Starch digestion mechanistic information from the time evolution of molecular size distributions

Size-exclusion chromatography [SEC, also termed gel permeation chromatography (GPC)] is used to measure the time evolution of the distributions of molecular size and of branch length as starch is subjected to in vitro digestion, including studying the development of enzyme-resistant starch. The method is applied to maize starches with varying amylose contents; the starches were extruded so as to provide an analogue for processed food. The initial rates of digestion of amylose and amylopectin components were found to be the same for high-amylose starches. A small starch species, not present in the original starting material, was formed during the digestion process; this new species has a slower digestion rate and is probably formed by retrogradation of longer branches of amylose and amylopectin as they are partially or wholly liberated from their parent starch molecule during the digestion process. The data suggest that the well-known connection between high amylose content and resistant starch arises from the greater number of longer branches, which can form the small retrograded species. The method is useful for the purpose of comparisons between different starches undergoing the process of digestion, by observing the changes in their molecular structures, as an adjunct to detailed studies of the enzyme-resistant fraction.