave-assisted ionic liquid homogeneous liquid-liquid microextraction (MA-IL-HLLME) coupled with high performance liquid chromatography with diode array detection (HPLC-DAD) was developed for the determination of anthraquinones, including aloe-emodin, emodin, chrysophanol and physcion in root of Rheum palmatum L. Several experimental parameters influencing the extraction efficiency, including amount of sample, type and volume of ionic liquid, volume and pH value of extraction medium, microwave power and extraction time, concentration of NH4PF6 as well as centrifugal condition were optimized. When 140μL of ionic liquid ([C8MIM][BF4]) was used as an extraction solvent, target analytes can be extracted from sample matrix in one minute with the help of microwave irradiation. The MA-IL-HLLME is simple and quick. The calibration curves exhibited good linear relationship (r>0.9984). The limits of detection and quantification were in the range of 0.015-0.026 and 0.051-0.088μgmL(-1), respectively. The spiked recovery for each analyte was in the range of 81.13-93.07% with relative standard deviations lower than 6.89%. The present method is free of volatile organic solvents, and represents lower expenditures of sample, extraction time and solvent, compared with ultrasonic and heat reflux extraction. The results indicated that the present method can be successfully applied to the determination of anthraquinones in medicinal plant.

  1. Determination of low molecular weight thiols using monobromobimane fluorescent labeling and high-performance liquid chromatography

    NASA Technical Reports Server (NTRS)

    Fahey, Robert C.; Newton, Gerald L.

    1988-01-01

    Methods are described for the preparation and high-performance liquid chromatography (HPLC) analysis of monobromobimane derivatives of low molecular weight thiols in extracts of biological samples. Typical problems encountered in the development and application of these methods are discussed. Analysis of mung bean extract is used as an example.

  2. Determination of febuxostat in human plasma using ultra-performance liquid chromatography tandem mass spectrometry.

    PubMed

    Lukram, Ojikumar; Parmar, Shivaji; Hande, Amit

    2013-06-01

    A rapid and sensitive liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of febuxostat in human plasma. The liquid-liquid extraction technique was used for the extraction of febuxostat from human plasma using trandolapril as the internal standard (IS). Chromatography was performed on a ultra-performance liquid chromatography (UPLC) BEH C18, 50 mm X 2.1 mm, 1.7 µm particle size column, with the mobile phase consisting of 0.1% formic acid and acetonitrile (in a 25:75 ratio), followed by detection using mass spectrometry. The method involves a simple reversed isocratic chromatography condition and mass spectrometry detection, which enables detection at sub-microgram levels. The method was validated and the lower limit of quantification for febuxostat was found to be 0.075 µg/ml. The mean recovery for febuxostat ranged from 100.9 to 106.5%. This method increased the sensitivity and selectivity; resulting in high-throughput analysis of febuxostat using commercially available IS for pharmacokinetic, bioavailability, and bioequivalence studies, with a chromatographic run time of 1.5 min only.

  3. An Advanced, Interactive, High-Performance Liquid Chromatography Simulator and Instructor Resources

    ERIC Educational Resources Information Center

    Boswell, Paul G.; Stoll, Dwight R.; Carr, Peter W.; Nagel, Megan L.; Vitha, Mark F.; Mabbott, Gary A.

    2013-01-01

    High-performance liquid chromatography (HPLC) simulation software has long been recognized as an effective educational tool, yet many of the existing HPLC simulators are either too expensive, outdated, or lack many important features necessary to make them widely useful for educational purposes. Here, a free, open-source HPLC simulator is…

  4. Benzoin Condensation: Monitoring a Chemical Reaction by High-Pressure Liquid Chromatography

    ERIC Educational Resources Information Center

    Bhattacharya, Apurba; Purohit, Vikram C.; Bellar, Nicholas R.

    2004-01-01

    High-pressure liquid chromatography (HPLC) is the preferred method of separating a variety of materials in complex mixtures such as pharmaceuticals, polymers, soils, food products and biological fluids and is also considered to be a powerful analytical tool in both academia and industry. The use of HPLC analysis as a means of monitoring and…

  5. Analysis of Currently Available Analgesic Tablets by Modern Liquid Chromatography: An Undergraduate Laboratory Introduction to HPLC.

    ERIC Educational Resources Information Center

    Kagel, R. A.; Farwell, S. O.

    1983-01-01

    Background information, procedures, and results, are provided for an undergraduate experiment in which analgesic tablets are analyzed using liquid chromatography. The experiment, an improved, modified version of the Waters Associates Inc. experiment, is simple to prepare, requiring little glassware and minimal sample manipulation by students. (JN)

  6. Extraction and Purification of Glucoraphanin by Preparative High-Performance Liquid Chromatography (HPLC)

    ERIC Educational Resources Information Center

    Lee, Iris; Boyce, Mary C.

    2011-01-01

    A student activity that focuses on the isolation of glucoraphanin from broccoli using preparative high-performance liquid chromatography (HPLC) is presented here. Glucoraphanin is a glucosinolate, whose byproducts are known to possess anticancer properties. It is present naturally at high levels in broccoli and other "Brassica" vegetables. This…

  7. The Separation and Identification of Straight Chain Hydrocarbons: An Experiment Using Gas-Liquid Chromatography.

    ERIC Educational Resources Information Center

    Benson, G. A.

    1982-01-01

    An experiment using gas-liquid chromatography is discussed, introducing the student to concept of dead volume and its measurement, idea and use of an internal reference compound, and to linear relationship existing between measurements of a separation on two different stationary phases. (Author/SK)

  8. (PRESENT AT NCCU) ANALYSIS OF SELECTED PYRETHROID PESTICIDES USING REVERSE PHASE HIGH LIQUID CHROMATOGRAPHY

    EPA Science Inventory

    This research was conducted in cooperation with EPA Region 4 in Athens, GA to develop a method to analyze selected pyrethroid pesticides using Reverse Phase-High Pressure Liquid Chromatography (HPLC). This HPLC method will aid researchers in separating and identifying these pyre...

  9. ANALYSIS OF SELECTED PYRETHROID PESTICIDES USING REVERSE PHASE HIGH PRESSURE LIQUID CHROMATOGRAPHY/UV

    EPA Science Inventory

    This research was conducted in cooperation with EPA Region 4 in Athens, GA to develop a method to analyze selected pyrethroid pesticides using Reverse Phase-High Pressure Liquid Chromatography (HPLC). This HPLC method will aid researchers in separating and identifying these py...

  10. Sensitive, Preclinical Detection of Prions in Brain by nanospray liquid chromatography/tandem mass spectrometry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    More sensitive detection of prions in brain is important because it would allow early detection of disease in young animals and assure a safer food supply. We quantitated the amount of proteinase K-resistant prion protein (PrP 27-30) by use of nano-scale liquid chromatography coupled to a tandem ma...

  11. High-Performance Liquid Chromatography in the Undergraduate Chemical Engineering Laboratory

    ERIC Educational Resources Information Center

    Frey, Douglas D.; Guo, Hui; Karnik, Nikhila

    2013-01-01

    This article describes the assembly of a simple, low-cost, high-performance liquid chromatography (HPLC) system and its use in the undergraduate chemical engineering laboratory course to perform simple experiments. By interpreting the results from these experiments students are able to gain significant experience in the general method of…

  12. High pressure liquid chromatography of cannabinoids as their fluorescent dansyl derivatives.

    PubMed

    Abbott, S R; Abu-Shumays, A

    1975-01-01

    Four different cannabinoids were converted to their 1-dimethyl-aminonaphthalene-5-sulfonate (dansyl) derivatives. Standards mixtures were separated by high pressure liquid chromatography and detected with an experimental filter fluorometer with sub-nanogram sensitivity. The effect of solvent properties on fluorescence spectra and quantum yields was also studied.

  13. DETERMINATION OF CHLOROPHENOLS, NITROPHENOLS, AND METHYLPHENOLS IN GROUND-WATER SAMPLES USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    EPA Science Inventory

    A high performance liquid chromatography (HPLC) method was developed to quantitatively determine phenolic compounds and their isomers in aqueous samples. The HPLC method can analyze a mixture of 15 contaminants in the same analytical run with an analysis time of 25 minutes. The...

  14. DETERMINATION OF CHLOROPHEONIS, NITROPHENOIS AND METHYLPHENOIS IN GROUND-WATER SAMPLES USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    EPA Science Inventory

    A high performance liquid chromatography (HPLC) method was developed to quantitatively determine phenolic compounds and their isomers in aqueous samples. The HPLC method can analyze a mixture of 15 contaminants in the same analytical run with an analysis time of 25 minutes. The...

  15. MEASUREMENT OF OXIDATIVE STRESS PARAMETERS USING LIQUID CHROMATOGRAPHY - TANDEM MASS SPECTROSCOPY (LC-MS/MS)

    EPA Science Inventory

    What is the study?
    An invited review article. Measurement of oxidative stress parameters using liquid chromatography-tandem mass spectroscopy (LC-MS/MS)
    Why was it done?
    Although oxidative stress is frequently cited as a cause of various adverse biological eff...

  16. Determination of cyclamate by high-performance liquid chromatography with indirect photometry.

    PubMed

    Herrmann, A; Damawandi, E; Wagmann, M

    1983-12-01

    Most artificial sweeteners have been determined by high-performance liquid chromatography, but not cyclamate. We propose a simple method using standard equipment without any chemical reaction involving the cyclamate ion by applying the technique of indirect photometry. Saccharin, dulcin and aspartame may also be determined, if present.

  17. Sugar Determination in Foods with a Radially Compressed High Performance Liquid Chromatography Column.

    ERIC Educational Resources Information Center

    Ondrus, Martin G.; And Others

    1983-01-01

    Advocates use of Waters Associates Radial Compression Separation System for high performance liquid chromatography. Discusses instrumentation and reagents, outlining procedure for analyzing various foods and discussing typical student data. Points out potential problems due to impurities and pump seal life. Suggests use of ribose as internal…

  18. Going Beyond, Going Further: An Inexpensive Experiment for the Introduction of High Performance Liquid Chromatography.

    ERIC Educational Resources Information Center

    Bidlingmeyer, Brian A.; Warren, F. Vincent, Jr.

    1984-01-01

    Background information, materials needed, laboratory procedures, and typical results are provided for five high performance liquid chromatography experiments (three isocratic and two step gradient separations). Suggestions for further experimentation are also provided, including quantitative determinations and separation of charged solutes. (JN)

  19. Quantitative separation of tetralin hydroperoxide from its decomposition products by high performance liquid chromatography

    NASA Technical Reports Server (NTRS)

    Worstell, J. H.; Daniel, S. R.

    1981-01-01

    A method for the separation and analysis of tetralin hydroperoxide and its decomposition products by high pressure liquid chromatography has been developed. Elution with a single, mixed solvent from a micron-Porasil column was employed. Constant response factors (internal standard method) over large concentration ranges and reproducible retention parameters are reported.

  20. High Performance Liquid Chromatography of Some Analgesic Compounds: An Instrumental Analysis Experiment.

    ERIC Educational Resources Information Center

    Haddad, Paul; And Others

    1983-01-01

    Background information, procedures, and results are provided for an experiment demonstrating techniques of solvent selection, gradient elution, pH control, and ion-pairing in the analysis of an analgesic mixture using reversed-phase liquid chromatography on an octadecylsilane column. Although developed using sophisticated/expensive equipment, less…

  1. Detection of Geothermal Phosphite Using High Performance Liquid Chromatography

    PubMed Central

    Pech, Herbe; Henry, Amanda; Khachikian, Crist S.; Salmassi, Tina M.; Hanrahan, Grady; Foster, Krishna L.

    2009-01-01

    Little is known about the pre-biotic mechanisms that initiated the bioavailability of phosphorus, an element essential to life. A better understanding of phosphorus speciation in modern earth environments representative of early earth, may help to elucidate the origins of bioavailable phosphorus. This paper presents the first quantitative measurements of phosphite in a pristine geothermal pool representative of early earth. Phosphite and phosphate were initially identified and quantified in geothermal pool and stream samples at Hot Creek Gorge near Mammoth Lakes, California using suppressed conductivity ion chromatography. Results confirmed the presence of 0.06 ± 0.02 μM of phosphite and 0.05 ± 0.01 μM of phosphate in a geothermal pool. In the stream, phosphite concentrations were below detection limit (0.04 μM) and phosphate was measured at 1.06 ± 0.36 μM. The presence of phosphite in the geothermal pool was confirmed using both chemical oxidation and ion chromatography/mass spectrometry. PMID:19921877

  2. Detection of geothermal phosphite using high-performance liquid chromatography.

    PubMed

    Pech, Herbe; Henry, Amanda; Khachikian, Crist S; Salmassi, Tina M; Hanrahan, Grady; Foster, Krishna L

    2009-10-15

    Little is known about the prebiotic mechanisms that initiated the bioavailability of phosphorus, an element essential to life. A better understanding of phosphorus speciation in modern earth environments representative of early earth may help to elucidate the origins of bioavailable phosphorus. This paper presents the first quantitative measurements of phosphite in a pristine geothermal pool representative of early earth. Phosphite and phosphate were initially identified and quantified in geothermal pool and stream samples at Hot Creek Gorge near Mammoth Lakes, California, using suppressed conductivity ion chromatography. Results confirmed the presence of 0.06 +/- 0.02 microM of phosphite and 0.05 +/- 0.01 microM of phosphate in a geothermal pool. In the stream, phosphite concentrations were below detection limit (0.04 microM) and phosphate was measured at 1.06 +/- 0.36 microM. The presence of phosphite in the geothermal pool was confirmed using both chemical oxidation and ion chromatography/mass spectrometry.

  3. Characterization of goat colostrum oligosaccharides by nano-liquid chromatography on chip quadrupole time-of-flight mass spectrometry and hydrophilic interaction liquid chromatography-quadrupole mass spectrometry.

    PubMed

    Martín-Ortiz, A; Salcedo, J; Barile, D; Bunyatratchata, A; Moreno, F J; Martin-García, I; Clemente, A; Sanz, M L; Ruiz-Matute, A I

    2016-01-01

    A detailed qualitative and quantitative characterization of goat colostrum oligosaccharides (GCO) has been carried out for the first time. Defatted and deproteinized colostrum samples, previously treated by size exclusion chromatography (SEC) to remove lactose, were analyzed by nanoflow liquid chromatography-quadrupole-time of flight mass spectrometry (Nano-LC-Chip-Q-TOF MS). Up to 78 oligosaccharides containing hexose, hexosamine, fucose, N-acetylneuraminic acid or N-glycolylneuraminic acid monomeric units were identified in the samples, some of them detected for the first time in goat colostra. As a second step, a hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-MS) methodology was developed for the separation and quantitation of the main GCO, both acidic and neutral carbohydrates. Among other experimental chromatographic conditions, mobile phase additives and column temperature were evaluated in terms of retention time, resolution, peak width and symmetry of target carbohydrates. Narrow peaks (wh: 0.2-0.6min) and good symmetry (As: 0.8-1.4) were obtained for GCO using an acetonitrile:water gradient with 0.1% ammonium hydroxide at 40°C. These conditions were selected to quantify the main oligosaccharides in goat colostrum samples. Values ranging from 140 to 315mgL(-1) for neutral oligosaccharides and from 83 to 251mgL(-1) for acidic oligosaccharides were found. The combination of both techniques resulted to be useful to achieve a comprehensive characterization of GCO.

  4. Characterization of goat colostrum oligosaccharides by nano-liquid chromatography on chip quadrupole time-of-flight mass spectrometry and hydrophilic interaction liquid chromatography-quadrupole mass spectrometry

    PubMed Central

    Martín-Ortiz, A.; Salcedo, J.; Barile, D.; Bunyatratchata, A.; Moreno, F.J.; Martin-García, I.; Clemente, A.; Sanz, M.L.; Ruiz-Matute, A.I.

    2016-01-01

    A detailed qualitative and quantitative characterization of goat colostrum oligosaccharides (GCO) has been carried out for the first time. Defatted and deproteinized colostrum samples, previously treated by size exclusion chromatography (SEC) to remove lactose, were analyzed by nanoflow liquid chromatography-quadrupole-time of flight mass spectrometry (Nano-LC-Chip-Q-TOF MS). Up to 78 oligosaccharides containing hexose, hexosamine, fucose, N-acetylneuraminic acid or N-glycolylneuraminic acid monomeric units were identified in the samples, some of them detected for the first time in goat colostra. As a second step, a hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-MS) methodology was developed for the separation and quantitation of the main GCO, both acidic and neutral carbohydrates. Among other experimental chromatographic conditions, mobile phase additives and column temperature were evaluated in terms of retention time, resolution, peak width and symmetry of target carbohydrates. Narrow peaks (wh: 0.2–0.6 min) and good symmetry (As: 0.8–1.4) were obtained for GCO using an acetonitrile:water gradient with 0.1% ammonium hydroxide at 40 °C. These conditions were selected to quantify the main oligosaccharides in goat colostrum samples. Values ranging from 140 to 315 mg L−1 for neutral oligosaccharides and from 83 to 251 mg L−1 for acidic oligosaccharides were found. The combination of both techniques resulted to be useful to achieve a comprehensive characterization of GCO. PMID:26427327

  5. Extraction and determination of chloramphenicol in feed water, milk, and honey samples using an ionic liquid/sodium citrate aqueous two-phase system coupled with high-performance liquid chromatography.

    PubMed

    Han, Juan; Wang, Yun; Yu, Cui-lan; Yan, Yong-sheng; Xie, Xue-qiao

    2011-01-01

    A green, simple, non-toxic, and sensitive sample pretreatment procedure coupled with high-performance liquid chromatography (HPLC) was developed for the analysis of chloramphenicol (CAP) that exploits an aqueous two-phase system based on imidazolium ionic liquid (1-butyl-3-methylimidazolium tetrafluoroborate, [Bmim]BF(4)) and organic salt (Na(3)C(6)H(5)O(7)) using a liquid-liquid extraction technique. The influence factors on partition behaviors of CAP were studied, including the type and amount of salts, the pH value, the volume of [Bmim]BF(4), and the extraction temperature. Extraction efficiency of the CAP was found to increase with increasing temperature and the volume of [Bmim]BF(4). Thermodynamic studies indicated that hydrophobic interactions were the main driving force, although electrostatic interactions and salting-out effects were also important for the transfer of the CAP. Under the optimal conditions, 90.1% of the CAP could be extracted into the ionic liquid-rich phase in a single-step extraction. This method was practical when applied to the analysis of CAP in feed water, milk, and honey samples with a linear range of 2~1,000 ng mL(-1). The method yielded a limit of detection of 0.3 ng mL(-1) and a limit of quantification of 1.0 ng mL(-1). The recovery of CAP was 90.4-102.7% from aqueous samples of real feed water, milk, and honey samples by the proposed method. This novel process is much simpler and more environmentally friendly and is suggested to have important applications for the separation of antibiotics. PMID:21063686

  6. Position sensitive radioactivity detection for gas and liquid chromatography

    DOEpatents

    Cochran, Joseph L.; McCarthy, John F.; Palumbo, Anthony V.; Phelps, Tommy J.

    2001-01-01

    A method and apparatus are provided for the position sensitive detection of radioactivity in a fluid stream, particularly in the effluent fluid stream from a gas or liquid chromatographic instrument. The invention represents a significant advance in efficiency and cost reduction compared with current efforts.

  7. Determination of Atmospheric Hydroxyl Radical by Liquid Phase Scrubbing and High Performance Liquid Chromatography.

    NASA Astrophysics Data System (ADS)

    Chen, Xiaohui

    A new in situ method for determining atmospheric hydroxyl radical (OH) was developed. This method is based on liquid phase scrubbing technique and high performance liquid chromatography (LPS-HPLC). The sampling system employs glass bubblers to trap atmospheric OH into a buffered solution containing the chemical probe, salicylic acid (o-hydroxybenzoic acid, OHBA). The reaction of OHBA with OH produces a stable fluorescent product, 2,5-dihydroxybenzoic acid (2,5-DHBA), which is determined by reverse phase HPLC and fluorescence detection. The atmospheric OH concentration ( (OH)) is inferred from the resulting 2,5-DHBA amount in the aqueous scrubbing solution, fraction of 2,5-DHBA in the products, air sampling flow rate, sampling time, local pressure and temperature, etc. HPLC separation efficiency and fluorescence detection sensitivity for 2,5-DHBA have been studied. The results indicate that: the reagent blank can be controlled by suitable recrystallization; pH affects both separation and detection processes; the fluorometer should be adjusted to reach its highest signal-to-noise ratio by light source selection, flow cell size selection, wavelength selection, etc. Preliminary column switch experiments reveal the possibility to automate the whole sampling and detection system to enhance the temporal resolution. During an intercomparison of tropospheric OH measurement techniques at the Caribou site, CO (relatively unpolluted air) in Fall 1993, overlapping data were obtained with long path absorption and ion-assisted coupled with MS methods. LPS -HPLC day-time (OH) s, which range from {< }10^6 to 6times10 ^6 radicals/cm^3, agree well with those derived from the other two methods, especially the latter. LPS-HPLC (OH) depends linearly on the combined effects of solar flux, ozone and water vapor, however, it has a nonlinear dependence on NO _{x} and hydrocarbons. These results are consistent with that predicted from photochemical models. Experimental results and model

  8. Hyphenated and comprehensive liquid chromatography × gas chromatography-mass spectrometry for the identification of Mycobacterium tuberculosis.

    PubMed

    Mourão, Marta P B; Denekamp, Ilse; Kuijper, Sjoukje; Kolk, Arend H J; Janssen, Hans-Gerd

    2016-03-25

    Tuberculosis is one of the world's most emerging public health problems, particularly in developing countries. Chromatography based methods have been used to tackle this epidemic by focusing on biomarker detection. Unfortunately, interferences from lipids in the sputum matrix, particularly cholesterol, adversely affect the identification and detection of the marker compounds. The present contribution describes the serial combination of normal phase liquid chromatography (NPLC) with thermally assisted hydrolysis and methylation followed by gas chromatography-mass spectrometry (THM-GC-MS) to overcome the difficulties of biomarker evaluation. The in-series combination consists of an LC analysis where fractions are collected and then transferred to the THM-GC-MS system. This was either done with comprehensive coupling, transferring all the fractions, or with hyphenated interfacing, i.e. off-line multi heart-cutting, transferring only selected fractions. Owing to the high sensitivity and selectivity of LC as a sample pre-treatment method, and to the high specificity of the MS as a detector, this analytical approach, NPLC × THM-GC-MS, is extremely sensitive. The results obtained indicate that this analytical set-up is able to detect down to 1 × 10(3) mycobacteria/mL of Mycobacterium tuberculosis strain 124, spiked in blank sputum samples. It is a powerful analytical tool and also has great potential for full automation. If further studies demonstrate its usefulness when applied blind in real sputum specimens, this technique could compete with the current smear microscopy in the early diagnosis of tuberculosis.

  9. Determination of synthetic hormones in animal urine by high-performance liquid chromatography/mass spectrometry.

    PubMed

    Rúbies, Antoni; Cabrera, Albert; Centrich, Francesc

    2007-01-01

    A method was developed for the extraction of stanozolol, taleranol, zeranol, hexestrol, dienestrol, ethynylestradiol, diethylstilbestrol, and trenbolone from animal urine. The analytes were extracted from the matrix by enzymatic hydrolysis, solid-phase extraction, and liquid-liquid extraction. Detection and quantitation were performed on a high-performance liquid chromatography system coupled to a triple quadrupole mass spectrometer. The identification criteria met the European Union regulations. Validation of this method established a decision limit between 0.2 and 0.9 microg/L and a detection capability between 0.3 and 1.0 microg/L, depending on the analyte.

  10. Determination of synthetic hormones in animal urine by high-performance liquid chromatography/mass spectrometry.

    PubMed

    Rúbies, Antoni; Cabrera, Albert; Centrich, Francesc

    2007-01-01

    A method was developed for the extraction of stanozolol, taleranol, zeranol, hexestrol, dienestrol, ethynylestradiol, diethylstilbestrol, and trenbolone from animal urine. The analytes were extracted from the matrix by enzymatic hydrolysis, solid-phase extraction, and liquid-liquid extraction. Detection and quantitation were performed on a high-performance liquid chromatography system coupled to a triple quadrupole mass spectrometer. The identification criteria met the European Union regulations. Validation of this method established a decision limit between 0.2 and 0.9 microg/L and a detection capability between 0.3 and 1.0 microg/L, depending on the analyte. PMID:17474533

  11. Reversed-phase High Performance Liquid Chromatography of proteins.

    PubMed

    Josic, Djuro; Kovac, Spomenka

    2010-08-01

    Reversed-phase HPLC (RP-HPLC) is one of most important techniques for protein separations and the method of choice for peptide separation. RP-HPLC has been applied on the nano, micro, and analytical scale, and has also been scaled up for preparative purifications, to large industrial scale. Because of its compatibility with mass spectrometry, RP-HPLC is an indispensable tool in proteomic research. With modern instrumentation and columns, complex mixtures of peptides and proteins can be separated at attomolar levels for further analysis. In addition, preparative RP-HPLC is often used for large-scale purification of proteins. This unit provides protocols for packing and testing a column, protein separation by use of gradient or step elution, desalting of protein solutions, and separation of enzymatic digests before mass spectrometric analyses. A protocol is also provided for cleaning, regenerating, and storing reversed-phase chromatography columns.

  12. Separation of carbohydrates using hydrophilic interaction liquid chromatography.

    PubMed

    Fu, Qing; Liang, Tu; Li, Zhenyu; Xu, Xiaoyong; Ke, Yanxiong; Jin, Yu; Liang, Xinmiao

    2013-09-20

    A strategy was developed to rapidly evaluate chromatographic properties of hydrophilic interaction chromatography (HILIC) columns for separating carbohydrates. Seven HILIC columns (Silica, Diol, TSK Amide-80, XAmide, Click Maltose, Click β-CD, and Click TE-Cys columns) were evaluated by using three monosaccharide and seven disaccharides as probes. The influence of column temperature on the peak shape and tautomerization of carbohydrates, as well as column selectivity were investigated. The influence of surface charge property on the retention was also studied by using glucose, glucuronic acid, and glucosamine, which indicated that buffer salt concentration and pH value in mobile phase was necessary to control the ionic interactions between ionic carbohydrates and HILIC columns. According to evaluation results, the XAmide column was selected as an example to establish experimental schemes for separation of complex mixtures of oligosaccharide.

  13. Single-Step Ironmaking from Ore to Improve Energy Efficiency

    SciTech Connect

    S.K. Kawatra; B. Anamerie; T.C. Eisele

    2005-10-01

    The pig iron nugget process was developed as an alternative to the traditional blast furnace process by Kobe Steel. The process aimed to produce pig iron nuggets, which have similar chemical and physical properties to blast furnace pig iron, in a single step. The pig iron nugget process utilizes coal instead of coke and self reducing and fluxing dried green balls instead of pellets and sinters. In this process the environmental emissions caused by coke and sinter production, and energy lost between pellet induration (heat hardening) and transportation to the blast furnace can be eliminated. The objectives of this research were to (1) produce pig iron nuggets in the laboratory, (2) characterize the pig iron nugget produced and compare them with blast furnace pig iron, (3) investigate the furnace temperature and residence time effects on the pig iron nugget production, and (4) optimize the operational furnace temperatures and residence times. The experiments involved heat treatment of self reducing and fluxing dried green balls at various furnace temperatures and residence times. Three chemically and physically different products were produced after the compete reduction of iron oxides to iron depending on the operational furnace temperatures and/or residence times. These products were direct reduced iron (DRI), transition direct reduced iron (TDRI), and pig iron nuggets. The increase in the carbon content of the system as a function of furnace temperature and/or residence time dictated the formation of these products. The direct reduced iron, transition direct reduced iron, and pig iron nuggets produced were analyzed for their chemical composition, degree of metallization, apparent density, microstructure and microhardness. In addition, the change in the carbon content of the system with the changing furnace temperature and/or residence time was detected by optical microscopy and Microhardness measurements. The sufficient carbon dissolution required for the

  14. Characterization of sodium carboxymethyl cellulose by comprehensive two-dimensional liquid chromatography.

    PubMed

    Shakun, Maria; Heinze, Thomas; Radke, Wolfgang

    2015-10-01

    Two series of sodium carboxymethyl celluloses (NaCMC) with average degrees of substitution (DS) ranging from 0.45 to 1.55 were synthesized from low molecular mass Avicel cellulose (Avicel samples) and from high molecular mass cotton linters (BWL samples). The samples were characterized by online two-dimensional liquid chromatography using gradient liquid adsorption chromatography in the first and size exclusion chromatography (SEC) in the second dimension. This method allows the simultaneous determination of the chemical composition (DS) and the molar mass distribution within the individual samples. Moreover information was obtained on the dependence of the elution volume in gradient chromatography on molar mass. As expected, evidence for a stronger influence of molar mass on gradient elution volume was found for the low molecular mass NaCMC as compared to the high molecular mass BWL samples. Finally the applicability of the method for the simultaneous separation of blends heterogeneous with respect to chemical composition (DS) and molar mass was demonstrated. Such blends cannot be efficiently separated by either SEC or gradient chromatography alone, nor by simply combining the results of both methods. Only the complete two-dimensional chromatogram can reveal the complexity of such blends, since it reveals the correlations between molar mass and chemical composition.

  15. Determination of oxidized phosphatidylcholines by hydrophilic interaction liquid chromatography coupled to Fourier transform mass spectrometry.

    PubMed

    Sala, Pia; Pötz, Sandra; Brunner, Martina; Trötzmüller, Martin; Fauland, Alexander; Triebl, Alexander; Hartler, Jürgen; Lankmayr, Ernst; Köfeler, Harald C

    2015-04-14

    A novel liquid chromatography-mass spectrometry (LC-MS) approach for analysis of oxidized phosphatidylcholines by an Orbitrap Fourier Transform mass spectrometer in positive electrospray ionization (ESI) coupled to hydrophilic interaction liquid chromatography (HILIC) was developed. This method depends on three selectivity criteria for separation and identification: retention time, exact mass at a resolution of 100,000 and collision induced dissociation (CID) fragment spectra in a linear ion trap. The process of chromatography development showed the best separation properties with a silica-based Kinetex column. This type of chromatography was able to separate all major lipid classes expected in mammalian samples, yielding increased sensitivity of oxidized phosphatidylcholines over reversed phase chromatography. Identification of molecular species was achieved by exact mass on intact molecular ions and CID tandem mass spectra containing characteristic fragments. Due to a lack of commercially available standards, method development was performed with copper induced oxidation products of palmitoyl-arachidonoyl-phosphatidylcholine, which resulted in a plethora of lipid species oxidized at the arachidonoyl moiety. Validation of the method was done with copper oxidized human low-density lipoprotein (LDL) prepared by ultracentrifugation. In these LDL samples we could identify 46 oxidized molecular phosphatidylcholine species out of 99 possible candidates.

  16. Simultaneous determination of seven anticoagulant rodenticides in agricultural products by gel permeation chromatography and liquid chromatography-tandem mass spectrometry.

    PubMed

    Saito-Shida, Shizuka; Nemoto, Satoru; Matsuda, Rieko; Akiyama, Hiroshi

    2016-11-01

    A sensitive and reliable method for the simultaneous determination of hydroxycoumarin-type (brodifacoum, bromadiolone, coumatetralyl, and warfarin) and indandione-type (chlorophacinone, diphacinone, and pindone) rodenticides in agricultural products by gel permeation chromatography (GPC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. The procedure involved extraction of the rodenticides from samples with acetone, followed by liquid-liquid partitioning with hexane/ethyl acetate (1:1, v/v) and 10% sodium chloride aqueous solution, then cleanup using GPC, and finally, analysis using LC-MS/MS. High recoveries from the GPC column were obtained for all rodenticides tested using a mobile phase of acetone/cyclohexane/triethylamine (400:1600:1, v/v/v). An ODS column, which contains low levels of metal impurities, gave satisfactory peak shapes for both hydroxycoumarin- and indandione-type rodenticides in the LC-MS/MS separation. The average recoveries of rodenticides from eight agricultural foods (apple, eggplant, cabbage, orange, potato, tomato, brown rice, and soybean) fortified at 0.0005-0.001 mg/kg ranged from 76 to 116%, except for bromadiolone in orange (53%) and diphacinone in soybean (54%), and the relative standard deviations ranged from 1 to 16%. The proposed method effectively removed interfering components, such as pigments and lipids, and showed high selectivity. In addition, the matrix effects were negligible for most of the rodenticide/food combinations. The results suggest that the proposed method is reliable and suitable for determining hydroxycoumarin- and indandione-type rodenticides in agricultural products. PMID:27428755

  17. Forensic analysis of ignitable liquids in fire debris by comprehensive two-dimensional gas chromatography.

    PubMed

    Frysinger, Glenn S; Gaines, Richard B

    2002-05-01

    The application of comprehensive two-dimensional gas chromatography (GC x GC) for the forensic analysis of ignitable liquids in fire debris is reported. GC x GC is a high resolution, multidimensional gas chromatographic method in which each component of a complex mixture is subjected to two independent chromatographic separations. The high resolving power of GC x GC can separate hundreds of chemical components from a complex fire debris extract. The GC x GC chromatogram is a multicolor plot of two-dimensional retention time and detector signal intensity that is well suited for rapid identification and fingerprinting of ignitable liquids. GC x GC chromatograms were used to identify and classify ignitable liquids, detect minor differences between similar ignitable liquids, track the chemical changes associated with weathering, characterize the chemical composition of fire debris pyrolysates, and detect weathered ignitable liquids against a background of fire debris pyrolysates.

  18. [Determination of dyes in cosmetic by high performance liquid chromatography].

    PubMed

    Wang, J; Hu, J

    1999-09-01

    A reversed-phase high performance liquid chromatographic method was used to detect 5 dyes including p-phenylenediamine in cosmetrics. A Zorbax C8 column was used and mobile phase was V (triethanolamine):V(water):V(acetonitrile) = 0.95:94.05:5 adjusted to pH 7.7 by phosphoric acid. The wavelength 280 nm was selected. Samples were extracted with 95% ethanol by ultrasonic method. The recoveries were 87%-107% and the relative standard deviations were 2.3%-6.4%.

  19. Microporous membrane liquid-liquid extraction coupled on-line with normal-phase liquid chromatography for the determination of cationic surfactants in river and waste water.

    PubMed

    Norberg, J; Thordarson, E; Mathiasson, L; Jönsson, J A

    2000-02-11

    Membrane-based continuous liquid-liquid extraction combined on-line with normal-phase liquid chromatography is proposed for the determination of cationic surfactants in complex aqueous samples. The technique has the potential for complete automation. Selective enrichment of cationic surfactants from spiked river water and waste-water samples with simultaneous removal of matrix constituents, followed by a quantitative transfer of the extract onto a liquid chromatographic column and separation of the surfactant homologues yielding low detection limits, has been realised. The homologues of alkyldimethylbenzylammonium chloride (Dodigen 226) were chosen as model compounds in the method development. Dodigen homologues were ion-paired with heptanoic acid and extracted into chlorobutane by means of microporous membrane liquid-liquid extraction. It was thereby possible to attain an enrichment of over 250 times for one of the homologues, viz. the concentration in the organic liquid is 250 times higher than in the original sample. Detection limits for the three best-detected homologues of the mixture were in the range 0.7-5 microg/l in spiked river water samples. Ion-pair normal-phase liquid chromatography, again with heptanoic acid as counter-ion, gave the necessary separation of the surfactant homologues.

  20. Lattice-fluid model for gas-liquid chromatography.

    PubMed

    Tao, Y; Wells, P S; Yi, X; Yun, K S; Parcher, J F

    1999-11-01

    Lattice-fluid models describe molecular ensembles in terms of the number of lattice sites occupied by molecular species (r-mers) and the interactions between neighboring molecules. The lattice-fluid model proposed by Sanchez and Lacombe (Macromolecules, 1978;11:1145-1156) was used to model specific retention volume data for a series of n-alkane solutes with n-alkane, polystyrene, and poly(dimethylsiloxane) stationary liquid phases. Theoretical equations were derived for the specific retention volume and also for the temperature dependence and limiting (high temperature) values for the specific retention volume. The model was used to predict retention volumes within 10% for the n-alkanes phases; 22% for polystyrene; and from 20 to 70% for PDMS using no adjustable parameters. The temperature derivative (enthalpy) could be calculated within 5% for all of the solutes in nine stationary liquid phases. The limiting value for the specific retention volume at high temperature (entropy controlled state) could be calculated within 10% for all of the systems. The limiting data also provided a new chromatographic method to measure the size parameter, r, for any chromatographic solute using characteristic and size parameters for the stationary phase only. The calculated size parameters of the solutes were consistent, i.e. independent of the stationary phase and agreed within experimental error with the size parameters previously reported from saturated vapor pressure, latent heat of vaporization or density data.

  1. Cigarette filters as adsorbents of solid-phase extraction for determination of fluoroquinolone antibiotics in environmental water samples coupled with high-performance liquid chromatography.

    PubMed

    Chen, Bo; Wang, Weidong; Huang, Yuming

    2012-01-15

    The potential use of cigarette filters (CFs) as solid-phase extraction (SPE) adsorbents for the preconcentration of six fluoroquinolones (FQs) antibacterial agents prior to liquid chromatography was examined in this paper. In order to find a suitable procedure for extraction of the target FQs in one single step, various parameters probably affecting the extraction efficiency including the eluent kind and volume, sample flow rate, pH, ion strength and sample volume were systematically optimized. Under the optimized conditions, the target FQs could be easily extracted by the proposed SPE cartridge. Combination of SPE with HPLC/UV provided detection limits for different FQs of 2-5 ng L(-1) when 500 mL of water sample was processed. The precision of the method, expressed as relative standard deviation, ranged from 4.1 to 6.3% for 2.5 μg L(-1) FQs. The recoveries of FQs spiked in environmental water samples ranged from 76 to 112%. The results obtained from the proposed method demonstrated that CFs-based solid-phase extraction combined with HPLC/UV was suitable for analyzing fluoroquinolones in water samples at ng L(-1) concentration level.

  2. Fast preparation of a highly efficient organic monolith via photo-initiated thiol-ene click polymerization for capillary liquid chromatography.

    PubMed

    Chen, Lianfang; Ou, Junjie; Liu, Zhongshan; Lin, Hui; Wang, Hongwei; Dong, Jing; Zou, Hanfa

    2015-05-15

    A novel organic monolith was firstly prepared in a UV-transparent fused-silica capillary by a single-step approach via photo-initiated thiol-ene click polymerization reaction of 1,2,4-trivinylcyclohexane (TVCH) and pentaerythriol tetra(3-mercaptopropionate) (4SH) within 10min. The effects of both composition of prepolymerization solution and polymerization time on the morphology and permeability of monolithic column were investigated in detail. Then, the optimal condition was acquired to fabricate a homogeneous and permeable organic monolith. The chemical groups of the monolithic column were confirmed by Fourier transform infrared spectroscopy (FT-IR). The SEM graphs showed the organic monolith possessed a uniform porous structure, which promotes the highest column efficiency of ∼133,000 plates per meter for alkylbenzenes at the linear velocity of 0.65mm/s in reversed-phase liquid chromatography. Finally, the organic monolithic column was further applied for separation of basic compounds, pesticides and EPA610, indicating satisfactory separation ability.

  3. Simultaneous determination of neonicotinoid insecticides in human serum and urine using diatomaceous earth-assisted extraction and liquid chromatography-tandem mass spectrometry.

    PubMed

    Yamamuro, Tadashi; Ohta, Hikoto; Aoyama, Mika; Watanabe, Daisuke

    2014-10-15

    A rapid and sensitive analytical method was developed for simultaneous determination of eight neonicotinoid insecticides (acetamiprid, clothianidin, dinotefuran, flonicamid, imidacloprid, nitenpyram, thiacloprid and thiamethoxam) and three specific metabolites of acetamiprid (N-desmethylacetamiprid, 5-(N-acetyl-N-methylaminomethyl)-2-chloropyridine and 5-(N-acetylaminomethyl)-2-chloropyridine) in human serum and urine. A diatomaceous earth-assisted extraction using Extrelut NT3 column with chloroform/2-propanol (3:1, v/v) as eluent was selected for the single step cleanup procedure for all the target compounds. Qualitative and quantitative analyses were conducted by liquid chromatography-tandem mass spectrometry with multiple reaction monitoring mode. The limits of detection and the limits of quantification of eleven compounds were in the ranges of 0.1-0.2ng/mL and 0.5-10ng/mL for serum, 0.1-1ng/mL and 1-10ng/mL for urine, respectively. The extraction recoveries were between 80.9% and 101.8% for serum samples, 91.9% and 106% for urine samples. The intra-day RSDs and the inter-day RSDs were less than 11.5% and 13.2% for serum, less than 8.3% and 8.8% for urine. The proposed procedure will be suitable for forensic investigations of human poisoning cases with neonicotinoid insecticides. This is the first report of simultaneous determination of eight neonicotinoids in serum and urine samples. PMID:25168791

  4. Application of solid-phase extraction and liquid chromatography-mass spectrometry to the determination of neonicotinoid pesticide residues in fruit and vegetables.

    PubMed

    Di Muccio, Alfonso; Fidente, Paola; Barbini, Danilo Attard; Dommarco, Roberto; Seccia, Serenella; Morrica, Patrizia

    2006-03-01

    A rapid and simple method for simultaneous analysis of four neonicotinoid insecticides including acetamiprid, imidacloprid, thiacloprid and thiamethoxam in fruit and vegetable matrices has been developed. For instance, ready-to-use cartridges filled with a macroporous diatomaceous material were used to extract in a single step insecticide residues with dichloromethane from aqueous-acetone extracts of fruits and vegetables. The eluate was evaporated, the residue redissolved with methanol and then analyzed by liquid chromatography-mass spectrometry in the electrospray ionization (ESI) positive mode. Average recoveries of the four pesticides were between 74.5 and 105% at both spiking levels 0.1 and 1.0 mg kg(-1) in peach, pear, courgette, celery and apricot. Relative standard deviations (RSDs) were less than 10% for all of the recovery tests. The calculated limits of quantitation (LOQs) (0.1-0.5 mg kg(-1)) were equal or lower then the maximum residue limits (MRLs) established by European legislation (0.1-0.5 mg kg(-1)). The proposed method is fast, easy to perform and could be utilized for monitoring of pesticides residues. PMID:16448655

  5. Simultaneous determination of neonicotinoid insecticides in human serum and urine using diatomaceous earth-assisted extraction and liquid chromatography-tandem mass spectrometry.

