Design of a High Efficiency Liquid Chromatograph Using Specific Detection and its Evaluation for Analysis of Tricarboxylic Acid Cycle (TCA) Intermediates and Related Compounds on a Nano-Equivalent Scale

Abstract

A modern high efficiency LC-apparatus for the analysis of TCA intermediates and related compounds in the range of 1–30 mµ Eq. = nano Equivalent = nano Val is described. Separation is achieved without gradient elution, for the less polar acids (acetic, fumaric, pyruvic, glutaric, β-hydroxybutyric, lactic, succinic and α-keto-glutaric) within 36 minutes, and for the polar acids (α-keto-glutaric, cis-aconitic, malic, citric and isocitric) within 60 minutes. Possible improvements concerning separation efficiency and increase of detector sensitivity are discussed. The results demonstrate that specific detection by adding an indicator to the eluent can very well go with modern high efficiency liquid/liquid partition column chromatography. A specific detector system is very important for biochemists dealing with biological samples.