{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T21:47:44Z","timestamp":1789422464946,"version":"build-2803163510"},"reference-count":33,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,3,4]],"date-time":"2023-03-04T00:00:00Z","timestamp":1677888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Chongqing Postdoctoral Science Foundation, China","award":["CSTB2022NSCQ-BHX0687"],"award-info":[{"award-number":["CSTB2022NSCQ-BHX0687"]}]},{"name":"Chongqing Postdoctoral Science Foundation, China","award":["2023MSXM006"],"award-info":[{"award-number":["2023MSXM006"]}]},{"name":"Chongqing Postdoctoral Science Foundation, China","award":["YGZ2021133"],"award-info":[{"award-number":["YGZ2021133"]}]},{"name":"Chongqing Medical Scientific Research Project (Joint project of Chongqing Health Commission and Science and Technology Bureau), China","award":["CSTB2022NSCQ-BHX0687"],"award-info":[{"award-number":["CSTB2022NSCQ-BHX0687"]}]},{"name":"Chongqing Medical Scientific Research Project (Joint project of Chongqing Health Commission and Science and Technology Bureau), China","award":["2023MSXM006"],"award-info":[{"award-number":["2023MSXM006"]}]},{"name":"Chongqing Medical Scientific Research Project (Joint project of Chongqing Health Commission and Science and Technology Bureau), China","award":["YGZ2021133"],"award-info":[{"award-number":["YGZ2021133"]}]},{"name":"Chongqing Medical and Pharmaceutical College, China","award":["CSTB2022NSCQ-BHX0687"],"award-info":[{"award-number":["CSTB2022NSCQ-BHX0687"]}]},{"name":"Chongqing Medical and Pharmaceutical College, China","award":["2023MSXM006"],"award-info":[{"award-number":["2023MSXM006"]}]},{"name":"Chongqing Medical and Pharmaceutical College, China","award":["YGZ2021133"],"award-info":[{"award-number":["YGZ2021133"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>In this study, a copper hexacyanoferrate nanoparticle with excellent oxidase-mimetic behaviour has been synthesized through a simple precipitation method. The synthesized copper hexacyanoferrate nanoparticle has intrinsic oxidase-like activity, which can catalyze the chromogenic reaction of 2,2\u2032-azinobis-(3-ethylbenzthiazoline-6-sulphonate) through an O2\u2022\u2212 reactive oxygen-species-participated process. On the other hand, K3[Fe(CN)6] can be reduced by ascorbic acid (AA) to produce K4[Fe(CN)6], thereby inhibiting the formation of the copper hexacyanoferrate nanoparticles. Furthermore, ascorbate oxidase (AAO) can catalyze the oxidation of AA to produce dehydroascorbic acid, which cannot reduce K3[Fe(CN)6]. Thus, a system for an AAO-regulated in situ formation of copper hexacyanoferrate nanoparticles was constructed by coupling a prepared copper hexacyanoferrate nanozyme with AA for the detection of AAO activity. This colorimetric sensing assay shows high sensitivity and selectivity for the detection of AAO activity (the limit of detection is 0.52 U\/L) with a linear range of 1.1\u201335.7 U\/L. Finally, the developed method was applied to detect the activity of AAO in normal human serum with a satisfactory sample spiked recovery (87.4\u2013108.8%). In short, this study provides a good strategy for the construction of nanozyme-based multi-enzyme cascade-signal amplification assay.<\/jats:p>","DOI":"10.3390\/bios13030344","type":"journal-article","created":{"date-parts":[[2023,3,6]],"date-time":"2023-03-06T02:28:34Z","timestamp":1678069714000},"page":"344","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Enzyme-Regulated In Situ Formation of Copper Hexacyanoferrate Nanoparticles with Oxidase-Mimetic Behaviour for Colorimetric Detection of Ascorbate Oxidase"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2229-9125","authenticated-orcid":false,"given":"Hao","family":"Zhang","sequence":"first","affiliation":[{"name":"Chongqing Key Laboratory of High Active Traditional Chinese Drug Delivery System, Chongqing Medical and Pharmaceutical College, Chongqing 401331, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dan-Ni","family":"Yang","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of High Active Traditional Chinese Drug Delivery System, Chongqing Medical and Pharmaceutical College, Chongqing 401331, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Li","sequence":"additional","affiliation":[{"name":"Chongqing Key Laboratory of High Active Traditional Chinese Drug Delivery System, Chongqing Medical and Pharmaceutical College, Chongqing 401331, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9300-5710","authenticated-orcid":false,"given":"Feng-Qing","family":"Yang","sequence":"additional","affiliation":[{"name":"School of Chemistry and Chemical Engineering, Chongqing University, Chongqing 401331, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1016\/j.snb.2018.11.020","article-title":"Non-oxidation reduction strategy for highly selective detection of ascorbic acid with dualratio fluorescence and colorimetric signals","volume":"281","author":"Chen","year":"2019","journal-title":"Sens. Actuator B-Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.aca.2019.09.054","article-title":"A ratiometric fluorescence-scattered light strategy based on MoS2 quantum dots\/CoOOH nanoflakes system for ascorbic acid detection","volume":"1091","author":"Wu","year":"2019","journal-title":"Anal. Chim. Acta"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"15477","DOI":"10.1021\/acs.analchem.9b03272","article-title":"Carbon dots as fluorescent\/colorimetric probes for real-time detection of hypochlorite and ascorbic acid in cells and body fluid","volume":"91","author":"Wei","year":"2019","journal-title":"Anal. Chem."