882/2004 - Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 onofficial controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules

This document specifies a method for the identification of single fish and fish fillets to the level of genus or species. It allows the identification of a large number of commercially important fish species using DNA barcoding.
This method was validated on raw fish. Laboratory experience indicates additional applicability to processed fish products (e.g. cold smoked, hot smoked, salted, frozen, cooked, fried and deep-fried samples).
The described method is usually unsuitable for the analysis of highly processed foods (e.g. tins of fish with highly degraded DNA where the fragment lengths are not sufficient for amplification of the targets). Furthermore, it does not apply to complex fish products containing mixtures of two or more fish species.
The identification of fish species is carried out by PCR amplification of either a segment of the mitochondrial cytochrome b gene (cytb) or the cytochrome c oxidase I gene (cox1, syn COI), or both, followed by sequencing of the PCR products and subsequent sequence comparison with entries in databases.

This document specifies a method for the identification of single fish and fish fillets to the level of genus or species. It allows the identification of a large number of commercially important fish species using DNA barcoding.
This method was validated on raw fish. Laboratory experience indicates additional applicability to processed fish products (e.g. cold smoked, hot smoked, salted, frozen, cooked, fried and deep-fried samples).
The described method is usually unsuitable for the analysis of highly processed foods (e.g. tins of fish with highly degraded DNA where the fragment lengths are not sufficient for amplification of the targets). Furthermore, it does not apply to complex fish products containing mixtures of two or more fish species.
The identification of fish species is carried out by PCR amplification of either a segment of the mitochondrial cytochrome b gene (cytb) or the cytochrome c oxidase I gene (cox1, syn COI), or both, followed by sequencing of the PCR products and subsequent sequence comparison with entries in databases.

This document specifies a method for the taxonomic identification of a single bivalve or piece of bivalve to the genus or species level using DNA barcoding. It allows the identification of a large number of commercially important bivalve species.
This method was validated on raw mussels. Laboratory experience indicates additional applicability to processed bivalve products, e.g. cold smoked, hot smoked, salted, frozen, cooked, fried, and deep-fried samples.
The described method is usually unsuitable for the analysis of highly processed foods, e.g. tins of mussels, with highly degraded DNA where the fragment lengths are not sufficient for amplification of the target. Furthermore, it is not applicable for complex seafood products containing mixtures of two or more bivalve species.
The identification of bivalve species is carried out by PCR amplification of a segment of the mitochondrial 16S rRNA gene, followed by sequencing of the PCR products and subsequent sequence comparison with entries in databases.

This document specifies a method for the identification of meat derived from mammals and birds to the level of genus or species and allows the identification of a large number of commercially important as well as exotic meat species using DNA barcoding.
This method was validated on DNA isolated from single pieces of raw meat. This method can also be used for the identification of single meat animal species in some processed products.
The described method is unsuitable for the analysis of highly processed foods with highly degraded DNA where the fragment lengths are not sufficient for amplification of the targets. Furthermore, it is not applicable for complex meat products containing mixtures of two or more meat species.
The identification of meat species is carried out by PCR amplification of either a segment of the mitochondrial cytochrome b gene (cytb) or the cytochrome c oxidase I gene (cox1, syn COI) or both, followed by sequencing of the PCR products and subsequent sequence comparison with entries in databases.

This document specifies a method for the identification of meat derived from mammals and birds to the level of genus or species and allows the identification of a large number of commercially important as well as exotic meat species using DNA barcoding.
This method was validated on DNA isolated from single pieces of raw meat. This method can also be used for the identification of single meat animal species in some processed products.
The described method is unsuitable for the analysis of highly processed foods with highly degraded DNA where the fragment lengths are not sufficient for amplification of the targets. Furthermore, it is not applicable for complex meat products containing mixtures of two or more meat species.
The identification of meat species is carried out by PCR amplification of either a segment of the mitochondrial cytochrome b gene (cytb) or the cytochrome c oxidase I gene (cox1, syn COI) or both, followed by sequencing of the PCR products and subsequent sequence comparison with entries in databases.

