Atlantic cod Gadus morhua ovaries were incubated in vitro with tritiated 17-hydroxypregn-4-ene-3,20-dione (17-P) to determine whether 17,20beta-dihydroxypregn-4-en-3-one (17,20beta-P) or 17,20beta, 21-trihydroxypregn-4-en-3-one (17,20beta,21-P), or both, were more likely to be the steroid responsible for inducing oocyte final maturation (i.e. resumption of meiosis). Only 17,20beta,21-P was produced, in addition to 11-deoxycortisol (17,21-P), which is intermediate between 17-P and 17,20beta,21-P. Also, the 5beta-reduced forms of 17-P, 17,21-P and 17,20beta,21-P were all found. Some sulphation of 21-hydroxylated steroids was demonstrated. The ability of female G. morhua to make 17,20beta,21-P but not 17,20beta-P was confirmed by radioimmunoassay of plasma samples from spawning fish. Although small amounts of 17,20beta-P immunoreactivity were detected in a few plasma samples, this was shown, by thin-layer chromatography, to be mostly due to cross-reaction with other unidentified compounds. The evidence strongly suggests that 17,20beta,21-P is more likely than 17,20beta-P to be the maturation-inducing steroid in G. morhua.