    PubMed

    Yamamuro, Tadashi; Ohta, Hikoto; Aoyama, Mika; Watanabe, Daisuke

    2014-10-15

    A rapid and sensitive analytical method was developed for simultaneous determination of eight neonicotinoid insecticides (acetamiprid, clothianidin, dinotefuran, flonicamid, imidacloprid, nitenpyram, thiacloprid and thiamethoxam) and three specific metabolites of acetamiprid (N-desmethylacetamiprid, 5-(N-acetyl-N-methylaminomethyl)-2-chloropyridine and 5-(N-acetylaminomethyl)-2-chloropyridine) in human serum and urine. A diatomaceous earth-assisted extraction using Extrelut NT3 column with chloroform/2-propanol (3:1, v/v) as eluent was selected for the single step cleanup procedure for all the target compounds. Qualitative and quantitative analyses were conducted by liquid chromatography-tandem mass spectrometry with multiple reaction monitoring mode. The limits of detection and the limits of quantification of eleven compounds were in the ranges of 0.1-0.2ng/mL and 0.5-10ng/mL for serum, 0.1-1ng/mL and 1-10ng/mL for urine, respectively. The extraction recoveries were between 80.9% and 101.8% for serum samples, 91.9% and 106% for urine samples. The intra-day RSDs and the inter-day RSDs were less than 11.5% and 13.2% for serum, less than 8.3% and 8.8% for urine. The proposed procedure will be suitable for forensic investigations of human poisoning cases with neonicotinoid insecticides. This is the first report of simultaneous determination of eight neonicotinoids in serum and urine samples.

  6. Micro-solid phase extraction with liquid chromatography-tandem mass spectrometry for the determination of aflatoxins in coffee and malt beverage.

    PubMed

    Khayoon, Wejdan Shakir; Saad, Bahruddin; Salleh, Baharuddin; Manaf, Normaliza Hj Abdul; Latiff, Aishah A

    2014-03-15

    A single step extraction-cleanup procedure using porous membrane-protected micro-solid phase extraction (μ-SPE) in conjunction with liquid chromatography-tandem mass spectrometry for the extraction and determination of aflatoxins (AFs) B1, B2, G1 and G2 from food was successfully developed. After the extraction, AFs were desorbed from the μ-SPE device by ultrasonication using acetonitrile. The optimum extraction conditions were: sorbent material, C8; sorbent mass, 20mg; extraction time, 90 min; stirring speed, 1,000 rpm; sample volume, 10 mL; desorption solvent, acetonitrile; solvent volume, 350 μL and ultrasonication period, 25 min without salt addition. Under the optimum conditions, enrichment factor of 11, 9, 9 and 10 for AFG2, AFG1, AFB2 and AFB1, respectively were achieved. Good linearity and correlation coefficient was obtained over the concentration range of 0.4-50 ng g(-1) (r(2) 0.9988-0.9999). Good recoveries for AFs ranging from 86.0-109% were obtained. The method was applied to 40 samples involving malt beverage (19) and canned coffee (21). No AFs were detected in the selected samples.

  7. Conventional and micellar liquid chromatography method development for danazol and validation in capsules.

    PubMed

    Gonzalo-Lumbreras, R; Izquierdo-Hornillos, R

    2003-07-14

    Two isocratic liquid chromatographic methods (conventional and micellar) for the determination of danazol (DZ) in capsules using canrenone (CAN) as internal standard have been developed and validated. In conventional liquid chromatography a mobile phase 35% water:acetonitrile 65%, v:v, a flow-rate 1 ml min(-1) and a C18 Hypersil ODS (250 x 4.6 mm, 5 microm) column (25 degrees C) were used. In micellar liquid chromatography (MLC) the conditions were: mobile phase 40 mM sodium dodecyl sulfate:2% pentanol, flow-rate 0.5 ml min(-1) and C18 Hypersil ODS (150 x 3.0 mm, 5 microm) column (60 degrees C). For both methods. UV absorbance detection at 280 nm was used and a separation up to base line was achieved. Prior to HPLC analysis a simple sample preparation was required. The recoveries found in the accuracy test were 99 +/- 10 and 101 +/- 8%, in conventional liquid chromatography (CLC) and MLC, respectively. Repeatability and intermediate precision expressed as R.S.D. were lower than 5% for both methods. Detection limits obtained were 2.4 and 3.0 ng g(-1) in CLC and CLM, respectively. PMID:14565547

  8. [An analysis of maicaodi by high performance liquid chromatography].

    PubMed

    Yang, H; Chen, R; Jiang, M

    1997-05-01

    Maicaodi has recently been developed and produced by the pesticide plant of Nanjing Agricultural University. The quantitative analysis of the effective components--tribenuron methyl and R (-)napropamide in wettable powder of Maicaode, by a high performance liquid chromatographic method was carried out with a Lichrosorb Si-60 20cm x 0.46cm i.d. column, mobile phase of petroleum ether/isopropanol/methanol/acetonitrile/chloroform mixture solvent (80:5:5:5:5) and internal standard of diisooctyl phthalate. The sample was detected by ultraviolet absorption at 254 nm. The retention times of tribenuron methyl and R (-)napropamide were 10-11min and 6-7min respectively. The coefficient of variation of this analysis was 0.34% with a recovery of 99.51%-100.32%. The coefficient of linear correlation was 0.9999. PMID:15739379

  9. Magnetic ionic liquid-based dispersive liquid-liquid microextraction for the determination of triazine herbicides in vegetable oils by liquid chromatography.

    PubMed

    Wang, Yuanpeng; Sun, Ying; Xu, Bo; Li, Xinpei; Jin, Rui; Zhang, Hanqi; Song, Daqian

    2014-12-19

    Magnetic ionic liquid-based dispersive liquid-liquid microextraction (MIL-based DLLME) was developed for extracting triazine herbicides from vegetable oils. The MIL, 1-hexyl-3-methylimidazolium tetrachloroferrate ([C6mim] [FeCl4]), was used as the microextraction solvent. The magnetic separation time was shortened by simply mixing carbonyl iron powder with the MIL in the sample after DLLME. The effects of several important experimental parameters, including the amount of MIL, the time of ultrasonic extraction, the type and the volume of cleanup solvent were investigated. The MIL-based DLLME coupled with liquid chromatography gave the limits of detection of 1.31-1.49ngmL(-1) and limits of quantification of 4.33-4.91 ng mL(-1) for triazine herbicides. When the present method was applied to the analysis of vegetable oil samples, the obtained recoveries were in the range of 81.8-114.2% and the relative standard deviations were lower than 7.7%. Compared with existing methods, the performances achieved by the present method were acceptable.

  10. Quantitation of antioxidants in water samples using ionic liquid dispersive liquid-liquid microextraction followed by high-performance liquid chromatography-ultraviolet detection.

    PubMed

    Sobhi, Hamid Reza; Kashtiaray, Amir; Farahani, Hadi; Farahani, Mohammad Reza

    2011-01-01

    A simple and efficient method, ionic liquid-based dispersive liquid-liquid microextraction combined with high-performance liquid chromatography-ultraviolet detection (HPLC-UV), has been applied for the extraction and determination of some antioxidants (Irganox 1010, Irganox 1076 and Irgafos 168) in water samples. The microextraction efficiency factors were investigated and optimized: 1-hexyl-3-methylimidazolium hexafluorophosphate [C(6)MIM][PF(6)] (0.06 g) as extracting solvent, methanol (0.5 mL) as disperser solvent without salt addition. Under the selected conditions, enrichment factors up to 48-fold, limits of detection (LODs) of 5.0-10.0 ng/mL and dynamic linear ranges of 25-1500 ng/mL were obtained. A reasonable repeatability (RSD≤11.8%, n=5) with satisfactory linearity (r(2)≥0.9954) of the results illustrated a good performance of the presented method. The accuracy of the method was tested by the relative recovery experiments on spiked samples, with results ranging from 85 to 118%. Finally, the method was successfully applied for determination of the analytes in several real water samples.

  11. Application of liquid-liquid-liquid microextraction and high-performance liquid-chromatography for the determination of sulfonamides in water.

    PubMed

    Lin, Che-Yi; Huang, Shang-Da

    2008-03-31

    This work presents a novel liquid-liquid-liquid microextraction (LLLME) technique for the extraction of sulfonamides from aqueous systems; it combines with high-performance liquid-chromatography-ultraviolet absorbance detection (HPLC/UV). In this experiment the sulfonamides were successively extracted from a donor phase (i.e., a water sample) into several microliters of an organic phase and then from the organic phase into an acceptor phase (i.e., an aqueous extract) by LLLME. The following separation and quantitative analyses were performed using HPLC/UV with 265 nm detection. Extraction condition such as solvent identity, agitation, extraction time, acceptor phase NaOH concentration, donor phase pH, and salt addition were optimized. Relative standard deviation (RSD, 2.6-5.3%), coefficient of estimation (R2, 0.9972-0.9999), and method detection limit (MDL, 0.11-0.77 ng mL(-1)) were achieved under the selected conditions. The proposed method was successfully applied to the analyses of three practical water samples and the relative recoveries of sulfonamides from the spiked water samples were in the range of 86.2-108.7%. The proposed method also confirms microextraction to be robust to monitoring trace levels of sulfacetamide, sulfadiazine, sulfathiazole, sulfamerazine, sulfadimidine, sulfamonomethoxine, sulfamethoxazole, and sulfaquinoxaline in aqueous samples. PMID:18331856

  12. Ionic-liquid-based dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography for the forensic determination of methamphetamine in human urine.

    PubMed

    Wang, Ruifeng; Qi, Xiujuan; Zhao, Lei; Liu, Shimin; Gao, Shuang; Ma, Xiangyuan; Deng, Youquan

    2016-07-01

    Determination of methamphetamine in forensic laboratories is a major issue due to its health and social harm. In this work, a simple, sensitive, and environmentally friendly method based on ionic liquid dispersive liquid-liquid microextraction combined with high-performance liquid chromatography was established for the analysis of methamphetamine in human urine. 1-Octyl-3-methylimidazolium hexafluorophosphate with the help of disperser solvent methanol was selected as the microextraction solvent in this process. Various parameters affecting the extraction efficiency of methamphetamine were investigated systemically, including extraction solvent and its volume, disperser solvent and its volume, sample pH, extraction temperature, and centrifugal time. Under the optimized conditions, a good linearity was obtained in the concentration range of 10-1000 ng/mL with determination coefficient >0.99. The limit of detection calculated at a signal-to-noise ratio of 3 was 1.7 ng/mL and the relative standard deviations for six replicate experiments at three different concentration levels of 100, 500, and 1000 ng/mL were 6.4, 4.5, and 4.7%, respectively. Meanwhile, up to 220-fold enrichment factor of methamphetamine and acceptable extraction recovery (>80.0%) could be achieved. Furthermore, this method has been successfully employed for the sensitive detection of a urine sample from a suspected drug abuser.

  13. Direct Measurement of Free Estradiol in Human Serum and Plasma by Equilibrium Dialysis-Liquid Chromatography-Tandem Mass Spectrometry.

    PubMed

    Ray, Julie A; Kushnir, Mark M; Rockwood, Alan L; Meikle, A Wayne

    2016-01-01

    We describe a direct method of measurement of free estradiol using equilibrium dialysis followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Serum aliquots and internal standards are extracted by liquid-liquid extraction using methyl-tert-butyl ether (MTBE) followed by derivatization with dansyl chloride. An API 5500 mass spectrometer operated in positive electrospray mode is used for detection.

  14. High-performance liquid chromatography coupled to ion spray mass spectrometry for the determination of colchicine at ppb levels in human biofluids.

    PubMed

    Tracqui, A; Kintz, P; Ludes, B; Rougé, C; Douibi, H; Mangin, P

    1996-01-26

    An original method based upon high-performance liquid chromatography coupled to ion spray mass spectrometry (HPLC-ISP-MS) has been developed for the identification and quantification of colchicine (COL) in human blood, plasma or urine. After single-step liquid-liquid extraction by dichloromethane at pH 8.0 using tofisopam (TOF) as an internal standard, solutes are separated on a 5-microns C18 Microbore (Alltech) column (250 x 1.0 mm, I.D.), using acetonitrile-2 mM NH4COOH, pH 3 buffer (75: 25, v/v) as the mobile phase (flow-rate 50 microliters/min). Detection is done by a Perkin-Elmer Sciex API-100 mass analyzer equipped with a ISP interface (nebulizing and curtain gas: N2, quality U; main settings: ISP, +4.0 kV; OR, +50 V; Q0, -10 V; Q1, -13 V; electron multiplier, +2.2 kV); MS data are collected as either total ion current (TIC, m/z 100-500 or 380-405), or selected ion monitoring (SIM) at m/z 400 and 383 for COL and TOF, respectively. COL mass spectrum shows a prominent molecular ion [M + H]+ at m/z 400. Increasing OR potential fails to provide a significant fragmentation. Retention times are 2.70 and 4.53 min for COL and TOF, respectively. The quantification method shows a good linearity (r = 0.998) over a concentration range from 5 to 200 ng/ml. The lower limit of detection in SIM mode is 0.6 ng/ml COL, making the method convenient for both clinical and forensic purposes.

  15. Liquid chromatography/tandem mass spectrometry method for simultaneous determination of cocaine and its metabolite (-)ecgonine methyl ester in human acidified stabilized plasma samples.

    PubMed

    Liu, Yongzhen; Zheng, Bo; Strafford, Stephanie; Orugunty, Ravi; Sullivan, Michael; Gus, Jeffrey; Heidbreder, Christian; Fudala, Paul J; Nasser, Azmi

    2014-06-15

    Two simple, sensitive and rapid liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) methods (low range and high range) were developed and validated for the quantification of cocaine and its metabolite (-)ecgonine methyl ester (EME) in human acidified stabilized plasma samples. In the low range assay, cocaine and the internal standard, cocaine-D3, were extracted using a single step liquid-liquid extraction from human acidified stabilized plasma. For the high range assay, human acidified stabilized plasma containing cocaine, EME, and the internal standards, cocaine-D3 and EME-D3, was mixed with acetonitrile, and the protein precipitate was separated by centrifugation. Both cocaine and EME extracted from both assays were separated on a HILIC column and detected in positive ion mode using multiple reaction monitoring (MRM). Both methods were validated and the specificity, linearity, lower limit of quantitation (LLOQ), precision, accuracy, recoveries and stability were determined. The linear range for the low range assay was 0.01-5ng/mL for cocaine; in the high range assay values were 5-1000ng/mL for cocaine and 1-200ng/mL for EME. The correlation coefficient (R(2)) values for both assays were 0.993 or greater. The precision and accuracy for intra-day and inter-day were better than 13.0%. The recovery was above 85% and matrix effects were low with the matrix factor ranging from 0.817 to 1.10 for both analytes in both assays. The validated methods were successfully used to quantify the plasma concentrations of cocaine and EME in clinical pharmacokinetic and pharmacodynamic studies.

  16. Rhamnolipid biosurfactant analysis using online turbulent flow chromatography-liquid chromatography-tandem mass spectrometry.

    PubMed

    Behrens, Beate; Helmer, Patrick O; Tiso, Till; Blank, Lars M; Hayen, Heiko

    2016-09-23

    Rhamnolipids are biosurfactants produced by a variety of bacterial species that present a promising alternative to surfactants from petrochemical or oleochemical origin. The success of the fermentation is evaluated by subsequent qualitative and quantitative analysis. However, the sample preparation for high numbers of samples is often laborious and inefficient. In this study an online sample preparation is developed for the qualitative and quantitative analysis of rhamnolipids by LC-MS/MS. Online sample preparation is carried out on a TurboFlow Cyclone MAX column using turbulent flow chromatography. Sample preparation prior the analysis is minimized to a dilution and syringe filtration step leading to an instrumental analysis time of 33min. The limit of detection and the limit of quantification were 0.4ng and 0.6ng on column, respectively. Recovery of the main mono- and di-rhamnolipids from a fermentation sample was 102-104%. Additionally, the rhamnolipid biosynthetic precursors 3-hydroxy(alkanoyloxy)alkanoic acids (HAAs) are covered, albeit extraction is not quantitative (85-90%). The analysis of rhamnolipids from four different microbial species was in good agreement with previous reports. The presented method allows rapid and comprehensive analysis of rhamnolipids with minimal sample preparation directly from the fermentation broth. The application of complementary data-dependent MS/MS acquisition enables non-target screening of rhamnolipids.

  17. Rhamnolipid biosurfactant analysis using online turbulent flow chromatography-liquid chromatography-tandem mass spectrometry.

    PubMed

    Behrens, Beate; Helmer, Patrick O; Tiso, Till; Blank, Lars M; Hayen, Heiko

    2016-09-23

    Rhamnolipids are biosurfactants produced by a variety of bacterial species that present a promising alternative to surfactants from petrochemical or oleochemical origin. The success of the fermentation is evaluated by subsequent qualitative and quantitative analysis. However, the sample preparation for high numbers of samples is often laborious and inefficient. In this study an online sample preparation is developed for the qualitative and quantitative analysis of rhamnolipids by LC-MS/MS. Online sample preparation is carried out on a TurboFlow Cyclone MAX column using turbulent flow chromatography. Sample preparation prior the analysis is minimized to a dilution and syringe filtration step leading to an instrumental analysis time of 33min. The limit of detection and the limit of quantification were 0.4ng and 0.6ng on column, respectively. Recovery of the main mono- and di-rhamnolipids from a fermentation sample was 102-104%. Additionally, the rhamnolipid biosynthetic precursors 3-hydroxy(alkanoyloxy)alkanoic acids (HAAs) are covered, albeit extraction is not quantitative (85-90%). The analysis of rhamnolipids from four different microbial species was in good agreement with previous reports. The presented method allows rapid and comprehensive analysis of rhamnolipids with minimal sample preparation directly from the fermentation broth. The application of complementary data-dependent MS/MS acquisition enables non-target screening of rhamnolipids. PMID:27567141

  18. Determination of chloroacetanilide herbicide metabolites in water using high-performance liquid chromatography-diode array detection and high-performance liquid chromatography/mass spectrometry

    USGS Publications Warehouse

    Hostetler, K.A.; Thurman, E.M.

    2000-01-01

    Analytical methods using high-performance liquid chromatography-diode array detection (HPLC-DAD) and high-performance liquid chromatography/mass spectrometry (HPLC/MS) were developed for the analysis of the following chloroacetanilide herbicide metabolites in water: alachlor ethanesulfonic acid (ESA); alachlor oxanilic acid; acetochlor ESA; acetochlor oxanilic acid; metolachlor ESA; and metolachlor oxanilic acid. Good precision and accuracy were demonstrated for both the HPLC-DAD and HPLC/MS methods in reagent water, surface water, and ground water. The average HPLC-DAD recoveries of the chloroacetanilide herbicide metabolites from water samples spiked at 0.25, 0.5 and 2.0 ??g/l ranged from 84 to 112%, with relative standard deviations of 18% or less. The average HPLC/MS recoveries of the metabolites from water samples spiked at 0.05, 0.2 and 2.0 ??g/l ranged from 81 to 118%, with relative standard deviations of 20% or less. The limit of quantitation (LOQ) for all metabolites using the HPLC-DAD method was 0.20 ??g/l, whereas the LOQ using the HPLC/MS method was at 0.05 ??g/l. These metabolite-determination methods are valuable for acquiring information about water quality and the fate and transport of the parent chloroacetanilide herbicides in water. Copyright (C) 2000 Elsevier Science B.V.

  19. [Simultaneous determination of eight furocoumarines in cosmetics by high performance liquid chromatography and verification by liquid chromatography-tandem mass spectrometry].

    PubMed

    Ma, Huijuan; Ma, Qiang; Li, Wentao; Meng, Xianshuang; Li, Jingrui; Bai, Hua; Jiao, Yang; Zhang, Xiaoli

    2013-05-01

    A method using high performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the simultaneous determination of eight furocoumarines (8-hydroxypsoralen, psoralen, isopsoralen, 8-methoxypsoralen, 5-methoxypsoralen, trioxsalen, imperatorin and isoimperatorin) in cosmetics. The cosmetic samples, including cream, lotion, shampoo, powder and lipstick, were supersonically extracted with appropriate solvents. The extract was centrifuged, and the supernatant was filtered through a membrane, and then separated on an Agilent Zorbax SB-Phenyl chromatographic column (250 mm x 4.6 mm, 5 microm) by gradient elution at a flow rate of 1.0 mL/min with methanol-acetonitrile-water as mobile phases. The column temperature was set at 30 degrees C. The wavelength of detection was 250 nm. The limits of quantification (LOQs) were 0.25 mg/kg for 8-hydroxypsoralen and 0.5 mg/kg for psoralen, isopsoralen, 8-methoxypsoralen, 5-methoxypsoralen, trioxsalen, imperatorin and isoimperatorin. The recoveries at three spiked levels were in the range of 85.0% - 105.8% with the relative standard deviations (RSDs) of 0.41% - 7.90%. The intra-day precision (n=6) was less than 1%, and the inter-day precision (n = 6) was less than 2% for the peak areas of the eight furocoumarines in a mixed standard solution. The method is accurate, simple, rapid and suitable for the determination of the eight furocoumarines in various cosmetic samples.

  20. [Simultaneous determination of eight furocoumarines in cosmetics by high performance liquid chromatography and verification by liquid chromatography-tandem mass spectrometry].

    PubMed

    Ma, Huijuan; Ma, Qiang; Li, Wentao; Meng, Xianshuang; Li, Jingrui; Bai, Hua; Jiao, Yang; Zhang, Xiaoli

    2013-05-01

    A method using high performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the simultaneous determination of eight furocoumarines (8-hydroxypsoralen, psoralen, isopsoralen, 8-methoxypsoralen, 5-methoxypsoralen, trioxsalen, imperatorin and isoimperatorin) in cosmetics. The cosmetic samples, including cream, lotion, shampoo, powder and lipstick, were supersonically extracted with appropriate solvents. The extract was centrifuged, and the supernatant was filtered through a membrane, and then separated on an Agilent Zorbax SB-Phenyl chromatographic column (250 mm x 4.6 mm, 5 microm) by gradient elution at a flow rate of 1.0 mL/min with methanol-acetonitrile-water as mobile phases. The column temperature was set at 30 degrees C. The wavelength of detection was 250 nm. The limits of quantification (LOQs) were 0.25 mg/kg for 8-hydroxypsoralen and 0.5 mg/kg for psoralen, isopsoralen, 8-methoxypsoralen, 5-methoxypsoralen, trioxsalen, imperatorin and isoimperatorin. The recoveries at three spiked levels were in the range of 85.0% - 105.8% with the relative standard deviations (RSDs) of 0.41% - 7.90%. The intra-day precision (n=6) was less than 1%, and the inter-day precision (n = 6) was less than 2% for the peak areas of the eight furocoumarines in a mixed standard solution. The method is accurate, simple, rapid and suitable for the determination of the eight furocoumarines in various cosmetic samples. PMID:24010339

  1. Evaluation of the Kinetic Performance Differences between Hydrophilic-Interaction Liquid Chromatography and Reversed-Phase Liquid Chromatography under Conditions of Identical Packing Structure.

    PubMed

    Song, Huiying; Desmet, Gert; Cabooter, Deirdre

    2015-12-15

    A protocol using trifluoroacetic acid at a temperature of 60 °C is developed for the adequate removal of the stationary phase of reversed-phase liquid chromatography (RPLC) columns. This procedure allows for studying the same column first under RPLC and subsequently under hydrophilic-interaction liquid chromatography (HILIC) conditions to isolate intrinsic differences between mass transfer properties in HILIC and RPLC from differences in packing quality. The established procedure allows for a complete removal of the stationary phase (confirmed by retention studies and thermogravimetry analyses) while leaving the structure of the packing unaffected (witnessed by an unchanged external porosity and pressure drop). Accurate plate height analysis comparing compounds at the same zone retention factor indicates a significant difference in reduced c-term (typically 40-80% larger under HILIC conditions), despite the columns otherwise being identical. Correcting for the known contributions of longitudinal diffusion (b-term) and mass transfer (cm- and cs-term) to focus on band broadening originating from eddy dispersion, similar strong differences are observed (differences of some h = 0.3 up to 1.2). These findings show that the interior structure and retention mechanism of the particles have a very strong effect on the observed eddy dispersion, a factor typically ascribed to phenomena occurring outside the particles. This also implies that comparing the quality of packings of different particle types is virtually impossible without the availability of a sound model to correct for the intraparticle effect on the observed eddy dispersion.

  2. Determination of nitrofurans in animal feeds by liquid chromatography-UV photodiode array detection and liquid chromatography-ionspray tandem mass spectrometry.

    PubMed

    Barbosa, Jorge; Moura, Sara; Barbosa, Rita; Ramos, Fernando; da Silveira, Maria Irene Noronha

    2007-03-14

    Within the EU, the use of nitrofurans is prohibited in food production animals. For this reason detection of these compounds in feedingstuffs, at whatever limit, constitutes an offence under EU legislation. This detection generally involves the use of analytical methods with limits of quantification lowers than 1 mg kg(-1). These procedures are unsuitable for the detection and confirmation of trace amounts of nitrofurans in feedingstuffs due to contamination. It is well known that very low concentrations of these compounds can be the source of residues of nitrofuran metabolites in meat and other edible products obtained from animals consuming the contaminated feed. The present multi-compound method was capable of measuring very low concentrations of nitrofurantoin (NFT), nitrofurazone (NFZ), furazolidone (FZD) and furaltadone (FTD) in animal feed using nifuroxazide (NXZ) as internal standard. Following ethyl acetate extraction at mild alkaline conditions and purification on NH2 column, the nitrofurans are determined using liquid chromatography with photodiode-array detection (LC-DAD). It was observed a CCalpha ranged from 50 to 100 microg kg(-1). The liquid chromatography-tandem mass spectrometric (LC-MS/MS) procedure was used to confirm the identity of the suspected presence of any of the nitrofuran compounds. PMID:17386735

  3. Simultaneous determination of fluoroquinolones in foods of animal origin by a high performance liquid chromatography and a liquid chromatography tandem mass spectrometry with accelerated solvent extraction.

    PubMed

    Yu, Huan; Tao, Yanfei; Chen, Dongmei; Pan, Yuanhu; Liu, Zhenli; Wang, Yulian; Huang, Lingli; Dai, Menghong; Peng, Dapeng; Wang, Xu; Yuan, Zonghui

    2012-02-15

    A confirmatory and quantitative method based on a high performance liquid chromatography UV detector (HPLC-UV) and a liquid chromatography tandem mass spectrometry (LC-MS/MS) with an extraction procedure of accelerated solvent extraction (ASE) has been developed for simultaneous determination of 15 kinds of fluoroquinolones in various animal origin food samples. The sample preparation procedures consist of an extraction step with acetonitrile and a cleaning-up step with Oasis HLB cartridge. Parameters for extraction pressure and temperature, cycle of ASE, clean-up, and analysis procedure have been optimized systematically. The recoveries of FQNs spiked in the tissues as the muscle, liver, kidney of swine, bovine, chicken and fish at a concentration range of 10-800μg/kg were found between 70.6% and 111.1% with relative standard deviations (RSD) less than 15% in HPLC. The LOD and LOQ of the HPLC for the 15 FQNs were 3μg/kg and 10μg/kg, respectively, and those of the LC-MS/MS were 0.3 and 1μg/kg, respectively. These rapid and reliable methods can be used to efficiently separate, characterize and quantify the residues of 15 FQNs (Marbofloxacin, Enoxacin, Fleroxacin, Ofloxacin, Pefloxacin, Lomefloxacin, Danofloxacin, Enrofloxacin, Orbifloxacin, Cinoxacin, Gatifloxacin, Sarafloxacin, Difloxacin, Nalidixic Acid, Flumequine) in food of animal origin.

  4. Improved characterization of tomato polyphenols using liquid chromatography/electrospray ionization linear ion trap quadrupole Orbitrap mass spectrometry and liquid chromatography/electrospray ionization tandem mass spectrometry.

    PubMed

    Vallverdú-Queralt, Anna; Jáuregui, Olga; Medina-Remón, Alexander; Andrés-Lacueva, Cristina; Lamuela-Raventós, Rosa M

    2010-10-30

    Tomato (Lycopersicon esculentum Mill.) is the second most important fruit crop worldwide. Tomatoes are a key component in the Mediterranean diet, which is strongly associated with a reduced risk of chronic degenerative diseases. In this work, we use a combination of mass spectrometry (MS) techniques with negative ion detection, liquid chromatography/electrospray ionization linear ion trap quadrupole-Orbitrap-mass spectrometry (LC/ESI-LTQ-Orbitrap-MS) and liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) on a triple quadrupole, for the identification of the constituents of tomato samples. First, we tested for the presence of polyphenolic compounds through generic MS/MS experiments such as neutral loss and precursor ion scans on the triple quadrupole system. Confirmation of the compounds previously identified was accomplished by injection into the high-resolution system (LTQ-Orbitrap) using accurate mass measurements in MS, MS(2) and MS(3) modes. In this way, 38 compounds were identified in tomato samples with very good mass accuracy (<2 mDa), three of them, as far as we know, not previously reported in tomato samples.

  5. Liquid chromatography-mass spectrometric and liquid chromatography-tandem mass spectrometric determination of hallucinogenic indoles psilocin and psilocybin in "magic mushroom" samples.

    PubMed

    Kamata, Tooru; Nishikawa, Mayumi; Katagi, Munehiro; Tsuchihashi, Hitoshi

    2005-03-01

    Accurate and sensitive analytical methods for psilocin (PC) and psilocybin (PB), tryptamine-type hallucinogens contained in "magic mushrooms," were investigated using liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS-MS). The chromatographic separation on an ODS column and mass spectral information gave complete discrimination between PC and PB without derivatization. The mass spectrometric detection had a high sensitivity, and the tandem mass spectrometric detection provided more specificity and accuracy, as well as high sensitivity. The detection limits ranged from 1 to 25 pg by LC-MS in the selected ion monitoring mode, and the intra- and inter-day coefficients of variation were estimated to be 4.21-5.93% by LC-MS-MS in the selected reaction monitoring mode. By applying the present LC-MS-MS technique to four real samples, the contents of PC and PB were found to vary over a wide range (0.60-1.4 and 0.18-3.8 mg/g dry wt. for PC and PB, respectively) between samples.

  6. Separation and characterization of bufadienolides in toad skin using two-dimensional normal-phase liquid chromatography×reversed-phase liquid chromatography coupled with mass spectrometry.

    PubMed

    Zhang, Yun; Jin, Hongli; Li, Xiaolong; Zhao, Jianqiang; Guo, Xiujie; Wang, Jixia; Guo, Zhimou; Zhang, Xiuli; Tao, Yanduo; Liu, Yanfang; Chen, Deliang; Liang, Xinmiao

    2016-07-15

    Bufadienolides possess various bioactivities especially antitumor. Due to the high structural diversity, the separation of bufadienolides often suffers from coelution problem on conventional RP columns. In this work, an off-line two-dimensional normal-phase liquid chromatography×reversed-phase liquid chromatography (2D-NPLC×RPLC) method was developed to separate and characterize bufadienolides in toad skin. Several RP and NP columns were evaluated with five reference bufadienlides. The XUnion C18 and XAmide columns exhibited superior chromatographic performances for bufadienlide separation, and were selected in RPLC and NPLC, respectively. RPLC was used in the second-dimension for the good compatibility with MS, while NPLC was adopted in the first-dimension. The orthogonality of the 2D-NPLC×RPLC system was investigated by the geometric approach using fifteen bufadienolide mixtures. The result was 49.6%, demonstrating reasonable orthogonality of this 2D-LC system. By combining the 2D-LC system with MS, 64 bufadienlides including 33 minor ones and 11 pairs of isomers in toad skin were identified. This off-line 2D-NPLC×RPLC allowed to solve the coelution problem of bufadienlides in one-dimension RPLC, and thus facilitated the identification significantly.

  7. Evaluation of the Kinetic Performance Differences between Hydrophilic-Interaction Liquid Chromatography and Reversed-Phase Liquid Chromatography under Conditions of Identical Packing Structure.

    PubMed

    Song, Huiying; Desmet, Gert; Cabooter, Deirdre

    2015-12-15

    A protocol using trifluoroacetic acid at a temperature of 60 °C is developed for the adequate removal of the stationary phase of reversed-phase liquid chromatography (RPLC) columns. This procedure allows for studying the same column first under RPLC and subsequently under hydrophilic-interaction liquid chromatography (HILIC) conditions to isolate intrinsic differences between mass transfer properties in HILIC and RPLC from differences in packing quality. The established procedure allows for a complete removal of the stationary phase (confirmed by retention studies and thermogravimetry analyses) while leaving the structure of the packing unaffected (witnessed by an unchanged external porosity and pressure drop). Accurate plate height analysis comparing compounds at the same zone retention factor indicates a significant difference in reduced c-term (typically 40-80% larger under HILIC conditions), despite the columns otherwise being identical. Correcting for the known contributions of longitudinal diffusion (b-term) and mass transfer (cm- and cs-term) to focus on band broadening originating from eddy dispersion, similar strong differences are observed (differences of some h = 0.3 up to 1.2). These findings show that the interior structure and retention mechanism of the particles have a very strong effect on the observed eddy dispersion, a factor typically ascribed to phenomena occurring outside the particles. This also implies that comparing the quality of packings of different particle types is virtually impossible without the availability of a sound model to correct for the intraparticle effect on the observed eddy dispersion. PMID:26595107

  8. Chemical fingerprint of Ganmaoling granule by double-wavelength ultra high performance liquid chromatography and ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry.

    PubMed

    Lou, Qiong; Ye, Xiaolan; Zhou, Yingyi; Li, Hua; Song, Fenyun

    2015-06-01

    A method incorporating double-wavelength ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry was developed for the investigation of the chemical fingerprint of Ganmaoling granule. The chromatographic separations were performed on an ACQUITY UPLC HSS C18 column (2.1 × 50 mm, 1.8 μm) at 30°C using gradient elution with water/formic acid (1%) and acetonitrile at a flow rate of 0.4 mL/min. A total of 11 chemical constituents of Ganmaoling granule were identified from their molecular weight, UV spectra, tandem mass spectrometry data, and retention behavior by comparing the results with those of the reference standards or literature. And 25 peaks were selected as the common peaks for fingerprint analysis to evaluate the similarities among 25 batches of Ganmaoling granule. The results of principal component analysis and orthogonal projection to latent structures discriminant analysis showed that the important chemical markers that could distinguish the different batches were revealed as 4,5-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4-O-caffeoylquinic acid. This is the first report of the ultra high performance liquid chromatography chemical fingerprint and component identification of Ganmaoling granule, which could lay a foundation for further studies of Ganmaoling granule.

  9. On-line comprehensive two-dimensional normal-phase liquid chromatography × reversed-phase liquid chromatography for preparative isolation of Peucedanum praeruptorum.

    PubMed

    Wang, Xin-Yuan; Li, Jia-Fu; Jian, Ya-Mei; Wu, Zhen; Fang, Mei-Juan; Qiu, Ying-Kun

    2015-03-27

    A new on-line comprehensive preparative two-dimensional normal-phase liquid chromatography × reversed-phase liquid chromatography (2D NPLC × RPLC) system was developed for the separation of complicated natural products. It was based on the use of a silica gel packed medium-pressure column as the first dimension and an ODS preparative HPLC column as the second dimension. The two dimensions were connected with normal-phase (NP) and reversed-phase (RP) enrichment units, involving a newly developed airflow assisted adsorption (AAA) technique. The instrument operation and the performance of this NPLC × RPLC separation method were illustrated by gram-scale isolation of ethanol extract from the roots of Peucedanum praeruptorum. In total, 19 compounds with high purity were obtained via automated multi-step preparative separation in a short period of time using this system, and their structures were comprehensively characterized by ESI-MS, (1)H NMR, and (13)C NMR. Including two new compounds, five isomers in two groups with identical HPLC and TLC retention values were also obtained and identified by 1D NMR and 2D NMR. This is the first report of an NPLC × RPLC system successfully applied in an on-line preparative process. This system not only solved the interfacing problem of mobile-phase immiscibility caused by NP and RP separation, it also exhibited apparent advantages in separation efficiency and sample treatment capacity compared with conventional methods.

  10. Hyphenated and comprehensive liquid chromatography × gas chromatography-mass spectrometry for the identification of Mycobacterium tuberculosis.

    PubMed

    Mourão, Marta P B; Denekamp, Ilse; Kuijper, Sjoukje; Kolk, Arend H J; Janssen, Hans-Gerd

    2016-03-25

    Tuberculosis is one of the world's most emerging public health problems, particularly in developing countries. Chromatography based methods have been used to tackle this epidemic by focusing on biomarker detection. Unfortunately, interferences from lipids in the sputum matrix, particularly cholesterol, adversely affect the identification and detection of the marker compounds. The present contribution describes the serial combination of normal phase liquid chromatography (NPLC) with thermally assisted hydrolysis and methylation followed by gas chromatography-mass spectrometry (THM-GC-MS) to overcome the difficulties of biomarker evaluation. The in-series combination consists of an LC analysis where fractions are collected and then transferred to the THM-GC-MS system. This was either done with comprehensive coupling, transferring all the fractions, or with hyphenated interfacing, i.e. off-line multi heart-cutting, transferring only selected fractions. Owing to the high sensitivity and selectivity of LC as a sample pre-treatment method, and to the high specificity of the MS as a detector, this analytical approach, NPLC × THM-GC-MS, is extremely sensitive. The results obtained indicate that this analytical set-up is able to detect down to 1 × 10(3) mycobacteria/mL of Mycobacterium tuberculosis strain 124, spiked in blank sputum samples. It is a powerful analytical tool and also has great potential for full automation. If further studies demonstrate its usefulness when applied blind in real sputum specimens, this technique could compete with the current smear microscopy in the early diagnosis of tuberculosis. PMID:26585206

  11. Online coupling of hydrophilic interaction/strong cation exchange/reversed-phase liquid chromatography with porous graphitic carbon liquid chromatography for simultaneous proteomics and N-glycomics analysis.

    PubMed

    Zhao, Yun; Law, Henry C H; Zhang, Zaijun; Lam, Herman C; Quan, Quan; Li, Guohui; Chu, Ivan K

    2015-10-01

    In this study we developed a fully automated three-dimensional (3D) liquid chromatography methodology-comprising hydrophilic interaction separation as the first dimension, strong cation exchange fractionation as the second dimension, and low-pH reversed-phase (RP) separation as the third dimension-in conjunction downstream with additional complementary porous graphitic carbon separation, to capture non-retained hydrophilic analytes, for both shotgun proteomics and N-glycomics analyses. The performance of the 3D system alone was benchmarked through the analysis of the total lysate of Saccharomyces cerevisiae, leading to improved hydrophilic peptide coverage, from which we identified 19% and 24% more proteins and peptides, respectively, relative to those identified from a two-dimensional hydrophilic interaction liquid chromatography and low-pH RP chromatography (HILIC-RP) system over the same mass spectrometric acquisition time; consequently, the 3D platform also provided enhanced proteome and protein coverage. When we applied the integrated technology to analyses of the total lysate of primary cerebellar granule neurons, we characterized a total of 2201 proteins and 16,937 unique peptides for this primary cell line, providing one of its most comprehensive datasets. Our new integrated technology also exhibited excellent performance in the first N-glycomics analysis of cynomolgus monkey plasma; we successfully identified 122 proposed N-glycans and 135 N-glycosylation sites from 122 N-glycoproteins, and confirmed the presence of 38 N-glycolylneuraminic acid-containing N-glycans, a rare occurrence in human plasma, through tandem mass spectrometry for the first time.

  12. Quantitative liquid chromatography/tandem mass spectrometry determination of chloramphenicol residues in food using sub-2 microm particulate high-performance liquid chromatography columns for sensitivity and speed.

    PubMed

    Kaufmann, Anton; Butcher, Patrick

    2005-01-01

    The use of chloramphenicol (CAP)--a highly effective broad-spectrum antibiotic used in animal husbandry--is banned in many countries. Therefore, a very low minimum required performance limit (MRPL) of 0.3 microg/kg CAP in meat for human consumption has been defined. Analytical methods capable of quantifying and confirming such low residue levels require sophisticated instrumentation. Preferably sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) or gas chromatography/mass spectrometry (GC/MS) methods have been used. This paper suggests the use of sub-2 microm particulate high-performance liquid chromatography (HPLC) columns to gain additional sensitivity and improve resolution as well as speed. Depending on the operating conditions, higher chromatographic resolution and speed can be obtained at the price of a significantly increased operating pressure, requiring dedicated LC equipment. A 3-4-fold overall improvement of the signal-to-noise ratio for CAP was obtained compared to more classical 5 microm particulate HPLC columns. The proposed analytical methodology includes an enzymatic digestion, which liberates glucuronide-bound CAP from kidney tissue. The extracts obtained after an Extrelut clean-up are sufficiently pure to permit routine injection of biological samples into the sub-2 microm particulate HPLC column, without observing rapid deterioration of peak shape or column clogging problems. The time for one chromatographic run was 4.2 min. The described method was validated for two particularly difficult matrices (kidney and honey). Decision limits (CC alpha) were 0.007 microg/kg (honey) und 0.011 microg/kg (kidney), which are significantly below the current MRPL. PMID:16299695

  13. Development, validation and determination of multiclass pesticide residues in cocoa beans using gas chromatography and liquid chromatography tandem mass spectrometry.

