44G-15

The first objective was to develop a method to separate the reaction products FB₁-glucose for further characterization and toxicology studies. The second objective was to characterize the reaction kinetics and the products.

The reaction products of FB₁-glucose reaction were separated from excess D-glucose with a reversed-phase C₁₈ cartridge. Mass spectrometry was used to characterize the reaction products. FB₁ was quantified by reversed-phase HPLC with fluorescence detection using preformed o-phthaldialdehyde (OPA) derivatives. FB₁ and FB₁-glucose were determined with an enzyme-linked immunosorbent assay (ELISA).

The results show that reversed-phase C₁₈ chromatography separates FB₁-glucose from excess D-glucose and free FB₁. Mass spectrometry revealed that there were four primary products, N-methyl-FB₁, N-carboxymethyl-FB₁, N-(3-hydroxyacetonyl)-FB₁, and N-(2-hydroxy, 2-carboxyethyl)-FB₁, when FB₁ was heated with D-glucose at 65° C for 48 h. The formation of the FB₁-glucose Schiff’s base was confirmed by mass spectrometry when FB₁ was heated with D-glucose at 60° C. The nonenzymatic browning (NEB) reaction of FB₁ with excess D-glucose followed apparent first order kinetics with respect of FB₁. The activation energy Ea of the reaction was 105.7 kJ mol^-1. At higher temperatures, more complex NEB products were formed. FB₁-contaminated corn could undergo NEB reaction with 0.1 M D-glucose at 60 and 80° C.

The results show that FB₁ can undergo NEB reaction with D-glucose to block FB₁’s primary amine group. The NEB reaction of FB₁ with D-glucose may be used as a process for treating FB₁-containing corn.