S. K. Patel College of Pharmaceutical Education and Research, Ganpat University, Gujarat, India.
Acta Pharm. 2012 Mar;62(1):45-58. doi: 10.2478/v10007-012-0004-3.
A simple RP-HPLC method for the quantification of valsartan (VAL), amlodipine (AML) and hydrochlorothiazide (HCT) in human plasma was developed and validated. VAL, AML and HCT were resolved using a Gemini C18 column and mobile phase gradient starting from 20 % acetonitrile and 80 % 10 mmol L(-1) ammonium formate (V/V, pH 3.5 ± 0.2, by formic acid) to 70 % acetonitrile and 30 % 10 mmol L(-1) ammonium formate, over 20 minutes, with a flow rate of 1 mL min(-1). The samples were purified by protein precipitation and extraction. Telmisartan was used as internal standard. The method was validated according to USFDA and EMEA guidelines with good reproducibility and linear responses R = 0.9985 (VAL), 0.9964 (AML), and 0.9971 (HCT). RSDs of intra- and inter-day precision ranged 2.2-8.1 and 4.6-11.7 %, respectively, for all three drugs. Mean extraction recoveries of three QCs for the triple drug combination were 76.5 (VAL), 72.0 (AML) and 73.0 (HCT) % for human plasma. Although the LC-MS/MS method is more sensitive than HPLC, HPLC is still suitable for preliminary pharmacokinetic study. The experiments performed demostrated that simultaneous determination of all components of the triple drug combination in human plasma can be done by this method. Proposed method can be also used for guidance to the LC-MS/MS method.
建立并验证了一种用于人血浆中缬沙坦（VAL）、氨氯地平（AML）和氢氯噻嗪（HCT）定量的简单反相高效液相色谱（RP-HPLC）方法。VAL、AML 和 HCT 采用 Gemini C18 柱和流动相梯度洗脱分离，起始条件为 20%乙腈和 80%10mmol/L 甲酸铵（V/V，pH3.5±0.2，用甲酸调），20 分钟内逐渐转变为 70%乙腈和 30%10mmol/L 甲酸铵，流速为 1mL/min。样品经蛋白沉淀和提取进行净化。替米沙坦用作内标。方法按照美国 FDA 和欧洲药品管理局（EMEA）指南进行验证，具有良好的重现性和线性响应（R=0.9985（VAL）、0.9964（AML）和 0.9971（HCT））。三种药物的日内和日间精密度的 RSD 分别为 2.2-8.1%和 4.6-11.7%。三种 QC 的平均提取回收率为 76.5%（VAL）、72.0%（AML）和 73.0%（HCT），用于人血浆中的三药组合。尽管 LC-MS/MS 方法比 HPLC 更灵敏，但 HPLC 仍适用于初步药代动力学研究。实验表明，该方法可用于同时测定人血浆中三药组合的所有成分。该方法也可用于指导 LC-MS/MS 方法。
