High Performance Liquid Chromatography Analysis of Oligosaccharides Formed During β-Galactosidase Action on Lactose

Abstract

High performance liquid chromatography was used to determine the formation of oligosaccharides during 3-galactosidase action on lactose. Lactose solutions (5 and 20%) in .025 M potassium phosphate buffer (pH 6.60) were inoculated with 3-galactosidase at 1 and 2 /lmol orthonitrophenol units/ml from two different microbial sources and incubated at 4C for 24 h and 37C for 4 h. During hydrolysis, formation of oligosaccharides was followed with high performance liquid chromatography, using both resinbased (/l-Spherogel, Altex Scientific) and polar-bonded (Amino Spheri-5, Brownlee Labs) columns.