Selected ion monitoring of glycospingolipid mixtures. Identification of several blood group type glycolipids in the small intestine of an individual rabbit

Abstract

A novel application of selected ion monitoring was used for a mixture of non-acid glycosphingolipids of one rabbit small intestine. Earlier studies of permethylated and permethylated-reduced (LiAIH4) derivatives of model compounds have revealed a specificity and abundance of saccharide ions (terminal monosaccharide(s), disaccharide, trisaccharide, etc., and all sugars plus fatty acid) and of ceramide fragments that permit a conclusive detection of separate glycolipid species in a mixture. The sample (50-200 micrograms) was evaporated slowly (1-5 degrees C min-1 from 150-350 degrees C) from the direct inlet probe of an MS 902 mass spectrometer (electron ionization). Mass spectra with fragments up to about m/z 200 were collected on-line by a computer system. A successive partial separation was obtained for glycolipids with from one up to seven sugars. The structures of eight different compounds were identified. They all had 16:0, 22:0 and 24:0 2-hydroxy fatty acids and 18:0 trihydroxy base (phytosphingosine) as major ceramide components. The dominating complex glycolipid was a hexaglycosylceramide with a blood group B type of sequence. A blood group A type sequence was found in a second hexaglycosylceramide. In support of this, the native mixture showed blood group A and B activity. An intense peak, m/z 182, collected from methylated derivatives were evidence for a dominating type 2 carbohydrate chain of the core tetrasaccharide.