Differential pulse cathodic adsorptive stripping voltammetric method was developed for the determination of prednisolone based on electrochemical reduction of the drug at a hanging mercury drop electrode (HMDE) in 0.04 M Britton-Robinson buffer pH 3.5. The peak current varied linearly over the concentration range 7.21-144.2 ng/ml prednisolone with limit of detection and limit of quantification of 3.95 and 13.17 ng/ml prednisolone, respectively. The proposed method was successfully applied for the determination of the drug in commercial tablets and spiked human urine samples.