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 and chili [5], black and white pepper, nutmeg and spice mix [6]), ranging from 7,7 μg/kg to 96,8 μg/kg for liquorice and liquorice products [7] and ranging from 2,1 μg/kg to 26,3 μg/kg for cocoa and cocoa products [6].
For further information on the validation, see Clause 10 and Annex B.

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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 applicable to pâté and kidney [4].

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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, lupin flour and crisp bread at levels ranging from approximately 5 µg/kg to 60 µg/kg

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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 quantification) of isolated microorganisms in:
— products intended for human consumption;
— products intended for animal feeding;
— environmental samples in the area of food and feed production, handling;
— samples from the primary production stage.
Validated alternative confirmation methods can be used to replace (partly or completely) the confirmation procedure described in:
— the reference method;
— an alternative method validated in accordance with ISO 16140-2 only if one of the isolation agars specified in the validation study of the alternative confirmation method is used.
This document is also applicable to the validation of alternative typing methods, where the reference method can be, for example, a serological method (e.g. serotyping of Salmonella) or a molecular method (e.g. typing of Shiga toxin-producing E. coli).
This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms, to be determined on a case-by-case basis.
Validation studies in accordance with this document are primarily intended to be performed by organizations or expert laboratories involved in method validation, but can also be used by a single laboratory, especially when performing in-house validation under certain conditions (see ISO 16140-4).

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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 enumeration of microorganisms using a colony-count technique. This document is also generally applicable to other quantitative analyses, including:
— most probable number (MPN) techniques;
— instrumental methods, such as impediometry, adenosine triphosphate (ATP) and flow cytometry;
— molecular methods, such as methods based on quantitative polymerase chain reaction (qPCR).
The uncertainty estimated by this document does not include systematic effects (bias).

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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, are also extracted and thus determined with this method. However, in seafood/fishery products the contribution from organic species of mercury other than monomethylmercury is negligible.

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This document specifies performance criteria for immunochemical methods for the detection and/or quantification of a specific protein or protein(s) of interest [POI(s)] in a specified matrix.
The methods discussed are applicable to the analysis of proteins from a variety of sample types. Some uses for these methods include, but are not limited to, analysing proteins involved in crop and food production, food processing, food marketing, food safety, biotechnology or disease indexing.

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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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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 the screening is either "negative" or "suspect". "Negative" (screen negative) means that the targeted mycotoxins are not detected or potentially present but below the STC. "Suspect" (screen positive) means that the established cut-off level is exceeded and the sample can contain one or more mycotoxins at a level higher than the STC.
For full identification and accurate quantification a second confirmatory quantitative analysis method is required which is outside the scope of this document.
The method is suitable for various types of foodstuff and has been validated for representative matrices from four commodity groups:
-   high starch and/or protein content and low water and fat content: wheat, cereal mixture, wheat flour and cornflakes;
-   high oil content: peanuts;
-   high sugar low water content: figs;
-   high water content: grape juice.
During validation, cut-off levels were established for the following screening target concentrations:
-   aflatoxin B1: 2 µg/kg to 5 µg/kg;
-   deoxynivalenol: 250 µg/kg to 865 µg/kg;
-   fumonisin B1: 200 µg/kg to 790 µg/kg;
-   fumonisin B2: 110 µg/kg to 230 µg/kg;
-   ochratoxin A: 4 µg/kg to 9 µg/kg;
-   T-2 toxin: 25 µg/kg;
-   HT-2 toxin: 25 µg/kg to 50 µg/kg;
-   zearalenone: 30 µg/kg to 100 µg/kg.

