{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T20:01:32Z","timestamp":1760385692304,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2019,5,7]],"date-time":"2019-05-07T00:00:00Z","timestamp":1557187200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100010801","name":"Xunta de Galicia","doi-asserted-by":"publisher","award":["FEADER 2018\/001B","ED431C 2018\/05"],"award-info":[{"award-number":["FEADER 2018\/001B","ED431C 2018\/05"]}],"id":[{"id":"10.13039\/501100010801","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Administration of florfenicol and thiamfenicol through medicated feed is permitted within the European Union, always following veterinary prescription and respecting the withdrawal periods. However, the presence of low levels of florfenicol, thiamfenicol, and chloramfenicol in non-target feed is prohibited. Since cross-contamination can occur during the production of medicated feed and according to Annex II of the European Regulation 2019\/4\/EC, the control of residue levels of florfenicol and thiamfenicol in non-target feed should be monitored and avoided. Based on all the above, a sensitive and reliable method using liquid chromatography tandem mass spectrometry was developed for the simultaneous detection of chloramfenicol, florfenicol, and thiamfenicol at trace levels in animal feed. Analytes were extracted from minced feed with ethyl acetate. Then, the ethyl acetate was evaporated, the residue was resuspended in Milli-Q water and the extract filtered. The method was in-house validated at carryover levels, with concentration ranging from 100 to 1000 \u00b5g\/kg. The validation was conducted following the European Commission Decision 2002\/657\/EC and all performance characteristics were successfully satisfied. The capability of the method to detect amfenicols at lower levels than any prior perspective regulation literature guarantees its applicability in official control activities. The developed method has been applied to non-compliant feed samples with satisfactory results.<\/jats:p>","DOI":"10.3390\/antibiotics8020059","type":"journal-article","created":{"date-parts":[[2019,5,9]],"date-time":"2019-05-09T11:22:35Z","timestamp":1557400955000},"page":"59","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Determination of Florfenicol, Thiamfenicol and Chloramfenicol at Trace Levels in Animal Feed by HPLC\u2013MS\/MS"],"prefix":"10.3390","volume":"8","author":[{"given":"Rosa Elvira","family":"Gavil\u00e1n","sequence":"first","affiliation":[{"name":"Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Veterinary Medicine, University of Santiago de Compostela, 27002 Lugo, Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2141-3002","authenticated-orcid":false,"given":"Carolina","family":"Nebot","sequence":"additional","affiliation":[{"name":"Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Veterinary Medicine, University of Santiago de Compostela, 27002 Lugo, Spain"}]},{"given":"Ewelina","family":"Patyra","sequence":"additional","affiliation":[{"name":"Department of Hygiene of Animal Feedingstuffs, National Veterinary Research Institute, 24\u2013100 Pulawy, Poland"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9305-0151","authenticated-orcid":false,"given":"Beatriz","family":"Vazquez","sequence":"additional","affiliation":[{"name":"Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Veterinary Medicine, University of Santiago de Compostela, 27002 Lugo, Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7992-1491","authenticated-orcid":false,"given":"Jose Manuel","family":"Miranda","sequence":"additional","affiliation":[{"name":"Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Veterinary Medicine, University of Santiago de Compostela, 27002 Lugo, Spain"}]},{"given":"Alberto","family":"Cepeda","sequence":"additional","affiliation":[{"name":"Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Veterinary Medicine, University of Santiago de Compostela, 27002 Lugo, Spain"}]}],"member":"1968","published-online":{"date-parts":[[2019,5,7]]},"reference":[{"key":"ref_1","unstructured":"(2017, January 29). ESVAC (2015) Fifth ESVAC Report\u2014Sales of Veterinary Antimicrobial Agents in 26 EU\/EEA Countries in 2013. Available online: www.ema.europa.eu\/docs\/en_GB\/document_library\/Report\/2015\/10\/WC500195687.pdf."},{"key":"#cr-split#-ref_2.1","unstructured":"Europe Comission (2010). Regulation 37\/2010. Commission Regulation"},{"key":"#cr-split#-ref_2.2","unstructured":"(EU) No 37\/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin. Off. J. Eur. Union, 15, 1-72."},{"key":"ref_3","unstructured":"Europe Comission (2019). Regulation 2019\/4. Commission Regulation (EU) 2019\/4 of the European Parliament and of the Council of 11 December 2018 on the manufacture, placing on the market and use of medicated feed, amending Regulation (EC) No 183\/2005 of the European Parliament and of the Council and repealing Council Directive 90\/167\/EEC (Text with EEA relevance). Off. J. Eur. Union, 4, 1\u201323."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1002\/bmc.3615","article-title":"Simplifying sample preparation using fabric phase sorptive extraction technique for the determination of benzodiazepines in blood serum by high performance liquid chromatography","volume":"30","author":"Samanidou","year":"2016","journal-title":"Biomed. Chromatogr."