A 5.0 U of the purified amylase was incubated with 1% (w/v)
soluble starch in 20 mM Tris–HCl buffer (pH 7.5) at 40 ◦C and at
different time intervals. Hydrolysis products were subjected to analyzed
by High-performance liquid chromatography (HPLC) with a
Shodex SH1011 column (8 mm × 300 mm; Waters, USA) at 50 ◦C
using a 0.01 N of sulfuric acid as mobile phase at a flow rate of
1.0 ml/min and the products were detected using refractive index
detector (Waters 2414) maintained at 50 ◦C. Authentic chromatographic
grade glucose and maltose were used as standards for
identification of the hydrolysis products in the reaction mixture.