Electrooxidation of glutathione (GSH) was studied at the surface of 2,7-bis (ferrocenyl ethyl) fluoren-9-one modified carbon paste electrode (2,7-BFEFMCPE). Cyclic voltammetry (CV), double potential-step chronoamperometry, and differential pulse voltammetry (DPV) were used to investigate the suitability of this ferrocene derivative as a mediator for the electrocatalytic oxidation of GSH in aqueous solutions with various pH. Results showed that pH 7.00 is the most suitable pH for this purpose. At the optimum pH, the oxidation of GSH at the surface of this modified electrode occurs at a potential of about 0.410 V versus Ag|AgCl|KClsat. The kinetic parameters such as electron transfer coefficient, alpha = 0.61, and rate constant for the chemical reaction between GSH and redox site in 2,7-BFEFMCPE, k (h) = 1.73 x 10(3) cm(3) mol(-1) s(-1), were also determined using electrochemical approaches. Also, the apparent diffusion coefficient, D (app), for GSH was found to be 5.0 x 10(-5) cm(2) s(-1) in aqueous buffered solution. The electrocatalytic oxidation peak current of GSH showed a linear dependence on the glutathione concentration, and linear calibration curves were obtained in the ranges of 5.2 x 10(-5) M to 4.1 x 10(-3) M and 9.2 x 10(-7) M to 1.1 x 10(-5) M with cyclic voltammetry and differential pulse voltammetry methods, respectively. The detection limits (3 sigma) were determined as 1.4 x 10(-5) M and 5.1 x 10(-7) M for the CV and DPV methods, respectively. This method was also examined as a selective, simple, and precise new method for voltammetric determination of GSH in real sample such as hemolysed erythrocyte.