一种快速灵敏测定人血清中二甲双胍含量的方法

Rapid and sensitive method for the determination of metformin in human serum

目的 建立一种快速、准确测定血清中盐酸二甲双胍含量的HPLC方法。方法 在血清中加入含有内标的乙腈溶液沉淀蛋白,离心后的上清液加二氯甲烷,萃取其中的乙腈,浓缩上清液,浓缩后的上清液直接进样分析。结果 盐酸二甲双胍在测定的2.49～0.01mg·L^(-1)浓度范围内呈现良好线形,不受杂质干扰,加权回归系数γ=0.9998。日内变异系数小于4.0％,日间变异系数小于10.65％。方法的绝对回收率在80.5％～93.3％。结论 该方法可用于盐酸二甲双胍缓释片的药代动力学研究。

Objective A quick and accurate high performance liquid chromatography (HPLC) method was developed for the determination of metformin in human serum. Methods Human serum was precipitated with acetonitrile containing internal standard, after mixing and centrifugation, the supernatant was washed with dichloromethane, after centrifugation, a 40uL of the remained supernatant was injected into an Alltech Si column monitored with UV detector at 233 nm. The mobile phase containing 0.05 mol·L-1 phosphate buffer (pH = 6.0) and acetonitrile in a 40:50 ratio pumped at a flow rate of 1.0 mL.min-1. Results The assay of metformin hydrochloride in human serum was specific and linear over the range of 2.49-0.01 mg·L-1 with a weighted coefficient (y =0.9998). In human serum the within-day RSD were less than 4.0%, and between-day RSD were less than 10.65%. The absolute recovery rates were within 80.5%~93.3%. Conclusion This assay method is rapid, accurate, simple and was used in the phamarmacokinetic study of metformin hydrochloride extended-release formulation.