LC-MS-MS法测定人血浆中米格列奈浓度

摘要

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 MG,5 μm 150 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/z 451→379.2.结果 米格列奈在2.5～2000 μg/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2.5 μg/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 x 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.