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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 wheat flour was prepared from a mixture of wheat and fungi infected wheat kernels. The wheat crackers were baked from wheat flour and water spiked with the target mycotoxins.
Validation levels for NIV ranged from 27,7 μg/kg to 378 μg/kg.
Validation levels for DON ranged from 234 μg/kg to 2420 μg/kg.
Validation levels for 3-acetyl-DON ranged from 18,5 μg/kg to 137 μg/kg.
Validation levels for 15-acetyl-DON ranged from 11,4 μg/kg to 142 μg/kg.
Validation levels for HT-2 ranged from 6,6 μg/kg to 134 μg/kg.
Validation levels for T-2 ranged from 2,1 μg/kg to 37,6 μg/kg.
Validation levels for ZEN ranged from 31,6 μg/kg to 230 μg/kg.
Laboratory experiences have shown that this method is also applicable to barley and oat flour, and rye based crackers [5], however, this has not been validated in a collaborative study.

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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 document 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 the proteins. Guidelines, minimum requirements and performance criteria laid down in this document are intended to ensure that reproducible results are obtained by different analysts in private and/or official control laboratories or when conducting onsite food testing.
This document is intended to be used in addition to EN 15842.
NOTE   This document could also be applicable to other sample types where the same principles for method validation and verification would apply.

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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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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 European Standards are intended to ensure that comparable and reproducible results are obtained in different laboratories. This document has been established for food matrices.
This document is intended to be used in addition to EN 15842.

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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 term "reference materials" in this document includes certified reference materials as well as quality control materials. Currently only a limited number of reference materials for food allergen determination are available. As new materials become accepted and validated, they can be appended as an annex to this document.
This document does not deal with sampling issues. It simply details processes involved from receipt of the laboratory sample to the end result.

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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 50 % pork meat, 25 % pork fat, 23 % crushed ice and 1,8 % of a mixture of sodium chloride, nitrite, nitrate, phosphates and ascorbates was prepared according to a standard procedure for emulsion-type sausage. The meat batter was spiked with either ground celery seeds or celery root powder to 1000 mg/kg. Lower spiking levels were obtained by diluting with celery-free meat batter. The batter was stuffed into casings and heated at 65 °C for 60 min [1].
This document is intended to be used in addition to EN 15842 and FprEN 15634 1.

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This document specifies a method for the determination of aluminium in food by inductively coupled plasma optical emission spectrometry (ICP-OES) after pressure digestion. This method was validated for wheat noodle, cheese, liver, beetroot and cocoa powder at mass fractions 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 was validated for infant formula, wheat noodle, cheese, liver, beetroot and cocoa powder at 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 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 different analytical procedures). They constitute the basis of any method validation and of the quality assurance within laboratories [1], [2], [3].
This Technical Specification does not consider issues of identification/qualification and extraction efficiency.

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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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This document provides information on how the performance characteristics of qualitative (binary) real-time polymerase chain reaction (PCR) methods for detection of specific DNA sequences present in foods should be evaluated and validated by conducting a collaborative study.
The guidelines are applicable for validation of qualitative PCR methods used for detection of DNA sequences derived from genetically modified foodstuffs. They can be applicable also for PCR methods used for detection of other target sequences in foodstuffs, e.g. for species detection and identification.

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This document describes a procedure for the identification of single fish and fish fillets to the level of genus or 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. The methodology allows the identification of a large number of commercially important fish species.
The decision whether the cytb or cox1 gene segment or both are used for fish identification depends on the declared fish species, the applicability of the PCR method for the fish species and the availability of comparative sequences in the public databases.
This method has been successfully validated on raw fish fillets, however, laboratory experience is available that it can also be applied to processed, e.g. cold smoked, hot smoked, salted, frozen, cooked, fried, deep-fried samples.
This document 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 is not applicable for complex fish products containing mixtures of two or more fish species.

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This European Standard stipulates a method for the analysis of pesticide residues in foods of plant origin, such as fruits (including dried fruits), vegetables, cereals and many processed products thereof by using GC, GC-MS(/MS), and/or LC-MS(/MS). The method has been collaboratively studied on a large number of commodity/pesticide combinations. Precision data are summarized in FprCEN/TR 17063. Guidelines for calibration are outlined in FprCEN/TS 17061.

