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Ursula Vincent - One of the best experts on this subject based on the ideXlab platform.
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Screening methods and recent developments in the detection of anticoccidials
Analytical and Bioanalytical Chemistry, 2013Co-Authors: Anne-catherine Huet, Ursula Vincent, Monique Bienenmann-ploum, Philippe DelahautAbstract:This article presents a review of the current trends in the analysis of Coccidiostats in various matrices, focusing principally on screening and rapid methods. Coccidiosis is an infectious disease having a high negative impact on the animal industry. Drugs are therefore necessary to prevent and/or to combat this disease. However, it is also of crucial importance that these veterinary drugs do not enter the human food chain. European legislation has therefore established the boundaries for the use of coccidiosats and has also addressed the unavoidable problem of cross-contamination of the feed, mainly caused by the use of the same production lines. Consequently there is a need for analytical methods and/or analytical strategies for the monitoring and control of the residues of anticoccidials, both in feed and in the resulting matrices for human consumption. In the frame of the European collaborative project CON ff IDENCE, such attempts to establish the required analytical tools were made, which required beforehand a review of the state of the art in this domain. Aiming at this objective, in this review we consider the most interesting publications since 2000. In essence, both a rapid approach with mainly immunoassays and chromatographic methods were developed. To date, the obstacle to routine use of the first approach has been its inability to detect more than two compounds simultaneously, but recent developments in flow cytometry have made it possible to detect six Coccidiostats at once. On the other hand, an increasingly popular approach for detecting multiple Coccidiostats simultaneously is liquid chromatography coupled with tandem mass spectrometry. There remains a need to adapt these analytical methods to legislative requirements.
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Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six Coccidiostats in feed and eggs
Analytical and Bioanalytical Chemistry, 2012Co-Authors: Monique E. Bienenmann-ploum, Terence L. Fodey, Philippe Delahaut, Ursula Vincent, Anne-catherine Huet, Katrina Campbell, Willem Haasnoot, Christopher T. Elliott, Michel W. F. NielenAbstract:Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these Coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority Coccidiostats. Different Coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation ( r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4′-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 μg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 μg/kg, respectively. Figure
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Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six Coccidiostats in feed and eggs
Analytical and Bioanalytical Chemistry, 2012Co-Authors: Monique E. Bienenmann-ploum, Terence L. Fodey, Philippe Delahaut, Ursula Vincent, Anne-catherine Huet, Katrina Campbell, Willem Haasnoot, Christopher T. Elliott, Michel W. F. NielenAbstract:Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these Coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority Coccidiostats. Different Coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation (r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4′-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 μg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 μg/kg, respectively.
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Determination of ionophore Coccidiostats in feedingstuffs by liquid chromatography–tandem mass spectrometry: Part I. Application to targeted feed
Journal of Pharmaceutical and Biomedical Analysis, 2008Co-Authors: Ursula Vincent, Mostafa Chedin, Sulhattin Yasar, Christoph Von HolstAbstract:Abstract A new and fit to purpose multi-analyte method for the determination of six Coccidiostats (monensin A, salinomycin, narasin composed of its principal component narasin A and its minor component narasin I, lasalocid, semduramicin and maduramicin) in poultry and cattle compound feed by liquid chromatography tandem mass spectrometry (LC–MS/MS) has been developed and in-house validated. The concentration level of the target analytes at which the validation experiments have been carried out varied between 1 and 9 mg kg −1 . The method developed involved a simple extraction of the Coccidiostats from the feed samples followed by a clean-up by solid-phase extraction prior to chromatographic analysis. The analytes were quantified either by matrix-matched standards or by the standard addition technique, obtaining the following performance profile of the method for the various analyte/matrix combinations. When quantifying against matrix-matched standards, the concentration independent intermediate precision expressed in terms of relative percentage standard deviation varied between 4 and 10% and the relative percentage recovery rates ranged from 86 to 120%, depending on the target analyte and matrix. When using the standard addition technique, the corresponding values for the intermediate precision varied between 2 and 8% and the relative percentage recovery rate ranged from 73 to 115%. The limit of detection (LOD) and limit of quantification (LOQ) were different for the various analyte/matrix combinations but were in all cases below 0.014 and 0.046 mg kg −1 , respectively. Based on the obtained method performance characteristics, the method is considered suitable for the determination of ionophore Coccidiostats in target feed. The main field of application of the validated method is to enforce European legislation regarding the authorisation of Coccidiostats, focusing on the measurement at the authorised levels and at low level in feed during the withdrawal period at which the Coccidiostats must not be added to the feed. Overall, the method proposed appears to be appropriate as a confirmatory method for the monitoring of these six ionophore Coccidiostats and can therefore be considered as complementary to the official HPLC-UV methods.
