液相色谱-质谱联用法测定大鼠血浆中木通皂苷D的浓度

目的 建立液相色谱-质谱联用法(LC-MS/MS)测定大鼠血浆中木通皂苷D的浓度.方法 选用XDB-C18色谱柱,以5 mmol/L乙酸铵水溶液(含0.1％甲酸)-甲醇溶液为流动相,采用梯度洗脱进行分离.样本经甲醇沉淀后进样,选用3200QTAP型质谱仪的多重反应监测扫描方式进行检测.结果 木通皂苷D线性范围为10 ～ 1000 ng/ml,最低定量限为10ng/ml.准确度与精密度结果显示方法日间、日内变异均小于15％,相对误差为-2.8％ ～4.6％,低、中、高3个浓度提取回收率为95.3％～108.1％.结论 本研究所建立的方法快速、灵敏、专属性强、重现性好,可用于大鼠血浆中木通皂苷D浓度的测定和药代动力学研究.

LC-MS/MS determination of akebia saponin D in rat plasma

Objective To develop a HPLC-MS/MS method for the determination of aconitine and study the in vitro metabolic stability of aconitine in dog tissue homogenates.Methods The chromatographic separation was performed on a C18 column.The mobile phase consisted of acetonitrile and water with 0.2% formic acid and 5 mmol/L ammonium acetate.A triple quadrupole tandem mass spectrometer equipped with an electrospray ionization interface source was used for the quantitative determination in the positive selective reaction monitor mode.Aconitine was incubated with dog tissue homogenates and samples were withdrawn at different time points and precipitated by acetonitrile with internal standards citalopram.Results Aconitine showed good linear relationship over the range from 5 to 500 ng/ml.The recoveries of aconitine were between 85.73% and 92.12% at three QC concentration levels.The intra-and inter-day precisions were 5.32%-8.95% and 5.45%-8.86%,respectively.After incubation,about 20% of aconitine were cleared in the liver and small intestine,and t1/2 were 460.6 and 521.3 min,respectively.But none was metabolized in the stomach and kidney.Conclusion These results demonstrated that aconitine was mainly metabolized in the liver and small intestine at a slow rate.