Spectrophotometric Determination of Steroid Oximes

A spectrophotometric procedure for the determination of steroid oximes is described utilizing 11β-hydroxy-iia-methyl-5β-pregnane -3, 20 -dione dioxime (I) and iia-hydroxy-progesterone dioxime (II) as models. The method is based on (a) acid hydrolysis to release hydroxylamine, (b) oxidation of hydroxylamine to nitrous acid with iodine, (c) diazotization of sulfanilic acid with the nitrous acid, (d) coupling of the diazonium compound with N-(i-naphthyl)-ethylenediamine dihydrochloride, and (e) measurement of the azo dye at 550 nm. A stoichiometric yield of hydroxylamine is obtained. Analyses of I and II have given typical recoveries (mean ± S. D.) of 99.5 ± 1.1% and 97.8 + 3.5%, respectively. The method is sensitive to ≥ 0.3 × 10−5 M dioxime.