• Title/Summary/Keyword: HPLC Chromatography

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Simultaneous Determination of Cinnamaldehyde and Coumarin in Oryeong-san using HPLC with Photodiode Array Detector

  • Seo, Chang-Seob;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.18 no.2
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    • pp.251-257
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    • 2010
  • Objectives : To develop and validate High-performance liquid chromatography-photodiode array methods for simultaneous determination of two constituents in Oryeong-san(ORS). Methods : Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array(PDA) detection at 280 nm, were used for quantification of the two marker components of ORS. The mobile phase using a gradient flow consisted of two solvent systems. Solvent A was $H_2O$ and solvent B was acetonitrile. Results : Calibration curves were acquired with correlation coefficient ($r^2$)>0.9999, and the relative standard deviation(RSD) values(%) for intra- and inter-day precision were not exceed 1.0%. The recovery rate of each compound was in the range of 93.01-104.16%, with an RSD less than 2.0%. The contents of two compounds in ORS were 1.10-3.72 mg/g. Conclusions : The established HPLC method will be helpful to improve quality control of ORS.

Multiresidue Analysis of Eight Acaricides in Fruits

  • Lee, Young-Deuk;Kwon, Chan-Hyeok
    • Journal of Applied Biological Chemistry
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    • v.42 no.4
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    • pp.191-196
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    • 1999
  • A multiresidue analytical method was developed for eight acaricides including benzoximate, clofentezine, fenazaquin, fenothiocarb, fenpyroximate, hexythiazox, pyridaben, and tebufenpyrad in four major fruits using high-performance liquid chromatography (HPLC). All the confounds were extracted with acetone from apple, pear, grape, and citrus samples. The extract was diluted with saline water, and n-heaxane partition was followed to recover the acaricides. Florisil column chromatography was employed to further purify the sample extract. HPLC with ultraviolet absorption detection, using an octadecylsilyl column under the isocratic mobile phase of acetonitrile/water mixture, was successfully applied to separate and quantitate all the compounds in the purified extract. Recoveries of the eight acaricides from for fortified samples ranged 86.4~97.0%. Relative standard deviations of the analytical method were all less than 10%. Detection limits of the method were in the range of 0.02~0.05 mg/kg. The proposed method was reproducible and sensitive enough to evaluate the terminal residue of the eight acaricides in the fruit harvest.

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Rapid Method for Seperation and Quantitation of p-Nitrophenol Derivative by HPLC (p-Nitrophenol 유도체의 HPLC에 의한 신속 분리 정량)

  • 이완구
    • Journal of Environmental Health Sciences
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    • v.9 no.1
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    • pp.89-93
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    • 1983
  • Various analytical methods for p-Nitrophenol derivatives have been reported as follows. 1) Thin-Layer Chromatography, 2) Gas Chromatography, 3) Cholinesterase Activity Determination, 4) Diazo Method, 5) Nitrophenol Method, 6) Indophenol Method. But these methods are mainly analyse total quantity of p-Nitrophenol and are not available for the seperation and pose some analytical problems associated with extensive clean up procedure. A rapid and simple method was developed for the seperation and quantitation for the p-Nitrophenol and it's derivatives by HPLC. Also an experiment was undertaken by the authors for the quantitation of the p-Nitrophenol in the blood of the intoxicated body. Levels of p-Nitrophenol ranging from approximately 0.10 to $1.69 \mu g/ml$ for Parathion and $3.44 \mu g/ml$ for EPN in each sample were measured with the average recovery of $95.5\pm0.52%$

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Temperature Controllable HPLC Column for Preparative Fractionation of Polymers

  • Im, Kyu-Hyun;Park, Hae-Woong;Kim, Young-Tak;Chang, Tai-Hyun
    • Macromolecular Research
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    • v.16 no.6
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    • pp.544-548
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    • 2008
  • An HPLC column with a self-contained temperature control device was constructed for preparative temperature programmed interaction chromatography. Two Peltier plates were attached to a large bore column ($120{\times}22\;mm$ i.d.) and the column temperature was controlled by PID mode feed back control. At a flow rate of 1.5 mL/min, the column temperature could be increased and decreased at a rate as high as $50^{\circ}C/min$ and $10^{\circ}C/min$, respectively, which is much faster than using a column jacket and bath/circulator. The rapid heating and cooling rates allows a high repetition rate of chromatographic fractionation. The performance of the temperature controllable column was demonstrated successfully by the fractionation of homo-polymer precursors from diblock copolymers.

