New techniques for the investigation of structure and metabolism of sialic acids

Sialic acid analysis in biological material including gangliosides is often confronted with the necessity to determine trace amounts of various N,O-substituted species. Therefore, techniques of high sensitivity and resolution are required, such as capillary gas-liquid chromatography (GLC) and high performance liquid chromatography (HPLC). Both methods in combination with mass spectrometry allow structural analysis of the different neuraminic acid derivatives. Thus, the number of natural sialic acids known so far has increased to more than 30, including non only saturated, but also 2,3-unsaturated and 2,7-anhydro-sialic acids. Furthermore, HPLC has proved to be especially useful for the study of enzyme reactions, as the sialic acids of enzyme assay mixtures in most cases can be analyzed without prior extensive purification or derivatization.