Colorimetric determination of substances containing the grouping —CH2.CO— in urine extracts as an indication of androgen content

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Abstract
A descr. is given of a modified and improved method of determining androsterone and other 17-ketosteroids by the color reaction with [image]-dinitrobenzene and potassium hydroxide in alcoholic solution. The sources of error and the factors influencing this determination have been investigated. Examination of the behavior of other methyleneketones has shown that the absorption spectrum of the colored solution produced and the rapidity of development of the color vary according to the structure of the compound. The neutral fractions of extracts of hydrolysed urine give the color reaction characteristic of 17-ketosteroids, although with some indication of the presence of other chromogenic compounds. Further, biological activity is associated with the ketonic reaction. These facts justify the use of the reaction for estimating the excretory transformation products of male hormones. A comparison of the chemical with the biological method of determining the excretory products by their activity in the capon comb-growth test has been made with 59 urines from a variety of sources. A significant degree of correlation is found between the colorimetric assay expressed in chromogenic equivalents of androsterone ("sterone") per litre of urine and the biological assay expressed in international units of male hormone activity per litre. It is concluded that colorimetric assay can replace biological assay provided that due regard is paid to the occasional presence of interfering substances, which may be revealed when the approximate absorption spectrum is examined.