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The occurrence and biosynthesis of steroids in the semen of the elasmobranch fish Squalus acanthias has been reported (Simpson, Wright & Gottfried, 1963; Simpson, Wright & Renfrew, 1964). However, no steroids were detectable in the semen of other species (Simpson, Wright & Hunt, 1963). This report suggests that testosterone and 3β,17-dihydroxypregn-5-en-20-one might be present in the semen of the dogfish Scylliorhinus stellaris.

A steroid extract (chloroform: methanol, 1:1) was prepared from the semen (19 g.) of 5 sexually mature animals. Before extraction, oestriol (10 µg.) was added as an internal standard. The extract was processed sequentially by thin-layer chromatography on Silica Gel GF 254 (250µ thickness) in the systems light petroleum: benzene: ethyl acetate (1:1:4 and 1:1:2). The regions corresponding to oestriol (fraction A) and testosterone (fraction B) were eluted. Further purification and resolution of fraction B was undertaken, since this fraction would be expected to contain in addition to