液相色谱-串联质谱法同时测定人血浆中厄贝沙坦和氢氯噻嗪的浓度及药动学

目的建立液相色谱-串联质谱法同时测定血浆中厄贝沙坦和氢氯噻嗪的浓度,并用于人体药动学研究。方法 20名健康受试者单剂量口服试验制剂1片(厄贝沙坦150 mg,氢氯噻嗪12.5 mg)后,在0～36 h内不同时间点分别采集血样,分取血浆,以氯沙坦为内标,经甲醇沉淀蛋白浓缩后流动相溶解,采用CuroSil-PFP色谱柱(Phenomenex 250 mm×4.6 mm,5μm),用含4%冰醋酸的水和含4%冰醋酸的甲醇-乙腈(1∶1,V/V)的流动相梯度洗脱分离,电喷雾离子化串联质谱选择性反应监测(SRM),分别采用正和负离子切换测定厄贝沙坦和氢氯噻嗪在血浆中的浓度。结果厄贝沙坦和氢氯噻嗪血浆样品分别在10～4 000μg.L-1和1～400μg.L-1的浓度范围内质谱响应线性良好。定量下限(LLOQ)分别为10.0μg.L-1和1.0μg.L-1,准确度和精密度良好。受试者单剂量口服厄贝沙坦氢氯噻嗪片1片后,测得氢氯噻嗪的ρmax、tmax、AUC0-36和t1/2分别为(86.96±34.99)μg.L-1、(1.75±0.69)h、(434±143)μg.h.L-1和(7.91±1.31)h;厄贝沙坦的相应参数分别为(1 670±409)μg.L-1、(1.55±0.70)h、(7 396±274)μg.h.L-1和(9.45±5.20)h。结论本方法专属性强、灵敏度高、准确性高,满足厄贝沙坦和氢氯噻嗪药动学研究的要求。

Simultaneous determination and pharmacokinetics of irbesartan and hydrochlorothiazide in human plasma by LC-MS/MS

AIM To establish a LC-MS/MS method for the simultaneous determination of irbesartan and hydrochlorothiazide in human plasma,and to study the pharmacokinetics of the test compound irbesartan and hydrochlorothiazide tablets in 20 healthy volunteers after single oral dose administration.METHODS Serial blood samples were collected in heparinized tubes at given time points during 0-36 h after the drug administration(150 mg irbesartan and 12.5 mg hydrochlorothiazide).The plasma samples were treated with methanol for protein precipitation and the supernatants were separated with centrifugation and evaporated to dryness under vacuum.Losartan was used as the internal standard.The residue was reconstituted with mobile phase,and the separation was performed on a Phenomenex CuroSil-PFP(250 mm × 4.6 mm,5 μm)column with a mobile phase consisting of 4% glacial acetic acid water solution and methanol-acetonitrile(1 ∶ 1,V/V)in a gradient elution.Irbesartan and hydrochlorothiazide were determined by electrospray ionization tandem mass spectrometry in the positive and negative ion mode using selective reaction monitoring(SRM),respectively.RESULTS The calibration curves of irbesartan and hydrochlorothiazide in human plasma were linear over the concentration range from 10 to 4 000 μg·L-1 and 1.0 to 400 μg·L-1,and the low limit of quantification were 10 and 1.0 μg·L-1,respectively.The accuracy and precision were good.The plasma pharmacokinetic parameters of ρmax,tmax,AUC0-36 and t1/2 of hydrochlorothiazide were(86.96 ± 34.99)μg·L-1,(1.75 ± 0.69)h,(434 ± 143)μg·h·L-1 and(7.91 ± 1.31)h,respectively,and the corresponding parameters of irbesartan were(1 670 ± 409)μg·L-1,(1.55 ± 0.70)h,(7 396 ± 274)μg·h·L-1 and(9.45 ± 5.20)h,respectively.CONCLUSION The established LC-MS/MS method is sensitive and selective for the simultaneous determination of irbesartan and hydrochlorothiazide in plasma.It is successfully applied to a clinical pharmacokinetic study of a compound irbesartan and hydrochlorothiazide tablet.