Page 376 - Demo

380 PROCEEDINGS OF INTERNATIONAL SCIENTIFIC CONFERENCE ON APPLIED BIOTECHNOLOGYweigh approximately 0,5 g of homogeneous fresh sample (0,1 g of powder sample) into 15 mL centrifuge tubes. The test sample is incubated in a simulated physiological environment with amyloglucosidase and pancreatic %u03b1-amylase at 37%u00b0C for 16 h. The digestible starch, under the action of these two enzymes, dissolves and breaks down into glucose. The resistant starch is precipitated using ethanol, then dissolved with KOH solution and hydrolyzed by amyloglucosidase into glucose. The resistant starch content is determined based on the glucose produced by color reaction with GOPOD reagent and measuring absorbance at 510 nm.2.5. Experiments- Investigation of sample preparation: hydrolyze activity of enzymes can be affected by a variety of factors, such as temperature, pH, hydrolysis time and concentration. Therefore, this study focuses on testing and selecting optimal conditions for enzyme reactions.+ Investigating enzyme activity: In this study, the combination of Pancreatic %u03b1-amylase and amyloglucosidase was used as the catalytic agent for hydrolysis in the sample preparation. The aim of the survey of the enzyme mixture%u2019s activity was to optimize the performance of the enzyme, thereby reducing analysis time and associated costs. The activity of Pancreatic %u03b1-amylase was constant at 30 CU/mL while the activity of amyloglucosidase was varied at 0.3 to 15 U/mL.+ Investigating the hydrolysis time of digestible starch and resistant starch: The hydrolysis time for digestible starch was examined at 2 h, 4 h, 6 h, 8 h, 12 h, 14 h, 16 h, and 20 h. Other steps followed the AOAC Official Method 2002.02, with the evaluation conducted on a CRM (Certified Reference Material) sample.