NONENZYMIC SULFATION OF ANDROSTERONE WITH L-ASCORBIC ACID 3-SULFATE

To test the hypothesis that the purpose of ascorbic acid in steroidogenic organs is to serve as a sulfating agent, either in conjunction with or independent of PAPS, L-ascorbic acid 3-sulfate was synthesized by a three step procedure: (1) preparation of 5, 6-0-benzylidene ascorbic acid from L-ascorbic acid and benzaldehyde; (2) synthesis of 5,6-0-benzylidene ascorbic acid 3-sulfate with pyridine-sulfur trioxide in N,N-dimethylformamide; and (3) removal of the benzylidene group by mild acidic hydrolysis. Evidence is presented that sulfation of androsterone (3α-hydroxy-5α-androstan-17-one) with ascorbic acid 3-sulfate produces androsterone sulfate. Identification of androsterone sulfate was accomplished by chromatography, electrophoresis, countercurrent distribution, infrared spectrophotometry and radiochemical analysis as well as by the classical tests for steroid sulfate. This experiment demonstrates that ascorbic acid 3-sulfate can serve as a sulfating agent under the proper conditions. © 1968.