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This European Standard specifies a high-performance liquid chromatographic (HPLC) and an ion chromatographic (IC) method for determination of the nitrate level in vegetables and vegetable products. This method is applicable for samples with a content of 25 mg/kg or greater.
It has been validated on naturally contaminated and spiked samples as beetroot juice with nitrate mass fractions of 194 mg/kg and 691 mg/kg, pureed carrots with nitrate mass fractions of 26 mg/kg and 222 mg/kg and with iceberg lettuce with nitrate mass fractions of 623 mg/kg and 3 542 mg/kg.

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This European Standard specifies a highly efficient method for the determination of saturated and aromatic hydrocarbons (from C10 to C50) in vegetable fats and oils and foodstuff on basis of vegetable oils for which it has been interlaboratory validated, with online-HPLC-GC-FID [1], [2] and [3]. This standard 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 has been tested in an interlaboratory study via the analysis of both naturally contaminated and spiked vegetable oil samples and mayonnaise and margarine samples, ranging from 4 mg/kg to 197 mg/kg for MOSH, and from 2 mg/kg to 51 mg/kg for MOAH.
According to the results of the interlaboratory studies, the method has been proven suitable for MOSH- and MOAH mass concentrations each above 10 mg/kg.
In case of suspected interferences from natural sources, the mineral origin of the MOSH and MOAH fraction can be verified by examination of the pattern by GC-MS.

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This Technical Specification specifies a method for the determination of acrylamide in cereal-based products, potato-based products and coffee by gas-chromatography mass spectrometry (GC-MS).
The method has been single-laboratory validated via the analysis of spiked samples (French fries (uncooked), bread, water biscuit, infant cereal, biscuit, green coffee, roast coffee and instant coffee), ranging from 30 μg/kg to 1 500 μg/kg acrylamide.
The results from the single laboratory validation were obtained by a laboratory with significant experience in acrylamide analysis. In addition, this method has also been studied by inter laboratory trial via the analysis of samples containing incurred acrylamide, ranging from approximately 200 μg/kg to 2 000 μg/kg. Critical points of the method are identified in 7.5 and Clause 8.

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This European Standard 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 6.3.
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.

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This European Standard specifies a method for the determination of melamine and cyanuric acid in foodstuffs with liquid chromatography in combination with tandem mass spectrometry. The method has been validated in an interlaboratory study via the analysis of spiked samples of milk based infant formula, soy based infant formula, milk powder, whole milk, soy drink and milk chocolate ranging from 0,71 mg/kg to 1,43 mg/kg for melamine and 0,57 mg/kg to 1,45 mg/kg for cyanuric acid. The limits of quantification (LOQ) for melamine and cyanuric acid in food are 0,05 mg/kg and 0,25 mg/kg, respectively. The upper limit of the working range is up to 10 mg/kg for melamine and up to 25 mg/kg for cyanuric acid.

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This European Standard describes a procedure for the determination of the zearalenone content in edible vegetable oils specifically maize germ oil by either of the following techniques: High performance liquid chromatography with fluorescence detection (LC-FLD) or high performance liquid chromatography with tandem mass spectrometry (LC-MS/MS) after basic extraction of the diluted oil.
The method has been validated for zearalenone in naturally contaminated maize germ oil at levels of 61,2 µg/kg to 515 µg/kg [5].
Laboratory experiences [6] have shown that this method is also applicable to other vegetable oils such as wheat germ oil (n = 4), sunflower oil (n = 5), pumpkin seed oil (n = 1), soybean oil (n = 5), hemp seed oil (n = 5), rape seed oil (n = 11), and mixed oils including maize germ oil (n = 3). However occasionally, samples can result in interferences in the FLD-chromatograms. In this case, the detection with MS/MS is recommended.

