The application of new UPLC-MS/MS multimethod of antimicrobials residues determination for the control of honey safety
The article presents the results of the developed UPLC-MS/MS multimethod application for quantitative determination of different groups of antimicrobials residues in honey. According to the European RASFF system, despite the ban on the use of antimicrobials in beekeeping, there are still many reports of antibiotics detection in honey. Today there are no harmonized global requirements for honey safety, which are to be mandatory for honey exports and imports worldwide, and thus the instrumental abilities of the control laboratory (e.g., CCα and CCβ) often are the main criteria of honey batches compliance. The development and use of multianalyte methods have obvious advantages, viz. shortened time of the analysis, reduced consumption of necessary consumables and reagents, the decrease of utilization time of laboratory equipment etc. Main advantages of our developed multimethod are the simplicity, rapidity and cost-effectiveness, since the analysis is performed in one sample preparation without the use of filtration or solid-phase extraction. This method provides selective determination of target analytes with LODs in the ranges from 0.05 μg/kg for chloramphenicol to 2 μg/kg for sulfanilamide and tetracycline; and it exceeds the requirements of the MRPLs or regulatory limits provided with a sufficient level of precision of the method and a high percentage of recoveries, which is in accordance with the requirements of Decision 2002/657/EC. The technique was tested in the analysis of natively contaminated and fortified honey samples, as well as in the interlaboratory studies.
During the period of 2018-2019, 809 export lots of honey were analyzed for safety, as well as 946 samples were tested during honey incoming control using our developed multimethod. The most frequently investigated analytes in honey were metronidazole and chloramphenicol, viz. 68% and 52% of the analyzed samples, respectively. This may be related to the risks of unauthorized use by apiaries of human medicine drugs as well as the counterfeit veterinary preparations containing these antibiotics, but which are not declared. With regard to chloramphenicol, the main reason for detecting its residues in honey today is the illegal use of preparations containing a mixture of chloramphenicol and its SS-para-stereoisomer dextramycin. The isomer has no therapeutic effect, but has repeatedly been found in counterfeit preparations for bees and, as a consequence, in honey.
It was found that among the samples for incoming honey control 297 contained target analytes at the level above the detection limit of the method. Most often, viz. in 28% of cases, the regulatory limit (3 μg/kg) was exceeded for sulfathiazole, which may be used to treat and prevent American foulbrood, one of the most dangerous bee diseases. In few cases, the analyzed homogenized honey samples contained simultaneously five or six target analytes at concentration levels above their LOD. This clearly demonstrates the fact that Ukrainian beekeepers occasionally use unregistered antimicrobials for the treatment and/or prevention of bee diseases. The data obtained do not claim to be highly representative, since they are based on the analysis of honey samples directly from the customers and not obtained as a result of monitoring by random sampling.
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