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Determination of ionophore Coccidiostats in feedingstuffs by liquid chromatography-tandem mass spectrometry Part I. Application to targeted feed.
Journal of pharmaceutical and biomedical analysis, 2008Co-Authors: Ursula Vincent, Mostafa Chedin, Sulhattin Yasar, Christoph Von HolstAbstract:A new and fit to purpose multi-analyte method for the determination of six Coccidiostats (monensin A, salinomycin, narasin composed of its principal component narasin A and its minor component narasin I, lasalocid, semduramicin and maduramicin) in poultry and cattle compound feed by liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed and in-house validated. The concentration level of the target analytes at which the validation experiments have been carried out varied between 1 and 9 mg kg(-1). The method developed involved a simple extraction of the Coccidiostats from the feed samples followed by a clean-up by solid-phase extraction prior to chromatographic analysis. The analytes were quantified either by matrix-matched standards or by the standard addition technique, obtaining the following performance profile of the method for the various analyte/matrix combinations. When quantifying against matrix-matched standards, the concentration independent intermediate precision expressed in terms of relative percentage standard deviation varied between 4 and 10% and the relative percentage recovery rates ranged from 86 to 120%, depending on the target analyte and matrix. When using the standard addition technique, the corresponding values for the intermediate precision varied between 2 and 8% and the relative percentage recovery rate ranged from 73 to 115%. The limit of detection (LOD) and limit of quantification (LOQ) were different for the various analyte/matrix combinations but were in all cases below 0.014 and 0.046 mg kg(-1), respectively. Based on the obtained method performance characteristics, the method is considered suitable for the determination of ionophore Coccidiostats in target feed. The main field of application of the validated method is to enforce European legislation regarding the authorisation of Coccidiostats, focusing on the measurement at the authorised levels and at low level in feed during the withdrawal period at which the Coccidiostats must not be added to the feed. Overall, the method proposed appears to be appropriate as a confirmatory method for the monitoring of these six ionophore Coccidiostats and can therefore be considered as complementary to the official HPLC-UV methods.
Philippe Delahaut - One of the best experts on this subject based on the ideXlab platform.
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A review of Coccidiostats and the analysis of their residues in meat and other food
Meat Science, 2014Co-Authors: Lesa Clarke, Terence L. Fodey, Steven R. H. Crooks, Mary Moloney, John O’mahony, Philippe Delahaut, Richard O'kennedy, Martin DanaherAbstract:Coccidiostats are used in the control of protozoan infections in different food producing animals. They are most widely used as feed additives in intensively reared species such as pigs and poultry to maintain animal health and in some cases enhance feed conversion. However, a number of these drugs are used in the control of infections in beef and lamb production. Coccidiostat residues have been frequently reported in meat and eggs in a number of countries since the late 1990s. This has prompted increased research and surveillance of Coccidiostat residues in food. This paper reviews the various Coccidiostat agents used in animal production, including their chemical properties, mode of action and activity. Legislation concerning Coccidiostats, limits for residues in food, monitoring and occurrence of residues in food is discussed. Methods for residue determination in food, including screening and physicochemical methods are discussed in depth. The paper concludes with a synopsis of the current state of Coccidiostat residue analysis and future perspectives.
