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SECTION II. APPLIED BIOTECHNOLOGY IN THE PHARMACEUTICAL INDUSTRY... 389of large, unbroken starch molecules. Therefore, the next step will hydrolyze both the excess digested starch and resistant starch into sugar, leading to a positive false positive result. On the contrary, in the case of using amyloglucosidase enzyme with activity > 3.0 U/mL, a part of the resistant starch is hydrolyzed into glucose, leading to a recovery of only 84.6%. The hydrolysis rate and efficiency will increase, helping to convert more starch, including the resistant starch that is difficult to break down in the early stage. This causes the resistant starch content to have a negative error.The results indicated that hydrolysis time significantly affects the resistant starch content in the sample. If the time is less than 12 h, the hydrolysis process is incomplete, and enzymes such as alpha-amylase and amyloglucosidase may not have enough time to break down digestible starch into glucose fully. This results in incomplete removal of digestible starch and some other polysaccharides, causing a positive error in the resistant starch content in the sample. Our data demonstrated that if the digestible starch is not fully hydrolyzed in the initial stage, the measured resistant starch content increases due to the unchanged total starch in the sample. However, if the time exceeds 20 h, overhydrolysis may occur, where the enzymes act on resistant starch, partially hydrolyzing it into glucose and thus reducing its content in the sample. Under the given experimental conditions, between 12 and 20 h, The measured resistant starch content matches the published value for the CRM sample, indicating that digestible starch is likely fully hydrolyzed within these time points.Table 3 shows that the initial sample weight significantly affects the outcomes. Removing the supernatant post-hydrolysis and recovering the resistant starch pellets requires a precise technique. The sample weight must be carefully selected to avoid errors due to losses relative to the initial sample amount.