},{"key":"ref_4","unstructured":"Institute of Medicine (2001). Dietary Reference Intakes for Vitamin A, Vitamin K, Arsenic, Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybdenum, Nickel, Silicon, Vanadium, and Zinc, The National Academies Press."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"147269","DOI":"10.1016\/j.apsusc.2020.147269","article-title":"Exploration of pH-responsive carbon dots for detecting nitrite and ascorbic acid","volume":"530","author":"Gan","year":"2020","journal-title":"Appl. Surf. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1016\/j.jmst.2020.07.037","article-title":"2D-titanium carbide (MXene) based selective electrochemical sensor for simultaneous detection of ascorbic acid, dopamine and uric acid","volume":"72","author":"Murugan","year":"2021","journal-title":"J. Mater. Sci. Technol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1016\/j.jpba.2007.10.033","article-title":"FIA determination of ascorbic acid at low potential using a ruthenium oxide hexacyanoferrate modified carbon electrode","volume":"46","author":"Bertotti","year":"2008","journal-title":"J. Pharm. Biomed. Anal."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"11795","DOI":"10.1021\/ja2045925","article-title":"Hyperpolarized [1-13C]-ascorbic and dehydroascorbic acid: Vitamin C as a probe for imaging redox status in vivo","volume":"133","author":"Bohndiek","year":"2011","journal-title":"J. Am. Chem. Soc."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.jplph.2005.09.016","article-title":"Ascorbic acid oxidase is dynamically regulated by light and oxygen. A tool for oxygen management in plants?","volume":"164","author":"Ciraci","year":"2007","journal-title":"J. Plant Physiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"121522","DOI":"10.1016\/j.talanta.2020.121522","article-title":"Redox reaction-modulated fluorescence biosensor for ascorbic acid oxidase assay by using MoS2 quantum dots as fluorescence probe","volume":"222","author":"Li","year":"2021","journal-title":"Talanta"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"129910","DOI":"10.1016\/j.snb.2021.129910","article-title":"Self-assembled dual-emissive nanoprobe with metal-organic frameworks as scaffolds for enhanced ascorbic acid and ascorbate oxidase sensing","volume":"339","author":"Chen","year":"2021","journal-title":"Sens. Actuator B-Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1007\/s00604-018-2954-8","article-title":"A dual-model strategy for fluorometric determination of ascorbic acid and of ascorbic acid oxidase activity by using DNA-templated gold-silver nanoclusters","volume":"185","author":"Liu","year":"2018","journal-title":"Microchim. Acta"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1007\/s00604-019-3341-9","article-title":"A carbon dot-based ratiometric fluorometric and colorimetric method for determination of ascorbic acid and of the activity of ascorbic acid oxidase","volume":"186","author":"Wang","year":"2019","journal-title":"Microchim. Acta"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1038\/nnano.2007.260","article-title":"Intrinsic peroxidase-like activity of ferromagnetic nanoparticles","volume":"2","author":"Gao","year":"2007","journal-title":"Nat. Nanotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"6060","DOI":"10.1039\/c3cs35486e","article-title":"Nanomaterials with enzyme-like characteristics (nanozymes): Next-generation artificial enzymes","volume":"42","author":"Wei","year":"2013","journal-title":"Chem. Soc. Rev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"123546","DOI":"10.1016\/j.talanta.2022.123546","article-title":"Enzyme-controllable just-in-time production system of copper hexacyanoferrate nanoparticles with oxidase-mimicking activity for highly sensitive colorimetric immunoassay","volume":"247","author":"Lai","year":"2022","journal-title":"Talanta"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11302","DOI":"10.1021\/jacs.8b05223","article-title":"Catalytically synthesized prussian blue nanoparticles defeating natural enzyme peroxidase","volume":"140","author":"Komkova","year":"2018","journal-title":"J. Am. Chem. Soc."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"106489","DOI":"10.1016\/j.microc.2021.106489","article-title":"Enhanced peroxidase-like activity of copper phosphate modified by hydrophilic phytic-acid and its application in colorimetric detection of hydrogen peroxide","volume":"168","author":"Zhang","year":"2021","journal-title":"Microchem. J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"129744","DOI":"10.1016\/j.colsurfa.2022.129744","article-title":"The peroxidase-like catalytic activity of in situ prepared cobalt carbonate and its applications in colorimetric detection of hydrogen peroxide, glucose and ascorbic acid","volume":"651","author":"Peng","year":"2022","journal-title":"Colloids Surf. A Physicochem. Eng. Asp."