高效液相色谱-质谱联用法测定人血浆中依普利酮浓度

Determination of eplerenone in human plasma by HPLC-MS/MS

目的建立高效液相色谱-质谱联用法测定人血浆中依普利酮的浓度的方法,并用于依普利酮片在中国成年健康受试者的药代动力学研究。方法用蛋白沉淀法处理血浆,色谱柱:C_(18)色谱柱(4.6 mm×50 mm,5μm),流动相:甲醇-乙腈-1 mmol·L^(-1)乙酸铵水溶液,梯度洗脱,流速:1.0 m L·min^(-1),柱温:40℃,在多反应监测模式(MRM)下,用电喷雾离子化源,正离子扫描模式检测。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。结果依普利酮血浆浓度在5.00~2000.00 ng·m L^(-1)线性关系良好,标准曲线的回归方程为y=2.95×10^(-2)x+1.46×10^(-2)(r=0.997 2),定量下限为5.00 ng·m L^(-1),低、中、高3个质量浓度的质控样品日内、日间RSD均<10%,3种质量浓度质控样品的提取回收率在99.76%~100.87%,稳定性良好。结论该法准确、灵敏度高、重现性好,可用于人体中依普利酮血浆浓度的测定。

Objective To establish a high performance liquid chromatography-tandem mass spectrometry method(HPLC-MS/MS） for the quantification and pharmacokinetic of eplerenone in human plasma.Methods The plasma was treated with protein precipitation,chromatographic column： Waters Symmetry C（18） column(4. 6 mm × 50 mm,5μm）,mobile phase： methanol-acetonitrile-1 mmol·L-1 ammonium acetate,gradient elution,column temperature： 40 ℃,electrospray ionization(ESI） source was applied and operated in multiple reaction monitoring(MRM） mode via positive ionization. The specificity,lower limit of quantitation and standard curve, precision and recovery rate, matrix effect as well as the stability were investigated. Results A good linearity of eplerenone was obtained in the concentration range of 5. 00-2000. 00 ng·m L-1,with the lower limit of quantitation of 5. 00 ng ·m L-1. The method validation met the criteria for bio-samples determination. The standard curve of bicalutamide was y = 2. 95 × 10-2 x ＋ 1. 46 × 10-2(r =0. 997 2）. Intra-day and inter-day RSD of low,medium and high quality control samples were all less than 10%,the average recoveries of them were between 99. 76%--100. 87%, and stability was good.Conclusion The method was simple,rapid,sensitive and accurate,specific,suitable for the determination of eplerenone in human plasma.