Determination of isovitexin from Lespedeza cuneata using a validated HPLC-UV method

  • Lee, Ju Sung;Paje, Leo Adrianne;Yoo, Sang-Woo;Lee, Seong;Ku, Ja-Jung;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • v.64 no.1
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    • pp.63-67
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    • 2021
  • Isovitexin, a marker compound with various pharmacological activities, in Lespedeza cuneata, was analyzed using high performance liquid chromatography coupled with UV (HPLC/UV). There are no previous reports on using L. cuneata as the source material for the quantification of isovitexin. In this study, we developed an optimized method using HPLC-UV analysis, which was validated using various parameters. Our method demonstrated high specificity, and good separation of the chromatographic peak was achieved. Parameters such as linearity (r2≥0.9997), precision, and accuracy indicated that our proposed analytical method had good reliability and sensitivity. These results demonstrate the utility and convenience of our method for rapidly quantifying isovitexin in L. cuneata extracts.

Transformation of dissolved organic matter in a constructed wetland: A molecular-level composition analysis using pyrolysis-gas chromatography mass spectrometry

  • Park, Jongkwan;Choi, Mijin;Cho, Jaeweon;Chon, Kyongmi
    • Environmental Engineering Research
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    • v.23 no.4
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    • pp.390-396
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    • 2018
  • This study investigated the transformation of dissolved organic matter (DOM) in a free-water surface flow constructed wetland. Pyrolysis gas chromatography-mass spectrometry (Py-GC/MS) coupled with preparative high-performance liquid chromatography (prep-HPLC) was used to analyze the compositions of biopolymers (polysaccharides, amino sugars, proteins, polyhydroxy aromatics, lipids and lignin) in DOM according to the molecular size at three sampling points of the water flow: inflow, midflow, and outflow. The prep-HPLC results verified the decomposition of DOM through the decrease in the number of peaks from three to one in the chromatograms of the sampling points. The Py-GC/MS results for the degradable peaks indicated that biopolymers relating to polysaccharides and proteins gradually biodegraded with the water flow. On the other hand, the recalcitrant organic fraction (the remaining peak) in the outflow showed a relatively high concentration of aromatic compounds. Therefore, the ecological processes in the constructed wetland caused DOM to become more aromatic and homogeneous. This indicated that the constructed wetland can be an effective buffer area for releasing biochemically stable DOM, which has less influence on biological water quality indicators, e.g., biochemical oxygen demand, into an aquatic ecosystem.

Comparison of Methods for Measuring Histamine by ELISA and HPLC-MS Assay In Vitro (In Vitro에서 히스타민 측정 시 ELISA법과 HPLC-MS 분석법의 비교)

  • Lee, In Hee;Kim, Yoo Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.306-312
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    • 2015
  • The measurement of histamine is to determine the degree of allergy because the allergic reaction can lead to the release of histamine. In general, the antigen-antibody reaction was quantified by measuring absorbance using a microplate reader. In this study, we compare the method using a general antigen-antibody reaction and the method using a high performance liquid chromatography mass spectrometer (HPLC-MS) of chemical analysis in the measurement of histamine secretion. The cell line used was RBL-2H3, an allergic reaction was induced by stimulation with C48/80 (compound 48/80). Allergy-induced cells degranulation rate was confirmed by measurement of ${\beta}$-hexosaminidase and cytotoxicity was performed for the validity of the experiment. The quantitative determination of histamine showed a significant difference, since the quantitative limit of the measurement by the antigen-antibody reaction was 10.257 ppm while the quantitative limit of the measurement by HPLC-MS was 0.020 ppm. Measurement of histamine in allergic activity and anti-allergy tests showed that the HPLC-MS analysis rather than the analysis of the antigen-antibody reaction is a more precise and accurate test.