This document specifies a method for the taxonomic identification of a single bivalve or piece of bivalve to the genus or species level using DNA barcoding. It allows the identification of a large number of commercially important bivalve species.
This method was validated on raw mussels. Laboratory experience indicates additional applicability to processed bivalve products, e.g. cold smoked, hot smoked, salted, frozen, cooked, fried, and deep-fried samples.
The described method is usually unsuitable for the analysis of highly processed foods, e.g. tins of mussels, with highly degraded DNA where the fragment lengths are not sufficient for amplification of the target. Furthermore, it is not applicable for complex seafood products containing mixtures of two or more bivalve species.
The identification of bivalve species is carried out by PCR amplification of a segment of the mitochondrial 16S rRNA gene, followed by sequencing of the PCR products and subsequent sequence comparison with entries in databases.

This document specifies the detection of Clostridium (C.) perfringens.
This document is applicable to:
—     products intended for human consumption;
—     products intended for animal feeding;
—     environmental samples in the area of food and feed production and handling;
—     samples from the primary production stage.
This horizontal method was originally developed for the examination of all samples belonging to the food chain. Based on the information available at the time of publication of this document, this method is considered to be fully suited to the examination of all samples belonging to the food chain. However, because of the large variety of products in the food chain, it is possible that this horizontal method is not appropriate in every detail for all products. Nevertheless, it is expected that the required modifications are minimized so that they do not result in a significant deviation from this horizontal method.
NOTE          Interlaboratory studies with a small number of participating laboratories (<10) were conducted for the following food categories:
—      ready-to-eat, ready-to-reheat meat products;
—      eggs and egg products (derivates);
—      ready-to-eat, ready-to-reheat fishery products;
—      processed fruits and vegetables;
—      infant formula and infant cereals (with probiotics);
—      multi-component foods or meal components.
It has also been validated with a small number of participating laboratories for the following other category:
—      environmental samples (food or feed production).
Since the method is not commonly used for samples in the primary production stage, this category was not included in the interlaboratory study. Therefore, no performance characteristics were obtained for this category. The method has not been validated for the category ‘pet food and animal feed’, as the test samples used for the interlaboratory study were already naturally contaminated with C. perfringens. Given the limited number of participating laboratories in the interlaboratory studies, the calculated performance characteristics can be used as indicative values of the method performance. For detailed information on the validation, see Clause 11 and Annexes C to F.

This document specifies the detection of Clostridium (C.) perfringens.
This document is applicable to:
—     products intended for human consumption;
—     products intended for animal feeding;
—     environmental samples in the area of food and feed production and handling;
—     samples from the primary production stage.
This horizontal method was originally developed for the examination of all samples belonging to the food chain. Based on the information available at the time of publication of this document, this method is considered to be fully suited to the examination of all samples belonging to the food chain. However, because of the large variety of products in the food chain, it is possible that this horizontal method is not appropriate in every detail for all products. Nevertheless, it is expected that the required modifications are minimized so that they do not result in a significant deviation from this horizontal method.
NOTE          Interlaboratory studies with a small number of participating laboratories (<10) were conducted for the following food categories:
—      ready-to-eat, ready-to-reheat meat products;
—      eggs and egg products (derivates);
—      ready-to-eat, ready-to-reheat fishery products;
—      processed fruits and vegetables;
—      infant formula and infant cereals (with probiotics);
—      multi-component foods or meal components.
It has also been validated with a small number of participating laboratories for the following other category:
—      environmental samples (food or feed production).
Since the method is not commonly used for samples in the primary production stage, this category was not included in the interlaboratory study. Therefore, no performance characteristics were obtained for this category. The method has not been validated for the category ‘pet food and animal feed’, as the test samples used for the interlaboratory study were already naturally contaminated with C. perfringens. Given the limited number of participating laboratories in the interlaboratory studies, the calculated performance characteristics can be used as indicative values of the method performance. For detailed information on the validation, see Clause 11 and Annexes C to F.

