HPLC-MS/MS法同时测定大鼠血浆中槐角苷及染料木素

目的: 建立快速、灵敏的液相色谱-串联质谱方法测定大鼠血浆中槐角苷及其苷元染料木素。方法: 血浆经过乙酸乙酯提取,以大豆苷元为内标,采用Zorbax SB-C₁₈(2.1 mm×30 mm,3.5 μm)色谱柱,流动相为甲醇-0.001%甲酸铵溶液(加氨水调节pH至7.5)(45: 55),流速0.2 mL·min^-1;质谱条件为电喷雾离子源(ESI源),负离子模式检测,扫描方式为多反应监测(MRM),定量离子对为m/z 252.9→223.7(大豆苷元)、m/z 431.1→267.6(槐角苷)、m/z 268.8→132.8(染料木素)。结果: 槐角苷浓度在1.072～536 ng·mL^-1,染料木素浓度在1.068～534 ng·mL^-1的范围内线性关系良好(r>0.995);日间和日内精密度RSD均小于10%,低、中、高3个浓度的提取回收率在85%～97%之间。结论: 本法可用于大鼠血浆中槐角苷和染料木素的测定及其药代动力学研究。

Objective: To develop a quick and sensitive HPLC-MS/MS method for determination of sophoricoside and genistein in rat plasma. Methods: The plasma was extracted by ethyl acetate.Sophoricoside and genistein with dazein as internal standard were separated on a Zorbax SB-C₁₈ (2.1 mmm×30 mm, 3.5 μm) column with a mobile phase of methanol-0.001% formic ammonate (adjust pH to 7.5 by ammonia) (45: 55) at a rate of 0.2 mL·min^-1 and analyzed by HPLC-triple quadrupole MS with an ESI ion source in MRM mode using the transition m/z 252.9→223.7, m/z 431.1→267.6, 268.8→132.8 for dazein, sophoricoside and genistein respectively. Results: Excellent linear relationships were obtained in the range of 1.072-536 ng·mL^-1 for sophoricoside and 1.068-534 ng·mL^-1 for genistein (r>0.995).Both intra-day and inter-day RSDs were less than 10%, and the relative recoveries at three levels ranged 85%-97%. Conclusion: This method can be used to determine sophoricoside and genistein in rat plasma and its pharmacokinetic study.

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