This document describes a procedure for the determination of aflatoxins B1, B2, G1 and G2 and total aflatoxins (sum of B1, B2, G1 and G2) in spices for which EU maximum levels are established, other than paprika, by high performance liquid chromatography (HPLC) with post-column derivatization (PCD) and fluorescence detection (FLD) after immunoaffinity column clean-up.
The method is applicable to the spices capsicum, pepper, nutmeg, ginger, turmeric and mixtures thereof.
The method has been val...
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    • Standard
      31 pages
      Slovenian language
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      33 pages
      English language
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The proposed deliverable specifies the procedure for single-laboratory validation of mainly non-proprietary methods in the fields of microbiological analysis of food, feed, and environmental and primary production stage samples. Single-laboratory validation is required if an interlaboratory validation according to ISO 16140-2 is not appropriate, e.g. for in-house methods or when the required number of participating laboratories is not available. Single-laboratory validation is not part of the op...view more

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    • Standard
      57 pages
      English language
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      47 pages
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The proposed deliverable specifies an alternative technical protocol for the validation of mostly non-proprietary methods in the field of microbiological analysis of food, animal feed, and environmental and primary production stage samples.
It is closely related to ISO 16140-2. The latter specifies the technical protocol for the validation of proprietary methods, including a classical interlaboratory study and a method comparison study to be conducted in one laboratory. The realization of class...
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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 internation...
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This document specifies performance criteria for the selection of single-laboratory validated or collaborative study validated methods of analysis of mycotoxins in feed. The terms and definition of the relevant parameters for method validation are included. The performance requirements and characteristics are provided. This document could serve as a guide:
— to assess the quality of new European Standard methods under validation;
— to review the quality of previous collaborative trials;
— to ...
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    • Technical specification
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    • Technical specification
      26 pages
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This document specifies a gas chromatographic method with electron capture detection (ECD) for the determination of organochlorine pesticides (OCP’s) in animal feeding stuffs.
The method is applicable to animal feeding stuffs with a water content up to about 20 % by weight and oil/fatty samples containing residues of one or more of the following OCP’s, toxaphene and some of their isomers and degradation products:
   aldrin;
   dieldrin;
   chlordane (as the sum of chlordane isomers and oxychlo...
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This document specifies a gas chromatographic mass spectrometric (GC/MS) method for the determination of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in animal feeding stuffs and oil.
The method is applicable to animal feeding stuffs consisting of less than 20 % by mass and oil/fatty samples containing residues of one or more of the following OCPs and PCBs and some of their isomers and degradation products:
   aldrin;
   dieldrin;
   chlordane, as the sum of chlordane ...
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      60 pages
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This document specifies a gas chromatographic method with electron capture detection (ECD) for the determination of organochlorine pesticides (OCPs) in compound feeds and oil and fats.
The method is applicable to animal compound feed, oils and fats and fish meals with a water content up to about 20 % by weight and oil/fatty samples containing residues of one or more of the following OCPs, toxaphene and some of their isomers and degradation products:
—   aldrin;
—   dieldrin;
—   dichlorodiphenyltr...
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      25 pages
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This document specifies a gas chromatographic mass spectrometric (GC/MS) method for the determination of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in animal feeding stuffs and oil.
The method is applicable to animal feeding stuffs consisting of less than 20 % by mass and oil/fatty samples containing residues of one or more of the following OCPs and PCBs and some of their isomers and degradation products:
   aldrin;
   dieldrin;
   chlordane, as the sum of chlordane ...
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      60 pages
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This document is applicable to the determination of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), (together termed ‘dioxins’ (PCDD/Fs)) and dioxin-like PCBs and non-dioxin-like PCBs (dl-PCBs and ndl-PCBs) in animal feeding stuffs. Collaborative studies have been carried out. The method is suitable for the determination of dioxins, dl-PCBs and ndl-PCBs at the appropriate MRL in compound feed and ingredients e.g. oil, mineral clay. The method is applicable to sa...view more