    PubMed

    Zainudin, Badrul Hisyam; Salleh, Salsazali; Mohamed, Rahmat; Yap, Ken Choy; Muhamad, Halimah

    2015-04-01

    An efficient and rapid method for the analysis of pesticide residues in cocoa beans using gas and liquid chromatography-tandem mass spectrometry was developed, validated and applied to imported and domestic cocoa beans samples collected over 2 years from smallholders and Malaysian ports. The method was based on solvent extraction method and covers 26 pesticides (insecticides, fungicides, and herbicides) of different chemical classes. The recoveries for all pesticides at 10 and 50 μg/kg were in the range of 70-120% with relative standard deviations of less than 20%. Good selectivity and sensitivity were obtained with method limit of quantification of 10 μg/kg. The expanded uncertainty measurements were in the range of 4-25%. Finally, the proposed method was successfully applied for the routine analysis of pesticide residues in cocoa beans via a monitoring study where 10% of them was found positive for chlorpyrifos, ametryn and metalaxyl. PMID:25442595

  14. [Trigonelline in coffee. I. Comparison of thin layer chromatography with high-performance liquid chromatography. Simultaneous determination of caffeine].

    PubMed

    Stennert, A; Maier, H G

    1993-05-01

    Determination of trigonelline in several green and roasted coffees by thin-layer (TLC) and high-performance liquid chromatography (HPLC) were compared. Using TLC, two methods of detection were also compared. In addition, caffeine could be determined using HPLC. The determinations were each made with several variations of extraction and clean-up. Recoveries, variation coefficients and detection limits are listed. The most suitable method was HPLC in connection with hot water extraction. Only water is required as the mobile phase. The values for precision are in most cases better than (trigonelline) or similar to (caffeine) those reported in the literature. The determination of trigonelline by TLC, followed by extraction and photometry, is possible with a similar precision, but the simultaneous determination of caffeine was not tested. PMID:8511975

  15. Development, validation and determination of multiclass pesticide residues in cocoa beans using gas chromatography and liquid chromatography tandem mass spectrometry.

    PubMed

    Zainudin, Badrul Hisyam; Salleh, Salsazali; Mohamed, Rahmat; Yap, Ken Choy; Muhamad, Halimah

    2015-04-01

    An efficient and rapid method for the analysis of pesticide residues in cocoa beans using gas and liquid chromatography-tandem mass spectrometry was developed, validated and applied to imported and domestic cocoa beans samples collected over 2 years from smallholders and Malaysian ports. The method was based on solvent extraction method and covers 26 pesticides (insecticides, fungicides, and herbicides) of different chemical classes. The recoveries for all pesticides at 10 and 50 μg/kg were in the range of 70-120% with relative standard deviations of less than 20%. Good selectivity and sensitivity were obtained with method limit of quantification of 10 μg/kg. The expanded uncertainty measurements were in the range of 4-25%. Finally, the proposed method was successfully applied for the routine analysis of pesticide residues in cocoa beans via a monitoring study where 10% of them was found positive for chlorpyrifos, ametryn and metalaxyl.

  16. Simultaneous achiral-chiral analysis of pharmaceutical compounds using two-dimensional reversed phase liquid chromatography-supercritical fluid chromatography.

    PubMed

    Venkatramani, C J; Al-Sayah, Mohammad; Li, Guannan; Goel, Meenakshi; Girotti, James; Zang, Lisa; Wigman, Larry; Yehl, Peter; Chetwyn, Nik

    2016-02-01

    A new interface was designed to enable the coupling of reversed phase liquid chromatography (RPLC) and supercritical fluid chromatography (SFC). This online two-dimensional chromatographic system utilizing RPLC in the first dimension and SFC in the second was developed to achieve simultaneous achiral and chiral analysis of pharmaceutical compounds. The interface consists of an eight-port, dual-position switching valve with small volume C-18 trapping columns. The peaks of interest eluting from the first RPLC dimension column were effectively focused as sharp concentration pulses on small volume C-18 trapping column/s and then injected onto the second dimension SFC column. The first dimension RPLC separation provides the achiral purity result, and the second dimension SFC separation provides the chiral purity result (enantiomeric excess). The results are quantitative enabling simultaneous achiral, chiral analysis of compounds. The interface design and proof of concept demonstration are presented. Additionally, comparative studies to conventional SFC and case studies of the applications of 2D LC-SFC in pharmaceutical analysis is presented.

  17. Phytochemical Profile of Erythrina variegata by Using High-Performance Liquid Chromatography and Gas Chromatography-Mass Spectroscopy Analyses.

    PubMed

    Muthukrishnan, Suriyavathana; Palanisamy, Subha; Subramanian, Senthilkumar; Selvaraj, Sumathi; Mari, Kavitha Rani; Kuppulingam, Ramalingam

    2016-08-01

    Natural products derived from plant sources have been utilized to treat patients with numerous diseases. The phytochemical constituents present in ethanolic leaf extract of Erythrina variegata (ELEV) were identified by using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectroscopy (GC-MS) analyses. Shade dried leaves were powdered and extracted with ethanol for analyses through HPLC to identify selected flavonoids and through GC-MS to identify other molecules. The HPLC analysis of ELEV showed the presence of gallic and caffeic acids as the major components at concentrations of 2.0 ppm and 0.1 ppm, respectively, as well as other components. GC-MS analysis revealed the presence of 3-eicosyne; 3,7,11,15-tetramethyl-2-hexadecen-1-ol; butanoic acid, 3-methyl-3,7-dimethyl-6-octenyl ester; phytol; 1,2-benzenedicarboxylic acid, diundecyl ester; 1-octanol, 2-butyl-; squalene; and 2H-pyran, 2-(7-heptadecynyloxy) tetrahydro-derivative. Because pharmacopuncture is a new evolving natural mode that uses herbal extracts for treating patients with various ailments with minimum pain and maximum effect, the results of this study are particularly important and show that ELEV possesses a wide range of phytochemical constituents, as indicated above, as effective active principle molecules that can be used individually or in combination to treat patients with various diseases. PMID:27555226

  18. Differentiation of coloured inks of inkjet printer cartridges by thin layer chromatography and high performance liquid chromatography.

    PubMed

    Poon, N L; Ho, S S H; Li, C K

    2005-01-01

    Document examiners are frequently asked to determine whether or not a colour printout has originated from a particular inkjet printer. The printer can rarely be identified unless some unique defects or irregularities of the printer are present on the printout. However, it is possible to decipher the make and/or model of the printer by comparing the ink-profile of the questioned printout with that of a seized inkjet printer cartridge or from one in a database. This paper presents an overview of a systematic approach to characterising and discriminating the inks of different inkjet printer cartridges using thin layer chromatography (TLC) and high pressure liquid chromatography (HPLC) with multi-wavelength ultra-violet/visible (UV/Vis) detection. Ink samples from 23 different inkjet printer cartridges (including originals and substitutes) of different brands and colour printouts, printed by known printers were examined with newly developed chromatographic methods. Subsequently, a database of the ink-profiles was generated. The methods provide a useful tool for discriminating coloured inks in inkjet printer cartridges of different brands.

  19. Liquid chromatography/microspray mass spectrometry for bacterial investigations.

    PubMed

    Krishnamurthy, T; Davis, M T; Stahl, D C; Lee, T D

    1999-01-01

    Cellular proteins (biomarkers) specific to any individual microorganism, determined by the direct mass spectral analysis of the corresponding intact cellular suspension, can be applied for the rapid and specific identification of the organisms present in unknown samples. The components of the bacterial suspensions, after a rapid separation over a C18 reversed-phase microcapillary column, were directly subjected to on-line electrospray ionization followed by analysis using an ion trap tandem mass spectrometer. This approach is equally effective for gram-positive as well as gram-negative bacteria but has a distinct advantage over our earlier reported method involving matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). During electrospray ionitation mass spectrometry (ESI-MS), liquid samples can be directly analyzed and there is the potential for developing tandem mass spectral methods for more specific identification of the individual organisms present in crude bacterial mixtures. The total analysis time leading to unambiguous bacterial identification in samples was less than 10 minutes and the results were quite reproducible. Miniaturization of the instrumentation along with total automation of this simple process could have immense impact on field operations. Routine, rapid, cost-effective field monitoring of environmental samples, agricultural products, samples from food processing, industrial sites and health institutions for suspected bacterial contamination could be a reality in the near future. Potential utility in biological, medical, bioprocessing, pharmaceutical, and other industrial research is also enormous.

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

    SciTech Connect

    Lee, Jung Hwa; Hyung, Seok-Won; Mun, Dong-Gi; Jung, Hee-Jung; Kim, Hokeun; Lee, Hangyeore; Kim, Su-Jin; Park, Kyong Soo; Moore, Ronald J.; Smith, Richard D.; Lee, Sang-Won

    2012-08-03

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

  1. Principles and Applications of Liquid Chromatography-Mass Spectrometry in Clinical Biochemistry

    PubMed Central

    Pitt, James J

    2009-01-01

    Liquid chromatography-mass spectrometry (LC-MS) is now a routine technique with the development of electrospray ionisation (ESI) providing a simple and robust interface. It can be applied to a wide range of biological molecules and the use of tandem MS and stable isotope internal standards allows highly sensitive and accurate assays to be developed although some method optimisation is required to minimise ion suppression effects. Fast scanning speeds allow a high degree of multiplexing and many compounds can be measured in a single analytical run. With the development of more affordable and reliable instruments, LC-MS is starting to play an important role in several areas of clinical biochemistry and compete with conventional liquid chromatography and other techniques such as immunoassay. PMID:19224008

  2. Alternative solvents for improving the greenness of normal phase liquid chromatography of lipid classes.

    PubMed

    Prache, Nolwenn; Abreu, Sonia; Sassiat, Patrick; Thiébaut, Didier; Chaminade, Pierre

    2016-09-16

    An evaluation of solvents alternative to n-heptane (d-limonene and hexamethyldisiloxane) and chloroform (cyclopentyl methyl ether, 2-methyltetrahydrofuran and isopentyl acetate) was developed for lipid classes separation of non-polar cholesteryl ester to highly polar phospholipids by high-performance liquid chromatography on bare silica stationary phase and evaporative light-scattering detection. Screening of alternative solvents was used to estimate their compatibility with liquid chromatography and evaporative light-scattering detection and to evaluate their chromatographic selectivity. This work shows that n-heptane can be advantageously replaced by hexamethyldisiloxane. An increase of non-polar lipids retention is observed with hexamethyldisiloxane as weak solvent. Chloroform, which is largely used for lipid analysis, might be replaced efficaciously by cyclopentyl methyl ether, 2-methyltetrahydrofuran or isopentyl acetate. Aside from offering a different selectivity, the gradients composed by one or both alternative solvents gave efficient and comparable or even better separations than those obtained with conventional solvents. PMID:27554026

  3. Analysis of antimycin A by reversed-phase liquid chromatography/nuclear magnetic-resonance spectrometry

    USGS Publications Warehouse

    Ha, Steven T.K.; Wilkins, Charles L.; Abidi, Sharon L.

    1989-01-01

    A mixture of closely related streptomyces fermentation products, antimycin A, Is separated, and the components are identified by using reversed-phase high-performance liquid chromatography with directly linked 400-MHz proton nuclear magnetic resonance detection. Analyses of mixtures of three amino acids, alanine, glycine, and valine, are used to determine optimal measurement conditions. Sensitivity increases of as much as a factor of 3 are achieved, at the expense of some loss in chromatographic resolution, by use of an 80-μL NMR cell, Instead of a smaller 14-μL cell. Analysis of the antimycin A mixture, using the optimal analytical high performance liquid chromatography/nuclear magnetic resonance conditions, reveals it to consist of at least 10 closely related components.

  4. Two-dimensional liquid chromatography system for online top-down mass spectrometry

    SciTech Connect

    Tian, Zhixin; Zhao, Rui; Tolic, Nikola; Moore, Ronald J.; Stenoien, David L.; Robinson, Errol W.; Smith, Richard D.; Pasa-Tolic, Ljiljana

    2010-10-01

    An online metal-free weak cation exchange-hydrophilic interaction liquid chromatography/reversed phase liquid chromatography (WCX-HILIC/RPLC) system has been developed for sensitive high-throughput top-down mass spectrometry. Analyzing posttranslational modifications (PTMs) of core histones, with focus on histone H4, tested the system. Using ~24 μg of core histones (H4, H2B, H2A and H3) purified from human fibroblasts, 41 H4 isoforms were identified, with the type and locations of PTMs unambiguously mapped for 20 of these variants. Compared to corresponding offline studies reported previously, online WCXHILIC/ RPLC platform offers significant improvement in sensitivity, with several orders of magnitude reduction in sample requirements and reduction in the overall analysis time. To the best of our knowledge, this study represents the first online two-dimensional (2D) LC-MS/MS characterization of core histone mixture at the intact protein level.

  5. Emerging liquid chromatography-mass spectrometry technologies improving dried blood spot analysis.

    PubMed

    Rao, Ramisetti Nageswara

    2014-08-01

    Dried blood spots (DBS), a micro blood sampling technique, has recently gained interest in drug discovery and development due to its inherent advantages over the conventional whole blood, plasma or serum sample collection. Since the regulatory authorities have agreed to the use of blood as an acceptable biological matrix for drug exposure measurements, its applications have been extended not only to therapeutic drug monitoring but also to toxicokinetic and pharmacokinetic studies. The pharmaceutical industry is keen to promote DBS as a prominent tool in bioanalytical applications due to the financial, ethical and organizational issues involved in clinical trials. This could be accomplished due to the latest advances in modern analytical technology, particularly liquid chromatography-mass spectrometry. The present review discusses some of the emerging liquid chromatography-mass spectrometry technologies in improving DBS analysis for its innovative applications in the development of new drugs.

  6. Gas-liquid chromatography in routine processing of blood cultures for detecting anaerobic bacteraemia.

    PubMed Central

    Reig, M; Molina, D; Loza, E; Ledesma, M A; Meseguer, M A

    1981-01-01

    Gas-liquid chromatography was performed on 233 positive blood cultures and findings were compared with culture results. Obligate anaerobic bacteria were recovered from 78 out of 79 blood cultures containing butyric or iso-valeric acids, or both; from 28 out of 69 blood cultures containing succinic acid; and from only one out of 41 blood cultures containing succinic but not butyric or iso-valeric acid. Good correlations (88%) were found for the recovery of anaerobic bacteria and the detection of butyric and/or iso-valeric acids. Detecting volatile fatty acids by gas-liquid chromatography performed on blood cultures at the first signs of growth can therefore provide an early and reliable indication of the presence of anaerobic bacteria. PMID:7014645

  7. Homochiral metal-organic framework used as a stationary phase for high-performance liquid chromatography.

    PubMed

    Kong, Jiao; Zhang, Mei; Duan, Ai-Hong; Zhang, Jun-Hui; Yang, Rui; Yuan, Li-Ming

    2015-02-01

    Metal-organic frameworks are promising porous materials. Chiral metal-organic frameworks have attracted considerable attention in controlling enantioselectivity. In this study, a homochiral metal-organic framework [Co(2) (D-cam)(2) (TMDPy)] (D-cam = D-camphorates, TMDPy = 4,4'-trimethylenedipyridine) with a non-interpenetrating primitive cubic net has been used as a chiral stationary phase in high-performance liquid chromatography. It has allowed the successful separation of six positional isomers and six chiral compounds. The good selectivity and baseline separation, or at least 60% valley separation, confirmed its excellent molecular recognition characteristics. The relative standard deviations for the retention time of run-to-run and column-to-column were less than 1.8 and 3.1%, respectively. These results demonstrate that [Co(2) (D-cam)(2) (TMDPy)] may represent a promising chiral stationary phase for use in high-performance liquid chromatography.

  8. Analyses of acute kidney injury biomarkers by ultra-high performance liquid chromatography with mass spectrometry.

    PubMed

    Al Za'abi, Mohammed; Ali, Badreldin H; ALOthman, Zeid A; Ali, Imran

    2016-01-01

    The newly developed acute kidney injury biomarkers are very important for the early and timely detection of kidney diseases. This review contains details of the analyses of several acute kidney injury biomarkers using ultra-high performance liquid chromatography-mass spectrometry in urine and plasma samples. In this review we attempt to discuss some aspects of the types of the biomarkers, patents, sample preparation, and the analyses. Besides, efforts were also made to discuss the possible uses of superficially porous (core-shell) columns in traditional and inexpensive high-performance liquid chromatography instruments. Additionally, the challenges and the future prospects are also highlighted. The present review will be useful for the academicians, scientists, and clinicians for the early detection of acute kidney injury biomarkers.

  9. Optimization of conditions for the single step IMAC purification of miraculin from Synsepalum dulcificum.

    PubMed

    He, Zuxing; Tan, Joo Shun; Lai, Oi Ming; Ariff, Arbakariya B

    2015-08-15

    In this study, the methods for extraction and purification of miraculin from Synsepalum dulcificum were investigated. For extraction, the effect of different extraction buffers (phosphate buffer saline, Tris-HCl and NaCl) on the extraction efficiency of total protein was evaluated. Immobilized metal ion affinity chromatography (IMAC) with nickel-NTA was used for the purification of the extracted protein, where the influence of binding buffer pH, crude extract pH and imidazole concentration in elution buffer upon the purification performance was explored. The total amount of protein extracted from miracle fruit was found to be 4 times higher using 0.5M NaCl as compared to Tris-HCl and phosphate buffer saline. On the other hand, the use of Tris-HCl as binding buffer gave higher purification performance than sodium phosphate and citrate-phosphate buffers in IMAC system. The optimum purification condition of miraculin using IMAC was achieved with crude extract at pH 7, Tris-HCl binding buffer at pH 7 and the use of 300 mM imidazole as elution buffer, which gave the overall yield of 80.3% and purity of 97.5%. IMAC with nickel-NTA was successfully used as a single step process for the purification of miraculin from crude extract of S. dulcificum. PMID:25794715

  10. Solvent systems for countercurrent chromatography: an aqueous two phase liquid system based on a room temperature ionic liquid.

    PubMed

    Ruiz-Angel, Maria Jose; Pino, Veronica; Carda-Broch, Samuel; Berthod, Alain

    2007-06-01

    A new aqueous two phase liquid system (ATPS) based on the ionic liquid 1-butyl-3-methyl imidazolium chloride (BMIM Cl), potassium dibasic phosphate (K(2)HPO(4)) and water was recently proposed in the literature. The full phase diagram of this ATPS was prepared and some tie lines were fully determined. It was compared to classical ATPSs based on polyethylene glycol with an average molecular mass of 1000 (PEG 1000) and 10,000 (PEG 10000) and K(2)HPO(4). Two countercurrent chromatography (CCC) columns, a hydrostatic Sanki and a J type hydrodynamic CCC columns were used to test the liquid phase retention of these ATPSs in all possible configurations. It was found that the BMIM Cl ATPS liquid phases were much easier to retain in the two CCC columns than the PEG 1000 ATPS phases. Using protein and alcohol solutes, it was established that the BMIM Cl ATPS has a polarity completely different from that of the PEG 1000 ATPS. For example, ovalbumin partitions equally between the two phases of the PEG 1000 ATPS (K(D)=1.4) when it is completely located in the BMIM Cl upper phase of the ionic liquid ATPS (K(D)=180). The discrimination factor of the ionic liquid system and its intrinsic hydrophobicity were respectively found three times higher and ten times lower than the respective values of the PEG 1000 ATPS. PMID:17166506

  11. [Determination of N-nitrosodimethylamine in beer by frozen zone melting liquid-liquid extraction/gas chromatography].

    PubMed

    Peng, Qiaorong; Tang, Tao; Yu, Shuxin; Sun, Yuanshe; Lei, Wu; Wang, Fengyun; Zhang, Weibing; Li, Tong

    2014-04-01

    A simple and effective sample enrichment method of frozen zone melting liquid-liquid extraction was optimized and validated for the analysis of trace N-nitrosodimethylamine (NDMA) in beer samples. The method was based on high pressure liquid-liquid extraction with a low temperature frozen step. The 90 mL beer was placed in a container with 10 mL dichloromethane. After agitation, the sample was kept in a freezer for 16 h at -19 degrees C. The organic extract was analyzed by gas chromatography with a flame ionization detector (GC-FID). The accuracy, precision, detection and quantification limits and linearity of the method were evaluated. The results showed that the calibration curve of NDMA was linear in the range of 5-200 mg/L with a good correlation coefficient (r2) of 0.999 6. The recoveries at the spiked levels of 5, 10 and 20 mg/L were 84.94%, 83.24%, 85.14% with the relative standard deviations (n = 7) of 3.06%, 3.19%, 2.63%, respectively. The ordinary extraction method of N-nitrosodimethylamine in beer includes the four steps of low-temperature distillation, liquid-liquid extraction, rotary evaporation and nitrogen blowing concentration. With the extremely low volume of solvent used, the proposed extraction method proved to be easy and simple, and adequate for high-throughput analysis at low cost. PMID:25069335

  12. [Determination of N-nitrosodimethylamine in beer by frozen zone melting liquid-liquid extraction/gas chromatography].

    PubMed

    Peng, Qiaorong; Tang, Tao; Yu, Shuxin; Sun, Yuanshe; Lei, Wu; Wang, Fengyun; Zhang, Weibing; Li, Tong

    2014-04-01

    A simple and effective sample enrichment method of frozen zone melting liquid-liquid extraction was optimized and validated for the analysis of trace N-nitrosodimethylamine (NDMA) in beer samples. The method was based on high pressure liquid-liquid extraction with a low temperature frozen step. The 90 mL beer was placed in a container with 10 mL dichloromethane. After agitation, the sample was kept in a freezer for 16 h at -19 degrees C. The organic extract was analyzed by gas chromatography with a flame ionization detector (GC-FID). The accuracy, precision, detection and quantification limits and linearity of the method were evaluated. The results showed that the calibration curve of NDMA was linear in the range of 5-200 mg/L with a good correlation coefficient (r2) of 0.999 6. The recoveries at the spiked levels of 5, 10 and 20 mg/L were 84.94%, 83.24%, 85.14% with the relative standard deviations (n = 7) of 3.06%, 3.19%, 2.63%, respectively. The ordinary extraction method of N-nitrosodimethylamine in beer includes the four steps of low-temperature distillation, liquid-liquid extraction, rotary evaporation and nitrogen blowing concentration. With the extremely low volume of solvent used, the proposed extraction method proved to be easy and simple, and adequate for high-throughput analysis at low cost.

  13. Determination of diflubenzuron and chlorbenzuron in fruits by combining acetonitrile-based extraction with dispersive liquid-liquid microextraction followed by high-performance liquid chromatography.

    PubMed

    Ruan, Chunqiang; Zhao, Xiang; Liu, Chenglan

    2015-09-01

    In this study, a simple and low-organic-solvent-consuming method combining an acetonitrile-partitioning extraction procedure followed by "quick, easy, cheap, effective, rugged and safe" cleanup with ionic-liquid-based dispersive liquid-liquid microextraction and high-performance liquid chromatography with diode array detection was developed for the determination of diflubenzuron and chlorbenzuron in grapes and pears. Ionic-liquid-based dispersive liquid-liquid microextraction was performed using the ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate as the extractive solvent and acetonitrile extract as the dispersive solvent. The main factors influencing the efficiency of the dispersive liquid-liquid microextraction were evaluated, including the extractive solvent type and volume and the dispersive solvent volume. The validation parameters indicated the suitability of the method for routine analyses of benzoylurea insecticides in a large number of samples. The relative recoveries at three spiked levels ranged between 98.6 and 109.3% with relative standard deviations of less than 5.2%. The limit of detection was 0.005 mg/kg for the two insecticides. The proposed method was successfully used for the rapid determination of diflubenzuron and chlorbenzuron residues in real fruit samples. PMID:26097091

  14. Determination of diflubenzuron and chlorbenzuron in fruits by combining acetonitrile-based extraction with dispersive liquid-liquid microextraction followed by high-performance liquid chromatography.

    PubMed

    Ruan, Chunqiang; Zhao, Xiang; Liu, Chenglan

    2015-09-01

    In this study, a simple and low-organic-solvent-consuming method combining an acetonitrile-partitioning extraction procedure followed by "quick, easy, cheap, effective, rugged and safe" cleanup with ionic-liquid-based dispersive liquid-liquid microextraction and high-performance liquid chromatography with diode array detection was developed for the determination of diflubenzuron and chlorbenzuron in grapes and pears. Ionic-liquid-based dispersive liquid-liquid microextraction was performed using the ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate as the extractive solvent and acetonitrile extract as the dispersive solvent. The main factors influencing the efficiency of the dispersive liquid-liquid microextraction were evaluated, including the extractive solvent type and volume and the dispersive solvent volume. The validation parameters indicated the suitability of the method for routine analyses of benzoylurea insecticides in a large number of samples. The relative recoveries at three spiked levels ranged between 98.6 and 109.3% with relative standard deviations of less than 5.2%. The limit of detection was 0.005 mg/kg for the two insecticides. The proposed method was successfully used for the rapid determination of diflubenzuron and chlorbenzuron residues in real fruit samples.

  15. Differentiation of mint (Mentha haplocalyx Briq.) from different regions in China using gas and liquid chromatography.

    PubMed

    Dong, Wenjiang; Ni, Yongnian; Kokot, Serge

    2015-02-01

    In this study, complex substances such as Mint (Mentha haplocalyx Briq.) samples from different growing regions in China were analyzed for phenolic compounds by high-performance liquid chromatography with diode array detection and for the volatile aroma compounds by gas chromatography with mass spectrometry. Chemometrics methods, e.g. principal component analysis, back-propagation artificial neural networks, and partial least squares discriminant analysis, were applied to resolve complex chromatographic profiles of Mint samples. A total of 49 aroma components and 23 phenolic compounds were identified in 79 Mint samples. Principal component analysis score plots from gas chromatography with mass spectrometry and high-performance liquid chromatography with diode array detection data sets showed a clear distinction among Mint from three different regions in China. Classification results showed that satisfactory performance of prediction ability for back-propagation artificial neural networks and partial least squares discriminant analysis. The major compounds that contributed to the discrimination were chlorogenic acid, unknown 3, kaempherol 7-O-rutinoside, salvianolic acid L, hesperidin, diosmetin, unknown 6 and pebrellin in Mint according to regression coefficients of the partial least squares discriminant analysis model. This study indicated that the proposed strategy could provide a simple and rapid technique to distinguish clearly complex profiles from samples such as Mint.

  16. Differentiation of mint (Mentha haplocalyx Briq.) from different regions in China using gas and liquid chromatography.

    PubMed

    Dong, Wenjiang; Ni, Yongnian; Kokot, Serge

    2015-02-01

    In this study, complex substances such as Mint (Mentha haplocalyx Briq.) samples from different growing regions in China were analyzed for phenolic compounds by high-performance liquid chromatography with diode array detection and for the volatile aroma compounds by gas chromatography with mass spectrometry. Chemometrics methods, e.g. principal component analysis, back-propagation artificial neural networks, and partial least squares discriminant analysis, were applied to resolve complex chromatographic profiles of Mint samples. A total of 49 aroma components and 23 phenolic compounds were identified in 79 Mint samples. Principal component analysis score plots from gas chromatography with mass spectrometry and high-performance liquid chromatography with diode array detection data sets showed a clear distinction among Mint from three different regions in China. Classification results showed that satisfactory performance of prediction ability for back-propagation artificial neural networks and partial least squares discriminant analysis. The major compounds that contributed to the discrimination were chlorogenic acid, unknown 3, kaempherol 7-O-rutinoside, salvianolic acid L, hesperidin, diosmetin, unknown 6 and pebrellin in Mint according to regression coefficients of the partial least squares discriminant analysis model. This study indicated that the proposed strategy could provide a simple and rapid technique to distinguish clearly complex profiles from samples such as Mint. PMID:25431171

  17. Gas chromatography on wall-coated open-tubular columns with ionic liquid stationary phases.

    PubMed

    Poole, Colin F; Lenca, Nicole

    2014-08-29

    Ionic liquids have moved from novel to practical stationary phases for gas chromatography with an increasing portfolio of applications. Ionic liquids complement conventional stationary phases because of a combination of thermophysical and solvation properties that only exist for ionic solvents. Their high thermal stability and low vapor pressure makes them suitable as polar stationary phases for separations requiring high temperatures. Ionic liquids are good solvents and can be used to expand the chemical space for separations. They are the only stationary phases with significant hydrogen-bond acidity in common use; they extend the hydrogen-bond basicity of conventional stationary phases; they are as dipolar/polarizable as the most polar conventional stationary phases; and some ionic liquids are significantly less cohesive than conventional polar stationary phases. Problems in column coating techniques and related low column performance, column activity, and stationary phase reactivity require further exploration as the reasons for these features are poorly understood at present.

  18. Analysis of limonoid glucosides from citrus by electrospray ionization liquid chromatography-mass spectrometry.

    PubMed

    Schoch, T K; Manners, G D; Hasegawa, S

    2001-03-01

    An electrospray ionization liquid chromatography-mass spectrometry (ESI-LC-MS) method for the detection and quantitation of limonoid glucosides has been developed. Negative ions [M - H(+)](-) characteristic of six limonoid glucosides can be detected and quantified from selected ion monitoring chromatograms using carminic acid as an internal standard. The described method has been applied to the analysis of limonoid glucoside content in various liquid and solid Citrus spp. samples as well as complex mixtures of partially purified limonoid glucosides. Rapid and sensitive qualitative screening of samples for limonoid glucosides can also be accomplished with slight modifications of the method. PMID:11312819

  19. Determination of preservatives in cosmetics, cleaning agents and pharmaceuticals using fast liquid chromatography.

    PubMed

    Baranowska, Irena; Wojciechowska, Iwona; Solarz, Natalia; Krutysza, Ewa

    2014-01-01

    This paper reports the development of a method for simultaneously determining five preservatives in cosmetics, cleaning agents and pharmaceuticals by fast liquid chromatography. Methylisothiazolinone, methylchloroisothiazolinone, benzyl alcohol, sodium benzoate and methylparaben were separated on a Chromolith Fast Gradient reversed-phase 18e column using gradient elution with acetonitrile and a 0.1% aqueous solution of formic acid, with a run time of 3 min. The preparation of solid and liquid samples included ultrasonic extraction with methanol with recoveries ranging from 69 to 119%. The developed method was used to analyze samples of cosmetics (66 samples), cleaning agents (five samples) and pharmaceutical industry products (17 samples).

  20. Aerosol matrix-assisted laser desorption ionization for liquid chromatography/time-of-flight mass spectrometry

    SciTech Connect

    Murray, K.K.; Lewis, T.M.; Beeson, M.D.; Russell, D.H. )

    1994-05-15

    We report the application of aerosol matrix-assisted laser desorption ionization (MALDI) to liquid chromatography/mass spectrometry (LC/MS). The aerosol MALDI experiment uses aerosol liquid introduction in conjunction with pulsed UV laser ionization to form ions from large biomolecules in solution. Mass analysis is achieved in a time-of-flight mass spectrometer. In the LC/MALDI-MS experiment, the matrix solution is combined with the column effluent in a mixing tee, LC/MALDI-MS is demonstrated for the separation of bradykinin, gramicidin S, and myoglobin. 32 refs., 8 figs., 1 tab.

  1. Kinetics of decomposition of rabeprazole sodium in aqueous solutions determined by high performance liquid chromatography.

    PubMed

    Mbah, C J

    2007-02-01

    The kinetics of decomposition of rabeprazole sodium in aqueous solutions at elevated temperatures has been investigated by high performance liquid chromatography. The reaction is found to follow first-order kinetics and the rate constant for the degradation at 25 degrees C is estimated by extrapolation. The breakdown of rabeprazole sodium is shown to be water and hydrogen ion catalysed and the effects of ionic strength and buffer concentrations to such rate studies are discussed. PMID:17341029

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

    PubMed

    Das Gupta, V

    1976-11-01

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

  3. High-performance liquid chromatography analysis of plant saponins: An update 2005-2010

    PubMed Central

    Negi, Jagmohan S.; Singh, Pramod; Pant, Geeta Joshi Nee; Rawat, M. S. M.

    2011-01-01

    Saponins are widely distributed in plant kingdom. In view of their wide range of biological activities and occurrence as complex mixtures, saponins have been purified and separated by high-performance liquid chromatography using reverse-phase columns at lower wavelength. Mostly, saponins are not detected by ultraviolet detector due to lack of chromophores. Electrospray ionization mass spectrometry, diode array detector , evaporative light scattering detection, and charged aerosols have been used for overcoming the detection problem of saponins. PMID:22303089

  4. Stability-indicating sulfa drug analysis using high-performance liquid chromatography.

    PubMed

    Umagat, H; McGarry, P F; Tscherne, R J

    1979-07-01

    Sensitive and efficient methods for sulfonamide determination as single entities and in combination with other drug substances in pharmaceutical dosage formulations were developed using high-performance liquid chromatography. These stability-indicating procedures involved a nitrile bonded phase column and nonaqueous mobile phases having diverse polarities. Sample potency was determined using peak height measurements. The methods may be used to determine trace sulfonamide quantities because detection limits are in the nanogram range.

  5. RECENT ADVANCES IN ULTRA-HIGH PERFORMANCE LIQUID CHROMATOGRAPHY FOR THE ANALYSIS OF TRADITIONAL CHINESE MEDICINE.

    PubMed

    Huang, Huilian; Liu, Min; Chen, Pei

    2014-01-01

    Traditional Chinese medicine has been widely used for the prevention and treatment of various diseases for thousands of years in China. Ultra-high performance liquid chromatography (UHPLC) is a relatively new technique offering new possibilities. This paper reviews recent developments in UHPLC in the separation and identification, fingerprinting, quantification, and metabolism of traditional Chinese medicine. Recently, the combination of UHPLC with MS has improved the efficiency of the analysis of these materials.

  6. Enantioseparation of chiral pharmaceuticals in biomedical and environmental analyses by liquid chromatography: an overview.

    PubMed

    Ribeiro, Ana R; Maia, Alexandra S; Cass, Quezia B; Tiritan, Maria E

    2014-10-01

    This review aims to present the issues associated to enantioseparation of chiral pharmaceuticals in biological and environmental matrices using chiral stationary phases (CSP). Thus, it related some enantioselective methods in liquid chromatography (LC) and compares the importance given to chiral separation in biomedical and environmental fields. For that the most used CSP, the enantioselective chromatographic methods, their advantages and drawbacks were swiftly revised and compared. The recent advances and the limitations of chiral analytical methods in LC were also discussed.

  7. Determination of dihydroergotamine in human plasma by high-performance liquid chromatography with fluorescence detection.

    PubMed

    Murday, M; McLean, A M; Slaughter, E; Couch, R A

    1999-12-10

    Dihydroergotamine, a 5-hydroxytryptamine antagonist, is used for the treatment of vascular headaches. A high-performance liquid chromatography assay with fluorescence detection is described for the determination of dihydroergotamine in plasma. The assay was validated over the concentration range 0.1-10 ng/ml plasma and applied to the analysis of plasma samples from subjects treated intramuscularly and intranasally with 2 mg of dihydroergotamine. PMID:10670731

  8. Determination of food preservatives and saccharin by high-performance liquid chromatography.

    PubMed

    Leuenberger, U; Gauch, R; Baumgartner, E

    1979-05-21

    The quantitative analysis of benzoic and sorbic acid, methyl, ethyl and propyl esters of p-hydroxybenzoic acid and saccharin in foodstuffs is described. These compounds are quantitatively extracted with disposable clean-up columns packed with Extrelut and simultaneously determined by high-performance liquid chromatography on reversed-phase columns. Complicated matrices such as cheese, cake, ketchup and chocolate were tested and recoveries were generally better than 95% in the concentration ranges normally used in the food industry. PMID:546878

  9. Isolation and Structural Elucidation of an Unknown Impurity in Prasugrel by Semi-Preparative Liquid Chromatography.

    PubMed

    Liu, Chao; Yu, Zhangxin; Wang, Fujun; Zhong, Xing; Jiang, Li; Zhang, Feifei; Tang, Yinxia; Yan, Zhaohua; Zeng, Su; Pu, Tong

    2015-08-01

    A brand-new impurity was detected by RP-HPLC in the prasugrel. The impurity was named as Impurity X. Impurity X was isolated by using semi-preparative HPLC followed by characterization using nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry. The functional mechanism of Impurity X was speculated. Impurity X could be controlled in the manufacture process of the prasugrel active pharmaceutical ingredient effectively. PMID:25644811

  10. Hydrocarbon group type determination in jet fuels by high performance liquid chromatography

    NASA Technical Reports Server (NTRS)

    Antoine, A. C.

    1977-01-01

    Thirty-two jet and diesel fuel samples of varying chemical composition and physical properties were prepared from oil shale and coal syncrudes. Hydrocarbon types in these samples were determined by a fluorescent indicator adsorption analysis, and the results from three laboratories are presented and compared. Two methods of rapid high performance liquid chromatography were used to analyze some of the samples, and these results are also presented and compared. Two samples of petroleum-based Jet A fuel are similarly analyzed.

  11. RECENT ADVANCES IN ULTRA-HIGH PERFORMANCE LIQUID CHROMATOGRAPHY FOR THE ANALYSIS OF TRADITIONAL CHINESE MEDICINE

    PubMed Central

    Huang, Huilian; Liu, Min; Chen, Pei

    2014-01-01

    Traditional Chinese medicine has been widely used for the prevention and treatment of various diseases for thousands of years in China. Ultra-high performance liquid chromatography (UHPLC) is a relatively new technique offering new possibilities. This paper reviews recent developments in UHPLC in the separation and identification, fingerprinting, quantification, and metabolism of traditional Chinese medicine. Recently, the combination of UHPLC with MS has improved the efficiency of the analysis of these materials. PMID:25045170

  12. In situ derivatization-ultrasound-assisted dispersive liquid-liquid microextraction for the determination of neurotransmitters in Parkinson's rat brain microdialysates by ultra high performance liquid chromatography-tandem mass spectrometry.

    PubMed

    He, Yongrui; Zhao, Xian-En; Zhu, Shuyun; Wei, Na; Sun, Jing; Zhou, Yubi; Liu, Shu; Liu, Zhiqiang; Chen, Guang; Suo, Yourui; You, Jinmao

    2016-08-01

    Simultaneous monitoring of several neurotransmitters (NTs) linked to Parkinson's disease (PD) has important scientific significance for PD related pathology, pharmacology and drug screening. A new simple, fast and sensitive analytical method, based on in situ derivatization-ultrasound-assisted dispersive liquid-liquid microextraction (in situ DUADLLME) in a single step, has been proposed for the quantitative determination of catecholamines and their biosynthesis precursors and metabolites in rat brain microdialysates. The method involved the rapid injection of the mixture of low toxic bromobenzene (extractant) and acetonitrile (dispersant), which containing commercial Lissamine rhodamine B sulfonyl chloride (LRSC) as derivatization reagent, into the aqueous phase of sample and buffer, and the following in situ DUADLLME procedure. After centrifugation, 50μL of the sedimented phase (bromobenzene) was directly injected for ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) detection in multiple reaction monitoring (MRM) mode. This interesting combination brought the advantages of speediness, simpleness, low matrix effects and high sensitivity in an effective way. Parameters of in situ DUADLLME and UHPLC-MS/MS conditions were all optimized in detail. The optimum conditions of in situ DUADLLME were found to be 30μL of microdialysates, 150μL of acetonitrile containing LRSC, 50μL of bromobenzene and 800μL of NaHCO3-Na2CO3 buffer (pH 10.5) for 3.0min at 37°C. Under the optimized conditions, good linearity was observed with LODs (S/N>3) and LOQs (S/N>10) of LRSC derivatized-NTs in the range of 0.002-0.004 and 0.007-0.015 nmol/L, respectively. It also brought good precision (3.2-12.8%, peak area CVs%), accuracy (94.2-108.6%), recovery (94.5-105.5%) and stability (3.8-8.1%, peak area CVs%) results. Moreover, LRSC derivatization significantly improved chromatographic resolution and MS detection sensitivity of NTs when compared with the

  13. In situ derivatization-ultrasound-assisted dispersive liquid-liquid microextraction for the determination of neurotransmitters in Parkinson's rat brain microdialysates by ultra high performance liquid chromatography-tandem mass spectrometry.

    PubMed

    He, Yongrui; Zhao, Xian-En; Zhu, Shuyun; Wei, Na; Sun, Jing; Zhou, Yubi; Liu, Shu; Liu, Zhiqiang; Chen, Guang; Suo, Yourui; You, Jinmao

    2016-08-01

    Simultaneous monitoring of several neurotransmitters (NTs) linked to Parkinson's disease (PD) has important scientific significance for PD related pathology, pharmacology and drug screening. A new simple, fast and sensitive analytical method, based on in situ derivatization-ultrasound-assisted dispersive liquid-liquid microextraction (in situ DUADLLME) in a single step, has been proposed for the quantitative determination of catecholamines and their biosynthesis precursors and metabolites in rat brain microdialysates. The method involved the rapid injection of the mixture of low toxic bromobenzene (extractant) and acetonitrile (dispersant), which containing commercial Lissamine rhodamine B sulfonyl chloride (LRSC) as derivatization reagent, into the aqueous phase of sample and buffer, and the following in situ DUADLLME procedure. After centrifugation, 50μL of the sedimented phase (bromobenzene) was directly injected for ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) detection in multiple reaction monitoring (MRM) mode. This interesting combination brought the advantages of speediness, simpleness, low matrix effects and high sensitivity in an effective way. Parameters of in situ DUADLLME and UHPLC-MS/MS conditions were all optimized in detail. The optimum conditions of in situ DUADLLME were found to be 30μL of microdialysates, 150μL of acetonitrile containing LRSC, 50μL of bromobenzene and 800μL of NaHCO3-Na2CO3 buffer (pH 10.5) for 3.0min at 37°C. Under the optimized conditions, good linearity was observed with LODs (S/N>3) and LOQs (S/N>10) of LRSC derivatized-NTs in the range of 0.002-0.004 and 0.007-0.015 nmol/L, respectively. It also brought good precision (3.2-12.8%, peak area CVs%), accuracy (94.2-108.6%), recovery (94.5-105.5%) and stability (3.8-8.1%, peak area CVs%) results. Moreover, LRSC derivatization significantly improved chromatographic resolution and MS detection sensitivity of NTs when compared with the

  14. Ionic liquid-based ultrasound-assisted dispersive liquid-liquid microextraction followed high-performance liquid chromatography for the determination of ultraviolet filters in environmental water samples.

