Abstract

An amperometric magneto-immunosensor (AMIS) for the detection of residues of fluoroquinolone antibiotics in milk samples is described for the first time. The immunosensor presented combines magnetic beads biomodified with an antibody with a broad recognition profile of fluoroquinolones, a haptenized enzyme and a magnetic graphite–epoxy composite (m-GEC) electrode. After the immunochemical reaction with specific enzyme tracer, the antibody biomodified magnetic beads are easily captured by an electrode made of graphite-epoxy composite containing a magnet, which also acts as transducer for the electrochemical detection. In spite of the complexity of milk, the use of magnetic beads allows elimination of potential interferences caused by the matrix components; hence the AMIS could perform quantitative measurements, directly in these samples, without any additional sample cleanup or extraction step. The immunosensor is able to detect up to seven different fluoroquinolones far below the MRLs defined by the UE for milk; for example ciprofloxacin is detected directly in milk with an IC50 of 0.74 μg/L and a LOD of 0.009 μg/L. This strategy offers great promise for rapid, simple, cost-effective, and on-site analysis fluoroquinolones in complex samples.

Highlights

  • In modern intensive farming systems, the introduction of veterinary pharmaceuticals, pesticides and complex animal feeds has been critical to enhance the productivity by increasing the density of animals in the farms

  • Because of the concerns regarding drug residues entering the food chain and contaminating environment, and contributing to bacterial resistance development, maximum residue levels (MRLs) for the detection of FQ in several matrices from diverse animal species have been established by the European Commission (Council regulation 2377/90/EC [5] and council directive 96/23/EC [6]), as well as the requirements of the analytical methods that public health control and veterinary laboratories should use for this purpose (Commission decision 2002/657/EC [7])

  • The AIMS presented here, is based on the use of an enzyme tracer, magnetic beads modified with antifluoroquinolone antibodies, and graphite–epoxy composite electrodes containing a magnet inside (m-GEC electrodes)

Read more

Summary

Introduction

In modern intensive farming systems, the introduction of veterinary pharmaceuticals, pesticides and complex animal feeds has been critical to enhance the productivity by increasing the density of animals in the farms. Because of the concerns regarding drug residues entering the food chain and contaminating environment, and contributing to bacterial resistance development, maximum residue levels (MRLs) for the detection of FQ in several matrices from diverse animal species have been established by the European Commission (Council regulation 2377/90/EC [5] and council directive 96/23/EC [6]), as well as the requirements of the analytical methods that public health control and veterinary laboratories should use for this purpose (Commission decision 2002/657/EC [7]). Analytical methods, with accurate and fast responses without sample pre-treatment, opening up the possibility of direct on-site analysis with intuitive devices [13] In this sense, electrochemical immunosensors combine the high sensitivity of the electrochemical transducers with the high selectivity of antibodies, becoming powerful analytical tools for the detection of antibiotic residues [14,15]. Fluoroquinolone residues can be directly detected in milk samples, without the need of previous cleanup or purification steps

Materials and Instruments
Chemicals and Immunochemicals
Buffers and Solutions
Preparation of the Enzyme Labeled Immunoreagents
Preparation of the Biomodified Magnetic Beads
Specificity Studies
2.10. Milk Matrix Effect
Results and Discussion
Preparation and Characterization of Enzyme Labeled Immunoreagents
Preparation and Characterization of Biomodified Magnetic Beads
Specificity Assay
Milk Matrix Effect
Amperometric Magneto Immunosensor
Conclusions
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call