Cerebroside benzoylated

We initially demonstrated that brain cerebrosides, galactosyl-ceramides containing hydroxy fatty acids (HFA) and nonhydroxy fatty acids (NFA), could be completely derivatized by reaction with 10% benzoyl chloride at 60°C for 1 hour (2). After removal of excess reagents by partition between hexane and alkaline aqueous methanol, the perbenzoyl derivatives were seen to separate into two completely resolved components (HFA and NFA... 

A quantitative HPLC method for the analysis of sphingolipids as their perbenzoyl derivatives was first developed for ceramides (5). Ceramides can be conveniently derivatized with benzoic anhydride in pyridine (3 hrs at 110°C) and the products formed have been utilized for the quantitative analysis of NFA and HFA ceramides in normal and Farber s disease tissue. Iwamori and Moser also utilized this procedure for the analysis of ceramides in Farber s disease urine (6). More recently Iwamori and Moser (7) established that the ceramide derivatives formed by reaction with benzoyl chloride or benzoic anhydride are analogous to those formed with cerebrosides. They also characterized the behavior... 

Oxidation of myelin surface cerebrosides by galactose oxidase. Fig. 4 shows silica HPLC of a mixture containing benzoylated-non-hydroxy and hydroxycerebroside and benzoylated derivatives of 2,4-dinitrophenylhydrazone of oxidation products from nonhydroxy- and hydroxycerebroside. Standard curves of two 6-dehydro-derivatives were shown in Fig. 5. These standard curves demonstrate that the response of the benzoylated dinitrophenylhydrazones are linear between 0.025 nmol and 0.6 nmol. Since cerebrosides containing 5 nmol can be determined without tailing to these peaks, this method should allow the determination of as little as 0.5% of the oxidation product. The fact that each curve intersects 0 point in both the abscissa and ordinate indicates that even smaller amounts of these compounds can be detected by this technique. 

Applications of h.p.l.c. to benzoyl derivatives of alditols, glycosides, amino-sugars, aldoses, and alditols has been examined the presence of benzoyl groups has the advantage that u.v. detection methods can be used, and indeed a method for the simultaneous determination of picomole levels of gluco- and galacto-cerebrosides involves benzoylation followed by h.p.l.c.Similarly, tritiated mannitol and fucitol were easily separated after O-benzoylation. ... 

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