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2434","DOI":"10.1007\/s12161-016-0436-3","article-title":"Development and validation of a sensitive indirect competitive enzyme-linked immunosorbent assay for the screening of florfenicol and Tiamfenicol in edible animal tissue and feed","volume":"9","author":"An","year":"2016","journal-title":"Food Anal. Method"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1016\/j.jpba.2010.01.030","article-title":"On-line molecular imprinted solid-phase extraction flow-injection fluorescence sensor for determination of florfenicol in animal tissues","volume":"52","author":"Ge","year":"2010","journal-title":"J. Pharm. Biomed."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"7","DOI":"10.5740\/jaoacint.12-181","article-title":"Determination of florfenicol in fish feeds at high inclusion rates by HPLC-UV","volume":"96","author":"Hayes","year":"2013","journal-title":"J. AOAC Int."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1016\/j.jpba.2005.05.032","article-title":"Comparative evaluation between capillary electrophoresis and high-performance liquid chromatography for the analysis of florfenicol in plasma","volume":"39","author":"Kowalski","year":"2005","journal-title":"J. Pharm. Biomed."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/j.talanta.2015.12.045","article-title":"Determination of chloramfenicol, Tiamfenicol and florfenicol in milk and honey using modified QuEChERS extraction coupled with polymeric monolith-based capillary liquid chromatography tandem mass spectrometry","volume":"150","author":"Liu","year":"2016","journal-title":"Talanta"},{"key":"ref_10","unstructured":"Europe Comission (2002). Comission Decission 2002\/657\/EC. Commission Decision of 12 August 2002 implementing Council Directive 96\/23\/EC concerning the performance of analytical methods and the interpretation of results. Off. J. Eur. Union, 221, 8\u201336."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1093\/jaoac\/86.3.510","article-title":"Simultaneous determination of residues of chloramfenicol, tiamfenicol, florfenicol, and florfenicol amine in farmed aquatic species by liquid chromatography\/mass spectrometry","volume":"86","author":"Potter","year":"2003","journal-title":"J. AOAC Int."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.chroma.2016.04.024","article-title":"Determination of chloramfenicol, Tiamfenicol, florfenicol and florfenicol amine in poultry, swine, bovine and fish by liquid chromatography-tandem mass spectrometry","volume":"1449","author":"Barreto","year":"2016","journal-title":"J. Chromatogr. A"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.jchromb.2016.08.014","article-title":"Qualitative and quantitative drug residue analyses: Florfenicol in white-tailed deer (Odocoileus virginianus) and supermarket meat by liquid chromatography tandem-mass spectrometry","volume":"1033","author":"Anderson","year":"2016","journal-title":"J. Chromatogr. B"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1016\/j.foodchem.2018.07.045","article-title":"Quantitative analysis of chloramfenicol, tiamfenicol, florfenicol and florfenicol amine in eggs via liquid chromatography-electrospray ionization tandem mass spectrometry","volume":"269","author":"Xie","year":"2018","journal-title":"Food Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"621","DOI":"10.2478\/bvip-2014-0095","article-title":"Amfenicols stability in medicated feed\u2013development and validation of liquid chromatography method","volume":"58","author":"Pietro","year":"2014","journal-title":"Bull. Vet. Inst. Pulawy"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.jchromb.2017.09.045","article-title":"Development of a high-performance liquid chromatography method for the determination of florfenicol in animal feedstuffs","volume":"1068","author":"Yang","year":"2017","journal-title":"J. Chromatogr. B"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.foodchem.2014.03.086","article-title":"Qualitative screening of 116 veterinary drugs in feed by liquid chromatography\u2013high resolution mass spectrometry: Potential application to quantitative analysis","volume":"160","author":"Boix","year":"2014","journal-title":"Food Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1016\/j.foodcont.2014.09.040","article-title":"Rapid multiresidue and multi-class screening for antibiotics and benzimidazoles in feed by ultra-high performance liquid chromatography coupled to tandem mass spectrometry","volume":"50","author":"Robert","year":"2015","journal-title":"Food Control"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6543","DOI":"10.1007\/s00216-013-7060-5","article-title":"Wide-scope analysis of veterinary drug and pesticide residues in animal feed by liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry","volume":"405","author":"Vidal","year":"2013","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1637","DOI":"10.1080\/19440049.2015.1023742","article-title":"Analysis of veterinary drug and pesticide residues in animal feed by high-resolution mass spectrometry: Comparison between time-of-flight and Orbitrap","volume":"32","year":"2015","journal-title":"Food Addit. Contam. A"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.talanta.2015.11.032","article-title":"Target analysis and retrospective screening of veterinary drugs, ergot alkaloids, plant toxins and other