液相色谱-串联质谱法及在线样品处理法测定人血浆中依普黄酮的浓度

目的建立在线样品处理技术和液相色谱-串联质谱法测定人血浆中依普黄酮(雌激素抗骨质疏松药物)浓度的方法。方法血浆样品稀释后,直接用HPLC二维系统完成在线预处理和分离。使用前处理柱为Shim-Pack MAYI-ODS柱,流动相为纯水-甲醇梯度配比,分析柱为Dikma Diamonsil C18色谱柱,流动相为0.2%甲酸水溶液-甲醇=15∶85,通过电喷雾离子化三重四级杆质谱,经多反应检测正离子模式进行检测。结果依普黄酮的血药浓度在0.5～250.0 ng.mL-1线性良好(γ=0.999),检测限在0.5 ng.mL-1,提取回收率在81%～88%,日内、日间精密度均在(RSD)≤11.6%,准确度在87.6%～113.7%。结论该法操作快速高效、灵敏准确、重现性好,能够满足临床药代动力学研究的需要。

Determination the concentration of ipriflavone in human plasma by on -line extraction and HPLC- MS/MS

Objective To establish a on-line extraction coupling with HPLC-MS/MS approach to determine ipriflavone in human plasma.Methods Extraction and separation was achieved on the two-dimension HPLC system immediately after plasma samples dilution.The extraction column was Shim-pack MAYI-ODS with the mobile phase consisted mixture of water-methanol(gradient),while the analytical column was Dikma Diamonsil C18with the mobile phase consisted mixture of 0.2%formic acid-methanol(15∶85).Detection was performed on a tandem mass spectrometry with electrospray ionization source in positive ion mode,and multiple reaction monitoring(MRM) mode with the transition.Results The linear range was 0.5-250.0 ng·mL-1 with the correlation coefficient being 0.999,and the limit of quantitation was 0.5 ng·mL-1.The extraction recovery was 81%to 88%,intra-and inter-day precisions(RSD)≤11.6% while accuracies were 87.6% to 113.7%.Conclusion The method demonstrated very rapid,efficient,sensitive,accurate and reproducible,it can satisfy clinical pharmacokinetics study.