LC-MS／MS法测定健康人体中氢溴酸加兰他敏的浓度及其制剂的生物等效性研究

目的：建立专属灵敏的LC-MS／MS法测定健康人体血浆中氢溴酸加兰他敏，并进行药动学及生物等效性研究。方法：对20名健康受试者采用随机双交叉试验设计，分别单剂量口服氢溴酸加兰他敏受试和参比制剂10mg，采用LC-MS／MS法测定受试者血浆中氢溴酸加兰他敏浓度，用DAS2．0软件计算相关药动学参数，评价两制剂的生物等效性。血浆样品经液-液萃取后，以甲醇-醋酸盐缓冲液（50：50）为流动相，LichrospherC6心柱（250mm×4．6mm，5μm）分离；样品经电喷雾离子源正离子化后，通过三重四极杆串联质谱仪，在多反应监测模式下对加兰他敏（m／z287．9→m/z 212．9）和内标盐酸克伦特罗（m／z277→m／z 202．7）的浓度进行测定。结果：氢溴酸加兰他敏受试和参比制剂的主要药动学参数为：cmax分别为（66．19±16．7）和（59．29±16．9）μg/L，Tmax分别为（0．59±0．28）和（0．81±0．56）h；t/2分别为（7．11±0．91）和（7．04±0．84）h，平均驻留时间分别为（9．80±1．35）和（9．87±1．18）h，AUC0-36分别为（453．47±132．77）和（445．62±87．79）μg·h／L，AUC0-∞分别为（469．05±140．30）和（460．49±92．20）μg·h／L，V/F分别为（180．10±77．71）和（201．10±74．95）L，CL／F分别为（23．99±6．20）和（24．21±5．13）L／h。以AUC0-36计算，受试制剂的相对生物利用度为（101．5±19．1）％。结论：本试验测得氢溴酸加兰他敏受试和参比制剂主要药动学参数与国内外文献报道基本一致。数据分析结果表明，两制剂生物等效。本试验方法专属性好，灵敏度高，快速简便。

Determination of the Concentration of Galanthamine Hydrobromide in Chinese Healthy Volunteers by LC-MS/MS and Study on its Bioequivalence

Objective：To establish a selective and sensitive LC-MS/MS method for the determination of the concentration of galanthamine hydrobromide in healthy human plasma and to study the pharmacokinetics and the bioequivalence of different formulations. Methods： Under the randomized cross-over design, 20 healthy volunteers were given a single dose of 10 mg galanthamine hydrobromide test or reference formulations, respectively, and plasma galanthamine concentration was determined by LC-MS/MS. The relative pharmacokinetic parameters were calculated by a DAS 2.0 program. Galanthamine hydrobromide and the internal standard clenbuterol hydrochloride were extracted from plasma by liquid-liquid extraction, then separated on a Lichrospher C6H6 column （250 mm × 4.6 mm, 5μm）. The mobile phase consisted of methanol-acetate buffer （50：50）. A Waters Qualm Micro tandem mass spectrometer equipped with electrospray ionization source was used as detector and was operated in the positive ion mode. The multiple reaction monitoring ion transitions for galanthamine and the internal standard were m/z 287.9→m/z 212.9 and m/z 277→m/z 202.7, respectively. Results：The main pharmacokinetic parmneters of galanthamine hydrobmmide in human plasma after oral 10 mg galanthamine hydrobmmide test or reference formulations were as follows： Cmax was （66.19 ± 16.7） and （59.29± 16.9） μg/L, Tmax was （0.59±0.28） and （0.81±0.56） h, tin was （7.11±0.91） and （7.04±0.84） h, mean residence time was （9.80±1.35） and （9.87±1.18） h, AUC0-36 was （453.47 ± 132.77） and （445.62 ± 87.79） μg·h/L, AUC0→∞ was （469.05±140.30） and （460.49 ± 92.20） μg·h/L, V/F was （180.10±77.71） and （201.10±74.95） L, CL/F was （23.99±6.20） and （24.21±5.13） L/h, respectively. The relative bioavailability of the test formulation was （ 101.5 ±19.1） %. Conclusion： It is suitable to evaluate bioequivalence of the test and reference formulations containing galanthamine hydrobromide, show