DETERMINATION OF THE REACTION-PRODUCT OF GLUTARALDEHYDE AND AMINE BASED ON THE BINDING ABILITY OF COOMASSIE BRILLIANT BLUE

Coomassie Brilliant Blue G-250 (CBBG) was bound to the reaction produc ts of glutaraldehyde with various compounds containing amino group. Th e reaction products of acetaldehyde and malonaldehyde with the amines did not bind with CBBG, indicating that the binding was specific for t he product of glutaraldehyde. The dye binding products rapidly formed in alkaline pH. Gel filtration chromatography indicated that a reactio n product of more than 2 kDa has a dye-binding ability. The reaction p roduct of glutaraldehyde with 6-aminohexanoic acid was separated with SP-Sephadex. CBBG was bound to the first eluted fraction from the colu mn with highest affinity. The absorbance at 595 nm by CBBG binding was linearly related to this lyophilized fraction at a concentration of u p to 250 mu g/ml, suggesting its applicability to the determination of the reaction product of glutaraldehyde and amine. The present method should contribute to our understanding of the chemistry of glutaraldeh yde associated with diverse fields, such as enzyme technology and bioc hemistry.