高效液相色谱-串联质谱法测定人血浆中阿折地平浓度及药动学

Determination and pharmacokinetics of azelnidipine in human plasma by HPLC-MS-MS

目的:建立测定人血浆中阿折地平的HPLC-MS-MS方法,并用于阿折地平的人体药动学研究。方法:HPLC-MS-MS系统由SHIMADZU公司SIL-HTc型全自动HPLC仪和AB公司API3000型三重四极杆串联质谱仪组成,Shiseido Cap-cell Pak C18色谱柱(100mm×4.6mm,5μm),流动相:甲醇-水-甲酸(80:20:0.1),流速:0.3mL.min-1。血浆样本经乙醚液-液萃取后进样,以MRM模式检测,文拉法辛为内标。基于此检测方法进行了阿折地平单、多次给予8mg及单次给予16mg的药动学研究。结果:线性范围0.031 25～40μg.L-1,最低检测限0.031 25μg.L-1,方法回收率96.9%～102.5%,萃取回收率55.0%～63.4%,日内及日间变异系数分别为2.71%～3.36%和0.80%～2.87%。经时血药浓度数据用DAS程序进行药动学拟合,表明阿折地平的体内过程符合二室开放模型,并且有剂量依赖关系,连续给药有一定蓄积。单次口服给药8mg的tmax为(2.5±0.9)h,Cmax为(1.6±0.9)μg.L-1,AUC0-96h为(14.8±10.2)μg.L-1.h;多次口服给药8mg的tmax为(3.9±1.9)h,Cmax为(2.3±1.1)μg.L-1,AUC0-96h为(31.4±11.6)μg.L-1.h;单次口服给药16mg的tmax为(3.4±1.4)h,Cmax(5.0±3.2)μg.L-1,AUC0-96h(48.5±31.0)μg.L-1.h。结论:本测定方法灵敏、简便、特异性好,成功用于阿折地平的药动学研究取得较好的结果。

OBJECTIVE To develop an HPLC-MS -MS method for determination of azelnidipine in human plasma, and to study ist pharmaeokinetics. METHODS HPLC-MS-MS system consisted of SIL-HTc automatic HPLC instrument, Shiseido Capcell Pak C18 column （100 mm× 4. 6 mm, 5 μm）, and API 3000 triple quadrupole tandem mass spectrometer, The mobile phase was methanol-water-formic acid （80 ： 20 ： 0. 1）, the flow rate was 0. 3 mL·min^- 1. The plasma samples were extracted by liquid-liquid extraction with ether, the analyses were detected by the multiple reaction monitoring mode, venlafaxine were used as internal standard. After that, pharmacokinetic study of azelnidipine was done after single and multiple administration of 8 mg and 16 mg azelnidipine tablets. RESULTS The linear range was 0. 031 25-40μg·L^-1 , the limit of detection was 0. 031 25 μg·L^-1, the methodology recovery was 96. 9%-102. 5%, extraction recovery is 55. 0%-63. 4%, intra-and inter-assay precision is 2. 71%-3.36% and 0. 80%-2. 87%. The concentration-time data of azelnidipine were treated with DAS pharmacokinetie software and are fitted as a two-compartment open model, and there was a dose-dependent relationship, a certain accumulation of continuous administration. Singe dose administration of 8 mg;tmax was （2. 5 ± 0. 9）h, Cmax was （1.6 ± 0. 9） μg·L^-1, AUG0-96h was （ 14. 8 ± 10. 2 ） μg·L^-1· h; multiple dose administration of 8 mg ; tmax was （3. 9 ± 1.9） h, Cmax was （ 2. 3 ± 1. 1 ） μg·L^-1, AUC0-96hwas （31.4± 11.6） μg·L^-1; singe dose administration of 16 mg： tmax was （3. 4 ±1. 4）h,Cmax was （5. 0 ± 3. 2） μg·L^-1 , AUC0-96h was （48. 5 ± 31.0）μg·L^-1· h. CONCLUSION The assay is simple, rapid, and sensitive. It is success to be used on pharmacokinetics study of azelnidipine and achieves good results.