论文部分内容阅读
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。
Objective To establish a method for simultaneous determination of irbesartan and hydrochlorothiazide in plasma by liquid chromatography-tandem mass spectrometry and to study the pharmacokinetics of human. Methods A total of 20 healthy volunteers were given a single oral test preparation (Irbesartan 150 mg, hydrochlorothiazide 12.5 mg). Blood samples were collected at different time points within 0 ~ 36 h, The internal standard was concentrated in methanol and the mobile phase was concentrated. The residue was purified on a CuroSil-PFP column (Phenomenex 250 mm × 4.6 mm, 5 μm) using 4% glacial acetic acid in water and methanol-acetonitrile : 1, V / V) gradient elution separation, electrospray ionization tandem mass spectrometry selective reaction monitoring (SRM), the positive and negative ion switching were measured irbesartan and hydrochlorothiazide in plasma concentrations. Results The irbesartan and hydrochlorothiazide plasma samples showed a good linearity with good mass spectral response at concentrations of 10 ~ 4 000 μg.L-1 and 1 ~ 400 μg.L-1, respectively. The lower limit of quantitation (LLOQ) was 10.0 μg.L-1 and 1.0 μg.L-1, respectively, with good accuracy and precision. After single oral dose of irbesartan hydrochlorothiazide tablets, the pmax, tmax, AUC0-36 and t1 / 2 of hydrochlorothiazide were (86.96 ± 34.99) μg.L-1 and (1.75 ± 0.69) h , (434 ± 143) μg.hL-1 and (7.91 ± 1.31) h respectively; the corresponding parameters of irbesartan were (1 670 ± 409) μg.L-1, (1.55 ± 0.70) h and ± 274) μg.hL-1 and (9.45 ± 5.20) h. Conclusion The method is highly specific, sensitive and accurate, which meets the requirements of irbesartan and hydrochlorothiazide pharmacokinetics studies.