EURL-FA Control Proficiency test report: Determination of authorised coccidiostats in compound feed

The European Union Reference Laboratory for Feed Additives (EURL-FA), hosted by the Joint Research Centre (JRC), a Directorate General of the European Commission, has been mandated by the Directorate General for Health and Food Safety (DG SANTE) to organise a proficiency test (PT) among appointed National Reference Laboratories (NRLs) in the frame of its control activities (according to the Regulation (EC) No 882/2004 [1]). The aim of this PT was to assess the capacity of the NRLs to correctly determine selected authorised coccidiostats added to feed matrices at realistic authorised levels and at cross-contamination levels. Thirty-six European National and Official Control laboratories were invited and twenty-nine laboratories registered to the 2017 PT exercise. Twenty-five of the registered laboratories reported results for the analyses. The test items used in this exercise were produced by the JRC. Purchased commercial poultry compound feed, tested by the EURL-FA Control as being blank for the target analytes, was milled and ground and then spiked with the required coccidiostat standard solution or with the relevant authorised feed additive. The first item was spiked with a standard solution containing monensin, narasin and diclazuril, at cross-contamination level (MAT 1). The second test item (MAT 2) was spiked with an authorised feed additive Maxiban®, containing narasin (narasin AL) and nicarbazin, at additive level. MAT 1 and MAT 2 were subsequently homogenised and distributed in glass bottles. All bottles were labelled ensuring a random number encoding and dispatched to all registered participants on 27 June 2017. Laboratories were informed of the composition of the test material regarding the composition in coccidiostats for MAT 2 and had therefore only to quantify the content. For MAT 1 the laboratories had to screen for the presence of all 11 authorised coccidiostats and to quantify the detected ones. The assigned values (xpt) for the mass content of monensin, diclazuril and narasin in MAT 1 were calculated from the formulation as recommended by the IUPAC harmonized protocol [2]. The uncertainties for the assigned values (u(xpt)) were calculated according to the ISO Guide for the Expression of Uncertainty in Measurement (GUM) [3]. For narasin and nicarbazin in MAT 2, no assigned value was set since the criterion for sufficient homogeneity could not be met. Participants were invited to report their measurement uncertainties. This was done by twenty-three out of twenty-five reporting participants for monensin and narasin, seventeen for diclazuril, and twenty for narasin AL and nicarbazin. Laboratory results were rated using z and ζ (zeta) scores in accordance with ISO 13528:2015 [4]. The relative standard deviation for proficiency assessment (σpt) for each assigned value was calculated using the relevant Horwitz [5] or modified Horwitz equation [6]. The z scores obtained were considered satisfactory if their absolute values were equal to or below 2. The outcome of this PT exercise is mixed; the percentage of satisfactory results reported by the laboratories for MAT 1 is 88%, 80% and 57% for monensin, narasin and diclazuril respectively. For narasin AL and nicarbazin in MAT 2, no scoring was computed but the data reported by the laboratories were examined. In general, there was good agreement among the values of mass content of narasin and nicarbazin reported results. The laboratories also reported qualitative results as regards the presence of one or more of the other authorised coccidiostats. On the whole, the rate of false positive results was of 4% for robenidine, lasalocid, salinomycin and maduramicin; 5% for nicarbazin; 8% for decoquinate; and 0% for all the others. Two laboratories did not quantify diclazuril in MAT 1 while stating a limit of quantification of the method used lower than the assigned value, leading to a false negative rate of 9% for this analyte. One laboratory could not quantify diclazuril in MAT 1 due to the lack of sensitivity of the method used.JRC.F.5-Food and Feed Complianc

EURL-FA Control Proficiency Test Report: Determination of the mass fraction of the total cobalt in compound feed for rabbits

The European Union Reference Laboratory for Feed Additive Control (EURL-FA Control) organised a proficiency test (EURL-FAC 2017-2) for the determination of the mass fraction of total cobalt in compound feed, to support the Commission Implementing Regulation (EU) No 131/2014 concerning the authorisation of various cobalt(II) compounds as feed additives. This proficiency test was open to National Reference Laboratories (NRLs) and official feed control laboratories (OCLs). The material used as test item was a commercially available compound feed for rabbits (containing the coated granulated cobalt (II) carbonate, 3b304 ) which, after appropriate processing, was bottled, labelled and dispatched to participants on June 21, 2017. The homogeneity and stability of the test item were evaluated and the assigned values were derived from the results reported by the selected expert laboratories. Twenty one NRLs and two OCLs from 21 countries - representing EU Member States and Norway - registered to the exercise and reported results at the end of August 2017. Laboratory results were rated using z' and zeta scores in accordance with ISO 13528:2015. A relative standard deviation for proficiency assessment (σpt) of 16 % of the assigned value was set according to the modified Horwitz equation. Twenty (out of 23) laboratories reported satisfactory results (according to the z' score). This confirms the ability of most NRLs in monitoring maximum levels set by the Commission Implementing Regulation (EU) No 131/2014 in this type of animal feed. The majority (74 %) of the participating laboratories provided realistic estimates of their measurement uncertainties.JRC.F.5-Food and Feed Complianc

