In this study, we report the synthesis and application of a FePt/CNTs nanocomposite as a highly sensitive sensor and novel amide ligand (9,10-dihydro-9,10-ethanoanthracene-11,12-dicarboximido)-4-ethylbenzene-1,2-diol as a mediator for the determination of glutathione (GSH), nicotinamide adenine dinucleotide (NADH) and tryptophan (Trp). The synthesized materials were characterized with different methods such as NMR, IR spectroscopy, TEM, XRD, FESEM, cyclic voltammetry, electrochemical impedance spectroscopy and square wave voltammetry (SWV). The modified electrode exhibited a potent and persistent electron mediating behavior followed by well-separated oxidation peaks of GSH, NADH and Trp. The peak currents were linearly dependent on GSH, NADH and Trp concentrations in the range of 0.08–220, 1.0–400 and 5.0–500 μmol L^–1, with detection limits of 0.05, 0.8 and 1.0 μmol L^–1, respectively. The modified electrode was used for the determination of these compounds in real samples.