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This European Standard specifies a gas chromatographic method using mass spectrometric detection for the determination of ethyl carbamate (EC) in stone fruit spirits, fruit marc spirits and other spirit drinks.
The method has been validated in an interlaboratory study for stone fruit spirits and fruit liqueurs, at levels ranging from 0,253 mg/l to 1,11 mg/l. However, linearity of the instrument response was proven for the concentration ranges 0,10 mg/l to 4,0 mg/l (simplified method) and 0,025 mg/l to 3,0 mg/l (procedure including sample clean-up), respectively.

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This European Standard describes a method for the determination of minerals and trace elements in foodstuffs using optical emission spectrometry with inductively coupled plasma (ICP-OES) after pressure digestion.
This method has been validated in an interlaboratory study according to ISO 5725 [1] on children's food soya, cheese, chicken meat, wheat flour, apple juice, lobster and milk, with calcium ranging from 70 mg/kg to 7178 mg/kg, with copper ranging from 0,60 mg/kg to 16,40 mg/kg, with iron ranging from 0,88 mg/kg to 77 mg/kg, with potassium ranging from 605 mg/kg to 14 312 mg/kg, with magnesium ranging from 45 mg/kg to 1 174 mg/kg, with manganese ranging from 0,44 mg/kg to 5,12 mg/kg, with sodium ranging from 11 mg/kg to 2 220 mg/kg, with phosphorus ranging from 72 mg/kg to 9 708 mg/kg, with sulfur ranging from 26 mg/kg to 8 542 mg/kg and with zinc ranging from 0,16 mg/kg to 43,5 mg/kg.

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This European Standard specifies a method for the determination of benzene in soft drinks, other beverages and vegetable-based infant foods, by headspace gas chromatography mass spectrometry (HS-GC-MS). The method has been validated in an interlaboratory study via the analysis of spiked samples of carbonated soft drink, still fruit-based drink, carbonated fruit-based drink, vegetable and fruit juice containing carrot, infant food vegetable based and infant food containing meat, ranging from 1,9 µg/kg to 18,6 µg/kg. However, linearity of the instrument response was proven for the concentration range from 0,5 µg/kg to 20 µg/kg. The limit of quantification (LOQ) depends on the instrument but can generally be expected to be in the range from 0,5 µg/kg to 1,0 µg/kg.

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This Technical Report lists the validation data which were obtained with EN 15662:2008 and prEN 15662:2016 in interlaboratory tests and in single laboratory method validation studies.

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This European standard specifies a method [1] for the quantitative determination of saxitoxin (STX), decarbamoyl saxitoxin (dcSTX), neosaxitoxin (NEO), decarbamoyl neosaxitoxin (dcNEO), gonyautoxin 1 and 4 (GTX1,4; sum of isomers), gonyautoxin 2 and 3 (GTX2,3; sum of isomers), gonyautoxin 5 (GTX5 also called B1), gonyautoxin 6 (GTX6 also called B2), decarbamoyl gonyautoxin 2 and 3 (dcGTX2,3; sum of isomers), N-sulfocarbamoyl-gonyautoxin 1 and 2 (C1,2; sum of isomers) and (depending on the availability of certified reference materials (CRMs)) N-sulfocarbamoyl-gonyautoxin 3 and 4 (C3,4; sum of isomers) in (raw) mussels, oysters, scallops and clams. Laboratory experience has shown that it is also be applicable in other shellfish [2], [3] and cooked shellfish products. The method described was validated in an interlaboratory study [4], [5] and was also verified in a EURL-performance test aiming the total toxicity of the samples [6]. Toxins which were not available in the first interlaboratory study [4], [5] as dcGTX2,3 and dcNEO were validated in two additional interlaboratory studies [7], [8]. The lowest validated levels [4], [5], [8], are given in µg toxin (free base) per kg shellfish tissue and also as µmol/kg shellfish tissue and are listed in Table 1.
A quantitative determination of GTX6 (B2) was not included in the first interlaboratory study but several laboratories detected this toxin directly after solid phase extraction with ion-exchange (SPE-COOH) clean-up and reported a mass concentration of 30 µg/kg or higher in certain samples. For that reason, the present method is applicable to quantify GTX6 (B2) directly, depending on the availability of the standard substance. Currently it is possible to determine GTX6 after a hydrolysis of Fraction 2 of the SPE-COOH clean-up, described in 6.4 as NEO. The indirect quantification of GTX6 was validated in two additional interlaboratory studies [7], [8].
A quantitative determination of C3,4 was included in the first interlaboratory study. The present method is applicable to quantify C3,4 directly, depending on the availability of the standard substance. If no standard substances are available, C3,4 can only be quantified as GTX1,4 if the same hydrolysis protocol used for GTX6 (6.4) is applied to Fraction 1 of the SPE-COOH clean-up, see [10].

