HPLC-MS/MS法测定琥乙红霉素颗粒体内活性代谢物

HPLC-MS/MS determination active metabolite in vivoof erythromycin ethylsuccinate granules

目的:建立琥乙红霉素人体内活性代谢物红霉素的HPLC-MS/MS分析方法。方法:选用克拉霉素为内标,血浆样品经正己烷-二氯甲烷-异丙醇(300∶150∶15)提取处理,通过检测活性代谢物红霉素的浓度来监测琥乙红霉素体内行为。色谱条件Waters C18色谱柱(4.6 mm×250 mm,5μm),甲醇-水(80∶20,含0.5%甲酸)为流动相,流速0.6 mL·min-1,采用HPLC-MS/MS检测系统(SRM模式),质谱采用电喷雾电离源(ESI),红霉素选择检测离子对为m/z 734→m/z 158,内标克拉霉素选择检测离子对为m/z 748→m/z 158。结果:血浆中红霉素检测方法的线性范围为9.724～2431.0 ng·mL-1,最低检测限可达9.724 ng.mL-1。血浆中红霉素的平均回收率为77.6%～80.0%;日内、日间RSD均小于11%。结论:本法灵敏、准确、选择性高,可用于监测琥乙红霉素的体内行为。

Objective： To establish an HPLC-MS-MS method for the determination of erythromycin as an active metabolite of erythromycin ethylsuccinate in human plasma.Methods： The plasma was extracted by n-hexane-dichloromethane-isopropyl alcohol（300∶150∶15） and the internal standard was clarithromycin.Erythromycin was detected to monitor the erythromycin ethylsuccinate in plasma.The Waters C18 column（4.6 mm×250 mm,5 μm） was used,the mobile phase was a mixture of methanol-waters（80∶20,containing 0.5% formic acid）,the flow rate was 0.6 mL·min-1.HPLC-MS/MS system（SRM mode） and ESI was used.The ion of monitor：m/z 734→m/z 158（erythromycin）,m/z 748→m/z 158（clarithromycin）.Results： The linear range of the standard curve of erythromycin was 9.724-2431.0 ng·mL-1 and the limit of detection was 9.724 ng·mL-1.The recovery of erythromycin was 77.6%-80.0%,intra-day and inter-day RSDs were less than 11%.Conclusion：The method had high sensitivity,good selectivity and precision.It is suitable for pharmacokinetic studies of erythromycin ethylsuccinate.