Liquid Chromatography Method for Simultaneous Quantification of ATP and its Degradation Products Compatible with both UV-Vis and Mass Spectrometry
dc.contributor.author | Law, Andrew S. | |
dc.contributor.author | Hafen, Paul S. | |
dc.contributor.author | Brault, Jeffrey J. | |
dc.contributor.department | Anatomy, Cell Biology and Physiology, School of Medicine | |
dc.date.accessioned | 2024-02-22T13:16:44Z | |
dc.date.available | 2024-02-22T13:16:44Z | |
dc.date.issued | 2022 | |
dc.description.abstract | ATP and its degradation products are essential metabolic and signaling molecules. Traditionally, they have been quantified via high-performance liquid chromatography (HPLC) with UV-Vis detection while utilizing phosphate buffer mobile phase, but this approach is incompatible with modern mass detection. The goal of this study was to develop an ultra-performance liquid chromatography (UPLC) method free of phosphate buffer, to allow for analysis of adenine nucleotides with UV-Vis and mass spectrometry (MS) simultaneously. The final conditions used an Acquity HSS T3 premier column with a volatile ammonium acetate buffer to successfully separate and quantify ATP-related analytes in a standard mixture and in extracts from non-contracted and contracted mouse hindlimb muscles. Baseline resolution was achieved with all 10 metabolites, and a lower limit of quantification down to 1 pmol per inject was observed for most metabolites using UV-Vis. Therefore, this method allows for the reliable quantification of adenine nucleotides and their degradation products via UV-Vis and their confirmation and/or identification of unknown peaks via MS. | |
dc.eprint.version | Author's manuscript | |
dc.identifier.citation | Law AS, Hafen PS, Brault JJ. Liquid chromatography method for simultaneous quantification of ATP and its degradation products compatible with both UV-Vis and mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2022;1206:123351. doi:10.1016/j.jchromb.2022.123351 | |
dc.identifier.uri | https://hdl.handle.net/1805/38608 | |
dc.language.iso | en_US | |
dc.publisher | Elsevier | |
dc.relation.isversionof | 10.1016/j.jchromb.2022.123351 | |
dc.relation.journal | Journal of Chromatography B | |
dc.rights | Publisher Policy | |
dc.source | PMC | |
dc.subject | ATP | |
dc.subject | Adenine nucleotides | |
dc.subject | Mouse | |
dc.subject | Muscle | |
dc.subject | UPLC/MS | |
dc.title | Liquid Chromatography Method for Simultaneous Quantification of ATP and its Degradation Products Compatible with both UV-Vis and Mass Spectrometry | |
dc.type | Article |