Abstract: Objective?To develop a method for simultaneous determination of adrenaline, noradrenaline, cortisone and cortisol in plasma using HPLC/MS/MS. Methods?Sample proteins were precipitated with acetonitrile and the sample solution was injected into HPLC/MS/MS after centrifugation at 15 000 r/min for 5 min. Electrospray ionization (ESI) and the positive ion detection were applied with a multiple reaction monitoring (MRM) mode for quantitative analyses. Results?Under the optimal conditions, good linearity (r>0.999) was observed in the range of 0.02-200.00 ng/mL of target compounds. The detection limit reached 4.13 pg/mL, 4.64 pg/mL, 4.29 pg/mL and 4.52 pg/mL for adrenaline, noradrenaline, cortisone and cortisol respectively. The inter-day and intra-day precisions ranged from 1.19%-5.42% and 2.16%-6.04% respectively. Satisfied results were achieved using human plasma samples, with a spiked recovery in the range of 80.0%-109.0% and a relative standard deviation of 3.93%-7.57%. Conclusion?The proposed method is quick, sensitive and suitable for batch analyses of plasma samples.