Evaluation of Grinding Effects on the Extraction of Photosynthetic Pigments for HPLC Analysis (광합성 색소의 HPLC 분석을 위한 여과지 분쇄 효과 평가)

  • Jang, Su Jin;Park, Mi Ok
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.20 no.2
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    • pp.71-77
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    • 2015
  • High-Performance Liquid Chromatography (HPLC) is a widely used method for measuring the concentration of chlorophyll a as an indicator for estimating phytoplankton biomass and primary production and also for identifying carotenoids to determine phytoplankton composition. However, tissue grinding procedure requires a lot of time and experience in the analysis of multiple sample. Accordingly, we measured the concentrations of photosynthetic pigments before and after the grinding, in order to understand the grinding effects on the quantitative analysis of chlorophylls and carotenoids using samples from southwestern East Sea. When tissue grinding procedure was omitted, we found that Chl a concentrations were underestimated up to 45% in average. Also, concentrations of Zeaxanthin, 19'-butanoyloxyfucoxanthin, 19'-hexanoyloxyfucoxanthin, biomarkers of pico and nano-size phytoplankton, were underestimated up to maximum 77~85% without grinding. We found that the smaller the phytoplankton, the bigger underestimation of their biomarker pigments concentration is likely to happen due to the incomplete extraction. Thus, tissue grinding procedure should be included for HPLC analysis in all cases, to prevent the underestimation of not only Chl a but also carotenoids pigments.

Biosynthesis of $17\alpha-hydroxy$, $20\alpha-dihydroprogesterone$ by Ovaries of the Spotted Flounder (Verasper variegatus)

  • Baek Hea-Ja
    • Fisheries and Aquatic Sciences
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    • v.4 no.2
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    • pp.70-74
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    • 2001
  • To examine the production of steroids with potential oocyte maturation-inducing activity in the spotted flounder, Verasper variegatus, we have incubated post-vitellogenic oocytes (0.82­0.95mm in diameters) with radiolabeled pregnenolone and $17\alpha-hydroxyprogesterone$. The resulting metabolites were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The two main metabolites (progestogens) found in both incubations co-migrated with $17\alpha-hydroxy$, $20\alpha-dihydroprogesterone$ $(17\alpha, 20\alpha OHP)$ and $17\alpha-hydroxy,\;$20\beta-dihydroprogesterone$ (17 a20{30HP). Additional chromatography by HPLC and TLC confirmed the presence of radioactive $17\alpha, 20\alpha OHP$ and a large amount of unknown metabolite. The present study did not reveal in vitro formation of $l7\alpha 20\beta OHP$. Although 1$l7\alpha 20\beta OHP$ was found in a small amount, the synthesis of this steroid suggests that it may play a role in regulating the oocyte maturation process in the spotted flounder.

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Identification of Antimutagenic Compound from Kale by High Performance liquid Chromatography and Mass Spectrometry

  • Lee, Seon-Mi;Rhee, Sook -Hee;Yoo, Jong-Shin;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.3 no.4
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    • pp.334-338
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    • 1998
  • Kale(Brassica oleracea var. acephala) is one of Cruciferous vegetables that is closely related to the wild ancestral form of cabbabe. The ethanol extract of kale which contains the active compoundsss under Salmonella assay system was fractionated with chloroform to collect the nonpolar solvent soluble compounds, and then further fractionation was carried out by silica gel column chromatography. Among kale extracts separated by silical gel column chromatography, the fractions of 4, 5 and 6 exhibited strong antimutagenic activities. The major active compounds from the fraction were identified as chlorophyll derivatives by the analysis with HPLC-fritp-MS. The molecular weights of each chlorophyll derivatives in the sample were acquired from the peaks of positive ion atomosphere pressure chemical ionization (APCI) mas spectrometry.

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