液相色谱-串联质谱法测定人血浆中雷洛昔芬及其在制剂生物等效性研究中的应用  

Determination of raloxifene in human plasma using liquid chromatography tandem mass spectrometry and its application in bioequivalence study

作　　者：胡雯娟[1,2] 张逸凡[2] 陈笑艳[2] 郭丽霞[2] 夏雨[2] 钟大放[1,2] 

机构地区：[1]苏州大学,苏州215006 [2]中国科学院上海药物研究所,上海201203

出　　处：《中国新药杂志》2011年第22期2233-2237,共5页Chinese Journal of New Drugs

摘　　要：目的:建立专属、灵敏的液相色谱-串联质谱法(LC-MS/MS)法测定人血浆中的雷洛昔芬,并将方法应用于雷洛昔芬两种制剂的人体生物等效性研究。方法:血浆样品中加入200μLβ-葡萄糖苷酸酶于37℃水浴孵化10 h后采用液-液萃取法预处理。Agilent Zorbax SB-C18色谱柱(150 mm×4.6 mm,5μm)进行分离,流动相为甲醇-醋酸铵(5 mmol.L-1)-甲酸(65∶35∶0.1),流速为0.6 mL.min-1。大气压化学电离源,多反应监测方式(MRM)进行正离子检测,定量分析离子对为m/z 474→m/z 112(雷洛昔芬)和m/z 478→m/z116(内标d4-雷洛昔芬)。临床试验采用随机双交叉设计,24例健康男性受试者空腹单次口服60 mg雷洛昔芬受试制剂或参比制剂,LC-MS/MS法测定血浆雷洛昔芬浓度,计算有关药代动力学参数并进行生物等效性评价。结果:雷洛昔芬定量方法线性范围为0.20～250 ng.mL-1,定量下限为0.20 ng.mL-1,日内、日间精密度(RSD)均小于11.2%,准确度(RE)在-4.0%～1.3%之间。两种制剂的AUC0～120无显著性差异,Cmax和Tmax有显著性差异。结论:本方法专属、灵敏,适用于雷洛昔芬两种制剂的人体生物等效性评价。两种雷洛昔芬制剂吸收程度相似,但受试制剂Cmax显著降低,Tmax延长,未体现出其分散片的特点。Objective： To develop a specific and sensitive liquid chromatography-tandem mass spectrometric（LC-MS/MS） method for determining raloxifene in human plasma,and to apply this method in a bioequivalence study of two raloxifene formulations.Methods： Plasma samples were incubated with 200 μL β-glucuronidsae in water at 37 ℃ for 10 h,then 3 mL n-hexane：dichloromethane：isopropanol（20∶10∶1） was added to perform liquid-liquid extraction.Agilent Zorbax SB-C18 column（150 mm×4.6 mm,5 μm） was used as the analytical column;the mobile phase consisted of methnol： 5 mmol·L-1 ammonium acetate solution：formic acid=65∶35∶0.1 at the flow rate of 0.6 mL·min-1.The Agilent 6460 triple quadrupole mass spectrometer was operated in multiple reaction monitoring mode（MRM） via positive atmospheric pressure chemical ionization（APCI） interface using the transitions m/z 474→m/z 112 for raloxifene,and m/z 478→m/z 116 for the internal standard.In a randomized crossover study,24 healthy Chinese male subjects received a single oral administration of 60 mg either test or reference raloxifene tablets after an overnight fast.Raloxifene concentration in plasma was determined using the validated LC-MS/MS method.Results： The linear range of raloxifene in human plasma was 0.20~250 ng·mL-1 and the lower limit of quantification（LLOQ） was 0.20 ng·mL-1.Inter-day and intra-day precision（RSD） was below 11.2%.The accuracy（RE） was between-4.0% and 1.3%.There was no significant difference in AUC0~120 between two formulation,but there was significantly different in Cmax and Tmax.Conclusion： This method is specific and sensitive,and can be successfully used for the bioequivalence study of two raloxifene formulations in healthy volunteers.The absorption of two raloxifene formulations was similar.The obviously reduced Cmax and increased Tmax of test formulation did not exhibit its dispersible tablet characteristics.

关 键 词：雷洛昔芬 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] R977.12[医药卫生—药学]