• Title/Summary/Keyword: Liquid chromatography assay

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Development of a Denaturing High-Performance Liquid Chromatography (DHPLC) Assay to Detect Parasite Infection in Grass Shrimp Palaemonetes pugio

  • Cho, Sang-Man
    • Fisheries and Aquatic Sciences
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    • v.15 no.2
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    • pp.107-115
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    • 2012
  • In developing a useful tool to detect parasitic dynamics in an estuarine ecosystem, a denaturing high-performance liquid chromatography (DHPLC) assay was optimized by cloning plasmid DNA from the grass shrimp Palaemonetes pugio, and its two parasites, the trematode Microphallus turgidus and bopyrid isopod Probopyrus pandalicola. The optimal separation condition was an oven temperature of $57.9^{\circ}C$ and 62-68% of buffer B gradient at a flow rate of 0.45 mL/min. A peptide nucleic acid blocking probe was designed to clamp the amplification of the host gene, which increased the amplification efficiency of genes with low copy numbers. Using the DHPLC assay with wild-type genomic, the assay could detect GC Gram positive bacteria and the bopyrid isopod (P. pandalicola). Therefore, the DHPLC assay is an effective tool for surveying parasitic dynamics in an estuarine ecosystem.

Antioxidant Activity Analysis of Useful Compounds from Artemisiae Annuae Herba Using On-line Screening HPLC-ABTS+ Assay (On-line Screening HPLC-ABTS+ assay를 이용한 청호로부터 유용성분의 항산화 활성 분석)

  • Lee, Kwang Jin;Ma, Jin Yeul
    • Journal of Applied Biological Chemistry
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    • v.57 no.4
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    • pp.301-305
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    • 2014
  • The Antioxidant activity screening identification of five kind compounds in Artemisiae annuae herba with the on-line screening high performance liquid chromatography (HPLC) $ABTS^+$ assay. The various experimental variables such as the extraction time (h) and extraction solvent composition (%) of dipping method were investigated efficiently extraction at the room temperature $25^{\circ}C$. The results, the highest yield of total extract amount (0.458 g, 15.250%) was obtained by dipping method with 100% water and extraction time to 3 h. And the on-line screening HPLC-$ABTS^+$ assay method was rapid and efficient to search for bioactivity from natural products.

Rapid Quantification of Topotecan in Biological Samples by Liquid Chromatography/Tandem Mass Spectrometry

  • Shin, Beom-Soo;Lee, Mann-Hyung;Yoo, Sun-Dong
    • Journal of Pharmaceutical Investigation
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    • v.39 no.5
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    • pp.367-372
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    • 2009
  • A rapid liquid chromatography/tandem mass spectrometry (LC/MS/MS) assay method was developed for the determination of topotecan levels in rat serum. The assay utilized a single liquid-liquid extraction with a mixture of ethy l acetate and acetonitrile (6:1 v/v) and isocratic elution. The multiple reaction monitoring was based on the transition of m/z 422.0$\rightarrow$376.5 for topotecan and 315.1$\rightarrow$226.6 for clomipramine (internal standard). The developed assay was validated to demonstrate the specificity, recovery, lower limit of quantification (LLOQ), accuracy and precision. The assay was linear over a concentration range from 0.5-100 ng/mL, with LLOQ being 0.5 ng/mL using a small volume of rat serum (0.1 mL). The mean intra- and inter-day assay accuracy was 87.7-111.0% and 97.8-108.3, respectively, and the mean intra- and interday precision was between 1.6-4.3% and 3.8-10.3, respectively. The developed assay was applied to a pharmacokinetic study after a bolus i.v. injection of topotecan in rats.

Microcystin Detection Characteristics of Fluorescence Immunochromatography and High Performance Liquid Chromatography

  • Pyo, Dong-Jin;Park, Geun-Young;Choi, Jong-Chon;Oh, Chang-Suk
    • Bulletin of the Korean Chemical Society
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    • v.26 no.2
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    • pp.268-272
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    • 2005
  • Different detection characteristics of fluorescence immunochromatography method and high performance liquid chromatography (HPLC) method for the analysis of cyanobacterial toxins were studied. In particular, low and high limits of detection, detection time and reproducibility and detectable microcystin species were compared when fluorescence immunochromatography method and high performance liquid chromatography method were applied for the detection of microcystin (MC), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa. A Fluorescence immunochromatography assay system has the unique advantages of short detection time and low detection limit, and high performance liquid chromatography detection method has the strong advantage of individual quantifications of several species of microcystins.

Evaluation of Anti-Oxidant from Natural Products (천연물로부터 유래한 천연 항산화제 규명)

  • Kwon, Jin-A;Yang, Yoon-Jung;Park, Jong-Hyuk;Kang, Se-Chan
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2011.10a
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    • pp.20-20
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    • 2011
  • In this study, we analyzed 80%MeOH extract of fruits of sorbaria sorbifolia var. stellipila MAX. to measure the total antioxidant capacity by oxygen radical absorbance capacity (ORAC) assay, individual flavonoid content by high-performance liquid chromatography (HPLC). n-Hexane ($1.02{\pm}0.036$), $CH_2Cl_2$ ($0.95{\pm}0.025$), EtOAc ($1.94{\pm}0.065$), n-BuOH ($1.98{\pm}0.054$), D.W. ($1.2{\pm}0.032$) fractions were examined antioxidative activity by ORAC assay. It was revealed that EtOAc($1.94{\pm}0.065$), n-BuOH($1.98{\pm}0.054$) fractions had significant antioxidative activity. The isolation and separation were facilitated using open column chromatography, while separation, purification and identification were accomplished by using high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR).

