2.4. Amylase inhibitor activity
Amylase inhibitor activity was determined according to Dephande, Sathe, Salunkhe, and Cornforth (1982). Triplicate samples, weighing 1g, were extracted in 10 ml distilled water for 12 h at 4 °C in a shaker. The extract was centrifuged at 5000g for 10 min. The supernatant was incubated with α-amylase for 15 min at 37 °C, then starch was added to the mixture and this was incubated for a further 3 min at 37 °C. One unit of enzyme activity was defined as that which liberated, from soluble starch, 1 mg of mallose/min at 37 °C and pH 7.0 under specific conditions.