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Abstract
A procedure is described for the analysis of estradiol-3-benzoate, testosterone enanthate benzylic acid hydrazone and estradiol dienanthate in oily solution. The multi-step gradient reversed phase ion-pairing HPLC method uses 1-pentane sulfonic acid sodium salt (0.0025 M in 0.1% acetic acid methanolic solution): acetonitrile and water (55 : 30 to 45 : 15 to 0) mobile phase gradient and a 250 × 4.6 mm, 5 micron octadecyl silane column, with 1,2,4,5-tetrachlorobenzene as internal standard. The time required for chromatography is 31 minutes. The method gives relative standard deviations (RSD) of .94% or better for the assay of active ingredients in commercial formulations.