Separation of Citric Acid Cycle Acids from Animal Tissues by High-Performance Liquid Chromatography

Abstract

A method for separation of the acids of the citric acid cycle and related pathways was developed. The acids were separated by high-performance anion-exchange liquid chromatography on a short column of a strong anion-exchange resin (IEX 520AE) which was eluted with 1/30 M potassium phosphate buffer containing 27.5% (v/v) acetonitrile (pH 3.0) at 40°C. Compounds in the column effluent were detected by ultraviolet spectrophotometry at 210nm. This method was used in the analysis of acids of the citric acid cycle of rat brain and liver and of products of the reaction of succinic thiokinase or fumarase with extracts of animal tissues.