Proficiency of official control European laboratories in the determination of authorised coccidiostats in poultry feed - EURL Feed Additives Control, PT exercise 2014, Cross-contamination and additive levels

The European Union Reference Laboratory for Feed additives, has been mandated by the Directorate General for Health and Consumers to organise a proficiency test (PT) among appointed National Reference Laboratories (NRLs). The aim of the PT was to assess the capacity of the NRLs to correctly determine 11 authorised coccidiostats in feed matrices at cross-contamination and additive levels. This report presents the results of the PT. Thirty-eight laboratories from 22 countries registered to this 2014 PT exercise. Laboratory results were rated using z- and ζ-scores (zeta-scores). Between 63 % and 79 % of the laboratories reported satisfactory results for monensin, narasin, lasalocid, diclazuril, decoquinate, halofuginone, robenidine, semduramicin and diclazuril FA. The laboratories also reported qualitative results as regards the presence of one or more of the other authorised coccidiostats. On the whole, the rate of false positive results was 3% for monensin and maduramicin and 0% for all the others.JRC.D.5-Standards for Food Bioscienc

Determination of authorised coccidiostats in poultry compound feed: EURL Feed Additives Control, Proficiency test exercise 2012

The European Union Reference Laboratory for Feed Additives Control (EURL-FA Control) has been mandated by the Directorate General for Health and Consumers (DG SANCO) to organise a Proficiency Test exercise (PT) among appointed National Reference Laboratories (NRLs) in order to assess their capacity to correctly determine the 11 authorised coccidiostats in feed matrices. This report presents the results of the PT. Thirty laboratories from 22 countries registered to the exercise. Laboratory results were rated using z- and ζ-scores (zeta-scores). Between 62% and 73 % of the laboratories reported satisfactory results for four of the five spiked measurands, namely monensin, lasalocid, diclazuril and salinomycin. For the fifth spiked one, narasin, only 57 % of the laboratories submitted satisfactory results. The laboratories also reported qualitative results as regards to the presence of one or more of the other authorised coccidiostats. On the whole, the rate of false positive results was 3% for monensin and narasin, 4% for robenidine, 7% for lasalocid and 0% for all the others.JRC.D.5-Standards for Food Bioscienc

Proficiency of European National Reference Laboratories in the determination of authorised vitamins in animal feed. EURL for Feed Additives Control PT exercise 2016 and annual workshop.

The European Union Reference Laboratory for Feed Additives (EURL-FA), hosted by the Joint Research Centre (JRC), a Directorate General of the European Commission, has been mandated by the Directorate General for Health and Food Safety (DG SANTE) to organise a proficiency test (PT) among appointed National Reference Laboratories (NRLs) in the frame of its control activities (according to the Regulation (EC) No 882/2004 [1]). The aim of this PT was to assess the capacity of the NRLs to correctly determine selected authorised vitamins added to feed matrices at realistic levels. Twenty-eight European National and Official Control laboratories were invited and eleven laboratories registered to the 2016 PT exercise. The eleven registered laboratories reported results for the analyses. The test items used in this exercise were produced by the EURL-FA Control by milling purchased commercial poultry compound feed containing vitamin A and vitamin E, homogenising and assessing the mass content versus the declared mass content on the label. Each laboratory received one test item. Laboratories were informed of the composition of the test material regarding vitamins and the label provided information on the mass content range to be expected for each vitamin as well as the additive code as specified in the European Union register of feed additives [2]. The assigned values (xa) for the mass content of vitamin A and vitamin E in the samples were established by calculating the robust mean of the measurements performed by the EURL-FA Control and the sub-group of participants who applied the protocol specified in Annex IV of Commission Regulation (EC) No 152/2009 [3] and that were accredited according to ISO 17025:2005 [4] for this protocol. For the uncertainties of the assigned values (Ua), the associated expanded uncertainty (coverage factor k=2) was derived from the robust standard error on the measurements performed by the EURL-FA Control and the sub-group defined above and the between-sample standard deviation of the results obtained during the homogeneity study, calculated using ANOVA. Participants were invited to report the uncertainties of their measurements for completeness of result reporting. This was done by ten out of eleven participants. Laboratory results were rated using z-scores in accordance with ISO 13528:2015 [5]. The standard deviation for proficiency assessment (σp) for each assigned value was set beforehand applying the modified Horwitz equation [6] to the content determined during the homogeneity study performed at the EURL-FA Control. The z-scores obtained were considered satisfactory if their absolute values were equal to or below 2. The outcome of the PT exercise is satisfactory; the percentage of satisfactory results reported by the laboratories is 91% for vitamin A and 91% for vitamin E, respectively.JRC.F.5-Food and Feed Complianc

