A kinetic fluorometric method was proposed for the quantitative determination of epinephrine (EP) in human plasma samples with the aid of second-order calibration methods based on alternating trilinear decomposition (ATLD) and alternating fitting residue (AFR) algorithms. It was based on that EP could be gradually converted to a highly fluorescent intermediate product by an oxidation reaction, and further to a non-fluorescent degradation product (o-quinone). These methodologies fully exploit the second-order advantage of the employed three-way kinetic fluorescence data, allowing the concentrations of EP to be quantified even in the presence of uncalibrated interferences. The average recoveries obtained from ATLD and AFR with a factor number of 2 (N = 2) were 100.7 +/- 3.3 and 100.4 +/- 2.2%, respectively. In addition, elliptical joint confidence region (EJCR) tests as well as figures of merit (FOM) were employed to evaluate the accuracy of the two algorithms.