Determination of testosterone and cortisol in saliva of healthy human by liquid chromatography-tandem mass spectrometry / 第二军医大学学报

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.