This document specifies the enumeration of Clostridium (C.) perfringens by colony-count technique.
This document is applicable to:
—    products intended for human consumption;
—    products for feeding animals;
—    environmental samples in the area of food and feed production and handling;
—    samples from the primary production stage.
NOTE            This method has been validated in an interlaboratory study for the following food categories:
—     ready-to-eat, ready-to-reheat meat products;
—     eggs and egg products (derivates);
—     processed fruits and vegetables;
—     infant formula and infant cereals;
—     multi-component foods or meal components.
It has also been validated for the following other categories:
—     pet food and animal feed;
—     environmental samples (food or feed production).
As this method has been validated for at least five food categories, this method is applicable for a broad range of food. For detailed information on the validation, see Clause 11 and Annex C. Since the method is not commonly used for samples in the primary production stage, this category was not included in the interlaboratory study. Therefore, no performance characteristics were obtained for this category.
This horizontal method was originally developed for the examination of all samples belonging to the food chain. Based on the information available at the time of publication of this document, this method is considered to be fully suited to the examination of all samples belonging to the food chain. However, because of the large variety of products in the food chain, it is possible that this horizontal method is not appropriate in every detail for all products. Nevertheless, it is expected that the required modifications are minimized so that they do not result in a significant deviation from this horizontal method.
This technique is suitable for, but not limited to, the enumeration of microorganisms in test samples with a minimum of 10 colonies counted on a plate. This corresponds to a level of contamination that is expected to be higher than 10 cfu/ml for liquid samples or higher than 100 cfu/g for solid samples.

This document specifies the enumeration of Clostridium (C.) perfringens by colony-count technique.
This document is applicable to:
—    products intended for human consumption;
—    products for feeding animals;
—    environmental samples in the area of food and feed production and handling;
—    samples from the primary production stage.
NOTE            This method has been validated in an interlaboratory study for the following food categories:
—     ready-to-eat, ready-to-reheat meat products;
—     eggs and egg products (derivates);
—     processed fruits and vegetables;
—     infant formula and infant cereals;
—     multi-component foods or meal components.
It has also been validated for the following other categories:
—     pet food and animal feed;
—     environmental samples (food or feed production).
As this method has been validated for at least five food categories, this method is applicable for a broad range of food. For detailed information on the validation, see Clause 11 and Annex C. Since the method is not commonly used for samples in the primary production stage, this category was not included in the interlaboratory study. Therefore, no performance characteristics were obtained for this category.
This horizontal method was originally developed for the examination of all samples belonging to the food chain. Based on the information available at the time of publication of this document, this method is considered to be fully suited to the examination of all samples belonging to the food chain. However, because of the large variety of products in the food chain, it is possible that this horizontal method is not appropriate in every detail for all products. Nevertheless, it is expected that the required modifications are minimized so that they do not result in a significant deviation from this horizontal method.
This technique is suitable for, but not limited to, the enumeration of microorganisms in test samples with a minimum of 10 colonies counted on a plate. This corresponds to a level of contamination that is expected to be higher than 10 cfu/ml for liquid samples or higher than 100 cfu/g for solid samples.

This document specifies a method for the determination of individual intact glucosinolates in feed materials including oilseeds and oilseed products and in compound feeds by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS).
The method specified in this document has been successfully validated by collaborative trial in the following matrices: rape seed, camelina seed, Brassica oleracea seeds, mixed oilseeds, rape seed flakes, compound feed for bovine, porcine and poultry.
The method is applicable for the quantitative determination of epiprogroitrin, glucoalyssin, glucoarabin, glucobrassicanapin, glucobrassicin, glucocamelinin, glucoerucin, glucoiberin, gluconapin, gluconapoleiferin, gluconasturtiin, glucoraphanin, glucoraphenin, glucotropaeolin, homoglucocamelinin, 4-hydroxyglucobrassicin, 4-methoxyglucobrassicin, neoglucobrassicin, progoitrin, sinalbin and sinigrin.
The concentration ranges tested in the collaborative trial for each individual glucosinolate and for the total glucosinolate content are summarized in Table 1.
[table not represented]