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Screening methods and recent developments in the detection of anticoccidials
Analytical and Bioanalytical Chemistry, 2013Co-Authors: Anne-catherine Huet, Ursula Vincent, Monique Bienenmann-ploum, Philippe DelahautAbstract:This article presents a review of the current trends in the analysis of Coccidiostats in various matrices, focusing principally on screening and rapid methods. Coccidiosis is an infectious disease having a high negative impact on the animal industry. Drugs are therefore necessary to prevent and/or to combat this disease. However, it is also of crucial importance that these veterinary drugs do not enter the human food chain. European legislation has therefore established the boundaries for the use of coccidiosats and has also addressed the unavoidable problem of cross-contamination of the feed, mainly caused by the use of the same production lines. Consequently there is a need for analytical methods and/or analytical strategies for the monitoring and control of the residues of anticoccidials, both in feed and in the resulting matrices for human consumption. In the frame of the European collaborative project CON ff IDENCE, such attempts to establish the required analytical tools were made, which required beforehand a review of the state of the art in this domain. Aiming at this objective, in this review we consider the most interesting publications since 2000. In essence, both a rapid approach with mainly immunoassays and chromatographic methods were developed. To date, the obstacle to routine use of the first approach has been its inability to detect more than two compounds simultaneously, but recent developments in flow cytometry have made it possible to detect six Coccidiostats at once. On the other hand, an increasingly popular approach for detecting multiple Coccidiostats simultaneously is liquid chromatography coupled with tandem mass spectrometry. There remains a need to adapt these analytical methods to legislative requirements.
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Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six Coccidiostats in feed and eggs
Analytical and Bioanalytical Chemistry, 2012Co-Authors: Monique E. Bienenmann-ploum, Terence L. Fodey, Philippe Delahaut, Ursula Vincent, Anne-catherine Huet, Katrina Campbell, Willem Haasnoot, Christopher T. Elliott, Michel W. F. NielenAbstract:Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these Coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority Coccidiostats. Different Coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation ( r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4′-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 μg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 μg/kg, respectively. Figure
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Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six Coccidiostats in feed and eggs
Analytical and Bioanalytical Chemistry, 2012Co-Authors: Monique E. Bienenmann-ploum, Terence L. Fodey, Philippe Delahaut, Ursula Vincent, Anne-catherine Huet, Katrina Campbell, Willem Haasnoot, Christopher T. Elliott, Michel W. F. NielenAbstract:Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these Coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority Coccidiostats. Different Coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation (r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4′-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 μg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 μg/kg, respectively.
Piotr Jedziniak - One of the best experts on this subject based on the ideXlab platform.
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CONFIRMATORY METHOD FOR DETERMINATION OF CoccidiostatS IN EGGS
Bulletin of The Veterinary Institute in Pulawy, 2020Co-Authors: Małgorzata Olejnik, Teresa Szprengier-juszkiewicz, Piotr JedziniakAbstract:The method for confirmatory analysis of clazuril, decoquinate, diclazuril, halofuginone, lasalocid, maduramicin, monensin, narasin, nicarbazin, 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.
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Determination of fifteen Coccidiostats in feed at carry-over levels using liquid chromatography-mass spectrometry.
Journal of Pharmaceutical and Biomedical Analysis, 2015Co-Authors: Konrad Pietruk, Małgorzata Olejnik, Piotr Jedziniak, Teresa Szprengier-juszkiewiczAbstract:Abstract A multi-residue method has been developed and validated for the simultaneous determination of authorized (decoquinate, diclazuril, halofuginone, lasalocid, maduramicin, monensin, narasin, nicarbazin, robenidine, salinomycin and semduramicin) and non-authorized (amprolium, clopidol, ethopabate and toltrazuril) Coccidiostats in animal feed. Feed samples were extracted with basic followed by acidified solution in methanol and, after centrifugation, were injected directly into LC–MS/MS system. Detection was performed in selected reaction monitoring mode with both positive and negative electrospray ionization. The time efficient validation experiment has verified the robustness of a method in different types of feed and on two separate LC–MS/MS instruments. The comparison of different quantification methods demonstrated that, against expectations, the standard addition did not prove better in comparison with matrix-matched calibration curve. Although the sample preparation was very easy, the observed matrix effects were not significant for the most part but they could explain the problems with the quantification of some Coccidiostats.
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comparison of different sample preparation procedures for multiclass determination of selected veterinary drug Coccidiostat and insecticide residues in eggs by liquid chromatography tandem mass spectrometry
Analytical Methods, 2014Co-Authors: M Piatkowska, Piotr Jedziniak, J ZmudzkiAbstract:This paper presents the evaluation of extraction techniques and sample clean-up procedure for the simultaneous determination of residues of veterinary drugs (sulphonamides, fluoroquinolones, tetracyclines, macrolides, β-lactams, nitroimidazoles, benzimidazoles, amphenicols, lincosamides and pleuromutilins), Coccidiostats and insecticides in fresh egg samples. The study utilises the liquid chromatography-tandem mass spectrometry technique to obtain suitable analytical parameters for the screening and confirmatory method. The results of egg samples analysed by 8 different sample preparation schemes were compared by using the mean recoveries. The recoveries of analytes showed that both SPE and modified QuEChERS have not been sufficient for the extraction of all the analytes in the expected range of 70–120%. Additional cleaning with dispersive sorbents prolonged the time taken by the sample preparation steps and provided slightly different final extract clarity results. The most demanding analytes to isolate from the samples were Coccidiostats, while sulphonamides were the group of analytes for which all the proposed sample clean-up procedures were suitable. As the chosen method, extraction with 0.1% formic acid in acetonitrile–water (8 : 2) and clean-up with HybridSPE can be applied as a sample preparation step for the screening and confirmatory method in official laboratories.
