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Reverse Phase High-Performance Liquid Chromatography of Cerebrosides, Sulfatides and Ceramides: Microanalysis of Homolog Composition Without Hydrolysis and Application to Cerebroside Analysis in Peripheral Nerves of Adrenoleukodystrophy Patients

1980; Wiley; Volume: 34; Issue: 3 Linguagem: Inglês

10.1111/j.1471-4159.1980.tb11199.x

1471-4159

Shoji Yahara, Hugo W. Moser, Edwin H. Kolodny, Yasuo Kishimoto,

Biochemical Acid Research Studies

Abstract: Reverse phase high‐performance liquid chromatographic analyses of nonhydroxy‐and hydroxycerebrosides and sulfatides, as well as ceramides, have been developed. Cerebrosides, ceramides and sulfatides were perbenzoylated then desulfated before chromatographic analysis. These sphingolipid derivatives can be separated according to their fatty‐acid composition by using an ODS 5‐μ, column (reverse phase column containing octadecylsiloxane derivative of silica, of 5‐μm particle size) and solvent systems consisting of various proportions of acetonitrile and methanol. The retention time of an individual homolog appears to be unaffected by the 4‐trans double bond of the sphingosine moiety. These methods allow us to determine fatty‐acid compositions of ceramides, cerebrosides and sulfatides without hydrolysis. Homolog composition of cerebrosides in peripheral nerve from adrenoleukodystrophy patients was investigated by using reverse phase high‐performance liquid chromatography and was found not to be significantly different from control nerves.