ВЪРХУ ЕДИН МЕТОД ЗА КАЧЕСТВЕНО ДОКАЗВАНЕ НА ДИЗАХАРИДАЗНА АКТИВНОСТ В ТЪНКОЧРЕВНА ЛИГАВИЦА

Abstract

Description of a method for qualitative demonstration of disaccharidase activity in the small intestinal mucosa is presented, which allows bbe carrying out o f semiquantitative assessment also. A piece of the mucosa, measuring 4-5 mm (abou 25 mm2 ) is incubated in a humid chamber with 6.15 ml substrate solution at 37° C for a 45 min period. The piece is placed over a sterile object glass with a depression wttnein the substrate is dripped. 5 % saccharose solution, 5 % trehalose and 10 % solution of lactose in mallein buffer at ph 5.8 are used as substrate. After the incubation time is over, the mucosal piece is removed and the glucose obtained as a result of hydrolysis is specifically demonstrated with the glucose-oxidase test papers Biophan G.The colour appearing at the reagent end of the indicator papers 2 min after ist humidization in the substrate solution is checked with the colour scale attached to the test and on the basis of the latter.the result is recorded with 0.+ up to ++++. The comparative study of 68 samples from small intestinal mucosa of rats showed that + usually corresponds to activity between 1 and 2 quantitative units (U= micro-Mol/min hydrolized substrate for 1 g fresh mucosa); ++ corresponds to activity from 2U to 4U; +++ and ++++ corresponds to disaccharidase activity exceeding 4.6 U under experimental conditions. The execution of the method is simple and the results are available within 1hour which renders it particularly convenient fo r mass, large-scale examinations. Results are reported of investigating 336 samples from small intestines of rats an 72 samples from human small intestinal mucosa. The weaker lactase activity as compared to saccharase and trehalase activity is emphasised. Some theoretical basic points, cosidered as prerequites for the accomplishement of the method, are discussed.