    PubMed

    Zhang, Yufeng; Lee, Hian Kee

    2012-10-31

    In the present study, a rapid, highly efficient and environmentally friendly sample preparation method named ionic liquid-based ultrasound-assisted dispersive liquid-liquid microextraction (IL-USA-DLLME), followed by high performance liquid chromatography (HPLC) has been developed for the extraction and preconcentration of four benzophenone-type ultraviolet (UV) filters (viz. benzophenone (BP), 2-hydroxy-4-methoxybenzophenone (BP-3), ethylhexyl salicylate (EHS) and homosalate (HMS)) from three different water matrices. The procedure was based on a ternary solvent system containing tiny droplets of ionic liquid (IL) in the sample solution formed by dissolving an appropriate amount of the IL extraction solvent 1-hexyl-3-methylimidazolium tris(pentafluoroethyl)trifluorophosphate ([HMIM][FAP]) in a small amount of water-miscible dispersive solvent (methanol). An ultrasound-assisted process was applied to accelerate the formation of the fine cloudy solution, which markedly increased the extraction efficiency and reduced the equilibrium time. Various parameters that affected the extraction efficiency (such as type and volume of extraction and dispersive solvents, ionic strength, pH and extraction time) were evaluated. Under optimal conditions, the proposed method provided good enrichment factors in the range of 354-464, and good repeatability of the extractions (RSDs below 6.3%, n=5). The limits of detection were in the range of 0.2-5.0 ng mL(-1), depending on the analytes. The linearities were between 1 and 500 ng mL(-1) for BP, 5 and 500 ng mL(-1) for BP-3 and HMS and 10 and 500 ng mL(-1) for EHS. Finally, the proposed method was successfully applied to the determination of UV filters in river, swimming pool and tap water samples and acceptable relative recoveries over the range of 71.0-118.0% were obtained. PMID:23062433

  15. Determination of Niacinamide in Lotions and Creams Using Liquid-Liquid Extraction and High-Performance Liquid Chromatography

    ERIC Educational Resources Information Center

    Usher, Karyn M.; Simmons, Carolyn R.; Keating, Daniel W.; Rossi, Henry F., III

    2015-01-01

    Chemical separations are an important part of an undergraduate chemistry curriculum. Sophomore students often get experience with liquid-liquid extraction in organic chemistry classes, but liquid-liquid extraction is not as often introduced as a quantitative sample preparation method in honors general chemistry or quantitative analysis classes.…

  16. Preparative isolation and analysis of alcohol dehydrogenase inhibitors from Glycyrrhiza uralensis root using ultrafiltration combined with high-performance liquid chromatography and high-speed countercurrent chromatography.

    PubMed

    Chen, Miao; Liu, Liangliang; Chen, Xiaoqing

    2014-07-01

    A simple, rapid, and effective assay based on ultrafiltration combined with high-performance liquid chromatography and high-speed countercurrent chromatography was developed for screening and purifying alcohol dehydrogenase inhibitors from Glycyrrhiza uralensis root extract. Experiments were carried out to optimize binding conditions including alcohol dehydrogenase concentration, incubation time, temperature, and pH. By comparing the chromatograms, three compounds were found possessing alcohol dehydrogenase binding activity in Glycyrrhiza uralensis root. Under the target-guidance of ultrafiltration combined with the high-performance liquid chromatography experiment, liquiritin (1), isoliquiritin (2), and liquiritigenin (3) were separated by high-speed countercurrent chromatography using ethyl acetate/methanol/water (5:1:4) as the solvent system. The alcohol dehydrogenase inhibitory activities of these three isolated compounds were assessed; compound 2 showed strongest inhibitory activity with an IC50 of 8.95 μM. The results of the present study indicated that the combinative method using ultrafiltration, high-performance liquid chromatography and high-speed countercurrent chromatography could be widely applied for the rapid screening and isolation of enzyme inhibitors from complex mixtures.

  17. Screening for low molecular weight compounds in fish meal solubles by hydrophilic interaction liquid chromatography coupled to mass spectrometry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A simple analytical method using hydrophilic interaction liquid chromatography coupled with mass spectrometry was developed to screen for low molecular weight compounds in enzyme treated and untreated Alaskan pollock (Theragra chalcogramma) stickwater (SW) generated from processing fish meal with po...

  18. Ionic liquids for improving the extraction of NSAIDs in water samples using dispersive liquid-liquid microextraction by high performance liquid chromatography-diode array-fluorescence detection.

    PubMed

    Toledo-Neira, Carla; Álvarez-Lueje, Alejandro

    2015-03-01

    A rapid, sensitive and efficient analytical method based on the use of ionic liquids for determination of non-steroidal anti-inflammatory drugs (NSAIDs) in water samples was developed. High-performance liquid chromatography equipped with a diode array and fluorescence detector was used for quantification of ketoprofen, ibuprofen and diclofenac in tap and river water samples. This new method relies on the use of two ionic liquids with multiple functionalities: one functions as an extraction solvent (1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]), and the other changes the polarity in the aqueous medium (1-butyl-3-methylimidazolium tetrafluoroborate, ([BMIM][BF4]). Factors such as the type and volume of the ILs and dispersive solvent, sample volume, and centrifugation time were investigated and optimized. The optimized method exhibited good precision, with relative standard deviation values between 2% and 3%, for the three NSAIDs. Limits of detection achieved for all of the analytes were between 17 and 95 ng mL(-1), and the recoveries ranged from 89% to 103%. Furthermore, the enrichment factors ranged from 49 to 57. The proposed method was successfully applied to the analysis of NSAIDs in tap and river water samples.

  19. Preparation and Characterization of a Polymeric Monolithic Column for Use in High-Performance Liquid Chromatography (HPLC)

    ERIC Educational Resources Information Center

    Bindis, Michael P.; Bretz, Stacey Lowery; Danielson, Neil D.

    2011-01-01

    The high-performance liquid chromatography (HPLC) experiment, most often done in the undergraduate analytical instrumentation laboratory course, generally illustrates reversed-phase chromatography using a commercial C[subscript]18 silica column. To avoid the expense of periodic column replacement and introduce a choice of columns with different…

  20. Determination of nitrofuran metabolites in shrimp by high performance liquid chromatography with fluorescence detection and liquid chromatography-tandem mass spectrometry using a new derivatization reagent.

    PubMed

    Du, Na-Na; Chen, Ming-Ming; Sheng, Liang-Quan; Chen, Shui-Sheng; Xu, Hua-Jie; Liu, Zhao-Di; Song, Chong-Fu; Qiao, Rui

    2014-01-31

    A high performance liquid chromatography with fluorescence detection (HPLC-FLD) method for the simultaneous determination of total nitrofuran metabolite residues (furazolidone, furaltadone, nitrofurantoin, and nitrofurazone) in shrimp was developed. The method involves the acid hydrolysis of protein-bound metabolites, followed by the derivatization of the freed metabolites with the new fluorescent derivatization reagent 2-hydroxy-1-naphthaldehyde (HN) and subsequent liquid-liquid extraction (LLE). Separation is achieved on a YMC-Pack Polymer C18 column under alkaline conditions, and the high fluorescence intensity of the derivatives at an emission wavelength Em=463nm (Ex=395nm) enables, for the first time, their simultaneous determination in shrimp at concentrations as low as 1μg/kg by HPLC-FLD. The method was validated using blank shrimp fortified with all four metabolites at 0.5, 1.0 and 2.0μg/kg. Recoveries were >87% with relative standard deviations of <8.1% for all four metabolites. Furthermore, the results obtained by HPLC-FLD were in very good agreement with those obtained by LC-MS/MS analysis.

  1. Chromatographic behavior of small organic compounds in low-temperature high-performance liquid chromatography using liquid carbon dioxide as the mobile phase.

    PubMed

    Motono, Tomohiro; Nagai, Takashi; Kitagawa, Shinya; Ohtani, Hajime

    2015-07-01

    Low-temperature high-performance liquid chromatography, in which a loop injector, column, and detection cell were refrigerated at -35ºC, using liquid carbon dioxide as the mobile phase was developed. Small organic compounds (polyaromatic hydrocarbons, alkylbenzenes, and quinones) were separated by low-temperature high-performance liquid chromatography at temperatures from -35 to -5ºC. The combination of liquid carbon dioxide mobile phase with an octadecyl-silica (C18 ) column provided reversed phase mode separation, and a bare silica-gel column resulted in normal phase mode separation. In both the cases, nonlinear behavior at approximately -15ºC was found in the relationship between the temperature and the retention factors of the analytes (van't Hoff plots). In contrast to general trends in high-performance liquid chromatography, the decrease in temperature enhanced the separation efficiency of both the columns.

  2. Deformation and degradation of polymers in ultra-high-pressure liquid chromatography.

    PubMed

    Uliyanchenko, Elena; van der Wal, Sjoerd; Schoenmakers, Peter J

    2011-09-28

    Ultra-high-pressure liquid chromatography (UHPLC) using columns packed with sub-2 μm particles has great potential for separations of many types of complex samples, including polymers. However, the application of UHPLC for the analysis of polymers meets some fundamental obstacles. Small particles and narrow bore tubing in combination with high pressures generate significant shear and extensional forces in UHPLC systems, which may affect polymer chains. At high stress conditions flexible macromolecules may become extended and eventually the chemical bonds in the molecules can break. Deformation and degradation of macromolecules will affect the peak retention and the peak shape in the chromatogram, which may cause errors in the obtained results (e.g. the calculated molecular-weight distributions). In the present work we explored the limitations of UHPLC for the analysis of polymers. Degradation and deformation of macromolecules were studied by collecting and re-injecting polymer peaks and by off-line two-dimensional liquid chromatography. Polystyrene standards with molecular weight of 4 MDa and larger were found to degrade at UHPLC conditions. However, for most polymers degradation could be avoided by using low linear velocities. No degradation of 3-MDa PS (and smaller) was observed at linear velocities up to 7 mm/s. The column frits were implicated as the main sources of polymer degradation. The extent of degradation was found to depend on the type of the column and on the column history. At high flow rates degradation was observed without a column being installed. We demonstrated that polymer deformation preceded degradation. Stretched polymers eluted from the column in slalom chromatography mode (elution order opposite to that in SEC or HDC). Under certain conditions we observed co-elution of large and small PS molecules though a convolution of slalom chromatography and hydrodynamic chromatography.

  3. Identification of Clinical Isolates of Mycobacteria with Gas-Liquid Chromatography Alone

    PubMed Central

    Tisdall, Philip A.; Roberts, Glenn D.; Anhalt, John P.

    1979-01-01

    Identification of 18 mycobacterial species was performed by analysis of profiles obtained by using gas-liquid chromatography. Organisms were saponified in methanolic NaOH, and the reaction mixture was treated with BF3 in methanol and extracted with a hexane-chloroform mixture. An identification scheme was developed from 128 stock strains and tested against a collection of 79 clinical isolates. By using gas-liquid chromatographic profiles alone, 58% of specimens were correctly identified to species level, and an additional 41% were correctly identified to a group of two or three organisms. Use in a clinical laboratory over a 2-month period proved chromatography to be as accurate as and more rapid than concurrent biochemical testing. Of 81 isolates tested, 64% were identified to species level by chromatography alone. An additional 35% were differentiated to the same groups of two or three organisms as found in our analysis of stock strains. These groups consisted of: Mycobacterium tuberculosis, M. bovis, and M. xenopi; M. avium complex, M. gastri, and M. scrofulaceum; or M. fortuitum and M. chelonei. Identification to species level from these groups could usually be done by colonial morphology alone and could always be done by the addition of one selected biochemical test. This study demonstrated the practical application of gas-liquid chromatography in the identification of mycobacteria in a clinical laboratory. In particular, all strains of M. gordonae and M. kansasii were identified to species level. M. tuberculosis was definitively identified in 85% of cases. When it could not be definitely identified, the only alternatives were M. bovis and M. xenopi, both of which are rare causes of infection. PMID:118984

  4. Determination of parabens in serum by liquid chromatography-tandem mass spectrometry: Correlation with lipstick use.

    PubMed

    Tahan, Gabriella Padovani; Santos, Nayara de Kássia Souza; Albuquerque, Ana Carolina; Martins, Isarita

    2016-08-01

    Parabens are the most widely used preservative and are considered to be relatively safe compounds. However, studies have demonstrated that they may have estrogenic activity, and there is ongoing debate regarding the safety and potential cancer risk of using products containing these compounds. In the present work, liquid chromatography-tandem mass spectrometry was applied to determine methylparaben and propylparaben concentrations in serum, and the results were correlated with lipstick application. Samples were analyzed using liquid-liquid extraction, followed by liquid chromatography-tandem mass spectrometry. The validation results demonstrated the linearity of the method over a range of 1-20 ng/mL, in addition to the method's precision and accuracy. A statistically significant difference was demonstrated between serum parabens in women who used lipstick containing these substances compared with those not using this cosmetic (p = 0.0005 and 0.0016, respectively), and a strong association was observed between serum parabens and lipstick use (Spearman correlation = 0.7202).

  5. Quantitative analysis of benzodiazepines in vitreous humor by high-performance liquid chromatography

    PubMed Central

    Bazmi, Elham; Behnoush, Behnam; Akhgari, Maryam; Bahmanabadi, Leila

    2016-01-01

    Objective: Benzodiazepines are frequently screened drugs in emergency toxicology, drugs of abuse testing, and in forensic cases. As the variations of benzodiazepines concentrations in biological samples during bleeding, postmortem changes, and redistribution could be biasing forensic medicine examinations, hence selecting a suitable sample and a validated accurate method is essential for the quantitative analysis of these main drug categories. The aim of this study was to develop a valid method for the determination of four benzodiazepines (flurazepam, lorazepam, alprazolam, and diazepam) in vitreous humor using liquid–liquid extraction and high-performance liquid chromatography. Methods: Sample preparation was carried out using liquid–liquid extraction with n-hexane: ethyl acetate and subsequent detection by high-performance liquid chromatography method coupled to diode array detector. This method was applied to quantify benzodiazepines in 21 authentic vitreous humor samples. Linear curve for each drug was obtained within the range of 30–3000 ng/mL with coefficient of correlation higher than 0.99. Results: The limit of detection and quantitation were 30 and 100 ng/mL respectively for four drugs. The method showed an appropriate intra- and inter-day precision (coefficient of variation < 10%). Benzodiazepines recoveries were estimated to be over 80%. The method showed high selectivity; no additional peak due to interfering substances in samples was observed. Conclusion: The present method was selective, sensitive, accurate, and precise for the quantitative analysis of benzodiazepines in vitreous humor samples in forensic toxicology laboratory. PMID:27635251

  6. Quantitative analysis of benzodiazepines in vitreous humor by high-performance liquid chromatography

    PubMed Central

    Bazmi, Elham; Behnoush, Behnam; Akhgari, Maryam; Bahmanabadi, Leila

    2016-01-01

    Objective: Benzodiazepines are frequently screened drugs in emergency toxicology, drugs of abuse testing, and in forensic cases. As the variations of benzodiazepines concentrations in biological samples during bleeding, postmortem changes, and redistribution could be biasing forensic medicine examinations, hence selecting a suitable sample and a validated accurate method is essential for the quantitative analysis of these main drug categories. The aim of this study was to develop a valid method for the determination of four benzodiazepines (flurazepam, lorazepam, alprazolam, and diazepam) in vitreous humor using liquid–liquid extraction and high-performance liquid chromatography. Methods: Sample preparation was carried out using liquid–liquid extraction with n-hexane: ethyl acetate and subsequent detection by high-performance liquid chromatography method coupled to diode array detector. This method was applied to quantify benzodiazepines in 21 authentic vitreous humor samples. Linear curve for each drug was obtained within the range of 30–3000 ng/mL with coefficient of correlation higher than 0.99. Results: The limit of detection and quantitation were 30 and 100 ng/mL respectively for four drugs. The method showed an appropriate intra- and inter-day precision (coefficient of variation < 10%). Benzodiazepines recoveries were estimated to be over 80%. The method showed high selectivity; no additional peak due to interfering substances in samples was observed. Conclusion: The present method was selective, sensitive, accurate, and precise for the quantitative analysis of benzodiazepines in vitreous humor samples in forensic toxicology laboratory.

  7. Determination of parabens in serum by liquid chromatography-tandem mass spectrometry: Correlation with lipstick use.

    PubMed

    Tahan, Gabriella Padovani; Santos, Nayara de Kássia Souza; Albuquerque, Ana Carolina; Martins, Isarita

    2016-08-01

    Parabens are the most widely used preservative and are considered to be relatively safe compounds. However, studies have demonstrated that they may have estrogenic activity, and there is ongoing debate regarding the safety and potential cancer risk of using products containing these compounds. In the present work, liquid chromatography-tandem mass spectrometry was applied to determine methylparaben and propylparaben concentrations in serum, and the results were correlated with lipstick application. Samples were analyzed using liquid-liquid extraction, followed by liquid chromatography-tandem mass spectrometry. The validation results demonstrated the linearity of the method over a range of 1-20 ng/mL, in addition to the method's precision and accuracy. A statistically significant difference was demonstrated between serum parabens in women who used lipstick containing these substances compared with those not using this cosmetic (p = 0.0005 and 0.0016, respectively), and a strong association was observed between serum parabens and lipstick use (Spearman correlation = 0.7202). PMID:27154569

  8. Incorporation of ionic liquid into porous polymer monoliths to enhance the separation of small molecules in reversed-phase high-performance liquid chromatography.

    PubMed

    Wang, Jiafei; Bai, Ligai; Wei, Zhen; Qin, Junxiao; Ma, Yamin; Liu, Haiyan

    2015-06-01

    An ionic liquid was incorporated into the porous polymer monoliths to afford stationary phases with enhanced chromatographic performance for small molecules in reversed-phase high-performance liquid chromatography. The effect of the ionic liquid in the polymerization mixture on the performance of the monoliths was studied in detail. While monoliths without ionic liquid exhibited poor resolution and low efficiency, the addition of ionic liquid to the polymerization mixture provides highly increased resolution and high efficiency. The chromatographic performances of the monoliths were demonstrated by the separations of various small molecules including aromatic hydrocarbons, isomers, and homologues using a binary polar mobile phase. The present column efficiency reached 27 000 plates/m, which showed that the ionic liquid monoliths are alternative stationary phases in the separation of small molecules by high-performance liquid chromatography.

  9. Rapid determination of phthalate esters in alcoholic beverages by conventional ionic liquid dispersive liquid-liquid microextraction coupled with high performance liquid chromatography.

    PubMed

    Fan, Yingying; Liu, Shuhui; Xie, Qilong

    2014-02-01

    A very simple, fast and environmentally friendly sample extraction method was proposed for the analysis of phthalate esters (PAEs, di-isobutyl phthalate (DIBP), dibutylphthalate (DBP), butylbenzylphthalate (BBP) and bis(2-ethylhexyl)phthalate (DEHP)) in alcoholic beverages by using conventional ionic liquid dispersive liquid-liquid microextraction. The samples were extracted by 160 μL 1-octyl-3-methylimidazolium hexafluorophosphate in the presence of appropriate amount of ethanol and 10% (w/v) sodium chloride solution; the enriched analytes in sedimented phases were analyzed by high performance liquid chromatography-diode array detector (HPLC-DAD). Under the optimum conditions, a satisfactory linearity (in the range of 0.02-1 μg mL(-1) for white spirits and 0.01-0.5 μg mL(-1) for red wines with the correlation coefficients (r) varying from 0.9983 to 1), acceptable recovery rates (88.5-103.5% for white spirits and 91.6-104.6% for red wines), good repeatability (RSD ≤ 8.0%) and low detection limits (3.1-4.2 ng mL(-1) for white spirits and 1.5-2.2 ng mL(-1) for red wines) were obtained. The developed method was successfully applied for the determination of the four PAEs in 30 white spirits and 11 red wines collected locally, and the DBP content in 63% (19:30) white spirits exceeded the specific migration limit of 0.3 mg kg(-1) established by international regulation.

  10. Determination of a highly selective mixed-affinity sigma receptor ligand, in rat plasma by ultra performance liquid chromatography mass spectrometry and its application to a pharmacokinetic study

    PubMed Central

    Jamalapuram, Seshulatha; Vuppala, Pradeep K.; Mesangeau, Christophe; McCurdy, Christopher R.; Avery, Bonnie A.

    2014-01-01

    A selective, rapid and sensitive ultra performance liquid chromatography mass spectrometry (UPLC/MS) method was developed and validated to quantitate a highly selective mixed-affinity sigma receptor ligand, CM156 (3-(4-(4-cyclohexylpiperazin-1-yl)butyl)benzo[d] thiazole-2(3H)-thione), in rat plasma. CM156 and the internal standard (aripiprazole) were extracted from plasma samples by a single step liquid–liquid extraction using chloroform. The analysis was carried out on an ACQUITY UPLCTM BEH HILIC column (1.7 µm, 2.1 mm × 50 mm) with isocratic elution at flow rate of 0.2 mL/min using 10 mM ammonium formate in 0.1% formic acid and acetonitrile (10:90) as the mobile phase. The detection of the analyte was performed on a mass spectrometer operated in selected ion recording (SIR) mode with positive electrospray ionization (ESI). The validated analytical method resulted in a run time of 4 min and the retention times observed were 2.6 ± 0.1 and 2.1 ± 0.1 min for CM156 and the IS, respectively. The calibration curve exhibited excellent linearity over a concentration range of 5–4000 ng/mL with the lower limit of quantification of 5 ng/mL. The intra- and inter-day precision values were below 15% and accuracy ranged from −6.5% to 5.0%. The mean recovery of CM156 from plasma was 96.8%. The validated method was applied to a pilot intravenous pharmacokinetic study in rats. PMID:22406103

  11. Extraction and determination of hormones in cosmetics by homogeneous ionic liquid microextraction high-performance liquid chromatography.

    PubMed

    Kang, Mingqin; Sun, Shuo; Li, Na; Zhang, Daihui; Chen, Mingyan; Zhang, Hanqi

    2012-08-01

    The homogeneous ionic liquid microextraction was applied to the extraction of hormones from cosmetics and the hormones were determined by high-performance liquid chromatography. 1-Hexyl-3-methylimidazolium tetrafluoroborate was used as extraction solvent. Ammonium hexafluorophosphate as used as ion-pairing agent. Several experimental parameters, including the volume of 1-hexyl-3-methylimidazolium tetrafluoroborate, amount of ammonium hexafluorophosphate and sodium chloride, extraction and centrifuge time, and the pH value, were optimized. The limits of detection and quantification for the analytes ranged from 0.03 to 0.24 ng/mL and from 0.10 to 0.79 ng/mL, respectively. The precision for determining the hormones was lower than 5.2%. The proposed method was successfully developed for the determination of hormones in real cosmetic samples.

  12. Formation of iron complexs from trifluoroacetic acid based liquid chromatography mobile phases as interference ions in liquid chromatography/electrospray ionization mass spectrometric analysis

    SciTech Connect

    Shukla, Anil K.; Zhang, Rui; Orton, Daniel J.; Zhao, Rui; Clauss, Therese RW; Moore, Ronald J.; Smith, Richard D.

    2011-05-30

    Two unexpected singly charged ions at m/z 1103 and 944 have been observed in mass spectra obtained from electrospray ionization-mass spectrometric analysis of liquid chromatography effluents with mobile phases containing trifluoroacetic acid. Accurate mass measurement and tandem mass spectrometry studies revealed that these two ions are not due to any contamination from solvents and chemicals used for mobile and stationary phases or from the laboratory atmospheric environment. Instead these ions are clusters of trifluoroacetic acid formed in association with acetonitrile, water and iron from the stainless steel union used to connect the column with the electrospray tip and to apply high voltage; the molecular formulae are Fe+((OH)(H2O)2)9(CF3COOH)5 and Fe+((OH)(H2O)2)6 (CF3COOH)5.

  13. Liquid chromatography-nuclear magnetic resonance coupling as alternative to liquid chromatography-mass spectrometry hyphenations: curious option or powerful and complementary routine tool?

    PubMed

    Sturm, Sonja; Seger, Christoph

    2012-10-12

    Combining the most powerful separation techniques, i.e. liquid chromatography (LC) or capillary electrophoresis (CE) with a information rich detection system - the mass spectrometer or the nuclear magnetic resonance (NMR) spectrometer - has been pursued for more than three decades. This compilation shall provide an overview of the advantages and limitations of the LC-NMR hyphenation in the light of its most valued application-the unequivocal analyte identification. Especially the post LC trapping of analytes with an in-line solid phase extraction (SPE) device prior to transferring the analyte of interest to the NMR spectrometer (LC-SPE-NMR) proved to be a robust installation allowing a significant cut-down of the amount of analyte needed for the generation of high quality heteronuclear NMR shift correlation data. Different available technical realizations will be discussed and typical application examples from natural product research and from industrial settings will be given.

  14. Glucaminium ionic liquid-functionalized stationary phase for the separation of nucleosides in hydrophilic interaction chromatography.

    PubMed

    Jiang, Qiong; Zhang, Mingliang; Wang, Xusheng; Guo, Yong; Qiu, Hongdeng; Zhang, Shusheng

    2015-10-01

    A glucaminium-based ionic liquid stationary phase was prepared via facile epoxy-amine reaction and subsequent quaternization. Successful immobilization of glucaminium-based ionic liquid onto silica surface was validated by elemental analysis and infrared spectroscopy. The new stationary phase was evaluated for the separation of nucleosides in hydrophilic interaction liquid chromatography (HILIC). Effects of various factors, such as acetonitrile concentration, salt concentration, pH value, as well as column temperature, on the chromatographic behavior toward nucleosides were studied in detail. The results indicated that this new stationary phase can be used for separation of water-soluble polar substances in HILIC mode. The retention of solutes on the stationary phase was influenced by a mixed-mode retention mechanism with a combination of adsorptive and partitioning interactions. PMID:26231689

  15. Glucaminium ionic liquid-functionalized stationary phase for the separation of nucleosides in hydrophilic interaction chromatography.

    PubMed

    Jiang, Qiong; Zhang, Mingliang; Wang, Xusheng; Guo, Yong; Qiu, Hongdeng; Zhang, Shusheng

    2015-10-01

    A glucaminium-based ionic liquid stationary phase was prepared via facile epoxy-amine reaction and subsequent quaternization. Successful immobilization of glucaminium-based ionic liquid onto silica surface was validated by elemental analysis and infrared spectroscopy. The new stationary phase was evaluated for the separation of nucleosides in hydrophilic interaction liquid chromatography (HILIC). Effects of various factors, such as acetonitrile concentration, salt concentration, pH value, as well as column temperature, on the chromatographic behavior toward nucleosides were studied in detail. The results indicated that this new stationary phase can be used for separation of water-soluble polar substances in HILIC mode. The retention of solutes on the stationary phase was influenced by a mixed-mode retention mechanism with a combination of adsorptive and partitioning interactions.

  16. Multimycotoxin analysis in water and fish plasma by liquid chromatography-tandem mass spectrometry.

    PubMed

    Tolosa, J; Font, G; Mañes, J; Ferrer, E

    2016-02-01

    High performance liquid chromatography-mass spectrometry was used for the determination of 15 mycotoxins in water and fish plasma samples, including aflatoxins, fumonisins, ochratoxin A, sterigmatocistin, fusarenon-X and emerging Fusarium mycotoxins. In this work, dispersive liquid-liquid microextraction (DLLME) was assessed as a sample treatment for the simultaneous extraction of mycotoxins. Results showed differences in recovery assays when different extraction solvents were employed. Ethyl acetate showed better recoveries for the major part of mycotoxins analyzed, except for aflatoxins B2, G1 and G2, which showed better recoveries when employing chloroform as extractant solvent. Fumonisins and beauvericin exhibited low recoveries in both water and plasma. This method was validated according to guidelines established by European Commission and has shown to be suitable to be applied in dietary and/or toxicokinetic studies in fish where is necessary to check mycotoxin contents in rearing water and fish plasma. PMID:26694790

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

    PubMed

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

    2016-03-18

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

  18. Determination of Alternaria mycotoxins in wine and juice using ionic liquid modified countercurrent chromatography as a pretreatment method followed by high-performance liquid chromatography.

    PubMed

    Fan, Chen; Cao, Xueli; Liu, Man; Wang, Wei

    2016-03-01

    Alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA) are some of the main Alternaria mycotoxins that can be found as contaminants in food materials. The objective of this study was to develop a pretreatment method with countercurrent chromatography (CCC) for enrichment and cleanup of trace Alternaria mycotoxins in food samples prior to high-performance liquid chromatography (HPLC) analysis. An Analytical CCC instrument with a column volume 22.5mL was used, and a two-phase solvent system composed of ethyl acetate and water modified with 6% [HOOMIM][Cl] in mass to volume ratio was selected. Under the optimized CCC operation conditions, trace amounts of AOH, AME, and TeA in large volume of liquid sample were efficiently extracted and enriched in the stationary phase, and then eluted out just by reversing the stationary phase as mobile phase in the opposite flowing direction tail-to-head. The enrichment and elution strategies are unique and can be fulfilled online with high enrichment factors (87-114) and high recoveries (81.14-110.94%). The method has been successively applied to the determination of Alternaria mycotoxins in real apple juice and wine samples with the limits of detection (LOD) in the range of 0.03-0.14μgL(-1). Totally 12 wine samples and 15 apple juice samples from the local market were analyzed. The detection rate of AOH and AME in both kinds of the samples were more than 50%, while TeA was found in relatively high level of 1.75-49.61μgL(-1) in some of the apple juice samples. The proposed method is simple, rapid, and sensitive and could also be used for the analysis and monitoring of Alternaria mycotoxin in other food samples.

  19. [Qualitative and quantitative analysis of the amino acids in Rhizoma Arisaematis by ultra high performance liquid chromatography-tandem mass spectrometry and high performance liquid chromatography].

    PubMed

    Wang, Xing; Chi, Yumei; Kang, An

    2014-12-01

    A method for the identification and determination of the polar amino components without ultraviolet activity in traditional Chinese medicines was developed. With Rhizoma Arisaematis as the object of this study, using pre-column derivatization with phenyl isothiocyanate (PITC) as the derivatization reagent, compounds were separated and identified on a C18 column (100 mm x 2.1 mm, 3.5 µm) by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). A total of 20 components, including 18 amino acids and 2 amine compounds were identified. Furthermore, after the optimization of the derivatization conditions, 15 amino acids were determined by high performance liquid chromatography (HPLC) on Diamonsil C18 column (250 mm x 4.6 mm, 5 µm), detected at 254 nm and gradiently eluted by acetonitrile and 0. 05 mol/L ammonium acetate-acetic acid (pH 6. 5) as the mobile phases. The results of methodological study demonstrated that the method can meet the requirements of the determination. All calibration curves expressed good linearity: Glu, Try in the range of 2-100 mg/L, Arg in the range of 6-300 mg/L, others in the range of 0. 8-40 µg/L, with the correlation coefficients ≥ 0. 999 5. The average recovery of this method was among 95%-105% and the RSD was less than 3%. The developed method was successfully applied to quantitative determination of amino compounds in 12 batches of Rhizoma Arisaematis samples. The method is simple, sensitive, accurate, and can be used for rapid identification and determination of amino components in traditional Chinese medicines.

  20. Limonene in Arizona liquid systems used in countercurrent chromatography. I Physicochemical properties.

    PubMed

    Faure, Karine; Bouju, Elodie; Suchet, Pauline; Berthod, Alain

    2014-09-01

    Limonene is a biorenewable cycloterpene solvent derived from orange peel waste. Its potential as a "green" solvent to replace heptane was recently evaluated. Countercurrent chromatography (CCC) is a preparative separation technique using biphasic liquid systems. One liquid phase is the mobile phase; the other liquid phase is the stationary phase held in place by centrifugal fields. A particular range of special proportions of the heptane/ethyl acetate/methanol/water system is called the Arizona (AZ) liquid system when the heptane/ethyl acetate ratio is exactly the same as the methanol/water ratio. A continuous polarity decrease is obtained between the most polar A composition (ethyl acetate/water or 0/1/0/1 v/v) and the least polar Z composition (heptane/methanol or 1/0/1/0 v/v), replacing heptane by limonene and methanol by ethanol produce biphasic liquid systems much more environmentallyfriendly than the original AZ compositions. The chemical compositions of the two liquid phases of 12 AZ limonene/ethyl acetate/ethanol/water proportions were fully determined by Karl-Fisher titration of water and by gas chromatography for the organic solvents. The results were compared with the compositions of the corresponding AZ mixtures containing heptane and methanol. Significant differences in ethyl acetate and ethanol distribution between phases of the two systems with identical volume proportions were established. The ratio of the upper phase over the lower phase volumes and the phase density difference are important in CCC, there are also significant differences between the classic and "green" AZ systems that are discussed.

  1. Silica, hybrid silica, hydride silica and non-silica stationary phases for liquid chromatography.

    PubMed

    Borges, Endler M

    2015-04-01

    Free silanols on the surface of silica are the "villains", which are responsible for detrimental interactions of those compounds and the stationary phase (i.e., bad peak shape, low efficiency) as well as low thermal and chemical stability. For these reasons, we began this review describing new silica and hybrid silica stationary phases, which have reduced and/or shielded silanols. At present, in liquid chromatography for the majority of analyses, reversed-phase liquid chromatography is the separation mode of choice. However, the needs for increased selectivity and increased retention of hydrophilic bases have substantially increased the interest in hydrophilic interaction chromatography (HILIC). Therefore, stationary phases and this mode of separation are discussed. Then, non-silica stationary phases (i.e., zirconium oxide, titanium oxide, alumina and porous graphitized carbon), which afford increased thermal and chemical stability and also selectivity different from those obtained with silica and hybrid silica, are discussed. In addition, the use of these materials in HILIC is also reviewed.

  2. Analysis of trace dicyandiamide in stream water using solid phase extraction and liquid chromatography UV spectrometry.

    PubMed

    Qiu, Huidong; Sun, Dongdi; Gunatilake, Sameera R; She, Jinyan; Mlsna, Todd E

    2015-09-01

    An improved method for trace level quantification of dicyandiamide in stream water has been developed. This method includes sample pretreatment using solid phase extraction. The extraction procedure (including loading, washing, and eluting) used a flow rate of 1.0mL/min, and dicyandiamide was eluted with 20mL of a methanol/acetonitrile mixture (V/V=2:3), followed by pre-concentration using nitrogen evaporation and analysis with high performance liquid chromatography-ultraviolet spectroscopy (HPLC-UV). Sample extraction was carried out using a Waters Sep-Pak AC-2 Cartridge (with activated carbon). Separation was achieved on a ZIC(®)-Hydrophilic Interaction Liquid Chromatography (ZIC-HILIC) (50mm×2.1mm, 3.5μm) chromatography column and quantification was accomplished based on UV absorbance. A reliable linear relationship was obtained for the calibration curve using standard solutions (R(2)>0.999). Recoveries for dicyandiamide ranged from 84.6% to 96.8%, and the relative standard deviations (RSDs, n=3) were below 6.1% with a detection limit of 5.0ng/mL for stream water samples. PMID:26354690

  3. Miniaturized protein separation using a liquid chromatography column on a flexible substrate

    NASA Astrophysics Data System (ADS)

    Yang, Yongmo; Chae, Junseok

    2008-12-01

    We report a prototype protein separator that successfully miniaturizes existing technology for potential use in biocompatible health monitoring implants. The prototype is a liquid chromatography (LC) column (LC mini-column) fabricated on an inexpensive, flexible, biocompatible polydimethylsiloxane (PDMS) enclosure. The LC mini-column separates a mixture of proteins using size exclusion chromatography (SEC) with polydivinylbenzene beads (5-20 µm in diameter with 10 nm pore size). The LC mini-column is smaller than any commercially available LC column by a factor of ~11 000 and successfully separates denatured and native protein mixtures at ~71 psi of the applied fluidic pressure. Separated proteins are analyzed using NuPAGE-gel electrophoresis, high-performance liquid chromatography (HPLC) and an automated electrophoresis system. Quantitative HPLC results demonstrate successful separation based on intensity change: within 12 min, the intensity between large and small protein peaks changed by a factor of ~20. In further evaluation using the automated electrophoresis system, the plate height of the LC mini-column is between 36 µm and 100 µm. The prototype LC mini-column shows the potential for real-time health monitoring in applications that require inexpensive, flexible implant technology that can function effectively under non-laboratory conditions.

  4. Practice on high performance liquid chromatography. Applications, equipment, and quantitative analysis

    SciTech Connect

    Engelhardt, H.

    1986-01-01

    The title of this book suggests yet another general text on the principles and practice of HPLC-of which there are at least a dozen currently available. This is not the case, however. Instead, the editor has assembled a series of more specialized chapters addressed to the experienced chromatographer. That is, it is assumed that the reader has already been exposed to a more general and basic book on HPLC. Thus, there is no general discussion of the principles of separation, extracolumn effects, how an HPLC procedure is developed, column packings and techniques for reversed-phase HPLC, etc. The first five chapters of the book deal with equipment, quantitative analysis, preparative HPLC, column switching, and sample pretreatment. This paper of the book contains useful material not often seen in other books or review articles. The next two chapters deal with liquid-liquid and ion pair chromatography but ignore reversed-phase, normal-phase, and ion exchange chromatography. The final eight chapters deal with applications in different areas: inorganic samples, forensic chemistry, lipids, nucleic acid metabolites, drugs of pharmaceutical interest, amino acids, proteins, and coal and oil products. To summarize, this book is something between a basic text on HPLC and a review series such as Giddings's Advances in Chromatography. Whereas every practicing chromatographer should have a book of the first kind, libraries are the usual customers for the second kind. The author found this book useful in a variety of ways and so should many other reader.

  5. [Isolation of chemical constituents from Ziziphora clinopodioides Lam. with recycling preparative high performance liquid chromatography].

    PubMed

    Li, Guozhu; Meng, Qingyan; Luo, Bi; Ge, Zhenghong; Liu, Wenjie

    2015-01-01

    The combination of alternate recycling and direct recycling preparative liquid chromatography method was developed for the isolation of chemical constituents from Ziziphora clinopodioides Lam. The crude extract was obtained from Ziziphora clinopodioides Lam. by solvent extraction, column chromatography and reversed-phase (RP) flash chromatography. All the separations were performed with methanol and water as mobile phases and the developed recycling preparative method was used with twin RP columns switched by a two-position ten-way valve for the separation. The mobile phase was recycled in close loop with a two-position six-way valve. The fraction I and fraction II from reversed-phase flash chromatography were selected for the demonstration of separation power of the proposed protocol, and five compounds were obtained from Ziziphora clinopodioides Lam. The isolated five compounds were identified as pinocembrin-7-O-rutinoside, pinocembrin-7-O-rutinoside, acacetin-7-O-rutinoside, picein and protocatechuic acid with nuclear magnetic resonance (NMR). The experimental results showed that the developed preparation method exhibited higher separation efficiency with less mobile phase used than the reported methods, and could be expected as an effective method for the separation of complex natural products, especially the compounds with similar structures. PMID:25958674

  6. Ultrafiltration liquid chromatography combined with high-speed countercurrent chromatography for screening and isolating potential α-glucosidase and xanthine oxidase inhibitors from Cortex Phellodendri.

    PubMed

    Li, Sainan; Liu, Chunming; Guo, Liping; Zhang, Yuchi; Wang, Jing; Ma, Bing; Wang, Yueqi; Wang, Yumeng; Ren, Junqi; Yang, Xiaojing; Qin, Yao; Tang, Ying

    2014-09-01

    Cortex Phellodendri is a typical Chinese herb with a large number of alkaloids existing in all parts of it. The most common methods for screening and isolating alkaloids are mostly labor intensive and time consuming. In this study, a new assay based upon ultrafiltration liquid chromatography was developed for the rapid screening of ligands for α-glucosidase and xanthine oxidase. The C. Phellodendri extract was found to contain two alkaloids with both α-glucosidase- and xanthine oxidase binding activities and one lactone with α-glucosidase-binding activity. Subsequently, with the help of high-speed countercurrent chromatography, the specific binding ligands including palmatine, berberine, and obaculactone with purities of 97.38, 96.12, and 96.08%, respectively, were successfully separated. An optimized low-toxicity two-phase solvent system composed of ethyl acetate/n-butanol/ethanol/water (3.5:1.7:0.5:5, v/v/v/v) was used to isolate the three compounds mentioned above from C. Phellodendri. The targeted compounds were identified by liquid chromatography coupled with mass spectrometry and NMR spectroscopy. Therefore, ultrafiltration liquid chromatography combined with high-speed countercurrent chromatography is not only a powerful tool for screening and isolating α-glucosidase and xanthine oxidase inhibitors in complex samples but is also a useful platform for discovering bioactive compounds for the prevention and treatment of diabetes mellitus and gout.

  7. Improved sensitivity gas chromatography-mass spectrometry determination of parabens in waters using ionic liquids.

    PubMed

    Cacho, Juan Ignacio; Campillo, Natalia; Viñas, Pilar; Hernández-Córdoba, Manuel

    2016-01-01

    A new procedure for the introduction of ionic liquid samples in gas chromatography (GC) is proposed. This procedure, based on microvial insert thermal desorption, allows the direct analysis of the compounds preconcentrated by ionic liquid based liquid-liquid microextraction (IL-LLME) using the combination of a thermal desorption unit (TDU) and a programmed temperature vaporization injector (PTV). Two different IL-LLME methodologies, one based on the formation of a microdroplet emulsion by dispersive liquid-liquid microextraction (DLLME) and other through ultrasound-assisted emulsification microextraction (USAEME) were studied and evaluated. IL-DLLME proved advantageous and consequently, it was adopted for preconcentration purposes. This easy to apply approach was used for the determination of five parabens (methyl-, ethyl-, propyl-, butyl- and isobutyl paraben) in swimming pool waters, after in situ acetylation. The optimized conditions of TDU/PTV allowed the analytes contained in 20 µL of the enriched IL to be transferred to the capillary column. Quantification of the samples was carried out against aqueous standards, and quantification limits of between 4.3 and 8.1 ng L(-1) were obtained, depending on the compound. Concentrations of between 9 and 47 ng L(-1) for some analytes were obtained in the analysis of ten samples. PMID:26695305

  8. Analysis of Whiskey by Dispersive Liquid-Liquid Microextraction Coupled with Gas Chromatography/Mass Spectrometry: An Upper Division Analytical Chemistry Experiment Guided by Green Chemistry

    ERIC Educational Resources Information Center

    Owens, Janel E.; Zimmerman, Laura B.; Gardner, Michael A.; Lowe, Luis E.