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Analysis of Vicamine Using High Performance Liquid Chromatography and Antioxidant Activity of Vincaminor Extract (High performance liquid chromatography를 이용한 빈카민 분석 및 빈카마이너의 항산화능 측정)

  • Jung, Jong-Hee;Back, Yu-Mi;Lee, Kwang-Geun
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.599-602
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    • 2008
  • Vincamine, one of the major indole alkaloids in vincaminor (Vinca minor L.) is commonly used for treating cerebrovascular diseases. The antioxidant activity of vincaminor extracts and vincamine were measured by 1.1-diphenyl-2-picrylhydrazyl (DPPH) and lipid malonaldehyde (MA) assay. Vincaminor leaves were pulverized and extracted with various solvents such as water, methanol, and ethanol. The antioxidant activities of the extracts varied in accordance with solvents and assays. In DPPH assay, the water extract showed the highest antioxidant activity. In lipid MA assay, However, the ethanol extract inhibited MA formation from cod liver oil by 82% at the level of 5,000 ${\mu}g/mL$. Vincamine in the extract was analyzed by high-performance liquid chromatogram and the concentration of vincamine was 0.419$\pm$0.005 ${\mu}g/mL$.

Bioactivity Analysis of Curcuminoids from Turmeric using On-line Screening HPLC-ABTS (On-line Screening HPLC-ABTS를 이용한 강황으로부터 Curcuminoids의 생물활성 분석)

  • Choi, Sun Do
    • Journal of Applied Biological Chemistry
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    • v.56 no.3
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    • pp.137-139
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    • 2013
  • Free radical scavengers in the bisdemethoxycurcumin (BDMC), demethoxycurcumin (DMC) and curcumin of turmeric (Curcuma longa) were screened, identified, quantified and isolation using coupled off-line-2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) and on-line screening high-performance liquid chromatography (HPLC)-ABTS assay. There was a very small margin of error between the off-line-ABTS method and the on-line screening HPLC-ABTS method.

Chemiluminescence immunochromatographic analysis for the quantitative determination of algal toxins

  • Pyo, Dongjin;Kim, Taehoon
    • ALGAE
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    • v.28 no.3
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    • pp.289-296
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    • 2013
  • For the quantitative detection of algal toxin, microcystin, a chemiluminescence immunochromatographic assay method was developed. The developed system consists of four parts, chemiluminescence assay strip (nitrocellulose membrane), horse radish peroxidase labeled microcystin monoclonal antibodies, chemiluminescence substrate (luminol and hydrogen peroxide), and luminometer. The performance of the chemiluminescence immunochromatographic assay system was compared with high performance liquid chromatography (HPLC) detection. The detection limit of chemiluminescence immunochromatographic assay system is several orders of magnitude lower than with HPLC. The chemiluminescence immunochromatography and HPLC results correlated very well with the correlation coefficient ($r^2$) of 0.979.

Analysis of Organic Acid in Korean Apple Juice by High Performance Liquid Chromatography (High Performance Liquid Chromatography에 의한 사과주스의 유기산 분석)

  • 황혜정;김성수;윤광로
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.2
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    • pp.181-187
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    • 2000
  • The contents of organic acid in Korean apple juice were analyzed by HPLC using YMC-peak ODS-AQ column and enzymatic assay. Model apple juices were prepared at the laboratory and commercial apple juices were purchased from the market. Individual organic acid contents were as follows: DL-malic acid 62~402mg%, L-malic acid 48~360mg%, citric acid 1.81~15.74mg%, fumaric acid nd~0.50mg%. Together, these tests gave useful information about the quality and authenticity of a particular apple juice smaple. The presence of D-malic acid was a clear indication of adulteration because this isomer did not occur naturally. Fumaric acid and citric acid levels above trace amounts were also inconsistent with pure apple juice.

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The Rapid Determination of Gemcitabine by Reversed-phase Ultra-Performance Liquid Chromatography (역상 초고속액체크로마토그라피에 의한 gemcitabine의 빠른 농도 분석법)

  • Park, Dae-Jin;Kim, Woo-Mi
    • Journal of Life Science
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    • v.19 no.12
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    • pp.1698-1704
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    • 2009
  • Gemcitabine is an anticancer drug used to treat a variety of solid tumors. The drug is rapidly inactivated by cytidine deaminase in plasma and its hydrophilicity restricts the extent of quantification that is possible using reversed-phase liquid chromatography. In this paper, we report a rapid and precise method to analyze velocity and peak efficiency using ultra-performance liquid chromatography (UPLC) with a reversed-phase column. The retention periods of gemcitabine and 2'-deoxycytidine at 283 nm were 3.2 and 2.1 min, respectively. The assay provided highly linear results in the range of $0.1{\sim}20{\mu}g/ml$ ($r^2$ > 0.999). The coefficients of variation of the intra-day and inter-day assays were less than 10.0%. We observed that the estimated average concentrations of the intra-day and inter-day assays ranged from 97.3 to 113.5% to verify the accuracy. These results suggest that this new reversed-phase UPLC method is a rapid and reliable way of determining gemcitabine levels.