柱前衍生,LC-MS/MS同时测定血浆中的孕二烯酮、依托孕烯和炔雌醇  

Simultaneous determination of gestodene,etonogestrel and ethinylestradiol in plasma by LC-MS/MS following derivatization

作　　者：刘笑芬[1] 丁存刚[1] 葛庆华[1] 周臻[1] 支晓瑾[1] 

机构地区：[1]上海医药工业研究院,药物制剂国家工程研究中心,上海201203

出　　处：《药学学报》2010年第1期87-92,共6页Acta Pharmaceutica Sinica

基　　金：国家“重大新药创制”科技重大专项子项目(2009ZX09310)

摘　　要：建立LC-MS/MS同时测定血浆中孕二烯酮、依托孕烯和炔雌醇的方法。血浆样品经液-液萃取、柱前衍生处理后,采用ESI离子源,多反应离子监测(MRM),正离子扫描进行测定。以炔诺孕酮为内标,采用C18(100mm×2.1mm,5μm)柱,梯度流动相,梯度流速测定血浆中孕二烯酮、依托孕烯和炔雌醇。结果表明,孕二烯酮、依托孕烯分别在0.1～20ng·mL-1、炔雌醇在0.01～2ng·mL-1时线性关系良好,精密度小于10.0%、方法回收率在93.6%～110.9%。对复方孕二烯酮和复方依托孕烯透皮给药制剂进行了家兔的药动学研究。本方法灵敏度高(孕二烯酮、依托孕烯达到100pg·mL-1,炔雌醇达到10pg·mL-1)、重现性好,适合药动学研究。To establish a sensitive and specific method for simultaneous determination of gestodene, etonogestrel and ethinylestradiol in plasma by LC-MS/MS, plasma samples were extracted and derivatized before injection. An ESI ion source was used and operated in the positive ion mode with multiple reaction monitoring （MRM）. Norgestrel was chosen as internal standard and performed on a C18 （100 mm × 2.1 mm, 5 μm） column. The concentrations of gestodene, etonogestrel and ethinylestradiol were measured, using step-gradient mobile phase and step-gradient flow rate. The method was validated over the concentration range of 0.1-20 ng·mL-1 for gestodene and etonogestrel and 0.01-2 ng·mL-1 for ethinylestradiol, and showed excellent linearity. The intra- and inter-assay accuracy and precision were below 10.0% and recovery was 93.6%-110.9% over the three concentration levels evaluated. The method was applied in pharmacokinetic study of the compound gestodene patch and the compound etonogestrel patch in rabbits. The LC-MS/MS method was selective, accurate and sensitive, especially the LOQ were 100 pg·mL-1 for gestodene and etonogestrel and 10 pg·mL-1 for ethinylestradiol. The method was successfully applied in pharmacokinetic study for contraceptives.

关 键 词：孕二烯酮 依托孕烯 炔雌醇 柱前衍生 LC-MS/MS 

分 类 号：R917[医药卫生—药物分析学]