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    • Standard
      55 pages
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      54 pages
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This document describes a procedure for the determination of phomopsins in lupin seeds and lupin-derived products based on liquid chromatography with tandem mass spectrometry (LC-MS/MS). Several phomopsins exist, i.e. phomopsin A, B, C and D, but the method only deals with the quantitative measurement of phomopsin A due to lack of commercially available analytical reference standards for the other phomopsins.
The method has been validated for phomopsin A in naturally contaminated lupin seeds, ...
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This document describes a procedure for the determination of ochratoxin A (OTA) in chilli, paprika, black and white pepper, nutmeg, spice mix, liquorice (root and extracts), cocoa and cocoa products by high performance liquid chromatography (HPLC) with immunoaffinity column clean-up and fluorescence detection.
This method has been validated in interlaboratory studies via the analysis of both naturally contaminated and spiked samples ranging from 1,0 μg/kg to 84,9 μg/kg for spices (paprika and ...
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This document describes a procedure for the determination of ochratoxin A (OTA) in pork products specifically ham, pork based products (canned chopped pork) and pork liver using high performance liquid chromatography with fluorescence detection (HPLC-FLD).
The method has been validated for ochratoxin A with naturally contaminated ham, pork based products (canned chopped pork) and pork liver containing 0,5 μg/kg to 11 μg/kg [4, 5, 6].
Laboratory experiences have shown that this method is also ...
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This document specifies a procedure for the determination of ochratoxin A (OTA) in chilli, paprika, black and white pepper, nutmeg, spice mix, liquorice (root and extracts), cocoa and cocoa products by high performance liquid chromatography (HPLC) with immunoaffinity column clean-up and fluorescence detection (FLD).
This method has been validated in interlaboratory studies via the analysis of both naturally contaminated and spiked samples ranging from 1,0 μg/kg to 84,9 μg/kg for spices (paprika...
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      25 pages
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This document describes a procedure for the determination of ochratoxin A (OTA) in pork products specifically ham, pork-based products (canned chopped pork) and pork liver using high performance liquid chromatography with fluorescence detection (HPLC-FLD).
The method has been validated for ochratoxin A in naturally contaminated ham, pork based products (canned chopped pork) and pork liver containing 0,5 μg/kg to 11 μg/kg [4], [5], [6].
Laboratory experiences have shown that this method is also...
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This part of ISO 16140 specifies the general principle and the technical protocol for the validation of alternative, mostly proprietary, confirmation methods in the field of microbiological analysis of food, animal feed, and environmental and primary production stage samples. This procedure is limited to the validation of alternative (proprietary) confirmation methods that are intended to replace (partly or completely) the confirmatory procedure described in the standard method for the enumerati...view more

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    • Standard
      35 pages
      English language
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      30 pages
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This document is applicable to the determination of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), (together termed ‘dioxins’ (PCDD/Fs)) and dioxin-like PCBs and non-dioxin-like PCBs (dl-PCBs and ndl-PCBs) in animal feeding stuffs. Collaborative studies have been carried out. The method is suitable for the determination of dioxins, dl-PCBs and ndl-PCBs at the appropriate MRL in compound feed and ingredients e.g. oil, mineral clay. The method is applicable to sa...view more

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    • Standard
      55 pages
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      54 pages
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This document specifies a procedure for the determination of phomopsin A in lupin seeds and lupin-derived products based on liquid chromatography with tandem mass spectrometry (LC-MS/MS). Several phomopsins exist, i.e. phomopsin A, B, C and D, but the method only deals with the quantitative measurement of phomopsin A due to lack of commercially available analytical reference standards for the other phomopsins.
The method has been validated for phomopsin A in naturally contaminated lupin seeds, ...
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    • Standard
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      18 pages
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ISO 18862:2016 specifies methods for the determination of acrylamide in coffee and coffee products by extraction with water, clean-up by solid-phase extraction and determination by HPLC-MS/MS and GC-MS. It was validated in a method validation study on roasted coffee, soluble coffee, coffee substitutes and coffee products with ranges from 53 μg/kg to 612,1 μg/kg.

