KŽP - Agricultural food products

This document specifies a procedure for determining hydroxyacyl-coenzyme A dehydrogenase (HADH) activity in poultry meat samples by spectrophotometry. The results provide an indication of whether poultry breast meat has been previously frozen, based on the ratio of relative HADH activity, and can be used to verify the labelling of poultry breast meat sold as chilled poultry.
This document is applicable specifically to chicken breast meat. It is not applicable to other cuts and/or types of meat unless these have been validated and appropriate limits have been established.
When following the procedure outlined in this document, for a single sample pressing results in 96,6 % of samples being correctly classified. If a correct classification rate of 99,9 % is required, three sample pressings are necessary; i.e. three chicken breasts (one pressing from each), selected randomly from a batch.
This document is not applicable to minced meat or to poultry preparations.

This document specifies a method for determining the content of wet gluten and the gluten index for wheat flours (Triticum aestivum L. and Triticum durum Desf.) by mechanical means.
This document is directly applicable to flours. It is also applicable to common and durum wheat after grinding, if their particular size distribution meets the specification given in Table B.1.
NOTE            This document is related to ICC 137/1[7], ICC 155[8] and AACC Method 38-12.02[9].

This document specifies procedures for the detection of a DNA sequence of a construct used to (genetically) enhance the growth of fish commonly found in aquaculture. The genetically modified AquAdvantage Atlantic salmon (Salmo salar) carries the construct expressing CS-GHc2 growth hormone and can be detected based on a real-time polymerase chain reaction (PCR) targeting either the border between the growth hormone coding sequence (CS-GHc2) of Oncorhynchus tshawytscha (Chinook salmon) and the antifreeze terminator (T-AFP) of (Macro-) Zoarces americanus (ocean pout), i.e. with the construct-specific method, or the border between the Atlantic salmon genomic DNA and the antifreeze promoter (P-AFP) of ocean pout, i.e. with the event-specific method. These methods can be applied to identify the genetically modified (GM) fish or for screening purposes.
This document is applicable for the analysis of DNA extracted from foodstuffs. It can also be suitable for the analysis of DNA extracted from other products such as feedstuffs. The application of these methods requires the extraction of an adequate amount of amplifiable DNA from the relevant matrix.

This document specifies a procedure for determining whether a perceptible sensory difference or similarity exists between samples of two products. The method is a forced-choice procedure. The method is applicable whether a difference exists in a single sensory attribute or in several attributes.
The method is statistically less efficient than the triangle test (described in ISO 4120) but is easier to perform by the assessors.
The method is applicable even when the nature of the difference is unknown (i.e. it determines neither the size nor the direction of difference between samples, nor is there any indication of the attribute(s) responsible for the difference). The method is applicable only if the products are fairly homogeneous.
The method is effective for:
determining that either:
a perceptible difference results (duo-trio testing for difference); or
a meaningful perceptible difference does not result (duo-trio testing for similarity) when, for example, a change is made in ingredients, processing, packaging, handling or storage;
selecting, training and monitoring assessors.
Two forms of the method are described:
the constant-reference technique, used when one product is familiar to the assessors (e.g. a sample from regular production);
the balanced-reference technique, used when one product is not more familiar than the other.

This document specifies a method of preparing a ground sample of spice or condiment for analysis, from a laboratory sample obtained by the method specified in ISO 948.
This document is applicable to the majority of spices and condiments. However, in view of the large number and diversity of spices and condiments, it can be necessary in certain special cases, for example, considerable hardness, or high moisture, volatile oil or fat content, to use a modified procedure or to choose another more
suitable method. Any such modified procedure or alternative method will be indicated in the International Standard appropriate to the spice or condiment concerned.

