Development of a carbon paper-based electrochemical immunosensor for trimethoprim detection in fishery species: monitoring antibiotic contaminants: Original scientific article

Maria Freitas; Vitória  Dibo; Rita Ribeiro; Cristina Delerue-Matos; Simone Morais; Álvaro Torrinha

doi:10.5599/admet.2962

Authors

Maria Freitas REQUIMTE/LAQV, ISEP, Polytechnic of Porto, Rua Dr. António Bernardino de Almeida, 431, 4249-015, Porto, Portugal https://orcid.org/0000-0002-0628-2753
Vitória Dibo REQUIMTE/LAQV, ISEP, Polytechnic of Porto, Rua Dr. António Bernardino de Almeida, 431, 4249-015, Porto, Portugal https://orcid.org/0000-0003-0354-2860
Rita Ribeiro REQUIMTE/LAQV, ISEP, Polytechnic of Porto, Rua Dr. António Bernardino de Almeida, 431, 4249-015, Porto, Portugal https://orcid.org/0009-0008-3085-7079
Cristina Delerue-Matos REQUIMTE/LAQV, ISEP, Polytechnic of Porto, Rua Dr. António Bernardino de Almeida, 431, 4249-015, Porto, Portugal https://orcid.org/0000-0002-3924-776X
Simone Morais REQUIMTE/LAQV, ISEP, Polytechnic of Porto, Rua Dr. António Bernardino de Almeida, 431, 4249-015, Porto, Portugal https://orcid.org/0000-0001-6433-5801
Álvaro Torrinha REQUIMTE/LAQV, ISEP, Polytechnic of Porto, Rua Dr. António Bernardino de Almeida, 431, 4249-015, Porto, Portugal https://orcid.org/0000-0002-9829-8559

DOI:

https://doi.org/10.5599/admet.2962

Keywords:

Amperometry, biosensor, paper-based electrode, pharmaceuticals, drug analysis, aquatic ecosystem

Abstract

Background and purpose: Monitoring antibiotic drugs in the environment is particularly relevant given their role in fostering microbial resistance, impacting aquatic species and human health. Therefore, this work addresses the lack of a sustainable and cost-effective analytical approach and reports the development of a competitive electrochemical immunosensor for the rapid analysis of trimethoprim (TMP), an aquatic contaminant of emerging concern, overcoming the limitations of conventional methods that are often costly and time-consuming. Experimental approach: A miniaturized 3-electrode system was developed to support the analyses of a low-volume sample (200 µL), using a low-cost carbon-paper working electrode, pencil graphite lead auxiliary electrode and a stainless-steel needle reference electrode. Scanning electron microscopy was used to characterize the transducer, revealing a network of randomly arranged carbon fibres, significant for efficient antibody immobilization. For the immunoassay construction, an anti-TMP antibody was physically adsorbed to a small-sized transducer (d = 4 mm) for the specific recognition of TMP, followed by incubation with the enzyme-conjugate (horseradish peroxidase) and the enzyme-substrate (tetramethylbenzidine). Key results: The analytical signal was recorded by chronoamperometry (1-minute reaction) and yielded a limit of detection of 34 ng L⁻¹. The 45-minute assay demonstrates accuracy (92.0 to 103.2 % in fishery species including codfish, mackerel, crab, and hake), reproducibility (6.1 and 9.3 % for repeatability and inter-day variation coefficient) and high selectivity (bias less than 5 %) analysis. The sensor's performance was validated against a conventional Enzyme-Linked Immunosorbent Assay. Conclusion: This study introduces a sustainable electrochemical immunosensor, offering a portable and eco-friendly alternative for the rapid detection of TMP in fishery species, addressing the limitations of traditional methods.

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