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This European Standard specifies methods for the quantitative determination of domoic acid in raw bivalve molluscs and finfish as well as in cooked mussels. The limit of detection is about 10 ng/ml to 80 ng/ml (0,05 mg/kg to 0,4 mg/kg), depending on the UV detector sensitivity. The limit of quantification for DA by these methods is at least 2,7 mg/kg. Method A has been validated for the determination of DA in different raw matrices such as mussels, clams, scallops and anchovies, spiked and/or naturally contaminated at levels ranging from 2,7 mg/kg to 85,1 mg/kg. Method B has been validated for the determination of DA at levels ranging from 5 mg/kg to 12,9 mg/kg in cooked blue mussels.
For further information on validation data, see Clause 8 and Annex A.
Laboratory experience has shown that this standard can also be applied to other shellfish species, however, no complete validation study according to ISO 5725 has been carried out so far.

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This Technical Specification 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) [1]. A mustard content of 10 mg/kg or greater and a soya content of 10 mg/kg or greater can be detected with a probability of > 95 %.

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This Technical Specification describes a procedure for the qualitative detection of hazelnut (Corylus avellana) in chocolate. DNA is extracted from the chocolate and a specific DNA sequence for hazelnut detected from the gene for corA 1 [4], [5].

  • Technical specification
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This draft European Standard describes a procedure for the determination of inorganic arsenic in foodstuffs of marine and plant origin by anion-exchange HPLC-ICP-MS following waterbath extraction.
This method has been validated in an interlaboratory test on white rice, wholemeal rice, leek, blue mussels, fish muscle and seaweed with an inorganic arsenic mass fraction in the range 0,073 mg/kg to 10,3 mg/kg [1].

  • Standard
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This final draft European Standard describes a method for the determination of monomethylmercury (MMHg) in foodstuffs of marine origin. The method has been validated in an interlaboratory test on mussel tissue, squid muscle, crab claw muscle, dog fish liver, whale meat, cod muscle and Greenland halibut muscle (all freeze-dried) at levels from 0,04 mg/kg to 3,6 mg/kg dry weight according to ISO 5725 2 [1].
Laboratory experiences have shown that this method is also applicable on fresh samples [2].

  • Standard
    14 pages
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This Technical Specification describes a procedure for the qualitative detection of peanut (Arachis hypogaea) in chocolate using real-time PCR based on the gene for the peanut allergen Ara h 2 [1], [2].

  • Technical specification
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This European Standard specifies a method for the determination of fumonisin B1 (FB1) and fumonisin B2 (FB2) in processed maize-containing foods for infants and young children by high performance liquid chromatography (HPLC) with immunoaffinity cleanup and fluorescence detection (FLD). This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples ranging from 112 µg/kg to 458 µg/kg for FB1+FB2, 89 µg/kg to 384 µg/kg for FB1 and 22 µg/kg to 74 µg/kg for FB2.
For further information on the validation, see Clause 8 and Annex B.

