A simple and sensitive method for the determination of steroids using micellar liquid chromatography is described. The steroids, including hydroxycorticosterone. corticosterone, northisterone, testosterone, mexdroprogesterone acetate and progesterone, were separated by reversed-phase using a micelles mobile phase following UV detection at 245 nm. The parameters affecting retention of the test solutes such as the concentration of sodium dodecyl sulfate (SDS) and n-butanol-1 in the mobile phase were investigated. It was found that the retention of the solutes was dependent on the composition of mobile phase. The linear calibration plots range from 0.1 to 10 μg ml−1in mobile phase containing 5.0 × 10−2mol l−1SDS/9 % n-butanol-1 at pH 6.0, and the detection limit in order of 0.1 μg ml−1was obtained. The proposed method was used for the determination of steroids in urine using direct injection of samples without previous treatment.