The preparation and electrochemical performance of the carbon nanotube-paste electrode modified with salophen complexes of cobalt(III) perchlorate, with various substituents on the salophen ligand, as well as their electrocatalytic activity toward the oxidation of N-acetylcysteine (NAC) is investigated. Several Schiff base complexes containing various nucleophilic and electrophilic functional groups were prepared, and their electrochemical characteristics for the electro-oxidation of NAC were evaluated using cyclic and differential pulse voltammetry (CV and DPV). The results revealed, the modified electrodes show an efficient and selective electrocatalytic activity toward the anodic oxidation of NAC among biologically important compounds in buffered solutions at pH of 7.0. The best voltammetric responses were obtained for a carbon-paste electrode (CPE) modified with a salophen complex containing para-methoxy groups on its salicylaldehyde ring. The analytical response of the modified electrode for response to other sulfhydryl compounds (e.g., cysteine, penicillamine, captopril and mercaptopropionyl glycine) in comparison to NAC was investigated by CV and DPV methods. The DPV method was applied as a sensitive method for the quantitative detection of the trace amounts of NAC. A linear dynamic range from 1 × 10−7 to 1 × 10−4 M with calibration sensitivity of 0.0646 μA/μM is resulted for NAC in DPV measurements. The detection limit was 5 × 10−8 M, which is remarkably lower than those reported previously for NAC using other modified electrodes. The results of voltammetric determinations show a very good reproducibility, and the R.S.D. for the slope of the calibration curve, based on 10 measurements in a period of two months, was <3.9%. The detection system provides very stable electrochemical responses toward NAC, makes it very suitable for using in pharmaceutical and clinical measurements.