Stoichiometric dye-binding procedure for the determination of the reactive lysine content of soya bean protein

A new analytical method has been developed for the determination of the reactive lysine content of soya bean protein. The method is based on the reaction of the free basic groups of the protein with 1-phenylazo-2 naphthol-6,8 disulphonic acid. With regard to the stoichiometry of the procedure, it has been proved, contrary to earlier reports, that the basic amino acids, histidine, arginine and lysine, each combine with one mole of the dye. After acylation with propionic anhydride lysine alone loses its dye reactivity. The usefulness of the proposed method has been demonstrated by the determination of the reactive lysine content of several untreated, heat-treated and acid-treated soya bean samples. The results show that heat damage of about 5% in reactive lysine content can be measured in 1·5 h with good reproducibility.