用HPLC—MS／MS法测定人血浆中的二甲双胍浓度

目的：建立高效液相色谱-质谱／质谱（HPLCMS／MS）联用法测定人血浆中二甲双胍浓度的方法。方法：血浆样品经乙腈沉淀蛋白质后，用HPLC—MS／MS法测定二甲双胍浓度。甲醇-乙腈10mmol／L乙酸铵溶液（20：20：60）为流动相，流速为0．2mL／min；采用大气压化学电离源，以多离子反应监测（MRM）方式进行正离子检测，二甲双胍和内标苯乙双胍的定量分析离子对分别为m／z 130．2→71．0和m／z 206．1→59．9。结果：血浆样品中二甲双胍线性范围为2．0～2000μg／L（n=7，r=0．9996），最低定量限为2．0μg／L。低、中、高（10、500、1500 μg／L）3种浓度的日内RSD分别为4．65％、2．84％和3．41％（n=5）；日间RSD分别为7．96％、3．98％和4．77%（n=5）；准确度为95．9％、97．4％和89．90%（n=5）。结论：本方法灵敏度高、专属性强、重现性好、准确，可用于人血浆中二甲双胍的含量测定。

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

Objective： To establish a HPLC-MS/MS method for determining the concentration of metformin in human plasma. Methods： Metformin and phenformin （internal standard） were separated from plasma by protein precipitation with acetonitrile, and determined by HPLC-MS/MS method with the mobile phase consisting of methanol acetonitrile-10 mmol/L ammonium acetate （20 ： 20 ： 60）, at a flow rate of 0.2 mL/min. Atmospheric pressure chemical ionization（APCI）source was applied and operated in the positive ion mode in multiple reaction monitoring （MRM） mode. The fragmentation transitions from precursor to production were m/z 130.2→71.0 for metformin and m/z 206. 1→59.9 for phenformin,respectively. Results： The linear range of metformin was 2.0 -2 000 μg/L（n=7,r =0. 999 6）. The lowest limit of quantitation of metformin was 2.0 μg/ L. At low, middle and high concentrations （ 10,500,1 500 μg/I,）, the intra-day RSDs were 4.65 %, 2.84 % and 3.41 % （n= 5）, the inter-day RSDs were 7.96%,3.98% and 4.77%（n=5）,and the accuracy rates were 95.9% ,97.4% and 89.9%（n=5）. Conclusion： The method is sensitive,specific,reproducible and accurate for the determination of metformin in human plasma.