The detection of cerebrosides on thin layer chromatograms with an anthrone spray reagent
article
Free sugars can be detected on paper or thin layer chromatograms in many ways. In particular, ammoniacal silver nitrate, which reacts with reducing sugars, silver metal being deposited as a brownish spot at the location of the sugar on the chromatograms, is used extensively. Alternatively, spray reagents are used to give furfuraldehyde or one of its derivatives, which can be condensed with an aromatic amine or phenol to give characteristically colored compounds. Spray reagents belonging to this class and used for the present study of detecting cerebrosides on thin layer chromatograms by virtue of their covalently linked sugar moieties were diphenylamine p anisidine, orcinol, aniline hydrogen phthalate and anthrone. Most of the spray reagents tested showed a vague color development, but spraying with a 10% (v/v) sulphuric acid solution, followed by 1% (w/v) anthrone in benzene, was found to give the best results. 4 distinct, green doublet bands were obtained for the cerebrosides from human aortae. From the amounts of material applied to several plates, the lower detection limit of the method was estimated to be ca. 5 μg. Other lipids, such as phosphatidic acid, phosphodiglycerides and cephalins, also present in fraction ii, do not interfere. This method can, however, only be used qualitatively, as the fairly rapid fading of the bands after heating makes quantitative color intensity measurements, by densitometry for example, unreliable.
TNO Identifier
227503
Source
Journal of Chromatography, 85(1), pp. 174-176.
Pages
174-176
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