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Establishing its feasibility on more readily available preclinical 7\u2010T scanners would create new opportunities to study metabolism and physiology in murine models of brain disorders. Here, we demonstrate that the apparent forward rate constant (<jats:italic>k<\/jats:italic><jats:sub>f<\/jats:sub>) of creatine kinase (CK) can be accurately quantified using a progressive saturation\u2010transfer approach in the mouse brain at 7\u2009T. We also find that a 20% reduction in respiration of anesthetized mice can lead to 36% increase in <jats:italic>k<\/jats:italic><jats:sub>f<\/jats:sub> attributable to a drop in cellular pH and mitochondrial ATP production. To achieve this, we used a test\u2013retest analysis to assess the reliability and repeatability of <jats:sup>31<\/jats:sup>P\u2010MRS acquisition, analysis, and experimental design protocols. We report that many <jats:sup>31<\/jats:sup>P\u2010containing metabolites can be reliably measured using a localized 3D\u2010ISIS sequence, which showed highest SNR amplitude, SNR consistency, and minimal T<jats:sub>2<\/jats:sub> relaxation signal loss. Our study identifies key physiological factors influencing mouse brain energy homeostasis in\u00a0vivo and provides a methodological basis to guide future studies interested in implementing <jats:sup>31<\/jats:sup>P\u2010MRS on preclinical 7\u2010T scanners.<\/jats:p>","DOI":"10.1002\/nbm.70055","type":"journal-article","created":{"date-parts":[[2025,5,2]],"date-time":"2025-05-02T08:36:33Z","timestamp":1746174993000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["In Vivo Quantification of Creatine Kinase Kinetics in Mouse Brain Using <sup>31<\/sup>P\u2010MRS at 7\u2009T"],"prefix":"10.1002","volume":"38","author":[{"given":"Mohamed","family":"Tachrount","sequence":"first","affiliation":[{"name":"Wellcome Centre for Integrative Neuroimaging, FMRIB, Nuffield Department of Clinical Neurosciences University of Oxford  Oxford 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