10 Amylose = [Amylose] x V x FP x 100
Sample weight titration stage, sample solution was diluted into 50 ml, and then titrated with 0.02 N standardized HCl
until grey color appears. Volume of HCl for titration was then reported. N =
e. Carbohydrate Content by difference AOAC 1995
Carbohydrate = 100 - protein + water + ash + fat
f. Amylose Analysis Apriyanto et al. 1989
First step of amylose analysis was to determine standard curve. Amylose standard curve was made by dissolving 40 mg pure amylose into 1 ml 95 ethanol and 9 ml NaOH 1 N solution. The
mixture was boiled for 10 minutes until gelatinized, cooled, and moved to volumetric flask. Then prepared series mixture in the test tube: 0, 0.2, 0.4, 0.6, 0.8 and 1 ml previous solution were diluted
until 1 ml then add 1 ml acetic acid 1 N and 2 ml Iodine solution 0.2 g iodine and 2 g KI were diluted into 100 ml distillated water. The mixtures were incubated for 20 minutes. Spectrophotometer
Spectronic 21 was used to measure the absorbance of the sample at 625 nm and the data was plotted to make a standard curve.
Amylose content was determined by dissolving 100 mg sweet potato starch into 1 ml ethanol 95 and 9 ml NaOH 1 N solution. Sample solution then boiled for 10 minutes and cooled. Put 5 ml
sample solution into 100 ml volumetric flask and add 1 ml acetic acid 1 N and 2 ml Iodine solution. The mixture was incubated for 20 minutes. Measured the absorbance of the sample at 625 nm and the
absorbance was compared to the standard curve to determined amylose content. Amylose content was determined by:
2. Production of Type 3 Resistant Starch a. Production of Type 3 Resistant Starch Vatanasuchart et al. 2010
Weigh 20 g sweet potato starch in erlenmeyer and add distilled water until reach 200 g of total weight to obtain 10 starch suspension ww. Adjust the acidity of starch suspension to pH 5 with
addition of HCl or NaOH solution. Starch suspension then was autoclaved at 121
o
C under 15 psi for 15 minutes. The suspension was cooled to 50
o
C and was debranched by adding 1 ml pullulanase enzyme 34,779 Umg protein. Place the suspension in a shaking water bath at 50
o
C during 24 hours. After that, deactivated enzyme by heating the suspension at 90
o
C for 30 minutes. Slurry starch was stored at 4
o
C for 24 hours to perform retrogradation. Hydrolyzed starch was centrifuged at 4500 rpm for 15 minutes at room temperature and collect the sediment. The sediment then would be dried using
spray dryer. The resulted product was referred as type 3 resistant starch RS3. Protein Content = Total Nitrogen x Conversion Factor 6.25
11
b. Resistant Starch Analysis Goni et al. 1996
