• Title/Summary/Keyword: reversed-phase HPLC

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Seasonal Variation of Flavonoid Contents in the Leaves of Cedrela sinensis (계절에 따른 참죽나무잎 Flavonoid 화합물의 변화 분석)

  • 박종철;김성환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.4
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    • pp.578-581
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    • 1995
  • The seasonal variation of two flavonoid, quercitrin and afzelin, in the leaves of Cedrela sinensis A. Juss. from May until November was investigated by high performance liquid chromatography. Separation by reversed phase chromatography on ${\mu}-Bondapak\;C_{18}$ column was achieved by isocratic elution. The concentration of quercitrin was 5.81% (w/w) in May and high in July(8.89%, w/w), August(7.94%, w/w), and decreased from September to November. The content of another flavonoid, afzelin was 0.06% (w/w) in May and highest in June (0.12%, w/w), and was reduced from July.

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Rapid Separation and Indentification Method of Tea Catechins (녹차 중 카테킨류의 신속 분리 및 동정법)

  • Lee, Jeong-Hee;Lee, Yong-Moon;Moon, Dong-Cheul
    • Analytical Science and Technology
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    • v.5 no.3
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    • pp.333-338
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    • 1992
  • The tea tannins, epigallocatechin, epigallocatechin gallate, were successfully separated by a Sephadex LH-20 column by the acetone based gradient elution. Each fractions was collected by monitoring at 280nm. Purified fractions were directly characterized by fast atom bombardment mass spectrometry. Epigallocatechin and epigallocatechin gallate were identified and shown as low as 70% purity in the reversed phase column. This revised method is more advantageous than known methods in purity and rapidity.

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Analysis of the Urushiol in Korean Lacquer (한국산 옻칠의 우루시올 성분 분석)

  • Kim, Jung-Bae
    • The Korean Journal of Food And Nutrition
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    • v.19 no.3
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    • pp.267-270
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    • 2006
  • In Korea, for a long time Rhus verniciflua has traditionally been used as an herbal medicines plants. A stem of Rhus verniciflua has been used to treat gastrointestinal trouble with in form of boiled chicken as a folk medicine. But it has been recognized as an extremely active allergen causing skin reactions. The chief allergenic component, urushiol, is found within the oleoresinous sap of Rhus verniciflua. Most components of urushiol have unsaturated side chains. These unsaturated side chains of urushiol are important to polymerization of these natural products. The urushiol components in Korean lacquer were isolated by reversed phase HPLC. The molecular weight of purified urushiol was determined as 340 from mass analysis. This compound was identified as Heptadecatetraenyl catechol (MW 340).

HPLC SEPARATION AND QUANTITATIVE DETERMINATION OF GINSENOSIDES FROM PANAX GINSENG, PANAX QUINQUEFOLIUM AND FROM GINSENG DRUG PREPARATIONS

  • Soldati F
    • Proceedings of the Ginseng society Conference
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    • 1980.09a
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    • pp.59-69
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    • 1980
  • A new HPLC-method for separation and quantitative determination of ginsenosides in Panax ginseng, Panax quinquefolium and in pharmaceutical drug preparations is elaborated. A reversed-phase-system with ${\mu}Bondapak\;C_{18}$ column (3.9 mm $I.D.{\times}30\;cm$) using acetonitrile-water (30:70) 2 ml/min and acetonitrile-water (18:82) 4 ml/min is suitable for the base-line separation of $Rb_1,\;Rb_2,\;Rc,\;Rd,\;Rf,\;Rg_2,\;respectively\;Re,\;Rg_1$ in 30 minutes. The ginsenosides are directly detected at 203 nm (without derivatization) with the LC-55 or LC-75 spectrophotometer (Perkin-Elmer) at $100\%$ transmission. Detection limit is 300 ng at a signal-to-noise ratio of 10:1. The ginsenosides-peak identification is carried out with HPTLC (high performance thin layer chromatography), with MIR-IR (multiple internal reflection-IR-spectros-copy) and with FD-MS (field desorption mass spectrometry). The calibration curve of each ginsenoside has a correlation coefficient very near to 1. Relative standard deviation for quantitative determinations depends upon the amount of ginsenosides and is approximately 1\%$ for ginsenoside contents of 1\%$. This method is adaptable for routine analysis in quality control laboratories.

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Two Biophenolic Glycosides from Portulaca oleracea (쇠비름에서 분리된 2개의 Biophenolic Glycosides)

  • Youngwan Seo;Jongheon Shin;Burm Jong Lee;Dong Seok Lee
    • Journal of the Korean Chemical Society
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    • v.47 no.1
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    • pp.43-46
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    • 2003
  • From Portulaca oleracea which is widely distributed in Korea and has long been used as a folk medicine, two biophenolic glycosides, 3-hydroxy-1-(2-hydroxyethyl)phenyl-4-O-${\beta}$-D-glucopyranoside (1) and 2-(3,4-dihydroxyphenyl) ethyl-O-${\beta}$-D-glucopyranoside (2) were isolated using column chromatography and reversed-phase HPLC. $^{13}C$ NMR spectral assignment for these compounds was revised by the extensive 2-D NMR experiments such as NOESY, HMQC, and HMBC. These compounds showed a considerable antioxidant effect in DPPH assay system.

