LC-MS/MS法测定大鼠血浆中芍药内酯苷的浓度及其在毒代动力学研究中的应用

LC-MS/MS method developed for determination of albiflorin in rat plasma and its application on toxicokinetic study

目的建立测定大鼠血浆中芍药内酯苷的液相色谱-串联质谱(LC-MS/MS)法。方法血浆样品经乙酸乙酯-异丙醇(95∶5,V/V)提取后,以Poroshell 120 EC-C18柱(50 mm×2.1 mm,2.7μm)为分析柱,乙腈-水为流动相梯度洗脱,采用ESI源在多反应监测(MRM)方式下进行正离子检测。用于定量分析的离子反应为m/z 498.5→m/z 197.1(芍药内酯苷)和m/z 251.3→m/z 108.2(拉科酰胺,内标)。结果芍药内酯苷血浆浓度测定方法的线性范围为20~2000 ng/ml,日内、日间精密度RSD%均<15%,准确度(RE)在±8.06%之间。结论该法适用于芍药内酯苷在大鼠体内的毒代动力学研究。

Objective To develop and validate a LC-MS/MS method for the determination of albiflorin in rat plasma. Meth？ods Plasma samples were extracted by liquid-liquid extraction with a mixture of ethyl acetate-isopropanol（95：5,V/V）to prepare samples for analysis. Chromatographic separation was performed on a Poroshell 120 EC-C18 column（50 mm × 2.1 mm I.D. 2.7μm）. The mobile phase consisted of acetonitrile-water followed gradient elution. Detection of albiflorin and the internal standard（IS）lacos？amide was achieved by ESI MS/MS in the positive ion mode using m/z 498.5→m/z 197.1 and m/z 251.3→m/z 108.2 transitions,respec？tively. Results The method was linear in the range of 20 to 2000 ng/ml when 50μl plasma was analyzed. The lower limit of quantifi？cation was 20 ng/ml. The inter-and intra-day precision values were both below 15%,and the accuracy（relative error）was within ± 8.06%in all quality control samples. Conclusion The method provides a sensitive and rapid means for the determination of albiflorin in rat plasma,and completely meets the requirements for toxicokinetic study.