Abstract
WE wish to report the separation by paper chromatography of a number of steroid glucuronosides and sulphates. The composition of the three systems reported and the conjugates used are given in Table 1, together with the R F values noted in a representative experiment. 10–50 gamma amounts of the free acids or their sodium salts in methanol were spotted on 22½-in. lengths of Whatman No. 1 paper and, after equilibration overnight at room temperature in paper-lined glass tanks, the mobile phase was added to the trough and descending chromatography was carried out for periods of 4–8 hr. Following chromatography, the papers were dried ih a current of air at room temperature, and the conjugates were detected by spraying with, or dipping in, 2–4 per cent phosphomolybdic acid in absolute ethanol followed by heating at 90°–100° C. for 5 min. Conjugate 2 was also detected by dipping in 15 per cent aqueous phosphoric acid, followed by heating at 90° C. for 20 min. and examination under ultra-violet light1, or by dipping in a triphenyltetrazolium chloride reagent prepared by combining immediately before use one volume of 2 per cent triphenyltetrazolium chloride in water with four volumes of 10 per cent sodium hydroxide in 60 per cent aqueous methanol.
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References
Neher, R., and Wettstein, A., Helv. Chim. Acta, 34, 2278 (1951).
Schneider, J. J., Lewbart, M. L., Levitan, P., and Lieberman, S., J. Amer. Chem. Soc., 77, 4184 (1955)
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LEWBART, M., SCHNEIDER, J. Paper Chromatography of Steroid Glucuronosides and Sulphates. Nature 176, 1175 (1955). https://doi.org/10.1038/1761175a0
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DOI: https://doi.org/10.1038/1761175a0
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