Use of Porter–Silber and Schiff Reagents as Spot Tests for Steroids applied on Paper and their Application to the Study of Rat Adrenal Lipids

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Abstract

THE formation of phenylhydrazones with an absorption maximum at 410 mµ by the method of Porter and Silber characterizes steroids with a dihydroxyacetone side-chain. These authors recently reported that the reaction also occurred with the aldehydes of corticosterone and of 11-dehydrocorti-costerone and that with the latter compounds the phenylhydrazone developed at a more rapid rate1. The specificity and difference in speed of the reaction are also evident when it is carried out on paper. A bright yellow colour develops when a region containing a minimum of 2 µgm./cm.2 of the chromogen is passed through the Porter–Silber reagent (25 ml. water, 41 ml. conc. sulphuric acid, 34 ml. ethyl alcohol and 43 mgm. phenylhydrazine hydrochloride). The colour appears instantly with C-21 aldehydes and their acetates, and in 1–2 hr. with steroids containing the dihydroxyacetone side-chain and their acetates. It is stable for days provided the paper is not rinsed or warmed, and under these conditions the paper does not char. Corticosterone, 17-hydroxyprogesterone and other steroids tested gave no colour at a concentration of 25 µgm./cm.2.

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References

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    Silber, R. H., and Porter, C. C., in “Methods of Biochemical Analysis”, 4, 139 (Interscience Publishers, Inc., 1957).

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    Beyler, R. E., and Hoffman, F., J. Amer. Chem. Soc., 79, 5297 (1957).

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    Birmingham, M. K., and Kurlents, E., Can. J. Biochem. Physiol., 37, 510 (1959).

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    Ward, P., and Birmingham, M. K. (in preparation).

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BIRMINGHAM, M. Use of Porter–Silber and Schiff Reagents as Spot Tests for Steroids applied on Paper and their Application to the Study of Rat Adrenal Lipids. Nature 184, BA67–BA68 (1959) doi:10.1038/184067a0b

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