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.