超高效液相色谱-串联质谱法测定人血浆中盐酸氨溴索及其制剂的生物等效性评价  

Determination of ambroxol hydrochloride in human plasma by ultra high performance liquid chromatography-tandem mass spectrometry and bioequivalence evaluation of its preparation

作　　者：贺美莲 郭常川 冷佳薇 张迅杰 咸瑞卿 巩丽萍 石峰 姜玮[3] HE Meilian;GUO Changchuan;LENG Jiawei;ZHANG Xunjie;XIAN Ruiqing;GONG Liping;SHI Feng;JIANG Wei(School of Pharmaceutical Sciences,Shandong University,Jinan 250012,China;Shandong Institute for Food and Drug Control,Jinan 250101,China;School of Chemistry and Chemical Engineering,Shandong University,Jinan 250100,China)

机构地区：[1]山东大学药学院,山东济南250012 [2]山东省食品药品检验研究院,山东济南250101 [3]山东大学化学与化工学院,山东济南250100

出　　处：《色谱》2018年第11期1099-1104,共6页Chinese Journal of Chromatography

基　　金：国家自然科学基金(81573606)~~

摘　　要：采用超高效液相色谱-串联质谱(UHPLC-MS/MS)技术,建立了快速、简单、灵敏的测定人血浆中盐酸氨溴索含量的方法,并用于盐酸氨溴索人体生物等效性预试验研究。取50μL血浆样品,采用蛋白沉淀法处理,以盐酸氨溴索-d5为内标。采用Waters XBridge BEH C18色谱柱(50 mm×2. 1 mm,2. 5μm),以0. 1%(v/v)甲酸水-含0. 1%(v/v)甲酸的甲醇为流动相,在0. 4 mL/min流速下进行梯度洗脱。采用电喷雾电离(ESI)源以正离子模式进行MRM检测。结果显示,盐酸氨溴索在2～400 ng/mL范围内线性关系良好,相关系数(r)为0. 998,准确度为97. 1%～108. 7%,精密度为1. 0%～5. 6%。将该方法用于6名健康受试者口服盐酸氨溴索受试制剂和参比制剂30mg后血药浓度的测定,结果显示二者相对生物利用度为(102. 3±14. 8)%,血药浓度-时间曲线下面积(AUC0-t、AUC0-∞)和最大血药浓度(Cmax)的90%置信区间均在80. 0%～125. 0%范围内,两种制剂生物等效。A rapid,simple and sensitive ultra high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)method was developed for the determination of ambroxol hydrochloride in human plasma,and bioequivalence of its preparation was evaluated.The 50μL-plasma sample was treated with methanol for protein precipitation,while ambroxol-d5 was used as an internal standard(IS).The separation was carried out on a Waters XBridge BEH C18 column(50 mm×2.1 mm,2.5μm)by gradient elution at a flow rate of 0.4 mL/min,with 0.1%(v/v)formic acid aqueous solution and methanol containing 0.1%(v/v)formic acid as the mobile phases.The analyte was detected using an electrospray ionization source in positive ion multiple reaction monitoring(MRM)mode.The calibration curves were linear in the range of 2-400 ng/mL(r=0.998).The intra-and inter-run accuracies were 97.1%-108.7%,the intra-and inter-run precisions were 1.0%-5.6%.The method was applied to the determination of the plasma concentration of the six healthy subjects after the oral administration of 30 mg of test and reference preparations.The bioavailability was(102.3±14.8)%.The 90%confidence intervals of the test preparation’s pharmacokinetic parameters AUC 0-t,AUC 0-∞and C max were 80.0%-125.0%of the reference preparation’s corresponding parameters.Thus,it is proved that the test preparation and reference preparation are bioequivalent.

关 键 词：超高效液相色谱-串联质谱 生物等效性 盐酸氨溴索 人血浆 

分 类 号：O658[理学—分析化学]