In the current research, a method of electrophoretic determination of steroid hormones in urine sample with the use of dispersive liquid-liquid microextraction in imidazolium ionic liquid (IL) C₈MImBF₄ during the sample preparation step was proposed. Ionic liquids С₆MImNTf₂, С₆MImBF₄ С₈MImBF₄ were examined as extractants while acetone, methanol and acetonitrile were tested as dispersants. It was found that the IL nature influences the extraction of steroid hormones, i.e. the extraction degrees were raised when IL with BF₄ ^- anion was used instead of NTf₂, and when the length of alkyl radical in imidazolium cation was increased. It was also established that pH of water phase did not affect the steroid hormones extraction. The highest extraction degree values of analytes (69-93%) were obtained under the following condition: addition of 150 mkl of IL C₈MImBF₄ dissolved in 0.5 ml of acetone to 5 ml aqueous sample. The developed sample preparation technique was successfully applied to the determination of corticosteroid in urine. It was found that the high concentration of salt in the sample matrix favors the mass transfer of analyte to organic phase, and the extraction degrees in IL were increased. The preconcentration factors in sample preparation were 23-30; the achieved limits of quantification were 8-12 ng/ml.