色谱

色谱

• 研究论文 • 上一篇    下一篇

高效液相色谱法测定大鼠血浆中的原儿茶酸

韩瑛,熊志立,杨春娟,刘曼,李发美   

  1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China
  • 收稿日期:2006-07-17 修回日期:2006-10-25 出版日期:2007-03-30 发布日期:1985-12-25
  • 通讯作者: 李发美

Determination of Protocatechuic Acid in Rat Plasma by High Performance Liquid Chromatography

HAN Ying, XIONG Zhili, YANG Chunjuan, LIU Man, LI Famei   

  1. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China
  • Received:2006-07-17 Revised:2006-10-25 Online:2007-03-30 Published:1985-12-25
  • Contact: LI Famei

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摘要： 建立了大鼠血浆中原儿茶酸含量测定的高效液相色谱方法。采用的色谱柱为DiamondsilTM C18 柱(150 mm×4.6 mm,5 μm);流动相为乙腈-水(体积比为9∶91,用H3PO4 调pH至2.5);流速1.2 mL/min;检测波长260 nm;内标为对羟基苯甲酸。原儿茶酸的线性范围为0.050~3.20 mg/L,线性相关系数为0.9978，最低定量限为0.050 mg/L,日内和日间测定的精密度（以相对标准偏差表示）均低于7.0%,准确度（以相对误差表示）为-1.4%～2.6%；在0.050,0.40,3.20 mg/L低、中、高3个添加浓度水平下，血浆样品的提取回收率分别为83.4%,87.3%,91.1%。该方法简便,灵敏,准确,适用于大鼠体内原儿茶酸的药物动力学研究。

关键词: 大鼠血浆 , 高效液相色谱法, 原儿茶酸

Abstract: A method was developed for the quantitative determination of protocatechuic acid in rat plasma by high performance liquid chromatography (HPLC). With added p-hydroxybenzoic acid as internal standard, the plasma samples were extracted with a solvent mixture of methanol and acetonitrile (1∶5, v/v). The analyte was determined by HPLC with a DiamondsilTM C18 column and a mobile phase of acetonitrile-water (adjusted to pH 2.5 with H3PO4 , 9∶91, v/v) at a flow rate of 1.2 mL/min. The UV detection wavelength was 260 nm. The assay exhibited a good linearity in the concentration range from 0.050 to 3.20 mg/L with a correlation coefficient of 0.9978. The limit of quantification was 0.050 mg/L. The relative standard deviations of  intra-day and inter-day determination  were both less than 7.0% and the accuracy ranged from -1.4% to 2.6%. The extraction recoveries of protocatechuic acid  from rat plasma samples at three concentration levels were 83.4%, 87.3%, and 91.1%, respectively. The developed method was applied to a pharmacokinetic study. The main pharmacokinetic parameters in rats after a single oral dose of 10 mL/kg body weight were as follows: Cmax of (3.16±0.03) μg/mL, tmax of (0.50±0.00) h, AUC0t of (39.9±5.9) μg·mL-1·h, AUC0∞ of (54.8±8.1) μg·mL-1·h and t1/2 of (3.87±0.25) h. The method was proved to be simple, sensitive and accurate, thus suitable for the pharmacokinetic study of protocatechuic acid in rat plasma.

Key words: protocatechuic acid, rat plasma , high performance liquid chromatography (HPLC)