Determination of Resistant Starch (RS). The RS was
determined using enzymatic method of [5] with some
modifications as follow: The samples were deproteinized
with pepsin (0.2 ml, 1 g pepsin/10 ml of KCl-HCl buffer,
pH 1.5) and the starch was hydrolyzed with pancreatic
alpha amylase (1 ml, 40 mg/ml of Tris maleate buffer, 37
℃ for 16 h). The pellet (containing RS) obtained after
centrifugation (15 min, 3000 x g) was washed with
distilled water and dispersed with 4M KOH followed by
stirring for 30 min at room temperature. The contents
(containing alkali solublized starch equivalent to amount
of RS) were treated with 80 µl of amyloglucosidase (5
mg/ml of acetate buffer pH 4.75) and kept in a water bath
at 60℃for 45 min with constant shaking. The contents
were centrifuged (15 min, 3000 x g) and supernatant
(containing glucose obtained from hydrolysis of alkali
solublized RS) collected in a 500 ml volumetric flask.
The amount of glucose was determined using glucose
oxidase-peroxidase reagent and content of RS was
determined