[8] High-pressure liquid chromatography analysis of neutral glycosphingolipids: Perbenzoylated mono-, di-, tri-, and tetraglycosylceramides

Quantitative high-performance liquid chromatography (HPLC) analysis of glycosphingolipids is a useful and convenient procedure provided high-sensitivity on-line detection is possible. Neutral glycosphingolipids do not possess a characteristic chromophore that permits their quantitative UV detection where high sensitivity is required. However, they can be derivatized with benzoyl chloride or benzoic anhydride to form stable per-O,N- or per-O-benzoylated products, respectively. These products can be quantitatively measured by their absorption of UV light at 280 nm or, with higher sensitivity, at 230 nm. This chapter discusses quantitative analysis of per-O, N-benzoylated neutral glycosphingolipids and of per-O-benzoylated neutral glycosphingolipids. Derivatization of neutral glycosphingolipids is also discussed in detail.