Journal of Okayama Medical Association
Published by Okayama Medical Association

Full-text articles are available 3 years after publication.

Neotetrazolium塩によるSuccinic Dehydrogenase Systemの活性度測定法とその吟味

Oda, Takuzo
Seki, Shuji
Shibata, Takashi
Sakai, Akira
Okazaki, Hiroaki
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Despite the excellence in bringing about reactions, there are still many difficulties and discrepancies in the use of neotetrazolium chloride for measuring the activity of succinic dehydrogenase system of tissue homogenate. Therefore, with the purpose to find out a measuring method simpler and more accurate, the authors performed some experimets, first on the enzymic reactions of tissue homogenate and then after stopping these reactions by using various agents, tested the ease with which reduction products are extracted with various solvents, and finally compared the absorption spectra of these liquid extracts with those of the products reduced chemically by sodium hydrosulfite. As the results, the extraction of the reduction products with acetone-ether (mixed in equal proprtion v/v) after stopping the reaction with 10 per cent formalin solution was found to be the simplest and best method, and moreover, the absorption spectra of extracts of the products obtained by enzymatical or chemical reduction proved to he uniform and the both of which showed the maximum absorption at the wave length of 520 mμ. In addition, the concentrations of sodium succinate and neotetrazolium chloride, the content and kinds of tissues, pH values and concentration of phosphate buffer solutions, the duration of reaction, aerobic or anaerobic conditions, various factors either enhancing or in hibitory to reaction, were all regulated absolutely or relatively; and by drawing absorption curves of the reduction products, these enzymatic reactions were analyzed and scrutinyzed Then the authors established a method which is general, simple and accurate in measuring the enzymatic activities of a small quantity of fresh tissues in contrast with those of hist chemically stained specimens.