濾紙クロマトグラフによる蛋白質中の酸性アミノ酸の定量について

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The content of the acidic amino-acids has much importance upon the physico-chemical proper-ties of the protein because the number of the anionic groups depends largely on the content of the acidic residues. The present report describes the paper-chromatographic method for determining the acidic amino-acid contents, some critical experiments on its details, and the discussion of the results obtained by the application of this method to some proteins. The hydrolized samples are put on a horizontal starting line on a filter paper, side by side with the known quantities of the acidic amino-acid solutions, and the spots are chromatographed upward simultaneously by aqueous phenol, after dried, the paper is sprayed with ninhydrin solution, the coloured spots being developed by heating the filter paper in 90-100°C. atmosphere saturated with steam. The spots of the acidic amino-acids are cut off and the colour of each spot is eluted out with 50% ethanol and evaluated electrophotometrically at 570mμ. The content in the sample solution is obtained by the aid of the standard curves of the known solutions chromato-graphed on the same falter paper. For coloration of the spots, the a reasonable period of heating was 7.5-10 minutes (Fig. 1); the colour increased linearly with the quantity of the ammo-acids between 10-60μg. per spot (Fig. 2) ; the mean recovery, including from the hydrolysis to the last process of the method, was 96.5% for aspartic acid, and 100.9% for glutamic acid respectively. The values obtained by analyzing gelatin by the present method were in good agreement with the data reported previously (Table 1). However, the values of glutamic acid in the muscle preparations of some marine animals were slightly higher than those of the same preparations obtained with microbiologicalassay by SUGIMURA et al. (Tables 2 & 3).
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濾紙クロマトグラフによる蛋白質中の酸性アミノ酸の定量について

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