This document 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), gonyautoxin 6 (GTX6), decarbamoyl gonyautoxin 2 and 3 (dcGTX2,3; sum of isomers), N-sulfocarbamoyl gonyautoxin 2 and 3 (C1,2; sum of isomers) and N-sulfocarbamoyl gonyautoxin 1 and 4 (C3,4; sum of isomers) in (raw) mussels, oysters, scallops and clams. Laboratory experience has shown that this document can also be applied to other marine invertebrates [2], [3] and processed products of those species, however, no complete interlaboratory validation study according to ISO 5725-2 [21] has been carried out so far. The method described was validated in an interlaboratory study [4], [5] and was also verified in a European Union Reference Laboratory for Marine Biotoxins (EURLMB)-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 as mass fraction of toxin (free base) in µg/kg shellfish tissue and also as µmol/kg shellfish tissue and are listed in Table 1.
[Table 1 - Lowest validated levels]
A quantitative determination of GTX6 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 fraction (free base) of 30 µg/kg or higher in certain samples. For that reason, the present method is applicable to quantify GTX6 directly, depending on the availability of the standard substance. Whenever GTX6 standard is not commercially available, it is possible to determine GTX6 after hydrolysis of Fraction 2 of the SPE-COOH clean-up, described in 7.4, as NEO. The indirect quantification of GTX6 was validated in two additional interlaboratory studies [7], [8]. A study to compare direct and indirect GTX6 quantification was conducted at the EURLMB [16].
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 (7.4) is applied to Fraction 1 of the SPE-COOH clean-up [10]. A study to compare direct and indirect C3,4 quantification was conducted at the EURLMB [16].

This document specifies a method for the determination of fatty acid methyl esters (FAMEs) derived by transesterification or esterification from fats, oils, and fatty acids by capillary gas chromatography (GLC). FAMEs from C4 to C24 can be separated using this document including saturated FAMEs, cis- and trans-monounsaturated FAMEs, and cis- and trans-polyunsaturated FAMEs.
This document is applicable to crude, refined, partially hydrogenated or fully hydrogenated fats, oils and fatty acids derived from animal and vegetable sources, and fats extracted from foodstuff.
This document does not apply to milk and milk products (or fat coming from milk and milk products) or products supplemented with conjugated linoleic acid (CLA).
This document does not apply to di-, tri-, polymerized, hydroxylated and oxidized fatty acids, and fats and oils.
A method for the determination of the composition of FAMEs expressed by area % in liquid vegetable oils is proposed in Annex E.

This document specifies a method for the selective enumeration of bifidobacteria in milk products by using a colony-count technique at 37 °C under anaerobic conditions.
The method is applicable to milk products, such as fermented (e.g. yoghurts) and non-fermented milks (e.g. pasteurized milks, skim milks, whey protein concentrates), milk powders and formulae (e.g. infant formulae, follow-up formulae for older infants, products for young children) where these microorganisms are present and viable, in combination with other lactic acid bacteria or alone. The method is also applicable to starter and probiotic cultures. For proposed quality criteria of dairy products, see, for example, CXS 243-2003.
Bifidobacteria used in milk products usually belong to the following species (e.g. References [7] and [10]):
—     Bifidobacterium adolescentis;
—     B. animalis subsp. animalis;
—     B. animalis subsp. lactis;
—     B. bifidum;
—     B. breve;
—     B. longum subsp. infantis;
—     B. longum subsp. longum.

This document specifies a method for the analysis of pesticide residues in foods of animal origin with a low fat content, such as meat/muscle, egg or milk by LC-MS/MS. Because of the low material requirements for miniaturized processing and the few work steps, the process is particularly time and cost-saving with high reliability and effectiveness. The method has been collaboratively studied on a number of commodity/pesticide combinations. Precision data are summarized in Table B.1. Guidelines for calibration are outlined in CEN/TS 17061:2019.

