Detection of tetracycline antibiotics in honey using high-performance liquid chromatography

Abstract

It is hard to overestimate the value of honey since the constituent substances in it are of great importance in the food industry and medicine. The quality of honey is established by the legislation of the Russian Federation (GOST). But the levels of these standards are regulated by outdated methods of analysis, which can not give reliable results. The detection of antibiotic residues in honey is a central issue in the quality and safety control of this product. Accumulation of drugs in honey used to treat bee colonies can cause allergies and dysbiosis in people who have eaten such honey, as well as develop antibiotic resistance in microorganisms. At present, one of the promising directions in the field of detecting medicines in honey is the use of high-performance liquid chromatography (HPLC). It helps selectively and accurately detect antibiotic substances in honey bee products. In Russia, the HPLC method is used with mass spectrometry to detect tetracycline residues in honey. A significant drawback with this method that limits the widespread use of it is the application of expensive, technically sophisticated equipment that needs high-quality reagents and consumables. The purpose of this work was to develop the simultaneous identification of tetracycline antibiotics in honey and carry out their quantitative analysis by a reversed-phase HPLC method. A sample preparation algorithm was developed, and the conditions for chromatographing combined indication of tetracyclines in honey at an acceptable concentration according to the MRL (0.01 mg/kg) with Agilent 1260 Infiniti liquid chromatograph equipped with a column thermostat, a gradient pump, and a UV detector were selected. According to the results of the research, the most optimal condition for the simultaneous analysis of oxytetracycline, tetracycline hydrochloride, chlortetracycline in honey by HPLC method with ultraviolet detection was a wavelength of 254 nm, a flow rate of 0.5 ml/min and a column temperature of 25° C. Under these conditions, the antibiotic retention time was determined: 4.069 minutes for oxytetracycline, 4.331 minutes for tetracycline hydrochloride, 4.642 minutes for chlortetracycline. The developed HPLC method for the simultaneous determination of tetracycline antibiotics in honey was tested on honey bee products from the regions of the Republics of Tatarstan and Bashkortostan.