Division 3 Principle

The samples are homogenised by milling. The sample is washed with 40 % ethanol to eliminate soluble sugars and soluble products of starch decomposition.

The enzyme thermostable alpha-amylase is added to the suspension. This enzyme breaks down the starch at 100 °C in smaller chains, whether or not the starch is fully in solution.

Large lumps of starch are broken down very slowly. Therefore it is necessary that the samples should be wholly solved or should be present in the form of a suspension containing very small solid parts.

Then the second enzyme amyloglucosidase is added, which hydrolyses the degraded glucose chains into glucose at 60 °C.

After clarification of the liquid whereby the present proteins, fats and residues are discarded after filtration, a clear solution is obtained which may be used for HPLC.

Separation of the present sugars is done by HPLC.