色谱

色谱 ›› 2018, Vol. 36 ›› Issue (11): 1099-1104.DOI: 10.3724/SP.J.1123.2018.07021

• 研究论文 • 上一篇    下一篇

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

贺美莲1, 郭常川2, 冷佳薇2, 张迅杰2, 咸瑞卿2, 巩丽萍2, 石峰2, 姜玮3   

  1. 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
  • 收稿日期:2018-07-23 出版日期:2018-11-08 发布日期:2014-09-30
  • 通讯作者: 姜玮.Tel:(0531)88362588,E-mail:wjiang@sdu.edu.cn;郭常川,Tel:(0531)81216561,E-mail:gcc007@163.com.
  • 基金资助:

    国家自然科学基金（81573606）.

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

HE Meilian1, GUO Changchuan2, LENG Jiawei2, ZHANG Xunjie2, XIAN Ruiqing2, GONG Liping2, SHI Feng2, JIANG Wei3   

  1. 1. School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China;
    2. Shandong Institute for Food and Drug Control, Jinan 250101, China;
    3. School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, China
  • Received:2018-07-23 Online:2018-11-08 Published:2014-09-30
  • Supported by:

    National Natural Science Foundation of China (No. 81573606).

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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50 μL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，2.5 μm），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30 mg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。

关键词: 超高效液相色谱-串联质谱, 人血浆, 生物等效性, 盐酸氨溴索

Abstract:

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 50 μL-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, 2.5 μm) 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 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.

Key words: ambroxol hydrochloride, bioequivalence, human plasma, ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)

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