LC-MS/MS法研究阿普唑仑胃漂浮片恒河猴生物等效性

Bioequiavailability study of intragastric floating tablets of alprazolam in rhesus monkeys by LC-MS/MS

目的：建立LC—MS／MS法测定恒河猴血浆中阿普唑仑的浓度，并用于阿普唑仑胃漂浮片的生物等效性分析。方法：采用随机、双周期、自身交叉试验设计，6只恒河猴单剂量口服受试制剂或参比制剂（1．2mg），不同时间点采集的血浆样本经提取．液相色谱-串联质谱法测定血浆样本中阿普唑仑的浓度，并使用DAS2．1软件计算药代动力学参数，评价制剂的生物等效性。结果：阿普唑仑的线性范围为0．4～100ng·mL^-1，定量下限为0．4ng·mL^-1，日内、日间精密度（RSD）均小于15％，准确度（RE）在．4-15％以内。恒河猴单剂量口服受试制剂和参比制剂Cmax分别为（64．75±8．78）、（78．87±6．06）ng·mL^-1，Tmax分别为（4．50±1．22）、（2．17±0．52）h，t1／2分别为（3．51±1．72）、（3．51±1．52）h，AUC0-t分别为（719．93±114．74）、（326．93±66．20）h·ng·mL^-1，相对生物利用度为（233．18±86．82）％。结论：LC—MS／MS方法适用于阿普唑仑制剂生物等效性研究。受试制剂与参比制剂比较生物利用度显著提高，Tmax显著推迟，具有缓释的效果。

Objective：To establish a method of high performance liquid chromatography tandem mass spectrometry （ HPLC - MS/MS） for the determination of alprazolam in plasma of rhesus monkeys and the bioequivalence analysis of intragastric floating alprazolam tablets. Methods： In a randomized, double - period, self - crossover study, 6 rhe- sus monkeys were given a single oral dose of test or reference tablets containing 1.2 mg alprazolam. Then the plas- ma concentration was determined by HPLC - MS/MS method after extraction of alprazolam. The pharmacokinetie parameters were calculated by DAS 2. 1 program and the bioequivalence of two formulations was evaluated. Results： The calibration curve of alprazolam was linear in the range of 0. 4 - 100 ng · mL-1. The lower limit of quantitation was 0.4 ng · mL-1. Intra - day and inter - day RSDs were both less than 15% and the relative error （RE） was within ＋ 15%. The pharmacokinetic parameters for test tablets vs reference tablets were as follows： Cmax of（64. 75 ±8.78） and（78. 87 ±6.06）ng · mL^-1 ,Tmax of（4.50 ± 1.22）and（2. 17 ±0.52）h,t1/2 of（3.51 ± 1.72） and（3.51 ± 1.52）h,AUC0-t of（719.93 ± 114.74） and（326.93±66.20）h ·ng · ml^ -1 ,the relative hioavailability was（233.18 ± 86.82）%. Conclusion： The LC - MS/MS method is suitable for the bioequivalence study of al- prazolam preparations. The statistic data showed that Tmax for test tablets was significantly delayed eomparded with reference tablets, indicating that test tablets had sustained -release effect.