• Title/Summary/Keyword: Reversed phase liquid chromatography

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Determination of Aloesin in Plasma by High-Performance Liquid Chromatography as Fluorescent 9-Anthroyl Derivative

  • Kim, Kyeong-Ho;Lee, Jin-Gyun;Park, Jeong-Hill;Shin, Young-Geun;Lee, Seung-Ki;Cho, Tae-Hyung
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.651-656
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    • 1998
  • A sensitive high-performance liquid chromatographic (HPLC) method for the determination of aloesin in plasma was developed. After solid-phase extraction from plasma and derivatization of aloesin and compound AD-1, which was prepared from aloesin as a internal standard, with 9-anthroylnitrile in the presence of quinuclidine, the derivatives were separated on a Inertsil ODS-3 column using acetonitrile/methanol/water (3:1:6) as a mobile phase, and detected fluorimetrically at 460nm with excitation at 360nm. The detection limit of aloesin was 3.2ng/ml in plasma (S/N=3).

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Quantitative Determination of Salidroside and Tyrosol from the Underground Part of Rhodiola rosea by High Performance Liquid Chromatography

  • Linh, Pham-Tuan;Kim, Young-Ho;Hong, Seon-Pyo;Jian, Jiang-Ji;Kang, Jong-Seong
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.349-352
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    • 2000
  • A reversed-phase high performance liquid chromatographic method was developed to determine salidroside and tyrosol simultaneously in the Rhodiola rosea. The optimum condition was Nova-pak $C_18$as stationary phase, 6.5% methanol in water as mobile phase and detection at UV 225 nm. The identification was carried out by comparing the retention time and LC/MS spectrum of the relevant peaks with those of isolated standards. The contents of salidroside and tyrosol in the samples gathered from various area in China were ranged over 1.3-11.1 ${m}g/g$ and 0.3-2.2 ${m}g/g$, respectively.

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Quantitative Analysis of Lovastatin in Human Plasma and urine by Reversed-Phase High-Performance Liquid Chromatography (역상 고속액체크로마토그라프법을 이용한 혈장 및 뇨 중 로바스타틴의 정량)

  • Choi, Hye-Jin;Kim, Myoung-Min;Choi, Kyung-Eob
    • YAKHAK HOEJI
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    • v.42 no.5
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    • pp.473-479
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    • 1998
  • Lovastatin (LOVA), a fungal metabolite isolated from cultures of Aspergillus terreus, is a competitive HMG-CoA reductase inhibitor used for the treatment of primary hyper cholesterolemia, and has also been shown to suppress growth in a variety of non-glioma tumor cell lines. A sensitive reversed-phase high-perfonnance liquid chromatographic method with ultraviolet (UV) absorbance detection has been developed to quantitate LOVA in human plasma and urine samples using liquid-liquid extraction procedure. Baseline separation of LOVA and internal standard, simvastatin was achieved on a Novapak $C_{18}$ analytical column with a mobile phase containing 0.025M $NaH_2PO_4$: CAN (35:65, v/v%), adjusted pH to 4.5. The flow rate was set at 1.5ml/min, and the column effluent was monitored by a UV detection at 238nm. The limit of quantification was determined to be 0.5${\mu}$g/ml while extraction efficiency of LOVA ranged from 73.4-82.9% at LOVA concentrations of 0.5 to 10${\mu}$g/ml. Good linearity with correlation coefficients greater than 0.999 was obtained in the range of LOVA concentrations from 0.5 to 10${\mu}$g/ml. The accuracy and the precision were proven excellent with relative standard deviation (RSD, %) and relative error (RE, %) of less than 4.2 and 4.0, respectively. Intraday precision, evaluated at five LOVA concentrations (0.5, 1, 2, 5, 10${\mu}$g/ml) and expressed as RSD ranged from 0-1.82% while the interday precision at the same concentrations ranged from 0.7-10.5%. The analytical method described was then successfully employed for the determination of LOVA concentrations in plasma samples obtained during a phase II clinical trial using high doses of LOVA (30-40mg/kg/day). This method could be further utilized for the ongoing pharmacolkinetic studies and therapeutic drug monitoring of the high-dose LOVA therapy in adenocarcinoma patients.

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Quantitative Determination of Psoralen and Angelicin from Some Medicinal Herbs by High Performance Liquid Chromatography

  • Dong, Nguyen-Thanh;Bae, Ki-Hwan;Kim, Young-Ho;Hwang, Gwi-Seo;Heo, Ok-Soon;Kim, Se-Eun;Kang, Jong-Seong
    • Archives of Pharmacal Research
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    • v.26 no.7
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    • pp.516-520
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    • 2003
  • A reversed-phase high performance liquid chromatographic method was developed to determine the contents of psoralen and angelicin from some medicinal herbs. The optimum eluent for chromatography was 20 v/v% acetonitrile in water on a Zorbax 300SB $C_{18}$ column. The identification was carried out by comparing the retention time and mass spectra of the relevant peaks with their standards. The variation of the concentration of psoralen and angelicin was wide between different species. The seeds of Psoralea corylifolia showed the highest contents of psoralen (7.8 mg/g) and angelicin (2.3 mg/g) among the tested herbs.

