Immobilized acetylcholinesterase in magnetic nanoparticles for in-line inhibition studies using a capillary electrophoresis system.

Autor: Siebert DA; Laboratório de Eletroforese Capilar, Departamento de Química, Universidade Federal de Santa Catarina, Trindade, CEP 88040-900, Florianópolis, SC, Brazil., Caon NB; Laboratório de Estudo em Materiais Poliméricos, Departamento de Química, Universidade Federal de Santa Catarina, Trindade, CEP 88040-900, Florianópolis, SC, Brazil., Alberton MD; Laboratório de Pesquisa em Produtos Naturais, Universidade Regional de Blumenau, Rua São Paulo 2171, CEP 89030-000, Blumenau, SC, Brazil., Vitali L; Laboratório de Eletroforese Capilar, Departamento de Química, Universidade Federal de Santa Catarina, Trindade, CEP 88040-900, Florianópolis, SC, Brazil., Parize AL; Laboratório de Estudo em Materiais Poliméricos, Departamento de Química, Universidade Federal de Santa Catarina, Trindade, CEP 88040-900, Florianópolis, SC, Brazil., Micke GA; Laboratório de Eletroforese Capilar, Departamento de Química, Universidade Federal de Santa Catarina, Trindade, CEP 88040-900, Florianópolis, SC, Brazil. Electronic address: gustavo.micke@ufsc.br.
Jazyk: angličtina
Zdroj: Analytica chimica acta [Anal Chim Acta] 2023 Sep 22; Vol. 1275, pp. 341566. Date of Electronic Publication: 2023 Jun 27.
DOI: 10.1016/j.aca.2023.341566
Abstrakt: Enzyme assays can be performed with the capillary electrophoresis technique (CE) in many approaches, such as the immobilized enzyme micro-reactor. Acetylcholinesterase is a promising enzyme to be used when pursuing such a method, as it has already been explored in the proposal of similar methods of miniaturizing enzyme assays. The present work proposes a novel enzyme micro-reactor, based on the anchorage of the enzyme on magnetic nanoparticles of MnFe 2 O 4, with chitosan and glutaraldehyde as the cross-linker in the capillary by means of an arrange of neodymium magnets. The calculated K m of the enzyme evaluated by this method was 1.12 mmol L -1 , comparable to other studies in the literature that utilizes immobilized enzymes. Also, IC 50 for neostigmine was assessed in 3 different micro-reactors, with an average of 29.42 ± 3.88 μmol L -1 . In terms of the micro-reactor stability, it was possible to perform at least 25 experiments with assembled micro-reactor. The method was applied to hydroalcoholic extracts of 7 plant species. Plinia cauliflora had the best result, with 42.31 ± 6.81% of enzyme inhibition in a concentration of 100 mg L -1 .
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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