This document establishes basic principles and specifies requirements and methods to determine the cardinal values of bacteria and yeast strains and use them to predict microbial growth.
The four main steps of the approach are:
determination of the cardinal values in culture medium;
determination of the correction factor in the target food;
validation of the model;
simulations.
Four environmental factors are considered: temperature, pH, aw and inhibitors (e.g. organic acids).
NOTE 1        Microbial competition is not considered as an inhibitor in this document and can be addressed by proper modelling approaches.
The determination of cardinal values is performed in a two-step approach:
the determination of maximum specific growth rates of the studied strain grown in broth under a defined range of values of the studied environmental factor(s);
the use of recognized predictive microbiology secondary models to fit the obtained experimental data to obtain the cardinal values.
The use of cardinal values in microbial growth simulation is based on predictive microbiology primary and secondary models. The cardinal values are combined with challenge test data to consider the matrix effect. Depending on the goal of the growth simulation, it is important to account for variation of cardinal values between strains within a bacterial or yeast species.
Cardinal values are a good indicator of a strain growth ability for the studied environmental factors. They are therefore used as criteria to select strains, in addition to their origin and virulence, when performing growth challenge tests (see ISO 20976-1) or in methods validation (see ISO 16140 series).
NOTE 2        This document focuses on the determination of cardinal values for one strain. The same methodology can be used to characterize multiple strains independently to cover biological strain variability and include these results in the predictions.

This document specifies a method using automated Randall extraction for the determination of the hexane extract (or light petroleum extract), called the “oil content”, of oilseeds used as industrial raw materials.
The procedure for sunflower seed is different from those for others seeds as it includes an additional moisture content determination after the seed has been ground to prepare the test sample.
If required, the pure seeds and the impurities can be analysed separately. In the case of groundnuts, the pure seeds, the total fines, the non-oleaginous impurities and the oleaginous impurities can be analysed separately.

This document specifies a method for the quantification of individual and/or all fatty acids content in the profile of milk, milk products, infant formula and adult nutritional formula, containing milk fat and/or vegetable oils, supplemented or not supplemented with oils rich in long chain polyunsaturated fatty acids (LC-PUFA). This also includes groups of fatty acids often labelled [i.e. trans fatty acids (TFA), saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), omega-3, omega-6 and omega-9 fatty acids] and/or individual fatty acids [i.e. linoleic acid (LA), α-linolenic acid (ALA), arachidonic acid (ARA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)].
The determination is performed by direct transesterification in food matrices, without prior fat extraction, and consequently it is applicable to liquid samples or reconstituted powder samples with water having total fat ≥ 1,5 % (mass fraction).
The fat extracted from products containing less than 1,5 % (mass fraction) fat can be analysed with the same method after a preliminary fat extraction using methods referenced in Clause 2. Dairy products, such as soft or hard cheeses with acidity level ≤ 1 mmol/100 g of fat, can be analysed after a preliminary fat extraction using methods referenced in Clause 2.
For products supplemented or enriched with PUFA with fish oil or algae origins, the evaporation of solvents is performed at the lowest possible temperature (e.g. max. 40 °C) to recover these sensitive fatty acids.

This document specifies the general requirements and provides guidance for the development and application of loop-mediated isothermal amplification (LAMP) to detect microorganisms and associated genetic markers (e.g. antimicrobial resistance genes, virulence genes) in the food chain.
This document is applicable to all LAMP methods, platforms, and items from the food chain and laboratories.
This document does not apply to the use of LAMP for quantification.
Validation and verification of LAMP methods as either alternative or reference methods are not covered in this document. Both validation and verification of microbiological methods are described in detail in the ISO 16140 series and ISO 17468.
General requirements for isothermal methods including LAMP for molecular biomarker analysis are given in ISO 22942-1, and general requirements and definitions for polymerase chain reaction (PCR) for the detection and quantification of microorganisms in the food chain are given in ISO 22174.
This document has been established for microorganisms in the food chain and 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 for the above items.

This document specifies a method using a farinograph for the determination of the water absorption of flours and the mixing behaviour of doughs made from them by a constant flour mass procedure or by a constant dough mass procedure.
The method is applicable to experimental and commercial flours from wheat (Triticum aestivum L.).
NOTE            This document is related to ICC 115/1[5] and AACC Method 54-21.02[6].

This document specifies methods for the determination of acrylamide in coffee and coffee products by extraction with water, clean-up by solid-phase extraction (SPE) and determination by high-performance liquid chromatography with tandem mass spectrometric detection (HPLC-MS/MS) and gas chromatography with mass spectrometric detection (GC-MS) after derivatization. The methods were validated in a validation study for roasted coffee, soluble coffee, coffee substitutes and coffee products with ranges from 53 μg/kg to 612,1 μg/kg.

