Study on steroids detection based on dual derivatization couple with tandem mass spectrometry
Objective:To establish a dual derivatization method combined with tandem mass spectrometry for the simultaneous determination of 18 different steroid hormones in human serum.Methods:Serum samples were trea-ted with hydroxylamine and 1,2-dimethylimidazole-5-sulfonyl chloride for derivatization,and the resulting compounds were analyzed by liquid chromatography-tandem mass spectrometry in positive ion selected reaction monitoring(SRM)mode.The Kinetex ® C8 column(100 mm ×2.1 mm,2.6 μm)was used for the separation.Mobile phase A(0.1％acetic acid in water)and mobile phase B(0.1％acetic acid in methanol)with gradient elution(0-0.5 min,35％B;0.5-5 min,35％B→100％B;5.0-5.1 min,100％B→35％B;5.1-7 min,35％B)at the flow rate of 0.4 mL·min-1 were applied.Column temperature was set at 40 ℃.Injection volume was 20 μL and collection time was 6 min.Results:The linearity correlation coefficients for all 18 steroid hor-mones were greater than 0.99,the recovery rates ranged from 85％to 115％,and the precision RSD was less than 15％.This method was successfully applied to the analysis of serum samples from 156 healthy subjects(75 males and 61 females),and reference intervals were established.Conclusion:This method can be used to simul-taneously determine 18 types of steroid hormones,such as pregnenolone,17-hydroxypregnenolone,progester-one,17-hydroxyprogesterone,corticosterone,cortisol,11-deoxycorticosterone,21-deoxycorticosterone,aldo-sterone,testosterone,androstanedione,dehydroepiandrosterone,sulfated dehydroepiandrosterone,adrenocortico-tropin,18-hydroxicorticotropin,estrone,estradiol,and estriol using non-solid-phase extraction.
steroid hormoneliquid chromatography-tandem mass spectrometryhydroxylaminedual derivatiza-tionendocrine disordershigh throughput
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