METHOD FOR DETECTING CARBOHYDRATE METABOLITES IN BIOLOGICAL TISSUES

摘要

FIELD: medicine.;SUBSTANCE: before the analysis, each piece of a biological tissue is placed in a separate bottle with an air-tight cap. Acetone is poured in the ratio not less than 1:5, kept for 3 days and more. Further a piece of tissue is removed to measure the acetone volume and the biological tissue weight. Then a portion of acetone is sampled in a porcelain crucible. It is followed by enabled acetone evaporation in an exhaust hood with adding the same amount of distilled water. The sample is mixed up and analysed for carbohydrate metabolites with using enzymatic techniques. An additional weight of the tissue is taken, homogenised in 8% trichloracetic acid. The homogenisate is poured out in volumetric centrifuge tubes with marking a liquid level, performing hydrolysis for 30 min with all glycogen hydrolysed to glucose, centrifuged. Then it is neutralised with 10% NaOH to pH=7-8, reducing the liquid level to the mark, centrifuged. The supernatant is analysed for carbohydrate metabolites with using enzymatic techniques.;EFFECT: detection of a number of carbohydrate metabolites in one sample of the biological material, enabled analysis of the objects for a long period following a sampling procedure.;1 ex