超高效液相色谱-串联质谱法研究淫羊藿苷在大鼠体内药代动力学

UPLC-MS / MS for the pharmacokinetics of icariin in rats

目的建立超高效液相色谱-串联质谱法（UPLC-MS/MS）测定大鼠血浆中淫羊藿苷（ICA）浓度的方法,并用于研究大鼠灌胃ICA的药代动力学。方法采用乙酸乙酯提取血浆中ICA,以香豆雌酚（CMT）为内标,在BEH C18（50 mm×2.1 mm,1.7μm）色谱柱上以含0.1%（V/V）甲酸的乙腈-2 mmol/L甲酸铵的水溶液为流动相梯度洗脱,流速0.3 m L/min;以负离子电喷雾多反应监测方式定量分析ICA（m/z675.6→351.1）和CMT（m/z 267.0→211.1）。大鼠多次灌胃50 mg/kg ICA,采集不同时刻血样并测定药物浓度。结果 ICA的线性范围为0.5-50 ng/m L,方法定量下限为0.5 ng/m L。ICA日内和日间精密度（RSD）在11.3%以内,准确度在94.3%-98.7%,提取回收率为81.3%-85.2%,基质效应为94.3%-103.2%。大鼠灌胃ICA的t1/2为（1.68±0.29）h,C_（max）为（29.6±5.3）ng/m L,t_（max）为（1.00±0.00）h,AUC0-t为（88.4±13.9）（h·ng）/m L。结论 UPLC-MS/MS法专属性好、准确度高,适用于ICA临床前药代动力学研究。

Objective To establish an ultra-performance liquid chromatographytandem mass spectrometry（ UPLC-MS / MS） method for the quantification of icariin（ ICA）, and investigate pharmacokinetics of ICA in rats following multiple oral administration. Methods ICA and an internal standard coumestrol（ CMT） were extracted from rat plasma using ethyl acetate and separated on a BEH C18（ 50 mm × 2. 1mm,1. 7 μm） column using a gradient mobile phase of acetonitrile containing 0. 1%（ V / V）formic acid and water containing 2 mmol / L ammonium formate at a flow rate of 0. 3m L / min. In negative electrospray ionization mode,multiple reaction monitoring of the precursor-product ion transitions of m / z 675. 6→351. 1 for ICA,267. 0 →211. 1 for CMT was used for the quantification. Plasma was collected after rats were orally administered with ICA at multiple doses of 50 mg / kg. Results The linear calibration curve was achieved in a concentration range of 0. 5-50 ng / m L with a lower limit of quantification of0. 5 ng / m L. The value of intra- and inter-day precision was less than 11. 3% and accuracy fell in the ranges of 94. 3%-98. 7%. The recovery ranged from 81. 3% to85. 2% and the matrix effects from 94. 3% to 103. 2%. After oral administration of ICA to rats,t1 /2was（ 1. 68 ± 0. 29） h,Cmaxwas（ 29. 6 ± 5. 3） ng / m L,tmaxwas（ 1. 00 ± 0. 00） h,AUC0- twas（ 88. 4 ± 13. 9）（ h·ng） / m L. Conclusion The method is specific and accurate,suitable for preclinical pharmacokinetics of ICA.