Abstract
Background: Antibiotic residues are a problem of increasing importance and have direct consequences for human and animal health. The frequent use of antibiotics in veterinary practice causes their excretion in milk in dairy cattle. This way, they can easily enter the human body through the consumption of milk and dairy products. Objectives: This induces the need for accurate and sensitive methods to monitor antibiotic levels in milk. The aim of this study was to develop a rapid and sensitive magnetic nanoparticle-based fluorescence immunoassay for the simultaneous detection of chloramphenicol and penicillin G in milk. Methods: Magnetic nanoparticles were synthesized and functionalized with (3-aminopropyl)triethoxysilane. Chloramphenicol-Ovalbumin and Chloramphenicol-Ovalbumin-Fluorescein-5-isothiocyanate conjugates were prepared. Penicillin G – ATTO 633 fluorescent conjugate was synthesized. Antibodies against chloramphenicol and penicillin G were immobilized onto the magnetic nanoparticles. The competitive fluorescent immunoassay was developed. The optimal concentration of the antibody-magnetic nanoparticles and the fluorescent conjugates for the assay was determined. The calibration curves for the antibiotics in buffer and milk were plotted. Fluorescent immunoassay for the simultaneous determination of chloramphenicol and penicillin G in milk was developed. Results: The limit of detection by the simultaneous immunoassay of chloramphenicol and penicillin G in milk was 0.85 ng/mL and 1.6 ng/mL, respectively. The recovery of different concentrations of chloramphenicol and penicillin G in milk samples varied from 98% to 106%. Conclusions: A rapid and sensitive magnetic nanoparticle-based immunofluorescent assay for the simultaneous determination of chloramphenicol and penicillin G in milk was developed. The magnetic nanoparticles ensured rapid and easy procedure.
Highlights
Antibiotics are widely used for disease prevention and treatment as well as growth promotion in animal husbandry [1]
The recovery of different concentrations of chloramphenicol and penicillin G in milk samples varied from 98% to 106%
A rapid and sensitive magnetic nanoparticle-based immunofluorescent assay for the simultaneous determination of chloramphenicol and penicillin G in milk was developed
Summary
Antibiotics are widely used for disease prevention and treatment as well as growth promotion in animal husbandry [1]. The most used conventional methods for the detection of antibiotics are instrumental [7], such as capillary electrophoresis [8, 9], gas chromatography [10, 11], and liquid chromatography [12] coupled with mass spectrometry [13] These methods are expensive and present limitations, such as the need for expensive laboratory instruments, time-consuming separation/clean-up methodologies, long analysis time, extensive sample handling with multiple washing steps, use of expensive and polluting solvents and the impossibility to perform field analyses, making necessary the development of new methods to overcome these limitations [14]. Immunoassays based on specific antibody recognition are a rapid method of screening samples [15] They are simple, fast selective, sensitive, and low-cost assays. This way, they can enter the human body through the consumption of milk and dairy products
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