The assay was initiated by the addition of 100 μL of 120 mM sucrose
into the mixture containing 50 μL sugarcane juice and 50μL
of 1 M sodium acetate (pH 7.5) each. The reaction was
stopped at 60 min by boiling the mixture for 3 min. The
concentration of the glucose released was determined using
the 3,5-dinitrosalicylic acid (DNS) method (Miller 1959).