Development of an HPLC-MS/MS method for the quantitative determination of chloramphenicol in honey from Kazakhstan

Addressing the issue of antibiotics, particularly chloramphenicol, in bee products is an urgent concern within the trade of these items due to potential risks to human health. Developing highly sensitive techniques for detecting trace amounts of antibiotic residues is crucial. This study aimed to establish an analytical method using HPLC-MS/MS (High-Performance Liquid Chromatography – Tandem Mass Spectrometry) for the identification and quantification of residual chloramphenicol in honey samples sourced from Kazakhstan. The developed method involved ultrasonic extraction followed by HPLC-MS/MS with electrospray ionisation in negative mode. Multiple reaction monitoring was utilised in configuring the parameters of the tandem mass spectrometric detector. The proposed procedure offered notable advantages, such as the simplicity and speed of the extraction process, as well as the sensitivity and accuracy of the detection method. The limit of detection achieved is 0.09 µg/kg, with a linear calibration curve spanning the concentration range of 0.1-1.0 µg/kg and a correlation coefficient of 0.9985. The extraction rate ranged between 97% and 101%. Importantly, this HPLC-MS/MS method for chloramphenicol determination in honey surpassed the maximum residue limit requirements (0.3 µg/kg) in terms of sensitivity, while maintaining methodological precision and a high extraction percentage. This compliance aligned with the standards outlined in Commission Decision 2002/657/EC. The method was applied to 14 honey samples from Kazakhstan, all of which were found to have chloramphenicol levels below the method’s detection limit. However, when testing imported honey samples, two out of seven exceeded the detection limit for chloramphenicol. The developed HPLC-MS/MS method for detecting chloramphenicol residues in honey samples can serve as a valuable tool for routine laboratory analysis and for scrutinising potentially suspicious or questionable samples