反相高效液相色谱法测定人血浆盐酸二甲双胍浓度

Determination of metformin hydrochloride in human plasma by RP-HPLC

目的建立测定人血浆中盐酸二甲双胍浓度的反相高效液相色谱法.方法采用乙腈沉淀蛋白,苯乙双胍为内标物,高速离心后取上清液进样.流动相为0.01 mol·L-1磷酸盐缓冲液(含0.1%十二烷基硫酸钠和0.1%三乙胺,pH6.5)-乙腈(62∶ 38),流速1.0 ml·min-1,μBondapak C18柱(10 μm,3.9 mm×300 mm),柱温30 ℃,检测波长233 nm.结果盐酸二甲双胍的浓度在0.05～5.00 mg·L-1范围内线性关系良好,回归方程为Y=0.6608C+0.0009(r=0.9997),日内和日间精密度(RSD)均小于10%;检测限为0.8 ng;最低检出浓度为50 ng·ml-1. 结论本法操作简便,适用于盐酸二甲双胍血药浓度的测定及药物动力学研究.

Aim To establish a RP-HPLC method for the determination of mefformin hydrochloride in human plasma. Methods Samples of plasma were precipitated with acetonitrile,after mixing and centrifugation, the supernatant injected, The mobile phase was 0.01 mol· L^-1 KH2 PO4 （ containing 0. 1% SDS and 0. 1% triethylamine, pH6. 5 ） -acetonitrile （ 62 ： 38） , The flow rate was 1.0 ml·min^-l. Bondapak C18 column was used under 30℃. Metformin and phenoformin （ Internal standard） were detected at 233 nm. Results The of the calibration curve was liner in the range of 0.05～5.00 mg ·L ^- l （ Y = 0. 6608C ＋ 0.0009, r = 0.9997 ）, The inter-and intra-day preeisions were less than 10% （RSD＇） ;The detection limit was 0.8 ng and The limit of quantitation was 50 ng ·ml^-1. Conclusion This mothod is suitable for determination of metformin hydrochloride in human plasma and can be used in the pharmacokinetic studies.