RP-HPLC法测定人血浆中氯沙坦钾、厄贝沙坦和格列齐特浓度

Simultaneous determination of losartan,irbesartan and gliclazide in human plasma by RP-HPLC

目的建立人血浆中氯沙坦钾、厄贝沙坦和格列齐特的反相高效液相色谱(RP-HPLC)定量分析方法。方法血浆样品中加入内标物(苯妥英钠),在酸性条件下用乙酸乙酯提取,氮气吹干重溶后进行RP-HPLC分析;采用Hypersil ODS-C_(18)色谱柱(200mm×4.6 mm,5μm);流动相为乙腈-0.02 mol·L^(-1)NaH_2PO_4缓冲液(用磷酸调pH=2.98)梯度洗脱;流速1.0 mL·min^(-1);紫外检测波长228 nm。结果在选定的色谱条件下,氯沙坦钾、厄贝沙坦、格列齐特和苯妥英钠保留时间分别约为8.797、11.350、14.084、5.525 min。血浆样品中氯沙坦钾线性范围为0.05~0.6 mg·L^(-1)(r=0.999 7),最低检测浓度为0.0lmg·L^(-1);厄贝沙坦线性范围为O.1~8.O mg·L^(-1)(r=0.9999),最低检测浓度为0.03 mg·L^(-1);格列齐特线性范围为O.1~12.0mg·L^(-1)(r=0.999 9),最低检测浓度为0.03mg·L^(-1)。三者提取回收率均>80%,日内、日间RSD均<10%。结论该方法灵敏简便、准确、重复性较好,可用于氯沙坦钾、厄贝沙坦及格列齐特的血药浓度测定。

AIM To establish a RP-HPLC quantitative analysis method for the detennination of losartan, irbesar- tan and gliclazide in human plasma. METHODS The sample was treated by adding phenytoin sodium as an internal standard. The plasma was extracted with ethyl acetate in acidic condition. The solvent was evaporated to dryness by N2. The residue was dissolved and analyzed by RP-HPLC. The RP-HPLC was conducted on a Hypersil ODS-CI8 column （200 mm x 4.6 mm, 5 p_m）. The mobile phase was acetonitrile and 0.02 mol＂ L- 1 NaHEPO4 buffer solution （ adjusted pH to 2.98 with phosphoric acid） in the gradient elution. The flow rate was 1.0 mL. min-1, and the detection wavelength was 228 rim. RESULTS Using this method, the retention time of losartan,irbesartan, gliclazide, and phenytoin sodium was about 8.797,11.350,14.084 and 5.525 min, respectively. The linear ranges of losartan were 0.05 -0.6 mg.L-1 （ r = 0. 999 7）, the limit of detection was0. 01 mg- L- 1. The linear ranges of irbesartan were 0. 1 - 8.0 mg- L- 1 （ r = 0.999 9）, the limit of detection was 0. 03 mg- L- 1. The linear ranges of gliclazide were 0. 1 - 12.0 mg- L- 1 （ r = 0 999 9）, the limit of detection was0. 03 mg. L- 1. The extraction recovery rate was more than 80 %, and the relative standard deviation of intra-day and inter-day was less than 10%, respectively. CONCLUSION The proposed method is simple, accurate and specific for the determination of losan, irbesartan and gliclazide.