This document specifies a procedure for the analysis of residues of highly polar pesticides and metabolites, which are not amenable to common multiresidue methods, in various food commodities of plant and animal origin, including fruits, vegetables, cereals, pulses, oily seeds, nuts, milk, liver and honey. The method was developed at the EURL-SRM hosted at CVUA Stuttgart [1], [2], [3] and has been collaboratively studied on a large number of commodity/pesticide combinations. Guidelines for calibration are outlined in CEN/TS 17061:2019.

1.1 This document specifies four rapid methods for estimating the degree of, or detecting the presence of, hidden insect infestation with sampling (see Clauses 4 to 6) or without sampling (on-site probing, see Clause 7) in a cereal or pulse lot.
NOTE The characteristics leading to the choice of rapid method are summarized in ISO 6639-1:2025, Table 1.
1.2 The method by determination of carbon dioxide production (see Clause 4) is applicable to testing whole grains. It does not apply to testing:
a) finely ground grain products, as there is a risk that particles of material will be sucked up with air samples; or
b) grain products with moisture contents greater than a mass fraction of 15 %, because of the risk of carbon dioxide produced by the products themselves and by microorganisms interfering with the results.
In addition, the method does not apply to the rapid testing of grain products onto which carbon dioxide has already been adsorbed in large quantities (e.g. grain stored in a confined atmosphere, when there are clear external indications of heavy infestation).
The method is applicable to coarsely milled or kibbled grain products, provided that they have been sieved before testing to remove fine particles and loose insects.
The method does not permit the presence of dead adults.
1.3 The whole grain flotation method (see Clause 5) is applicable to detecting hidden infestation in most cereals and pulses but only on a qualitative basis.
1.4 The acoustic method operating on a grain sample (see Clause 6) is applicable to detecting living (and active) insects (larvae and adults before their emergence from a kernel) inside grains in a sample. It does not permit dead adults and larvae or living eggs and pupae (non-feeding stages) to be detected.
1.5 The on-site assessment acoustic method (see Clause 7) is applicable to detecting adult insects and larvae feeding on the grain inside without taking samples. It does not apply to detecting inactive stages (eggs, nymph and moulting).

This document specifies requirements for dried saffron obtained from the pistils of Crocus sativus L. flowers.
It is applicable to saffron in filaments, cut filaments and powder forms.
Recommendations relating to storage and transport conditions are given in Annex A.

This document establishes general principles for the methods of the determination of hidden insect infestation in cereals and pulses.

This document specifies methods of sampling cereals and pulses, in bags or in bulk, for the determination of hidden insect infestation.
This document is applicable to grain in any form of storage structure or transit vehicle at any level of trade from producer to consumer.

This document covers instrumental analysis by elemental analyser-isotope ratio mass spectrometry (EA-IRMS) of food materials to determine C and/or N isotope ratios.
The isotope ratios obtained by following this document are expressed as δ13C and/or δ15N values relative to international measurement standards.
Sample preparation is not included within this document. It is assumed that the food sample has been pre-treated as necessary and homogenized.
Similarly, the interpretation of the obtained isotope delta values is not covered by this document. Following this protocol will result only in isotope delta values for the sample materials.
Solid and/or liquid sample materials can be analysed following this document.
Although other instrumental techniques can be applied to determine δ13C and/or δ15N values in food materials, these other techniques are not covered by this document.

This document specifies a real-time PCR procedure for the quantitation of the amount of roe deer DNA relative to total mammalian DNA in meat and meat products.
Results of this roe deer assay are expressed in terms of roe deer (Capreolus capreolus) haploid genome copy numbers relative to total mammalian haploid genome copy numbers. The content of roe deer can also be expressed as mass fraction in % using gravimetrically prepared calibration material from meat mixtures or model samples.
The method has been previously validated in a collaborative study and applied to DNA extracted from samples that consist of raw roe deer meat in a raw pig meat background as well as raw and boiled sausages.
The limit of detection of the roe deer PCR has been determined experimentally to be at least 5 target gene copies or at least 0,1 % roe deer.
The compliance assessment process is not part of this document.

