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目的建立超高效液相色谱-串联质谱法(UHPLC-MS/MS)同时测定大鼠血浆中淫羊藿素(ICT)及其葡萄糖醛酸代谢物(GICT)浓度,并用于定量研究ICT在大鼠体内的代谢。方法采用乙腈提取血浆中ICT和GICT,以香豆雌酚(CMT)为内标,在C18色谱柱上以乙腈-水-甲酸铵-甲酸梯度洗脱分离;在电喷雾离子化电离源上以选择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m/z 367.1→297.1(ICT),m/z 543.3→367.1(GICT)和m/z 267.0→211.1(CMT)。大鼠腹腔注射10 mg/kg ICT,采集不同时刻血样并测定ICT和GICT浓度。结果 ICT和GICT的线性范围分别为0.2~20 ng/ml和2~200 ng/ml,定量下限分别为0.2 ng/ml和2ng/ml。ICT和GICT的精密度(RSD)在10.3%以内,准确度在95.1%~103.7%,提取回收率为89.1%~92.4%,基质效应为93.2%~104.2%。结论 UHPLC-MS/MS法专属性好、准确度高,适用于ICT在大鼠体内代谢研究,2 h时GICT达到最大浓度。ICT在大鼠体内迅速代谢成葡萄糖醛酸Ⅱ相代谢物GICT。
Objective To establish simultaneous determination of icariin (ICT) and glucuronic acid metabolites (GICT) in rat plasma by ultra performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) Rat metabolism. Methods The ICT and GICT in plasma were extracted by acetonitrile. The coumarin (CMT) was used as the internal standard and eluted on a C18 column with gradient elution of acetonitrile-water-ammonium formate-formic acid. On the electrospray ionization ionization source Negative ion detection was selected for reaction monitoring. The ion reactions for quantitative analysis were m / z 367.1 → 297.1 (ICT), m / z 543.3 → 367.1 (GICT) and m / z 267.0 → 211.1 (CMT), respectively. Rats were injected intraperitoneally with 10 mg / kg ICT, blood samples were taken at different time points and the concentrations of ICT and GICT were measured. Results The linear ranges of ICT and GICT were 0.2 ~ 20 ng / ml and 2 ~ 200 ng / ml, with the lower limit of quantitation of 0.2 ng / ml and 2 ng / ml, respectively. The precision of ICT and GICT was less than 10.3%, with the accuracy of 95.1% ~ 103.7%. The recovery rates of ICT and GICT were 89.1% ~ 92.4% and the matrix effects were 93.2% ~ 104.2%. Conclusion The UHPLC-MS / MS method has good specificity and high accuracy, and is suitable for the metabolic studies of ICT in rats. The maximum concentration of GICT reached at 2 h. ICT is rapidly metabolized in rats to glucuronic acid phase II metabolite GICT.