This document specifies a method for the determination of individual intact glucosinolates in feed materials including oilseeds and oilseed products and in compound feeds by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS).
The method specified in this document has been successfully validated by collaborative trial in the following matrices: rape seed, camelina seed, Brassica oleracea seeds, mixed oilseeds, rape seed flakes, compound feed for bovine, porcine and poultry.
The method is applicable for the quantitative determination of epiprogroitrin, glucoalyssin, glucoarabin, glucobrassicanapin, glucobrassicin, glucocamelinin, glucoerucin, glucoiberin, gluconapin, gluconapoleiferin, gluconasturtiin, glucoraphanin, glucoraphenin, glucotropaeolin, homoglucocamelinin, 4-hydroxyglucobrassicin, 4-methoxyglucobrassicin, neoglucobrassicin, progoitrin, sinalbin and sinigrin.
The concentration ranges tested in the collaborative trial for each individual glucosinolate and for the total glucosinolate content are summarized in Table 1.
[table not represented]

This document specifies a method for the detection of hazelnut (Corylus avellana) in chocolate.
Real-time PCR (Polymerase chain reaction) detection of hazelnut is based on an152 bp (base pair) sequence from the corA 1 gene of hazelnut.

This document specifies the enumeration of sulfite-reducing Clostridium spp. by the colony-count technique.
This document is applicable to:
—    products intended for human consumption;
—    products for feeding animals;
—    environmental samples in the area of food and feed production and handling;
—    samples from the primary production stage.
NOTE      This method has been validated in an interlaboratory study for the following food categories:
—    ready-to-eat, ready-to-reheat meat products;
—    eggs and egg products (derivates);
—    processed fruits and vegetables;
—    infant formula and infant cereals;
—    multi-component foods or meal components.
It has also been validated for the following other categories:
—    pet food and animal feed;
—    environmental samples (food or feed production).
As this method has been validated for at least five food categories, this method is applicable for a broad range of food. For detailed information on the validation, see Clause 11 and Annex C. Since the method is not commonly used for samples in the primary production stage, this category was not included in the interlaboratory study. Therefore, no performance characteristics were obtained for this category.
This horizontal method was originally developed for the examination of all samples belonging to the food chain. Based on the information available at the time of publication of this document, this method is considered to be fully suited to the examination of all samples belonging to the food chain. However, because of the large variety of products in the food chain, it is possible that this horizontal method is not appropriate in every detail for all products. Nevertheless, it is expected that the required modifications are minimized so that they do not result in a significant deviation from this horizontal method.
This technique is suitable for, but not limited to, the enumeration of microorganisms in test samples with a minimum of 10 colonies counted on a plate. This corresponds to a level of contamination that is expected to be higher than 10 cfu/ml for liquid samples or higher than 100 cfu/g for solid samples.

This document specifies a method for the detection of peanut (Arachis hypogaea) in chocolate.
Real-time PCR (Polymerase Chain Reaction) detection of peanut is based on an 86 bp (base pair) sequence from the Ara h 2 gene of peanut.

This document specifies a method for the quantitative determination of pyrrolizidine alkaloids (PA) in the concentration ranges shown in Table 1 in complete and supplementary feed and in forages by liquid chromatography tandem mass spectrometry (LC-MS/MS) after solid phase extraction (SPE) clean-up.
Table 1 - Summary of concentration ranges per PA tested in the collaborative trial
NOTE 1   A second method was part of the method validation collaborative main trial. For this method PA-N-Oxides are reduced by adding zinc powder to the extract of the feed material. The following steps correspond to the first and main method. Quantitative results for each PA except the otonecine type PA senkirkine represent the sum of the free PA base and its corresponding N-oxide.
NOTE 2   Due to insufficient numbers of data for some analyte-matrix combinations statistical evaluation was not valid for standardization. Received data indicated the methods applicability in experienced laboratories with appropriate quality assurance measures. Therefore, the method description is included as an informative annex (Annex D).