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"108669","DOI":"10.1016\/j.bej.2022.108669","article-title":"Enzyme-integrated biomimetic cobalt metal-organic framework nanozyme for one-step cascade glucose biosensing via tandem catalysis","volume":"188","author":"Goud","year":"2022","journal-title":"Biochem. Eng. J."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"131720","DOI":"10.1016\/j.snb.2022.131720","article-title":"A novel colorimetric strategy for rapid detection of dimethoate residue in vegetables based on enhancing oxidase-mimicking catalytic activity of cube-shape Ag2O particles","volume":"361","author":"Zhan","year":"2020","journal-title":"Sens. Actuator B-Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.aca.2020.12.019","article-title":"Ratiometric fluorescence sensing of glutathione by using the oxidase-mimicking activity of MnO2 nanosheet","volume":"1145","author":"Cao","year":"2021","journal-title":"Anal. Chim. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3683","DOI":"10.1039\/C8CS00718G","article-title":"Nanozyme: New horizons for responsive biomedical applications","volume":"48","author":"Jiang","year":"2019","journal-title":"Chem. Soc. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6627","DOI":"10.1007\/s00216-021-03629-5","article-title":"A sensitive photothermometric biosensor based on redox reaction-controlled nanoprobe conversion from Prussian blue to Prussian white","volume":"413","author":"Zhang","year":"2021","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"121564","DOI":"10.1016\/j.saa.2022.121564","article-title":"Modulation of inner filter effect between persistent luminescent particles and 2, 3-diaminophenazine for ratiometric fluorescent assay of ascorbic acid and ascorbate oxidase activity","volume":"280","author":"Yao","year":"2022","journal-title":"Spectrochim. Acta. A Mol. Biomol. Spectrosc."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1007\/s00604-019-3580-9","article-title":"Manganese(II)-doped zinc\/germanium oxide nanoparticles as a viable fluorescent probe for visual and time-resolved fluorometric determination of ascorbic acid and its oxidase","volume":"186","author":"Han","year":"2019","journal-title":"Microchim. Acta"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1039\/C9AN02108F","article-title":"A fluorescence \u201coff\u2013on\u2013off\u201d sensing platform based on bimetallic gold\/silver nanoclusters for ascorbate oxidase activity monitoring","volume":"145","author":"Wang","year":"2020","journal-title":"Analyst"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1156","DOI":"10.1039\/D2NJ05460D","article-title":"Investigating the oxidase-like activity of a Co\u2013Fe Prussian blue analogue nanocube prepared in situ and its applications in the colorimetric detection of ascorbic acid, alkaline phosphatase, \u03b1-glucosidase, and ascorbic acid oxidase","volume":"47","author":"Peng","year":"2023","journal-title":"New J. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.aca.2020.01.035","article-title":"Colorimetric acid phosphatase sensor based on MoO3 nanozyme","volume":"1105","author":"Lin","year":"2020","journal-title":"Anal. Chim. Acta"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"339717","DOI":"10.1016\/j.aca.2022.339717","article-title":"Constructing bifunctional metaleorganic framework based nanozymes with fluorescence and oxidase activity for the dualchannel detection of butyrylcholinesterase","volume":"1205","author":"Wang","year":"2022","journal-title":"Anal. Chim. Acta"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"9318","DOI":"10.1021\/acsanm.0c01965","article-title":"Gold-platinum bimetallic nanoclusters for oxidase-like catalysis","volume":"3","author":"Zhang","year":"2020","journal-title":"ACS Appl. Nano Mater."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5827","DOI":"10.1021\/jf500950p","article-title":"Colorimetric detection of sulfite in foods by a TMB-O2-Co3O4 nanoparticles detection system","volume":"62","author":"Qin","year":"2014","journal-title":"J. Agric. Food Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5076","DOI":"10.1021\/acssuschemeng.9b07009","article-title":"Blue light-gated reversible silver nanozyme reaction networks that achieve life-like adaptivity","volume":"8","author":"Jiang","year":"2020","journal-title":"ACS Sustain. Chem. Eng."}],"container-title":["Biosensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6374\/13\/3\/344\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:47:49Z","timestamp":1760122069000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6374\/13\/3\/344"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,4]]},"references-count":33,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["bios13030344"],"URL":"https:\/\/doi.org\/10.3390\/bios13030344","relation":{},"ISSN":["2079-6374"],"issn-type":[{"value":"2079-6374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,4]]}}}