Abstract
MOST of the reported chromatographic work on the identification of keto-acids has involved the paper chromatography of a derivative, usually the 2,4-dinitrophenylhydrazone. A simple technique has been developed in this Laboratory whereby acidic materials which may contain keto-acids are chromatographed directly on Whatman No. 1 paper using a solvent containing ethyl acetate/acetic acid/water (60 : 25 : 15 v/v). After irrigation and drying, the papers are sprayed with a saturated alcoholic solution of 2,4-dinitrophenylhydrazine, dried with hot air for 5 min., and then sprayed with an approximately 5 per cent solution of potassium hydroxide in 90 per cent ethanol. The 2,4-dinitrophenylhydrazones which have been formed in situ react to give a variety of colours against an almost colourless background. In general, the derivatives of α-dicarbonyl compounds give colours ranging from light blue to dark purple. Derivatives of other carbonyl compounds produce yellow, red or brown spots. The colour given by any one of these derivatives is often distinctive for that compound, and may sometimes serve to differentiate between keto-acids with similar R F values.
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Lawson, G. J., and Hartley, R. D., Biochem. J., 69, 3 P (1958).
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DOBINSON, F. Detection of Keto-Acids. Nature 183, 675 (1959). https://doi.org/10.1038/183675a0
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DOI: https://doi.org/10.1038/183675a0
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