    2016-01-01

    Analysis of whiskey samples prepared by a green microextraction technique, dispersive liquid-liquid microextraction (DLLME), before analysis by a qualitative gas chromatography-mass spectrometry (GC/MS) method, is described as a laboratory experiment for an upper division instrumental methods of analysis laboratory course. Here, aroma compounds in…

  9. Single-step conversion of cellulose to 5-hydroxymethylfurfural (HMF), a versatile platform chemical

    SciTech Connect

    Su, Yu; Brown, Heather M.; Huang, Xiwen; Zhou, Xiao Dong; Amonette, James E.; Zhang, Z. Conrad

    2009-06-20

    The ability to use cellulosic biomass as feedstock for the production of fuels and chemicals currently derived from petroleum depends critically on the development of effective low-temperature processes. While HMF, as a versatile platform chemical suitable for use in polymer synthesis or production of liquid biofuels, can currently be made from fructose and glucose, synthesis of HMF directly from raw natural cellulose represents the last major barrier toward the development of a sustainable HMF platform. Here we report an unprecedented single-step pathway that depolymerizes cellulose rapidly under mild conditions and converts the resulting glucose to hydroxymethylfurfural (HMF). A pair of metal chlorides (CuCl2 and CrCl2) dissolved in 1-ethyl-3-methylimidazolium chloride at temperatures of 80-120°C catalyzes cellulose depolymerization and the subsequent glucose conversion to HMF with 95% selectivity among recoverable products (at 56% HMF yield). Cellulose depolymerization, which can also be catalyzed by other metalchloride pairs such as CuCl2 paired with PdCl2, CrCl3, or FeCl3, occurs at a rate that is more than one order of magnitude faster than conventional acid-catalyzed hydrolysis. In contrast, single-metal chlorides at the same total loading showed low activity under similar conditions. Mechanistic studies suggest that the C2 hydrogen of the imidazolium ring is activated by the paired metal-chloride catalysts.

  10. Ionic liquid foam floatation coupled with ionic liquid dispersive liquid-liquid microextraction for the separation and determination of estrogens in water samples by high-performance liquid chromatography with fluorescence detection.

    PubMed

    Zhang, Rui; Wang, Chuanliu; Yue, Qiaohong; Zhou, Tiecheng; Li, Na; Zhang, Hanqi; Hao, Xiaoke

    2014-11-01

    An ionic liquid foam floatation coupled with ionic liquid dispersive liquid-liquid microextraction method was proposed for the extraction and concentration of 17-α-estradiol, 17-β-estradiol-benzoate, and quinestrol in environmental water samples by high-performance liquid chromatography with fluorescence detection. 1-Hexyl-3-methylimidazolium tetrafluoroborate was applied as foaming agent in the foam flotation process and dispersive solvent in microextraction. The introduction of the ion-pairing and salting-out agent NH4 PF6 was beneficial to the improvement of recoveries for the hydrophobic ionic liquid phase and analytes. Parameters of the proposed method including concentration of 1-hexyl-3-methylimidazolium tetrafluoroborate, flow rate of carrier gas, floatation time, types and concentration of ionic liquids, salt concentration in samples, extraction time, and centrifugation time were evaluated. The recoveries were between 98 and 105% with relative standard deviations lower than 7% for lake water and well water samples. The isolation of the target compounds from the water was found to be efficient, and the enrichment factors ranged from 4445 to 4632. This developing method is free of volatile organic solvents compared with regular extraction. Based on the unique properties of ionic liquids, the application of foam floatation, and dispersive liquid-liquid microextraction was widened.

  11. Rapid fractionation of collagen chains and peptides by high-performance liquid chromatography.

    PubMed

    Bateman, J F; Mascara, T; Chan, D; Cole, W G

    1986-04-01

    A strategy was developed, using a Pharmacia Fast Protein Liquid chromatography (FPLC) system, for the rapid preparation of the alpha-chains, cyanogen bromide peptides and tryptic peptides of type I collagen obtained from tissues and cultured fibroblasts. Collagen alpha-chains were prepared using a C18 PEP-RPC reverse-phase column and volatile solvents. Preliminary Superose 6 gel permeation chromatography was used to separate the crosslinked beta- and gamma-chains from the alpha-chains of tissue collagen samples. A Mono S cation-exchange column was used to resolve all of the major type I collagen cyanogen bromide peptides including the alpha 1(I)CB7 and CB8 peptides, which have not been well resolved by previously published methods. Collagen tryptic peptides were chromatographed on the PEP-RPC reverse-phase column.

  12. Identification of cosmetic dyes by ion-pair reversed-phase high-performance liquid chromatography.

    PubMed

    Gagliardi, L; Cavazzutti, G; Amato, A; Basili, A; Tonelli, D

    1987-05-22

    A method based on ion-pair reversed-phase high-performance liquid chromatography with detection at four wavelengths between 400 and 600 nm is reported for the separation and identification of the most common synthetic colour additives in cosmetic products. All the dyes generally employed in the U.S.A. and almost all those in current use in cosmetics in the European Community have been taken into account. The chromatography was performed on a C8 bonded silica packed column, with a 60-min gradient changing from 10 to 95% acetonitrile in water containing 10(-2) M sodium perchlorate (pH 3.0) as mobile phase (flow-rate 2.5 ml/min). Detection limits are in the range 20-100 ng for all dyes investigated. The method has been applied to the analysis of commercial lipsticks.

  13. Purification of dinosterol for hydrogen isotopic analysis using high-performance liquid chromatography-mass spectrometry.

    PubMed

    Smittenberg, Rienk H; Sachs, Julian P

    2007-10-26

    A semi-preparative normal-phase high-performance liquid chromatography-mass spectrometry (HPLC-MS) method is presented for the purification of various alcohol fractions from total lipid extracts derived from sediments, for the purpose of hydrogen isotopic measurement by gas chromatography-isotope ratio mass spectrometry (GC-IRMS). 4-methylsterols, including the dinoflagellate-specific marker dinosterol (4,23,24-trimethylcholestan-22-en-3beta-ol), were successfully separated from notoriously co-eluting plant-derived pentacyclic triterpenoid alcohols and alkyl alcohols. We find that substantial hydrogen isotope fractionation occurs during chromatographic separation, demonstrating the importance of recovering the entire peak when subsequent hydrogen isotope analyses are to be performed. This is the first report of such hydrogen isotopic fractionation for a natural unlabelled compound.

  14. Applications of liquid chromatography--mass spectrometry in analytical toxicology: a review.

    PubMed

    Hoja, H; Marquet, P; Verneuil, B; Lotfi, H; Pénicaut, B; Lachâtre, G

    1997-01-01

    Liquid chromatography-mass spectrometry (LC-MS), after long-term development that has introduced seven major interfacing techniques, is finally suitable for application in the field of analytical toxicology. Various compound classes can be analyzed, and sensitivities for more or less polar analytes that are as good as or better than those of gas chromatography-mass spectrometry can be obtained with modern interfaces. In addition, because ionization is often softer than classical electron impact, some LC-MS interfaces are able to handle fragile species that are otherwise not amenable to MS. This review is intended to present LC-MS to less familiarized readers and to give an extensive overview of the application of the different coupling techniques to doping agents, drugs of abuse, forensic analysis, toxic compounds of various nature, and several toxicologically relevant therapeutic drugs. Experimental parameters such as the interfaces used, ionization methods, detection limits, and experimental details for exemplary applications are given. PMID:9083829

  15. Method and apparatus for single-stepping coherence events in a multiprocessor system under software control

    DOEpatents

    Blumrich, Matthias A.; Salapura, Valentina

    2010-11-02

    An apparatus and method are disclosed for single-stepping coherence events in a multiprocessor system under software control in order to monitor the behavior of a memory coherence mechanism. Single-stepping coherence events in a multiprocessor system is made possible by adding one or more step registers. By accessing these step registers, one or more coherence requests are processed by the multiprocessor system. The step registers determine if the snoop unit will operate by proceeding in a normal execution mode, or operate in a single-step mode.

  16. Assessment of matrix effects and determination of niacin in human plasma using liquid-liquid extraction and liquid chromatography-tandem mass spectrometry.

    PubMed

    Peoples, Michael C; Halquist, Matthew S; Ismaiel, Omnia; El-Mammli, Magda Y; Shalaby, Abdalla; Karnes, H Thomas

    2008-11-01

    A simple, sensitive and rapid liquid-liquid extraction method for the analysis of nicotinic acid (niacin) and its labeled internal standard nicotinic acid-d4 (niacin-d4) in human plasma was developed and validated. The analyte and its internal standard were isolated from acidified plasma using a single liquid-liquid extraction procedure with methyl-t-butyl ether. The extracted samples were analyzed by liquid chromatography-tandem mass spectrometry in positive electrospray ionization mode with multiple reaction monitoring. The calibration curves were linear in the measured range between 5 and 1000 ng/mL and the limit of detection was calculated as 122 pg/mL. The method required 250 microL of human plasma and the total run time between injections was 3.5 min. Matrix effects were assessed by post-column infusion experiments, phospholipids monitoring and post-extraction addition experiments. The extraction of phospholipids and niacin from plasma was studied under acidic, neutral and basic conditions. Acidic conditions were optimal for both the recovery of niacin and the removal of phospholipids; the degree of matrix effects for niacin was determined to be 2.5%. It was concluded that effective removal of matrix components can overcome low recovery issues associated with liquid-liquid extractions of polar analytes.

  17. Ultrapreconcentration and determination of organophosphorus pesticides in water by solid-phase extraction combined with dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    PubMed

    Chen, Junhua; Zhou, Guangming; Deng, Yongli; Cheng, Hongmei; Shen, Jie; Gao, Yi; Peng, Guilong

    2016-01-01

    Solid-phase extraction coupled with dispersive liquid-liquid microextraction was developed as an ultra-preconcentration method for the determination of four organophosphorus pesticides (isocarbophos, parathion-methyl, triazophos and fenitrothion) in water samples. The analytes considered in this study were rapidly extracted and concentrated from large volumes of aqueous solutions (100 mL) by solid-phase extraction coupled with dispersive liquid-liquid microextraction and then analyzed using high performance liquid chromatography. Experimental variables including type and volume of elution solvent, volume and flow rate of sample solution, salt concentration, type and volume of extraction solvent and sample solution pH were investigated for the solid-phase extraction coupled with dispersive liquid-liquid microextraction with these analytes, and the best results were obtained using methanol as eluent and ethylene chloride as extraction solvent. Under the optimal conditions, an exhaustive extraction for four analytes (recoveries >86.9%) and high enrichment factors were attained. The limits of detection were between 0.021 and 0.15 μg/L. The relative standard deviations for 0.5 μg/L of the pesticides in water were in the range of 1.9-6.8% (n = 5). The proposed strategy offered the advantages of simple operation, high enrichment factor and sensitivity and was successfully applied to the determination of four organophosphorus pesticides in water samples.

  18. [Determination of endogenous steroids in urine by liquid chromatography-tandem mass spectromretry].

    PubMed

    Wang, Mengye; Xiang, Ping; Yan, Hui; Shen, Baohua; Shen, Min

    2008-01-01

    A method was developed for the determination of endogenous steroids in urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with methyltestosterone as internal standard. After enzymatic hydrolysis by beta-glucuronidase and liquid-liquid extraction, the urine sample was chromatographed on a Cosmosil C18 column with a mixture of methanol and ammonium acetate-formic acid (68:32, v/v) as mobile phase, then detected using MS/MS system with electrospray ionization (ESI) in multi-reaction monitoring (MRM) mode. The detection limits ranged from 0.01 ng/mL to 10 ng/mL. The recoveries ranged from 96.7% to 106.5%, and the intra- and inter-day precisions (measured as relative standard deviations) were less than 7% and 11%, respectively. With simple and fast sample preparation, the method was sensitive and specific for simultaneous determination of these 5 kinds of endogenous steroids in urine. The method has been successfully applied in pharmacokinetic study and is thus a potential alternative for gas chromatography-mass spectrometry (GC-MS) based procedures in routine analysis of endogenous steroids such as DHEA in human urine.

  19. Simultaneous determination of 16 synthetic colorants in hotpot condiment by high performance liquid chromatography.

    PubMed

    Tang, Bobin; Xi, Cunxian; Zou, Yun; Wang, Guomin; Li, Xianliang; Zhang, Lei; Chen, Dongdong; Zhang, Jinzhong

    2014-06-01

    A simultaneous determination method for 16 synthetic colorants in hotpot condiment was developed by high performance liquid chromatography. The samples were successively extracted with 2 mol/L carbamide solution containing 5% ammonia (dissolved in methanol) and methanol-acetone solution, and then the target analytes could be divided into two groups named as lipid-soluble and water-soluble colorants by ethyl acetate-cyclohexane with liquid-liquid extraction. The lipid-soluble and water-soluble colorants were purified by gel permeation chromatography and solid phase extraction column packed with polyamide resin, respectively. The obtained two eluates were combined, concentrated, and separated by C18 column and determined by diode array detector. Good linear relationships between peak areas and the concentrations of the synthetic colorants were obtained in the range of 0.01-50.0mg/L with correlation coefficients above 0.999 (n=10). The limits of detection and quantitation were 1-3 and 10 μg/kg for 16 synthetic colorants, respectively. The average recoveries at the spiked levels of 5, 10, 20 and 50 μg/kg were in the range of 63.2-97.1% with relative standard deviations (n=6) around 1.5-10.6%. This method is sensitive and reliable, and can be used to simultaneously determine 8 lipid-soluble and 8 water-soluble colorants in hotpot condiment. PMID:24792532

  20. Nonspecific detection of arsenic in high performance liquid chromatography: a feasibility study

    SciTech Connect

    Poulson, R.E.; Borg, H.M.

    1985-03-01

    Both gas-phase and liquid-phase systems were evaluated for their potential application in continuous on-line high performance liquid chromatography (HPLC) detection of arsenic species in oil shale byproduct waters. Previous work on related building blocks was reviewed in detail and possible systems chosen for additional laboratory evaluation. The basic system proposed includes on-line arsine generation followed by continuous gas phase detection with a thermionic ionization detector (TID) operated in the nitrogen/phosphorus mode, or liquid-phase colorimetry with silver diethyldithiocarbamate/pyridine (AgDDTC) reagent. Detection limits were calculated for arsenic based on static tests. The TID was tested in conjunction with gas chromatography of triphenylarsine, yielding 0.1 ng/min As sensitivity. The arsine AgDDTC system calculation was 50 ng/mL. Either system was deemed feasible for application to fractional ppM-level arsenic species detection on-line in HPLC effluents even though on-line sensitivities could be poorer than those calculated from the static tests by an order of magnitude. 62 refs., 4 figs., 4 tabs.

  1. Determination of maduramicin by liquid chromatography with atomic absorption spectrometric detection.

    PubMed

    Johnson, N A

    1989-01-01

    A liquid chromatograph was interfaced to an atomic absorption spectrometer for the detection and quantitation of maduramicin in feed matrixes at the 1-8 ppm level. Ionophores in general form strong 1:1 products with various metal cations, yielding complexes that are insoluble in water but very soluble in organic solvents. Maduramicin, a carboxylic, polyalcohol, polyether antibiotic, is labeled with the sodium cation and analyzed by atomic absorption spectroscopy (AAS). The lower limit of detection is approximately 100-200 ng maduramicin sodium salt. Feeds containing 1-8 ppm maduramicin are extracted with acetone, the extract is passed through an alumina column, the column is eluted with acetonitrile-water (90 + 10), and the eluate is analyzed for maduramicin by liquid chromatography-AAS after concentration and conversion of maduramicin to the sodium salt. Recoveries of maduramicin averaged 89.5%. Liquid chromatography with AAS detection has been shown to be a sensitive and highly specific technique for the determination of ionophores in general and maduramicin in particular. PMID:2708270

  2. Single-step affinity purification of enzyme biotherapeutics: a platform methodology for accelerated process development.

    PubMed

    Brower, Kevin P; Ryakala, Venkat K; Bird, Ryan; Godawat, Rahul; Riske, Frank J; Konstantinov, Konstantin; Warikoo, Veena; Gamble, Jean

    2014-01-01

    Downstream sample purification for quality attribute analysis is a significant bottleneck in process development for non-antibody biologics. Multi-step chromatography process train purifications are typically required prior to many critical analytical tests. This prerequisite leads to limited throughput, long lead times to obtain purified product, and significant resource requirements. In this work, immunoaffinity purification technology has been leveraged to achieve single-step affinity purification of two different enzyme biotherapeutics (Fabrazyme® [agalsidase beta] and Enzyme 2) with polyclonal and monoclonal antibodies, respectively, as ligands. Target molecules were rapidly isolated from cell culture harvest in sufficient purity to enable analysis of critical quality attributes (CQAs). Most importantly, this is the first study that demonstrates the application of predictive analytics techniques to predict critical quality attributes of a commercial biologic. The data obtained using the affinity columns were used to generate appropriate models to predict quality attributes that would be obtained after traditional multi-step purification trains. These models empower process development decision-making with drug substance-equivalent product quality information without generation of actual drug substance. Optimization was performed to ensure maximum target recovery and minimal target protein degradation. The methodologies developed for Fabrazyme were successfully reapplied for Enzyme 2, indicating platform opportunities. The impact of the technology is significant, including reductions in time and personnel requirements, rapid product purification, and substantially increased throughput. Applications are discussed, including upstream and downstream process development support to achieve the principles of Quality by Design (QbD) as well as integration with bioprocesses as a process analytical technology (PAT).

  3. Calculation for liquid-liquid equilibria of quaternary alkane-ethyl acetate-methanol-water systems used in counter-current chromatography.

    PubMed

    Chen, Jian; Zhao, Mengqiang; Yu, Yanmei; Li, Zongcheng

    2007-06-01

    The calculation of liquid-liquid equilibrium compositions of solvent systems is very important for the work on counter-current chromatography (CCC), especially the phase composition and volume ratio obtained from liquid-liquid equilibrium calculation. In this work, liquid-liquid equilibria of quaternary Arizona solvent systems, alkane-ethyl acetate-methanol-water, and related ternary systems are correlated and predicted using the non-random two-liquid model (NRTL). Hexane, heptane and isooctane are the used alkanes. The parameters in the model are regressed only with the special systems considered. Detailed comparison with experimental data shows that liquid-liquid equilibria of these systems can be predicted with greatly improved accuracy as compared to the group contribution method (UNIFAC).

  4. Coupling Charge Reduction Mass Spectrometry to Liquid Chromatography for Complex Mixture Analysis.

    PubMed

    Stutzman, John R; Crowe, Matthew C; Alexander, James N; Bell, Bruce M; Dunkle, Melissa N

    2016-04-01

    Electrospray ionization (ESI) of solution mixtures often generates complex mass spectra, even following liquid chromatography (LC), due to analyte multiple charging. Multiple charge state distributions can lead to isobaric interferences, mass spectral congestion, and ambiguous ion identification. As a consequence, data interpretation increases in complexity. Several charge reduction mass spectrometry (MS) approaches have been previously developed to reduce the average charge state of gaseous ions; however, all of these techniques have been restricted to direct infusion MS. In this study, synthetic polyols and surfactants separated by liquid chromatography and ionized by positive mode ESI have been subjected to polonium-210 α-particle radiation to reduce the average charge state to singly charged cations prior to mass analysis. LC/MS analysis of 5000 molecular weight poly(ethylene glycol) (PEG5000) generated an average charge state of 5.88+; whereupon, liquid chromatography/electrospray ionization/charge reduction/mass spectrometry (LC/CR/MS) analysis of PEG 5000 generated an average charge state of 1.00+. The PEG5000 results demonstrated a decrease in spectral complexity and enabled facile interpretation. Other complex solution mixtures representing specific MS challenges (i.e., competitive ionization and isobaric ion overlap) were explored and analyzed with LC/CR/MS to demonstrate the benefits of coupling LC to CR/MS. For example, polyol information related to initiator, identity/relative amount of monomer, and estimated molecular weight was characterized in random and triblock ethylene oxide/propylene oxide polyols using LC/CR/MS. LC/CR/MS is a new analytical technique for the analysis of complex mixtures. PMID:26971559

  5. Integration of electrochemistry with ultra-performance liquid chromatography/mass spectrometry.

    PubMed

    Cai, Yi; Zheng, Qiuling; Liu, Yong; Helmy, Roy; Loo, Joseph A; Chen, Hao

    2015-01-01

    This study presents the development of ultra-performance liquid chromatography (UPLC) mass spectrometry (MS) combined with electrochemistry (EC) for the first time and its application for the structural analysis of proteins/peptides that contain disulfide bonds. In our approach, a protein/peptide mixture sample undergoes a fast UPLC separation and subsequent electrochemical reduction in an electrochemical flow cell followed by online MS and tandem mass spectrometry (MS/MS) analyses. The electrochemical cell is coupled to the mass spectrometer using our recently developed desorption electrospray ionization (DESI) interface. Using this UPLC/EC/DESI-MS method, peptides that contain disulfide bonds can be differentiated from those without disulfide bonds, as the former are electroactive and reducible. MS/MS analysis of the disulfide-reduced peptide ions provides increased information on the sequence and disulfide-linkage pattern. In a reactive DESI- MS detection experiment in which a supercharging reagent was used to dope the DESI spray solvent, increased charging was obtained for the UPLC-separated proteins. Strikingly, upon online electrolytic reduction, supercharged proteins (e.g., α-lactalbumin) showed even higher charging, which will be useful in top- down protein structure MS analysis as increased charges are known to promote protein ion dissociation. Also, the separation speed and sensitivity are enhanced by approximately 1(~)2 orders of magnitude by using UPLC for the liquid chromatography (LC)/EC/MS platform, in comparison to the previously used high- performance liquid chromatography (HPLC). This UPLC/EC/DESI-MS method combines the power of fast UPLC separation, fast electrochemical conversion, and online MS structural analysis for a potentially valuable tool for proteomics research and bioanalysis.

  6. Application of a rapid and selective method for the simultaneous determination of carebastine and pseudoephedrine in human plasma by liquid chromatography-electrospray mass spectrometry for bioequivalence study in Korean subjects.

    PubMed

    Lee, Myung-Jae; Lee, Heon-Woo; Kang, Jong-Min; Seo, Ji-Hyung; Tak, Seong-Kun; Shim, Wangseob; Yim, Sung-Vin; Hong, Seung Jae; Lee, Kyung-Tae

    2010-10-01

    We describe a simple, rapid and sensitive high-performance liquid chromatography-electrospray ionization tandem mass spectrometric method that was developed for the simultaneous determination of carebastine and pseudoephedrine in human plasma using cisapride as an internal standard. Acquisition was performed in multiple-reaction monitoring mode by monitoring the transitions: m/z 500.43 > 167.09 for carebastine and m/z 166.04 > 147.88 for pseudoephedrine. The devised method involves a simple single-step liquid-liquid extraction with ethyl acetate. Chromatographic separation was performed on a C(18) reversed-phase chromatographic column at 0.2  mL/min by isocratic elution with 10  mM ammonium formate buffer-acetonitrile (30:70, v/v; adjusted to pH 3.3 with formic acid). The devised method was validated over 0.5-100  ng/mL of carebastine and 5-1000  ng/mL of pseudoephedrine with acceptable accuracy and precision, and was successfully applied to a bioequivalence study involving a single oral dose (10  mg of ebastine plus 120  mg of pseudoephedrine complex) to healthy Korean volunteers.

  7. Determination of spirocyclic tetronic/tetramic acid derivatives and neonicotinoid insecticides in fruits and vegetables by liquid chromatography and mass spectrometry after dispersive liquid-liquid microextraction.

    PubMed

    Pastor-Belda, Marta; Garrido, Isabel; Campillo, Natalia; Viñas, Pilar; Hellín, Pilar; Flores, Pilar; Fenoll, José

    2016-07-01

    Dispersive liquid-liquid microextraction was used to preconcentrate three spirocyclic tetronic/tetramic acid derivatives (spirotetramat, spiromesifen and spirodiclofen) and five neonicotinoid (thiamethoxam, chlotianidin, imidacloprid, acetamiprid and thiacloprid) insecticides previously extracted from fruit and vegetable matrices with acetonitrile. The organic enriched phase was evaporated, reconstituted in 25μL acetonitrile and analyzed by reversed-phase liquid chromatography with tandem mass spectrometry using a triple quadrupole in selected reaction monitoring mode. Enrichment factors in the 15-100 range were obtained. A matrix effect was observed, the detection limits varying between 0.025 and 0.5ngg(-1), depending on the compound and the sample matrix. The developed method was applied to the analysis of 25 samples corresponding to five different fruit and vegetable matrices. Only thiamethoxam was detected in a lemon sample at a concentration close to the quantification limit, and spiromesifen and spirotetramat at concentrations between 11.6 and 54.5ngg(-1). PMID:26920309

  8. Low density solvent-based dispersive liquid-liquid microextraction for the determination of synthetic antioxidants in beverages by high-performance liquid chromatography.

    PubMed

    Çabuk, Hasan; Köktürk, Mustafa

    2013-01-01

    A simple and efficient method was established for the determination of synthetic antioxidants in beverages by using dispersive liquid-liquid microextraction combined with high-performance liquid chromatography with ultraviolet detection. Butylated hydroxy toluene, butylated hydroxy anisole, and tert-butylhydroquinone were the antioxidants evaluated. Experimental parameters including extraction solvent, dispersive solvent, pH of sample solution, salt concentration, and extraction time were optimized. Under optimal conditions, the extraction recoveries ranged from 53 to 96%. Good linearity was observed by the square of correlation coefficients ranging from 0.9975 to 0.9997. The relative standard deviations ranged from 1.0 to 5.2% for all of the analytes. Limits of detection ranged from 0.85 to 2.73 ng mL(-1). The method was successfully applied for determination of synthetic antioxidants in undiluted beverage samples with satisfactory recoveries.

  9. Determination of alternative preservatives in cosmetic products by chromophoric derivatization followed by vortex-assisted liquid-liquid semimicroextraction and liquid chromatography.

    PubMed

    Miralles, Pablo; Vrouvaki, Ilianna; Chisvert, Alberto; Salvador, Amparo

    2016-07-01

    An analytical method for the simultaneous determination of phenethyl alcohol, methylpropanediol, phenylpropanol, caprylyl glycol, and ethylhexylglycerin, which are used as alternative preservatives in cosmetic products, has been developed. The method is based on liquid chromatography with UV spectrophotometric detection after chromophoric derivatization with benzoyl chloride and vortex-assisted liquid-liquid semimicroextraction. Different chromatographic parameters, derivatization conditions, and sample preparation variables were studied. Under optimized conditions, the limits of detection values for the analytes ranged from 0.02 to 0.06µgmL(-1). The method was validated with good recovery values (84-118%) and precision values (3.9-9.5%). It was successfully applied to 10 commercially available cosmetic samples. The good analytical features of the proposed method besides of its environmentally-friendly characteristics, make it useful to carry out the quality control of cosmetic products containing the target compounds as preservative agents.

  10. Analysis of potential migrants from plastic materials in milk by liquid chromatography-mass spectrometry with liquid-liquid extraction and low-temperature purification.

    PubMed

    Bodai, Zsolt; Szabó, Bálint Sámuel; Novák, Márton; Hámori, Susanne; Nyiri, Zoltán; Rikker, Tamás; Eke, Zsuzsanna

    2014-10-15

    A simple and fast analytical method was developed for the determination of six UV stabilizers (Cyasorb UV-1164, Tinuvin P, Tinuvin 234, Tinuvin 326, Tinuvin 327, and Tinuvin 1577) and five antioxidants (Irgafos 168, Irganox 1010, Irganox 3114, Irganox 3790, and Irganox 565) in milk. For sample preparation liquid-liquid extraction with low-temperature purification combined with centrifugation was used to remove fats, proteins, and sugars. After the cleanup step, the sample was analyzed with high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). External standard and matrix calibrations were tested. External calibration proved to be acceptable for Tinuvin P, Tinuvin 234, Tinuvin 326, Tinuvin 327, Irganox 3114, and Irganox 3790. The method was successfully validated with matrix calibration for all compounds. Method detection limits were between 0.25 and 10 μg/kg. Accuracies ranged from 93 to 109%, and intraday precisions were <13%.

  11. Determination of spirocyclic tetronic/tetramic acid derivatives and neonicotinoid insecticides in fruits and vegetables by liquid chromatography and mass spectrometry after dispersive liquid-liquid microextraction.

    PubMed

    Pastor-Belda, Marta; Garrido, Isabel; Campillo, Natalia; Viñas, Pilar; Hellín, Pilar; Flores, Pilar; Fenoll, José

    2016-07-01

    Dispersive liquid-liquid microextraction was used to preconcentrate three spirocyclic tetronic/tetramic acid derivatives (spirotetramat, spiromesifen and spirodiclofen) and five neonicotinoid (thiamethoxam, chlotianidin, imidacloprid, acetamiprid and thiacloprid) insecticides previously extracted from fruit and vegetable matrices with acetonitrile. The organic enriched phase was evaporated, reconstituted in 25μL acetonitrile and analyzed by reversed-phase liquid chromatography with tandem mass spectrometry using a triple quadrupole in selected reaction monitoring mode. Enrichment factors in the 15-100 range were obtained. A matrix effect was observed, the detection limits varying between 0.025 and 0.5ngg(-1), depending on the compound and the sample matrix. The developed method was applied to the analysis of 25 samples corresponding to five different fruit and vegetable matrices. Only thiamethoxam was detected in a lemon sample at a concentration close to the quantification limit, and spiromesifen and spirotetramat at concentrations between 11.6 and 54.5ngg(-1).

  12. Study of surface-bonded dicationic ionic liquids as stationary phases for hydrophilic interaction chromatography.

    PubMed

    Qiao, Lizhen; Li, Hua; Shan, Yuanhong; Wang, Shuangyuan; Shi, Xianzhe; Lu, Xin; Xu, Guowang

    2014-02-21

    In the present study, several geminal dicationic ionic liquids based on 1,4-bis(3-allylimidazolium)butane and 1,8-bis(3-allylimidazolium)octane in combination with different anions bromide and bis(trifluoromethanesulphonyl)imide were prepared and then bonded to the surface of 3-mercaptopropyl modified silica materials through the "thiol-ene" click chemistry as stationary phases for hydrophilic interaction chromatography (HILIC). Compared with their monocationic analogues, the dicationic ionic liquids stationary phases presented effective retention and good selectivity for typical hydrophilic compounds under HILIC mode with the column efficiency as high as 130,000 plates/m. Moreover, the influence of different alkyl chain spacer between dications and combined anions on the retention behavior and selectivity of the dicationic ionic liquids stationary phases under HILIC mode was displayed. The results indicated that the longer linkage chain would decrease the hydrophilicity and retention on the dicationic ionic liquid stationary phase, and while differently combined anions had no difference due to the exchangeability under the common HILIC mobile phase with buffer salt. Finally, the retention mechanism was investigated by evaluating the effect of chromatographic factors on retention, including the water content in the mobile phase, the mobile phase pH and buffer salt concentration. The results showed that the dicationic ionic liquids stationary phases presented a mixed-mode retention behavior with HILIC mechanism and anion exchange.

  13. High-throughput multiclass method for antibiotic residue analysis by liquid chromatography-tandem mass spectrometry.

    PubMed

    Chico, J; Rúbies, A; Centrich, F; Companyó, R; Prat, M D; Granados, M

    2008-12-12

    A simple and rapid method has been developed for the residue analysis of 39 antibiotics (tetracyclines, quinolones, penicillins, sulfonamides and macrolides) in foodstuffs of animal origin. The method combines an effective extraction technique, which uses water-methanol as extracting solvent, with ultra-high-pressure liquid chromatography-tandem mass spectrometry, allowing both confirmation and quantification in a single chromatographic run. The multiresidue method has been validated in chicken muscle matrix according to European Union Decision 2002/657/EC. It has been implemented as a routine method in a Public Health Laboratory, instead of the five plates test and LC methods previously used.

  14. Determination of amphetamine and methamphetamine in umbilical cord using liquid chromatography-tandem mass spectrometry.

    PubMed

    Jones, Joseph; Rios, Rosemarie; Jones, Mary; Lewis, Douglas; Plate, Charles

    2009-11-01

    The use of meconium as a drug-screening matrix for newborns has been the gold standard of care for the past two decades. A recent study using matched pairs of meconium and umbilical cord demonstrated a high degree of agreement. The use of liquid chromatography-tandem mass spectrometry as a means to confirm amphetamines presumptive positive umbilical cord specimens for amphetamine and methamphetamine is described here for the first time. The limit of detection for both compounds was 0.2 ng/g. The limit of quantitation for both compounds was 0.6 ng/g. The assay was linear for both compounds up to 100 ng/g.

  15. High-throughput multiclass method for antibiotic residue analysis by liquid chromatography-tandem mass spectrometry.

    PubMed

    Chico, J; Rúbies, A; Centrich, F; Companyó, R; Prat, M D; Granados, M

    2008-12-12

    A simple and rapid method has been developed for the residue analysis of 39 antibiotics (tetracyclines, quinolones, penicillins, sulfonamides and macrolides) in foodstuffs of animal origin. The method combines an effective extraction technique, which uses water-methanol as extracting solvent, with ultra-high-pressure liquid chromatography-tandem mass spectrometry, allowing both confirmation and quantification in a single chromatographic run. The multiresidue method has been validated in chicken muscle matrix according to European Union Decision 2002/657/EC. It has been implemented as a routine method in a Public Health Laboratory, instead of the five plates test and LC methods previously used. PMID:18992888

  16. Derivatization of vinyl aldehydes with anthrone prior to high-performance liquid chromatography with fluorometric detection

    SciTech Connect

    Miller, B.E.; Danielson, N.D.

    1988-04-01

    Precolumn high-performance liquid chromatography derivatization of several vinyl aldehydes, specifically acrolein, crotonaldehyde, and methacrolein, has been studied by use of the reagent anthrone. After selective condensation of these ..cap alpha..- or ..beta..-unsaturated aldehydes to form fluorescent benzanthrone derivatives, separation of these compounds was facile on a 10-cm C-18 column. Although ultraviolet detection was possible, fluorescent detection proved more versatile and sensitive. Four liquor samples were subsequently assayed for acrolein and crotonaldehyde. Linearity for these compounds in alcoholic solutions ranged from 0.02 to at least 14 ppm with detection limits reaching down to 0.005 ppm.

  17. Chiral separation of pharmaceutical compounds using electrochemically modulated liquid chromatography (EMLC)

    SciTech Connect

    Wang, S.

    1999-02-12

    This research explores the application of a new technique, termed electrochemically modulated liquid chromatography (EMLC), to the chiral separations of pharmaceutical compounds. The introduction section provides a literature review of the technique and its applications, as well as brief overview of the research described in each of the next two chapters. Chapter 2 investigates the EMLC-based enantiomeric separation of a group of chiral benzodiazepines with {beta}-cyclodextrin as a chiral mobile phase additive. Chapter 3 demonstrates the effects of several experimental parameters on the separation efficiency of drug enantiomers. The author concludes with a general summary and possible directions for future studies. Chapters 2 and 3 are processed separately.

  18. A novel arsenical in clam kidney identified by liquid chromatography/electrospray ionisation mass spectrometry.

    PubMed

    Francesconi, K A; Edmonds, J S

    2001-01-01

    Kidneys of clams of the genus Tridacna accumulate metabolic products from symbiotic unicellular algae that grow in the mantles of the clams. These metabolites include organoarsenic compounds that are biosynthesised by algae from arsenate in seawater. The arsenic compounds in aqueous extracts of the kidney of the giant clam T. derasa were investigated by liquid chromatography/electrospray ionisation mass spectrometry. About 50% of the water-soluble arsenic was present as dimethylarsinoylribosides and dimethylarsinate which are common algal metabolites. The major compound in the kidney (50% of water-soluble arsenic) was identified as a 5-dimethylarsinoyl-2,3,4-trihydroxycarboxylic acid, a new natural product.

  19. Development of an enhanced separation of erythromycin and its related substances by liquid chromatography.

    PubMed

    Deubel, A; Holzgrabe, U

    2007-01-17

    A new HPLC-UV method for the determination of the impurity profile of erythromycin is developed. In contrast to the liquid chromatography described in the European Pharmacopoeia the analysis could be performed at a temperature of 25 degrees C. Erythromycin samples were analysed on an endcapped RP phase with cyanopropyl groups on the surface using gradient elution with 32 mM potassium phosphate buffer pH 8.0 and acetonitrile/methanol (75:25). The aforementioned method shows clear improvements compared to the actual method of the European Pharmacopoeia, which is less selective and sensitive.

  20. Microwave-immobilized polybutadiene stationary phase for reversed-phase high-performance liquid chromatography.

    PubMed

    Lopes, Nilva P; Collins, Kenneth E; Jardim, Isabel C S F

    2004-03-19

    Polybutadiene (PBD) has been immobilized on high-performance liquid chromatography (HPLC) silica by microwave radiation at various power levels (52-663 W) and actuation times (3-60 min). Columns prepared from these reversed-phase HPLC materials, as well as from similar non-irradiated materials, were tested with standard sample mixtures and characterized by elemental analysis (%C) and infrared spectroscopy. A microwave irradiation of 20 min at 663 W gives a layer of immobilized PBD that presented good performance. Longer irradiation times give thicker immobilized layers having less favorable chromatographic properties.

  1. Mapping protein abundance patterns in the brain using voxelation combined with liquid chromatography and mass spectrometry

    SciTech Connect

    Petyuk, Vladislav A.; Qian, Weijun; Smith, Richard D.; Smith, Desmond J.

    2010-02-01

    Voxelation creates expression atlases by high-throughput analysis of spatially registered cubes or voxels harvested from the brain. The modality independence of voxelation allows a variety of bioanalytical techniques to be used to map abundance. Protein expression patterns in the brain can be obtained using liquid chromatography (LC) combined with mass spectrometry (MS). Here we describe the methodology of voxelation as it pertains particularly to LC-MS proteomic analysis: sample preparation, instrumental set up and analysis, peptide identification and protein relative abundance quantitation. We also briefly describe some of the advantages, limitations and insights into the brain that can be obtained using combined proteomic and transcriptomic maps

  2. Characterization of honey amino acid profiles using high-pressure liquid chromatography to control authenticity.

    PubMed

    Cotte, J F; Casabianca, H; Giroud, B; Albert, M; Lheritier, J; Grenier-Loustalot, M F

    2004-03-01

    Amino acid analysis of honey by high-performance liquid chromatography (HPLC) was used first to discriminate different botanical origins and then to combat adulteration. Pure honeys of seven selected floral varieties were examined. A principal component analysis (PCA) was carried out on the results after selection of the most discriminating parameters. Lavender honeys were thus perfectly characterized, but complete satisfaction was not obtained with the six other varieties. This method (analysis by HPLC and statistical processing by PCA) enabled us to detect the addition of sugar syrup to rape and fir honeys. PMID:14740139

  3. Quantitative Analysis of Tetramethylenedisulfotetramine ("Tetramine") Spiked into Beverages by Liquid Chromatography Tandem Mass Spectrometry with Validation by Gas Chromatography Mass Spectrometry

    SciTech Connect

    Owens, J; Hok, S; Alcaraz, A; Koester, C

    2008-11-13

    Tetramethylenedisulfotetramine, commonly known as tetramine, is a highly neurotoxic rodenticide (human oral LD{sub 50} = 0.1 mg/kg) used in hundreds of deliberate food poisoning events in China. Here we describe a method for quantitation of tetramine spiked into beverages, including milk, juice, tea, cola, and water and cleaned up by C8 solid phase extraction and liquid-liquid extraction. Quantitation by high performance liquid chromatography tandem mass spectrometry (LC/MS/MS) was based upon fragmentation of m/z 347 to m/z 268. The method was validated by gas chromatography mass spectrometry (GC/MS) operated in SIM mode for ions m/z 212, 240, and 360. The limit of quantitation was 0.10 {micro}g/mL by LC/MS/MS versus 0.15 {micro}g/mL for GC/MS. Fortifications of the beverages at 2.5 {micro}g/mL and 0.25 {micro}g/mL were recovered ranging from 73-128% by liquid-liquid extraction for GC/MS analysis, 13-96% by SPE and 10-101% by liquid-liquid extraction for LC/MS/MS analysis.

  4. Determination of three estrogens and bisphenol A by functional ionic liquid dispersive liquid-phase microextraction coupled with ultra-high performance liquid chromatography and ultraviolet detection.

    PubMed

    Jiang, Yuehuang; Tang, Tingting; Cao, Zhen; Shi, Guoyue; Zhou, Tianshu

    2015-06-01

    A hydroxyl-functionalized ionic liquid, 1-hydroxyethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, was employed in an improved dispersive liquid-phase microextraction method coupled with ultra high performance liquid chromatography for the enrichment and determination of three estrogens and bisphenol A in environmental water samples. The introduced hydroxyl group acted as the H-bond acceptor that dispersed the ionic liquid effectively in the aqueous phase without dispersive solvent or external force. Fourier transform infrared spectroscopy indicated that the hydroxyl group of the cation of the ionic liquid enhanced the combination of extractant and analytes through the formation of hydrogen bonds. The improvement of the extraction efficiency compared with that with the use of alkyl ionic liquid was proved by a comparison study. The main parameters including volume of extractant, temperature, pH, and extraction time were investigated. The calibration curves were linear in the range of 5.0-1000 μg/L for estrone, estradiol, and bisphenol A, and 10.0-1000 μg/L for estriol. The detection limits were in the range of 1.7-3.4 μg/L. The extraction efficiency was evaluated by enrichment factor that were between 85 and 129. The proposed method was proved to be simple, low cost, and environmentally friendly for the determination of the four endocrine disruptors in environmental water samples.

  5. Microfluidic aqueous two-phase extraction of bisphenol A using ionic liquid for high-performance liquid chromatography analysis.