This document specifies a procedure for the qualitative detection of species specific DNA from white mustard (Sinapis alba) and soya (Glycine max) in cooked sausages using singleplex real-time PCR based on the genes MADS-D (mustard) and lectin (soya).

This document specifies a method for the quantitative determination of pyrrolizidine alkaloids (PA) in the concentration ranges shown in Table 1 in complete and supplementary feed and in forages by liquid chromatography tandem mass spectrometry (LC-MS/MS) after solid phase extraction (SPE) clean-up.
Table 1 - Summary of concentration ranges per PA tested in the collaborative trial
NOTE 1   A second method was part of the method validation collaborative main trial. For this method PA-N-Oxides are reduced by adding zinc powder to the extract of the feed material. The following steps correspond to the first and main method. Quantitative results for each PA except the otonecine type PA senkirkine represent the sum of the free PA base and its corresponding N-oxide.
NOTE 2   Due to insufficient numbers of data for some analyte-matrix combinations statistical evaluation was not valid for standardization. Received data indicated the methods applicability in experienced laboratories with appropriate quality assurance measures. Therefore, the method description is included as an informative annex (Annex D).

This document specifies a method for the detection of hazelnut (Corylus avellana) in chocolate.
Real-time PCR (Polymerase chain reaction) detection of hazelnut is based on an152 bp (base pair) sequence from the corA 1 gene of hazelnut.

This document specifies a method for the detection of peanut (Arachis hypogaea) in chocolate.
Real-time PCR (Polymerase Chain Reaction) detection of peanut is based on an 86 bp (base pair) sequence from the Ara h 2 gene of peanut.

This document specifies a procedure for the qualitative detection of species specific DNA from white mustard (Sinapis alba) and soya (Glycine max) in cooked sausages using singleplex real-time PCR based on the genes MADS-D (mustard) and lectin (soya).

This document specifies the enumeration of sulfite-reducing Clostridium spp. by the colony-count technique.
This document is applicable to:
—    products intended for human consumption;
—    products for feeding animals;
—    environmental samples in the area of food and feed production and handling;
—    samples from the primary production stage.
NOTE      This method has been validated in an interlaboratory study for the following food categories:
—    ready-to-eat, ready-to-reheat meat products;
—    eggs and egg products (derivates);
—    processed fruits and vegetables;
—    infant formula and infant cereals;
—    multi-component foods or meal components.
It has also been validated for the following other categories:
—    pet food and animal feed;
—    environmental samples (food or feed production).
As this method has been validated for at least five food categories, this method is applicable for a broad range of food. For detailed information on the validation, see Clause 11 and Annex C. Since the method is not commonly used for samples in the primary production stage, this category was not included in the interlaboratory study. Therefore, no performance characteristics were obtained for this category.
This horizontal method was originally developed for the examination of all samples belonging to the food chain. Based on the information available at the time of publication of this document, this method is considered to be fully suited to the examination of all samples belonging to the food chain. However, because of the large variety of products in the food chain, it is possible that this horizontal method is not appropriate in every detail for all products. Nevertheless, it is expected that the required modifications are minimized so that they do not result in a significant deviation from this horizontal method.
This technique is suitable for, but not limited to, the enumeration of microorganisms in test samples with a minimum of 10 colonies counted on a plate. This corresponds to a level of contamination that is expected to be higher than 10 cfu/ml for liquid samples or higher than 100 cfu/g for solid samples.

