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Clinical Chemistry 11: 554-562, 1965;
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Clinical Chemistry, Vol 11, 554-562, Copyright © 1965 by the American Association for Clinical Chemistry

Relative Rates of Color Development of 17-Keto- and Ketogenic Steroids in the Zimmermann Reaction

John H.C. Liu 1

1 Trainee, 1962-3, Postgraduate Training Program for Steroid Biochemistry, National Institutes of Health, U. S. Public Health Service, Clark University, Worcester 10, Mass. Present address: 2909 Hillcroft, Houston 27, Tex.

The rates of reaction at 27° 37°, 47°, and 57°, of 17-ketosteroids (17-KS), dehydroisoandrosterone (DHA), and 17-ketogenic steroids (17-KGS) were studied by a modified Zimmermann color reaction. Curves were constructed by plotting absorbance against time necessary for maximum color development at these temperatures. Pooled eluates of 17-KS and 17-KGS from normal urine samples were used in the determinations. Temperature was plotted against the time found necessary for maximum color development using DHA as standard. Experimental results conformed to the temperature-dependence theory of reaction rates, that for every 10° rise, the rate of reaction doubles. Curves of the absorbances at various times of incubation were calculated as percentiles of maximum color formation at each temperature. The interrelationship between 17-KS, 17-KGS, and DHA with respect to optimum reading time was noted. The significance of the finding that the rate of reaction of 17-KGS is much greater than that of 17-KS and DHA is demonstrated. Functional group contribution to the rate of reaction is discussed, as are other factors affecting the Zimmermann color reaction.

Submitted on August 10, 1962






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Copyright © 1965 by the American Association for Clinical Chemistry.