A 100mg of defatted and deproteinated SBP sample was added into tubes containing 4.0 mL of pancreatic α-amylase (10 mg/l) and amyloglucosidase (3 U/mL). The contents in thetube were mixed in a vortex mixer and were incubated at 37 1C for 16 h with a constant stirring at 100 rpm in a mini shaking vortex incubator. The tubes were removed from the incubator exactly after 16 h and 4.0 mL of absolute ethanol was added. The tubes were then centrifuged at 1500 rpm for 10 min. The supernatants were discarded and the pellets were suspended in 2.0 mL 50% ethanol and centrifuged again at 1500 rpm for 10 min. The supernatants were discarded and the pellets were suspended in 50% ethanol and centrifuged at 1500 rpm for 10 min. Once the supernatants were discarded the pellets were air dried.
The pellets were further subjected to determine the non- sucrose carbohydrate fraction; a procedure adapted from Asso- ciation of Official Analytical Chemists, Total Dietary fiber analysis. The pellets were treated with phosphate buffer (pH 6; 5.0 mL) followed by an addition of α-amylase. The solution was incubated for 30min at 951C. Contents were cooled to room temperature and the pH was adjusted to 7.5, followed by an addition protease incubated at 601C for 30min. Further, 280.0 mL of 95% ethanol was added immediately and was kept for 60 min at room temperature. The residue was collected in pre-weighed crucibles. One portion was tested for protein at N 6.25 and the other portion was incinerated at 525 1C for 5 h. The total dietary fiber was calculated as follows: