The results indicated that this method showed a wide linearity range (0.1 - 100 ng / mL for cortisol and 0.025 - 100 ng / mL for testosterone), good linearity (r2 > 0.999), low limit of quantification (LLOQ values of cortisol and testosterone were 0.1 ng / mL and 0.025 ng / mL, respectively), good recovery (90.4% - 108.0%), suitable repeatability (CV values were in the range of 2.7% - 14.8%), good stability in different conditions.