Simultaneous determination of phenylglyoxylic acid, mandelic acid, styrene glycol and hippuric acid in primary culture of rat hepatocytes incubate by high-performance liquid chromatography
Simultaneous determination of phenylglyoxylic acid, mandelic acid, styrene glycol and hippuric acid in primary culture of rat hepatocytes incubate by high-performance liquid chromatography
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DOI：
10.1002/bmc.783
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发表时间：
2007-05-01
影响因子：
1.8
通讯作者：
Zeng, Su
中科院分区：
文献类型：
--
作者：
Wang, Jin-Zhao;Lu, Xin-Yan;Zeng, Su
建立了同时测定细胞培养液中苯乙醛酸（PGA）、扁桃酸（MA）、苯乙二醇（SG）和马尿酸（HA）的高效液相色谱法。在C1色谱柱上进行分析，色谱柱移动的流动相由甲醇-磷酸二氢钾（pH 2.5; 10 mM; 10：90，v/v）组成，检测波长为220 nm。移动的流速为0.5 mL/min，PGA、MA、SG和HA的平均绝对回收率分别为95.9%、98.4%、98.0%和97.1%。在三个浓度水平下测定的日间和日内精密度均小于RSD的10%。PGA、MA、SG和HA的定量限分别为13.2、13.1、14.5和11.2 μ M，RSD均小于20%。PGA、MA、SG和HA的检测限分别为4.6、4.6、5.1和3.9 μ M。该方法已成功地应用于研究SG和MA在原代培养大鼠肝细胞中的立体选择性代谢。结果表明，MA和SG在原代培养的大鼠肝细胞中均存在立体选择性代谢。从S-MA到PGA的生物转化程度显著大于从R对映体的生物转化程度，并且S-SG和R-SG的主要代谢产物分别是PGA和HA。版权所有（c）2007约翰威利父子有限公司。
A simple HPLC method for the simultaneous determination of phenylglyoxylic acid (PGA), mandelic acid (MA), styrene glycol (SG) and hippuric acid (HA) in cell culture medium was developed. Analysis was performed on a C, column with a mobile phase composed of methanol-potassium dihydrogen phosphate (pH 2.5; 10 mM; 10:90, v/v) at 220 nm. The flow-rate of mobile phase was set at 0.5 mL/min. The mean absolute recoveries of PGA, MA, SG and HA were 95.9, 98.4, 98.0 and 97.1 %, respectively. The inter-day and intra-day precisions, determined at three concentration levels, were less than 10% of RSD. The limits of quantification for PGA, MA, SG and HA were 13.2, 13.1, 14.5 and 11.2 mu M with RSD less than 20%. The limits of detection for PGA, MA, SG and HA were 4.6, 4.6, 5.1 and 3.9 mu M, respectively. The method was successfully applied to study the stereoselective metabolism of SG and MA in primary culture of rat hepatocytes. The results show that there is stereoselective metabolism for both of MA and SG in primary culture of rat hepatocytes. The extent of biotransformation from S-MA to PGA is significantly greater than that from the R enantiomer and the main metabolites are PGA and HA for S-SG and R-SG, respectively. Copyright (c) 2007 John Wiley & Sons, Ltd.