HPLC-MS/MS法测定人血浆中普罗布考浓度

目的建立高效液相色谱-串联质谱(HPLC-MS/MS)技术测定人血浆中普罗布考浓度的方法。方法血浆经乙酸乙酯∶二氯甲烷(1∶1,V/V)提取后,采用HPLC-MS/MS测定。以ultimate CN(50mm×4.6mm,5μm)为色谱柱,乙腈∶水∶氨水=97∶3∶0.05(甲酸调pH=7.20)为流动相进行分离。质谱采用负离子模式检测,以大黄素甲醚为内标,检测离子515.5→236.1(普罗布考)和283.0→239.9(内标)。结果普罗布考在2.5～6000ng/mL范围内线性关系良好,最低定量限为2.5ng/mL,方法回收率在93.02%～104.12%之间,日内、日间精密度分别小于4.67%及5.72%。结论本研究所建立的方法准确、灵敏,可用于普罗布考血药浓度测定及药代动力学研究。

Determination of Probucol in Human Plasma by HPLC-MS/MS

Objective To develop a specific and sensitive method for the determination of probucol in human plasma using HPLC-MS/MS technique.Methods Probucol were extracted from plasma by ethyl ether∶dichloromethane(1∶1,V/V),with physcion as an internal standard.The analytes went through the column of ultimate CN(50 mm×4.6 mm,5 μm) with mobile phase acetonitrile∶water∶ammonia water=97∶3∶0.05(adjusted pH=7.2 with formic acid).Probucol was analyzed with a negative mode.The ion pairs being detected were 515.5→236.1(probucol) and 283.0→239.9 physcion,respectively.Results The established method was able to determine probucol in human plasma over the range of 2.5-6000 ng/mL,with a method recovery ranging from 93.02% to 104.12%.The intra and inter day variances were below 4.67% and 5.72%.Conclusion The HPLC-MS/MS method for analyzing probucol was validated.It is sensitive and suitable for pharmacokinetic studies of probucol.