Determination of Recombinant Human Epidermal Growth factor (rhEGF) in a Pharmaceutical Formulation by High Performance Liquid Chromatography with Electrochemical Detection

  • Lee, Kang-Woo;Hwang, Kyung-Hwa;Kim, Chang-Soo;Han, Kun;Chung, Youn-Bok;Park, Jeong-Sook;Lee, Yong-Moon;Moon, Dong-Cheul
    • Archives of Pharmacal Research
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    • v.24 no.4
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    • pp.355-359
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    • 2001
  • A novel HPLC method with electrochemical detection has been developed for the determination of recombinant human epidermal growth factor (rhEGF) in pharmaceutical products. rhEGF was separated from other components in formulation on a reversed-phase C18 column with 24% acetonitrile in 0.1 M phosphate buffer (pH 4.75). The optimum electrochemical oxidation of EGF was obtained at 0.85 V vs. Ag/AgCl in a glassy carbon working electrode due to electroactive tyrosine, tryptophan, methionine, and arginine residues. The quantitation range was from 1.0 to 200 ng of rhEGF with the linear correlation coefficient greater than 0.999. The method was successfully applied for the quantitation of rhEGF in a pharmaceutical preparation.

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Liquid Chromatographic Determination of Etofenprox Residues in Foods with Mass-Spectrometric Confirmation

  • Lee, Young-Deuk;Kwon, Chan-Hyeok;Kwon, Ki-Sung
    • Korean Journal of Environmental Agriculture
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    • v.30 no.4
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    • pp.432-439
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    • 2011
  • BACKGROUND: An official analytical method was developed to determine etofenprox residues in agricultural commodities using high-performance liquid chromatography (HPLC). METHODS AND RESULTS: The etofenprox residue was extracted with acetone from representative samples of five raw products which comprised rice grain, apple, mandarin, cabbage, and soybean. The extract was then serially purified by liquid-liquid partition and Florisil column chromatography. For rice and soybean samples, acetonitrile/n-hexane partition was additionally coupled to remove nonpolar lipids. Reversed phase HPLC using an octadecylsilyl column was successfully applied to separate etofenprox from co-extractives. Intact etofenprox was sensitively detected by ultraviolet absorption at 225 nm. Recovery experiment at the quantitation limit validated that the proposed method could apparently determine the etofenprox residue at 0.02 mg/kg. Mean recoveries from five crop samples fortified at three levels in triplicate were in the range of 93.6~106.4%. Relative standard deviations of the analytical method were all less than 10%, irrespective of crop types. A selected-ion monitoring LC/mass spectrometry with positive atmospheric-pressure chemical ionization was also provided to confirm the suspected residue. CONCLUSION(s): The proposed method is simple, rapid and sensitive enough to be employed in routine inspection or monitoring of agricultural products for the etofenprox residue.

Antioxidant Activities and Determination of Phenolic Acids from Leaves of Perilla frutescens

  • Lee, Jin-Hwan;Kang, Nam-Suk;Ha, Tae-Joung;Ko, Jong-Min;Han, Won-Young;Suh, Duck-Yong;Park, Ki-Hun;Baek, In-Youl
    • Journal of Applied Biological Chemistry
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    • v.49 no.1
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    • pp.11-15
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    • 2006
  • Two catecholic phenolic acids (1 and 2) obtained from 80% methanolic extract of Perilla frutescens var. frutescens leaves through chromatography showed strong DPPH (1: $IC_{50}$ = 15.5 ${\mu}M$, 2: $IC_{50}$ = 11.7 ${\mu}M$) and ABTS (1: $IC_{50}$ = 5.5 ${\mu}M$, 2: $IC_{50}$ = 4.6 ${\mu}M$) radicals scavenging abilities. Antioxidant compounds contents of 1 and 2 as determined by $C_{18}$ reversed phase HPLC coupled with diode-array detector were 2.98 and 2.26 mg/g, respectively.

Determination of ${\alpha}-Keto$ Acids in Serum and Urine Using 1,2-Diamino-4,5-methylendioxybenzene as a Fluorescent Derivatizating Agent by High Performance Liquid Chromatography (HPLC법에 의한 1,2-디아미노-4,5-메틸렌디옥시벤젠을 형광유도체화제로 한 혈청 및 뇨 중의 ${\alpha}$-케토산의 분석)

  • Ok, Chi-Wan;Kim, Dae-Ki;Park, Song-Ja;Park, Jong-Sei
    • YAKHAK HOEJI
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    • v.36 no.4
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    • pp.370-378
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    • 1992
  • A simple and sensitive high performance liquid chromatographic method to quantitate ${\alpha}-keto$ acids in serum and urine was investigated. ${\alpha}-Keto$ acids react with 1,2-diamino-4,5-methylenedioxybenzene (DMB) in the presence of 2-mercapto-ethanol and sodium hydrogen sulfite to form highly fluorescent derivatives, substituted 6,7-methylenedioxyquinoxalinol. The derivatization procedure was performed in water bath at $100^{\circ}C$, and completed within 50 min. By the use of a reversed-phase column and multi-step gradient with two solvents, a mixture containing twelve of these derivatives were efficiently resolved within 35 minutes. The optimal wavelengh of the fluorescence detector are ${\lambda}_{ex}=364\;nm$ and ${\lambda}_{em}=445\;nm$. The quantitation of the individual ${\alpha}-Keto$ acids was reproducible with relative standard deviation of $3.0{\sim}7.9%$ and had a detection limits of $10{\sim}60$ fmol, except for p-hydroxyphenylpyruvic acid (960 fmol).

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Quantitative Analysis of Antioxidants in Sesame Seed (참깨 종실의 항산화 성분 정량분석 연구)

  • Ryu, Su-Rho;Lee, Jung-Il;Kang, Sam-Sik;Choi, Chang-Yoel
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.4
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    • pp.377-382
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    • 1992
  • This study was conducted to obtain basic informations on the lignan components from sesame seed. Two major lignans, sesamin and sesamolin, were isolated and identified by means of spectral methods, and quantitative analysis was by HPLC from sesame variety Danbaeggae. Separation was achieved by isocratic elution and reversed phase chromatography Develosil ODS column. The content of the major lignan components were about 0.42% and 0.30% for sesamin and sesamolin, respectively.

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