A new colorimetric lactate biosensor based on CUPRAC reagent using binary enzyme (lactate-pyruvate oxidases)-immobilized silanized magnetite nanoparticles

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dc.authoridDilgin, Yusuf/0000-0002-2980-6871
dc.authoridAPAK, Resat/0000-0003-1739-5814
dc.contributor.authorAyaz, Selen
dc.contributor.authorErsan, Teslime
dc.contributor.authorDilgin, Yusuf
dc.contributor.authorApak, Resat
dc.date.accessioned2025-01-27T20:52:02Z
dc.date.available2025-01-27T20:52:02Z
dc.date.issued2024
dc.departmentÇanakkale Onsekiz Mart Üniversitesi
dc.description.abstractA novel optical lactate biosensor is presented that utilizes a colorimetric interaction between H2O2 liberated by a binary enzymatic reaction and bis(neocuproine)copper(II) complex ([Cu(Nc)2]2+) known as CUPRAC (cupric reducing antioxidant capacity) reagent. In the first step, lactate oxidase (LOx) and pyruvate oxidase (POx) were separately immobilized on silanized magnetite nanoparticles (SiO2@Fe3O4 NPs), and thus, 2 mol of H2O2 was released per 1 mol of the substrate due to a sequential enzymatic reaction of the mixture of LOx-SiO2@Fe3O4 and POx-SiO2@Fe3O4 NPs with lactate and pyruvate, respectively. In the second step, the absorbance at 450 nm of the yellow-orange [Cu(Nc)2]+ complex formed through the color reaction of enzymatically produced H2O2 with [Cu(Nc)2]2+ was recorded. The results indicate that the developed colorimetric binary enzymatic biosensor exhibits a broad linear range of response between 0.5 and 50.0 mu M for lactate under optimal conditions with a detection limit of 0.17 mu M. The fabricated biosensor did not respond to other saccharides, while the positive interferences of certain reducing compounds such as dopamine, ascorbic acid, and uric acid were minimized through their oxidative removal with a pre-oxidant (NaBiO3) before enzymatic and colorimetric reactions. The fabricated optical biosensor was applied to various samples such as artificial blood, artificial/real sweat, and cow milk. The high recovery values (close to 100%) achieved for lactate-spiked samples indicate an acceptable accuracy of this colorimetric biosensor in the determination of lactate in real samples. Due to the increase in H2O2 production with the bienzymatic lactate sensor, the proposed method displays double-fold sensitivity relative to monoenzymatic biosensors and involves a neat color reaction with cupric-neocuproine having a clear stoichiometry as opposed to the rather indefinite stoichiometry of analogous redox dye methods.
dc.description.sponsorshipTrkiye Bilimsel ve Teknolojik Arascedil;timath;rma Kurumu
dc.description.sponsorshipNo Statement Available
dc.identifier.doi10.1007/s00604-024-06531-w
dc.identifier.issn0026-3672
dc.identifier.issn1436-5073
dc.identifier.issue8
dc.identifier.pmid38980437
dc.identifier.scopus2-s2.0-85197820390
dc.identifier.scopusqualityQ1
dc.identifier.urihttps://doi.org/10.1007/s00604-024-06531-w
dc.identifier.urihttps://hdl.handle.net/20.500.12428/25633
dc.identifier.volume191
dc.identifier.wosWOS:001265799100002
dc.identifier.wosqualityN/A
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherSpringer Wien
dc.relation.ispartofMicrochimica Acta
dc.relation.publicationcategoryinfo:eu-repo/semantics/openAccess
dc.rightsinfo:eu-repo/semantics/openAccess
dc.snmzKA_WoS_20250125
dc.subjectColorimetric biosensor
dc.subjectCUPRAC reagent
dc.subjectBienyzmatic biosensor
dc.subjectMagnetite nanoparticle
dc.subjectLactate biosensors
dc.titleA new colorimetric lactate biosensor based on CUPRAC reagent using binary enzyme (lactate-pyruvate oxidases)-immobilized silanized magnetite nanoparticles
dc.typeArticle

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