This document specifies minimum requirements and minimum performance criteria for conducting a single-laboratory validation study for qualitative (binary) real-time polymerase chain reaction (PCR) methods applied to the detection of specific DNA sequences present in foods.
The document is applicable to any single-laboratory validation of a qualitative real-time PCR method used for the detection of specific DNA sequences in food and food products (e.g. for the detection of genetically modified foodstuffs and for species determination, including species known to produce allergenic proteins).
The document does not apply to single laboratory validation of qualitative microbiological real-time PCR methods.
The document does not apply to the evaluation of applicability and practicability with respect to the specific scope of the PCR method.

This document specifies requirements for dried ginger (Zingiber officinale Roscoe) in whole/pieces and ground forms.
Annex A specifies a method for the determination of calcium.
Recommendations for storage and transport conditions are given in Annex B.

This document specifies a method for the determination of total mercury in foodstuffs by cold vapour atomic fluorescence spectrometry (AFS) after pressure digestion.
This method was tested in an interlaboratory study carried out in connection with the pressure digestion method EN 13805 on seven different materials with a mercury concentration in the range from 0,006 mg/kg to 5,38 mg/kg and successfully validated in this range.
The following foodstuffs were analysed:
—   Saithe (dried);
—   Celery (dried);
—   Wheat noodle powder;
—   Wild mushrooms (dried);
—   Pig liver (dried);
—   Cacao powder;
—   Tuna fish (dried).
The lower limit of the method’s applicability varies depending on the food matrix and the water content of the foodstuff. It is a laboratory-specific value and is defined by the laboratory when calculating the limit of quantification (see 9.2).

This document specifies a method for the determination of total mercury in foodstuffs by cold vapour atomic absorption spectrometry (AAS) after pressure digestion.
This method was tested in an interlaboratory study carried out in connection with the pressure digestion method EN 13805 on seven different materials with a mercury concentration in the range from 0,005 mg/kg to 5,06 mg/kg and successfully validated in the range from 0,015 mg/kg to 5,06 mg/kg.
The following foodstuffs were analysed:
—   Saithe (dried);
—   Celery (dried);
—   Wheat noodle powder;
—   Wild mushrooms (dried);
—   Pig liver (dried);
—   Cacao powder;
—   Tuna fish (dried).
The lower limit of the method’s applicability varies depending on the food matrix and the water content of the foodstuff. It is a laboratory-specific value and is defined by the laboratory when calculating the limit of quantification (see 9.2).

This document specifies a method for the determination of total mercury (Hg) in foodstuffs using direct atomic absorption spectrometry after thermal decomposition in an oxygen or air flow and concentration by amalgam formation. The method is applicable for solid and liquid samples.
This method was tested in a interlaboratory study carried out on seven different materials with a mercury concentration in the range from 0,005 mg/kg to 5,20 mg/kg and successfully validated in this range.
The following foodstuffs were analysed:
—   Saithe (dried);
—   Celery (dried);
—   Wheat noodle powder;
—   Wild mushrooms (dried);
—   Pig liver (dried);
—   Cacao powder;
—   Tuna fish (dried).
The lower limit of the method’s applicability varies depending on the food matrix and the water content of the foodstuff. It is a laboratory-specific value and is defined by the laboratory when calculating the limit of quantification (see 9.2).

This document specifies a horizontal in vitro method for the molecular identification and differentiation of the monophasic variant of Salmonella enterica subsp. enterica serovar Typhimurium (1,4,[5],12:i:-) lacking the second H phase H:1,2, starting from isolates. The method detects specific DNA sequences of an intergenic region of the first H phase flagellin cluster for identification of Salmonella enterica subsp. enterica serovar Typhimurium (further called Salmonella Typhimurium) and specific DNA sequences of genes associated with second H phase flagellar antigen expression.
The method is applicable for:
—     differentiation of the isolate under analysis between monophasic Salmonella Typhimurium and the monophasic variant of another Salmonella non-Typhimurium serovar that has the same antigenic formula;
—     identification of the isolate under analysis being either monophasic Salmonella Typhimurium or (biphasic) Salmonella Typhimurium.
This document is applicable for the analysis of a pure culture belonging to the genus Salmonella, isolated from:
—     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 document can also be applied in other domains for identification of monophasic Salmonella Typhimurium (e.g. environmental, human health, animal health).
NOTE            This method has been validated in a method evaluation study and in an interlaboratory study with a large set of different strains (target and non-target strains), isolated from different sources (food products, animals, animal feed, primary production samples and humans). For detailed information on the validation, see Annex E.