This document describes a method for the determination of T-2 toxin and HT-2 toxin in cereals and cereal-based products, e.g. oats, intended for nutrition of infants and young children by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) after cleanup by solid phase extraction (SPE) [5].
The method has been validated for HT-2 toxin in oat flour at levels of 9,3 µg/kg and 28,1 µg/kg, oat flakes at levels of 16,5 µg/kg and 21,4 µg/kg, and breakfast cereals (containing oat flakes) at a level of 8,1 µg/kg and for T-2 toxin in oat flour at levels of 4,4 µg/kg and 8,3 µg/kg, oat flakes at levels of 4,9 µg/kg and 6,6 µg/kg and breakfast cereals (containing oat flakes) at a level of 3,5 µg/kg.
Laboratory experiences [6] have shown that the method is also applicable to highly swelling materials (dry cereal-based porridges and modified starches), but these were not examined in the method validation study. Details are outlined in 7.3.
The method can also be applied to oat-by-products at higher levels of T-2- and HT-2 toxin. In this case, the dilution steps need to be considered [6].
The method can also be applied to cereals and cereal products for infants and young children based on e.g. wheat, barley and rice. In this case, the method needs to be in-house-validated for each material. At the time of the interlaboratory study, planned range was 10 µg/kg to 100 µg/kg, and it is known from the pre-study that the method works well in the whole range, although final validation was only done in the range from 3,5 µg/kg to 28,1 µg/kg.

This document specifies the minimum requirements for generating and analysing whole genome sequencing (WGS) data of bacteria obtained from the food chain. This process can include the following stages:
a) handling of bacterial cultures;
b) axenic genomic DNA isolation;
c) library preparation, sequencing, and assessment of raw DNA sequence read quality and storage;
d) bioinformatics analysis for determining genetic relatedness, genetic content and predicting phenotype, and bioinformatics pipeline validation;
e) metadata capture and sequence repository deposition;
f) validation of the end-to-end WGS workflow (fit for purpose for intended application).
This document is applicable to bacteria isolated from:
—    products intended for human consumption;
—    products intended for animal feed;
—    environmental samples from food and feed handling and production areas;
—    samples from the primary production stage.

This document describes a method for the determination of T-2 toxin and HT-2 toxin in cereals and cereal-based products, e.g. oats, intended for nutrition of infants and young children by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) after cleanup by solid phase extraction (SPE) [5].
The method has been validated for HT-2 toxin in oat flour at levels of 9,3 µg/kg and 28,1 µg/kg, oat flakes at levels of 16,5 µg/kg and 21,4 µg/kg, and breakfast cereals (containing oat flakes) at a level of 8,1 µg/kg and for T-2 toxin in oat flour at levels of 4,4 µg/kg and 8,3 µg/kg, oat flakes at levels of 4,9 µg/kg and 6,6 µg/kg and breakfast cereals (containing oat flakes) at a level of 3,5 µg/kg.
Laboratory experiences [6] have shown that the method is also applicable to highly swelling materials (dry cereal-based porridges and modified starches), but these were not examined in the method validation study. Details are outlined in 7.3.
The method can also be applied to oat-by-products at higher levels of T-2- and HT-2 toxin. In this case, the dilution steps need to be considered [6].
The method can also be applied to cereals and cereal products for infants and young children based on e.g. wheat, barley and rice. In this case, the method needs to be in-house-validated for each material. At the time of the interlaboratory study, planned range was 10 µg/kg to 100 µg/kg, and it is known from the pre-study that the method works well in the whole range, although final validation was only done in the range from 3,5 µg/kg to 28,1 µg/kg.

This document specifies a method for the detection of foodstuff containing cellulose which have been treated with ionizing radiation, by analysing the electron spin resonance (ESR) spectrum, also called electron paramagnetic resonance (EPR) spectrum, of the foodstuff, see [1] to [13].
Interlaboratory studies have been successfully carried out with pistachio nut shells, [14] to [18], paprika powder [19] and [20] and fresh strawberries [21]. However, it has been shown that chemical bleaching of nuts in shells can lead to comparable signals. For further information, see Clause 8 on limitations.

