The Use of the Sulfuric Acid Reaction for the Estimation of (y- and /?-estradiols and of Estrone and Equilin in Binary Mix- Tures in Pure Solutions*
نویسنده
چکیده
The problem of estimating quantitatively the components of a mixture of naturally occurring estrogens has been attacked through the preparation of their p-phenylazobenzoyl (azoyl) esters and separating these esters by the chromatographic adsorption technique (1). By this method, the monoazoates were quantitatively separated from the diazoates. Hydrolysis of these esters and suitable extraction procedures resulted in the quantitative recovery of the free estrogens. The present report describes a study of a single phase sulfuric acid reaction for the further analysis of the mixtures of free estrogens so obtained and for the positive identification of a single free estrogen. It was early observed that, when the natural estrogens are treated with sulfuric acid, orange-colored solutions possessing a green fluorescence are obtained (2-4). Kober (4) showed that the initial orange color obtained with estrone was changed to a clear green-fluorescing red on warming with water, and, in addition, that the intensity of the red color could be enhanced and the intensity of the yellow color and the fluorescence diminished by using a mixture of phenol and sulfuric acid in the initial stage of the reaction instead of sulfuric acid alone. Since the orange or yellow solutions resulting from the action of sulfuric acid on cholesterol, bile acids, pregnanediol, and many other steroids are decolorized by dilution with water, the test was claimed to be specific for the natural estrogens. Cohen and Marrian (5), however, found that the presence of either cholesterol or pregnanediol considerably decreased the intensity of the red color yielded by given quantities of estrone or estriol in the Kober reaction. They resorted, therefore, to an alkali extraction procedure which separated the
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تاریخ انتشار 2003