Abstract
This report describes a radioimmunoassay method for 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-diOHP) in the blood plasma of Atlantic salmon ( Salmo salar). 3H-labeled 17α,20β-diOHP was synthesized from 17α-[ 3H]hydroxy-4-pregnen-3-one (17α-OHP) by enzymatic reduction of the C-20 oxo group and the simultaneous oxidation of coenzyme-reduced nicotinamide adenine dinucleotide. Antiserum to 17α,20β-diOHP-3-CMO-BSA exhibited high specificity; all steroids tested against the antiserum gave less than 1% cross-reactivity at 50% displacement. Crude dichloromethane extracts of small volumes (50 μl or less) of plasma can be assayed directly without prior purification. The working range is 5 to 300 pg. Interassay and intraassay coefficients of variation were less than 10%. Various parallelism tests showed high accuracy and sensitivity. The assay is simple and rapid and at least 75 samples can be easily assayed in duplicate by two analysts in 1 day.
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