Efficient preparation of sialyloligosaccharides from bovine colostrum facilitated by an improved thin-layer chromatographic monitoring system

Abstract

A new thin-layer chromatographic (TLC) system is described for the investigation of bovine milk sialyloligosaccharides (SOSs). A solvent system, acetone/2-propanol/pyridine/water/acetic acid (60:60:10:30:3, by vol), was developed for the clear separation of sialyl-lactose and sialyl-lactosamine including their isomers, 3′-sialyllactose (3′-SL, Neu5Acα2-3Galβ1-4Glc), 6′-sialyllactose (6′-SL, Neu5Acα2-6Galβ1-4Glc), 3′-sialyllactosamine (3′-SLN, Neu5Acα2-3Galβ1-4GlcNAc), 6′-sialyllactosamine (6′-SLN, Neu5Acα2-6Galβ1-4GlcNAc), as well as disialyllactose (DSL; Neu5Acα2-8Neu5Acα2-3Galβ1-4Glc). The system also permitted good separation of fluorescent labelled sialyllactose isomers modified for the matrix assisted laser desorption/ionisation–time of flight–mass spectrometry analysis. The TLC system can be used for the preparation of SOSs from bovine colostrum. By monitoring the performance of each purification step it was possible, within four weeks, to isolate 100 mg of 3′-SL, 20 mg of 6′-SL, 40 mg of 6′-SLN and 20 mg of DSL, each in more than 90% purity (by high performance liquid chromatography), from 1 L of bovine colostrum.