Abstract
Summary
Adrenal venous blood of the dog has been quantitatively analyzed by paper chromatography for steroid components. Fourteen fractions have been isolated; 3 have been identified as 17-hydroxycorticosterone, corticosterone and 11-desoxy-17-hydroxycorticosterone; 7 of the unidentified fractions exhibit chemical reactions characteristic of adrenocorticosteroids.
Addendum. In a subsequent experiment conducted in collaboraton with Dr. Hans Hirschmann a sample containing Fractions 7 and 8 was acetylated and chromatographed on paper 901/2 hours, employing the system propylene glycol-hexane described by Savard. Five distinct zones were found by examining the strip under ultraviolet light. These subfractions (numbered 1-5 in order of their position on the chromatogram, starting at the origin) were eluted with methanol and were examined spectroscopically in this solvent and after reaction with phenylhydrazine-sulfuric acid reagent. Subfractions 1, 2, and 3 absorbed maximally near 240 mμ; subfraction 4, near 235 mμ, while subfraction 5 showed only a shoulder in this region. With phenylhydrazine reagent subfractions 1 and 5 absorbed maximally at 390-400 mμ, whereas subfractions 2, 3, and 4 gave products which did not exhibit absorption peaks in this region. Each subfraction was treated with acetylcholinesterase in glycylglycine buffer at pH 7.5 for 4 hours at 30°C, and assayed for sodium-retaining activity by the method described in an earlier communication (5). All subfractions were inactive except subfraction 4 which possessed high sodium-retaining activity. No claim of homogeneity is made for any of these subfractions, but it would appear that the sodium-retaining activity and the chromogenicity in the phenylhydrazine reagent shown by our previously described Fraction 8 are not due to the same substance.
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