Validation of a rapid and sensitive routine method for determination of chloramphenicol in honey by LC–MS/MS

Abstract

Chloramphenicol (CAP) is a broad spectrum antibiotic used in the treatment of human and animal diseases. However, CAP can exhibit toxic effects in certain susceptible individuals, causing bone marrow depression, including fatal aplastic anemia. As this condition is dose-independent, CAP has been banned for use in food-producing animals, including honeybees. In this study, a quick, simple and low-cost routine analytical method was developed for the screening and confirmation of chloramphenicol in honey by LC–MS/MS. Sample clean-up takes only two steps without SPE procedure and with recoveries >97%. Honey samples were selected from several producers in Brazil and diluted in a small amount of water. After fortification and addition of d _s-chloramphenicol as internal standard, the samples were extracted with ethyl acetate. Complete validation of the method was performed on the basis of EU decision 2002/657. Within-laboratory CV reproducibility at the lowest concentration was <10%. An evaluation of two different methods to calculate the decision limit and detection capability gave 0.08 µg kg⁻¹ for CCα and 0.12 µg kg⁻¹ for CCβ.

Keywords:

• honey
• veterinary drug residues
• chloramphenicol
• chromatography
• LC/MS

Acknowledgements

The authors thank Aparecida Rocha Chagas for technical assistance, Applied Biosystems for the technical support and Brazilian Ministry of Agriculture for financial support.