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The use of veterinary medicinal products in food producing animals for a variety of purposes causes that their residues may be presented in edible tissues. As a result, in concern of public health, European Union Countries establish each year monitoring plans and they control the levels of harmful substances in food of animal origin. This paper presents survey of residues of veterinary medicinal products and coccidiostats in eggs for Poland and European Union in years 2007-2010. Despite the decrease in reported non-compliant results for coccidiostats, the numbers were still higher than those for veterinary medicines. The most often determined coccidiostats were: nicarbazin, dinitrocarbanilide, salinomycin and lasalocid, and the most often reported non-compliant results for veterinary medicines were: antimicrobials, enrofloxacin and doxycycline.
The method for confirmatory analysis of clazuril, decoquinate, diclazuril, halofuginone, lasalocid, maduramicin, monensin, narasin, niearbazin, robenidine, salinomycin, and semduramicin in eggs has been developed and validated. The samples were extracted with acetonitrile, purified on Oasis HLB SPE columns and analysed by liquid chromatography coupled with tandem mass spectrometry. The validation, performed according to the EU requirements, proved that the method enables the confirmation of coccidiostats in eggs at the levels required in the official control of residues (CCα in the range of 2.92-178 µg/kg, depending on the coccidiostat). The within-laboratory reproducibility (CV in the range of 6.1-29.3%) is also acceptable. The method was successfully verified in the proficiency test and implemented in the national residue control plan.
The experiment was performed to define the depletion of dinitrocarbanilide (DNC, marker of nicarbazin residue) and narasin residues in hen tissues and eggs after administration of Maxiban (nicarbazin and narasin, 80 mg kg⁻¹). A flock of 50 hens were kept in deep litter and fed coccidiostats for 14 d following 22 d of withdrawal. Randomly selected birds were slaughtered after the withdrawal of anticoccidial-containing feed. Samples of eggs, liver, and breast muscles were collected and analysed using validated LC-MS/MS method. The concentration of dinitrocarbanilide in the liver on day 0 of the withdrawal was 4,440 ±569 µg kg⁻¹ and quickly dropped to reach plateau level after 5 d. Long persistence of dinitrocarbanilide at plateau level and its presence in gastric contents during the withdrawal suggest the recycling of DNC with litter. The concentration of narasin in tissues and eggs was low even at the beginning of withdrawal period, which confirms the low probability of occurrence of narasin residues in food, even in the case of off-label use of this coccidiostat.
A multiresidue method (LC-MS/MS) for determination of wide range of anthelmintics was developed. The method covered benzimidazoles: albendazole (and metabolites), cambendazole, fenbendazol (and metabolites), flubendazole (and metabolites), mebendazole (and metabolites), oxibendazole, thiabendazole (and metabolites), triclabendazole (and metabolites); macrocyclic lactones: abamectin, doramectin, emamectin, eprinomectin, ivermectin, moxidectin; salicylanilides: closantel, ioxynil, nitroxynil, oxyclosamide, niclosamide, rafoxanid and others: clorsulon, derquantel, imidocarb, monepantel (and metabolites), morantel, praziquantel, and pyrantel. The method was used to examine the potential presence of anthelmintics in goat and sheep milk and dairy products from the Polish market. A total of 120 samples of milk, yoghurt, cottage cheese, cream cheese, and curd were analysed. None of the samples were found positive above CCa (1-10 µg/kg) except for one cottage cheese in which traces of albendazole sulfone were detected (5.2 µg/kg) and confirmed. The results of the study showed negligible anthelmintic residues in the goat and sheep milk and dairy products and confirm their good quality.
It is an old practice for food producers to colour their products to make them look more attractive to consumers. Also egg yolks are coloured to give them an appealing appearance. Owing to highly developed trade within the European Union countries, regulations for the usage of food dyes have been harmonized and the risks of harmful effects of some of these substances have been re-evaluated. In some countries of the European Union even dyes that have been approved as food additives are subjected to special surveillance or banned. Some additives are mutagenic and carcinogenic, and therefore their presence in food (including food of animal origin) is deemed unsafe. Illegal use of industrial dyes to colour egg yolks destined for human consumption has led to the development of accurate analytical methods for their detection. This paper includes, among others, a review of procedures applied for preparing samples for chromatographic analysis by methods published over the last ten years which are used to determine banned dyes and canthaxanthin in eggs. The health risk to consumers from the presence of industrial dyes in eggs makes it necessary to monitor their occurrence in eggs and egg products, as well as to include them in the Polish “National control programme for prohibited substances and the residues of chemical, biological, and veterinary medicinal products in live animals and animal products”.
