Arginineの定量
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概要
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The method of colorimetric determination of arginine by hypochlorite or hypobromite procedure based on Sakaguchi's reaction has been modified by several authors. In those methods, it was customary to add urea at the maximal colorization so that the dye formed may not be descomposed by further action of hypochlorite or hypobromite. In the new method described in the present paper, sulfosalicylate has been previously added to the test solution, which would eliminate any hypobromite exceeding the amount necessary to dye formation. The color produced was proved to be stabile. 2 ml. of a mixture containing 2.5 % sulfosalicylic acid, 0.0375 % (M/(200)) giycine and 0.05 % oxine is added to 2ml. of the enzymatic test solution. 2ml. of the filtrate or the centrifuged supernatant is pipetted into a 10ml. graduated test tube. While the test tube is cooled in icewater, 1ml. of a 2.5% sodium hydroxide solution and, 15 minutes later, 1 ml. of hypobromite solution (1g of bromine dissolved in 100ml. of 5 % sodium hydroxide) are added. Then, the colored solution is filled within 10 minutes with water to the mark and immediately colorimetrized with Pulfrich photometer (filter S 50). The extinction coefficient of a red solution, when 1 ml. of M/(4000) arginine (=43.5 γ arginine) and 1 ml. of sulfosalicylic acid-glycin-oxine mixture are used, is 0.71. The Beer's law persists within the range of 87 γ to 5.45 γ arginine. Of the reagents used, the concentration of NaOH has been adjusted so that colorization may reach the highest possible level; and the concentration of oxine has been lowered so that the extinction coefficient may become nil when arginine is absent in the test solution. The above method is simple and accurate and can be used for enzymatic tests. With this method, arginic acid and monoacetylarginine both exhibit the same degree of color formation as that shown by arginine solution of a corresponding concentration. However, those of agmatine, hydroxybutyl-guanidine and β-guanidino propionic acid are about 90%, 76% and 85%, respectively. That of glycocyamine is 66%. Influences of ammonium sulfate, urea, creatine, creatinine, giycine, alanine, leucine, tyrosine, tryptophane and histidine oh colorization of arginine were studied by adding 1 ml. of the solutions of these compounds to 1ml. of M/(4000) arginine solution. The limits of concentrations of these compounds, below which the color development would not be interfered with, are as follows : Giycine, alanine, M/(100) ; urea, M/(200 ; ammonium sulfate, tyrosine and creatinine, M/(400) ; leucine, tryptophane and creatine, M/(800) ; and histidine, M/(4000).
- 千葉大学の論文
- 1958-05-28
著者
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