LC-MS/MS法测定人血浆氯雷他定浓度及生物等效性研究

Determination of loratadine in human plasma by LC-MS/MS and its application in bioequivalence study

目的:建立测定人血浆中氯雷他定浓度的LC-MS/MS方法,并用于氯雷他定片人体生物等效性评价。方法:22名健康男性受试者随机交叉单剂量口服氯雷他定片受试制剂和参比制剂10 mg,采用LC-MS/MS法测定血浆样本中氯雷他定浓度。血浆样本经液-液萃取后,选用C18色谱柱,电喷雾离子化源,多重反应监测(MRM)扫描方式进行测定。用DAS软件计算其药动学参数,并评价两种制剂的生物等效性。结果:测定人血浆中氯雷他定浓度的线性范围为0.030 0～40.0 ng.mL-1(r>0.99),定量下限为0.030 0 ng.mL-1;平均提取回收率为(93.8±6.5)%;稳定性试验中,在各种贮存条件下氯雷他定均较稳定。受试制剂和参比制剂的Tmax分别为(0.84±0.36)和(0.88±0.34)h,Cmax分别为(7.82±8.10)和(8.61±11.25)ng.mL-1,t1/2分别为(7.93±6.19)和(7.31±5.13)h,AUC0～t分别为(20.3±24.9)和(19.2±24.4)ng.h.mL-1,AUC0～∞分别为(21.2±26.6)和(19.6±24.8)ng.h.mL-1。受试制剂的相对生物利用度F0～t和F0～∞分别为(100.5±34.1)%和(101.7±34.8)%。结论:该方法快速、灵敏、准确、专属性强、重现性好,适用于人血浆氯雷他定浓度的测定,并成功应用于氯雷他定片的人体生物等效性研究,两种制剂具有生物等效性。

Objective： To develop an LC-MS/MS method for determination of loratadine in human plasma, and investigate the bioequivalence of loratadine tablets in healthy volunteers. Methods： Totally 22 healthy male volunteers received a single oral close of 10 mg loratadine test and reference tablets in a randomized, two-way cross- over study. Loratadine concentrations in plasma were determined by LC-MS/MS. After liquid-liquid extraction, the analyte and the internal standard （IS） diazepam were separated on a Cxs analytical column. Detection was carried out by eleetrospray positive ionization mass spectrometry in the multiple reaction monitoring （MRM） mode. With use of DAS software, the pharmacokinetic parameters were calculated and the bioequivalence of the two preparations was evaluated. Results： The calibration curve was linear over the concentration ranges of 0. 030 0 - 40.0 ng. mL-1 with the lower limit of quantitation （LLOQ） 0. 030 0 ng. mL- 1 The mean extract recovery was （93.8 ± 6.5 ） %. In the stability studies, loratadine was found to be stable under various storage conditions. The pharmaeokinetic pa- rameters of test and reference tablets were as follows ： Tmax （0.84 ± 0.36） and （0.88 ± 0.34） h, Cmax （7.82± 8.10） and （8.61 ±11.25） ng.mL-1, t1/2（7.93 ±6.19） and （7.31 ±5.13） h, AUC0-t（20.3 ±24.9） and（19.2±24.4） ng.h.mL-1, andAUC0-∞（21.2±26.6） and （19.6±24.8） ng.h.mL-1, respectively. TheF0-t and F0-∞ of test tablet were （ 100.5 ± 34.1 ） % and （ 101.7 ± 34.8 ） % , respectively. Conclusion ： The method is rapid, sensitive, selective and reliable for the determination of loratadine in human plasma. The method has been successfully applied in a bioequivalenee study of loratadine tablets in healthy volunteers. The two preparations are bioequivalent.