液相色谱串联质谱法测定人血清中硝苯地平的浓度

Determination of nifedipine in human serum by HPLC-MS/MS

目的建立液相色谱串联质谱法测定人血清中硝苯地平浓度。方法血清样品用甲醇沉淀蛋白,内标为尼群地平,色谱柱为Kromasil C18(4.6mm×150mm,5μm),柱温为30℃,流动相为甲醇-水(90:10,V/V),流速为0.5ml·min-1。质谱采用ESI离子源负离子检测,定量分析的离子对为:m/z345.1/122.0(硝苯地平),m/z359.2/122.0(内标尼群地平)。结果硝苯地平在0.5μg·L-1～50μg·L-1范围内线性关系良好(r=0.9987),批内批间RSD均小于15%,提取回收率为64.06%～77.10%。结论该方法快速,灵敏,准确,可用于硝苯地平的临床药动学研究。

Aim To develop and validate an HPLCMS/MS for the determination of nifedipine in human serum.Methods Nitrendipine was added as an internal standard.The sample preparation included a simple deproteinization step with methanol.Chromatographic separation was achieved on a Kromasil C18 column（4.6mm×150mm,5μm） at 30℃ with the mobile phase consisted of methanol-H2O（90：10,V/V）.The mobile phase was eluted at a flow rate of 0.5 mL ·min-1.Tandem mass spectrometer equipped with electrospray ionization source was applied and operated in the negative ion mode.Multiple reaction monitoring using the precursor to produce on combinations of m/z 345.1/122.0 and m/z 359.2/122.0 was performed to quantify nifedipine and nitrendipine,respectively.Results The calibration curves were linear over the range of 0.5μg·L-1～50μg·L-1（r=0.9987）.RSD of intra-batch and inter-batch were less than 15%,extraction recovery of three level concentrations were between 64.06%～77.10%.Conclusion The method is shown to be convenient,accurate,sensitive,and suitable for clinical pharmacokinetic study of nifedipine.