Electrochemical determination of metallothionein (MT) is widely used for environmental studies. This article describes the development and optimization of the procedure for the quantification of metallothionein by square wave cathodic stripping voltammetry. The determination is based on the complexation of cisplatin and MT and the subsequent reduction of the complexes at the electrode. In order to achieve the highest possible sensitivity and resolution of the peak, an optimization of the experimental parameters has been carried out using experimental design methodology (response surface). Seven chemical and physical parameters, namely, pH, cisplatin concentration, buffer concentration, deposition potential, square wave frequency, amplitude of pulse, and step potential, have been optimized to give 9.0, 5.9 microM, 0.65 M, -0.2 mV, 229 Hz, 46 mV, and 2 mV, respectively. Method characterization has been performed, leading to a detection limit of 0.1 microg L(-1). Quantification of MT in polychaetes and comparison with the modified Brdicka procedure were also carried out.