LC-MS法检测人血浆中双氯芬酸的浓度

Determination of diclofenac in human plasma by LC-MS

目的：建立LC-MS测定人血浆中双氯芬酸浓度的方法。方法：待测血浆样品经乙酸乙酯萃取后,用HPLC-MS法进行检测,以格列吡嗪为内标。色谱柱为Agilent ZORBAX SB-C18（150 mm×4.6 mm,5μm）,流动相为10 mmol.L-1醋酸铵溶液-甲醇（35∶65）,流速为0.5 mL.min-1;质谱采用电喷雾离子化（ESI）方式和选择性离子检测（SIM）模式,检测离子为负离子。结果：本方法线性范围为3.936 0~492.000 0 ng.mL-1,最低定量浓度3.936 0 ng.mL-1,准确度、精密度以及稳定性均符合有关要求。结论：该方法简便、准确,灵敏度高,专属性强,适用于人血浆中双氯芬酸浓度的测定。

Objective：To establish a liquid chromatography-mass spectrometry（LC-MS） method for determination of diclofenac in human plasma.Methods：The drug and an internal standard（glipizide） were extracted from plasma with ethyl acetate.The compounds were separated on a Agilent ZORBAX SB-C18（150 mm×4.6 mm,5 μm） column by Agilent LC-MS with 0.01 mol·L-1 ammonium acetate-methanol（35∶65） as the mobile phase at a flow rate of 0.5 mL·min-1.The detection was carried out by means of electrospray ionization mass spectrometry in positive ion mode.Selective ion（SIM） mode was used.Results：The assay had the calibration range from 3.936 0 to 492.000 0 ng·mL-1 and the lower limit of quantification（LOQ） was 3.936 0 ng·mL-1 for diclofenac.The accuracy,precision（intra-day and inter-day）,sensitivity,specificity and stability（ambient temperature,freeze/thaw,freeze storage） of the method fulfilled the analytical validation criteria of the guideline.Conclusion：The method is simple,accurate,sensitive and specific for determination of diclofenac in human plasma.