SPECTROPHOTOMETRIC STUDIES OF THE m-DINITRO BENZENE REACTION WITH SEX STEROIDS1

IN 1935 ZIMMERMANN (I) described a color reaction between m dinitrobenne and several sex hormones. Since that time several modifications of the reaction have been described and it has been used for the quantitative determination of excreted androgens (2, 3, 4). We have investigated these methods (to be reported in another paper) and because of the difficulties and discrepancies which were encountered, we have made an extensive spectrophotometric investigation of the reaction. For the pursuance of the spectrophotometric studies the method of Callow was used, for of all the methods it is the most rigorous in regard to purity of solvent and reagents, regulation of time, temperature and light and most important, it gives consistently reproducible results with crystalline hormones. The steroids2 were dissolved in absolute alcohol in such concentration that themaximum amount of substance used was contained in 0.2 cc. of solution