唾液中皮质醇测定的液相色谱-质谱法  

Determination of Cortisol in Saliva by Liquid Chromatography Tandem Mass Spectrometry

作　　者：刘晓东 乔荷 孟祥娟 王超 丁晓文[1] 牛东升[1] 李珏[1] Liu Xiaodong;Qiao He;Meng Xiangjuan;Wang Chao;Ding Xiaowen;Niu Dongsheng;Li Jue(The Beijing Prevention and Treatment of Hospital of Occupational Disease for Chemical Industry,Beijing 100093,China;Beijing University of Chemical Technology,Beiijng100029,China)

机构地区：[1]北京市化工职业病防治院,100093 [2]北京化工大学,北京100029

出　　处：《中华劳动卫生职业病杂志》2019年第2期143-146,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金："十三五"国家重点研究计划项目(2016YFC0801700);北京市博士后工作经费资助项目(2018-ZZ-037、2018-ZZ-038).

摘　　要：目的建立测定唾液中皮质醇的液相色谱-质谱检测方法。方法取0.5ml唾液样品到2ml离心管,并向其中加入过量氯化钠和1ml乙腈,涡旋震荡3 min,15 000 r/min条件下离心10 min,取上层液体800μl至另一离心管。样品经过真空浓缩仪浓缩并用初始流动相定容到200μl后进仪器分析。利用外标标准曲线法对目标化合物进行定量。结果该方法的线性范围为0.02~5.00 ng/ml,r=0.999 9,方法检出限为0.002ng/ml,方法定量下限为0.02 ng/ml,加标回收率为89.60%~98.60%,批内精密度为1.90%~3.30%,批间精密度为4.20%~9.00%;样品在-20℃条件下可放置14 d;样品中的其他内源性物质不干扰皮质醇的测定。结论该方法前处理简单,灵敏度高,重现性和回收率好,适用于正常人群和职业紧张人群唾液中皮质醇的定量分析。Objective To establish a liquid chromatography tandem mass spectrometry method to determine the cortisol in saliva.Methods Take 0.5 ml saliva sample into a 2ml centrifuge tube,add excess sodium chloride and 1ml acetonitrile to it,then vortex for 3 min,centrifuge for 10 min at 15 000 r/min,and take 800μl of the upper layer to another centrifuge tube.Finally,the sample was concentrated by a vacuum concentrator and brought to 200μl with the initial mobile phase.Then,the sample was analyzed by liquid chromatography tandem mass spectrometry.The target compound was quantified by external standard curve method.Results The linear range of the method was 0.02-5.00 ng/ml,r=0.999 9,the method limit of the detection was 0.002 ng/ml,the method limit of quantitative was 0.02 ng/ml,and the spiked recoveries were 89.60%-98.60%.The intra-assay precision was 1.90%-3.30%,and the inter-assay precision was 4.20%-9.00%;samples could be stored at-20℃for at least 14 days.The determination of cortisol could not be interfered by other endogenous substances in the sample.Conclusion The method is simple in pretreatment,high sensitivity,good reproducibility and good recovery,and it is suitable for the quantitative analysis of cortisol in saliva for normal and occupationally stressed populations.

关 键 词：皮质醇 唾液 液相色谱-质谱 职业应激 

分 类 号：R446.1[医药卫生—诊断学] O657.63[医药卫生—临床医学]