Trontelj Jurij, Bogataj Marija, Marc Janja, Mrhar Ales
Faculty of Pharmacy, University of Ljubljana, Askerceva 7, Ljubljana, Slovenia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Aug 15;855(2):220-7. doi: 10.1016/j.jchromb.2007.05.004. Epub 2007 May 16.
This paper describes the development and validation of a method for the detection of raloxifene (Ral) and its two glucuronide metabolites, raloxifene-6-glucuronide (M1) and raloxifene-4'-glucuronide (M2), in human plasma samples. Both glucuronides were synthesized enzymatically, purified and used as authentic standards. The assay involves a simple solid phase extraction (SPE) procedure of 0.5 mL of human plasma and subsequent analysis by LC-MS-MS. The recoveries were higher than 71% and chromatographic separation of all the analytes was accomplished in less than 7 min. Linear ranges (r(2)>0.99) were found from 0.200 to 340 microg/L, from 1.600 to 2720 microg/L and from 0.088 to 60.00 microg/L, for M1, M2 and Ral, respectively. The limits of detection achieved were 8, 11 and 6 ng/L for M1, M2 and Ral, respectively. The method presented was successfully applied to a genetic polymorphism study of 47 plasma samples from women taking Evista (raloxifene hydrochloride).
本文描述了一种检测人血浆样本中雷洛昔芬（Ral）及其两种葡萄糖醛酸代谢物——雷洛昔芬 - 6 - 葡萄糖醛酸苷（M1）和雷洛昔芬 - 4'- 葡萄糖醛酸苷（M2）的方法的开发与验证。两种葡萄糖醛酸苷均通过酶法合成、纯化并用作标准品。该测定方法包括对0.5 mL人血浆进行简单的固相萃取（SPE）操作，随后通过液相色谱 - 串联质谱（LC - MS - MS）进行分析。回收率高于71%，所有分析物在不到7分钟内实现色谱分离。M1、M2和Ral的线性范围（r(2)>0.99）分别为0.200至340μg/L、1.600至2720μg/L和0.088至60.00μg/L。M1、M2和Ral的检测限分别为8、11和6 ng/L。所提出的方法成功应用于对47例服用易维特（盐酸雷洛昔芬）的女性血浆样本的基因多态性研究。
