In Vitro Starch Digestibility
An in vitro starch digestion method that mimics human digestion was performed according to a variation of the methods of Englyst et al.11 and McCleary and Monaghan.12 About 5 g of fettuccini was accurately weighed into 50 ml Falcon tubes, 45 ml of water was added, and the containers were placed in a boiling water bath for 10 min, which is a defined optimum cooking time for such samples (the time when the white central core of uncooked pasta disappears for the first time: AACC method 66–50). The cooked pasta was rinsed with cold water to stop cooking and drained for 2 min. A volume of 30 ml of water and 0.8 ml of 1 M HCl was added to the sample to attain pH 2.5 (±0.2), 1 ml of 10% pepsin dissolved in 0.05 M HCl was added, and the mixture was stirred slowly (130 rpm) at 15 s intervals. After digesting for 30 min at 37 °C, 2 ml of 1 M NaHCO3 solution was added, and the digestions were adjusted to pH 6.5. Pancreatic digestion commenced with addition of 5 ml 5% pancreatin solution. The contents were immediately adjusted to a volume of 55 ml with distilled water and maintained at 37 °C with gentle stirring. Aliquots of digesta (1 ml) were removed at 20 and 120 min, each added to 4 ml absolute ethanol in 10 ml tubes, and immediately mixed thoroughly. A volume of 0.25 ml of 1% amyloglucosidase was then added, and after incubating for 10 min, 3 ml glucose oxidase-peroxidase reagent was added for measurement of free glucose. Starch measured in the 20 min supernatant was rapidly digested starch (RDS) and starch measured at 120 min was RDS + slowly digested starch (SDS). Resistant starch, starch remaining undigested after 120 min, was also measured. Mean values were calculated from measurements of three batch replicates.
In Vitro Starch Digestibility An in vitro starch digestion method that mimics human digestion was performed according to a variation of the methods of Englyst et al.11 and McCleary and Monaghan.12 About 5 g of fettuccini was accurately weighed into 50 ml Falcon tubes, 45 ml of water was added, and the containers were placed in a boiling water bath for 10 min, which is a defined optimum cooking time for such samples (the time when the white central core of uncooked pasta disappears for the first time: AACC method 66–50). The cooked pasta was rinsed with cold water to stop cooking and drained for 2 min. A volume of 30 ml of water and 0.8 ml of 1 M HCl was added to the sample to attain pH 2.5 (±0.2), 1 ml of 10% pepsin dissolved in 0.05 M HCl was added, and the mixture was stirred slowly (130 rpm) at 15 s intervals. After digesting for 30 min at 37 °C, 2 ml of 1 M NaHCO3 solution was added, and the digestions were adjusted to pH 6.5. Pancreatic digestion commenced with addition of 5 ml 5% pancreatin solution. The contents were immediately adjusted to a volume of 55 ml with distilled water and maintained at 37 °C with gentle stirring. Aliquots of digesta (1 ml) were removed at 20 and 120 min, each added to 4 ml absolute ethanol in 10 ml tubes, and immediately mixed thoroughly. A volume of 0.25 ml of 1% amyloglucosidase was then added, and after incubating for 10 min, 3 ml glucose oxidase-peroxidase reagent was added for measurement of free glucose. Starch measured in the 20 min supernatant was rapidly digested starch (RDS) and starch measured at 120 min was RDS + slowly digested starch (SDS). Resistant starch, starch remaining undigested after 120 min, was also measured. Mean values were calculated from measurements of three batch replicates.
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