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Clinical blood lactate testing is conducted with laboratory analyzers, leading to a delay of 3 h between triage and lactate result. Here, a fluorescence\u2010based blood lactate assay, which can be utilized for bedside testing, based on measuring the hydrogen peroxide generated by the enzymatic oxidation of lactate is described. To establish a hydrogen peroxide assay, near\u2010infrared cyanine derivatives are screened and sulfo\u2010cyanine 7 is identified as a new horseradish peroxidase (HRP) substrate, which loses its fluorescence in presence of HRP and hydrogen peroxide. As hydrogen peroxide is rapidly cleared by erythrocytic catalase and glutathione peroxidase, sulfo\u2010cyanine 7, HRP, and lactate oxidase are encapsulated in a liposomal reaction compartment. In lactate\u2010spiked bovine whole blood, the newly developed lactate assay exhibits a linear response in a clinically relevant range after 10 min. Substituting lactate oxidase with glucose and alcohol oxidase allows for blood glucose, ethanol, and methanol biosensing, respectively. This easy\u2010to\u2010use, rapid, and versatile assay may be useful for the quantification of a variety of enzymatically oxidizable metabolites, drugs, and toxic substances in blood and potentially other biological fluids.<\/jats:p>","DOI":"10.1002\/smll.202000369","type":"journal-article","created":{"date-parts":[[2020,4,24]],"date-time":"2020-04-24T03:27:47Z","timestamp":1587698867000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Near\u2010Infrared Fluorescence Hydrogen Peroxide Assay for Versatile Metabolite Biosensing in Whole Blood"],"prefix":"10.1002","volume":"16","author":[{"given":"Simon","family":"Matoori","sequence":"first","affiliation":[{"name":"John A. Paulson School of Engineering and Applied Sciences Harvard University  Cambridge MA 02138 USA"},{"name":"Wyss Institute of Biologically Inspired Engineering Harvard University  Boston MA 02115 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9406-773X","authenticated-orcid":false,"given":"David J.","family":"Mooney","sequence":"additional","affiliation":[{"name":"John A. 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