A simple and short HPLC-UV screening method for the determination of residues for seven NSAIDs (carprofen, diclofenac, flunixin, meloxicam, phenylbutazone, tolfenamic acid, vedaprofen) and their three metabolites (4-methylaminoantipyrine, 5-hydroxyflunixin, oxyphenbutazone) in cow's milk has been developed. The sample preparation was based on liquid-liquid extraction with acetonitrile in the presence of sodium chloride. The separation of analytes was performed on a C18 column with a gradient of acetonitrile and the ammonium acetate buffer pH 5.0. UV-detector wavelength was programmed in order to improve sensitivity. The method was validated according to the CD/2002/657 criteria. For most analytes, relatively high recoveries were observed (76%-98%). Within-laboratory reproducibility levels were in the range of 3.6%-17.8% (CV, %). For phenylbutazone, oxyphenbutazone, and 4-methylaminoantipyrine recoveries were considerably lower (44%-68%) and reproducibility was up to 41.9%, which was probably caused by the instability of analytes. The robustness of the method for different fat content was successfully investigated. The method was verified by its use in the determination of meloxicam residues in milk samples obtained from meloxicam-treated cows. The obtained results confirm the usefulness of the developed method for the analysis of NSAIDs residues in milk.
A sensitive and reliable method has been developed and validated to determine residues of abamectin, doramectin, eprinomectin, ivermectin, and moxidectin in bovine milk. Isolation of the analytes from milk was performed with the use of liquid- liquid extraction with acetonitrile in the presence of sodium chloride. The extract was defatted with hexane and cleaned up using solid phase extraction (C8 cartridge) after forming ion pairs with triethylamine. The analytes were derivatized with N, N- dimethylformamide, acetic acid anhydride, and N-methylimidazole (100°C, 90 min). The derivatives were determined by reverse phase liquid chromatography with fluorescence detection (excitation and emission wavelength 365 nm and 475 nm, respectively). Recoveries of the lactones from milk samples fortified at 10-30 µg kg⁻¹ ranged from 52% to 80% with intra-laboratory reproducibility (CV) of 12.7%-22.8%. The critical concentrations (decision limit, CCα and detection capability, CCß) were in accordance with target limits. The method has been verified in the proficiency studies by EURL/CVL Berlin (all z-scores in the range of ±2). The method was transferred to routine laboratories, verified in inter-laboratory comparison and successfully applied in the National Residue Control Plan.
Mycotoxins are a wide group of compounds that often occur in food and feeds. These toxins negatively affect living organisms and may pose a risk for consumers’ health. Fish seem to be a particularly vulnerable group because negative effects can be observed at low levels of contamination, much lower than those considered harmful for other farmed animals. Mycotoxins may disturb cellular homeostasis by their influence on cells’ metabolism, causing DNA damage and organ lesions. Moreover, they are capable of immunomodulation, which may increase the occurrence of fish diseases caused by bacteria and other pathogens. Furthermore, these substances often have a negative impact on the growth rate of fish, which may cause economic losses to farmers. Although mycotoxins are commonly found in feeds, their ability to bioaccumulate in fish seems to be marginal. Therefore, according to the available data, fish may not be considered as the main source of mycotoxins for humans. Since there are no legal limits on the amount of mycotoxins present in fish feeds, it is necessary to constantly monitor the levels of mycotoxins in fish feeds and to investigate the influence of these substances on fish health.
A multiresidue method for simultaneous determination of 10 dyes unauthorised for the use in laying hens was developed (Sudan I, Sudan II, Sudan III, Sudan IV, Sudan Red G, Sudan Orange G, Sudan Red 7B, Para-Red, Toluidine Red, Citrus Red). The dyes were extracted using liquid-liquid extraction with acetonitrile in the presence of anhydrous sodium sulphate, and cleaned using zirconium coated silica cartridges. After dilution with acetonitrile saturated with hexane:DMSO (8:2), samples were analysed using LC-MS/MS system with acetonitrile (A) and 0.1% formic acid (B) as a mobile phase in a gradient mode and C₁₈ analytical column. The method was validated according to the requirements described in the Commission Decision 2002/657/EC: linearity (r ≥ 0.998), precision: repeatability (1.94%-10.02%), and within-laboratory reproducibility (4.66%-8.89%), recovery (97%-105%), decision limit CCα (5.33-6.50 µg/kg), and detection capability CCß (6.18-7.50 µg/kg) were calculated. The developed method fulfilled all performance criteria and can be used in the official survey of dyes residues in food of animal origin.
The method for quantitative determination, of flunixin and its metabolite 5-hydroxyflunixin in bovine plasma has been developed and validated. Plasma samples were extracted with acetonitrile and analysed by liquid chromatography with ultraviolet detection. Extraction efficiency was tested and optimum pH conditions were established. Limit of the detection for both compounds was 0.03 µg/mL, limits of quantitation of flunixin and 5- hydroxyflunixin were 0.06 and 0.05 µg/mL, respectively. Both substances were determined with an accuracy below 12% (CV, %) and overall recovery in the range of 63-85%. The method was verified by the analyses of plasma samples from cows treated with flunixin meglumine.
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