Separation of several main glycolipids into classes and partially into species by HPLC and UV-detection

The separation and the estimation of several main glycolipid classes as well as of some of these classes into main subclasses (or species) by reverse phase and normal phase high performance liquid chromatography and UV detection, is described. Two different gradient elutions onto a C-18 column and onto a silica column, respectively, and an isocratic elution onto a strong cation exchange column were performed, with detection at 206 nm. Separations were achieved within 30 min. in the reverse phase and normal phase modes and within 10 min. in the cation exchange mode. The following glycolipids standard classes were tested: gangliosides, sulfate cerebrosides (sulfatides), digalactosyldiglycerides, galactosyl-cerebroside as well as ceramides (the backbone of sphingoglycolipids), N-palmitoyl-sphingosine (a synthetic ceramide) and a phospholipid, cardiolipin. Some species of digalactosyl-diglycerides and galactosyl-cerebrosides were also separated. Application of the present method on the separation of glycolipids from animal tissues is represented.