The aim of this research was to standardize an high performance liquid chromatographic method for the determination of steroid hormones contained in commercially available pharmaceutical preparations. A Merck LiChrospher®100 RP–18 (5 μm) in LiChroCART®(125-4) column, a rotative valve injector (20 μL loop), ambient temperature, a mobile phase consisting of water-methanol and UV detection at 254nm and 212nm make possible the quantitative determination of dexamethasone acetate, prednisone, ethynylestradiol and norgestrel contained in pharmaceutical preparations.