A series of glycosphingolipids containing galactose was purified from the earthworm, Pheretima hilgendorfi by DEAE-and QAE-Sephadex, Florisil, and Iatrobeads column chromatography. From structural studies including compositional sugar analysis, methylation analysis, chromium trioxide oxidation, enzymatic hydrolysis, 1H-and 13C-nuclear magnetic resonance spectroscopy, and fast atom bombardment mass spectrometry, their structures were determined to be as follows : Gal β1-Cer, Gal β1-6 Gal β1-Cer, and Gal β1-6 Gal β1-6 Gal β1-Cer.
The ceramide moiety was composed of octadecasphingenine and its branched homologue as the main sphingoids, and saturated C22-, C23-, and C24-acid as the major fatty acids.
Thin-layer chromatography and the results of sugar analysis indicated the structures of two minor components of the ceramide mono-and trihexoside fractions to be Glc-Cer and Man-Gal-Gal-Cer, respectively.