Determination of Florfenicol, Thiamfenicol and Chloramfenicol at Trace Levels in Animal Feed by HPLC–MS/MS

Abstract

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 µg/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.

Quantification and Analysis of Trace Levels of Phenicols in Feed by Liquid Chromatography–Mass Spectrometry

“…A second extraction with mixture of acetonitrile/ethyl acetate/ methanol was evaluated separately [52]. A third extraction used only ethyl acetate [2]. Ethyl acetate was better for the recovery of most of the remaining CAP and the rest of the pharmaceuticals as compared to previous two extraction methods.…”

“…Antibiotics are widely used for therapeutic and prophylactic purposes in human and veterinary medicine and also to promote growth and increase feed efficiencies in food producing animals [1]. However, abused use of antibiotics and their presence in the food of animal origin are of concern due to development of resistance in target pathogens against antibiotics, induced allergic reactions in some hypersensitive individuals, and potential compromise of the human intestinal and immune systems [2,3]. There is a diverse range of chemical substances with antimicrobial activity.…”

Simultaneous detection of three amphenicol antibiotics in shrimp and surface water samples by LC–MS/MS using two-antibodies-immobilized immunoaffinity clean-up technique

“…Various analytical methods including gas chromatography (GC) (Fodey et al, 2007;Zhang et al, 2006), GC-mass spectrometry (MS) (Ding et al, 2005;Shen et al, 2009), liquid chromatography (LC) (Samanidou et al, 2015;Xie et al, 2011), LC-MS/MS (Barreto et al, 2016;Gavilán et al, 2019;Hanna et al, 2018), enzyme-linked immunosorbent assays (ELISA) and lateral flow immunochromatographic assay (LFIA) (An et al, 2016;Byzova et al, 2010;Guo et al, 2015;Liu et al, 2014), etc. have been reported for the determination of CAP, FF and TAP in different matrices such as fish, shrimp, milk, egg, poultry, swine, bovine, feed and surface water (Guo et al, 2015;Zhang et al, 2006).…”

Oxytetracycline and Florfenicol Concentrations in Food-Additive Premixes Authorised for Broiler Chickens: Assessing Degree of Agreement with Manufacturers Labelling

“…This technique has been used by several researchers to identify unknown compounds, quantify known ones, and to know the structure and chemical properties of molecules [ 37 ]. The detection capability and soundness of this kind of instruments makes this the technique of choice for studies on the equivalence of chemical compounds that try to determine via spectroscopy the amount of antimicrobial drugs present in premixes [ 38 , 39 ].…”