  • Standard
    19 pages
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This European Standard specifies a method for the determination of furan in coffee and coffee products with headspace-gas chromatography-mass spectrometry (HS-GC-MS), see [1] and [2]. Coffee products in the scope of this method are extracts which have been spray-dried, agglomerated or freeze-dried. The method has been validated in an interlaboratory study via the analysis of naturally contaminated samples of spray-dried coffee, freeze-dried coffee and ground roasted coffee ranging from 264 µg/kg to 2 840 µg/kg.

  • Standard
    15 pages
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This European Standard specifies a method for the determination of 4 of the 16 EU priority polycyclic aromatic hydrocarbons (PAHs), identified as target PAHs. They are benz[a]anthracene (BaA), benzo[a]pyrene (BaP), benzo[b]fluoranthene (BbF) and chrysene (CHR). The method allows their quantification in the presence of the other 12 EU priority PAHs (benzo[j]fluoranthene (BjF), cyclopenta[cd]pyrene (CPP), benzo[k]fluoranthene (BkF), dibenz[a,h]anthracene (DhA), benzo[c]fluorene (BcL), dibenzo[a,e]pyrene (DeP), benzo[ghi]perylene (BgP), dibenzo[a,h]pyrene (DhP), dibenzo[a,i]pyrene (DiP), dibenzo[a,l]pyrene (DlP), indeno[1,2,3-cd]pyrene (IcP), 5-methylchrysene (5MC)) in extruded wheat flour, smoked fish, dry infant formula, sausage meat, freeze-dried mussels, edible oil and wheat flour, by gas-chromatography mass-spectrometry (GC-MS). The extraction of PAHs from solid samples is performed by pressurized liquid extraction (PLE). Soxhlet extraction was applied by some participants in the validation study by collaborative trial as alternative to PLE. The sample cleanup is performed by applying the following techniques in the reported sequence: size exclusion chromatography (SEC), and solid phase extraction (SPE).
This method complies with the performance characteristics specified in Commission Regulation (EU) No 836/2011 (see [1]). In particular the specifications for the limit of detection (LOD) and of the limit of quantification (LOQ) (0,30 µg/kg and 0,90 µg/kg respectively) were met.
The method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples, ranging from 0,5 µg/kg to 11,9 µg/kg. However, linearity of the instrument response was proven for the concentration range 0,5 µg/kg to 20 µg/kg.
For the determination of PAHs in edible fats and oils, two other CEN standards are also available, EN ISO 22959 and EN ISO 15753, for more information see [2] and [3].

  • Standard
    37 pages
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This European Standard specifies a method for the determination of acrylamide in bakery ware such as bread, toasted bread, crisp bread, butter cookies, and biscuits, as well as potato products such as potato chips, potato crisps, and potato pan cake and roasted coffee, by liquid chromatography in combination with electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS). This method has been validated in an interlaboratory study via the analysis of both naturally contaminated and spiked samples, ranging from 14,3 µg/kg to 9 083 µg/kg. It was developed at the Swedish National Food Administration and validated in a study organized by the Directorate General Joint Research Centre (DG JRC), Swedish National Food Administration and the Nordic Committee on Food Analysis (NMKL), see [1] and [2].
The limit of quantification (LOQ) depends on the type of instrument used and on the actual performance of the instrument. The majority of the laboratories participating in the validation study were able to determine acrylamide in a butter cookie sample at a level of 14,3 µg/kg. Thus, the validation by interlaboratory study showed that LOQ can be expected to be in the range between below 15 µg/kg and 30 µg/kg.

  • Standard
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This Technical Specification describes a screening procedure for the determination of nitric-acid extractable inorganic arsenic in rice with hydride generation-AAS.
The method has been developed and validated for the application of analysis in rice. It has been validated in an interlaboratory study according to ISO 5725 [2] on parboiled rice and brown rice having an inorganic arsenic content of 0,092 mg/kg and 0,191 mg/kg.

