Extraction Strategies for Simultaneous Determination of Florfenicol and Florfenicol Amine in Tilapia (Oreochromis niloticus) Muscle: Quantification by LC-MS/MS
A liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of the veterinary drug florfenicol (FF) and its major metabolite, florfenicol amine (FFA), in tilapia muscles (Oreochromis niloticus). Three different sample preparation procedures (DLLME, sub-zero, and modified QuEChERS) were tested. The best extraction results were obtained by using the modified QuEChERS. The quantification was made by using a LC-MS/MS analysis, with a Lichrocart Cartridge Purospher Star C8 HPLC column (250 mm×4.6 mm, 5 μm particle size). Analytes were separated with a mobile phase consisting of Milli-Q water to acetonitrile 40:60 (v/v), both with 0.1% formic acid. The validation parameters were recovery of 70 to 79% and 62 to 69%, limit of detection of 0.0625 μg g−1 and 0.125 μg g−1, and limit of quantification of 0.125 μg g−1 and 0.25 μg g−1, for FF and FFA, respectively. CCα was 1183 μg kg−1 and CCβ was 1365 μg kg−1 for FF, intraday and interday precision has CV ≤20%, and linear range was 0.625 to 5.00 μg g−1. This method was shown to be simple and rapid when compared to other, more conventional methods. Also, it has low reagent and solvent consumption, with low waste generation, which is in line with the principles of green chemistry. The method was successfully applied for the analyzes of tilapia exposed to the antibiotic.
KeywordsVeterinary drugs Green chemistry Residues Antibiotics Aquaculture
Limit of decision
Ministry of Agriculture, Livestock and Food Supply - Brazil
Molecularly imprinted solid-phase extraction
Matrix solid-phase dispersion extraction
Quick, Easy, Cheap, Effective, Rugged and Safe
Ultrasound-assisted dispersive liquid-liquid microextraction
The authors wish to thank Prof. Carol Collins for language assistance.
This research was supported by BNDES (0117020010606007), FAPESP (2014/50867-3), CNPq (311671/2015-2; 465389/2014-7) and INCT Bioanalítica. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil, Finance Code 001.
Compliance with Ethical Standards
Conflict of Interest
Letícia Sayuri Shiroma, Sonia Claudia do Nascimento de Queiroz, Claudio Martin Jonsson and Carla Beatriz Grespan Bottoli declare no conflict of interest.
This article contains studies with animals approved by Ethical Commission for the Use of Animals (CEUA) of the Embrapa Environment (Registration 007/17) (Jonsson et al. 2017).
- Anastassiades M, Lehotay SJ, Stajnbaher D, Schenck FJ (2003) Fast and easy multiresidue method employing acetonitrile extraction/partitioning and “dispersive solid-phase extraction” for the determination of pesticide residues in produce. J Assoc Off Anal Chem 86:412–431Google Scholar
- Branco LCC (2016) Farmacocinética e Depleção de resíduos do florfenicol em tambaqui (Colossoma macropomun). University of Campinas, Brazil, DissertationGoogle Scholar
- EC (2002) European Comission. Commission Regulation n° 657/2002 of 12 August 2002Google Scholar
- EC (2009) European Comission. Commission Regulation n° 37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin, https://ec.europa.eu/health/sites/health/files/files/eudralex/vol-5/reg_2010_37/reg_2010_37_en.pdf . Acessed 30 October 2018
- FAO (2018) Food and Agriculture Organization of the United Nations; the state of world fisheries and aquaculture, http://www.fao.org/3/i9540en/I9540EN.pdf. Acessed 30 October 2018
- Food Ingredients Brasil (2009) Proteínas do peixe, 8:23–32Google Scholar
- Gaikowski MP, Mushtaq M, Cassidy P, Meinertz JR, Schleis SM, Sweeney D, Endris RG (2010) Depletion of florfenicol amine, marker residue of florfenicol, from the edible fillet of tilapia (Oreochromis niloticus×O. niloticus and O. niloticus×O. aureus) following florfenicol administration in feed. Aquaculture 301:1–6CrossRefGoogle Scholar
- Jonsson CM, Hisano H, Paraiba LC (2017) Comissão de ética de uso de animais. Acúmulo do florfenicol e oxitetraciclina em filé de tambaquis, tilápias e pacus através da água e da ração medicadaGoogle Scholar
- MPA (2011) Ministério da Pesca e Aquicultura, Boletim estatístico da pesca e aquicultura. http://www.icmbio.gov.br/cepsul/images/stories/biblioteca/download/estatistica/est_2011_bol__bra.pdf. Acessed 30 October 2018
- Plumb DC (2004) Veterinary Drug Handbook, fifth edn. Iowa State PressGoogle Scholar
- MAPA (2011a) Ministério da Agricultura, Pecuária e Abastecimento. Instrução Normativa n° 24 de 01 de Junho de 2011. Acessed 30 October 2018Google Scholar
- MAPA (2011b) Ministério da Agricultura, Pecuária e Abastecimento. Guia de Validação e Controle de Qualidade Analítica para Fármacos em Produtos para Alimentação Animal e Medicamentos Veterinários, http://www.agricultura.gov.br/assuntos/laboratorios/arquivos-publicacoes-laboratorio/guia-de-validacao-controle-de-qualidade-analitica.pdf. Acessed 30 October 2018
- Martins ML, Primel EG, Caldas SS, Prestes OD, Adaime MB, Zanella R (2012) Microextração líquido-líquido dispersiva (DLLME): fundamentos e aplicações. Sci Chromatogr 4:35–51Google Scholar
- Phu TM, Scippo ML, Phuong NT, Tien CTK, Son CH, Dalsgaard A (2015) Withdrawal time for sulfamethoxazole and trimethoprim following treatment of striped catfish (Pangasianodon hypophthalmus) and hybrid red tilapia (Oreochromis mossambicus × Oreochromis niloticus). Aquacult 437:256–262CrossRefGoogle Scholar
- Tao Y, Zhu F, Chen D, Wei H, Pan Y, Wang X, Liu Z, Huang L, Wang Y, Yuan Z (2014) Evaluation of matrix solid-phase dispersion (MSPD) extraction for multi-fenicols determination in shrimp and fish by liquid chromatography–electrospray ionisation tandem mass spectrometry. Food Chem 150:500–506CrossRefGoogle Scholar
- Tsai WH, Chuang HY, Chen HH, Huang JJ, Chen HC, Cheng SH, Huang TC (2009) Application of dispersive liquid–liquid microextraction and dispersive micro-solid-phase extraction for the determination of quinolones in swine muscle by high-performance liquid chromatography with diode-array detection. Anal Chim Acta 656:56–62CrossRefGoogle Scholar