液相色谱-串联质谱法测定苯妥英在健康人体的血药浓度并研究其生物等效性  

Determination the concentration of phenytoin in healthy human plasma and study on its bioequivalence by LC/MS/MS

作　　者：王丹[1] 陈笑艳[2] 张逸凡[2] 李小燕[1] 孔璋[1] 钟大放[2] 

机构地区：[1]沈阳药科大学药物代谢与药物动力学实验室,辽宁沈阳110016 [2]中国科学院上海药物研究所药物代谢研究中心,上海201203

出　　处：《中国临床药理学杂志》2006年第3期213-217,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的建立快速、灵敏的液相色谱-串联质谱法测定健康人血浆中的苯妥英(抗癫痫药),并进行生物等效性研究。方法血浆样品50μL经液- 液萃取后,以甲醇-水-甲酸(90:10:0．2)为流动相,Zorbax SB-C18柱分离; 样品经大气压化学电离源(APCI)正离子化后,通过三重四极杆串联质谱仪, 用选择反应监测(SRM)对苯妥英(m／z 253→m／z 182)和柳胺酚(m／z 230→ m／z 121,内标)进行测定。用此法测定了20名受试者单剂量口服受试和参比制剂后苯妥英的血药浓度。结果线性范围为2．5—3 000 ng·mL-1,定量下限为2．5 ng·mL-1;日内、日问精密度(RSD)均<7．0％,准确度(RE)在 -0．5％～2．3％。2种制剂的Cmax、AUC0-t均无显著性差异。结论该法专属、灵敏、快速,适用于复方制剂中苯妥英的生物等效性评价。Objective To establish a sensitive and selective LC/MS/MS method for the determination of phenytoin in healthy human plasma to study on the bioequivalence of different formulations containing phenytoin. Methods Phenytoin and the internal standard osalmid were extracted from plasma with liquid - liquid extraction, then separated on a Zorbax SB - C18 column. The mobile phase consisted of methanol - water -formic acid （90： 10： 0.2）, at a flow - rate of 0.6 mL·min^-1 A Finnigan TSQ quantum ultra tandem mass spectrometer equipped with an atmospheric pressure chemical ionization （APCI） source was used as the detector and operated in the positive ion mode. Selected reaction monitoring （SRM） using the precursor to product ion combinations of m/z 253→m/z 182 and m/z 230→m/z 121 was performed to detect phenytoin and the internal standard, respectively. The method was used to evaluate the bioequiavailability of phenytoin in compound preparation. Results The linear calibration curves were obtained in the concentration range of 2.5 -3000 ng·mL^-1 for phenytoin. The lower limit of quantification was 2.5 ng·mL^-1. The intra - and interday RSD over the entire concentration range were less than 7.0%. Accuracy determined at three concentrations （2. 5, 60.0 and 2 400. 0 ng·mL^-1 for phenytoin） ranged from - 0.5% to 2.3%. The bioavailability of two formulations was （ 107.5±18.9） %. Conclusion The method was proved to be ro -bust, convenient and suitable for bioequivalence evaluation of different formulations containing phenytoin.

关 键 词：苯妥英 液相色谱-串联质谱法 生物等效性 

分 类 号：R969.1[医药卫生—药理学] R971.6[医药卫生—药学]