Abstract
Column chromatography (on cellulose, silicic acid, and Florisil) and thin-layer chromatography were employed for the separation and purification of lipid fractions from normal and Gaucher spleens. A new hydrolysis procedure, followed by paper chromatography, was used for identification of sugar moieties. A nonhydrolytic combined colorimetric procedure, with anthrone and orcinol, was used for the estimation of glucose and galactose separately in glycolipids. The limitations of this method were examined. Spleens from two control subjects and three patients with Gaucher's disease have been examined in detail. In all Gaucher spleens, the predominant feature was the massive acccumulation of glucocerebroside; neutral ceramide oligohexoside levels were probably within the normal range, as were other neutral lipids and phospholipids. In one case examined for gangliosides, these were increased twentyfold. One Gaucher spleen, in which others had reported that the stored "cerebroside" contained predominantly lactose as the saccharide moiety, has been examined in detail and it has been established that the stored material was, in fact, glucocerebroside, ceramide lactoside levels not being significantly elevated. In a further nine cases glucose was the major sugar detected in the splenic lipids.
Highlights
Column chromatography and thin-layer chromatography were employed for the separation and purificationof lipid fractionsfrom normal and Gaucher spleens
We report here examination of the splenic lipids from twelve cases of Gaucher’s disease, including that reported by Parke [6] in which storage of ceramide dihexoside instead of glucocerebroside was suggested
All fractions obtained by cellulose chromatography were examined by T L C and it was found that all glycolipids except ganglioside and all the major phospholipids except phosphatidyl serine were eluted exclusively in the lipid fraction, i.e., with the water-poor phase of chloroform-ethanol-water 16:4
Summary
Column chromatography (on cellulose, silicic acid, and Florisil) and thin-layer chromatography were employed for the separation and purificationof lipid fractionsfrom normal and Gaucher spleens. A nonhydrolytic combined colorimetric procedure, with anthrone and orcinol, was used for the estimation of glucose and galactose separately in glycolipids. The limitations of this method were examined. In all Gaucher spleens, the predominant feature was the massive acccumulation of glucocerebroside ; neutral ceramide oligohexoside levels were probably within the normal range, as were other neutral lipids and phospholipids. One Gaucher spleen, in which others had reported that the stored “cerebroside” contained predominantly lactose as the saccharide moiety, has been examined in detail and it has been established that the stored material was, glucocerebroside, ceramide lactoside levels not being significantly elevated. I n a further nine cases glucose was the major sugar detected in the splenic lipids
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