Quantitative analysis of cordycepin in Cordyceps militaris under different extraction methods

  • Choi, Jungwon;Paje, Leo Adrianne;Kwon, Baekjun;Noh, Jaekyu;Lee, Sanghyun
    • Journal of Applied Biological Chemistry
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    • v.64 no.2
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    • pp.153-158
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    • 2021
  • Cordyceps militaris (CM) is one of the most important medicinal mushrooms known to possess various biological activities. Cordycepin (CP) is a bioactive compound present in the fruiting bodies of CM and is known to have anti-tumor, anti-metastatic immunomodulatory and anti-inflammatory activities. In this study, we aim to analyze CP quantitatively under various CM extraction conditions. CP was measured using high-performance liquid chromatography, quantified using a reversed phase column using a gradient elution system of water and acetonitrile, and detected with a UV absorbance wavelength of 260 nm. The CP content of CM was the highest in 100% ethanol extract of the fruiting bodies and 60% ethanol extract of the mycelium. This study provides an efficient analysis method to determine the optimal extraction conditions for CP that can be used as a basis for developing functional foods and pharmaceutical products derived from CM.

Separation and Detection of Nonchromophore Aliphatic Compounds by Reversed-Phase Liquid Chromatography using Ultraviolet-Absorbing Reagent (자외선 흡수물질을 이용한 역상 액체 크로마토그라피에 의한 비흡수 지방족 화합물들의 검출과 분리)

  • Lee Seung-Seok;Kang Sam-Woo;Oh Hae-Beom
    • Journal of the Korean Chemical Society
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    • v.35 no.4
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    • pp.397-404
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    • 1991
  • Nonchromophore compounds such as aliphatic acids, alcohols and tetraalkylammonium salts could be detected by indirect photometric detection on the revered-phase liquid chromatography. Benzyltriethylammonium bromide(BTEAB) was used as a detection reagent. Also, the retention mechanism and response of samples were investigated to the several factors such as pH, temperature, and concentration of MeOH as well as concentration of detection reagents in mobile phase. And some mixture of samples were able to be separated under optimum condition.

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Comparison and Estimation of Equilibrium Constants for Deoxyribonucleosides by Plate Theory and Moment Method (단이론과 모멘트방법을 이용한 데옥시리보뉴클레오사이드의 평형상수의 계산 및 비교)

  • Lee, Ju Weon;Row, Kyung Ho
    • Applied Chemistry for Engineering
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    • v.8 no.3
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    • pp.403-409
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    • 1997
  • The equilibrium constants of five deoxyribonucleosides (dDyd, dUrd, dGuo, dThd, dAdo) were estimated by the plate theory and the moment method under isocratic conditions of the Reversed-Phase High Performance Liquid Chromatography (RP-HPLC). The mobile phase in this system was composed of water and organic modifiers(acetonitrile and methanol) The plate theory of linear adsorption isotherm was treated on the basis of continuous flow of eluent through the plates of the column. The moment method was utilized to find the equilibrium constant from the first absolute moment of experimental data. The equilibrium constants of five deoxyribonucleosides in the two methods were very close, and also the equilibrium constants calculated by capacity factor were similar to those by both the plate theory and the moment method. The equilibrium constant was expressed as a semi-log function of the quantity of organic modifier. Excellent agreements between the calculated elusion profile by the plate theory and the experimental data were observed.

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Determination of Glimepiride in Human Plasma by Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry

  • Kim, Ho-Hyun;Chang, Kyu-Young;Lee, Hee-Joo;Han, Sang-Beom
    • Bulletin of the Korean Chemical Society
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    • v.25 no.1
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    • pp.109-114
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    • 2004
  • A sensitive method for quantitation of glimepiride in human plasma has been established using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Glipizide was used as an internal standard. Glimepiride and internal standard in plasma sample was extracted using diethyl etherethyl acetate (1 : 1). A centrifuged upper layer was then evaporated and reconstituted with the mobile phase of acetonitrile-5 mM ammonium acetate (60:40, pH 3.0). The reconstituted samples were injected into a $C_{18}$ reversed-phase column. Using MS/MS in the multiple reaction monitoring (MRM) mode, glimepiride and glipizide were detected without severe interference from human plasma matrix. Glimepiride produced a protonated precursor ion ([M+H]$^+$) at m/z 491 and a corresponding product ion at m/z 352. And the internal standard produced a protonated precursor ion ([M+H]]$^+$) at m/z 446 and a corresponding product ion at m/z 321. Detection of glimepiride in human plasma by the LC-ESI/MS/MS method was accurate and precise with a quantitation limit of 0.1 ng/mL. The validation, reproducibility, stability, and recovery of the method were evaluated. The method has been successfully applied to pharmacokinetic studies of glimepiride in human plasma.

Recent Developments in High-performance Liquid Chromatography of Lipids

  • Christie, William W.
    • Journal of the Korean Applied Science and Technology
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    • v.10 no.1
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    • pp.1-8
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    • 1993
  • The possibilities for HPLC analysis of lipids have been revolutionised by the availability of evaporative light-scattering detectors, with which the response is independent of the nature of the mobile phase and does not depend On the presence of specific chromophores in the lipids. It was thus possible to develop an HPLC procedure, involving ternary gradient elution, for separating all the lipid classes in animal tissues in a single step. Although reversed-phase HPLC has been widely used for the analysis of molecular species of lipids, sliver ion chromatography can be a valuable alternative. For example, a stable silver ion column for HPLC was developed which permitted resolution of molecular species of triacylglycerols, even from such complex samples as fish oils, again With light-scattering detection and gradient elution. The capacity for HPLC resolution of diastereomeric diacyl-sn-glycerol derivatives, prepared from triacylglycerols. has lead to a new simple method for stereospecific analysis of the latter.