A. Gornall, M. MacDonald
1953
Citations
0
Influential Citations
10
Citations
Journal
The Journal of biological chemistry
Abstract
Aromatic hydrazines have been used for many years to form derivatives and permit the separation, purification, and identification of carbohydrate compounds containing carbonyl groups. A number of workers have employed p-nitrophenylhydrazine (l-3) and others 2,4-dinitrophenylhydrazine (4-7). These compounds yielded derivatives which formed deeply colored solutions on addition of alkali. This interesting property has been made the basis of several procedures for the determination of pyruvic acid, acetone, and other substances (8-12). In the field of steroid chemistry, Mason et al. (13) found that cortisone forms a well characterized C-3 monohydrazone with 2,4-dinitrophenylhydrasine in the cold. Chromatographic separation of the 2,4-dinitrophenylhydrazones of estrone and equilenin (14) and of certain androgens (15) has been reported. It has been possible to isolate quantitatively and determine gravimetrically the 2,4-dinitrophenylhydrazone of progesterone (16) but not of testosterone (17). Attempts have been made to identify the steroid hormones histochemically with 2,4-dinitrophenylhydrazine (18, 19). Porter and Silber (20) described the reaction with phenylhydrazine in approximately 22 N sulfuric acid as a calorimetric method specific for the estimation of 17,21-dihydroxy-20-ketosteroids. In our hands their procedure was quite satisfactory in the analysis of relatively pure solutions of these compounds, but it presented difficulties when applied to urine extracts. The chief objections were that the reaction mixture usually turned cloudy and the strong acid alone was found to develop color with extracts of certain urines. They had found that 2,4-dinitrophenylhydrazine was less sensitive in their procedure. The following sect’ion describes conditions under which this hydrazine’ offers special advantages in the quantitative, calorimetric estimation of the steroid hormones in biological extracts.