Fluorometric labeling of tetrahydroprogesterones

7-Diethylaminocourmarin-3-carbohydrazide was used to label the ketone group of the tetrahydroprogesterones to form fluorescent derivatives with high sensitivity. The four isomeric 3-hydroxypregnanes separated readily on high-performance, thin-layer chromatography after derivatization. This separation was not possible with the underivatized isomers. Standards and steroids from biologic mixtures were separated and showed similar characteristics. The methods used are described.