Glycogen Estimation
Glycogen was measured by the Anthrone method of Van der
Vies. [11] Glycogen, present in submicroscopic from in tissue,
yield an amorphous product on reacting with KOH. This
produces a green – brown colour with anthrone reagent. Fifty
milligram tissue of liver was homogenized with 5 ml of 5%
TCA and filtered. To one ml of filtrate, one ml of 10N KOH
was added and the mixture was boiled for exactly 60 min.
Excess alkali was neutralized with 0.5 ml glacial acetic acid
and distilled water is added to make a final volume of 10 ml.
One ml of the above mixture was added to two ml of freshly
prepared anthrone reagent (2 mg anthrone/ml of 36N
sulphuric acid), shaken laterally and heated in a boiling water
bath for 10 min. The green-brown colour develops which was
measured calorimetrically at 650 nm against blank, prepared
simultaneously by using one ml of 5% TCA instead of one
ml of tissue filtrate. The optical density was compared with a
set of glucose standard of varying concentration (0.01 mg/ml
to 0.04 mg/ml). Results have been expressed as µg/mg tissue.