LC-MS-MS法测定人血浆中米格列奈浓度

LC-MS-MS determination of mitiglinide in human plasma

目的建立液质联用色谱法(LC-MS-MS)测定人血浆中米格列奈(mitiglinide)浓度。方法空白血浆加米格列奈,用乙腈做为沉淀剂,取上清液用于LC-MS-MS分析。分析柱为日本资生堂CAPCELL PAK C18 MG5,μm150 mm×2.0 mm(ID)柱,流动相为乙腈-10 mmol/L乙酸铵(0.1%氨水)分段洗脱(0～2 min 10∶90,2.1～8 min 90∶10,8.1～10 min 10∶90),流速为0.3 ml/min;质谱条件:电喷雾离子化电离源ESI负离子检测,喷雾电压(SP)4000 kV,鞘气(SGP)流速30 Arb,辅助气(AGP)流速0 Arb,毛细管温度(TEM)314℃;米格列奈和内标瑞格列奈的碰撞能量均分别为21 EV、23 EV;选择反应监测(SRM)分别测定米格列奈和内标瑞格列奈负离子m/z 314→189和m/z451→379.2。结果米格列奈在2.5～2000μg/L检测浓度范围内呈良好线性关系(R>0.99),最低定量限(LLOQ)为2.5μg/L,绝对回收率在70%以上,高中低3种浓度的日内和日间RSD≤15%。结论该方法操作简便、灵敏、准确,适用于临床米格列奈的血药浓度监测及其药动学研究。

Objective To establish an LC-MS-MS method for determining mitiglinide in human plasma.Methods Mitiglinide added into blank plasma was sedimented by methanol,and the supernatant was determined by LC-MS-MS.The analytical column was CAPCELL PAK C18 MG,5 μm 150 mm×2.0 mm（ID）.The mobile phase consisted of acetonitrile-10 mmol/L ammonium acetate（0.1% ammonia solution）,grading elution（0-2 min 10∶90,2.1-8 min 90∶10,8.1-10 min 10∶90） at a flow rate of 0.3 ml/min.Mass spectrum conditions included ESI that was performed in the SRM mode using target ions m/z 314→189（21 EV）（mitiglinide）,m/z 451→379.2（23 EV）（repaglinide）,SP 4000 kV,SGP 30 Arb,AGP 0 Arb and TEM 314℃.Results The calibration curve was linear over the range of 2.5-2000 g/L.The LLOQ of mitiglinide in plasma was 2.5 g/L.The extracted recovery was above 70%.The intra-and inter-day RSD was below 15%.Conclusion This method is sensitive,simple and accurate to determinee mitiglinide plasma concentration and to study pharmacokinetics of mitiglinide.