title: Simultaneous determination of tricarboxylic acid cycle metabolites by high-performance liquid chromatography with ultraviolet detection.
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.
authors.affiliation: 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.
authors.forename: Yevgeniya I
authors.initials: YI
authors.lastname: Shurubor
authors.affiliation: Department of Biochemistry and Molecular Biology, New York Medical College, Valhalla, NY 10595, USA. Electronic address: arthur_cooper@nymc.edu.
authors.forename: Arthur J L
authors.initials: AJ
authors.lastname: Cooper
authors.affiliation: Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia.
authors.forename: Elena P
authors.initials: EP
authors.lastname: Isakova
authors.affiliation: Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia.
authors.forename: Yulia I
authors.initials: YI
authors.lastname: Deryabina
authors.affiliation: Brain and Mind Research Institute, Department of Neurology, Weill Cornell Medical College, New York, NY 10021, USA.
authors.forename: M Flint
authors.initials: MF
authors.lastname: Beal
authors.affiliation: 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.
authors.forename: Boris F
authors.initials: BF
authors.lastname: Krasnikov
mesh_terms: D000818:Animals; D002264:Carboxylic Acids / Q000097:blood* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002952:Citric Acid Cycle; D005260:Female; D051381:Rats; D017208:Rats, Wistar; D013056:Spectrophotometry, Ultraviolet; D014466:Ultraviolet Rays
chemical_list: D002264:Carboxylic Acids
keywords: Energy metabolism; High-performance liquid chromatography; Organic acids; Tricarboxylic acid cycle; Ultraviolet detection