超高效液相色谱-串联质谱法测定大鼠血浆中淫羊藿素

UPLC-MS/MS method for the determination of icaritin in rat plasma

目的建立超高效液相色谱-串联质谱法测定大鼠血浆中淫羊藿素（ICT）浓度，并用于研究大鼠静脉注射ICT的药动学。方法采用乙腈提取血浆中ICT，以香豆雌酚（CMT）为内标，在BEHc18色谱柱（2．1mm×50mm，1．7μm）上以流动相乙腈-水-甲酸铵-甲酸梯度洗脱，流速0．3ml／min，柱温40℃，分析时间6．5min；在电喷雾离子化电离源上以多反应监测方式进行负离子检测，用于定量分析的离子反应分别为mgz367．1→297．1（ICT）和m／z267．0→211．1（CMT）。大鼠静脉注射10mg／kgICT，采集不同时刻血样并测定药物浓度。结果ICT的线性范围为0．5～20ng／ml，方法定量下限为0．5ng／ml。ICT日内和日间精密度（RSD）在8．1％以内，准确度在95．3％～99．3％，提取回收率为90．1％-92．1％，基质效应为97．5％～101．2％。大鼠静脉注射ICT的t1／2为（1．13±0．32）h，表观分布容积为（4．54±0．27）L／kg，清除率为（2．91±0．68）L／（h·kg）。结论该方法速度快、专属性好、准确度高，适用于ICT临床前药动学研究。

Objective To establish an ultra-performance liquid chromatography- tandem mass spectrometry （UPLC-MS/MS） method for the quantification of icairitin （ICT） , and investigate pharmacokineties of ICT in rats following a single intravenous administration. Methods ICT and an internal standard coumestrol （CMT） were extracted from rat plasma using acetonitrile and separated on a BEH C18 column （2. 1 mm × 50 mm, 1.7 p,m） using a gradient mobile phase of acetonitrile, water, ammonium formate and formic acid at a flow rate of 0.3 ml/min. At negative electrospray ionization mode, multiple reaction monitoring of the precursor-product ion transitions of m/z 367.1→297.1 for ICT, 267.0→211.1 for CMT was used for the quantification. Plasma was collected after rats were intravenously administered with ICT at a single dose of 10 mg/kg. Results The linear calibration curve was obtained in a concentration range of 0.5 - 20 ng/ml with a lower limit of quantification of 0.5 ng/ml. The value of intra- and inter-day precision was less than 8.1% and accuracy fell in the ranges of 95.3% -99. 3%. The recovery ranged from 90. 1% to 92. 1% and the matrix effects from 97.5% to 101.2%.After intravenous administration of ICT to rats, t1/2was（ 1. 13 ± 0.32） h, V was（4.54 ± 0.27） L/kg,and CL was（2.91 ± 0.68） L/（h.kg）. Conclusion The method was rapid, specific and accurate, suitable for preclinical pharmacokinetics of ICT.