This document specifies a method for the determination of free gossypol, extractable by acidified acetonitrile/water, in cottonseed, cottonseed products and compound feeds by liquid chromatography with tandem mass spectrometry (LC-MS/MS).
The method described in this document has been successfully validated in the range of 69 mg/kg to 5 950 mg/kg by collaborative trial in the following matrices: cottonseed, cottonseed products (cake/meal, hulls) and compound feeds for bovine, porcine and poultry.
NOTE   It is possible to reach quantification limits of approximatively 5 mg/kg in compound feeds. The method might be applicable at lower and at higher concentrations than the concentration range validated in the collaborative trial. However, this needs to be assessed by in-house validation.

This document specifies a method for the detection of foodstuff containing crystalline sugars which have been treated with ionizing radiation, by analysing the electron spin resonance (ESR) spectrum, also called electron paramagnetic resonance (EPR) spectrum, of the foodstuff, see [1] to [7].
Interlaboratory studies have been successfully carried out on dried figs, dried mangoes, dried papayas and raisins, see [1] to [3].

This document specifies the minimum requirements for generating and analysing whole genome sequencing (WGS) data of bacteria obtained from the food chain. This process can include the following stages:
a) handling of bacterial cultures;
b) axenic genomic DNA isolation;
c) library preparation, sequencing, and assessment of raw DNA sequence read quality and storage;
d) bioinformatics analysis for determining genetic relatedness, genetic content and predicting phenotype, and bioinformatics pipeline validation;
e) metadata capture and sequence repository deposition;
f) validation of the end-to-end WGS workflow (fit for purpose for intended application).
This document is applicable to bacteria isolated from:
—    products intended for human consumption;
—    products intended for animal feed;
—    environmental samples from food and feed handling and production areas;
—    samples from the primary production stage.

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.

This document specifies general rules for the enumeration of pediococci in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain pediococci as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see A.2). The document is not applicable to mineral feeds which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) 767/2009) [3]. There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

This document specifies general rules for the enumeration of enterococci (E. faecium) in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain enterococci as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see A.2). The document is not applicable to mineral feeds which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) 767/2009) [4].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

This document specifies general rules for the enumeration of bacilli in feeding stuffs (additives, premixtures and compound feeds including mineral feeds) [4] that contain bacilli as a single microorganism component or in a mixture with other microorganisms. There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets containing about 109 CFU/kg.

This document specifies general rules for the enumeration of lactobacilli in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain lactobacilli as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see A.2). The document is not applicable to mineral feeds, which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) No 767/2009) [3].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

This document specifies general rules for the enumeration of Saccharomyces cerevisiae in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain Saccharomyces cerevisiae as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see Annex A). The document is not applicable to mineral feeds, which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) 767/2009) [3].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

This document specifies a method for the determination of the content of the total vitamin A (retinol), vitamin E (α-tocopherol) and vitamin D3 (cholecalciferol) in animal feed using solid phase extraction (SPE) clean-up and high-performance liquid chromatography (HPLC).
NOTE   The procedure also enables determination of vitamin D2 but with the use of another internal standard. The method is fully validated only for vitamin D3.
The method has been successfully tested in collaborative trial for complete feed for broilers, pigs, and turkey, for premixture for broilers and piglets, for complementary feed for cows and mineral feed within the following ranges:
•   vitamin A: 4 365 IU/kg - 4 118 352 IU/kg;
•   vitamin E: 22 mg/kg - 13 800 mg/kg;
•   vitamin D3: 1 668 IU/kg - 1 638 150 IU/kg.
The limits of quantification were not determined within the validation study. Quantification limits of 1 100 IU for vitamin A/kg (using UV-detection), 4 mg for vitamin E/kg (using UV-detection), 2 mg for vitamin E/kg (using fluorescence detection) and 2 000 IU for vitamin D/kg (using UV-detection) should be normally achieved. Lower limits are possible provided they are validated by the user.

This document specifies a method for the detection of foodstuff containing crystalline sugars which have been treated with ionizing radiation, by analysing the electron spin resonance (ESR) spectrum, also called electron paramagnetic resonance (EPR) spectrum, of the foodstuff, see [1] to [7].
Interlaboratory studies have been successfully carried out on dried figs, dried mangoes, dried papayas and raisins, see [1] to [3].

