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Clinical Chemistry 13: 717-733, 1967;
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Clinical Chemistry, Vol 13, 717-733, Copyright © 1967 by the American Association for Clinical Chemistry

Determination of Neutral Urinary 17-Oxosteroids with Hydroxide of Hyamine 1O-X in the Zimmermann Reaction

Solveig Bjerre 1 and Rose Kita 1

1 Division of Chemistry and Biochemistry, Department of Laboratories, Toronto General Hospital, Toronto, Canada.

A modified method for 17-oxosteroid determination is described. The organic base, hydroxide of Hyamine lO-X,° which is stable in methanol, has replaced the unstable ethanolic potassium hydroxide solution in the Zimmermann reaction. Maximal color development was studied by varying incubation temperature, time, m-dinitrobenzene, and concentration of hydroxide of Hyamine 10-X. An incubation temperature of 37° for 30 min., with 1% m-dinitrobenzene in 1 M Hyamine 10-X hydroxide solution, gave close to maximal color.

The major urinary 17-oxosteroids, dehydroepiandrosterone, androsterone, and etiocholanolone, gave similar chromogenic values. However, C3 and C20 oxosteroids produced some color in the Zimmermann reaction, and thus if present in large amounts, interfere.

The interference in vitro of 35 drugs was tested, and several drugs were found to interfere.

An essentially pigment-free urinary extract was obtained during the washing step, by addition of water to the ethylene dichloride extract containing sodium hydroxide pellets.

Comparative 17-oxosteroid studies of urine specimens from laboratory personnel and from patients were performed, based on absorbance at a single wavelength (520 mµ) and at three wavelengths (Allen correction). Correlation coefficients for the laboratory personnel were .983 for females and .996 for males; for the patients, .974 and .975, respectively.

Submitted on October 19, 1965
Accepted on September 20, 1966






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Copyright © 1967 by the American Association for Clinical Chemistry.