HPLC-MS/MS法测定大鼠血浆中金丝桃素的浓度  

Determination of Hypericin in Rat Plasma by HPLC-MS /MS

作　　者：翟学佳[1] 陈芬[1] 吕永宁[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院药剂科,武汉430022

出　　处：《中国药师》2014年第11期1826-1829,共4页China Pharmacist

基　　金：国家自然科学基金(编号:81403012);华中科技大学自主创新研究基金青年教师基金(编号:2013QN212)

摘　　要：目的：建立HPLC-MS/MS法测定大鼠血浆中金丝桃素的浓度。方法：血浆样本加入适量内标,经乙腈直接沉淀蛋白后采用HPLC-MS/MS进行分析。色谱柱采用Ultimate C18柱（150 mm×2.1 mm,5.0μm）;流动相由乙腈-5 mmol·L^-1醋酸铵（含0.1%甲酸）（90∶10）组成,柱温35℃;流速0.5 ml·min^-1;采用电喷雾离子源（ESI）,以多反应监测方式（MRM）进行定量分析。金丝桃素和内标吡格列酮在负离子模式下定量分析离子对分别为m/z 503.2→m/z 405.1和m/z 355.0→m/z 41.9。结果：金丝桃素在0.1-13.2 ng·ml^-1线性关系良好（r〉0.99）,最低定量限为0.1 ng·ml^-1,提取回收率为84.19%-98.71%,日内、日间精密度均不高于18.47%。结论：该方法分析速度快、灵敏、准确,为临床进一步研究金丝桃素提供了基础。Objective： To establish an HPLC-MS/MS method for the determination of hypericin in rat plasma. Methods： The sample was analyzed by HPLC-MS/MS after the addition of internal standard and then protein precipitation using acetonitrile. The separation was carried out on an Ultimate C18 column （150 mm × 2. 1 mm,5. 0 μm）. The mobile phase was composed of acetonitrile∶ 5 mmol·L-1 ammoniumacetate （containing 0. 1% formic acid） （90∶10） at a flow rate of 0. 5 ml·min-1 under 35℃. The detection was performed with multiple teactions monitoring （ MRM） using an electrospray ionization （ ESI） . The precursor/product ion transitions were monitored at m/z 503. 2→m/z 405. 1 for hypericin and m/z 355. 0→m/z 41. 9 for the internal standard pioglitazone （ anion mode）. Results：The good linearity of hypericin was obtained within the range of 0. 1-13. 2 ng·ml-1. The correlation coefficient was more than 0. 99 and the lower limit of quantification was 0. 1 ng·ml-1. The extraction recovery was within the range of 84. 19%-98. 71%. The precision of intra-and inter-day was below 18. 47%. Conclusion： The method is fast, sensitive and accurate, which provides research basis for the clinical further study of hypericin.

关 键 词：金丝桃素 大鼠血浆 高效液相色谱-质谱联用法 

分 类 号：R969.1[医药卫生—药理学]