Abstract
A process for extracting the unknown factor from pyloric coeca of skipjack was schemed out as shown in Fig. 1. A preliminarily obtained crystalline crude preparation (Fraction I) was divided into two fractions according to the solubility in a mixture of alcohol and ether. Aside from the insoluble fraction (Fraction II) a soluble fraction (Fraction III) which also proved as crystallizable was further fractionated into the fraction (Fraction IV) adsorbed on a cation-exchanger, Amberlite IRA-400. The aforesaid five fractions were examined of their promotive effect on the acid formation of Leuc. citrovorum on the one hand and of their ultraviolet absorption on the other. The crystalline but still crude preparation obtained as Fraction IV seemed to have advanced fairly in purity, strongly activating Leuc. citrovorum even at a concentration of 40mγ/cc. Mutual comparison of ultraviolet absorption spectra of said five fractions has led to a conclusion that no special absorption peak can be assigned to the unknown factor, though absorption in the range of shorter wave lengths was gradually enhanced with the advance of purification (see Fig. 4) Ionoelectrophoretic runs were carried out with Fraction IV, patterns resembling those of CF and folinic acid, respectively, being obtained (Fig. 5).
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