title: A UPLC-MS/MS application for profiling of intermediary energy metabolites in microdialysis samples--a method for high-throughput.
abstract: Research within the field of metabolite profiling has already illuminated our understanding of a variety of physiological and pathological processes. Microdialysis has added further refinement to previous models and has allowed the testing of new hypotheses. In the present study, a new ultra-performance liquid chromatography/electrospray-tandem mass spectrometry (UPLC-ESI-MS/MS) method for the simultaneous detection and quantification of intermediary energy metabolites in microdialysates was developed. The targeted metabolites were mainly from the citric acid cycle in combination with pyruvic acid, lactic acid, and the ATP (adenosine triphosphate) hydrolysis product adenosine along with metabolites of adenosine. This method was successfully applied to analyze the microdialysates obtained from an experimental animal study giving insight into the hitherto unknown concentration of many interstitial energy metabolites, such as succinic acid and malic acid. With a total cycle time of 3 min, injection to injection, this method permits analysis of a much larger number of samples in comparison with conventional high performance liquid chromatography/tandem mass spectrometry HPLC-MS/MS strategies. With this novel combination where microdialysis and high sensitivity UPLC-MS/MS technique is combined within cardiologic research, new insights into the intermediary energy metabolism during ischemia-reperfusion is now feasible.
authors.affiliation: Section for Toxicology and Drug Analysis, Department of Forensic Medicine, Aarhus University, Denmark.
authors.forename: Rune Isak Dupont
authors.initials: RI
authors.lastname: Birkler
authors.forename: Nicolaj Brejnholt
authors.initials: NB
authors.lastname: Støttrup
authors.forename: Sigurd
authors.initials: S
authors.lastname: Hermannson
authors.forename: Torsten Toftegaard
authors.initials: TT
authors.lastname: Nielsen
authors.forename: Niels
authors.initials: N
authors.lastname: Gregersen
authors.forename: Hans Erik
authors.initials: HE
authors.lastname: Bøtker
authors.forename: Mette Findal
authors.initials: MF
authors.lastname: Andreasen
authors.forename: Mogens
authors.initials: M
authors.lastname: Johannsen
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D004734:Energy Metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D057230:Limit of Detection; D017551:Microdialysis / Q000379:methods*; D009206:Myocardium / Q000378:metabolism; D013552:Swine; D053719:Tandem Mass Spectrometry / Q000379:methods*