LC-MS/MS法测定人血浆中替米沙坦的浓度及其人体生物等效性研究

目的:建立以高效液相色谱串联质谱电喷雾(LC-MS/MS)法测定人血浆中替米沙坦浓度的方法,并考察2种替米沙坦片的生物等效性。方法:人血浆样本以乙腈沉淀蛋白后,选用Zorbax SB-C_(18) Narrow Bore色谱柱,以甲醇-10mmol.L~(-1)乙酸铵(含0.5%甲酸)(80∶20)为流动相,流速为0.4mL.min~(-1);选用API3200型三重四极杆串联质谱仪的多重反应监测(MRM)扫描方式进行监测,电喷雾离子化源,正离子方式,选择监测离子反应分别为m/z515.2→276.2(替米沙坦)和m/z748.5→m/z158.2(克拉霉素,内标)。结果:替米沙坦和克拉霉素的保留时间分别为1.51、1.25min。替米沙坦血药浓度在1.00～1500ng·mL~(-1)范围内线性关系良好(r=0.9985),定量下限为1.00ng·mL~(-1);日内、日间RSD均≤6%,相对偏差(RE)均在±7%的范围以内;平均提取回收率为(93.0±3.9)%。替米沙坦片受试制剂与参比制剂平均药动学参数分别为:t_(1/2)(25.5±12.5)、(26.5±11.8)h,t_(max)(1.50±0.78)、(1.59±1.16)h,c_(max)(358±212)、(389±298)ng·mL~(-1),AUC_(0～96h)(2383±1146)、(2411±1192)ng.h.mL~(-1)。替米沙坦片受试制剂的平均生物利用度为(101.3±22.6)%。结论:该方法高效、灵敏、专属性强;2种替米沙坦片等效。

Determination of Telmisartan in Human Plasma by LC-MS/MS and Bioequivalence Study of It

OBJECTIVE：To establish LC-MS/MS method for the determination of telmisartan in human plasma and to investigate the bioequivalence of 2 kinds of Telmisartan preparations.METHODS：After protein precipitation with acetonitrile,the determination was performed on Zorbax SB-C_（18）Narrow Bore column with the mobile phase consisted of methanol-10 mmol·L^-1ammonium acetate（containing 0.5% formic acid）（80∶20）at a flow rate of 0.4 mL·min^-1.Detection was carried out by API3200 electrospray positive ionization mass spectrometry in the multiple reaction monitoring（MRM）mode.The MRM transitions of m/z 515.2→276.2 and m/z 748.5→158.2 were used to quantify telmisartan and clarithromycin（internal standard）,respectively.RESULTS：Telmisartan and clarithromycin were eluted at 1.51 min and 1.25 min,respectively.The calibration curve was linear over the concentration range of 1.00～1 500 ng·mL^-1（r=0.998 5）with the lowest detection limit of 1.00 ng·mL^-1.Intra-day RSD and inter-day RSD were both less than 6%,and the RE were within ±7%.The mean extract recovery was（93.0±3.9）%.Main pharmacokinetic parameters of test preparation and reference preparation were as follows：t1/2（25.5±12.5）h and（26.5±11.8）h,tmax（1.50±0.78）h and（1.59±1.16）h,cmax（358±212）ng·mL^-1and（389±298）ng·mL^-1,AUC0～96h（2 383±1 146）ng·h·mL^-1and（2 411±1 192）ng·h·mL^-1.The average bioavailability of telmisartan（test preparation）is（101.3±22.6）%.CONCLUSION：The method is effective,sensitive and specific,and the 2 kinds of tablets are bioequivalent.