A METHOD FOR THE SEPARATION OF TRICARBOXYLIC CYCLE INTERMEDIATES BY CELITE-COLUMN CHROMATOGRAPHY AND ITS APPLICATION TO THE TISSUES OF ASCARIS LUMBRICOIDES VAR. SUIS

Abstract

1. A method of column chromatography was described, in which 0.2 N H₂SO₄ on Celite was used as the stationary phase and n-butanol-chloroform or isoamyl alcohol-chloroform mixtures as the solvent system.
2. Main intermediates of the tricarboxylic acid-cycle were separated by the above mentioned two solvent systems and determined by titration with 0.002 N KOH. The recovery of the acids ranged from 80 to 90 per cent.
3. This method was applied to the determination of the amount of members of the tricarboxylic acid-cycle in tissues of normal rat. The results were in good accord with the previous reports obtained by different procedures.
4. Application of the above mentioned method to the tissues of Ascaris lumbricoides var. suis showed that succinic acid was the main component in muscle as well as perienteric fluid, whereas no significant amount of tri-carboxylic acids were discovered in these tissues. Problems concerning metabolism in ascaris were discussed from this unique feature in distribution of the acids found.
The authors wish to express their thanks to Prof. Y. Kobayashi and to Dr. S. Ebashi for their continued interest in this work, and to Dr. T. Tatsuno for synthe-sizing a-ketoglutaric and oxalacetic acids. The authors were also indebted to Miss K. Hamano for her technical assistance.