LC-MS法测定人血浆中脱氢表雄酮硫酸酯的浓度

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。

Determination of Dehydroisoandrosterone Sulfate in Human Plasma by LC- MS

OBJECTIVE： To determine the concentration of dehydroisoandrosterone sulfate （DHEAS） in human plasma by LC-MS. METHODS： With estrogen sulfate （ES） served as an internal standard,the plasma samples were deproteinized with acetonitrile, extracted by solid phase, hydrolyzed and derivatized. Then the concentration of DHEAS was determined by HPLC- MSD on Agilent SB C18 with column temperature kept at 40℃ . The mobile phase consisted of acetonitrile - water （in gradient elution）. Atmosphere pressure chemical ion source in negative ion detection model was employed. The ions selected for SIM （selected ion monitoring） quantitative analysis included m/z 490.0 （DHEAS） and m/z 472. I（ES）M-H]^-. RESULTS： The linear range of DHEAS was 250.0--320.0 ng· mL^-1（ r = 0.999 4） . The extraction recovery of the simulated human albumin samples ranged from 71.1%- 78.9% and its relative recovery ranged from 98.3%- 101.4%. Both the intra- day RSD and inter-day RSD were less than 10%. The mean concentration of DHEAS in 15 health aged male volunteers was （981.6±353.4） ng · mL^-1. CONCLUSION： The method is simple, practical, accurate and sensitive, and it is applicable for the determination of plasma concentration of DHEAS.