Solid-phase clean-up and thin-layer chromatographic detection of veterinary aminoglycosides

Area of Science:

• Veterinary Pharmacology
• Analytical Chemistry
• Food Safety

Background:

• Microbial inhibition assays can detect antibiotics in farm animals, but chemical confirmation is essential.
• Accurate quantification of antibiotic residues in animal tissues and fluids is crucial for food safety and animal health.
• Aminoglycoside antibiotics are widely used in veterinary medicine, necessitating reliable detection methods.

Purpose of the Study:

• To develop and optimize a chemical method for isolating and detecting hygromycin B in animal biological fluids.
• To establish the sensitivity and applicability of the method for semi-quantitative estimation of aminoglycosides.
• To enable the confirmation of antibiotic residues detected by initial screening assays.

Main Methods:

• Optimized solid-phase extraction using copolymeric bonded silica columns for hygromycin B isolation.
• Thin-layer chromatography (TLC) for separation of antibiotic compounds.
• Fluorescence detection of hygromycin B after derivatization with fluorescamine.
• Analysis of bovine plasma and swine serum samples fortified with hygromycin B.

Main Results:

• The optimized method achieved a detection limit of 50 ng for hygromycin B.
• Hygromycin B was successfully detected in bovine plasma and swine serum at various concentrations (25-500 ng/ml).
• The method demonstrated the ability to separate and detect multiple aminoglycosides, including gentamicin, neomycin, spectinomycin, hygromycin B, and streptomycin, by TLC.

Conclusions:

• The developed procedure is a rapid and sensitive method for the semi-quantitative estimation of aminoglycosides in animal samples.
• This chemical method provides reliable confirmation for antibiotic residues, complementing microbial inhibition assays.
• The technique is valuable for ensuring food safety and monitoring antibiotic usage in livestock.