THE DETERMINATION OF GLYCOGEN IN LIVER AND MUSCLE BY USE OF ANTHRONE REAGENT

TLDR

Current glycogen determination methods rely on KOH or TCA extraction followed by alcohol precipitation and copper reduction or anthrone treatment, with alternative protocols using KOH to destroy alkali‑labile carbohydrates before anthrone analysis. The study reports a detailed anthrone‑adapted method and critically evaluates KOH and TCA extraction protocols. The method extracts liver and muscle tissues with 30 % KOH or 5 % TCA, then quantifies glycogen by anthrone reagent. The anthrone method yields highly specific and precise glycogen measurements.

Abstract

In currently used methods for the determination of glycogen, tissue is extracted either by boiling with 30 per cent potassium hydroxide solution (KOH) or by homogenization with trichloroacetic acid solution (TCA). The glycogen is precipitated from the extract by alcohol and determined either by copper reduction after separation by centrifugation, acid hydrolysis, and neutralization (l-3) or by direct treatment of the precipitated glycogen with anthrone reagent (4-6). An alternative procedure is to destroy alkali-labile carbohydrate by boiling with KOH and then determine the glycogen in the alkali-treated mixture with anthrone reagent (5, 7). Previously, we have reported that a method based upon the use of anthrone reagent gave results of a high degree of specificity and precision (8). This method is a modification of procedures for the determination of dextran (9) and biood sugar (10). In this paper we are reporting in detail our anthrone-adapted method, with critical studies of procedures based upon extraction of tissues with 30 per cent KOH and with 5 per cent TCA.