7.3.5.3 Chemicals and reagents
• Ethanol 78% (diluted, Agrar-Alkohol, Vienna, UN1170, C-K25V)
• Thermostable α-amylase (Megazyme Kit, Total starch)
• Sodium acetate buffer (100 mM, pH 5.0)
• Amyloglucosidase (Megazyme Kit, Total starch)
• GOPOD reagent (Glucose Determination Reagent)
• D-glucose standard solution (1 mg/ml, Megazyme Kit, Total starch)
7.3.5.4 Procedure
Approximately 100 mg ± 0.1 mg of each sample were weighed into glass test tubes
(triple determination) and 0.2 ml of aqueous ethanol (78%) was added to each tube to
wet the sample and aid dispersion. The tubes were stirred on a vortex mixer.
Immediately 3 ml of thermostable α-amylase (diluted 1:30 in 100 mM sodium acetate
buffer, pH 5.0) were added. Then, the tubes were incubated in a bowling water bath
for 6 min (vigorously stirring all 2 min). Accordingly, the tubes were placed in a water
bath at 50 °C, 0.1 ml amyloglucosidase was added and then, the tubes were stirred and
incubated at 50 °C for 30 min. Subsequent, the entire contents of the test tube were
transferred to a 100 ml volumetric flask (tubes were rinsed with distilled water),
adjusted with distilled water and mixed. Then,