LC-MS/MS法测定血浆中米屈肼浓度及其人体药代动力学研究

LC-MS/MS determination of plasma mildronate and its pharmacokinetics in human

目的:建立以高效液相色谱串联质谱(LC-MS/MS)检测法测定人血浆中米屈肼浓度的方法,并研究其在健康人体内的药代动力学。方法:以对乙酰氨基酚为内标,血浆样品经甲醇蛋白沉淀后,采用DIK-MA Inspire C18(150 mm×4.6 mm,5μm)色谱柱进行分离,流动相为甲醇-10 mmol.L-1醋酸铵水溶液(55∶45,v/v),质谱采用ESI源,正离子检测模式,以选择性反应监测方式进行测定,用于定量分析的离子反应分别为:m/z 147.2→58.3和m/z 152.0→110.0。结果:米屈肼血药浓度在0.01～20μg.mL-1范围内线性关系良好(r=0.997);日内、日间精密度均≤7.6%;提取回收率(低、中、高)分别为(83.6±5.1)%,(89.7±5.0)%和(89.3±3.6%)%;介质效应(低、中、高)分别为(95.3±4.5)%,(93.6±2.4)%和(98.0±2.0)%。结论:本方法适用于米屈肼的人体药代动力学研究。

Objective： To establish an LC-MS/MS method for the determination of mildronate in human plasma and to study its pharmacokinetics in healthy subjects.Methods： Acetaminophen was used as internal standard.After protein precipitation of plasma simples with methanol,the analytes were separated on a DIKMA Inspire C18 column（150 mm×4.6 mm,5 μm） with methanol and 10 mmol·L-1 ammonium acetate（55∶45,v/v）,and then analyzed by mass spectrometry with ESI ion source in the positive ion multiple reaction monitoring（MRM） mode.The target compounds mildronate and acetaminophen were quantified with m/z 147.2→58.3 and m/z 152.0→110.0.Results： The linear range of mildronate was 0.01~20 μg·mL-1（r=0.997）.The inter-day and intra-day precisions were less than 7.6 %.The extraction recoveries of mildronate at low,middle and high levels were（83.6±5.1）%,（89.7±5.0）% and（89.3±3.6）%,respectively.Matrix effects of mildronate at low,middle and high levels were（95.3±4.5）%,（93.6±2.4）% and（98.0±2.0）%,respectively.Conclusion： The method developed in this study is suitable for the pharmacokinetic study of mildronate.