UPLC-MS/MS法测定人血浆中的醋酸甲地孕酮

Determination of Megestrol Acetate in Human Plasma by UPLC-MS/MS

目的:建立超高效液相色谱-串联质谱法(UPLC-MS/MS)测定人血浆中醋酸甲地孕酮的浓度。方法:采用蛋白沉淀法预处理血浆样本,沉淀剂为甲醇。采用Waters BEH C_(18)色谱柱(50 mm×2.1 mm,1.7μm),流动相A为含0.1%甲酸和10 mmol·L^(-1)乙酸铵的水溶液,流动相B为甲醇,梯度洗脱,流速0.3 ml·min^(-1),柱温40℃,进样量1μl。结果:醋酸甲地孕酮的线性范围为1~400 ng·ml^(-1);定量下限为1 ng·ml^(-1);批内精密度RSD均≤6.50%,批间精密度RSD均≤5.86%;醋酸甲地孕酮的提取回收率≥90.93%。结论:所用方法简便快速、准确度高、灵敏度好,适用于测定人血浆中甲地孕酮的浓度。

Objective:To establish a UPLC-MS/MS method for determining megestrol acetate in human plasma.Methods:The samples were treated by protein precipitation with methanol.The chromatographic column was Waters BEH(50.0 mm×2.1 mm,1.7μm).Megestrol acetate was determined by gradient elution with 0.1%aqueous formic acid containing 10 mmol·L^(-1)ammonium acetate as mobile phase A and methanol as mobile phase B.The flow rate was 0.3 ml·min^(-1)with the column temperature at 40℃,and the injection volume of 1μl.Results:The standard curve was within the range of 1-400 ng·ml^(-1).The LLOQ was 1 ng·ml^(-1).All the RSDs of intra-batch precision were not more than 6.50%,and all the RSDs of inter-batch precision were not more than 5.86%.The recovery of megestrol acetate was not less than 90.93%.Conclusion:The method is easy,rapid,accurate and sensitive.It is suitable for determining megestrol acetate in human plasma.