Abstract
A 23 factorial design has been used in an isocratic HPLC system to study the effect of methanol and acetonitrile composition of the eluents as well as different columns on the retention and resolution of steroids. By factorial design and computer simulation, six steroids of sex hormones and contraceptives were simultaneously separated within 18 min and 28 min using RP-8 column and RP-18 column, respectively.