液相色谱-串联质谱法测定人血浆中米格列奈浓度及其药动学研究

Determination of mitiglinide in human plasma by LC-MS/MS and study of pharmacokinetics

目的建立一种快速分析测定人血浆中米格列奈的液相色谱-串联质谱色谱法,用以研究米格列奈在健康人体内的药动学。方法以那格列胺为内标,血浆酸化后经液液萃取后,采用液相色谱-串联质谱法以多反应检测方式进行测定,选择监测的离子为m/z 316.2→298.2(米格列奈)和m/z 318.2→120.2 (那格列胺)。流动相以甲醇-10 mmol·L^(-1)醋酸铵水溶液(75:25,V/V),流速0.3 mL·min^(-1),色谱柱为Agilent Zorbax Eclipse Plus C_(18)(1.8μm,3 mm×150 mm);柱温:30℃。结果米格列奈在0.502 0～4 016μg·L^(-1)浓度范围内呈良好的线性关系(r=0.995),最低检测浓度为0.502 0μg·L^(-1),精密度和准确度试验均符合生物分析要求,应用此法测得5、10、20 mg剂量组不同给药方式、多个时间点的米格列奈血药浓度,结果呈线性动力学特征。结论该方法灵敏度高、专属性强、准确、简便,适用于米格列奈的人体药动学研究。

AIM To determine the concentration of mitiglinide in human plasma by LC-MS/MS and to study its pharmacokinetics. METHODS Acidified plasma samples were prepared by a simple liquid-liquid extraction with ethyl acetate. The chromatographic separation was performed on an Agilent Zorbax Eclipse Plus C18 column with a mobile phase of methanol -10 mmol·L ^-1 ammonium acetate solution. Analytes were detected with an Agilent 6410 Triple qudrupole mass spectrometer equipped with an electrospray ionization source in positive multiple reaction monitoringmode： m/z 316.2 （precursor ion） to 298.2 （product ion） for mitiglinide and m/z 318.2 （precursor ion） to 120.2 （product ion） for the internal standard. RESULTS The standard curve was linear （r = 0.995） over the concentration range of 0.502 0 - 4 016 μg·L^-1. Accuracy and precision were all within ± 15 %. The lower limit of quantification for mitiglinide was 0.502 0 μg·L^-1. The linear pharmacokinetics was detected in plasma concentrations of the single oral dose of 5 mg, 10 mg and 20 mg group in these healthy volunteers. CONCLUSION A rapid, sensitive and simple LC-MS/MS method for determination of mitiglinide was development, and the assay was successful applied to a clinical pharmacokinetic study.