Abstract
THE measurement of the dihydroxycholanic acids found in human bile (chenodeoxycholic acid, 3α,7α-dihydroxycholanic acid and deoxycholic acid, 3α,12α-dihydroxycholanic acid) or of their ester derivatives or conjugates with glycine or taurine has always presented difficulties where the original material to be analysed contains both acids or derivatives of them. Before 1953, the only method available depended on the repeated precipitation of the barium salt of chenodeoxycholic acid. Its value may be gauged from the fact that until this time chenodeoxycholic acid was considered to be a very minor constituent of human bile, a belief shown to be erroneous by Wootton1 in 1953 when he separated methyl cholate from the two dihydroxy methyl esters by a column chromatographic procedure which utilized silica gel as the supporting medium and pentane/hexane and petroleum ether as the developing solvents. The proportions of methyl chenodeoxycholate and methyl deoxycholate in a mixture could be readily estimated by an investigation of the infra-red spectra of solutions in carbon disulphide. By these means, Wootton demonstrated that chenodeoxycholic acid was, in fact, a major constituent of human bile. The method is relatively insensitive compared with later methods of bile acid measurement which have been devised, requiring about a milligram of the mixed dihydroxy compounds to ensure reasonable accuracy of estimation of the proportion of the two esters.
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References
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Osborn, E. C., Ph.D. thesis. Univ. of London (1959).
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OSBORN, E. Measurement of Chenodeoxycholic Acid and Deoxycholic Acid or their Derivatives in a Mixture. Nature 205, 284 (1965). https://doi.org/10.1038/205284a0
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DOI: https://doi.org/10.1038/205284a0
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