    PubMed

    Qi, Linlin; Wang, Yunhua; Li, Yajie; Zheng, Guoxia; Li, Changping; Su, Hongzhen

    2015-05-01

    An aqueous two-phase microfluidics (ATPM) method suitable for selective extraction of bisphenol A (BPA) in aqueous samples was developed, and a functional ionic liquid of N, N, N-trioctyl ammonium propionate (TOAP) was specially employed for the formation of a parallel flow system. Based on the analytical model, we optimized the chip design into branch-connection length pattern to achieve maximum extraction efficiency (φ max) and ensure phase separation. In combining the design flexibility and ideal reaction activity of extractant (TOAP), the developed ATPM enabled a selective and effective extraction of BPA (φ max of 95% within 2 s) from phenol derivatives. Meanwhile, the total operation time and ionic liquid consumption of the microfluidic extraction were only 2.5 min and 5 μl, respectively. The ATPM can be run at normal pH and room temperature and showed no interferences from components found in tap or beach water. To be noted, this specific extraction system was applied in real water samples; the recoveries of standard addition for all water samples spiked with BPA were from 96 to 110%. Finally, successful reuse of the chip was also realized. In all cases, the developed microfluidic chip was proven to be useful as an effective and low consumption approach in extracting BPA and should be expanded as a "green" preparative method for high-performance liquid chromatography (HPLC) analysis. PMID:25796523

  6. Combining Laser Ablation/Liquid Phase Collection Surface Sampling and High-Performance Liquid Chromatography Electrospray Ionization Mass Spectrometry

    SciTech Connect

    Ovchinnikova, Olga S; Kertesz, Vilmos; Van Berkel, Gary J

    2011-01-01

    This paper describes the coupling of ambient pressure transmission geometry laser ablation with a liquid phase sample collection method for surface sampling and ionization with subsequent mass spectral analysis. A commercially available autosampler was adapted to produce a liquid droplet at the end of the syringe injection needle while in close proximity to the surface to collect the sample plume produced by laser ablation. The sample collection was followed by either flow injection or a high performance liquid chromatography (HPLC) separation of the extracted components and detection with electrospray ionization mass spectrometry (ESI-MS). To illustrate the analytical utility of this coupling, thin films of a commercial ink sample containing rhodamine 6G and of mixed isobaric rhodamine B and 6G dyes on glass microscope slides were analyzed. The flow injection and HPLC/ESI-MS analysis revealed successful laser ablation, capture and, with HPLC, the separation of the two compounds. The ablated circular area was about 70 m in diameter for these experiments. The spatial sampling resolution afforded by the laser ablation, as well as the ability to use sample processing methods like HPLC between the sample collection and ionization steps, makes this combined surface sampling/ionization technique a highly versatile analytical tool.

  7. Pressurized nano-liquid-junction interface for coupling capillary electrochromatography and nano-liquid chromatography with mass spectrometry.

    PubMed

    D'Orazio, Giovanni; Fanali, Salvatore

    2013-11-22

    A new nano-liquid-junction interface for coupling both capillary electrochromatography (CEC) or nano-liquid chromatography (nano-LC) with mass spectrometry (MS) was studied. The interface was a small T piece of polymeric material where capillary column and tip capillary were positioned at 180° while the third exit (at 90°) was occupied by a capillary delivering a liquid-assisting spray ionization for CEC experiments or by the electrode for the high voltage spray for nano-LC. Experiments were carried out analyzing mixtures of some organophosphorus pesticides (OPPs) or anti-inflammatory and related acidic drugs with MS detection in positive or negative ion mode, respectively. Analyzed OPPs compounds were baseline resolved utilizing the novel interface in both nano-LC and CEC obtaining good sensitivity and repeatability. For CEC-MS, the limits of detection ranged between 0.03 and 6.80 μg/mL and the intra-day repeatability was RSD <3.8% and <13% for the retention times and peak areas, respectively. The interface was easy to handle and good reproducibility, between 2.5 and 3.5% for the retention time and <10% for the efficiencies, was obtained when the interface was installed by two different analysts. Furthermore, it could be used for both CEC and nano-LC.

  8. Systematic evaluation of matrix effects in hydrophilic interaction chromatography versus reversed phase liquid chromatography coupled to mass spectrometry.

    PubMed

    Periat, Aurélie; Kohler, Isabelle; Thomas, Aurélien; Nicoli, Raul; Boccard, Julien; Veuthey, Jean-Luc; Schappler, Julie; Guillarme, Davy

    2016-03-25

    Reversed phase liquid chromatography (RPLC) coupled to mass spectrometry (MS) is the gold standard technique in bioanalysis. However, hydrophilic interaction chromatography (HILIC) could represent a viable alternative to RPLC for the analysis of polar and/or ionizable compounds, as it often provides higher MS sensitivity and alternative selectivity. Nevertheless, this technique can be also prone to matrix effects (ME). ME are one of the major issues in quantitative LC-MS bioanalysis. To ensure acceptable method performance (i.e., trueness and precision), a careful evaluation and minimization of ME is required. In the present study, the incidence of ME in HILIC-MS/MS and RPLC-MS/MS was compared for plasma and urine samples using two representative sets of 38 pharmaceutical compounds and 40 doping agents, respectively. The optimal generic chromatographic conditions in terms of selectivity with respect to interfering compounds were established in both chromatographic modes by testing three different stationary phases in each mode with different mobile phase pH. A second step involved the assessment of ME in RPLC and HILIC under the best generic conditions, using the post-extraction addition method. Biological samples were prepared using two different sample pre-treatments, i.e., a non-selective sample clean-up procedure (protein precipitation and simple dilution for plasma and urine samples, respectively) and a selective sample preparation, i.e., solid phase extraction for both matrices. The non-selective pretreatments led to significantly less ME in RPLC vs. HILIC conditions regardless of the matrix. On the contrary, HILIC appeared as a valuable alternative to RPLC for plasma and urine samples treated by a selective sample preparation. Indeed, in the case of selective sample preparation, the compounds influenced by ME were different in HILIC and RPLC, and lower and similar ME occurrence was generally observed in RPLC vs. HILIC for urine and plasma samples

  9. Screening antiallergic components from Carthamus tinctorius using rat basophilic leukemia 2H3 cell membrane chromatography combined with high-performance liquid chromatography and tandem mass spectrometry.

    PubMed

    Han, Shengli; Huang, Jing; Cui, Ronghua; Zhang, Tao

    2015-02-01

    Carthamus tinctorius, used in traditional Chinese medicine, has many pharmacological effects, such as anticoagulant effects, antioxidant effects, antiaging effects, regulation of gene expression, and antitumor effects. However, there is no report on the antiallergic effects of the components in C. tinctorius. In the present study, we investigated the antiallergic components of C. tinctorius and its mechanism of action. A rat basophilic leukemia 2H3/cell membrane chromatography coupled online with high-performance liquid chromatography and tandem mass spectrometry method was developed to screen antiallergic components from C. tinctorius. The screening results showed that Hydroxysafflor yellow A, from C. tinctorius, was the targeted component that retained on the rat basophilic leukemia 2H3/cell membrane chromatography column. We measured the amount of β-hexosaminidase and histamine released in mast cells and the key markers of degranulation. The release assays showed that Hydroxysafflor yellow A could attenuate the immunoglobulin E induced release of allergic cytokines without affecting cell viability from 1.0 to 50.0 μM. In conclusion, the established rat basophilic leukemia 2H3 cell membrane chromatography coupled with online high-performance liquid chromatography and tandem mass spectrometry method successfully screened and identified Hydroxysafflor yellow A from C. tinctorius as a potential antiallergic component. Pharmacological analysis elucidated that Hydroxysafflor yellow A is an effective natural component for inhibiting immunoglobulin E-antigen-mediated degranulation.

  10. Determination of nonylphenol polyethoxylates in household detergents by high-performance liquid chromatography.

    PubMed

    Cheng, Chin-Yuan; Ding, Wang-Hsien

    2002-08-30

    Preliminary survey results of the content of nonylphenol polyethoxylates (NPEOs) in various household detergents sold in Taiwan are presented. This survey was conducted to elucidate the concentration of NPEOs in household detergents and support pollution prevention and control programs. The concentrations of NPEOs in detergents and cleaners were determined by HPLC with a C8 reversed-phase column and equipped with fluorescence detection. The accuracy and precision of the method was validated and was successfully applied to determine concentrations of NPEOs in household detergents. The results show that NPEOs were detected in 41% of 90 household detergents at concentrations from 0.2 to 21%. The highest concentration of NPEOs (21%) was detected in a laundry liquid especially designed for washing socks. Reversed-phase liquid chromatography connected with electrospray mass spectrometry confirmed the results.

  11. Improving Liquid Chromatography-Mass Spectrometry Sensitivity Using a Subambient Pressure Ionization with Nanoelectrospray (SPIN) Interface

    NASA Astrophysics Data System (ADS)

    Tang, Keqi; Page, Jason S.; Marginean, Ioan; Kelly, Ryan T.; Smith, Richard D.

    2011-08-01

    In this work, the subambient pressure ionization with nanoelectrospray (SPIN) ion source and interface, which operates at ~15-30 Torr, is demonstrated to be compatible with gradient reversed-phase liquid chromatography-MS applications, exemplified here with the analysis of complex samples (a protein tryptic digest and a whole cell lysate). A low liquid chromatographic flow rate (100-400 nL/min) allowed stable electrospray to be established while avoiding electrical breakdown. Efforts to increase the operating pressure of the SPIN source relative to previously reported designs prevented solvent freezing and enhanced charged cluster/droplet desolvation. A 5- to 12-fold improvement in sensitivity relative to a conventional atmospheric pressure nanoelectrospray ionization (ESI) source was obtained for detected peptides.

  12. Novel materials and methods for solid-phase extraction and liquid chromatography

    SciTech Connect

    Ambrose, D.

    1997-06-24

    This report contains a general introduction which discusses solid-phase extraction and solid-phase micro-extraction as sample preparation techniques for high-performance liquid chromatography, which is also evaluated in the study. This report also contains the Conclusions section. Four sections have been removed and processed separately: silicalite as a sorbent for solid-phase extraction; a new, high-capacity carboxylic acid functionalized resin for solid-phase extraction; semi-micro solid-phase extraction of organic compounds from aqueous and biological samples; and the high-performance liquid chromatographic determination of drugs and metabolites in human serum and urine using direct injection and a unique molecular sieve.

  13. Saffron authentication based on liquid chromatography high resolution tandem mass spectrometry and multivariate data analysis.

    PubMed

    Rubert, Josep; Lacina, Ondrej; Zachariasova, Milena; Hajslova, Jana

    2016-08-01

    Saffron is one of the oldest and most expensive spices, which is often target of fraudulent activities. In this research, a new strategy of saffron authentication based on metabolic fingerprinting was developed. In the first phase, a solid liquid extraction procedure was optimized, the main aim was to isolate as maximal representation of small molecules contained in saffron as possible. In the second step, a detection method based on liquid chromatography coupled with high-resolution mass spectrometry was developed. Initially, principal component analysis (PCA) revealed clear differences between saffron cultivated and packaged in Spain, protected designation of origin (PDO), and saffron packaged in Spain of unknown origin, labeled Spanish saffron. Afterwards, orthogonal partial least square discriminant analysis (OPLS-DA) was favorably used to discriminate between Spanish saffron. The tentative identification of markers showed glycerophospholipids and their oxidized lipids were significant markers according to their origin. PMID:26988494

  14. Improving liquid chromatography-mass spectrometry sensitivity using a subambient pressure ionization with nanoelectrospray (SPIN) interface

    SciTech Connect

    Tang, Keqi; Page, Jason S.; Marginean, Ioan; Kelly, Ryan T.; Smith, Richard D.

    2011-04-22

    In this work the Subambient Pressure Ionization with Nanoelectrospray (SPIN) ion source and interface which operates at ~15-30 Torr is demonstrated to be compatible with gradient reversed-phase liquid chromatography-MS applications, exemplified here with the analysis of complex samples (a protein tryptic digest and a whole cell lysate). A low liquid chromatographic flow rate (100-400 nL/min) allowed stable electrospray to be established while avoiding electrical breakdown. Efforts to increase the operating pressure of the SPIN source relative to previously reported designs prevented solvent freezing and enhanced charged cluster/droplet desolvation. A 5-12-fold improvement in sensitivity relative to a conventional atmospheric pressure nanoelectrospray ionization (ESI) source was obtained for detected peptides.

  15. Determination of phosphodiesterase-5 inhibitors and analogs using high-performance liquid chromatography with ultraviolet detection.

    PubMed

    Nickum, Elisa A; Flurer, Cheryl L

    2015-01-01

    A considerable number of erectile dysfunction products, and dietary supplements suspected of containing phosphodiesterase-5 (PDE-5) inhibitors, have been analyzed by the US Food and Drug Administration. Often these samples are found to contain the approved active pharmaceutical ingredients (APIs) such as sildenafil, tadalafil or vardenafil. However, analogs of these APIs have also been identified in many samples and products containing multiple PDE-5 inhibitors have also been found. A single high-performance liquid chromatography with ultraviolet detection method has been developed for the determination of sildenafil, tadalafil, vardenafil and a number of commonly encountered analogs in pharmaceutical dosage forms and dietary supplement products, including tablets, capsules, bulk powders, troches and liquids. This method was designed as an alternative to methods developed for the determination of a single PDE-5 inhibitor. Using this protocol, 14 PDE-5 inhibitor compounds can be separated and determined in a single analysis. PMID:24668043

  16. Applying Chromatography.

    ERIC Educational Resources Information Center

    Klein, Jessie W.; Patev, Paul

    1998-01-01

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

  17. Determination of field-incurred pyrimethanil residues in persimmon (Diospyros kaki Linn) by liquid chromatography.

    PubMed

    Shim, Jae-Han; Abd El-Aty, A M; Choi, Jeong-Heui; Kang, Cheol-Ah

    2007-12-01

    Field trials have been carried out to determine the residues of the fungicide pyrimethanil in persimmon (Diospyros kaki Linn) after field treatment. Three treatments using pyrimethanil (PYR 30% WP) were carried out on persimmon trees at a recommended dose rate: the former was sprayed at two different times at 30 and 21 days prior to harvesting; the second was sprayed three times at 40, 30 and 21 days prior to harvesting; and the third was sprayed four times at 40, 30, 21 and 14 days prior to harvesting. The analysis was based on liquid-liquid extraction and solid-phase extraction followed by liquid chromatography with ultraviolet detection at 268 nm. Pyrimethanil was confirmed by high-performance liquid chromatography-mass spectrometry HPLC-MS in the selected ion monitoring mode. The method was validated with spiked fruit samples at concentrations of 0.1 and 0.4 ppm. Average recoveries (three replicates) ranged from 87.1 to 92.1% with relative standard deviations between 4.5 and 11.98%. The calculated limit of detection was 0.02 ppm and the limit of quantitation was 0.07 ppm. After two, three or four applications, pyrimethanil residues on persimmon averaged 0.44, 0.48 or 0.53 ppm, respectively; all of these values were below the maximum residue level established by the Korean Food and Drug Administration (5.0 ppm). This indicates that pyrimethanil residues declined to a level well below the maximum residue level within the 14 day period between the last application and harvesting.

  18. Improving peak shapes with counter gradients in two-dimensional high performance liquid chromatography.

    PubMed

    Stevenson, Paul G; Bassanese, Danielle N; Conlan, Xavier A; Barnett, Neil W

    2014-04-11

    To achieve the greatest peak capacity in two-dimensional high performance liquid chromatography (2D-HPLC) a gradient should be operated in both separation dimensions. However, it is known that when an injection solvent that is stronger than the initial mobile phase composition is deleterious to peak performance, thus causing problems when cutting a portion from one gradient into another. This was overcome when coupling hydrophilic interaction with reversed phase chromatography by introducing a counter gradient that changed the solvent strength of the second dimension injection. It was found that an injection solvent composition of 20% acetonitrile in water gave acceptable results in one-dimensional simulations with an initial composition of 5% acetonitrile. When this was transferred to a 2D-HPLC separation of standards it was found that a marked improvement in peak shape was gained for the moderately retained analytes (phenol and dimethyl phthalate), some improvement for the weakly retained caffeine and very little change for the strongly retained n-propylbenzene and anthracene which already displayed good chromatographic profiles. This effect was transferred when applied to a 2D-HPLC separation of a coffee extract where the indecipherable retention profile was transformed to a successful application multidimensional chromatography with peaks occupying 71% of the separation space according to the geometric approach to factor analysis. PMID:24636564

  19. Improving peak shapes with counter gradients in two-dimensional high performance liquid chromatography.

    PubMed

    Stevenson, Paul G; Bassanese, Danielle N; Conlan, Xavier A; Barnett, Neil W

    2014-04-11

    To achieve the greatest peak capacity in two-dimensional high performance liquid chromatography (2D-HPLC) a gradient should be operated in both separation dimensions. However, it is known that when an injection solvent that is stronger than the initial mobile phase composition is deleterious to peak performance, thus causing problems when cutting a portion from one gradient into another. This was overcome when coupling hydrophilic interaction with reversed phase chromatography by introducing a counter gradient that changed the solvent strength of the second dimension injection. It was found that an injection solvent composition of 20% acetonitrile in water gave acceptable results in one-dimensional simulations with an initial composition of 5% acetonitrile. When this was transferred to a 2D-HPLC separation of standards it was found that a marked improvement in peak shape was gained for the moderately retained analytes (phenol and dimethyl phthalate), some improvement for the weakly retained caffeine and very little change for the strongly retained n-propylbenzene and anthracene which already displayed good chromatographic profiles. This effect was transferred when applied to a 2D-HPLC separation of a coffee extract where the indecipherable retention profile was transformed to a successful application multidimensional chromatography with peaks occupying 71% of the separation space according to the geometric approach to factor analysis.

  20. Enantiomeric separation of organophosphorus pesticides by high-performance liquid chromatography, gas chromatography and capillary electrophoresis and their applications to environmental fate and toxicity assays.

    PubMed

    Li, Ling; Zhou, Shanshan; Jin, Lixia; Zhang, Cheng; Liu, Weiping

    2010-05-15

    In recent years, the continuous evolution of the field of stereochemistry has produced a heightened awareness of the applications of pure enantiomers of agrochemicals. This review describes reports of the enantiomeric separation of commercial organophosphorus pesticides (OPs) and the applications of these methods to research on the enantioselectivity of the toxicity and environmental fate of these compounds. Chiral OPs can be analysed by high-performance liquid chromatography (HPLC), gas chromatography (GC), and capillary electrophoresis (CE). These different separation techniques for OP enantiomers are briefly discussed, and their applications are presented.

  1. Protein quantitation using Ru-NHS ester tagging and isotope dilution high-pressure liquid chromatography-inductively coupled plasma mass spectrometry determination.

    PubMed

    Liu, Rui; Lv, Yi; Hou, Xiandeng; Yang, Lu; Mester, Zoltan

    2012-03-20

    An accurate, simple, and sensitive method for the direct determination of proteins by nonspecies specific isotope dilution and external calibration high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICPMS) is described. The labeling of myoglobin (17 kDa), transferrin (77 kDa), and thyroglobulin (670 kDa) proteins was accomplished in a single-step reaction with a commercially available bis(2,2'-bipyridine)-4'-methyl-4-carboxybipyridine-ruthenium N-succinimidyl ester-bis(hexafluorophosphate) (Ru-NHS ester). Using excess amounts of Ru-NHS ester compared to the protein concentration at optimized labeling conditions, constant ratios for Ru to proteins were obtained. Bioconjugate solutions containing both labeled and unlabeled proteins as well as excess Ru-NHS ester reagent were injected onto a size exclusion HPLC column for separation and ICPMS detection without any further treatment. A (99)Ru enriched spike was used for nonspecies specific ID calibration. The accuracy of the method was confirmed at various concentration levels. An average recovery of 100% ± 3% (1 standard deviation (SD), n = 9) was obtained with a typical precision of better than 5% RSD at 100 μg mL(-1) for nonspecies specific ID. Detection limits (3SD) of 1.6, 3.2, and 7.0 fmol estimated from three procedure blanks were obtained for myoglobin, transferrin, and thyroglobulin, respectively. These detection limits are suitable for the direct determination of intact proteins at trace levels. For simplicity, external calibration was also tested. Good linear correlation coefficients, 0.9901, 0.9921, and 0.9980 for myoglobin, transferrin, and thyroglobulin, respectively, were obtained. The measured concentrations of proteins in a solution were in good agreement with their volumetrically prepared values. To the best of our knowledge, this is the first application of nonspecies specific ID for the accurate and direct determination of proteins using a Ru-NHS ester

  2. Matrix solid-phase dispersion followed by liquid chromatography tandem mass spectrometry for the determination of selective ciclooxygenase-2 inhibitors in sewage sludge samples.

    PubMed

    Triñanes, Sara; Casais, M Carmen; Mejuto, M Carmen; Cela, Rafael

    2016-09-01

    A straightforward single-step extraction method based on matrix solid-phase dispersion (MSPD), followed by high-performance liquid chromatography with hybrid quadrupole time of flight mass spectrometry (LC-QTOF-MS), was developed and optimized to determine five non-steroidal anti-inflammatory drugs (Valdecoxib, Etoricoxib, Parecoxib, Celecoxib and 2,5-Dimethylcelecoxib) in sewage sludge samples. The influence of different operational parameters on the extraction efficiency a well as in the matrix effects of the produced extracts was evaluated in detail. Under final working conditions, freeze dried samples (0.2g) were first soaked with 100μL of aqueous potassium hydroxide solution (60%, w/v), mixed with 1g of anhydrous sodium sulfate and dispersed with 1g of Florisil. This blend was transferred to the top of a polypropylene column cartridge containing 3g of silica. Analytes were recovered using 15mL of hexane/acetone (1:2, v/v) mixture. The extracts were concentrated by evaporation and reconstituted with 1mL of methanol/water (1:1, v/v), filtered and injected in the LC system. Quantification limits from 0.005 and 0.05ngg(-1) and absolute recoveries between 86 and 105% were achieved. Results indicated the presence of two of the targeted COXIBs in real samples of sewage sludge, the highest average concentration (22ngg(-1)) corresponding to celecoxib. Moreover, the screening capabilities of the LC-QTOF-MS system demonstrated that the developed MSPD extraction procedure might be useful for the selective extraction of some other pharmaceuticals (e.g. amiodarone and their metabolite N-desethylamiodarone, miconazole, clotrimazole and ketoprofen) from sludge samples. PMID:27521258

  3. Matrix solid-phase dispersion followed by liquid chromatography tandem mass spectrometry for the determination of selective ciclooxygenase-2 inhibitors in sewage sludge samples.

    PubMed

    Triñanes, Sara; Casais, M Carmen; Mejuto, M Carmen; Cela, Rafael

    2016-09-01

    A straightforward single-step extraction method based on matrix solid-phase dispersion (MSPD), followed by high-performance liquid chromatography with hybrid quadrupole time of flight mass spectrometry (LC-QTOF-MS), was developed and optimized to determine five non-steroidal anti-inflammatory drugs (Valdecoxib, Etoricoxib, Parecoxib, Celecoxib and 2,5-Dimethylcelecoxib) in sewage sludge samples. The influence of different operational parameters on the extraction efficiency a well as in the matrix effects of the produced extracts was evaluated in detail. Under final working conditions, freeze dried samples (0.2g) were first soaked with 100μL of aqueous potassium hydroxide solution (60%, w/v), mixed with 1g of anhydrous sodium sulfate and dispersed with 1g of Florisil. This blend was transferred to the top of a polypropylene column cartridge containing 3g of silica. Analytes were recovered using 15mL of hexane/acetone (1:2, v/v) mixture. The extracts were concentrated by evaporation and reconstituted with 1mL of methanol/water (1:1, v/v), filtered and injected in the LC system. Quantification limits from 0.005 and 0.05ngg(-1) and absolute recoveries between 86 and 105% were achieved. Results indicated the presence of two of the targeted COXIBs in real samples of sewage sludge, the highest average concentration (22ngg(-1)) corresponding to celecoxib. Moreover, the screening capabilities of the LC-QTOF-MS system demonstrated that the developed MSPD extraction procedure might be useful for the selective extraction of some other pharmaceuticals (e.g. amiodarone and their metabolite N-desethylamiodarone, miconazole, clotrimazole and ketoprofen) from sludge samples.

  4. Flumethrin residue levels in honey from apiaries of China by high-performance liquid chromatography.

    PubMed

    Yu, L S; Liu, F; Wu, H; Tan, H R; Ruan, X C; Chen, Y; Chao, Z

    2015-01-01

    A method for detection of flumethrin residue in honey by high-performance liquid chromatography was established. After n-hexane-dichloromethane (4:6, vol/vol) extraction, the honey samples were concentrated by rotary evaporation, purified by an Oasis HLB solid-phase extraction column, and detected using a UV detector at 267 nm. The interference of the matrix was greatly reduced by optimizing pretreatment conditions; thus, the minimal detection limit of cyhalothrin was 0.005 mg/kg, the average recovery was 80.8 to 96.8%, and the coefficient of variation was 0.6 to 1.5%. The precision and reproducibility of this method was suitable and applicable for detecting flumethrin residue in honey. With this method, 135 honey samples from seven locations in the People's Republic of China were tested; 77 samples tested positive for flumethrin residue, resulting in a detection rate of 75.3%. Samples from the Guangdong province had the highest flumethrin residue level (0.122 mg/kg) of the locations tested. On the basis of analytical validation, the high-performance liquid chromatography has been shown to be a promising alternative for the analysis of flumethrin residue in honey samples. PMID:25581190

  5. Separation of tryptophan enantiomers by ligand-exchange chromatography with novel chiral ionic liquids ligand.

    PubMed

    Qing, Haiqun; Jiang, Xinyu; Yu, Jingang

    2014-03-01

    Chiral ionic liquids (CILs) with amino acids as cations have been applied as novel chiral ligands coordinated with Cu(2+) to separate tryptophan enantiomers in ligand exchange chromatography. Four kinds of amino acid ionic liquids, including [L-Pro][CF3COO], [L-Pro][NO3], [L-Pro]2[SO4], and [L-Phe][CF3COO] were successfully synthesized and used for separation of tryptophan enantiomers. To optimize the separation conditions, [L-Pro][CF3COO] was selected as the model ligand. Some factors influencing the efficiency of chiral separation, such as copper ion concentration, CILs concentration, methanol ratio (methanol/H2O, v/v), and pH, were investigated. The obtained optimal separation conditions were as follows: 8.0 mmol/L Cu(OAc)2, 4.0 mmol/L [L-Pro][CF3COO], and 20% (v/v) methanol at pH 3.6. Under the optimum conditions, acceptable enantioseparation of tryptophan enantiomers could be observed with a resolution of 1.89. The results demonstrate the good applicability of CILs with amino acids as cations for chiral separation. Furthermore, a comparative study was also conducted for exploring the mechanism of the CILs as new ligands in ligand exchange chromatography.

  6. Constant pressure mode extended simple gradient liquid chromatography system for micro and nanocolumns.

    PubMed

    Šesták, Jozef; Kahle, Vladislav

    2014-07-11

    Performing gradient liquid chromatography at constant pressure instead of constant flow rate has serious potential for shortening the analysis time and increasing the productivity of HPLC instruments that use gradient methods. However, in the constant pressure mode the decreasing column permeability during a long period of time negatively affects the repeatability of retention time. Thus a volume-based approach, in which the detector signal is plotted as a function of retention volume, must be taken into consideration. Traditional HPLC equipment, however, requires quite complex hardware and software modifications in order to work at constant pressure and in the volume-based mode. In this short communication, a low cost and easily feasible pressure-controlled extension of the previously described simple gradient liquid chromatography platform is proposed. A test mixture of four nitro esters was separated by 10-60% (v/v) acetone/water gradient and a high repeatability of retention volumes at 20MPa (RSD less than 0.45%) was realized. Separations were also performed at different values of pressure (20, 25, and 31MPa), and only small variations of the retention volumes (up to 0.8%) were observed. In this particular case, the gain in the analysis speed of 7% compared to the constant flow mode was realized at a constant pressure.

  7. Stationary phase modulation in liquid chromatography through the serial coupling of columns: A review.

    PubMed

    Alvarez-Segura, T; Torres-Lapasió, J R; Ortiz-Bolsico, C; García-Alvarez-Coque, M C

    2016-06-01

    Liquid chromatography with single columns often does not succeed in the analysis of complex samples, in terms of resolution and analysis time. A relatively simple solution to enhance chromatographic resolution is the modulation of the stationary phase through the serial coupling of columns. This can be implemented with any type of column using compatible elution conditions and conventional instruments. This review describes the key features of column coupling and published procedures, where two or more columns were coupled in series to solve separation problems. In all reports, the authors could not resolve their samples with single columns, whereas significant enhancement in chromatographic performance was obtained when the columns were combined. Particularly interesting is the reduction in the analysis time in the isocratic mode, which alleviates the "general elution problem" of liquid chromatography, and may represent a stimulus for the proposal of new procedures, especially in combination with mass spectrometric, electrochemical and refractometric detection. Developments proposed to make the serial coupling of columns useful in routine and research laboratories are outlined, including optimisation strategies that facilitate the selection of the appropriate column combination and elution conditions (solvent content, flow rate or temperature) in both isocratic and gradient modes. The availability of zero dead volume couplers, able to connect standard columns, and the commercialisation of short columns with multiple lengths, have expanded the possibilities of success. PMID:27155298

  8. Determination of bromate in drinking water by ultraperformance liquid chromatography-tandem mass spectrometry.

    PubMed

    Alsohaimi, Ibrahim Hotan; Alothman, Zeid Abdullah; Khan, Mohammad Rizwan; Abdalla, Mohammad Abulhassan; Busquets, Rosa; Alomary, Ahmad Khodran

    2012-10-01

    Bromate is a byproduct formed as a result of disinfection of bromide-containing source water with ozone or hypochlorite. The International Agency for Research on Cancer has recognized bromate as a possible human carcinogen, thus it is essential to determine in drinking water. Present work highlights a development of sensitive and fast analytical method for bromate determination in drinking water by using ultraperformance liquid chromatography-tandem mass spectrometry. The quality parameters of the developed method were established, obtaining very low limit of detection (0.01 ng/mL), repeatability and reproducibility have been found to be less than 3% in terms of relative standard deviation when analyzing a bromate standard at 0.05 μg/mL with 0.4 min analysis time. Developed method was applied for the analysis of metropolitan and bottled water from Saudi Arabia; 22 samples have been analyzed. Bromate was detected in the metropolitan water samples (from desalinization source) at concentrations ranging between 3.43 and 75.04 ng/mL and in the bottled water samples at concentrations ranging between 2.07 and 21.90 ng/mL. Moreover, in comparison to established analytical methods such as liquid chromatography-tandem mass spectrometry, the proposed method was found to be very sensitive, selective and rapid for the routine analysis of bromate at low level in drinking water.

  9. Determination of ochratoxin A in wine by immunoaffinity cleanup and liquid chromatography tandem mass spectrometry.

    PubMed

    Noba, Shigekuni; Omote, Masayuki; Kitagawa, Yasushi; Mochizuki, Naoki

    2008-05-01

    A simple and accurate method has been developed for determining ochratoxin A (OTA), using an immunoaffinity column for cleanup and liquid chromatography-tandem mass spectrometry for identification and quantification. Wine samples were diluted with a solution containing polyethylene glycol 8000 and sodium hydrogen carbonate, filtered through a glass microfiber filter, and cleaned up on an immunoaffinity column. OTA was then eluted with methanol-acetic acid (98:2) and analyzed by liquid chromatography-tandem mass spectrometry. The average recoveries of OTA from red and white wines were 95 and 96.7% (spiked OTA level was 0.05 ng/ml) and repeatabilities (relative standard deviation) were 3.8 and 2.4%, respectively. The detection limit was 0.0003 ng/ml based on the signal-to-noise ratio in wine of 3:1. Analysis of 74 samples of domestic and imported wines showed OTA levels ranging from < 0.0003 to 0.82 ng/ml, with an incidence of contamination of 92.1% for red wines, and < 0.0003 to 0.51 ng/ml, with an incidence of contamination of 77.8% for white wines. These detection rates were higher than those rates of past reports of OTA contamination in wine, due to the high sensitivity of this method. However, all samples analyzed in this study complied with European Union regulations. It is concluded that this method is a useful tool for the quality assurance of wine.

  10. Determination of phthalate esters in soil samples by microwave assisted extraction and high performance liquid chromatography.

    PubMed

    Liang, Pei; Zhang, Linlin; Peng, Lili; Li, Qian; Zhao, Ehong

    2010-08-01

    A method was developed for the determination of phthalate esters (dimethyl phthalate, diethyl phthalate, benzyl butyl phthalate, di-n-butyl phthalate, di-n-octyl phthalate and di-(2-ethylhextyl) phthalate) in soil samples. The method was based on microwave-assisted extraction of soil using acetonitrile as extractant. Phthalate esters in the extract were determined by high performance liquid chromatography with variable wavelength detector. Microwave-assisted extraction operational parameters, such as the solvent type, extraction temperature and time, were studied and optimized. Under the resultant conditions, recoveries of phthalate esters from spiked soil samples were in the range from 84 to 115% for two different spiking levels (0.1 and 0.5 microg g(-1)), and relative standard deviations of the recoveries were below 8%. The limits of detection ranged from 1.24 to 3.15 microg L(-1). The method did not require clean-up or preconcentration steps. The obtained results showed that microwave-assisted extraction combined with high performance liquid chromatography was a fast and simple method for the determination of phthalate esters in soil samples.

  11. Simultaneous determination of airborne carbamates in workplace by high performance liquid chromatography with fluorescence detection.

    PubMed

    Li, Hong-Ping; Li, Jen-Hou; Li, Gwo-Chen; Jen, Jen-Fon

    2004-06-17

    A high performance liquid chromatography with fluorescence (HPLC-F) detector was examined to simultaneous determination of airborne carbamates in the workplace of manufactory. The OVS-2 air sampling tube filled with glass fiber filter or quartz fiber and combined filter/XAD-2 were evaluated to collect nine commonly used carbamates (Carbofuran, Isoprocarb, Methomyl, Metolcarb, Thiodicarb, Carbaryl, Oxamyl, Methiocarb, and Prpoxur) from the air of manufactory in high humidity country. After being extracted with acetonitrile from sampling tubes, the carbamates were determined by high performance liquid chromatography with fluorescence detection posterior to on-line derivatization. The collection of carbamates and the recovery of extraction from glass wool fiber in several concentration levels, and from quartz filter were evaluated. The storage stability of carbamates was also tested. Results indicated that the HPLC-fluorescence method offers satisfactory resolution and sensitivity in carbamate analysis. With the glass fiber filter and combined filter/XAD-2, the Carbofuran, Isoprocarb, Methomyl, Metolcarb, and Thiodicarb were stable for a 28-day storage test, Carbaryl and Oxamyl for 14 days, and Methiocarb and Prpoxur for 7 days. All of these pesticides were with detection limit of 3mugm(-3). It is suited for environmental monitoring. The airborne carbamates in different areas of the manufactory were measured.

  12. Stationary phase modulation in liquid chromatography through the serial coupling of columns: A review.

    PubMed

    Alvarez-Segura, T; Torres-Lapasió, J R; Ortiz-Bolsico, C; García-Alvarez-Coque, M C

    2016-06-01

    Liquid chromatography with single columns often does not succeed in the analysis of complex samples, in terms of resolution and analysis time. A relatively simple solution to enhance chromatographic resolution is the modulation of the stationary phase through the serial coupling of columns. This can be implemented with any type of column using compatible elution conditions and conventional instruments. This review describes the key features of column coupling and published procedures, where two or more columns were coupled in series to solve separation problems. In all reports, the authors could not resolve their samples with single columns, whereas significant enhancement in chromatographic performance was obtained when the columns were combined. Particularly interesting is the reduction in the analysis time in the isocratic mode, which alleviates the "general elution problem" of liquid chromatography, and may represent a stimulus for the proposal of new procedures, especially in combination with mass spectrometric, electrochemical and refractometric detection. Developments proposed to make the serial coupling of columns useful in routine and research laboratories are outlined, including optimisation strategies that facilitate the selection of the appropriate column combination and elution conditions (solvent content, flow rate or temperature) in both isocratic and gradient modes. The availability of zero dead volume couplers, able to connect standard columns, and the commercialisation of short columns with multiple lengths, have expanded the possibilities of success.

  13. Herbal medicine analysis by liquid chromatography/time-of-flight mass spectrometry.

    PubMed

    Zhou, Jian-Liang; Qi, Lian-Wen; Li, Ping

    2009-10-30

    The fact that the effects of herbal medicines (HMs) are brought about by their chemical constituents has created a critical demand for powerful analytical tools performing the chemical analysis to assure their efficacy, safety and quality. Liquid chromatography coupled to mass spectrometry (LC-MS) is an excellent technique to analyze multi-components in complex herbal matrices. Due to its inherent characteristics of accurate mass measurements and high resolution, time-of-flight (TOF) MS is well-suited to this field, especially for qualitative applications. The purpose of this article is to provide an overview on the potential of TOF, including the hybrid quadrupole- and ion trap-TOF (QTOF and IT-TOF), hyphenated to LC for chemical analysis in HMs or HM-treated biological samples. The peculiarities of LC-(Q/IT)TOF-MS for the analysis of HMs are discussed first, including applied stationary phase, mobile-phase selection, accurate mass measurements, fragmentation and selectivity. The final section is devoted to describing the applicability of LC-(Q/IT)TOF-MS to routine analysis of multi-components, including target and non-target (unknown) compounds, in herbal samples, emphasizing both the advantages and limitations of this approach for qualitative and quantitative purposes. The potential and future trends of fast high-performance liquid chromatography (HPLC) (e.g. rapid resolution LC and ultra-performance LC) coupled to (Q)TOF-MS for chemical analysis of HMs are highlighted.

  14. Determination of ochratoxin A in wine by immunoaffinity cleanup and liquid chromatography tandem mass spectrometry.

    PubMed

    Noba, Shigekuni; Omote, Masayuki; Kitagawa, Yasushi; Mochizuki, Naoki

    2008-05-01

    A simple and accurate method has been developed for determining ochratoxin A (OTA), using an immunoaffinity column for cleanup and liquid chromatography-tandem mass spectrometry for identification and quantification. Wine samples were diluted with a solution containing polyethylene glycol 8000 and sodium hydrogen carbonate, filtered through a glass microfiber filter, and cleaned up on an immunoaffinity column. OTA was then eluted with methanol-acetic acid (98:2) and analyzed by liquid chromatography-tandem mass spectrometry. The average recoveries of OTA from red and white wines were 95 and 96.7% (spiked OTA level was 0.05 ng/ml) and repeatabilities (relative standard deviation) were 3.8 and 2.4%, respectively. The detection limit was 0.0003 ng/ml based on the signal-to-noise ratio in wine of 3:1. Analysis of 74 samples of domestic and imported wines showed OTA levels ranging from < 0.0003 to 0.82 ng/ml, with an incidence of contamination of 92.1% for red wines, and < 0.0003 to 0.51 ng/ml, with an incidence of contamination of 77.8% for white wines. These detection rates were higher than those rates of past reports of OTA contamination in wine, due to the high sensitivity of this method. However, all samples analyzed in this study complied with European Union regulations. It is concluded that this method is a useful tool for the quality assurance of wine. PMID:18522043

  15. Flumethrin residue levels in honey from apiaries of China by high-performance liquid chromatography.

    PubMed

    Yu, L S; Liu, F; Wu, H; Tan, H R; Ruan, X C; Chen, Y; Chao, Z

    2015-01-01

    A method for detection of flumethrin residue in honey by high-performance liquid chromatography was established. After n-hexane-dichloromethane (4:6, vol/vol) extraction, the honey samples were concentrated by rotary evaporation, purified by an Oasis HLB solid-phase extraction column, and detected using a UV detector at 267 nm. The interference of the matrix was greatly reduced by optimizing pretreatment conditions; thus, the minimal detection limit of cyhalothrin was 0.005 mg/kg, the average recovery was 80.8 to 96.8%, and the coefficient of variation was 0.6 to 1.5%. The precision and reproducibility of this method was suitable and applicable for detecting flumethrin residue in honey. With this method, 135 honey samples from seven locations in the People's Republic of China were tested; 77 samples tested positive for flumethrin residue, resulting in a detection rate of 75.3%. Samples from the Guangdong province had the highest flumethrin residue level (0.122 mg/kg) of the locations tested. On the basis of analytical validation, the high-performance liquid chromatography has been shown to be a promising alternative for the analysis of flumethrin residue in honey samples.

  16. Analysis of aristolochic acids, aristololactams and their analogues using liquid chromatography tandem mass spectrometry.

    PubMed

    Yu, Jie; Ma, Chao-Mei; Wang, Xuan; Shang, Ming-Ying; Hattori, Masao; Xu, Feng; Jing, Yu; Dong, Shi-Wen; Xu, Yu-Qiong; Zhang, Cui-Ying; Cai, Shao-Qing

    2016-08-01

    More than 80 aristolochic acids (AAs) and aristololactams (ALs) have been found in plants of the Aristolochiaceae family, but relatively few have been fully studied. The present study aimed at developing and validating a liquid chromatography tandem mass spectrometry (LC/MS(n)) for the analysis of these compounds. We characterized the fragmentation behaviors of 31 AAs, ALs, and their analogues via high performance liquid chromatography coupled with electrospray ionization mass spectrometry. We summarized their fragmentation rules and used these rules to identify the constituents contained in Aristolochia contorta, Ar. debilis, Ar. manshurensis, Ar. fangchi, Ar. cinnabarina, and Ar. mollissima. The AAs and ALs showed very different MS behaviors. In MS(1) of AAs, the characteristic pseudomolecular ions were [M + NH4](+), [M + H](+), and [M + H - H2O](+). However, only [M + H](+) was found in the MS(1) of ALs, which was simpler than that of AAs. Distinct MS(n)fragmentation patterns were found for AAs and ALs, showing the same skeleton among the different substituent groups. The distribution of the 31 constituents in the 6 species of Aristolochia genus was reported for the first time. 25 Analogues of AAs and ALs were detected in this genus. A hierarchical schemes and a calculating formula of the molecular formula of these nitrophenanthrene carboxylic acids and their lactams were proposed. In conclusion, this method could be applied to identification of similar unknown constituents in other plants. PMID:27608953

  17. Detection of various freshwater cyanobacterial toxins using ultra-performance liquid chromatography tandem mass spectrometry.

