RPHPLC法测定人血浆中二甲双胍浓度

An improved RPHPLC method for the determination of metformin in human plasma

目的:建立测定人血浆中二甲双胍浓度的反相高效液相色谱法。方法:人血浆标本以盐酸阿替洛尔为内标,酸化后以乙腈沉淀蛋白、二氯甲烷反洗上清液后,取上清液进行色谱分析,以5 mmol/L SDS∶0.04 mol/L KH2PO4∶CH3OH=1∶1∶1.7为流动相,流速1 ml/min,经Agilent Zorbax SB C18(150×4.6 mm,5μm)柱分离,检测波长240 nm。结果:二甲双胍标准曲线范围10～5 000μg/L,y=0.001 12x+0.012 671(r=0.999 4),最低检测限为10μg/L,高、中、低3种浓度的日内精密度均<7%,日间精密度均<8%,萃取回收率75.6%～86.6%。结论:该方法简便、快速、准确,可用于人体内二甲双胍药动学研究和临床血药浓度监测。

Objective： To describe and validate a RPHPLC-UV method for the assay of mefformin in human plasma. Methods： Atenolol was used as internal standard. Acidified samples of plasma were deproteinated with acetonitrile, washed with dichloromethane and the resulting supernatant was injected for chromatographic separation. The condition of RPHPLC was a Agilent Zorbax SB C 18 column （150×4.6 mm, 5 μm）, mobile phase of 5 mmol/L SDS ： 0.04 mol/L KH2PO4： CH3OH= 1： h 1.7 with a flow-rate of 1 ml/min, and UV detector at 240 nm. Results： The calibration curve was linear（r =0.999 4） within the range of 10- 5 000 μg/L for mefformin. Limit of quantization for MET were 10 μg/L. Daily and daytime RSD were less than 7% and 8% respectively. Mean recoveries for MET were 75.6%-86.6%. Conclusion： The method is suitable for pharmacokinetic studies and therapeutic drug monitoring.