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液相色谱-串联质谱法测定健康人唾液中睾酮和皮质醇的含量
顾伟锋1,梁凤英2,刘林菁2,高守红2*
0
(1. 海军上海舰艇岸勤部门诊部, 上海 200083;
2. 海军军医大学(第二军医大学)长征医院药学部, 上海 200003
*通信作者)
摘要:
目的 建立一种方便、可靠和灵敏的液相色谱-串联质谱(LC-MS/MS)方法用于检测健康人唾液中2种内源性激素(睾酮和皮质醇)的水平。方法 在Agilent 6410A串联质谱仪上采用正离子检测模式,以卡马西平为内标,唾液样品经OMGEA NANOSEP 10K超滤管21 912.8×g超滤10 min后,在Agilent ZORBAX SB-C18(3.5 μm,2.1 mm×100 mm)色谱柱上采用等度洗脱进行色谱分离。流动相为乙腈:0.1%甲酸水溶液(60:40);流速为0.3 mL/min;进样量为10 μL;柱温为25℃;每个样品的分析周期为3 min。结果 睾酮和皮质醇线性关系良好,其相关系数r>0.990,日内与日间精密度均<15%。结论 该方法可以用于测定唾液中睾酮和皮质醇的含量,为迅速简便地检测体内激素水平提供参考。
关键词:  液相色谱-串联质谱法  唾液  睾酮  皮质醇  内源性激素
DOI:10.16781/j.0258-879x.2019.01.0104
投稿时间:2018-05-29修订日期:2018-11-15
基金项目:上海市临床药学重点专科建设项目(2016-40044-002),上海市卫生计生系统重要薄弱学科建设项目(2016ZB0303).
Determination of testosterone and cortisol in saliva of healthy human by liquid chromatography-tandem mass spectrometry
GU Wei-feng1,LIANG Feng-ying2,LIU Lin-jing2,GAO Shou-hong2*
(1. Department of Outpatient, Shanghai Naval Vessel Service Department, Shanghai 200083, China;
2. Department of Pharmacy, Changzheng Hospital, Naval Medical University(Second Military Medical University), Shanghai 200003, China
*Corresponding author)
Abstract:
Objective To establish a convenient,reliable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of two kinds of endogenous hormones (testosterone,cortisol) in the saliva of healthy human.Methods A positive ion mode was performed on the Agilent 6410A mass spectrometer. Carbamazepine was used as the internal standard.The saliva samples were centrifuged at 21 912.8×g for 10 min with OMGEA NANOSEP 10K ultrafiltration tubes,and chromatographic separation was achieved on an Agilent ZORBAX SB-C18 column with isocratic elution.The mobile phase was acetonitrile:0.1% formic acid awter solution (60:40);flow rate was 0.3 mL/min;injection volume was 10 μL;column temperature was 25℃;and the analysis period of each sample was 3 min.Results Testosterone and cortisol had good linear relationship with correlation coefficient (r) being both >0.990.Inter-and intra-day precision was both less than 15%.Conclusion This method can be used to determine the contents of testosterone and cortisol in saliva,which provides a reference for rapid and simple detection of hormone levels in vivo.
Key words:  liquid chromatography-tandem mass spectrometry  saliva  testosterone  cortisol  endogenous hormones