This standard describes a method of Iodium-131, Caesium-134 and Caesium-137 massic activity (Bq/kg) determination in animal feed.
Today, the most commonly used method for identification and quantification of radioactivity from these radionuclides in feed samples is high-resolution gamma-ray spectrometry. It is based on analysis of full-energy peaks (FEP) of the emitted gamma rays. Therefore, care should be taken to use appropriate energy and efficiency calibrations for each detector and test portion used.
In this standard, general guidance on the preparation of feed samples is provided together with specific information on high resolution gamma-ray spectrometry of the three radionuclides Iodium-131, Caesium-134 and Caesium-137. More information on these and related topics can be found in specific standards referred to in this document. For example, generic advice on the equipment selection, detectors and quality assurance for gamma-ray spectrometry can be found in ISO 20042:2016. The current standard aims to be complementary to existing standards, as an aid to laboratory practitioners that are faced with a situation, which requires response to the current topic without having to go through and interpret standards with general descriptions of gamma-ray spectrometry in order to measure in a standardised way. This standard contains information specific to the three radionuclides that it covers. Examples are provided in Annex …(to be added).

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This method procedure describes a procedure for the determination of inorganic arsenic in animal feeding stuffs by anion-exchange HPLC-ICP-MS following water bath extraction.

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This document specifies a liquid chromatographic method with triple-quadrupole mass spectrometry (MS/MS) detection for the determination of pentachlorophenol (PCP) in feed materials and animal feed.
The limit of quantitation (LOQ) for the PCP determination in guar gum, fatty acid distillates (FAD) and animal feed is 10 µg/kg. Individual laboratories are responsible for ensuring that the equipment that they use will achieve this limit of quantification.
The method is validated in an international collaborative trial for pentachlorophenol in compound feed, guar gum and fatty acid distillate in the range between 9 µg/kg and 22 µg/kg.
The results of the collaborative trial, in which 16 laboratories participated, have shown that the method is applicable for the determination of PCP in compound feed, guar gum and FAD at the desired limit of 10 µg/kg. Satisfactory results were obtained for one compound feed sample, guar gum and the two FAD samples (HorRat <2), while for the second compound feed sample a HorRat value of 2,2 was obtained.

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This document gives guidance to those involved in designing, executing and evaluating interlaboratory comparison studies for multi-analyte methods of analysis, developed by CEN/TC 327 “Animal feeding stuffs: Methods of sampling and analysis” and its working groups.
For the validation of multi-analyte methods their particularities must be considered which might necessitate deviations from the prescribed validation protocols. This study provides information whether the method is fit for its purpose and which performance can be expected in practical work while at the same time keeping the necessary effort for the study organizer and the participating laboratories minimal.
Next to the abovementioned aspects regarding interlaboratory comparison studies, this document also gives guidance on the preceding steps, viz. in-house validation and preparation of the method protocol. Guidance is also given on the transferability of the method protocol and the familiarization with the method protocol through a training study, elements that – depending on the specific method – could be included in the design of the study.

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  • Technical report
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This document gives guidance to those involved in designing, executing and evaluating interlaboratory comparison studies for multi-analyte methods of analysis, developed by CEN/TC 327 “Animal feeding stuffs: Methods of sampling and analysis” and its working groups.
For the validation of multi-analyte methods their particularities must be considered which might necessitate deviations from the prescribed validation protocols. This study provides information whether the method is fit for its purpose and which performance can be expected in practical work while at the same time keeping the necessary effort for the study organizer and the participating laboratories minimal.
Next to the abovementioned aspects regarding interlaboratory comparison studies, this document also gives guidance on the preceding steps, viz. in-house validation and preparation of the method protocol. Guidance is also given on the transferability of the method protocol and the familiarization with the method protocol through a training study, elements that – depending on the specific method – could be included in the design of the study.

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  • Technical report
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This document specifies a liquid chromatographic method with triple-quadrupole mass spectrometry (MS/MS) detection for the determination of pentachlorophenol (PCP) in feed materials and animal feed.
The limit of quantitation (LOQ) for the PCP determination in guar gum, fatty acid distillates (FAD) and animal feed is 10 µg/kg. Individual laboratories are responsible for ensuring that the equipment that they use will achieve this limit of quantification.
The method is validated in an international collaborative trial for pentachlorophenol in compound feed, guar gum and fatty acid distillate in the range between 9 µg/kg and 22 µg/kg.
The results of the collaborative trial, in which 16 laboratories participated, have shown that the method is applicable for the determination of PCP in compound feed, guar gum and FAD at the desired limit of 10 µg/kg. Satisfactory results were obtained for one compound feed sample, guar gum and the two FAD samples (HorRat <2), while for the second compound feed sample a HorRat value of 2,2 was obtained.