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    • Standard
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      24 pages
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This document describes a method for the determination of organomercury in seafood/fishery products by elemental mercury analysis. The method has been successfully valideted in an interlaboratory study with a working range from 0,013 mg/kg to 5,12 mg/kg (HORRAT values <2) in seafood/fishery products [1], [2]. The limit of quantification is approximately 0,010 mg/kg organomercury (referring to dry weight, expressed as mercury) [3], [4].
Organic species of mercury, other than monomethylmercury, a...
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    • Standard
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    • Standard
      15 pages
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      14 pages
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This International Standard gives requirements and guidance for the estimation and expression of measurement
uncertainty (MU) associated with quantitative results in microbiology of the food chain.
It is applicable to the quantitative analysis
 of products intended for human consumption or the feeding of animals, and
 of environmental samples in the area of food production and food handling,
 of samples at the stage of primary production.
The quantitative analysis is typically carried o...
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    • Standard
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      36 pages
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This European Standard method of analysis is applicable for the determination of Deoxynivalenol (DON) in the tested range of 96,2 µg/kg to 3 269 µg/kg, Aflatoxin B1 (AfB1) in the tested range of 2,62 µg/kg to 444 µg/kg, Fumonisin B1 (FB1) in the tested range of 693 µg/kg to 7 529 µg/kg, Fumonisin B2 (FB2) in the tested range of 203 µg/kg to 2 465 µg/kg, T-2 toxin in the tested range of 7,47 µg/kg to 360 µg/kg and HT-2 toxin in the tested range of 13,9 µg/kg to 1 758 µg/kg, Zearalenone (ZON) in t...view more

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    • Standard
      32 pages
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      33 pages
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This document method is applicable for the determination of theobromine in compound feed by liquid chromatography with UV detection in the tested range of 27 to 307 mg/kg. This method has been validated using complementary compound feed for adult dogs and complementary compound feedstuff for horses. The actual working range may extend beyond the tested range. Alternative chromatography conditions using liquid chromatography tandem mass spectrometry (LC-MS/MS) are also provided for the validated ...view more

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    • Standard
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      25 pages
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This document specifies the general principle and the technical protocol for the validation of alternative confirmation methods for microbiology in the food chain. This document compares the result of the alternative confirmation method against the confirmation procedure of a reference method or, if needed, a reference confirmation method (e.g. whole genome sequencing).
This document is applicable to the validation of alternative confirmation methods used for the analysis (detection or quantifi...
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    • Standard
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      30 pages
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This document specifies requirements and gives guidance for the estimation and expression of measurement uncertainty (MU) associated with quantitative results in microbiology of the food chain.
It is applicable to the quantitative analysis of:
— products intended for human consumption or the feeding of animals;
— environmental samples in the area of food production and food handling;
— samples at the stage of primary production.
The quantitative analysis is typically carried out by enumerat...
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      36 pages
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This document describes a method for the determination of individual ergot alkaloids and tropane alkaloids in unprocessed cereals and cereal-based compound feeds by high performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The ergot alkaloids covered in this method are: ergocornine, ergocorninine, ergocristine, ergocristinine, α-ergocryptine, α-ergocryptinine, ergometrine, ergometrinine, ergosine, ergosinine, ergotamine and ergotaminine. The tropane alkaloids cover...view more

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      39 pages
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This document describes a screening method for the determination of aflatoxin B1, deoxynivalenol, fumonisin B1 and B2, ochratoxin A, HT-2 and T-2 toxins, and zearalenone in foodstuffs by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS).
The aim of the screening method is to test compliance of foodstuff with regulatory limits or to determine whether a certain pre-defined level (the screening target concentration, STC) is exceeded or not. The result of t...
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This document specifies a method for the detection of celery (Apium graveolens) in emulsion-type sausages (e.g. Frankfurter, Wiener).
Real-time PCR detection of celery is based on an 101 bp (base pair) sequence from the gene of the mannitol dehydrogenase (GenBank Acc. No. AF067082 ) of celery (Apium graveolens).
The method has been validated on emulsion-type sausages (Bavarian “Leberkäse”) spiked with celery. For this purpose meat batter containing mass fractions of 50 % pork meat, 25 % pork ...
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This document provides an overall framework covering qualitative and quantitative methods for the determination of food allergens and allergenic ingredients using antibody-based methods in foods. This European Standard specifies general guidelines and performance criteria for antibody-based methods for the detection and quantification of proteins that serve as markers for the presence of allergy provoking foods or food ingredients. Other methods than those described can also detect and identify ...view more

