Determination and confirmation of metronidazole, dimetridazole, ronidazole and their metabolites in bovine muscle by LC-MS/MS

Abstract

Nitroimidazoles are a class of veterinary drugs used for the treatment and prevention of certain bacterial and protozoal diseases in poultry, swine dysentery and genital trichomoniasis in cattle. Since the safety assessment of nitroimidazoles showed them to be genotoxic, carcinogenic and mutagenic, a number of nitroimidazoles have been banned for therapeutic purposes in different countries. Moreover, nitroimidazoles have also been extensively used as growth promoters in food-producing animals. Due to their efficacious improvement in meat production and feed conversion, deliberate use still exists. Therefore, the illegal use of nitroimidazoles in animal husbandry must be monitored. A sensitive method based on LC-MS/MS for the simultaneous determination and confirmation of five banned nitroimidazole drugs including metronidazole, ronidazole, dimetridazole, metronidazole-OH (metabolite of metronidazole), and 2-hydroxymethyl-1-methyl-5-nitroimidazole (metabolite of ronidazole and dimetridazole) in bovine muscle, using ronidazole-d3 as an internal standard, was developed and validated. After extraction with ethyl acetate and evaporation, the nitroimidazoles were reconstituted in petroleum ether and purified, and LC-MS/MS analysis was performed. The method was validated according to Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC). Parameters such as decision limit (CCα), detection capability (CCβ), precision, accuracy, uncertaincy and ruggedness were determined. Average accuracy of the five nitroimidazoles from bovine muscle fortified at 5 levels (0.5, 1.0, 1.5, 2.0 and 2.5 μg kg(-1)) ranged from 96% to 103%. The calculated CCα ranged from 0.0 to 0.17 μg kg(-1); CCβ ranged from 0.08 to 0.41 μg kg(-1). A complete statistical analysis was performed and the results indicate that the method is robust when subjected to day-to-day analytical variations.

Metronidazole pharmacokinetics during rapid growth in turkeys – relation to changes in haemodynamics and drug metabolism

“…Despite extensive use, MTZ PK studies in turkeys are scarce in the scientific literature. As the ban for the use of MTZ in poultry left little alternative for the treatment of histomoniasis, the illicit use of the drug is still reported (Granja et al, 2013).…”

Determination of 5-nitroimidazole residues in milk by capillary electrochromatography with packed C18 silica beds

“…Liquid chromatography (LC) with UV [8][9][10] or mass spectrometry (MS) [11][12][13][14] detection is the most employed technique. Also, gas chromatography (GC) coupled to MS [15,16] or capillary electrophoresis (CE) with UV detection [17,18] have been proposed.…”

Chemical and physical hazards in the Dutch poultry meat chain

“…5-nitroimidazoles are classified based on their nitro functional group location. Some examples include DMZ, MNZ, and RNZ (Granja et al, 2013;Trevisani et al, 2014;Wang, 2001;Xia et al, 2006). 5-nitroimidazoles have been used to treat and prevent some bacterial and protozoal diseases in poultry like histomoniasis and coccidiosis (Granja et al, 2013;Hurtaud-Pessel, Delepine, & Laurentie, 2000;Wang, 2001;Xia et al, 2006).…”

“…Some examples include DMZ, MNZ, and RNZ (Granja et al, 2013;Trevisani et al, 2014;Wang, 2001;Xia et al, 2006). 5-nitroimidazoles have been used to treat and prevent some bacterial and protozoal diseases in poultry like histomoniasis and coccidiosis (Granja et al, 2013;Hurtaud-Pessel, Delepine, & Laurentie, 2000;Wang, 2001;Xia et al, 2006). In particular, 5-nitroimidazoles are reported to be rapidly metabolized with the main metabolites resulting from the oxidation of the side chains at the C2 position on the imidazole ring; the resulting metabolites may have a similar mutagenic potential as the parent compound (Hurtaud-Pessel et al, 2000).…”

“…(EFSA Panels on Biological Hazards (BIOHAZ) et al, 2012). Hence, monitoring remains imperative (Granja et al, 2013;Reeves, 2011) as nitroimidazoles have been reported as effective growth promoters in meat production and feed conversion (Granja et al, 2013) (particularly the beta-agonist cimaterol). None of them were positive for these compounds.…”