HPLC法测定人血浆中盐酸二甲双胍浓度

Determination of metformin hydrochloride in human plasma by HPLC

目的:建立血浆中盐酸二甲双胍浓度测定方法。方法:血浆在酸性条件下以乙腈沉淀蛋白后,用二氯甲烷萃取纯化,以乙腈-5 mmol·L^(-1)磷酸盐缓冲液(pH 4．8)(45:55,每100 mL流动相中含十二烷基硫酸钠0．17 g)为流动相,色谱柱为DIAMONSIL^(TM)C_(18)柱(250 mm×4．6 mm,5μm),检测波长为233 nm,柱温为40℃。流速为1．2 mL·min^(-1)。结果:在此色谱条件下,盐酸二甲双胍与血浆中其他成分分离完全,线性范围为60．06～4 004 ng·mL^(-1),最低检测浓度为60．06 ng·mL^(-1),相对回收率在101．3％～108．3％,日内精密度RSD＜5．4％,日间精密度RSD＜13％。结论:该法稳定、灵敏、可靠,可用于人体血浆中盐酸二甲双胍浓度的测定。

Objective：To establish a HPLC procedure to determine metformin hydrochloride in human plasma. Methods：The plasma samples were deproteinized with acidified acetonitrile and purified with dichloromethane. A HPLC condition was consisted of a DIAMONSIL ODS C18 column（250 mm×4.6 mm,5μm） with the column temperature at 40 ℃, an eluate of acetonitrile-5 retool·L^-1 phosphate buffer（ pH4.8 ） （45： 55, containing SDS 0.17 g per 100 mL） with a flow rate of 1.2 mL·min^-1 and the detection wavelength at 233 nm. Results：The calibrated linear curve of metformin hydrochloride concentration was in the range of 60.06 - 4 004 ng·mL^-1. The detection limit was 60.06 ng·mL^-1. The relative recovery was 101.3%-108.3%. The within-day and among-day RSD was 〈5.4% and 〈13% , respectively. Conclusion-The simple, accurate, and reproducible HPLC method may be used for determination of plasma metformin hydrochloride.