液相色谱-串联质谱法测定人血浆中二甲双胍浓度

Determination of metformin in human plasma by HPLC-MS/MS

目的建立一种人血浆中二甲双胍浓度测定的液相色谱-串联质谱法。方法用蛋白沉淀法处理样品,色谱柱:Atlantis~?HILIC Silica(2. 1 mm×50. 0mm,5μm),流动相:10 mmol·L^(-1)醋酸铵溶液含0. 1%甲酸-90%乙腈含10mmol·L^(-1)醋酸铵和1%甲酸,流速:0. 60 m L·min^(-1),柱温:40℃,进样量:10. 0μL。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、基质效应及稳定性。结果二甲双胍在2. 00～2000. 00 ng·m L^(-1)内,线性关系良好,标准曲线为y=9. 36×10^(-3)x+2. 74×10^(-3)(r=0. 999 0),定量下限为2. 00 ng·m L^(-1),日内RSD均小于3. 4%,日间RSD均小于2. 8%,提取回收率在87. 1%以上。结论本方法简便快速,准确度高,灵敏度好,适用于人血浆中二甲双胍的浓度测定。

Objective To establish a HPLC -MS/MS method to deter- mine the concentration of metformin in human plasma. Methods The samples were treated by protein precipitation method. The chromato- graphic column was Atlantis Hilic silica （2. 1 mm×50.0 mm,5 μm） , mobile phase was 0. 1% formic acid -90% acetonitrile contained with 10 mmol·L^-1 ammonium acetate and 1% formic acid, flow rate was 0.60 mL ·min^-1, column temperature was 40℃ , injection volume was 10. 0 μL, mobile phase. The specificity, standard curve and lower limit of quantitation, precision and recovery, matrix effect and stability of the method were investigated. Results The standard curve of metformin was y =9.36 x 10 -3x ＋2. 74 ×10-3（r =0. 999 0） , and metfbrmin had good linear relationship in 2.00 - 2000.00 ng· mL^-1. The limit of quantification was 2.00 ng·mL^-1 The within - day RSD and between - day RSD were less than 3.4% and 2. 8% , respectively. The recovery rate of extraction was over 87. 1%. Conclusion The method is simple, rapid, accurate and sensitive, which is suitable for the determination of metformin in human plasma.