College of Pharmacy, Catholic University of Daegu, Kyoungbuk, South Korea.
J Pharm Biomed Anal. 2010 Jan 5;51(1):264-7. doi: 10.1016/j.jpba.2009.07.023. Epub 2009 Jul 25.
Although racemic sibutramine has been widely used for the treatment of obesity, its enantioselective detection method has not been elucidated in human plasma. In this report we introduce a validated analytical method for the determination of sibutramine and its two active metabolites, desmethylsibutramines using LC-MS/MS. R- and S-isomers of those compounds in human plasma were extracted using diethyl ether-hexane (4:1, v/v) followed by an addition of NaOH solution. After removing the organic layer, the residue was reconstituted in the mobile phase 10mM ammonium acetate solution adjusted to pH 4.0 with acetic acid-acetonitrile (94:6, v/v). Both isomers in the extract were separated on a Chiralcel AGP chiral stationary-phase column and were quantified in a tandem mass spectrometry. The assay method was in accordance with FDA regulations for the validation of bioanalytical methods. This method was successfully used to profile the plasma concentrations of the stereoisomers of sibutramine and its two active metabolites with time in healthy volunteers.
尽管外消旋西布曲明已被广泛用于治疗肥胖症，但在人血浆中尚未阐明其对映选择性检测方法。在本报告中，我们介绍了一种使用 LC-MS/MS 测定西布曲明及其两种活性代谢物去甲西布曲明的验证分析方法。使用二乙醚-己烷（4：1，v/v）提取人血浆中西布曲明和其两种化合物的 R-和 S-异构体，然后加入 NaOH 溶液。除去有机层后，残留物在流动相 10mM 乙酸铵溶液中重新溶解，并用乙酸-乙腈（94：6，v/v）调节 pH 值至 4.0。在 Chiralcel AGP 手性固定相柱上分离提取物中的两种异构体，并在串联质谱中定量。该测定方法符合 FDA 对生物分析方法验证的规定。该方法成功地用于在健康志愿者中随时间描绘西布曲明及其两种活性代谢物的立体异构体的血浆浓度特征。
