HPLC-MS/MS测定血浆中的牡荆素及其大鼠体内药动学研究

Determination of Vitexin in Plasma by HPLC-MS/MS Method and Its Pharmacokinetics in Rats

目的:建立高效液相色谱-质谱/质谱联用法(HPLC-MS/MS)测定血浆中牡荆素的方法,并用于大鼠药动学研究。方法:采用Capcell Pak C18柱(50 mm×2.0 mm,I.D.,5μm),柱温25℃,流动相为甲醇∶水(95∶5,0.1%甲酸),流速为0.2 mL/min。用负离子电喷雾电离源,多反应方式检测,选择监测的离子反应为m/z 431～311(牡荆素)和m/z 269～225(大黄素,内标)。结果:牡荆素在0.5～2 000 ng/mL线性关系良好,相关系数为0.9960,最低定量限为0.5 ng/mL。加样回收率76.1%～89.0%,日内日间精密度RSD均小于11%。结论:该方法分析速度快、操作简单、灵敏度高,可用于大鼠血浆中牡荆素的浓度测定及药动学研究。

Objective：To establish a HPLC-MS/MS method for the determination of vitexin in rat plasma and its pharmacokinetics.Methods：The HPLC-MS/MS method used Capcell Pak C18 column（50 mm×2.0 mm I.D.,5 μm）.The mobile phase was methanol and water（95∶ 5,V/V,containing 0.1% formic acid） at a flow rate of 0.2 mL/min.Electrospray ionization（ESI） in negative ion mode and multiple reaction monitoring（MRM） was used for the quantification of vitexin with a monitored transitions m/z 431→311 for vitexin and m/z 269→225 for internal standard（I.S.,emodin）.Results：Linear calibration curves were obtained over the concentration range of 0.5~2000 ng/mL（r=0.9960） with the lowest limit of quantification（LLOQ） of 0.5 ng/mL.The recovery was in the range of 76.1%~ 89.0%.The relative standard deviations for the intra-day and inter-day validation were less than 11%.Conclusion：The method is simple,accurate,fast,sensitive and suitable for the pharmacokinetic study of vitexin in rats.