Monitoring of florfenicol residues in fish muscle by HPLC-UV with confirmation of suspect results by LC-MS/MS

• Published in: Drug testing and analysis Vol. 4; no. S1; pp. 125 - 129
• Main Authors: Granja, Rodrigo H. M. M., de Lima, Andreia C., Patel, Raj K., Salerno, Alessandro G., Wanschel, Amarylis C. B. A.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-08-2012
• Subjects: Animals; Anti-Bacterial Agents - analysis; Anti-Bacterial Agents - isolation & purification; Brazil; Chromatography, Liquid - methods; Drug Residues - analysis; Drug Residues - isolation & purification; fish; Fishes - metabolism; florfenicol; high performance liquid chromatography; liquid chromatography-tandem mass spectrometry; Muscles - metabolism; Reproducibility of Results; residues; Seafood - analysis; Solid Phase Extraction - methods; Tandem Mass Spectrometry - methods; Thiamphenicol - analogs & derivatives; Thiamphenicol - analysis; Thiamphenicol - isolation & purification; Brazil
• ISSN: 1942-7603; 1942-7611
• DOI: 10.1002/dta.1362

Florfenicol, a derivative of thiamphenicol in which the hydroxyl group at C‐3 has been replaced with fluorine, is listed by the World Health Organization as an antibacterial agent for human medicine that is critically important in risk management strategies for non‐human use. AOAC International has also identified it as an important molecule for the development of effective methods for the seafood sector. Following inspection missions from the European Union and United States of America, it was introduced in the Brazilian Residues Control Program to fulfill export and internal national requirements with a Maximum Residue Limit of 800 µg/kg. A high performance liquid chromatography method with ultraviolet detection at a wavelength of 230 nm (λ = 230 nm) for the detection of florfenicol in fish muscle was developed and validated according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC). Fish samples were extracted with ethyl acetate and hexane followed by C18 solid phase clean‐up and chromatographic separation on a reversed‐phase C18 LC column with acetonitrile:water as mobile phase. The method results were also was compared with those obtained using liquid chromatography‐tandem quadrupole mass spectrometry. The method meets the Brazilian regulatory requirements with a decision limit (CCα) of 840 µg/kg and detection capability (CCβ) of 879 µg/kg. This method is easy to use and has been implemented into Brazil's residue control program, with liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) confirmation of any suspect samples using the same method. © 2012 John Wiley & Sons, Ltd.
Florfenicol is a broad‐spectrum antibiotic and widely used in veterinary products. Residues of florfenicol in foods of animal origin, including those raised in aquaculture intended for human consumption is of toxicological and regulatory concern. The aim of this study was to develop a simple, rapid, and reliable high performance liquid chromatography method with ultraviolet detection (HPLC/UV) for the determination of florfenicol in fish muscle. The eventual identity of florfenicol found in routine samples was confirmed by LC‐MS/MS.