A high-performance liquid chromatographic method for the determination of diazolidinyl urea (DU) in a cream formulation is described. The aqueous phase of the emulsion was separated by centrifugation, removed, filtered, diluted and applied onto the HPLC system. DU was detected by ultraviolet absorption at a wavelength of 214 nm. The calibration curve was linear over the range of 79-553 micrograms/ml, and identical when determined on consecutive days. The relative standard deviation for repeat determinations was less than 0.5%. Recoveries were 97.74-101.72%. This analytical method is useful for quantitation of DU in cream formulations.