Metabolites were isolated from flash-frozen hearts using a modified Bligh-Dyer methanol/chloroform method (40 (link)). Compounds were separated by liquid-chromatography using a Waters Acquity Ultra Performance Liquid Chromatography unit (Waters Ltd.) and analyzed in negative and positive ion mode in a Sciex 5500 triple quadrupole mass spectrometer (AB Sciex). Full details are provided in the Supplemental Methods.
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