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In-house quality control material of nicarbazin and narasin in eggs: preparation and inter-laboratory evaluation
Accreditation and Quality Assurance, 2013Co-Authors: Małgorzata Olejnik, Teresa Szprengier-juszkiewicz, Piotr Jedziniak, Jan ŻmudzkiAbstract:Coccidiostats are a group of pharmacologically active substances widely used in veterinary practice. Their residues are detected relatively often in poultry tissues and egg samples analyzed as part of official residue control programs in the European Union. Therefore, accuracy of quantitative results needs to be monitored through internal and external quality control studies. In addition, the use of materials containing incurred residues would be welcome to for ongoing monitoring of the method accuracy. Unfortunately, in the field of veterinary drug residues, certified reference materials are often unavailable. Therefore, in-house quality control material of incurred lyophilized eggs containing narasin and nicarbazin has been produced and characterized. The eggs originated from hens receiving feed with Coccidiostat premix Maxiban were mixed to obtain presumed concentrations of residues and freeze-dried. Homogeneity of the material was verified by the duplicate analysis of ten random samples, and the results proved that the between samples variation was negligible in comparison with the method repeatability. No measurable loss of analytes was observed within 1 year; the slope of the regression line of the results of stability measurements was not significantly different from zero. The assigned values were expressed as medians of the results of inter-laboratory comparison performed in four different European laboratories; the uncertainty of the material was estimated, taken into consideration all above tests, resulting in (14.4 ± 2.53) µg/kg for nicarbazin and (7.91 ± 1.52) µg/kg for narasin.
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Residues of veterinary medicinal products and Coccidiostats in eggs - causes, control and results of surveillance program in Poland.
Polish Journal of Veterinary Sciences, 2012Co-Authors: Marta Piątkowska, Piotr Jedziniak, Jan ŻmudzkiAbstract: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.
Anne-catherine Huet - One of the best experts on this subject based on the ideXlab platform.
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Screening methods and recent developments in the detection of anticoccidials
Analytical and Bioanalytical Chemistry, 2013Co-Authors: Anne-catherine Huet, Ursula Vincent, Monique Bienenmann-ploum, Philippe DelahautAbstract:This article presents a review of the current trends in the analysis of Coccidiostats in various matrices, focusing principally on screening and rapid methods. Coccidiosis is an infectious disease having a high negative impact on the animal industry. Drugs are therefore necessary to prevent and/or to combat this disease. However, it is also of crucial importance that these veterinary drugs do not enter the human food chain. European legislation has therefore established the boundaries for the use of coccidiosats and has also addressed the unavoidable problem of cross-contamination of the feed, mainly caused by the use of the same production lines. Consequently there is a need for analytical methods and/or analytical strategies for the monitoring and control of the residues of anticoccidials, both in feed and in the resulting matrices for human consumption. In the frame of the European collaborative project CON ff IDENCE, such attempts to establish the required analytical tools were made, which required beforehand a review of the state of the art in this domain. Aiming at this objective, in this review we consider the most interesting publications since 2000. In essence, both a rapid approach with mainly immunoassays and chromatographic methods were developed. To date, the obstacle to routine use of the first approach has been its inability to detect more than two compounds simultaneously, but recent developments in flow cytometry have made it possible to detect six Coccidiostats at once. On the other hand, an increasingly popular approach for detecting multiple Coccidiostats simultaneously is liquid chromatography coupled with tandem mass spectrometry. There remains a need to adapt these analytical methods to legislative requirements.
