酚麻氯汀胶囊中氯马斯汀的LC-MS-MS测定方法及在健康人体中的相对生物利用度研究

摘要

目的:建立人血浆中氯马斯汀的LC-MS-MS测定法,评价酚麻氯汀胶囊中氯马斯汀在健康人体的相对生物利用度.方法:采用开放、随机、双周期、两制剂、双序列单次给药的交叉试验设计.19例健康志愿者分别口服相当于富马酸氯马斯汀0.67 mg剂量的受试制剂和参比制剂.以硝苯地平为内标,采用甲基叔丁基醚为提取溶剂,用LC-MS-MS法测定血浆中氯马斯汀的质量浓度,经WinNonlin 6.0软件处理血药质量浓度数据后得药动学数据.结果:氯马斯汀的线性范围为5.09～407.20 ng·L-1,定量下限为5.09ng·L-1,绝对回收率为79.7%～80.6%,绝对基质效应为101.0%～103.6%,批内和批间精密度与准确度均符合要求.受试制剂中氯马斯汀的t1/2为(20.67±3.56)h,Cmax为(142.07±65.69)ng·L-1,Tmax为(4.21±1.23)h,AUC0-t为(2 829±1 681)ng·h·L-1;参比制剂中氯马斯汀的的t1/2为(20.83±4.94)h,Cmax为(1 46.55±60.16)ng·L-1,Tmax为(4.13±1.27)h,AUC0-t为(2 839±1 560)ng·h·L-1.以AUC0-t计算,与参比制剂比较,受试制剂中氯马斯汀的平均相对生物利用度为(101.7±23.4)%.结论:本方法灵敏、准确,适于临床药动学研究;两种制剂中的氯马斯汀具有生物等效性.%Objective: To develop an HPLC-MS-MS assay for determination of clemastine in human plasma , and estimate the bioequivalence of clemastine in paracetamol, clemastine fumarate and pseudoephedrine hydro-chloride capsule in healthy volunteers. Methods; An open, randomized, two-periods, two-treatment, two-sequence, and crossover clinical trial was performed in 19 healthy male volunteers. They were orally administrated with a single dose of clemastine fumarate 0. 67 mg. The plasma concentration of clemastine was determined by LC-MS-MS using nifedipine as an internal standard and methyl tert-butyl ether as an extraction solvent. The plasma concentration-time curves as well as pharmacokinetics of both test and reference formulations were analyzed using WinNonlin 6. 0 software. Results; Calibration curve of clemastine was linear in the range of 5. 09 -407. 20 ng·L-1 and LLOQ was 5.09 ng ? L-1.Absolute recovery and matrix effect were in the range of 79. 7% ~ 80. 6% and 101. 0% ~ 103. 6% , respectively. Accuracy and precision ( intra-run and inter-run) were conformed to the requirement. The main pharmacokinetic parameters of clemastine in the test and reference formulations were as follows; t1/2 were (20.67 ±3.56) and (20.83+4.94) h, Cmax were ( 142. 07 ± 65. 69 ) and ( 146. 55 ± 60. 16 ) ng·L-1, Tmax were (4.21 ± 1. 23) and (4.13 ± 1. 27) h, and AUC0-t, were (2 829 ±1681) and ( 2839 ±1560) ng·h·L . The relative bioavailability of clemastine in the test formulation was (101.7 ±23.4)% as estimated by AUC0-t. Conclusion: HPLC-MS-MS method is sensitive and accurate for determination of clemastine, and the two formulations of clemastine are bioequivalent.