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This document specifies how to use the standards for immunoassays, nucleic based and chromatographic methods and their relationship in the analysis of food allergens; and contains general definitions, requirements and guidelines for laboratory set-up, method validation requirements, description of methods, and test reports.
This document also specifies general guidelines for the requirements and use of reference materials for the determination of allergenic commodities in food products. The ter...
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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 purp...
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This document specifies minimum method performance requirements for enzyme-linked immunosorbent assays that quantify non-fragmented or fragmented gluten from wheat (e.g. Triticum aestivum), rye, and barley in raw and processed foodstuffs..

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This document provides the overall framework for detection of sequences corresponding to species containing allergens using the polymerase chain reaction (PCR). It relates to the requirements for the specific amplification of target nucleic acid sequences (DNA) and for the confirmation of the identity of the amplified nucleic acid sequence.
Guidelines, minimum requirements and performance criteria laid down in this document are intended to ensure that comparable and reproducible results are ob...
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This document describes a procedure for the determination of nivalenol (NIV), deoxynivalenol (DON) and its acetyl derivatives (3-acetyl-DON and 15-acetyl-DON), HT-2 and T-2 toxins (HT-2, T-2) and zearalenone (ZEA) in cereals and cereal products by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) after cleanup by solid phase extraction (SPE).
The method has been validated with both contaminated and spiked samples of wheat, wheat flour, and wheat cracker...
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      41 pages
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This document's method of analysis is applicable for the determination of:
-   deoxynivalenol (DON) in the tested range of 100 µg/kg to 3 300 µg/kg,
-   aflatoxin B1 (AfB1) in the tested range of 2,5 µg/kg to 440 µg/kg,
-   fumonisin B1 (FB1) in the tested range of 690 µg/kg to 7 500 µg/kg,
-   fumonisin B2 (FB2) in the tested range of 200 µg/kg to 2 500 µg/kg,
-   T-2 toxin in the tested range of 7,5 µg/kg to 360 µg/kg,
-   HT-2 toxin in the tested range of 14 µg/kg to 1 800 µg/kg,
-   zearalenone (ZE...
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      33 pages
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This document describes a method for the determination of organomercury in seafood/fishery products by elemental mercury analysis. The method has been successfully valideted in an interlaboratory study with a working range from 0,013 mg/kg to 5,12 mg/kg (HORRAT values <2) in seafood/fishery products [1], [2]. The limit of quantification is approximately 0,010 mg/kg organomercury (referring to dry weight, expressed as mercury) [3], [4].
Organic species of mercury, other than monomethylmercury, a...
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    • Standard
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      15 pages
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      14 pages
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ISO 18862:2016 specifies methods for the determination of acrylamide in coffee and coffee products by extraction with water, clean-up by solid-phase extraction and determination by HPLC-MS/MS and GC-MS. It was validated in a method validation study on roasted coffee, soluble coffee, coffee substitutes and coffee products with ranges from 53 μg/kg to 612,1 μg/kg.

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    • Standard
      27 pages
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      24 pages
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This document method is applicable for the determination of theobromine in compound feed by liquid chromatography with UV detection in the tested range of 27 to 307 mg/kg. This method has been validated using complementary compound feed for adult dogs and complementary compound feedstuff for horses. The actual working range may extend beyond the tested range. Alternative chromatography conditions using liquid chromatography tandem mass spectrometry (LC-MS/MS) are also provided for the validated ...view more

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    • Standard
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      25 pages
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This Technical Specification gives guidelines for the execution of calibration and quantitative evaluation of chromatographic procedures for the determination of pesticides and organic contaminants in residue analysis. In addition, the essential requirements for calibration are outlined.
The calibration of analytical procedures and the evaluation of analytical results need to be conducted according to uniform principles in order to allow for a comparison of analytical results (even from differe...
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    • Technical specification
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    • Technical specification
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This document describes a method for the determination of aluminium in food by inductively coupled plasma optical emission spectrometry (ICP-OES) after pressure digestion. This method is suitable for mass fraction in the range of 15 mg/kg to 200 mg/kg. At concentrations above 200 mg/kg digestion temperatures higher than 220 °C can be necessary to recover the aluminium as completely as possible.

