4.7 Article

Competitive immunoassay using enzyme-regulated Fe3O4@COF/Fe3+ fluorescence probe for natural chloramphenicol detection

Journal

ANALYTICA CHIMICA ACTA
Volume 1277, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.aca.2023.341680

Keywords

Magnetic covalent organic frameworks; Fluorescence sensor; Enzyme-linked immunoassay; Competitive assay; Chloramphenicol

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Accurate and sensitive detection of chloramphenicol (CAP) in natural samples is achieved using an enzyme-regulated fluorescence sensor. A Fe3O4@COF/Fe3+ probe is developed, where Fe3O4@COF acts as a magnetic carrier and signal probe. The sensor demonstrates good stability, reusability, and a logarithmic linear relationship between CAP concentration and fluorescence intensity, providing a reliable method for CAP detection with a low limit of detection.
Accurate and sensitive detection of chloramphenicol (CAP) in natural samples is essential for ensuring human health. Herein, an enzyme-regulated fluorescence sensor using Fe3O4@COF/Fe3+ probe, is developed for CAP determination. Fe3O4@COF, synthesized via hydrothermal method, exhibits dual functions as a magnetic carrier and signal probe. Bovine serum albumin conjugated-chloramphenicol, adsorbed on the surface of Fe3O4@COF, competes with CAP for antibody binding. The antibody interacts with alkaline phosphatase via the biotinstreptavidin system. Meanwhile, ascorbic acid, produced from the enzyme-catalyzed reaction dominated by alkaline phosphatase, effectively restores the fluorescence of Fe3O4@COF that is quenched by Fe3+. After experimental verification and gradual optimization, a logarithmic linear relationship between CAP concentration and fluorescence intensity is established in the range of 2 x 10(-4) -10 mu g mL(-1), with a good limit of detection (9.2 x 10(-5) mu g mL(-1)). Proposed method exhibits excellent stability (15 days) and reusability (8 cycles), providing a sensitive and reliable method for accurate CAP detection. The readouts show good agreement with HPLC and recoveries during laboratory and natural CAP analysis.

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