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This document describes a method for determination of the massic activity (Bq/kg) of 131I, 134Cs and 137Cs in animal feeding stuffs in monitoring laboratories.
General guidance on the preparation of feed samples and the measurement of the three radionuclides 131I, 134Cs and 137Cs by high resolution gamma-ray spectrometry is provided. The current document aims to be complementary to existing standards. More information on sample preparation, moisture content determination and gamma-ray spectrometry can be found in specific standards referred to in this document. For example, generic advice on the equipment selection, detectors and quality assurance for gamma-ray spectrometry can be found in ISO 20042 [4].
The method was fully statistically tested and evaluated in a collaborative trial comprising five animal feeding stuff samples for the radionuclides 131I, 134Cs and 137Cs. Details on the successfully tested working range for each of the examined radionuclides are described in Annex C.

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This document specifies a procedure for the determination of inorganic arsenic in animal feeding stuffs by anion-exchange HPLC-ICP-MS following water bath extraction.
This method was successfully tested in the range of 0,149 mg/kg to 9,69 mg/kg in the following animal feed matrices: rice meal, seaweed meal, fish meal, grass meal, complete feed (marine-based), complete feed (cereal based) and a synthetic solution.
NOTE   Mineral feed matrices are not included in the scope of this method. It is good to perform a determination of the total arsenic content in such matrices.

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This analytical procedure specifies a reverse phase high performance liquid chromatographic with UV detection (RP-HPLC-UV) method for the simultaneous determination of four authorized carotenoids in fish compound feed, namely astaxanthin (AXN), canthaxanthin (CXN), adonirubin (ADR) and astaxanthin dimethyldisuccinate (AXN DMDS), and of six authorized carotenoids in poultry feed, namely canthaxanthin (CXN); capsanthin (CSN), ethyl ester of beta-apo-8'-carotenoic acid (BACARE), citranaxanthin (CIXN), lutein (LUT) and zeaxanthin (ZEA) at levels ranging from ca. 2 to ca. 4 500 mg/kg (depending on the carotenoid). Beta-carotene (BCAR), authorized in compound feed for all animal species, was also added to the scope. The analytical procedure is fit for the purpose of quantitation of declared carotenoids and labelling confirmation. The procedure applies to natural and synthetic feed additives.
Xanthophyll esters like those of lutein, zeaxanthin and capsanthin that might be present in feed materials are not authorized feed additives and therefore not part of the scope of this method.

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This European Standard specifies a method for the determination of the content of the total vitamin A (retinol), vitamin E (alpha-tocopherol) and vitamin D (D2 ergocalciferol or D3 cholecalciferol) content in animal feed using solid phase extraction (SPE) clean-up and high performance liquid chromatography (HPLC).
The limit of quantification is XXXX IU vitamin A/kg (using UV-detection), XX IU vitamin A/kg (using fluorescence detection), XX mg vitamin E/kg (using UV-detection), XX mg vitamin E/kg (using
fluorescence detection), XX IU vitamin D/kg (using UV-detection) and XX IU vitamin D/kg (using fluorescence detection).

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This document specifies a method for the determination of saturated and aromatic hydrocarbons (from C10 to C50) in feed. The method has been interlaboratory validated with online-HPLC-GC-FID – see [1], [2] and [3]. This method is not intended to be applied to other matrices.
The method can be used for the analysis of mineral oil saturated hydrocarbons (MOSH) and/or mineral oil aromatic hydrocarbons (MOAH).
The method is applicable for feed materials, in particular vegetable oils and other fat rich feed materials, compound feeds and pre-mixtures. It is not applicable to additives or deodistillates.
The method has been tested in an interlaboratory study via the analysis of both naturally contaminated and spiked samples (pre-mixture, soybean meal, sunflower seeds, chicken feed, pig feed, vegetable oil) ranging from 3 mg/kg to 286 mg/kg for MOSH and from 1 mg/kg to 16 mg/kg for MOAH.
According to the results of the interlaboratory study, the method has been proven suitable for MOSH and MOAH mass concentrations, each above 10 mg/kg. However, the method was not fully validated during the collaborative study for the premixture sample due to too low concentrations of MOSH and MOAH. The method was also not fully validated during the collaborative study for the sunflower seeds sample due to a too low concentration of MOAH.
NOTE   The conclusions regarding MOAH are based on 4 analyte / matrix combinations while according to the IUPAC protocol [4] expects this to be a minimum of 5.
In case of suspected interferences from natural sources, the fossil origin of the MOSH and MOAH fraction can be verified by examination of the pattern by GC-MS.
For the determination of MOSH and MOAH in edible fats and oils, another CEN standard is also available: EN 16995. For more information see [5].
Annex C proposes a manual alternative method to online HPLC-GC-FID analysis that can be used as a screening method.