    PubMed

    Oehrle, Stuart A; Southwell, Ben; Westrick, Judy

    2010-05-01

    Several freshwater cyanobacteria species have the capability to produce toxic compounds, frequently referred to as cyanotoxins. The most prevalent of these cyanotoxins is microcystin LR. Recognizing the potential health risk, France, Italy, Poland, Australia, Canada, and Brazil have set either standards or guidelines for the amount of microcystin LR permissible in drinking water based on the World Health Organization guideline of one microg/L of microcystin LR. Recently, the United States Environmental Protection Agency has begun to evaluate the occurrence and health effects of cyanotoxins and their susceptibility to water treatment under the Safe Drinking Water Act through the Contaminant Candidate List (CCL). A recent update of the Contaminant Candidate List focuses research and data collection on the cyanotoxins microcystin LR, anatoxin-a, and cylindrospermopsin. Liquid Chromatography/Tandem-Mass Spectrometry (LC/MS/MS) is a powerful tool for the analysis of various analytes in a wide variety of matrices because of its sensitivity and selectivity. The use of smaller column media (sub 2 microm particles) was investigated to both improve the speed, sensitivity and resolution, and to quantify the CCL cyanotoxins, in a single analysis, using Ultra-Performance Liquid Chromatography (UPLC) combined with tandem mass spectrometry. Natural waters and spiked samples were analyzed to show proof-of-performance. The presented method was able to clearly resolve each of the cyanotoxins in less than eight minutes with specificity and high spike recoveries. PMID:19878689

  18. Ethyl glucuronide determination in meconium and hair by hydrophilic interaction liquid chromatography-tandem mass spectrometry.

    PubMed

    Tarcomnicu, Isabela; van Nuijs, Alexander L N; Aerts, Katrien; De Doncker, Mireille; Covaci, Adrian; Neels, Hugo

    2010-03-20

    Ethyl glucuronide (EtG) detection in non-conventional matrices, such as hair and meconium, can provide useful information on alcohol abuse over a long time frame, for example during pregnancy or after a withdrawal treatment. This study reports on the development, validation and application of a new hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for the analysis of EtG in meconium and hair. For each matrix, the sample preparation and the chromatographic separation were thoroughly optimised. Additionally, experiments with reversed-phase liquid chromatography were also performed in the development stages. Analyses were carried out using a Phenomenex Luna HILIC column (150 mm x 3 mm, 5 microm) and a mobile phase composed by ammonium acetate 2mM and acetonitrile, in gradient. Different SPE cartridges (Oasis MAX, Oasis WAX, aminopropyl silica) and solvents were tested in order to obtain the highest recoveries and cleanest extracts. Optimal results were obtained for meconium with aminopropyl cartridges, while for hair an incubation of 16 h with 2 mL of water and acetonitrile (50/50, v/v) provided good results. The analytical method was validated for both matrices (meconium and hair) by assessing linearity, precision, accuracy, recovery and limit of quantification. The calibration curve concentrations ranged from 50 to 1200 pg/mg for meconium and from 20 to 1000 pg/mg for hair. Real meconium and hair samples were analyzed and results were consistent with literature. PMID:20061101

  19. Determination of bromate in drinking water by ultraperformance liquid chromatography-tandem mass spectrometry.

    PubMed

    Alsohaimi, Ibrahim Hotan; Alothman, Zeid Abdullah; Khan, Mohammad Rizwan; Abdalla, Mohammad Abulhassan; Busquets, Rosa; Alomary, Ahmad Khodran

    2012-10-01

    Bromate is a byproduct formed as a result of disinfection of bromide-containing source water with ozone or hypochlorite. The International Agency for Research on Cancer has recognized bromate as a possible human carcinogen, thus it is essential to determine in drinking water. Present work highlights a development of sensitive and fast analytical method for bromate determination in drinking water by using ultraperformance liquid chromatography-tandem mass spectrometry. The quality parameters of the developed method were established, obtaining very low limit of detection (0.01 ng/mL), repeatability and reproducibility have been found to be less than 3% in terms of relative standard deviation when analyzing a bromate standard at 0.05 μg/mL with 0.4 min analysis time. Developed method was applied for the analysis of metropolitan and bottled water from Saudi Arabia; 22 samples have been analyzed. Bromate was detected in the metropolitan water samples (from desalinization source) at concentrations ranging between 3.43 and 75.04 ng/mL and in the bottled water samples at concentrations ranging between 2.07 and 21.90 ng/mL. Moreover, in comparison to established analytical methods such as liquid chromatography-tandem mass spectrometry, the proposed method was found to be very sensitive, selective and rapid for the routine analysis of bromate at low level in drinking water. PMID:22815069

  20. New support for high-performance liquid chromatography based on silica coated with alumina particles.

    PubMed

    Silveira, José Leandro R; Dib, Samia R; Faria, Anizio M

    2014-01-01

    A new material based on silica coated with alumina nanoparticles was proposed for use as a chromatographic support for reversed-phase high-performance liquid chromatography. Alumina nanoparticles were synthesized by a sol-gel process in reversed micelles composed of sodium bis(2-ethylhexyl)sulfosuccinate, and the support material was formed by the self-assembly of alumina layers on silica spheres. Spectroscopic and (29)Si nuclear magnetic resonance results showed evidence of chemical bonds between the alumina nanoparticles and the silica spheres, while morphological characterizations showed that the aluminized silica maintained the morphological properties of silica desired for chromatographic purposes after alumina incorporation. Stability studies indicated that bare silica showed high dissolution (~83%), while the aluminized silica remained practically unchanged (99%) after passing one liter of the alkaline mobile phase, indicating high stability under alkaline conditions. The C18 bonded aluminized silica phase showed great potential for use in high-performance liquid chromatography to separate basic molecules in the reversed-phase mode.

  1. Muramic Acid Measurements for Bacterial Investigations in Marine Environments by High-Pressure Liquid Chromatography

    PubMed Central

    Mimura, Toru; Romano, Jean-Claude

    1985-01-01

    Muramic acid, a constituent of procaryotic cell walls, was assayed by high-pressure liquid chromatography in samples from several marine environments (water column, surface microlayer, and sediment) and a bacterial culture. It is used as a microbial biomass indicator. The method gave a good separation of muramic acid from interfering compounds with satisfactory reproducibility. A pseudomonad culture had a muramic acid content of 4.7 × 10−10 to 5.3 × 10−10 μg per cell during growth. In natural water samples, highly significant relationships were found between muramic acid concentrations and bacterial numbers for populations of 108 to 1011 cells per liter. The muramic acid content in natural marine water decreased from 5.3 × 10−10 to 1.6 × 10−10 μg per cell with increasing depth. In coastal sediments exposed to sewage pollution, concentrations of muramic acid, ATP, organic carbon, and total amino acids displayed a parallel decrease with increasing distance from the sewage outlet. Advantages of muramic acid measurement by high-pressure liquid chromatography are its high sensitivity and reduction of preparation steps, allowing a short time analysis. PMID:16346848

  2. A simple and simultaneous identification method for aloe, catechu and gambir by high performance liquid chromatography.

    PubMed

    Zhao, Yan; Kim, Young Ho; Lee, Wonjae; Lee, Young Keun; Kim, Kyung Tae; Kang, Jong Seong

    2016-01-01

    An effective and rapid method was developed for the simultaneous identification of aloe, catechu and gambir by high performance liquid chromatography-diode array detector (HPLC-DAD). Identification of three maker compounds presented in three medicinal materials was performed on high performance liquid chromatography-mass spectrometry (HPLC-MS). Under the optimal HPLC chromatographic conditions, sixty-two samples were processed on an Optimapak C18 column using a solvent system of acetonitrile (from 10% to 35%) and 0.1% phosphoric acid solution (from 90% to 65%) at a total flow rate of 1.0 mL/min and detected at 270 nm. All calibration curves exhibited good linear relationship (r(2)>0.9992). The relative standard deviation values of intra-day and inter-day precision were less than 1% and 2%, respectively. The recoveries of three analytes ranged from 99.48 to 100.97% with low RSDs (<2%). For the first time, this study demonstrates that the processed aloe, catechu and gambir are sold in local material markets in China and Korea without their correct identification. It indicates the existent of high potential medicinal risk by misuse of three medicinal materials. The developed HPLC method can be applied to prevent unexpected biological activity due to misapplication of medicinal materials. PMID:26342878

  3. High-throughput, Highly Sensitive Analyses of Bacterial Morphogenesis Using Ultra Performance Liquid Chromatography.

    PubMed

    Desmarais, Samantha M; Tropini, Carolina; Miguel, Amanda; Cava, Felipe; Monds, Russell D; de Pedro, Miguel A; Huang, Kerwyn Casey

    2015-12-25

    The bacterial cell wall is a network of glycan strands cross-linked by short peptides (peptidoglycan); it is responsible for the mechanical integrity of the cell and shape determination. Liquid chromatography can be used to measure the abundance of the muropeptide subunits composing the cell wall. Characteristics such as the degree of cross-linking and average glycan strand length are known to vary across species. However, a systematic comparison among strains of a given species has yet to be undertaken, making it difficult to assess the origins of variability in peptidoglycan composition. We present a protocol for muropeptide analysis using ultra performance liquid chromatography (UPLC) and demonstrate that UPLC achieves resolution comparable with that of HPLC while requiring orders of magnitude less injection volume and a fraction of the elution time. We also developed a software platform to automate the identification and quantification of chromatographic peaks, which we demonstrate has improved accuracy relative to other software. This combined experimental and computational methodology revealed that peptidoglycan composition was approximately maintained across strains from three Gram-negative species despite taxonomical and morphological differences. Peptidoglycan composition and density were maintained after we systematically altered cell size in Escherichia coli using the antibiotic A22, indicating that cell shape is largely decoupled from the biochemistry of peptidoglycan synthesis. High-throughput, sensitive UPLC combined with our automated software for chromatographic analysis will accelerate the discovery of peptidoglycan composition and the molecular mechanisms of cell wall structure determination.

  4. Determination of plastic additives in packaging by liquid chromatography coupled to high resolution mass spectrometry.

    PubMed

    Moreta, Cristina; Tena, María-Teresa

    2015-10-01

    A simple and sensitive analytical method for the determination of several plastic additives in multilayer packaging based on solid-liquid extraction (SLE) and ultra-high performance liquid chromatography (UHPLC) coupled to variable wavelength (VWD) and time of flight mass spectrometry (TOF-MS) detectors is presented. The proposed method allows the simultaneous determination of fourteen additives belonging to different families such as antioxidants, slip agents and light stabilizers, as well as two oxidation products in only 9min. The developed method was validated in terms of linearity, matrix effect error, detection and quantification limits, repeatability and intermediate precision. The instrumental method showed satisfactory repeatability and intermediate precision at concentrations closed to LOQ with RSDs less than 7 and 20%, respectively, and LODs until 5000 times more sensitive than other GC-FID and HPLC-VWD methods previously reported. Also, focused ultrasound solid-liquid extraction (FUSLE) was optimized and evaluated to extract plastic additives from packaging. Extraction results obtained by FUSLE and SLE were compared to those obtained by pressurized liquid extraction (PLE). All extraction methods showed excellent extraction efficiency for slip agents, however quantitative recovery of all analytes was achieved only by SLE with just 5ml of hexane for 10h. Finally, the selected method was applied to the analysis of packaging samples where erucamide, Irgafos 168, oxidized Irgafos 168, Irganox 1076 and Irganox 1010 were detected and quantified. PMID:26319625

  5. Determination of plastic additives in packaging by liquid chromatography coupled to high resolution mass spectrometry.

    PubMed

    Moreta, Cristina; Tena, María-Teresa

    2015-10-01

    A simple and sensitive analytical method for the determination of several plastic additives in multilayer packaging based on solid-liquid extraction (SLE) and ultra-high performance liquid chromatography (UHPLC) coupled to variable wavelength (VWD) and time of flight mass spectrometry (TOF-MS) detectors is presented. The proposed method allows the simultaneous determination of fourteen additives belonging to different families such as antioxidants, slip agents and light stabilizers, as well as two oxidation products in only 9min. The developed method was validated in terms of linearity, matrix effect error, detection and quantification limits, repeatability and intermediate precision. The instrumental method showed satisfactory repeatability and intermediate precision at concentrations closed to LOQ with RSDs less than 7 and 20%, respectively, and LODs until 5000 times more sensitive than other GC-FID and HPLC-VWD methods previously reported. Also, focused ultrasound solid-liquid extraction (FUSLE) was optimized and evaluated to extract plastic additives from packaging. Extraction results obtained by FUSLE and SLE were compared to those obtained by pressurized liquid extraction (PLE). All extraction methods showed excellent extraction efficiency for slip agents, however quantitative recovery of all analytes was achieved only by SLE with just 5ml of hexane for 10h. Finally, the selected method was applied to the analysis of packaging samples where erucamide, Irgafos 168, oxidized Irgafos 168, Irganox 1076 and Irganox 1010 were detected and quantified.

  6. [Simultaneous determination of pantothenic acid and D-panthenol in cosmetics by high performance liquid chromatography].

    PubMed

    Mao, Xiqin; Hu, Xia; Pan, Wei

    2010-11-01

    A high performance liquid chromatographic method (HPLC) and sample pretreatment method were developed for the simultaneous determination of pantothenic acid (vitamin B5) and D-panthenol (provitamin B5) in cosmetics with different matrices (including of creams, lotions, aqueous cosmetics, oily cosmetics, wax-based cosmetics, nail polish etc). A liquid-liquid extraction system composed of water and water-immiscible solvent was used to preliminarily separate the target components from other oil-soluble components and surfactants in cosmetics, then macromolecular water-soluble matrices in cosmetics were removed by coprecipitation with potassium ferrocyanide-zinc acetate precipitating agent, and then under acid condition, pantothenic acid and D-panthenol were enriched on a C18 solid-phase extraction sorbent. After the removal of other water-soluble impurities, target components were eluted by 40% methanol and then separated and quantitatively analyzed by high performance liquid chromatography with external standard method. Good linear relationship was achieved in the concentration range of 0.1-10 microg/g for pantothenic acid and D-panthenol. The linear correlation coefficients were separately 0.998 9 and 0.999 6. The average recoveries of the target components in cosmetics were more than 90%. Limit of detection of the method was 30 microg/g and the limit of quantification was 100 microg/g. This method can be used to simultaneously determine pantothenic acid and D-panthenol in cosmetics. The results are accurate and reliable. PMID:21381423

  7. Determination of N-nitrosamines by automated dispersive liquid-liquid microextraction integrated with gas chromatography and mass spectrometry.

    PubMed

    Amayreh, Mousa; Chanbasha, Basheer; Alhooshani, Khalid; Mu'azu, Nuhu Dalhat; Lee, Hian Kee

    2015-05-01

    An automated dispersive liquid-liquid microextraction integrated with gas chromatography and mass spectrometric procedure was developed for the determination of three N-nitrosamines (N-nitroso-di-n-propylamine, N-nitrosopiperidine, and N-nitroso di-n-butylamine) in water samples. Response surface methodology was employed to optimize relevant extraction parameters including extraction time, dispersive solvent volume, water sample pH, sodium chloride concentration, and agitation (stirring) speed. The optimal dispersive liquid-liquid microextraction conditions were 28 min of extraction time, 33 μL of methanol as dispersive solvent, 722 rotations per minute of agitation speed, 23% w/v sodium chloride concentration, and pH of 10.5. Under these conditions, good linearity for the analytes in the range from 0.1 to 100 μg/L with coefficients of determination (r(2) ) from 0.988 to 0.998 were obtained. The limits of detection based on a signal-to-noise ratio of 3 were between 5.7 and 124 ng/L with corresponding relative standard deviations from 3.4 to 5.9% (n = 4). The relative recoveries of N-nitroso-di-n-propylamine, N-nitrosopiperidine, and N-nitroso di-n-butylamine from spiked groundwater and tap water samples at concentrations of 2 μg/L of each analyte (mean ± standard deviation, n = 3) were (93.9 ± 8.7), (90.6 ± 10.7), and (103.7 ± 8.0)%, respectively. The method was applied to determine the N-nitrosamines in water samples of different complexities, such as tap water, and groundwater, before and after treatment, in a local water treatment plant.

  8. Performance Evaluation of three Liquid Chromatography Mass Spectrometry Methods for Broad Spectrum Drug Screening

    PubMed Central

    Lynch, Kara L.; Breaud, Autumn R.; Vandenberghe, Hilde; Wu, Alan H. B.; Clarke, William

    2010-01-01

    BACKGROUND Liquid chromatography-mass spectrometry (LC-MS) and tandem LC-MS (LC-MS/MS) are increasingly used in toxicology laboratories as a complementary method to gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-ultraviolet detection (LC-UV) for comprehensive drug screening (CDS). This study was designed to characterize the sensitivity and specificity of three LC-MS(/MS) vendor-supplied methods for targeted CDS and identify the current limitations associated with the use of these technologies. METHODS Five methods for broad spectrum CDS, including LC-UV (REMEDi), full scan GC-MS, LC-MS (ZQ™-Mass Detector with MassLynx™-software), LC-QTRAP-MS/MS (3200-QTRAP® with Cliquid®-software) and LC-LIT-MS/MS (LXQ™ Linear Ion Trap with ToxID™-software) were evaluated based on their ability to detect drugs in 48 patient urine samples. RESULTS The tandem MS methods identified 15% more drugs than the single stage MS or LC-UV methods. Use of two broad spectrum screening methods identified more drugs than any single system alone. False negatives and false positives generated by the LC-MS(/MS) software programs were identified upon manual review of the raw data. CONCLUSIONS The LC-MS/MS methods detected a broader menu of drugs; however, it is essential to establish manual data review criteria for all LC-MS(/MS) drug screening methods. Use of an EI-GC-MS and ESI-LC-MS/MS combination for targeted CDS may be optimal due to the complementary nature of the chromatographic and ionization techniques. PMID:20540936

  9. Determination of multiple phytohormones in fruits by high-performance liquid chromatography with fluorescence detection using dispersive liquid-liquid microextraction followed by precolumn fluorescent labeling.

    PubMed

    Li, Guoliang; Lu, Shuaimin; Wu, Hongliang; Chen, Guang; Liu, Shucheng; Kong, Xiaojian; Kong, Weiheng; You, Jinmao

    2015-01-01

    Plant hormone determination in food matrices has attracted more and more attention because of their potential risks to human health. However, analytical methods for the analysis of multiple plant hormones remain poorly investigated. In the present study, a convenient, selective, and ultrasensitive high-performance liquid chromatography method for the simultaneous determination of multiple classes of plant hormones has been developed successfully using dispersive liquid-liquid microextraction followed by precolumn fluorescent labeling. Eight plant hormones in fruits including jasmonic acid, 12-oxo-phytodienoic acid, indole-3-acetic acid, 3-indolybutyric acid, 3-indolepropionic acid, gibberellin A3 , 1-naphthylacetic acid, and 2-naphthaleneacetic acid were analyzed by this method. The conditions employed for dispersive liquid-liquid microextraction were optimized systematically. The linearity for all plant hormones was found to be >0.9993 (R(2) values). This method offered low detection limits of 0.19-0.44 ng/mL (at a signal-to-noise ratio of 3), and method accuracies were in the range of 92.32-103.10%. The proposed method was applied to determine plant hormones in five kinds of food samples, and this method can achieve a short analysis time, low threshold levels of detection, and a high specificity for the analysis of targeted plant hormones present at trace level concentrations in complex matrices.

  10. Determination of Aconitum Alkaloids in Dietary Supplements and Raw Botanical Materials by Liquid Chromatography/UV Detection with Confirmation by Liquid Chromatography/Tandem Mass Spectrometry: Collaborative Study

    PubMed Central

    Wong, Siu-Kay

    2010-01-01

    An interlaboratory study was conducted to evaluate a method for the determination of 3 Aconitum alkaloids, viz., aconitine, mesaconitine, and hypaconitine, in raw botanical material and dietary supplements. The alkaloids were extracted with diethyl ether in the presence of ammonia. After cleanup by solid-phase extraction to remove matrix interferences, the alkaloids were determined by reversed-phase liquid chromatography (LC)/UV detection at 235 nm with confirmation by LC/tandem mass spectrometry (MS/MS). A total of 14 blind duplicates were successfully analyzed by 12 collaborators. For repeatability, the relative standard deviation (RSDr) values ranged from 1.9 to 16.7%, and for reproducibility, the RSDR values ranged from 6.5 to 33%. The HorRat values were all <2 with only one exception at 2.3. All collaborating laboratories had calibration curves with correlation coefficients of >0.998. In addition, 6 collaborators performed the confirmation and were able to verify the identities of the alkaloids by using LC/MS/MS. PMID:19382567

  11. Separation and characterisation of beta2-microglobulin folding conformers by ion-exchange liquid chromatography and ion-exchange liquid chromatography-mass spectrometry.

    PubMed

    Bertoletti, Laura; Regazzoni, Luca; Aldini, Giancarlo; Colombo, Raffaella; Abballe, Franco; Caccialanza, Gabriele; De Lorenzi, Ersilia

    2013-04-10

    In this work we present for the first time the use of ion-exchange liquid chromatography to separate the native form and a partially structured intermediate of the folding of the amyloidogenic protein beta2-microglobulin. Using a strong anion-exchange column that accounts for the differences in charge exposure of the two conformers, a LC-UV method is initially optimised in terms of mobile phase pH, composition and temperature. The preferred mobile phase conditions that afford useful information were found to be 35 mM ammonium formate, pH 7.4 at 25°C. The dynamic equilibrium of the two species is demonstrated upon increasing the concentration of acetonitrile in the protein sample. Then, the chromatographic method is transferred to MS detection and the respective charge state distributions of the separated conformers are identified. The LC-MS results demonstrate that one of the conformers is partially unfolded, compared with the native and more compact species. The correspondence with previous results obtained in free solution by capillary electrophoresis suggest that strong ion exchange LC-MS does not alter beta2-microglobulin conformation and maintains the dynamic equilibrium already observed between the native protein and its folding intermediate. PMID:23522119

  12. High-performance liquid chromatography separation of unsaturated organic compounds by a monolithic silica column embedded with silver nanoparticles.

    PubMed

    Zhu, Yang; Morisato, Kei; Hasegawa, George; Moitra, Nirmalya; Kiyomura, Tsutomu; Kurata, Hiroki; Kanamori, Kazuyoshi; Nakanishi, Kazuki

    2015-08-01

    The optimization of a porous structure to ensure good separation performances is always a significant issue in high-performance liquid chromatography column design. Recently we reported the homogeneous embedment of Ag nanoparticles in periodic mesoporous silica monolith and the application of such Ag nanoparticles embedded silica monolith for the high-performance liquid chromatography separation of polyaromatic hydrocarbons. However, the separation performance remains to be improved and the retention mechanism as compared with the Ag ion high-performance liquid chromatography technique still needs to be clarified. In this research, Ag nanoparticles were introduced into a macro/mesoporous silica monolith with optimized pore parameters for high-performance liquid chromatography separations. Baseline separation of benzene, naphthalene, anthracene, and pyrene was achieved with the theoretical plate number for analyte naphthalene as 36,000 m(-1). Its separation function was further extended to cis/trans isomers of aromatic compounds where cis/trans stilbenes were chosen as a benchmark. Good separation of cis/trans-stilbene with separation factor as 7 and theoretical plate number as 76,000 m(-1) for cis-stilbene was obtained. The trans isomer, however, is retained more strongly, which contradicts the long- established retention rule of Ag ion chromatography. Such behavior of Ag nanoparticles embedded in a silica column can be attributed to the differences in the molecular geometric configuration of cis/trans stilbenes.

  13. Argentation high performance liquid chromatography on-line coupled to gas chromatography for the analysis of monounsaturated polyolefin oligomers in packaging materials and foods.

    PubMed

    Lommatzsch, Martin; Biedermann, Maurus; Simat, Thomas J; Grob, Koni

    2015-07-10

    Multidimensional chromatography based on two-dimensional high performance liquid chromatography on-line coupled to gas chromatography (on-line HPLC-HPLC-GC) enables the separate analysis of saturated, monounsaturated and aromatic hydrocarbons in packaging materials like polyolefins or paperboard and their migrates into foods. Since normal-phase HPLC on silica gel did not preseparate saturated from monounsaturated hydrocarbons, a separation step on a normal-phase HPLC column treated in the laboratory with an optimized amount of silver nitrate was added. The preparation of this HPLC column and the instrumental set-up are described, followed by examples showing the potential of the method. In a preliminary investigation of 11 polyolefin granulates for food contact up to 40% monounsaturated hydrocarbons among the oligomers C16-35 were determined.

  14. Analysis of captan, folpet, and captafol in apples by dispersive liquid-liquid microextraction combined with gas chromatography.

    PubMed

    Zang, Xiaohuan; Wang, Juntao; Wang, Ou; Wang, Mingzhao; Ma, Jingjun; Xi, Guohong; Wang, Zhi

    2008-10-01

    A novel method was developed for the determination of captan, folpet, and captafol in apples by dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-electron capture detection (GC-ECD). Some experimental parameters that influence the extraction efficiency, such as the type and volume of the disperser solvents and extraction solvents, extraction time, and addition of salt, were studied and optimized to obtain the best extraction results. Under the optimum conditions, high enrichment factors for the compounds were achieved ranging from 824 to 912. The recoveries of fungicides in apples at spiking levels of 20.0 microg kg(-1) and 70.0 microg kg(-1) were 93.0-109.5% and 95.4-107.7%, respectively. The relative standard deviations (RSDs) for the apple samples at 30.0 microg kg(-1) of each fungicide were in the range from 3.8 to 4.9%. The limits of detection were between 3.0 and 8.0 microg kg(-1). The linearity of the method ranged from 10 to 100 microg kg(-1) for the three fungicides, with correlation coefficients (r (2)) varying from 0.9982 to 0.9997. The obtained results show that the DLLME combined with GC-ECD can satisfy the requirements for the determination of fungicides in apple samples.

  15. Automated high performance liquid chromatography and liquid scintillation counting determination of pesticide mixture octanol/water partition rates

    SciTech Connect

    Moody, R.P.; Carroll, J.M.; Kresta, A.M.

    1987-12-01

    Two novel methods are reported for measuring octanol/water partition rates of pesticides. A liquid scintillation counting (LSC) method was developed for automated monitoring of /sup 14/C-labeled pesticides partitioning in biphasic water/octanol cocktail systems with limited success. A high performance liquid chromatography (HPLC) method was developed for automated partition rate monitoring of several constituents in a pesticide mixture, simultaneously. The mean log Kow +/- SD determined from triplicate experimental runs were for: 2,4-D-DMA (2,4-dichlorophenoxyacetic acid dimethylamine), 0.65 +/- .17; Deet (N,N-diethyl-m-toluamide), 2.02 +/- .01; Guthion (O,O-dimethyl-S-(4-oxo-1,2,3-benzotriazin-3(4H)-ylmethyl) phosphorodithioate), 2.43 +/- .03; Methyl-Parathion (O,O-dimethyl-O-(p-nitrophenyl) phosphorothioate), 2.68 +/- .05; and Fenitrothion (O,O-dimethyl O-(4-nitro-m-tolyl) phosphorothioate), 3.16 +/- .03. A strong positive linear correlation (r = .9979) was obtained between log Kow and log k' (log Kow = 2.35 (log k') + 0.63). The advantages that this automated procedure has in comparison with the standard manual shake-flask procedure are discussed.

  16. Estrogens determination in wastewater samples by automatic in-syringe dispersive liquid-liquid microextraction prior silylation and gas chromatography.

    PubMed

    González, Alba; Avivar, Jessica; Cerdà, Víctor

    2015-09-25

    A new procedure for the extraction, preconcentration and simultaneous determination of the estrogens most used in contraception pharmaceuticals (estrone, 17β-estradiol, estriol, and 17α-ethynylestradiol), cataloged as Contaminants of Emergent Concern by the Environmental Protection Agency of the United States (US-EPA), is proposed. The developed system performs an in-syringe magnetic stirring-assisted dispersive liquid-liquid microextraction (in-syringe-MSA-DLLME) prior derivatization and gas chromatography (GC-MS). Different extraction (carbon tetrachloride, ethyl acetate, chloroform and trichloroethylene) and disperser solvents (acetone, acetonitrile and methanol) were tested. Chloroform and acetone were chosen as extraction and disperser solvent, respectively, as they provided the best extraction efficiency. Then, a multivariate optimization of the extraction conditions was carried out. Derivatization conditions were also studied to ensure the conversion of the estrogens to their respective trimethylsilyl derivatives. Low LODs and LOQs were achieved, i.e. between 11 and 82ngL(-1), and 37 and 272ngL(-1), respectively. Good values for intra and inter-day precision were obtained (RSDs≤7.06% and RSD≤7.11%, respectively). The method was successfully applied to wastewater samples.

  17. Dual dispersive liquid-liquid microextraction for determination of phenylpropenes in oils by gas chromatography-mass spectrometry.

    PubMed

    Tsai, Chia-Ju; Li, Jih-Heng; Feng, Chia-Hsien

    2015-09-01

    A novel, simple and quick sample preparation method was developed and used for pre-concentration and extraction of six phenylpropenes, including anethole, estragole, eugenol, methyl eugenol, safrole and myristicin, from oil samples by dual dispersive liquid-liquid microextraction. Gas chromatography-mass spectrometry was used for determination and separation of compounds. Several experimental parameters affecting extraction efficiency were evaluated and optimized, including forward-extractant type and volume, surfactant type and concentration, water volume, and back-extractant type and volume. For all analytes (10-1000ng/mL), the limits of detection (S/N≧3) ranged from 1.0 to 3.0ng/mL; the limits of quantification (S/N≧10) ranged from 2.5 to 10.0ng/mL; and enrichment factors ranged from 3.2 to 37.1 times. Within-run and between-run relative standard deviations (n=6) were less than 2.61% and less than 4.33%, respectively. Linearity was excellent with determination coefficients (r(2)) above 0.9977. The experiments showed that the proposed method is a simple, effective, and environmentally friendly method of analyzing phenylpropenes in oil samples.

  18. High-performance liquid chromatography using pressurized liquid extraction for the determination of seven tetracyclines in egg, fish and shrimp.

    PubMed

    Liu, Yu; Yang, Hailan; Yang, Sheng; Hu, Qiwei; Cheng, Hongbo; Liu, Huiyu; Qiu, Yinsheng

    2013-02-15

    A simple and especially rapid method, pressurized liquid extraction, has been developed and applied to the quantitative determination of oxytetracycline, tetracycline, chlortetracycline, minocycline, methacycline, demeclocycline and doxycycline in egg, fish and shrimp. The procedure consisted of a trichloracetic acid/methanol extraction conducted at elevated temperature (60°C) and pressure (65bar), without further clean-up, the extraction solution was concentrated and finally for high-performance liquid chromatography analysis. The limits of detection were 5.0-10.0μg/kg and the limits of quantification were 10.0-15.0μg/kg for tetracyclines in egg, fish and shrimp using UV detection. The analytical limits CCα and CCβ were also calculated. The recoveries of tetracyclines spiked at levels of 15-300μg/kg, averaged 75.6-103.5% with the relative standard deviation values less than 11%. The optimized procedure has been successfully applied to real samples in our laboratories. It demonstrated that the new method was robust and useful for monitoring and quantification of 7 tetracycline residues in food of animal origin. PMID:23353938

  19. Defluorinated Sparfloxacin as a New Photoproduct Identified by Liquid Chromatography Coupled with UV Detection and Tandem Mass Spectrometry

    PubMed Central

    Engler, Michael; Rüsing, Guido; Sörgel, Fritz; Holzgrabe, Ulrike

    1998-01-01

    Photodegradation of sparfloxacin was observed by means of high-pressure liquid chromatography with UV detection and liquid chromatography coupled with UV detection and tandem mass spectrometry (LC-MS/MS). Three products were detected. Comparison with an independently synthesized derivative of sparfloxacin revealed the structure of one product which is believed to be 8-desfluorosparfloxacin. The second product is likely to be formed by the splitting off of a fluorine and a cyclopropyl ring. Thus, photodefluorination of quinolone antibacterial agents is found and proved for the first time by LC-MS/MS. PMID:9593143

  20. Optimization of the Hewlett-Packard particle-beam liquid chromatography-mass spectrometry interface by statistical experimental design.

    PubMed

    Huang, S K; Garza, N R

    1995-06-01

    Optimization of both sensitivity and ionization softness for the Hewlett-Packard particle-beam liquid chromatography-mass spectrometry interface has been achieved by using a statistical experimental design with response surface modeling. Conditions for both optimized sensitivity and ionization softness were found to occur at 55-lb/in.(2) nebulizer flow, 35°C desolvation chamber temperature with approximately 45% organic modifier in the presence of 0.02-F ammonium acetate and a liquid chromatography flow rate of 0.2 mL/min.

  1. Single-step electrodeposition of CIS thin films with the complexing agent triethanolamine

    NASA Astrophysics Data System (ADS)

    Chiu, Yu-Shuen; Hsieh, Mu-Tao; Chang, Chih-Min; Chen, Chun-Shuo; Whang, Thou-Jen

    2014-04-01

    Some difficulties have long been encountered by single-step electrodeposition such as the optimization of electrolyte composition, deposition potentials, deposition time, and pH values. The approach of introducing ternary components into single-step electrodeposition is rather challenging especially due to the different values of the equilibrium potential for each constituent. Complexing agents play an important role in single-step electrodeposition of CuInSe2 (CIS), since the equilibrium potential of every constituent can be brought closer to each other when complexing agents are employed. In this work, single-step electrodeposition of CIS was enhanced by adding triethanolamine (TEA) into deposition bath, the CIS thin films were improved consequently in the form of polycrystalline cauliflower structures through the examination of SEM images and XRD patterns. The optimum composition of the solution for single-step electrodeposition of CIS is found to be 5 mM CuCl2, 22 mM InCl3, and 22 mM SeO2 at pH 1.5 with 0.1 M TEA. The structures, compositions, and morphologies of as-deposited and of annealed films were investigated.

  2. Neuere Chromatographie

    NASA Astrophysics Data System (ADS)

    Hostettmann, K.

    1983-04-01

    Besides high-performance liquid chromatography (HPLC) which is now a well-established and currently used technique, several emerging methods for the isolation and separation of natural products are receiving considerable attention. Centrifugal thin-layer chromatography is a very rapid technique, but limited in resolution. Of special interest are the recently developed support-free liquid-liquid chromatography methods such as droplet counter-current chromatography (DCCC) and rotation locular counter-current chromatography (RLCC). This latter method was applied to the separation of the enantiomers of (±)-norephedrine.

  3. Ultra-high-pressure liquid chromatography-tandem mass spectrometry method for the determination of alkylphenols in soil.

    PubMed

    Wang, Jing; Pan, Hefang; Liu, Zhengzheng; Ge, Fei

    2009-03-20

    A novel method has been developed for the determination of alkylphenols in soil by ultra-high-pressure liquid chromatography employing small particle sizes, combined with tandem mass spectrometry. Soil samples were extracted with pressurized liquid extraction (PLE) and then cleaned with solid-phase extraction (SPE). The extracts were separated on C18 column (1.7 microm, 50 mm x 2.1mm) with a gradient elution and a mobile phase consisting of water and acetonitrile, and then detected by an electrospray ionization tandem mass spectrometry in negative ion mode with multiple reaction monitoring (MRM). Compared with traditional liquid chromatography, it took ultra-high-pressure liquid chromatography much less time to analyze alkylphenols. Additionally, the ultra-high-pressure liquid chromatography/tandem mass spectrometry method produces satisfactory reliability, sensitivity, and accuracy. The average recoveries of the three target analytes were 74.0-103.4%, with the RSD<15%. The calibration curves for alkylphenols were linear within the range of 0.01-0.4 microg/ml, with the correlation coefficients greater than 0.99. When 10 g soil sample was used for analysis, the limits of quantification (LOQs) of the three alkylphenols were all 1.0 microg/kg.

  4. Community air monitoring for pesticides-part 2: multiresidue determination of pesticides in air by gas chromatography, gas chromatography-mass spectrometry, and liquid chromatography-mass spectrometry.

    PubMed

    Hengel, Matt; Lee, P

    2014-03-01

    Two multiresidue methods were developed to determine pesticides in air collected in California. Pesticides were trapped using XAD-4 resin and extracted with ethyl acetate. Based on an analytical method from the University of California Davis Trace Analytical Laboratory, pesticides were detected by analyzing the extract by gas chromatography-mass spectrometry (GC-MS) to determine chlorothalonil, chlorthal-dimethyl, cycloate, dicloran, dicofol, EPTC, ethalfluralin, iprodione, mefenoxam, metolachlor, PCNB, permethrin, pronamide, simazine, trifluralin, and vinclozolin. A GC with a flame photometric detector was used to determine chlorpyrifos, chlorpyrifos oxon, diazinon, diazinon oxon, dimethoate, dimethoate oxon, fonophos, fonophos oxon, malathion, malathion oxon, naled, and oxydemeton. Trapping efficiencies ranged from 78 to 92 % for low level (0.5 μg) and 37-104 % for high level (50 and 100 μg) recoveries. Little to no degradation of compounds occurred over 31 days; recoveries ranged from 78 to 113 %. In the California Department of Food and Agriculture (CDFA) method, pesticides were detected by analyzing the extract by GC-MS to determine chlorothalonil, chlorpyrifos, cypermethrin, dichlorvos, dicofol, endosulfan 1, endosulfan sulfate, oxyfluorfen, permethrin, propargite, and trifluralin. A liquid chromatograph coupled to a MS was used to determine azinphos-methyl, chloropyrifos oxon, DEF, diazinon, diazinon oxon, dimethoate, dimethoate oxon, diuron, EPTC, malathion, malathion oxon, metolachlor, molinate, norflurazon, oryzalin, phosmet, propanil, simazine and thiobencarb. Trapping efficiencies for compounds determined by the CDFA method ranged from 10 to 113, 22 to 114, and 56 to 132 % for 10, 5, and 2 μg spikes, respectively. Storage tests yielded 70-170 % recovery for up to 28 days. These multiresidue methods represent flexible, sensitive, accurate, and cost-effective ways to determine residues of various pesticides in ambient air. PMID:24370860

  5. Simultaneous Determination of Eight Bioactive Compounds in Dianthus superbus by High-performance Liquid Chromatography

    PubMed Central

    Yun, Bo-Ra; Yang, Hye Jin; Weon, Jin Bae; Lee, Jiwoo; Eom, Min Rye; Ma, Choong Je

    2016-01-01

    Background: Dianthus superbus, one of traditional herbal medicine, is widely used to treat urethritis, carbuncles and carcinoma. Objective: A simultaneous determination method was established for controlling the quality of D. superbus using the eight compounds, (E)-methyl-4-hydroxy-4-(8a-methyl-3-oxodecahydronaphthalen-4a-yl) (1), diosmetin-7-O(2'',6''-di-O-α-L-rhamnopyranosyl)-β-D-glucopyranoside (2), vanillic acid (3), 4-hydroxyphenyl acetic acid (4), 4-methoxyphenyl acetic acid (5), (E)-4-methoxycinnamic acid (6), 3-methoxy-4-hydroxyphenylethanol (7), and methyl hydroferulate (8) isolated from D. superbus. Materials and Methods: This analysis method was developed using high performance liquid chromatography coupled with diode array detector with a Shishedo C18 column at a column temperature of 3°C. The mobile phase was composed of 0.1% trifluoroacetic acid in water and acetonitrile. The flow rate was 1 ml/min and detection wavelength was set at 205 nm and 280 nm. Validation was performed in order to demonstrate selectivity, accuracy and precision of the method. Results: The calibration curves showed good linearity (R2 > 0.99). The limits of detection and limits of quantification were within the ranges 0.0159–0.6205 μg/ml and 0.3210–1.8802 μg/ml, respectively. Moreover, the relative standard deviations of intra- and inter-day precision were both <2.98%. The overall recoveries were in the range of 96.23–109.87%. Quantitative analysis of eight compounds in 12 D. superbus samples (D-1–D-12) from various regions were analyzed and compared by developed method. Conclusion: As a result, this established method was accurate and sensitive for the quality evaluation of eight compounds isolated from D. superbus and may provide a new basis for quality control of D. superbus. SUMMARY A simultaneous determination method of eight compounds in Dianthus superbus was established by high performance liquid chromatography-diode array detectorDeveloped analysis method is

  6. Characterization of enhanced-fluidity liquid hydrophilic interaction chromatography for the separation of nucleosides and nucleotides.

