ION-EXCHANGE CHROMATOGRAPHY OF AMINO ACIDS AND PEPTIDES

I. A NEW DETERMINATION METHOD OF BASIC AMINO ACIDS ON A CARBOXYLIC ACID RESIN

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A new chromatographic method was described for the simultaneous
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Fic. 2. Effect of eluent composition on effluent volume required untill each peak emerges from a 0.6×30cm. column (volume of resin, 8.5ml.) at 30°.
The eluent composition is changed in the mixing proportion of 0.2M, pH 6.0 citrate and 0.2M, pH 6.0 phosphate buffer. It is a matter of course that the resin must be previously buffered by each eluent concerned.
Fic. 3. Effect of eluent composition on effluent volume to peak at pH 6.5 and 30°. The citrate and the phosphate buffer mixed in this case are respectively of 0.2M and pH 6.5. Other notes are the same as in Fig. 2.
determination of lysine, histidine and arginine along with ammonia and tryptophan in mixtures of amino acids. The amino acids were separated by elution analysis on a 0.6×30cm. column of Amberlite IRC-50 (XE-64), using a mixture of 2 volumes of citrate buffer and 1 volume of phosphate buffer (0.2M, pH 6.0 in each) as an eluent, and were estimated from the elution curve with recoveries of 100±3 per cent. The procedure is very convenient since a complete resolution of the amino acids is effected with only 50ml. of the single eluent. Effects of temperature and variations in composition and in pH of the eluent were studied on the position of the emergence of each amino acid, the information from which makes it possible to control the resolution at will.
The author wishes to express his hearty thanks to Prof. T. And o for his valuable advice and encouragement throughout this investigation and to Mr. T. Tamura for the construction of the fraction collector. The expenses of the presnt work were aided partly by the Scientific Research Grant given to Prof. T. Ando from the Ministry of Education, to whom the author's thanks are due.