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December 1955

Corticosteroids in the Aqueous Humor of the RabbitPaper Chromatography

Author Affiliations

Department of Research, Wills Eye Hospital.

AMA Arch Ophthalmol. 1955;54(6):853-857. doi:10.1001/archopht.1955.00930020859007

In a previous investigation1 analysis of chloroform extracts of aqueous humor from normal rabbit eyes by the blue tetrazolium method indicated the presence of corticosteroids in quantities of the order of magnitude of 1-1.5γ per milliliter. Since the blue tetrazolium method neither permits the identification of individual compounds in a mixture nor is specific only for oxidizable corticoids, the extracted aqueous humor was fractionated by paper partition chromatography and analyzed for corticosteroids. The results of this investigation strongly indicate that at least part of the blue-tetrazoliumpositive material present in aqueous humor is hydrocortisone.

MATERIALS  The following purified crystalline corticosteroids were donated by Sharp & Dohme, Division of Merck & Company, Inc.: 11-dehydrocorticosterone (A); corticosterone (B); 11-desoxy-17-hydroxycorticosterone (S); cortisone (11-dehydro-17hydroxycorticosterone, Compound E); hydrocortisone (17-hydroxycorticosterone, Compound F); hydrocortisone acetate; dihydrocortisone, and tetrahydrocortisone. Small aliquots of the samples were dissolved in absolute methanol in concentrations of 1γ and 37 per microliter and

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