undesirable substances in feed using liquid chromatography\u2013high resolution mass spectrometry","volume":"149","author":"Pastor","year":"2016","journal-title":"Talanta"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"6394","DOI":"10.1016\/j.chroma.2010.08.024","article-title":"Development of a multi-class method for the quantification of veterinary drug residues in feedingstuffs by liquid chromatography-tandem mass spectrometry","volume":"1217","author":"Boscher","year":"2010","journal-title":"J. Chromatogr. A"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1016\/j.foodchem.2015.10.076","article-title":"Multiresidue method for the simultaneous determination of veterinary medicinal products, feed additives and illegal dyes in eggs using liquid chromatography\u2013tandem mass spectrometry","volume":"197","author":"Piatkowska","year":"2016","journal-title":"Food Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/j.jchromb.2008.09.035","article-title":"Simultaneous determination and confirmation of chloramfenicol, tiamfenicol, florfenicol and florfenicol amine in chicken muscle by liquid chromatography\u2013tandem mass spectrometry","volume":"875","author":"Zhang","year":"2008","journal-title":"J. Chromatogr. B"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.1080\/19440049.2010.489579","article-title":"Rapid multi-class multi-residue method for the confirmation of chloramfenicol and eleven nitroimidazoles in milk and honey by liquid chromatography-tandem mass spectrometry (LC-MS)","volume":"27","author":"Cronly","year":"2010","journal-title":"Food Addit. Contam."},{"key":"ref_26","first-page":"125","article-title":"Simultaneous determination of residues of chloramfenicol and florfenicol in animal feed by liquid chromatography tandem mass spectrometry","volume":"8","author":"Shinoda","year":"2011","journal-title":"J. Resid. Sci. Technol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1481","DOI":"10.2134\/jeq2012.0014","article-title":"Occurrence of ciprofloxacin, enrofloxacin, and florfenicol in animal wastewater and water resources","volume":"41","author":"Wei","year":"2012","journal-title":"J. Environ. Qual."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.chroma.2015.09.047","article-title":"Development of a subcritical water extraction approach for trace analysis of chloramfenicol, tiamfenicol, florfenicol, and florfenicol amine in poultry tissues","volume":"1418","author":"Xiao","year":"2015","journal-title":"J. Chromatogr. A"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1093\/jaoac\/92.4.1225","article-title":"Simultaneous determination of residual tiamfenicol and florfenicol in foods of animal origin by HPLC\/electrospray ionization-MS\/MS","volume":"92","author":"Chou","year":"2009","journal-title":"J. AOAC Int."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/S1872-2059(06)60002-3","article-title":"Simultaneous determination of chloramfenicol, tiamfenicol, and florfenicol residues in animal tissues by gas chromatography\/mass spectrometry","volume":"24","author":"Peng","year":"2006","journal-title":"Chin. J. Chromatogr."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.jchromb.2009.10.002","article-title":"Simultaneous determination of thiamfenicol, florfenicol and florfenicol amine in swine muscle by liquid chromatography\u2013tandem mass spectrometry with immunoaffinity chromatography clean-up","volume":"878","author":"Luo","year":"2010","journal-title":"J. Chromatogr. B"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1016\/j.idairyj.2011.09.001","article-title":"Development of a multi-class method for the identification and quantification of residues of antibiotics, coccidiostats and corticosteroids in milk by liquid chromatography\u2013tandem mass spectrometry","volume":"22","author":"Nebot","year":"2012","journal-title":"Int. Dairy J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1016\/j.foodcont.2014.05.042","article-title":"Monitoring the presence of residues of tetracyclines in baby food samples by HPLC-MS\/MS","volume":"46","author":"Nebot","year":"2014","journal-title":"Food Control"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Gavil\u00e1n, R., Nebot, C., Miranda, J., Mart\u00edn-G\u00f3mez, Y., V\u00e1zquez-Belda, B., Franco, C., and Cepeda, A. (2016). Analysis of tetracyclines in medicated feed for food animal production by HPLC-MS\/MS. Antibiotics, 5.","DOI":"10.3390\/antibiotics5010001"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1080\/19440049.2018.1466061","article-title":"Simultaneous analysis of coccidiostats and sulphonamides in non-target feed by HPLC-MS\/MS and validation following the Commission Decision 2002\/657\/EC","volume":"35","author":"Nebot","year":"2018","journal-title":"Food Addit. Contam. A"}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/8\/2\/59\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:49:52Z","timestamp":1760186992000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/8\/2\/59"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,5,7]]},"references-count":36,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2019,6]]}},"alternative-id":["antibiotics8020059"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics8020059","relation":{},"ISSN":["2079-6382"],"issn-type":[{"type":"electronic","value":"2079-6382"}],"subject":[],"published":{"date-parts":[[2019,5,7]]}}}