艾司奥美拉唑UPLC-MS/MS测定方法建立及其在健康中国人中的生物等效性研究

目的 建立UPLC-MS/MS法测定人血浆中艾司奥美拉唑浓度，并应用于人体生物等效性研究。方法 采用UPLCMS/MS法，以艾司奥美拉唑-d3为内标，ACE 3 C₁₈-AR （50 mm×2.1 mm，3 μm）为分析柱，流动相A为5 mmol·L^-1乙酸铵水溶液，流动相B为乙腈，体积流量为0.6 mL·min-1进行梯度洗脱；采用电喷雾离子源，以多反应监测（MRM）正离子模式进行检测，用于定量分析的离子对为m/z 346.1→198.1（艾司奥美拉唑）、m/z 349.1→198.1（艾司奥美拉唑-d3）。应用建立的方法测定人血浆中艾司奥美拉唑浓度，并计算生物等效性。结果 艾司奥美拉唑在3~3 000 ng·mL^-1范围线性关系良好（r ≥ 0.999 4），批内、批间精密度和准确度、回收率、基质效应及稳定性均符合生物样品测定要求。应用该方法进行人体生物等效性样品测定，证明受试制剂和参比制剂生物等效。结论 建立的人血浆中艾司奥美拉唑浓度测定方法准确、灵敏、选择性好、重现性高，可用于受试者单次口服40 mg艾司奥美拉唑镁肠溶干混悬剂的生物等效性研究，受试制剂和参比制剂生物等效。

Determination of esomeprazole in human plasma by UPLC-MS/MS and its bioequivalence study in healthy Chinese subjects

Objective To established and validated a UPLC-MS/MS method for the determination of esomeprazole in human plasma, which was applied to bioequivalence study. Methods UPLC-MS/MS method using esomeprazole-d3 as the internal standard(IS) was used to determine esomeprazole in human plasma. The determination was performed on ACE 3 C₁₈-AR (50 mm×2.1 mm, 3 μm) with a linear gradient, the flow rate was 0.6 mL·min-1. The mobile phase A consisted of water containing 5 mmol·L^-1 ammonium acetate, the mobile phase B consisted of acetonitrile. The ion transitions under multiple reaction monitoring (MRM) mode by ESI⁺ ionization were performed at m/z 346.1→198.1 and m/z 349.1→198.1 for carbamazepin and esomeprazole-d3. The established method was used to determine the concentration of esomeprazole in human plasma in the bioequivalence study. Results The liner range of esomeprazole were 3-3 000 ng·mL^-1(r ≥ 0.999 4). The intra-day and inter-day accuracies and precisions, the recoveries and matrix effects, and the sample stability were all suitable to the biological samples determination. The method was applied to determine human plasma samples of bioequivalence study, and it was proved that the test preparation and the reference preparation were bioequivalent. Conclusion The method was proved to be highly accurate, sensitive, selective, and suitable for the bioequivalence study of esomeprazole in healthy Chinese volunteers after a single oral dose of 40 mg esomeprazole suspension. The test preparation and the reference preparation were bioequivalent.