使用基于蛋白质的手性固定相对手性酸在生物样品中的对映体选择性分析。
Enantioselective analysis of ritalinic acids in biological samples by using a protein-based chiral stationary phase.
作者信息
Zhang Jianhua, Deng Yulin, Fang Jim, McKay Gordon
机构信息
School of Life Science and Technology, Beijing Institute of Technology, No. 5, Zhongguancun St., South Beijing, 100081, China.
出版信息
Pharm Res. 2003 Nov;20(11):1881-4. doi: 10.1023/b:pham.0000003389.77585.be.
PURPOSE
This study was to develop and validate a new chiral HPLC-UV method for the quantitative analysis of enantiomeric ritalinic acid (RA) in human plasma.
METHODS
An alpha1-acid glycoprotein column was used with the mobile phase containing 0.4% acetic acid and 0.1% dimethyloctylamine, pH 3.4. The detection of enantiomeric RAs was at 220 nm.
RESULTS
A baseline separation for d- and l-RA was achieved by a separation factor of 2.08. Methylphenidate, the precursor of RA, was eluted with the front solvent, and thus does not interfere in the analysis of RA in our method. The assay was successfully applied for the in vitro analysis of enantiomeric ritalinic acids produced by human and dog plasma and dog liver.
CONCLUSIONS
Data demonstrated that the esterase(s) in human plasma metabolize d-methylphenidate faster than its l-isomer. The yielded intrinsic clearances (Clint) are 1.02 and 2.17 microl/min/mg protein, respectively, for d and l-methylphenidate.
目的
本研究旨在开发并验证一种用于定量分析人血浆中对映体形式的利他林酸（RA）的新型手性高效液相色谱 - 紫外检测法。
方法
使用α1 - 酸性糖蛋白柱，流动相含有0.4%乙酸和0.1%二甲基辛胺，pH值为3.4。对映体形式的RA在220 nm处进行检测。
结果
d - RA和l - RA实现了基线分离，分离因子为2.08。RA的前体哌醋甲酯与前沿溶剂一起洗脱，因此在我们的方法中不干扰RA的分析。该测定法成功应用于人血浆、犬血浆和犬肝脏产生的对映体形式利他林酸的体外分析。
结论
数据表明，人血浆中的酯酶代谢d - 哌醋甲酯比其l - 异构体更快。d - 哌醋甲酯和l - 哌醋甲酯产生的内在清除率（Clint）分别为1.02和2.17微升/分钟/毫克蛋白质。
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