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

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• 作者：Yevgeniya I. Shurubor^a ; ^b ; Arthur J.L. Cooper^c ; ^{arthur_cooper@nymc.edu" class="auth_mail" title="E-mail the corresponding author} ; Elena P. Isakova^b ; Yulia I. Deryabina^b ; M. Flint Beal^a ; Boris F. Krasnikov^b ; ^c
• 关键词：Energy metabolism ; High-performance liquid chromatography ; Organic acids ; Ultraviolet detection ; Tricarboxylic acid cycle
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：15 June 2016
• 年：2016
• 卷：503
• 期：Complete
• 页码：8-10
• 全文大小：420 K

文摘

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.