Validation of a Rapid and Sensitive LC-MS/MS Method for Determination of Exemestane and Its Metabolites, 17β-Hydroxyexemestane and 17β-Hydroxyexemestane-17-O-β-D-Glucuronide: Application to Human Pharmacokinetics Study
Validation of a Rapid and Sensitive LC-MS/MS Method for Determination of Exemestane and Its Metabolites, 17β-Hydroxyexemestane and 17β-Hydroxyexemestane-17-O-β-D-Glucuronide: Application to Human Pharmacokinetics Study
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DOI：
10.1371/journal.pone.0118553
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发表时间：
2015-03-20
期刊：
影响因子：
3.7
通讯作者：
Goh, Boon-Cher
中科院分区：
文献类型：
--
作者：
Wang, Ling-Zhi;Goh, Sok-Hwei;Goh, Boon-Cher
开发并验证了一种新型、快速、灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于评价人血浆中阿司美坦及其代谢产物17 β-二氢阿司美坦（活性代谢产物）和17 β-二氢阿司美坦-17-O-beta-D-葡萄糖醛酸苷（非活性代谢产物）的药代动力学。它们各自的D3同位素用作内标。使用Thermo Fisher BDS Hypersil C18分析HPLC柱（100 x 2.1 mm，5 μ m）实现分析物的色谱分离。移动的相通过用0.1%甲酸水溶液和乙腈梯度洗脱以0.5 mL/min的速率递送。色谱柱流出液采用API 4000三重四极杆质谱仪电喷雾电离（ESI）检测，多反应监测（MRM）正离子模式监测。监测到阿司美坦、阿司美坦-d3、17 β-二氢阿司美坦、17 β-二氢阿司美坦-d3、17 β-二氢阿司美坦-17-O-β-D-葡糖苷酸、和17 β-二氢甾烷-17-O-β-D-葡糖苷酸-d3。该测定方法证明，阿司美坦的线性范围为0.4-40.0 ng/mL; 17 β-二氢阿司美坦和17 β-二氢阿司美坦-17-O-β-D-葡糖苷酸的线性范围为0.2 - 15.0 ng/mL，决定系数（r（2））> 0.998。精密度（变异系数）为
A novel, rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for the evaluation of exemestane pharmacokinetics and its metabolites, 17 beta-dihydroexemestane (active metabolite) and 17 beta-dihydroexemestane-17-O-beta-D-glucuronide (inactive metabolite) in human plasma. Their respective D3 isotopes were used as internal standards. Chromatographic separation of analytes was achieved using Thermo Fisher BDS Hypersil C18 analytic HPLC column (100 x 2.1 mm, 5 mu m). The mobile phase was delivered at a rate of 0.5 mL/min by gradient elution with 0.1 % aqueous formic acid and acetonitrile. The column effluents were detected by API 4000 triple quadrupole mass spectrometer using electrospray ionisation (ESI) and monitored by multiple reaction monitoring (MRM) in positive mode. Mass transitions 297 > 121 m/z, 300 > 121 m/z, 299 > 135 m/z, 302 > 135 m/z, 475 > 281 m/z, and 478 > 284 m/z were monitored for exemestane, exemestane-d3, 17 beta-dihydroexemestane, 17 beta-dihydroexemestane-d3, 17 beta-dihydroexemestane-17-O-beta-D-glucuronide, and 17 beta-dihydroexemestane-17-O-beta-D-glucuronide-d3 respectively. The assay demonstrated linear ranges of 0.4-40.0 ng/mL, for exemestane; and 0.2 - 15.0 ng/mL, for 17 beta-dihydroexemestane and 17 beta-dihydroexemestane-17-O-beta-D-glucuronide, with coefficient of determination (r(2)) of > 0.998. The precision (coefficient of variation) were