  • Technical specification
    12 pages
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This European Standard specifies a method for the pressure digestion of foodstuffs intended for the determination of elements. This method has been collaboratively tested in combination with atomic absorption (flame, electrothermal (ET), hydride, cold-vapour) techniques and ICP-MS. Other techniques such as e.g. ICP-OES, voltammetry or atomic fluorescence can be used in combination with this European Standard.

  • Standard
    12 pages
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This Technical Specification describes screening strategies for the detection of genetically modified (GM) DNA in food products by means of PCR methods. The strategies have been established for food matrices, but it can also be applied to other matrices (e.g. feed, seed and samples from field grown plants).
Detection of GM DNA is based on PCR methods targeting segments of transgenic DNA sequences (genetic elements, genetic constructs or insertion sites of transgenes). Various combinations of these PCR methods are involved in screening strategies. The methods are applied simultaneously or hierarchically. The general strategy is based on the matrix approach. Examples for the implementation and application of this approach are described.
In order to ensure reliable analytical results, the document also provides guidelines for the validation of the performance criteria of qualitative PCR methods applied in screening approaches.

  • Technical specification
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This Technical Report lists the mass spectrometric parameters which are useful for the application of European Standards for the determination of pesticide residues in foods of plant origin that use GC-MS. These European Standards are as follows:
EN 1528 (all parts), Fatty food - Determination of pesticides and polychlorinated biphenyls (PCBs)
EN 12393 (all parts), Foods of plant origin  - Multiresidue methods for the gas chromatographic determination of pesticide residues
EN 15662, Foods of plant origin  - Determination of pesticide residues using GC-MS and/or LC-MS/MS following acetonitrile extraction/partitioning  - QuEChERS-method
To facilitate the determination of pesticides and/or metabolites using GC-MS/MS, Table 2 specifies the diagnostic ion transitions suitable for identification and quantification, which can be used.

  • Technical report
    17 pages
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This European Standard specifies a method for the determination of vitamin B2 in food by high performance liquid chromatography (HPLC) and fluorescence detection. This method has been validated in two interlaboratory studies. The first study was for the analysis of samples of milk powder and pig's liver ranging from 1,45 mg/100 g to 10,68 mg/100 g. The second study was for the analysis of samples of tube feeding solution, baby food, powdered milk, meal with fruits, yeast, cereal and chocolate powder ranging from 0,21 mg/100 g to 87,1 mg/100 g. Vitamin B2 is the mass fraction of total riboflavin including its phosphorylated derivatives.
For further information on the validation, see Clause 8 and Annex B.

  • Standard
    15 pages
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This European Standard specifies a method for the determination of vitamin B6 in foodstuffs by high performance liquid chromatography (HPLC). Vitamin B6 is the mass fraction of the sum of pyridoxine, pyridoxal, pyridoxamine including their phosphorylated derivatives determined as pyridoxine. The β-glycosylated forms are not taken into account. These can be determined with the method given in EN 14663 [1] by which the different vitamins of vitamin B6 (pyridoxal, pyridoxamine and pyridoxine) are separated and individually quantified. A third European Standard, EN 14166 [2], determines the total vitamin B6 by microbiological assay.

  • Standard
    16 pages
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This European Standard specifies a method for the determination of vitamin E in foods by high performance liquid chromatography (HPLC). The determination of vitamin E content is carried out by measurement of α-, β , γ- and δ-tocopherol. This method has been validated in two interlaboratory studies. The first study was for the analysis of α-tocopherol in margarine and milk powder ranging from 9,89 mg/100 g to 24,09 mg/100 g. The second study was for the analysis of α-, β-, γ- and δ-tocopherol in milk powder and of α-, and β-tocopherol in oat powder ranging from 0,057 mg/100 g (β-tocopherol) to 10,2 mg/100 g (α-tocopherol).
NOTE   The vitamin E activity can be calculated from the tocopherol content assuming appropriate factors as given in [1], [2], [3] and [4].

  • Standard
    20 pages
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