Radio-enzymic assay of oxo steroids by their reduction with the tritiated form of reduced nicotinamide--adenine dinucleotide

A method of analysis is proposed based on the enzyme-catalysed transfer of tritium from [3H]NADH to suitable substrates. Its practicability is demonstrated on the examples of oestrone and progesterone with the respective use of the 3 beta, 17 beta- and 3 alpha, 20 beta-hydroxysteroid oxidoreductase....

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Bibliographic Details
Published in:Biochemical journal 1975-01, Vol.145 (1), p.47-51
Main Authors: Norymberski, J K, Stoddart, N, Wolstenholme, J B
Format: Article
Language:English
Subjects:
Estrone - blood
Female
Humans
Hydroxysteroid Dehydrogenases
Ketosteroids - analysis
Methods
NAD
Oxidation-Reduction
Pregnancy
Progesterone - blood
Tritium
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Summary:A method of analysis is proposed based on the enzyme-catalysed transfer of tritium from [3H]NADH to suitable substrates. Its practicability is demonstrated on the examples of oestrone and progesterone with the respective use of the 3 beta, 17 beta- and 3 alpha, 20 beta-hydroxysteroid oxidoreductase. Specificity is tested by application to the analysis of the plasma of pregnant women and measurement of the 3H/14C ratios on purification of the enzymic reduction products.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj1450047