    PubMed

    Philibert, Gwenaëlle S; Olesik, Susan V

    2011-11-11

    Hydrophilic interaction chromatography (HILIC) is a liquid chromatographic separation mechanism commonly used for polar biological molecules. The use of enhanced-fluidity liquid chromatography (EFLC) with mixtures of methanol/water/carbon dioxide is compared to acetonitrile/water mobile phases for the separation of nucleosides and nucleotides under HILIC conditions. Enhanced-fluidity liquid chromatography involves using common mobile phases with the addition of substantial proportions of a dissolved gas which provides greater mobile phase diffusivity and lower viscosity. The impact of varying several experimental parameters, including temperature, addition of base, salt, and CO₂ was studied to provide optimized HILIC separations. Each of these parameters plays a key role in the retention of the analytes, which demonstrates the complexity of the retention mechanism in HILIC. The tailing of phosphorylated compounds was overcome with the use of phosphate salts and the addition of a strong base; efficiency and peak asymmetry were compared with the addition of either triethylamine (TEA), 1,4-diazabicyclo [2.2.2] octane (DABCO) or 1,5-diazabicyclo [4.3.0] non-5-ene (DBN). DBN and DABCO both led to increased efficiency and lower peak asymmetry; DBN provided the best results. Sodium chloride and carbon dioxide were added to enhance the selectivity between the analytes, giving a successful isocratic separation of nucleosides and nucleotides within 8 min. The retention mechanism involved in EFL-HILIC was explored by varying the temperature and the mole fraction of CO₂. These studies showed that partitioning was the dominant mechanism. The thermodynamics study confirmed that the solvent strength is maintained in EFLC and that a change in entropy was mainly responsible for the improved selectivity. The selectivity using methanol/water/carbon dioxide varied greatly compared to that obtained with acetonitrile/water. Finally while this study highlights the optimization of EFL

  7. High-performance liquid chromatography method for analyzing the antiretroviral agent efavirenz in human plasma.

    PubMed

    Villani, P; Pregnolato, M; Banfo, S; Rettani, M; Burroni, D; Seminari, E; Maserati, R; Regazzi, M B

    1999-06-01

    Efavirenz (EFV, DMP-266) is a new antiretroviral agent belonging to the class of nonnucleoside reverse transcriptase inhibitors. It has recently been approved by the Food and Drug Administration in management of human immunodeficiency virus (HIV). Preliminary pharmacokinetic studies on EFV in healthy volunteers show that the drug may influence the metabolism of protease inhibitors. For the determination of EFV in human plasma, a validated and specific reverse-phase high-performance liquid chromatography (HPLC) method, with UV detection, was developed. We used 100 microL plasma sample for a liquid-liquid extraction with diethyl ether after basification. The mobile phase was a mixture of acetonitrile and water, pumped at a flow rate of 1.2 mL/min. Ultraviolet detection was carried out at a wavelength of 247 nm. Retention times for EFV and internal standard (IS) were 5.3 and 4.5 minutes, respectively, and there was no chromatographic interference from other commonly administered drugs. The limit of detection was 100 ng/mL. The described assay is a rapid and accurate method for measurement of EFV in plasma: the easy preparation and small sample size makes this assay highly suitable for pharmacokinetic studies and routine clinical analysis in patients with HIV. In addition, the reproducibility of the method is only moderately increased by including IS, so analyzing without IS may be an alternative.

  8. Quantification of testosterone undecanoate in human hair by liquid chromatography-tandem mass spectrometry.

    PubMed

    Pozo, Oscar J; Deventer, Koen; Van Eenoo, Peter; Rubens, Robert; Delbeke, Frans T

    2009-08-01

    Testosterone undecanoate (T-C11) can be used by athletes in order to improve performance. After oral intake, T-C11 is rapidly metabolized, hampering discrimination between exogenous and endogenous testosterone. A possible alternative is to detect the intact ester in hair. A method based on liquid chromatography-tandem mass spectrometry was developed for the determination of T-C11 in hair. The sample procedure consisted of digestion of 200 mg of pulverized hair with tris(2-carboxyethyl)phosphine hydrochloride and liquid-liquid extraction with n-pentane. Several parameters such as the mobile phase, the ionization source and the washing step were optimized. The method was validated at different spiked levels obtaining satisfactory values for accuracy (between 92 and 102%) with relative standard deviations lower than 7% and a limit of detection of 0.2 ng/g. The applicability of the method was checked by the analysis of three samples from patients using T-C11. A peak for the analyte was detected in all samples with concentrations between 0.4 and 8.4 ng/g. PMID:19353724

  9. Confirmation and quantification of clenbuterol in horse urine using liquid chromatography tandem mass spectrometry triple quadrupole.

    PubMed

    Bishop, Jennifer; Heffron, Brendan; Taddei, Lisa; Benoit, Marc; Hurt, Laura; Costello, Sara; Gross, Melissa; Negrusz, Adam

    2015-03-01

    Clenbuterol (CLE) is used in horses as a bronchodilator and for its anabolic steroid-like effects. CLE is a Class 3 drug according to current Association of Racing Commissioners International (ARCI) Uniform Classification Guidelines. The Racing Medication and Testing Consortium recommended a urine CLE threshold of 140 pg/mL after careful scientific review of the results of studies describing the disposition of CLE in the horse and this threshold was adopted by the ARCI. Enzyme-linked immunosorbent assay was previously used to screen samples for CLE in Illinois, but could not detect such low concentrations in urine. Thus, a liquid-liquid extraction of CLE from urine followed by quantification by liquid chromatography-tandem mass spectrometry was developed and validated. Method validation included testing stability, ion suppression and enhancement, precision, accuracy and uncertainty. Intra-, interday and total precision and accuracy were calculated for each control and found to be within the ±15% acceptance range. The Guide to the Expression of Uncertainty in Measurement approach was used to calculate uncertainty, which was 11% at the 95% confidence level. In the past 5 years, only 15 samples were reported as positive for CLE in Illinois. This new method was used in a pilot program to screen and confirm samples received from thoroughbred and harness horses.

  10. [Determination of 12 sulfonamides in cosmetics by ultra performance liquid chromatography].

    PubMed

    Zheng, Hehui; Wang, Ping; Li, Jie

    2007-03-01

    A method for the determination of 12 sulfonamides (SAs) (sulfanilamide, sulfamonomethoxine, sulfacetamide, sulfamethoxazole, sulfadiazine, sulfisoxazole, sulfathiazole, sulfadi-methoxine, sulfamerazine, sulfaquinoxaline, sulfamethazine, sulfanitran) in cosmetics was developed by ultra performance liquid chromatography with photodiode array detector (UPLC-PDA). The chromatographic column used was Acquity UPLC BEHC C18 (50 mm x 2. 1 mm, 1. 7 microm) and the mobile phase was acetonitrile-0. 1% formic acid aqueous solution. A gradient elution program was utilized for the separation and determination. After liquid-liquid extraction, SAs were separated and detected by UPLC-PDA. The qualification analysis was done by using retention time and spectrum, and the quantification was based on the detection wavelength of 268 nm. The limits of qualification (S/N = 3) and quantification (S/N = 10) for 12 SAs were 1 microg/g and 2 -3 microg/g, respectively. The correlation coefficient of linear calibration curve was over 0. 999 7 within the SAs concentration range of 1 - 25 mg/L (except sulfanitran 0. 5 - 12. 5 mg/L). At the spiked levels of 40 and 8 microg (except sulfanitran 20 and 4 microg), the average recoveries for 12 SAs were 86. 8% - 98. 1% and 80. 1% - 96. 9%, respectively. Relative standard deviations were less than 10%. Routine tests show that the method is simple, fast, and has a good separation efficiency. It can be routinely used for the determination of these SAs in cosmetics. PMID:17580695

  11. Determination of ethyl glucuronide in human hair by hydrophilic interaction liquid chromatography-tandem mass spectrometry.

    PubMed

    Yaldiz, Fadile; Daglioglu, Nebile; Hilal, Ahmet; Keten, Alper; Gülmen, Mete Korkut

    2013-10-01

    Ethyl glucuronide (EtG) is a direct metabolite of ethanol and has been utilized as a marker for alcohol intake. This study presents development, validation and application of a new hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method for the analysis of EtG in human hair samples. The linearity was assessed in the range of 5-2000 pg/mg hair, with a correlation coefficient of >0.99. The method was selective and sensitive, with a limit of detection (LOD) and limit of quantitation (LOQ) of 0.05 pg/mg and 0.18 pg/mg in hair, respectively. Differently from the extraction procedures in the literature, a fast and simple liquid-liquid method was used and highest recoveries and cleanest extracts were obtained. The method was successfully applied to 30 human hair samples which were taken from those who state they consume alcohol. EtG concentrations in the hair samples of alcohol users participated in this study, ranged between 1.34 and 82.73 pg/mg. From the concentration of EtG in hair strands 20 of the 30 subjects can be considered regular moderate drinkers. PMID:24112322

  12. [Determination of thermodynamic parameters for ionic liquid 1-hexyl-3-methylimidazolium trifluoromethansulfonate by inverse gas chromatography].

    PubMed

    Deng, Lishuang; Wang, Qiang; Zhang, Zhengfang; Tang, Jun

    2014-02-01

    The thermodynamic parameters of ionic liquid 1-hexyl-3-methylimidazolium trifluoromethansulfonate ([HMIM] OTF) were investigated by inverse gas chromatography in the temperature range of 343.15-373.15 K. Eighteen probe solvents were used to calculate the molar enthalpy of sorption, molar enthalpy of mixing at infinite dilution, molar enthalpy of vaporization and the mass fraction activity coefficients. Furthermore, Flory-Huggins interaction parameters, the solubility parameter of the ionic liquid were calculated to judge the interactions between [HMIM] OTF and the 18 probes solvents. The results showed that among the selected solvents, n-C6-9, tetrahydrofuran, diethyl ether, cyclohexane and benzene are the poor solvents for [HMIM] OTF, while dichloromethane, acetone, chloroform, ethyl acetate, carbon tetrachloride, methyl acetate, toluene and methanol are the favorite solvents for [HMIM] OTF. In addition, the solubility parameter of [HMIM] OTF at room temperature (298.15 K), which was obtained by linear extrapolation method, was 20.74 (J/cm3)0.5. This study could be used as a reference to the application and research of the ionic liquids. PMID:24822452

  13. Analysis of phytosterols in extra-virgin olive oil by nano-liquid chromatography.

    PubMed

    Rocco, Anna; Fanali, Salvatore

    2009-10-23

    In this work the applicability of nano-liquid chromatography (nano-LC) was evaluated for the determination of phytosterols in extra-virgin olive oil samples. These compounds represent a minor part of lipids in vegetable oils, but their quantification can be useful to establish oil origin and to reveal intentional adulterations. The analysis of five main sterols, specifically brassicasterol, stigmasterol, campesterol, cholesterol and beta-sitosterol, was performed in a laboratory-assembled nano-LC system coupled with a UV detector. The separation of all compounds was obtained in about 20 min, employing a capillary column packed with a C18-RP (sub-2 microm particles) stationary phase for 15 cm. Methanol only was used as mobile phase. The simple method developed and optimized was validated in terms of repeatability, linearity, limit of detection and limit of quantification (0.78 and 1.56 microg/mL, respectively) achieving good results. After this, it was applied to the determination of phytosterols in extra-virgin olive oil samples. Isolation of phytosterols was obtained by solid-phase extraction, after saponification and liquid-liquid extraction of the unsaponified fraction with diethyl ether. Recovery tests were performed and values between 90% and 103%, with RSDs within 5%, were obtained. Moreover the nano-LC system was coupled with a mass spectrometer for an accurate identification of phytosterols. PMID:19386314

  14. Highly efficient peptide separations in proteomics Part 1. Unidimensional high performance liquid chromatography.

    PubMed

    Sandra, Koen; Moshir, Mahan; D'hondt, Filip; Verleysen, Katleen; Kas, Koen; Sandra, Pat

    2008-04-15

    Sample complexity and dynamic range constitute enormous challenges in proteome analysis. The back-end technology in typical proteomics platforms, namely mass spectrometry (MS), can only tolerate a certain complexity, has a limited dynamic range per spectrum and is very sensitive towards ion suppression. Therefore, component overlap has to be minimized for successful mass spectrometric analysis and subsequent protein identification and quantification. The present review describes the advances that have been made in liquid-based separation techniques with focus on the recent developments to boost the resolving power. The review is divided in two parts; the first part deals with unidimensional liquid chromatography and the second part with bi- and multidimensional liquid-based separation techniques. Part 1 mainly focuses on reversed-phase HPLC due to the fact that it is and will, in the near future, remain the technique of choice to be hyphenated with MS. The impact of increasing the column length, decreasing the particle diameter, replacing the traditional packed beds by monolithics, amongst others, is described. The review is complemented with data obtained in the laboratories of the authors. PMID:18006394

  15. LC/ESI-MS determination of proteins using conventional liquid chromatography and ultrasonically assisted electrospray.

    PubMed

    Banks, J F; Quinn, J P; Whitehouse, C M

    1994-11-01

    Conventional unassisted electrospray ionization mass spectrometry (ESI-MS) has severe limitations as a liquid chromatography (LC) interface due to the few solvent compositions which can be electrosprayed without some type of assistance. LC mobile phases which have high flow rates (over 5 microL/min), high conductivity, or high surface tension are normally unsuitable for use with ESI. For this reason, an ultrasonic nebulizer has been developed which mechanically creates the fine spray of droplets needed for ESI and thus functions independently of the solvent composition. This device also operates at considerably higher liquid flow rates, up to several hundred microliters per minute. To characterize the system's performance, the ultrasonic nebulizer frequency, source electrode potentials, and drying and focusing gas flow rates were studied and optimized. Also, droplet size measurements were taken using a phase-Doppler anemometer, which showed consistent nebulizer performance up to a liquid flow rate of several hundred microliters per minute. Finally, the ultrasonic nebulizer was used for an LC application involving the separation of proteins on 1.0 and 2.1 mm i.d. columns. LC flow rates of up to 200 microL/min of mobile phases containing 0.1% trifluoroacetic acid could be easily nebulized. With the 1.0 mm i.d. column, as little as 32 fmol of cytochrome c could be detected in selected ion monitoring mode.

  16. A Simple and Universal Gel Permeation Chromatography Technique for Precise Molecular Weight Characterization of Well-Defined Poly(ionic liquid)s

    SciTech Connect

    He, Hongkun; Zhong, Mingjiang; Adzima, Brian; Luebke, David; Nulwala, Hunaid; Matyjaszewski, Krzysztof

    2013-03-20

    Poly(ionic liquid)s (PILs) are an important class of technologically relevant materials. However, characterization of well-defined polyionic materials remains a challenge. Herein, we have developed a simple and versatile gel permeation chromatography (GPC) methodology for molecular weight (MW) characterization of PILs with a variety of anions. PILs with narrow MW distributions were synthesized via atom transfer radical polymerization, and the MWs obtained from GPC were further confirmed via nuclear magnetic resonance end group analysis.

  17. Self-assembled cyclodextrin-modified gold nanoparticles on silica beads as stationary phase for chiral liquid chromatography and hydrophilic interaction chromatography.

    PubMed

    Li, Yuanyuan; Wei, Manman; Chen, Tong; Zhu, Nan; Ma, Yulong

    2016-11-01

    A facile strategy based on self-assembly of Au nanoparticles (AuNPs) (60±10nm in size) on the surfaces of amino-functionalized porous silica spheres under mild conditions was proposed. The resulting material possessed a core-shell structure in which AuNPs were the shell and silica spheres were the core. Then, thiolated-β-cyclodextrin (SH-β-CD) was covalently attached onto the AuNPs as chiral selector for the enantioseparation. The resultant packing material was evaluated by high-performance liquid chromatography (HPLC). The separations of nine pairs of enantiomers were achieved by using the new chiral stationary phase (CSP) in the reversed-phase liquid chromatography (RPLC) mode, respectively. The results showed the new CSP have more sufficient interaction with the analytes due to the existence of AuNPs on silica surfaces, resulting in faster mass transfer rate, compared with β-CD modified silica column. The result shed light on potential usage of chemical modified NPs as chiral selector for enantioseparation based on HPLC. In addition, the new phase was also used in hydrophilic interaction liquid chromatography (HILIC) to separate polar compounds and highly hydrophilic compounds. PMID:27591589

  18. On-line hyphenation of centrifugal partition chromatography and high pressure liquid chromatography for the fractionation of flavonoids from Hippophaë rhamnoides L. berries.

    PubMed

    Michel, Thomas; Destandau, Emilie; Elfakir, Claire

    2011-09-01

    Centrifugal Partition Chromatography (CPC), a liquid-liquid preparative chromatography using two immiscible solvent systems, benefits from numerous advantages for the separation or purification of synthetic or natural products. This study presents the on-line hyphenation of CPC-Evaporative Light Scattering Detector (CPC-ELSD) with High Performance Liquid Chromatography-UV (HPLC-UV) for the fractionation of flavonols from a solvent-free microwave extract of sea buckthorn (Hippophaë rhamnoides L., Elaeagnaceae) berries. An Arizona G system was used for the fractionation of flavonoids by CPC and a fused core Halo C18 column allowed the on-line analyses of collected fractions by HPLC. The on-line CPC/HPLC procedure allowed the simultaneous fractionation step at preparative scale combined with the HPLC analyses which provide direct fingerprint of collected fractions. Thus the crude extract was simplified and immediate information on the composition of fractions could be obtained. Furthermore, this methodology reduced the time of post-fractionation steps and facilitated identification of main molecules by Mass Spectrometry (MS). Rutin, isorhamnetin-3-O-rutinoside, isorhamnetin-3-O-glucoside, quercetin-3-O-glucoside, isorhamnetin-rhamnoside, quercetin and isorhamnetin were identified. CPC-ELSD/HPLC-UV could be considered as a high-throughput technique for the guided fractionation of bioactive natural products from complex crude extracts.

  19. A dispersive liquid-liquid micellar microextraction for the determination of pharmaceutical compounds in wastewaters using ultra-high-performace liquid chromatography with DAD detection.

    PubMed

    Montesdeoca-Esponda, Sarah; Mahugo-Santana, Cristina; Sosa-Ferrera, Zoraida; Santana-Rodríguez, José Juan

    2015-03-01

    A dispersive liquid-liquid micellar microextraction (DLLMME) method coupled with ultra-high-performance liquid chromatography (UHPLC) using Diode Array Detector (DAD) detector was developed for the analysis of five pharmaceutical compounds of different nature in wastewaters. A micellar solution of a surfactant, polidocanol, as extraction solvent (100 μL) and chloroform as dispersive solvent (200 μL) were used to extract and preconcentrate the target analytes. Samples were heated above critical temperature and the cloudy solution was centrifuged. After removing the chloroform, the reduced volume of surfactant was then injected in the UHPLC system. In order to obtain high extraction efficiency, the parameters affecting the liquid-phase microextraction, such as time and temperature extraction, ionic strength and surfactant and organic solvent volume, were optimized using an experimental design. Under the optimized conditions, this procedure allows enrichment factors of up to 47-fold. The detection limit of the method ranged from 0.1 to 2.0 µg/L for the different pharmaceuticals. Relative standard deviations were <26% for all compounds. The procedure was applied to samples from final effluent collected from wastewater treatment plants in Las Palmas de Gran Canaria (Spain), and two compounds were measured at 67 and 113 µg/L in one of them. PMID:25056775

  20. Liquid chromatography-tandem mass spectrometric assay for the tyrosine kinase inhibitor afatinib in mouse plasma using salting-out liquid-liquid extraction.

    PubMed

    Sparidans, Rolf W; van Hoppe, Stephanie; Rood, Johannes J M; Schinkel, Alfred H; Schellens, Jan H M; Beijnen, Jos H

    2016-02-15

    A quantitative bioanalytical liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay for afatinib, an irreversible inhibitor of the ErbB (erythroblastic leukemia viral oncogene homolog) tyrosine kinase family, was developed and validated. Plasma samples were pre-treated using salting-out assisted liquid-liquid extraction (SALLE) with acetonitrile, magnesium chloride and a stable isotopically labeled internal standard. After dilution, the extract was directly injected into the reversed-phase liquid chromatographic system. The eluate was transferred into the electrospray interface with positive ionization and compounds were detected in the selected reaction monitoring mode of a triple quadrupole mass spectrometer. The assay was completely validated for plasma in a 0.5-500ng/ml calibration range with r(2)=0.995±0.002 (n=6) using linear regression with the inverse square of the concentration as the weighting factor for the calibration. Within-run precisions (n=18) were 2.7-11.7% and between-run (3 runs; n=18) precisions 3.0-14.5%. Accuracies were between 96-109% for the whole calibration range. The drug was sufficiently stable under all relevant analytical conditions. Finally, the assay was successfully applied to determine plasma drug levels and study pharmacokinetics after oral administration of afatinib to female FVB mice.

  1. Determination of chlorophenols in honey samples using in-situ ionic liquid-dispersive liquid-liquid microextraction as a pretreatment method followed by high-performance liquid chromatography.

    PubMed

    Fan, Chen; Li, Nai; Cao, Xueli

    2015-05-01

    In-situ ionic liquid-dispersive liquid-liquid microextraction (IL-DLLME) method was developed as a pretreatment method for the detection of six chlorophenols (CPs) in honey samples. The hydrophobic ionic liquid [C4MIM][NTf2], formed in-situ by the hydrophilic ionic liquid [C4MIM][BF4] and the ion exchange reagent LiNTf2 was used as the microextractant solvent of CPs from honey sample. Then the enriched analytes were back-extracted into 40 μL of 0.14 M NaOH solution and finally subjected to analysis by high-performance liquid chromatography. The method showed low limit of detection of CPs, 0.8-3.2 μg/L and high enrichment factor, 34-65 with the recoveries range from 91.60% to 114.33%. The method is simple, rapid, environmentally friendly and with high extraction efficiency.

  2. Improvement of mineral oil saturated and aromatic hydrocarbons determination in edible oil by liquid-liquid-gas chromatography with dual detection.

    PubMed

    Zoccali, Mariosimone; Barp, Laura; Beccaria, Marco; Sciarrone, Danilo; Purcaro, Giorgia; Mondello, Luigi

    2016-02-01

    Mineral oils, which are mainly composed of saturated hydrocarbons and aromatic hydrocarbons, are widespread food contaminants. Liquid chromatography coupled to gas chromatography with flame ionization detection represents the method of choice to determine these two families. However, despite the high selectivity of this technique, the presence of olefins (particularly squalene and its isomers) in some samples as in olive oils, does not allow the correct quantification of the mineral oil aromatic hydrocarbons fraction, requiring additional off-line tools to eliminate them. In the present research, a novel on-line liquid chromatography coupled to gas chromatography method is described for the determination of hydrocarbon contamination in edible oils. Two different liquid chromatography columns, namely a silica one (to retain the bulk of the matrix) and a silver-ion one (which better retains the olefins), were coupled in series to obtain the mineral oil aromatic hydrocarbons hump free of interfering peaks. Furthermore, the use of a simultaneous dual detection, flame ionization detector and triple quadrupole mass spectrometer allowed us not only to quantify the mineral oil contamination, but also to evaluate the presence of specific markers (i.e. hopanes) to confirm the petrogenic origin of the contamination.

  3. Improvement of mineral oil saturated and aromatic hydrocarbons determination in edible oil by liquid-liquid-gas chromatography with dual detection.

    PubMed

    Zoccali, Mariosimone; Barp, Laura; Beccaria, Marco; Sciarrone, Danilo; Purcaro, Giorgia; Mondello, Luigi

    2016-02-01

    Mineral oils, which are mainly composed of saturated hydrocarbons and aromatic hydrocarbons, are widespread food contaminants. Liquid chromatography coupled to gas chromatography with flame ionization detection represents the method of choice to determine these two families. However, despite the high selectivity of this technique, the presence of olefins (particularly squalene and its isomers) in some samples as in olive oils, does not allow the correct quantification of the mineral oil aromatic hydrocarbons fraction, requiring additional off-line tools to eliminate them. In the present research, a novel on-line liquid chromatography coupled to gas chromatography method is described for the determination of hydrocarbon contamination in edible oils. Two different liquid chromatography columns, namely a silica one (to retain the bulk of the matrix) and a silver-ion one (which better retains the olefins), were coupled in series to obtain the mineral oil aromatic hydrocarbons hump free of interfering peaks. Furthermore, the use of a simultaneous dual detection, flame ionization detector and triple quadrupole mass spectrometer allowed us not only to quantify the mineral oil contamination, but also to evaluate the presence of specific markers (i.e. hopanes) to confirm the petrogenic origin of the contamination. PMID:26614690

  4. Simple high-performance liquid chromatography method for formaldehyde determination in human tissue through derivatization with 2,4-dinitrophenylhydrazine.

    PubMed

    Yilmaz, Bilal; Asci, Ali; Kucukoglu, Kaan; Albayrak, Mevlut

    2016-08-01

    A simple high-performance liquid chromatography method has been developed for the determination of formaldehyde in human tissue. FA Formaldehyde was derivatized with 2,4-dinitrophenylhydrazine. It was extracted from human tissue with ethyl acetate by liquid-liquid extraction and analyzed by high-performance liquid chromatography. The calibration curve was linear in the concentration range of 5.0-200 μg/mL. Intra- and interday precision values for formaldehyde in tissue were <6.9%, and accuracy (relative error) was better than 6.5%. The extraction recoveries of formaldehyde from human tissue were between 88 and 98%. The limits of detection and quantification of formaldehyde were 1.5 and 5.0 μg/mL, respectively. Also, this assay was applied to liver samples taken from a biopsy material.

  5. Use of aluminum phosphate as the dehydration catalyst in single step dimethyl ether process

    DOEpatents

    Peng, Xiang-Dong; Parris, Gene E.; Toseland, Bernard A.; Battavio, Paula J.

    1998-01-01

    The present invention pertains to a process for the coproduction of methanol and dimethyl ether (DME) directly from a synthesis gas in a single step (hereafter, the "single step DME process"). In this process, the synthesis gas comprising hydrogen and carbon oxides is contacted with a dual catalyst system comprising a physical mixture of a methanol synthesis catalyst and a methanol dehydration catalyst. The present invention is an improvement to this process for providing an active and stable catalyst system. The improvement comprises the use of an aluminum phosphate based catalyst as the methanol dehydration catalyst. Due to its moderate acidity, such a catalyst avoids the coke formation and catalyst interaction problems associated with the conventional dual catalyst systems taught for the single step DME process.

  6. Single step, rapid identification of pathogenic microorganisms in a culture bottle.

    PubMed

    Chu, Yu W; Wang, Bo Y; Engebretson, David A; Carey, James R

    2013-10-21

    Efforts to treat bloodstream infections, which have a relatively high mortality rate, are delayed by the lengthy multi-step process required to identify the causative bacteria. Due to this delay, broad spectrum antibiotics are prescribed on a presumptive basis, leading to the rise of antibiotic resistant microorganisms. Here, as proof of principle, we describe a colourimetric sensor that rapidly identifies opportunistic pathogenic bacteria in a single step in TSB media. The device is composed of a reaction chamber and an array of chemoresponsive dyes deposited on a substrate in a prearranged pattern. This single step, disposable, automated system can detect and identify of eight strains of bacteria, starting with clinically relevant concentrations bacteria in twenty four hours in TSB media. Thus, this technology may be used to streamline the current blood culture process by combining detection and identification in a single step.

  7. Determination of Aristolochic Acid in Botanicals and Dietary Supplements by Liquid Chromatography with Ultraviolet Detection and by Liquid Chromatography/Mass Spectrometry: Single Laboratory Validation Confirmation

    PubMed Central

    Trujillo, William A.; Sorenson, Wendy R.; La Luzerne, Paul; Austad, John W.; Sullivan, Darryl

    2008-01-01

    The presence of aristolochic acid in some dietary supplements is a concern to regulators and consumers. A method has been developed, by initially using a reference method as a guide, during single laboratory validation (SLV) for the determination of aristolochic acid I, also known as aristolochic acid A, in botanical species and dietary supplements at concentrations of approximately 2 to 32 μg/g. Higher levels were determined by dilution to fit the standard curve. Through the SLV, the method was optimized for quantification by liquid Chromatography with ultraviolet detection (LC-UV) and LC/mass Spectrometry (MS) confirmation. The test samples were extracted with organic solvent and water, then injected on a reverse phase LC column. Quantification was achieved with linear regression using a laboratory automation system. The SLV study included systematically optimizing the LC-UV method with regard to test sample size, fine grinding of solids, and solvent extraction efficiency. These parameters were varied in increments (and in separate optimization studies), in order to ensure that each parameter was individually studied; the test results include corresponding tables of parameter variations. In addition, the chromatographic conditions were optimized with respect to injection volume and detection wavelength. Precision studies produced overall relative standard deviation values from 2.44 up to 8.26% for aristolochic acid I. Mean recoveries were between 100 and 103% at the 2 μg/g level, between 102 and 103% at the 10 μg/g level, and 104% at the 30 μg/g level. PMID:16915829

  8. Polar organic phase liquid chromatography with packed capillary columns using a vancomycin chiral stationary phase

    PubMed

    Svensson; Donnecke; Karlsson; Karlsson; Vessman

    2000-08-01

    Vancomycin immobilized on silica served as the chiral stationary phase (CSP) in this investigation with polar organic solvents as the mobile phase in liquid chromatography (LC). It was shown that trace amounts of water were beneficial for improving peak shape and efficiency. To regulate the retention and selectivity an acid and/or base were added to the mobile phase where an excess of acid was shown to be preferential for enantioseparation. An unusual increase in selectivity with increasing temperature was shown for the acidic drug, thalidomide. Additionally, nonlinear van't Hoff plots were obtained for metoprolol enantiomers that showed increased retention with increasing temperature. Metoprolol also showed unusual behavior in the polar organic phase when water was added to resemble reversed-phase chromatography, with minimum retention observed at high water or high methanol concentrations. In both instances a high degree of electrostatic interaction between metoprolol and vancomycin was concluded. Metoprolol and ten of its analogs were examined on this CSP to evaluate the enantiorecognition process. A comparison in enantioselectivity for a number of acidic and basic drugs using this CSP was also carried out using the polar organic phase, reversed phase, and normal phase LC which were all compared to the results obtained in supercritical fluid chromatography (SFC). Polar organic phase LC offered a better separation of basic molecules while reversed phase LC was preferred for the resolution of acids. SFC showed the broadest enantioselectivity overall and normal phase LC indicated similar properties, as expected, to SFC but with lower column efficiency. Copyright 2000 Wiley-Liss, Inc. PMID:10897097

  9. Measurement of oxidative stress parameters using liquid chromatography-tandem mass spectroscopy (LC-MS/MS)

    SciTech Connect

    Winnik, Witold M. Kitchin, Kirk T.

    2008-11-15

    There is increasingly intense scientific and clinical interest in oxidative stress and the many parameters used to quantify the degree of oxidative stress. However, there remain many analytical limitations to currently available assays for oxidative stress markers. Recent improvements in software, hardware, and instrumentation design have made liquid chromatography and tandem mass spectroscopy (LC-MS/MS) methods optimal choices for the determination of many oxidative stress markers. In particular, LC-MS/MS often provides the advantages of higher specificity, higher sensitivity, and the capacity to determine multiple analytes (e.g. 4-11 oxidative stress markers per LC run) when compared to other available methods, such as gas chromatography-MS, immunoassays, spectrophotometric or flourometric assays. LC-MS/MS methods are also compatible with cleanup and sample preparation methods including prior solid phase extraction or automated two dimensional LC/LC chromatography followed by MS/MS. LC-MS/MS provides three analytical filtering functions: (1) the LC column provides initial separation as each analyte elutes from the column. (2) The first MS dimension isolates ions of a particular mass-to-charge (m/z) ratio. (3) The selected precursor ion is fragmented into product ions that provide structural information about the precursor ion. Quantitation is achieved based on the abundances of the product ions. The sensitivity limits for LC-MS/MS usually lie within the range of fg-pg of analyte per LC on-column injection. In this article, the present capabilities of LC-MS/MS are briefly presented and some specific examples of the strengths of these LC-MS/MS assays are discussed. The selected examples include methods for isoprostanes, oxidized proteins and amino acids, and DNA biomarkers of oxidative stress.

  10. Measurement of oxidative stress parameters using liquid chromatography-tandem mass spectroscopy (LC-MS/MS).

    PubMed

    Winnik, Witold M; Kitchin, Kirk T

    2008-11-15

    There is increasingly intense scientific and clinical interest in oxidative stress and the many parameters used to quantify the degree of oxidative stress. However, there remain many analytical limitations to currently available assays for oxidative stress markers. Recent improvements in software, hardware, and instrumentation design have made liquid chromatography and tandem mass spectroscopy (LC-MS/MS) methods optimal choices for the determination of many oxidative stress markers. In particular, LC-MS/MS often provides the advantages of higher specificity, higher sensitivity, and the capacity to determine multiple analytes (e.g. 4-11 oxidative stress markers per LC run) when compared to other available methods, such as gas chromatography-MS, immunoassays, spectrophotometric or fluorometric assays. LC-MS/MS methods are also compatible with cleanup and sample preparation methods including prior solid phase extraction or automated two dimensional LC/LC chromatography followed by MS/MS. LC-MS/MS provides three analytical filtering functions: (1) the LC column provides initial separation as each analyte elutes from the column. (2) The first MS dimension isolates ions of a particular mass-to-charge (m/z) ratio. (3) The selected precursor ion is fragmented into product ions that provide structural information about the precursor ion. Quantitation is achieved based on the abundances of the product ions. The sensitivity limits for LC-MS/MS usually lie within the range of fg-pg of analyte per LC on-column injection. In this article, the present capabilities of LC-MS/MS are briefly presented and some specific examples of the strengths of these LC-MS/MS assays are discussed. The selected examples include methods for isoprostanes, oxidized proteins and amino acids, and DNA biomarkers of oxidative stress. PMID:18547599

  11. A data comparison between a traditional and the single-step β-galactosidase assay.

    PubMed

    Schaefer, Jorrit; Jovanovic, Goran; Kotta-Loizou, Ioly; Buck, Martin

    2016-09-01

    This article describes reproducibility of a single-step automated β-galactosidase, and the equivalence of its data to the traditional assay ("Experiments in Molecular Genetics" [1]). This was done via a pairwise comparison of both methods using strains with Miller Unit [MU] values ranging from 0 to over 2000. The data presented in this article is associated with the research article entitled "A single-step method for mid to high throughput β-galactosidase assays in Escherichia coli using a microplate reader" [2].

  12. Liquid chromatography/electrospray ionization mass spectrometric characterization of Harpagophytum in equine urine and plasma.

    PubMed

    Colas, Cyril; Garcia, Patrice; Popot, Marie-Agnès; Bonnaire, Yves; Bouchonnet, Stéphane

    2006-01-01

    A method has been developed for the analysis and characterization in equine urine and plasma of iridoid glycosides: harpagide, harpagoside and 8-para-coumaroyl harpagide, which are the main active principles of Harpagophytum, a plant with antiinflammatory properties. The method involves liquid chromatography coupled with positive electrospray ionization mass spectrometry. The addition of sodium or lithium chloride instead of formic acid in the eluting solvent has been studied in order to enhance the signal and to modify the ion's internal energy. Fragmentation pathways and associated patterns are proposed for each analyte. A comparison of three types of mass spectrometer: a 3D ion trap, a triple quadrupole and a linear ion trap, has been conducted. The 3D ion trap was selected for drug screening analysis whereas the linear ion trap was retained for identification and quantitation analysis. PMID:17044124

  13. Uncovering biologically significant lipid isomers with liquid chromatography, ion mobility spectrometry and mass spectrometry

    SciTech Connect

    Kyle, Jennifer E.; Zhang, Xing; Weitz, Karl K.; Monroe, Matthew E.; Ibrahim, Yehia M.; Moore, Ronald J.; Cha, Jeeyeon; Sun, Xiaofei; Lovelace, Erica S.; Wagoner, Jessica; Polyak, Stephen J.; Metz, Thomas O.; Dey, Sudhansu K.; Smith, Richard D.; Burnum-Johnson, Kristin E.; Baker, Erin S.

    2016-01-01

    Understanding how biological molecules are generated, metabolized and eliminated in living systems is important for interpreting processes such as immune response and disease pathology. While genomic and proteomic studies have provided vast amounts of information over the last several decades, interest in lipidomics has also grown due to improved analytical technologies revealing altered lipid metabolism in type 2 diabetes, cancer, and lipid storage disease. Liquid chromatography and mass spectrometry (LC-MS) measurements are currently the dominant approach for characterizing the lipidome by providing detailed information on the spatial and temporal composition of lipids. However, interpreting lipids’ biological roles is challenging due to the existence of numerous structural and stereoisomers (i.e. distinct acyl chain and double-bond positions), which are unresolvable using present LC-MS approaches. Here we show that combining structurally-based ion mobility spectrometry (IMS) with LC-MS measurements distinguishes lipid isomers and allows insight into biological and disease processes.

  14. Determination of metformin and its prodrugs in human and rat blood by hydrophilic interaction liquid chromatography.

    PubMed

    Huttunen, Kristiina M; Rautio, Jarkko; Leppänen, Jukka; Vepsäläinen, Jouko; Keski-Rahkonen, Pekka

    2009-10-15

    Simple and specific hydrophilic interaction liquid chromatography (HILIC) method with ultraviolet (UV) detection was developed for the simultaneous determination of highly water-soluble metformin and its more lipophilic prodrugs in human and rat blood samples. The sample preparation was accomplished by precipitating proteins with acetonitrile, which enabled the direct injection of supernatants to the HPLC. Chromatographic separation was performed on an analytical normal phase silica column using a mixture of 0.01 M ammonium acetate pH 5.0 and acetonitrile (40:60, v/v) as a mobile phase at flow rate of 1 ml/min and at the wavelength of 235 nm. The method was validated in terms of specificity, linearity, accuracy, precision, recovery, and analyte stability. The UV-HILIC method was suitable for detecting both metformin and one of its more lipophilic prodrugs simultaneously in human and rat blood samples.

  15. Cobalt determination in natural waters using cation-exchange liquid chromatography with luminol chemiluminescence detection

    SciTech Connect

    Boyle, E.A.; Handy, B.; van Geen, A.

    1987-06-01

    A method has been developed for the analysis of cobalt in natural waters by cation-exchange liquid chromatography using chemiluminescence detection. Cobalt can be determined directly in freshwater samples on 500-..mu..L samples with a detection limit of 20 pmol/kg; larger samples provide proportionately lower detection limits. Seawater samples can be analyzed on 100-mL samples following APDC solvent extraction; the detection limit of this method is 5 pmol/kg. The precision of the method is +/- 5%. The method should also be applicable to the analysis of V, Cu, and Fe in natural waters. Equipment is low in cost and transportable and can be used in the field.

  16. Separation of amino acids by high performance liquid chromatography based on calixarene-bonded stationary phases.

    PubMed

    Zadmard, Reza; Tabar-Heydar, Kourosh; Imani, Maryam

    2015-01-01

    In this work, we present a new method for synthesis of silica gel stationary phases based on calix[4]arene derivatives. In order to achieve it, 25,27-dipropoxy-26,28-bis-[3-propyloxydimethylsililoxy]calix[4]arene has been synthesized in six steps and immobilized on silica via chlorotrimethylsilane. Stationary phases were characterized by elemental analysis, infrared spectroscopy and thermal analysis and used for the separation of amino acid derivatives by high performance liquid chromatography. The effect of isocratic and gradient elution, pH and column temperature on retention and selectivity of the Fmoc-protected amino acids were studied. The retention mechanism was also discussed. The results indicated that the stationary phase behaves like a reverse phase packing. Size exclusion, electron-π, π-π and hydrophobic interactions seem to be involved in the separation process.

  17. [Determination of three anabolic steroids by liquid chromatography-mass spectrometry].

    PubMed

    Qin, Yang; Liu, Xin; Wang, Zhanliang; Wu, Moutian

    2008-07-01

    A high performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI/MS) was developed to screen and determine trenbolone, tetrahydrogestrinone and gestrinone in human urine. The urine sample was enzymatically hydrolyzed with beta-glucuronidase, then extracted with methyl tert-butyl ether. Chromatographic separation was performed on a Zorbax SB-C18 column (150 mm x 2.1 mm, 5 microm) with ammonium formate buffer (pH 3.5) and acetonitrile as mobile phase. Using positive electrospray ionization mode, the effect of different parameters from electrospray chamber was investigated. The limits of detection based on signal noise ratio of 3 were between 1 and 5 ng. The method can be applied in screening and confirmation of the anabolic steroids in doping control.

  18. [Simultaneous analysis of ten anabolic steroids in blood plasma using high performance liquid chromatography].

    PubMed

    Zhang, Lan; Chen, Jinfeng; Tong, Ping; Li, Tianlin

    2008-07-01

    A method for the simultaneous separation and determination of ten anabolic steroids in blood plasma using high performance liquid chromatography (HPLC) was established. An RP-C18 column was used as the analytical column, and the mixture of acetonitrile and water was used as the mobile phase with gradient elution according to the characteristics of the analytes. The analytes were detected at the adjustable wavelengths ranging from 194 to 290 nm. Under the optimal conditions, ten compounds were separated within 10 min. The detection limits were in the range of 0.01 - 0.10 microg/mL. The spiked recoveries of standards in a rabbit plasma sample were from 70.3% to 120%. Methandriol was injected into the ear meridian of a rabbit, and then the anabolic steroid methandriol in the plasma was successfully detected with the established method. The results show that the method is feasible, rapid, simple and accurate.

  19. Determination of amphetamine and methamphetamine in umbilical cord using liquid chromatography-tandem mass spectrometry

    PubMed Central

    Jones, Joseph; Rios, Rosemarie; Jones, Mary; Lewis, Douglas; Plate, Charles

    2009-01-01

    The use of meconium as a drug-screening matrix for newborns has been the gold standard of care for the past two decades. A recent study using matched pairs of meconium and umbilical cord demonstrated a high degree of agreement. The use of liquid chromatography-tandem mass spectrometry as a means to confirm amphetamines presumptive positive umbilical cord specimens for amphetamine and methamphetamine is described here for the first time. The limit of detection for both compounds was 0.2 ng/g. The limit of quantitation for both compounds was 0.6 ng/g. The assay was linear for both compounds up to 100 ng/g. PMID:19783234

  20. Enzyme immunoassay and liquid chromatography-fluorescence detection for amikacin in raw milk.

    PubMed

    Chen, Yiqiang; Chen, Qian; He, Lidong; Shang, Bingru; Zhang, Liying

    2012-11-15

    An enzyme immunoassay and a liquid chromatography (LC) method for amikacin (AMK) in raw milk were developed in this study. Anti-AMK antibody was prepared by immunizing rabbits with AMK-BSA conjugate. The developed immunoassay exhibited an IC(50) value of 1.30 ng/mL and the spiked recoveries at 25-1000 ng/mL ranged from 69.8% to 93.3% with coefficients of variation (CVs) of 8.5-17.6%. For LC analysis, AMK was derivatized with 9-fluorenylmethyl chloroformate, which was followed by C8 column separation and fluorescence detection. By trichloroacetic acid extraction and MCX column purification, the recoveries at spiked concentrations of 500-5000 ng/mL were 80.7-91.3% with CVs less than 6.3%. The two methods can be selectively used for rapidly screening or quantitatively determining AMK in raw milk. PMID:22868103