This document specifies a method for the detection of foodstuff containing cellulose which have been treated with ionizing radiation, by analysing the electron spin resonance (ESR) spectrum, also called electron paramagnetic resonance (EPR) spectrum, of the foodstuff, see [1] to [13].
Interlaboratory studies have been successfully carried out with pistachio nut shells, [14] to [18], paprika powder [19] and [20] and fresh strawberries [21]. However, it has been shown that chemical bleaching of nuts in shells can lead to comparable signals. For further information, see Clause 8 on limitations.

This document specifies a method for the determination of free gossypol, extractable by acidified acetonitrile/water, in cottonseed, cottonseed products and compound feeds by liquid chromatography with tandem mass spectrometry (LC-MS/MS).
The method described in this document has been successfully validated in the range of 69 mg/kg to 5 950 mg/kg by collaborative trial in the following matrices: cottonseed, cottonseed products (cake/meal, hulls) and compound feeds for bovine, porcine and poultry.
NOTE   It is possible to reach quantification limits of approximatively 5 mg/kg in compound feeds. The method might be applicable at lower and at higher concentrations than the concentration range validated in the collaborative trial. However, this needs to be assessed by in-house validation.

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 on-line-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 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 on-line HPLC-GC-FID analysis that can be used as a screening method for the determination of MOSH.

This document specifies a method for the analysis of pesticide residues in foods of plant and of animal origin by ethyl acetate extraction using GC- and LC-MS/MS (SweEt).

This document specifies a method for the analysis of pesticide residues in foods of plant and of animal origin by ethyl acetate extraction using GC- and LC-MS/MS (SweEt).

This European Standard specifies a method for the determination of five Alternaria toxins in wheat, tomato juice and sunflower seed samples by liquid chromatography tandem mass spectrometry (LC-MS/MS). The method includes the analysis of Altenuene (ALT), Alternariol (AOH), Alternariol monomethyl ether (AME) in the range of 1 μg/kg to 100 μg/kg, and Tentoxin (TEN) in the range of 5 μg/kg to 500 μg/kg, and Tenuazonic acid (TEA) in the range of 10 μg/kg to 1000 μg/kg.

This document specifies general rules for the enumeration of enterococci (E. faecium) in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain enterococci as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see A.2). The document is not applicable to mineral feeds which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) 767/2009) [4].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

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.

This document specifies general rules for the enumeration of bacilli in feeding stuffs (additives, premixtures and compound feeds including mineral feeds) [4] that contain bacilli as a single microorganism component or in a mixture with other microorganisms. There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets containing about 109 CFU/kg.

This document specifies general rules for the enumeration of pediococci in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain pediococci as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see A.2). The document is not applicable to mineral feeds which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) 767/2009) [3]. There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

This document specifies general rules for the enumeration of lactobacilli in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain lactobacilli as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see A.2). The document is not applicable to mineral feeds, which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) No 767/2009) [3].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

This document specifies general rules for the enumeration of Saccharomyces cerevisiae in feeding stuffs (additives, premixtures and compound feeds excluding mineral feeds) that contain Saccharomyces cerevisiae as a single microorganism component or in a mixture with other microorganisms. Applying the method to premixtures and compound feeds with critical amounts of copper demands a special procedure (see Annex A). The document is not applicable to mineral feeds, which are defined as complementary feeding stuffs composed mainly of minerals and containing at least 40 % crude ash (Regulation (EC) 767/2009) [3].
There are different categories of feed samples:
a)   Additives containing about 1010 colony forming units (CFU)/g;
b)   Premixtures containing about 1011 CFU/kg;
c)   Compound feeds, meal or pellets which contain about 109 CFU/kg.

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 on-line-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 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 on-line HPLC-GC-FID analysis that can be used as a screening method for the determination of MOSH.