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Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six Coccidiostats in feed and eggs
Analytical and Bioanalytical Chemistry, 2012Co-Authors: Monique E. Bienenmann-ploum, Terence L. Fodey, Philippe Delahaut, Ursula Vincent, Anne-catherine Huet, Katrina Campbell, Willem Haasnoot, Christopher T. Elliott, Michel W. F. NielenAbstract:Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these Coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority Coccidiostats. Different Coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation ( r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4′-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 μg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 μg/kg, respectively. Figure
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Development of a five-plex flow cytometric immunoassay for the simultaneous detection of six Coccidiostats in feed and eggs
Analytical and Bioanalytical Chemistry, 2012Co-Authors: Monique E. Bienenmann-ploum, Terence L. Fodey, Philippe Delahaut, Ursula Vincent, Anne-catherine Huet, Katrina Campbell, Willem Haasnoot, Christopher T. Elliott, Michel W. F. NielenAbstract:Coccidiostats are the only veterinary drugs still permitted to be used as feed additives to treat poultry for coccidiosis. To protect consumers, maximum levels for their presence in food and feed have been set by the European Union (EU). To monitor these Coccidiostats, a rapid and inexpensive screening method would be a useful tool. The development of such a screening method, using a flow cytometry-based immunoassay, is described. The assay uses five sets of colour-coded paramagnetic microspheres for the detection of six selected priority Coccidiostats. Different Coccidiostats, with and without carrier proteins, were covalently coupled onto different bead sets and tested in combination with polyclonal antisera and with a fluorescent-labelled secondary antibody. The five optimal combinations were selected for this multiplex and a simple-to-use sample extraction method was applied for screening blank and spiked eggs and feed samples. A very good correlation (r ranging from 0.995 to 0.999) was obtained with the responses obtained in two different flow cytometers (Luminex 100 and FLEXMAP 3D). The sensitivities obtained were in accordance with the levels set by the EU as the measured limits of detection for narasin/salinomycin, lasalocid, diclazuril, nicarbazin (4,4′-dinitrocarbanilide) and monensin in eggs were 0.01, 0.1, 0.5, 53 and 0.1 μg/kg and in feed 0.1, 0.2, 0.3, 9 and 1.5 μg/kg, respectively.
Dario Compagnone - One of the best experts on this subject based on the ideXlab platform.
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investigation of nonionophoric Coccidiostat residues in feed as a consequence of carryover
Journal of Food Protection, 2018Co-Authors: Loredana Annunziata, Pierina Visciano, Arianna Stramenga, Maria Novella Colagrande, Guido Campana, Giampiero Scortichini, Giacomo Migliorati, Dario CompagnoneAbstract:ABSTRACT Residues of nonionophoric Coccidiostats at carryover concentrations in feedstuffs collected from feed mills or animal farms in central Italy were detected as part of the official controls carried out from 2011 through 2016. The 118 samples were collected on the production line or during feed distribution and storage to determine the sampling sites at major risk of cross-contamination. For determination of nonionophoric Coccidiostats, a fast, easy, and cheap method was developed and validated. Feed samples were extracted with acetonitrile-methanol and directly injected for liquid chromatography with tandem mass spectrometry. A total of 24 samples (20.3%) were positive, but only 5 (4.2%) of these samples exceeded the maximum limit set by European legislation. Most of the positive samples were collected from a batch of feed produced immediately following processing of another batch to which the Coccidiostat robenidine had been added.
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determination of regulatory ionophore Coccidiostat residues in feedstuffs at carry over levels by liquid chromatography mass spectrometry
PLOS ONE, 2017Co-Authors: Loredana Annunziata, Pierina Visciano, Arianna Stramenga, Maria Novella Colagrande, Guido Campana, Giampiero Scortichini, Giacomo Migliorati, Dario CompagnoneAbstract:: In this study samples of feedstuffs were collected from different feed mills and animal farms located in central Italy and analyzed for ionophore Coccidiostat residues at carry-over levels by liquid chromatography-mass spectrometry. Since unavoidable cross-contamination of feedstuffs may occur during their production as well as distribution and storage, the collection of samples covered all these different stages. Residues of lasalocid, monensin, salinomycin and maduramicin were detected in 32.4% of samples, both at production and storage level. The maximum content for unavoidable carry-over set by Regulation (EU) No 574/2011 was exceeded in 11.3% of samples. The variability of the results highlighted the different approach of each investigated feed business operator to avoid any cross-contamination in non-target feed. The method developed in this study can be able to detect ionophore Coccidiostats at low concentrations consequent to carry-over.