Determination of tetracycline residues in honey

Abstract

Antibiotics from the tetracycline group are widely used for prevention and control of infectious diseases and have a great activity against variety of Gram-positive and Gram-negative bacteria. Due to the widespread use of tetracyclins in animal husbandry, it can lead to an increase the risk of antibiotics remaining in human food. In this study, the degree of contamination of honey from the markets with antibiotics from the tetracycline group was examined. Antibiotics from the tetracycline group: Tetracyclin, Oxitetracyclin and Chlortetracyclin were examined by competitive enzyme immunoassay method for the quantitative analysis using the kits R-biopharm, Biopanda Reagen oxitetracyclin and chlortetracyclin. The principle of the method is to identify the unknown amount of antibiotic by present in the sample and the fixed amount of antibiotic antigens pre-coated on the wells of microtiter strips compete for the antiantibiotics antibodies, which in turn is detected with enzyme conjugate. After incubation the wells are washed and the bound enzyme is visualised by adding TMB solution. Any coloured product is measured at 450 nm after adding stop solution. The absorbance value of the developed colour is inversely proportional to the amount of the antibiotics in the sample. The quantity of antibiotics in the test sample can be interpolated from the standard curve constructed from the standards, and corrected for sample dilution.