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This document specifies a method for the determination of aluminium in food by inductively coupled plasma mass spectrometry (ICP-MS) after pressure digestion. This method is suitable for mass fractions in the range of 1 mg/kg to 200 mg/kg. At concentrations above 200 mg/kg digestion temperatures higher than 220 °C can be necessary to recover the aluminium as completely as possible.

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This Technical Specification describes a method for the analysis of pesticide residues in fatty oils of plant origin (essential oils are excluded). It has been validated in an interlaboratory test with olive oil. However, laboratory experiences have shown that this method is also applicable to other kinds of oils such as sunflower seed oil, sesame oil, flax seed oil, rape seed oil, grape seed oil, thistle oil and pumpkin seed oil.

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    • Technical specification
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    • Technical specification
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This document specifies a high-performance liquid chromatographic (HPLC) mass spectrometric (MS) method for screening and quantification of melamine in the concentration range between < 1 mg/kg and 100 mg/kg feed.
The method is validated in an international collaborative trial for melamine in complete feed, complementary feed, feed material, milk replacer and pet food including canned pet food in the range between 1 mg/kg and 80 mg/kg with particular regard to the by the European Commission est...
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      37 pages
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      36 pages
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This document describes a screening method for the determination of aflatoxin B1, deoxynivalenol, fumonisin B1 and B2, ochratoxin A, HT-2 and T-2 toxins, and zearalenone in foodstuffs by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS).
The aim of the screening method is to test compliance of foodstuff with regulatory limits or to determine whether a certain pre-defined level (the screening target concentration, STC) is exceeded or not. The result of t...
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      29 pages
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This document describes a method for the determination of individual ergot alkaloids and tropane alkaloids in unprocessed cereals and cereal-based compound feeds by high performance liquid chromatography with tandem mass spectrometry (LC-MS/MS).
This method has been successfully validated by collaborative trial in the following matrices: rye, barley, wheat, complete feed for bovine, porcine and poultry. Validation in buckwheat produced acceptable results, but the relative standard reproducibil...
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      39 pages
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This document specifies a procedure for the determination of nivalenol (NIV), deoxynivalenol (DON) and its acetyl derivatives (3-acetyl-DON and 15-acetyl-DON), HT-2 and T-2 toxins (HT-2 and T-2) and zearalenone (ZEN) in cereals and cereal products by high performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (MS/MS) after clean-up by solid phase extraction (SPE).
The method has been validated with samples of wheat, wheat flour, and wheat crackers. The wheat and the whea...
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This document specifies minimum method performance requirements for enzyme-linked immunosorbent assays that quantify non-fragmented or fragmented gluten from wheat (e.g. Triticum aestivum), rye, and barley in raw and processed foodstuffs.
This document is intended to be used in addition to EN 15842.

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      10 pages
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This document specifies a method for the detection of celery (Apium graveolens) in emulsion-type sausages (e.g. Frankfurter, Wiener).
Real-time PCR (polymerase chain reaction) detection of celery is based on an 101 bp (base pair) sequence from the gene of the mannitol dehydrogenase (GenBank Acc. No. AF067082 ) of celery (Apium graveolens).
The method has been validated on emulsion-type sausages (Bavarian “Leberkäse”) spiked with celery. For this purpose meat batter containing mass fractions of...
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This document specifies how to use the standards for immunoassays, nucleic based and chromatographic methods and their relationship in the analysis of food allergens; and contains general definitions, requirements and guidelines for laboratory set-up, method validation requirements, description of methods, and test reports.
This document also specifies general guidelines for the requirements and use of reference materials for the determination of allergenic commodities in food products. The ter...
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