Simultaneous determination of tricarboxylic acid cycle metabolites by high-performance liquid chromatography with ultraviolet detection

Yevgeniya I Shurubor; Arthur J L Cooper; Elena P Isakova; Yulia I Deryabina; M Flint Beal; Boris F Krasnikov

doi:10.1016/j.ab.2016.03.004

Simultaneous determination of tricarboxylic acid cycle metabolites by high-performance liquid chromatography with ultraviolet detection

Anal Biochem. 2016 Jun 15:503:8-10. doi: 10.1016/j.ab.2016.03.004. Epub 2016 Mar 19.

Authors

Yevgeniya I Shurubor¹, Arthur J L Cooper², Elena P Isakova³, Yulia I Deryabina³, M Flint Beal⁴, Boris F Krasnikov⁵

Affiliations

¹ Brain and Mind Research Institute, Department of Neurology, Weill Cornell Medical College, New York, NY 10021, USA; Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia.
² Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA. Electronic address: arthur_cooper@nymc.edu.
³ Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia.
⁴ Brain and Mind Research Institute, Department of Neurology, Weill Cornell Medical College, New York, NY 10021, USA.
⁵ Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia; Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA.

PMID: 27001310
DOI: 10.1016/j.ab.2016.03.004

Abstract

Here we describe a simple high-performance liquid chromatography (HPLC) procedure for the simultaneous detection and quantitation in standard solutions of 13 important metabolites of cellular energy metabolism, including 9 tricarboxylic acid (TCA) cycle components and 4 additional metabolites. The metabolites are detected by their absorbance at 210 nm. The procedure does not require prior derivatization, and an analysis can be carried out at ambient temperature within 15 min. The significance of the current work is that the current HPLC procedure should motivate the development of simplified TCA cycle enzyme assays, isotopomer analysis, and determination of selected TCA metabolite levels in plasma/tissues.

Keywords: Energy metabolism; High-performance liquid chromatography; Organic acids; Tricarboxylic acid cycle; Ultraviolet detection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Carboxylic Acids / blood*
Carboxylic Acids / metabolism
Chromatography, High Pressure Liquid / methods*
Citric Acid Cycle*
Female
Rats
Rats, Wistar
Spectrophotometry, Ultraviolet
Ultraviolet Rays*

Substances

Carboxylic Acids