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目的:建立以液-质联用法测定人血浆中脱氢表雄酮硫酸酯(DHEAS)浓度的方法。方法:选择雌酮硫酸酯(ES)作为内标。血浆使用乙腈沉淀除去蛋白,经固相萃取、水解及衍生化后,以高效液相色谱-质谱检测器测定DHEAS的浓度。色谱柱为AgilentSBC18,柱温为40℃,流动相为乙腈-水(梯度洗脱);采用大气压化学离子源,负离子检测方式,用于选择离子检测(SIM)定量分析的离子为m/z490.0(DHEAS)和m/z472.1(ES)[M-H]-。结果:DHEAS血药浓度在250.0～320.0ng.mL-1范围内线性关系良好(r=0.9994);模拟白蛋白样品的提取回收率为71.1%～78.9%,相对回收率为98.3%～101.4%,日内、日间RSD均<10%。测得15名正常老年志愿者的平均血浆DHEAS浓度为(981.6±353.4)ng.mL-1。结论:本方法简便、实用、准确、灵敏,适合测定人血浆中DHEAS的浓度。
Objective: To establish a method for the determination of DHEAS in human plasma by liquid chromatography - mass spectrometry. Methods: Estrone sulphate (ES) was selected as internal standard. The plasma was treated with acetonitrile to remove protein. After solid-phase extraction, hydrolysis and derivatization, the concentration of DHEAS was determined by high performance liquid chromatography-mass spectrometry. The column was Agilent SBC18. The column temperature was 40 ° C and the mobile phase was acetonitrile-water (gradient elution). The atmospheric pressure chemical ion source and negative ion detection mode were used for the ion selective ion detection (SIM) for m / z 490. 0 (DHEAS) and m / z 472.1 (ES) [MH] -. Results: The linear range of DHEAS was in the range of 250.0-320.0 ng.mL-1 (r = 0.9994). The recovery of simulated albumin was 71.1% -78.9% and the relative recovery was 98.3% -101.4% , Day, day RSD <10%. The average plasma DHEAS concentration of 15 normal elderly volunteers was (981.6 ± 353.4) ng.mL-1. Conclusion: The method is simple, practical, accurate and sensitive and suitable for determining the concentration of DHEAS in human plasma.