Then, 1 ml of 5% Phenol was added to each
test tube. After some time, 5 ml of concentrated Sulphuric Acid was
added to all the tubes and shaken well. After ten minutes, the contents
in the test tubes were shaken well and kept in water bath at 25-30°C for
20 minutes. The absorbance of each solution in the sample was noted
using spectrophotometer at 490 nm. Standard graph was prepared
using the standards and the total sugars were calculated for each sample
(Sawhney and Singh, 2006).