Determination of Glycogen in Tissues*
نویسنده
چکیده
The purpose of the present paper is to present some simplifying modifications of Pfltiger’s method (1) for the estimation of glycogen, that render it available for small amounts of tissue. The suggested modifications involve (a) shortening of the time necessary to hydrolyze the tissue with strong alkali, (b) facilitation of the separation of the glycogen by precipitating it in the presence of charcoal and centrifuging, (c) hydrolysis of the separated glycogen for a shorter time than customary, and (d) the substitution of sulfuric acid for hydrochloric acid to avoid the introduction of chlorides. (a) Less than the 2 hours of hydrolysis with strong potassium hydroxide recommended by Pfliiger (1) has been found sufficient by Schiindorff and coworkers (2) who using 30 per cent potassium hydroxide found 30 minutes sufficient for liver and muscle, provided the flasks were shaken thoroughly every 5 to 10 minutes. Bierry and Gruzewska (3) obtained good results by autoclaving at 120” for 30 minutes with 35 per cent potassium hydroxide. The time recommended in the writer’s procedure is 30 to 40 minutes. (b) Pfltiger (4) found the precipitation of glycogen by alcohol from its solutions in strong alkali may take place so slowly that there is danger of losses in its estimation. The procedure recommended in the present paper avoids this danger by adsorbing the glycogen on activated charcoal which is found not to interfere with the subsequent sugar determination. Moreover, when the washed glycogen is hydrolyzed, the filtration and clarification of
منابع مشابه
The determination of glycogen in liver and muscle by use of anthrone reagent.
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 ...
متن کاملThe Determination of Glycogen in Liver and Muscle by Use of Anthrone Reagent* by Nicholas
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 ...
متن کاملPreparation of Tissue Extracts for the Determination of Malic Acid* by Lawrence
Hummel’s (1) fluorometric determination of malic acid requires the precipitation of calcium malate by alcohol as a means of fractionation from biological materials. Fructose-l, 6-diphosphate, glucose-6-phosphate, or glycogen if present must be removed. Below is described a method for preparing extracts of tissues which applies the principles of chromatography on silica gel. Osazone formation, p...
متن کاملThe determination of different effective concentration of ethanolic extract of bee pollen on biochemical analysis in liver, spleen and heart tissues of rainbow trout, Oncorhynchus mykiss (Walbaum, 1792)
The aim of this study was to investigate the effective concentration of ethanolic extract of bee pollen on liver, spleen and heart tissues of fish. Bee pollen extract in various concentrations (0.5, 2.5, 5, 10, 20 and 30 ppm) was administered to fish for 96 h under aquarium conditions. The malondialdehyde (MDA) levels, total antioxidant status (TAS), total oxidant status (TOS), oxidative stres...
متن کاملHomogenization versus homogenization-free method to measure muscle glycogen fractions.
OBJECTIVE The glycogen is extracted from animal tissues with or without homogenization using cold perchloric acid. Three methods were compared for determination of glycogen in rat muscle at different physiological states. MATERIALS AND METHODS Two groups of five rats were kept at rest or 45 minutes muscular activity. The glycogen fractions were extracted and measured by using three methods. ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره شماره
صفحات -
تاریخ انتشار 2003