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This document describes a method for the determination of free gossypol, extractable by acidified
acetonitrile/water, in cottonseeds, cottonseed cake and complete feed by high performance liquid
chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS)
This method has been in-house validated in the range 20-6000 mg/kg.

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This analytical procedure specifies a reverse phase high performance liquid chromatographic with UV detection (RP-HPLC-UV) method for the simultaneous determination of four authorized carotenoids in fish compound feed and fish premix, namely astaxanthin (AXN), canthaxanthin (CXN), adonirubin (ADR) and astaxanthin dimethyldisuccinate (AXN DMDS), and of six authorized carotenoids in poultry feed and poultry premix, namely canthaxanthin (CXN); capsanthin (CSN), ethyl ester of beta-apo-8'-carotenoic acid (BACARE), citranaxanthin (CIXN), lutein (LUT) and zeaxanthin (ZEA) at levels ranging from approximately 2 mg/kg to approximately 4 500 mg/kg (depending on the carotenoid). Beta-carotene (BCAR), authorized in compound feed and premixes for all animal species, was also added to the scope. The analytical procedure is fit for the purpose of quantitation of declared carotenoids and labelling confirmation. This document is applicable to feed produced using natural and synthetic feed additives.
Xanthophyll esters like those of lutein, zeaxanthin and capsanthin that might be present in feed materials are not authorized feed additives and therefore not part of the scope of this document.

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This document describes a method for the determination of free gossypol, extractable by acidified
acetonitrile/water, in cottonseeds, cottonseed cake and complete feed by high performance liquid
chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS)
This method has been in-house validated in the range 20-6000 mg/kg.

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This European Standard specifies a method for the determination of the content of the total vitamin A (retinol), vitamin E (alpha-tocopherol) and vitamin D (D2 ergocalciferol or D3 cholecalciferol) content in animal feed using solid phase extraction (SPE) clean-up and high performance liquid chromatography (HPLC).
The limit of quantification is XXXX IU vitamin A/kg (using UV-detection), XX IU vitamin A/kg (using fluorescence detection), XX mg vitamin E/kg (using UV-detection), XX mg vitamin E/kg (using
fluorescence detection), XX IU vitamin D/kg (using UV-detection) and XX IU vitamin D/kg (using fluorescence detection).

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This document specifies a method for the determination of saturated and aromatic hydrocarbons (from C10 to C50) in feed. The method has been interlaboratory validated with online-HPLC-GC-FID - see [1], [2] and [3]. This method is not intended to be applied to other matrices.
The method can be used for the analysis of mineral oil saturated hydrocarbons (MOSH) and/or mineral oil aromatic hydrocarbons (MOAH).
The method is applicable for feed materials, in particular vegetable oils and other fat rich feed materials, compound feeds and pre-mixtures. It is not applicable to additives or deodistillates.
NOTE 1    The method was not designed for encapsulated matrices.
The method has been tested in an interlaboratory study via the analysis of both naturally contaminated and spiked samples (pre-mixture, soybean meal, sunflower seeds, chicken feed, pig feed, vegetable oil) ranging from 3 mg/kg to 286 mg/kg for MOSH and from 1 mg/kg to 16 mg/kg for MOAH.
According to the results of the interlaboratory study, the method has been proven suitable for MOSH and MOAH mass concentrations, each above 10 mg/kg. However, the method was not fully validated during the collaborative study for the premixture sample due to too low concentrations of MOSH and MOAH. The method was also not fully validated during the collaborative study for the sunflower seeds sample due to a too low concentration of MOAH.
NOTE 2   The conclusions regarding MOAH are based on 4 analyte / matrix combinations while according to the IUPAC protocol [4] expects this to be a minimum of 5.
In case of suspected interferences from natural sources, the fossil origin of the MOSH and MOAH fraction can be verified by examination of the pattern by GC-MS.
For the determination of MOSH and MOAH in edible fats and oils, another CEN standard is also available: EN 16995. For more information see [5].
Annex C proposes a manual alternative method to online HPLC-GC-FID analysis that can be used as a screening method for the determination of MOSH.

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