Determination of (fluoro)Quinolone Antibiotic Residues in Pig Kidney Using Liquid Chromatography-tandem Mass Spectrometry Part II: Intercomparison Exercise

A recently in-house validated method for the liquid chromatography-tandem mass spectrometry (LC-MS/MS) determination of eleven (fluoro)quinolone antibiotics (FQs) in pig kidney has been fully validated through an intercomparison exercise. This ring trial involved eight European Laboratories and was based on the Commission Decision 2002/657/CE for validation of method and on the IUPAC protocol for method performances studies. The laboratories data were submitted to a one-way analysis of variance. Satisfactory results were obtained for each FQ with regards to within- and between-laboratory reproducibility and accuracy. The method was validated for the simultaneous qualitative and quantitative determination of the eleven FQs in pig kidney around their maximum residue limit (MRL) as defined in the European Council Regulation 2377/90/EEC.JRC.D.8-Food safety and qualit

Determination of (fluoro)Quinolone Antibiotic Residues in Pig Kidney using Liquid Chromatography-tandem Mass Spectrometry. I. Laboratory-validated Method

A new LC-MS/MS method has been developed for the multiresidue determination of 11 (fluoro)quinolone antibiotics (FQs), including acidic and aphoteric speices, around their maximum residue level (MRL) in pig kidney. The procedure involved a common sample preparation by solid-phase extraction on disposable extraction cartridges followed by a fast reversed-phase liquid chromatography-tandem mass spectrometry analysis. The method was validated according to the Commission Decision 2002/657/CE. The accuracy of the method was satisfactory with recoveries included in the interval 80-100%. The precision results showed mean repeatability and reproducibility coefficients of 7.4% and 11.8% respectively. Limits of quantification much lower than the MRLs could be obtained.JRC.D.8-Food safety and qualit

Proficiency of official control European laboratories in the determination of authorised coccidiostats in poultry feed - EURL Feed Additives Control, PT exercise 2013, Cross-contamination level

The European Union Reference Laboratory for Feed additives (control) (EURL-FA (control)) has been mandated by the Directorate General for Health and Consumers (DG SANCO) to organise an interlaboratory comparison (ILC) among appointed National Reference Laboratories (NRLs) in order to assess their capacity to correctly determine the 11 authorised coccidiostats at cross-contamination levels in feed matrices. Other EU control laboratories and one laboratory from Switzerland also joined the exercise. This report presents the results of the proficiency test (PT). Thirty-two laboratories from 22 countries registered to the exercise. Three test items were produced, homogenised and distributed by the EURL-FA Control. Two contained 4 and 3 selected coccidiostats respectively and the third one was a blank. Laboratory results were rated using z- and ζ-scores (zeta-scores). Between 64% and 80 % of the laboratories reported satisfactory results for monensin, narasin, nicarbazin and maduramicin. For lasalocid, only 59 % of the laboratories submitted satisfactory results, 58% for diclazuril and 53% for halofuginone. The results are summarised in the following Table 1. The laboratories also reported qualitative results as regards the presence of one or more of the other authorised coccidiostats. On the whole, the rate of false positive results was 3% for lasalocid, 4% for maduramicin, 5% for halofuginone and 0% for all the others.JRC.D.5-Standards for Food Bioscienc

Determination of Ionophore Coccidiostats in Feedingstuffs by Liquid Chromatography-Tandem Mass Spectrometry Part I. Application to Targeted Feed

A new and fit to purpose multi-analyte method for the determination of six coccidiostats (monensin A, salinomycin, narasin, composed of its principle components 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. 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 87 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 mg kg and 0.046 mg kg 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 complimentary to the official PHLC-UV methods.JRC.D.8-Food safety and qualit

Determination of (fluoro)quinolone Antibiotic Resideus in Pig kidney Using LC-MS/MS. Part II: Interncomparison Excercise

Abstract not availableJRC.D-Institute for Reference Materials and Measurements (Geel