This document specifies an amperometric method to determine the content of damaged starch in flour.
It is applicable to all flour samples from the industrial or laboratory milling of wheat (Triticum aestivum L.).
NOTE 1        Wheat can be milled in the laboratory in accordance with the methods described in ISO 27971[9] or in the BIPEA guidance document BY.102.D[10].
NOTE 2        In the absence of validity studies, the results on semi-wholemeal or wholemeal flour, although able to meet the conditions of repeatability given in Clause 9, require careful interpretation.

This document specifies a method using an extensograph for the determination of the rheological properties of wheat flour doughs in an extension test. The recorded load–extension curve is used to assess the general quality of flour and its response to improving agents.
The method is applicable to experimental and commercial flours from wheat (Triticum aestivum L.).
NOTE 1        This document is related to ICC 114[5] and AACC Method 54-10[6].
NOTE 2        For dough preparation, a farinograph is used (see 6.2)

This document specifies a procedure for the parallel determination of glycidol together with 2-MCPD and 3-MCPD present in bound or free form in oils and fats. The method is based on alkaline-catalysed ester cleavage, transformation of the released glycidol into monobromopropanediol (MBPD) and derivatisation of the derived free diols (MCPD and MBPD) with phenylboronic acid (PBA). Though free MCPD and glycidol are supposed to be present in fats and oils in low to negligible quantities only, in the event that free analytes are present, they would contribute proportionately to the results. The results always being the sum of the free and the bound form of a single analyte.
This method is applicable to solid and liquid fats and oils. This document can also apply to animal fats and used frying oils and fats, but a validation study is undertaken before the analysis of these matrices.
Milk and milk products (or fat coming from milk and milk products) are excluded from the scope of this document.

This document specifies a reference method for the determination of the iodine value (commonly known in the industry as IV) of animal and vegetable fats and oils, hereinafter referred to as fats.
Annex B describes a method for the calculation of the IV from fatty acid compositional data. This method is not applicable to fish oils. Furthermore, cold-pressed, crude and unrefined vegetable oils as well as (partially) hydrogenated oils can give different results by the two methods. The calculated IV is affected by impurities and thermal degradation products.
NOTE            The method in Annex B is based upon the AOCS Official method Cd 1c-85[10].

This document specifies a real-time PCR procedure for the quantitation of the amount of equine DNA relative to total mammalian DNA in a raw meat sample.
Results of this equine assay are expressed in terms of equine (Equus genus) haploid genome copy numbers relative to total mammalian haploid genome copy numbers. This assay is specific for representatives of the genus Equus and therefore detects horse, mule, donkey and zebra DNA.
The method has been previously validated in a collaborative study and applied to DNA extracted from samples that consist of raw horse meat in a raw beef (meat) background.
The limit of detection has been determined experimentally to be at least 17 horse haploid genome equivalents (HGE) for both the equine PCR and the mammalian PCR based on the lowest dilution on the respective calibration curves through single laboratory validation. The lowest relative horse content of the target sequence included in the collaborative study was a mass fraction of 0,1 % based on gravimetrically prepared raw horse muscle tissue in a raw beef muscle tissue background.
The compliance assessment process is not part of this document.

This document specifies a method for the determination of the ratio of stable isotopes of carbon (13C/12C) of sugars contained in honey by using liquid chromatography coupled to an isotope ratio mass spectrometer (LC-IRMS) for compound separation and subsequent determination of the 13C/12C ratio of mono-, di-, and trisaccharides. These ratios can be used to assess honey authenticity by comparing them to guidance values of genuine honey, which have been previously agreed by subject matter experts, as the 13C/12C ratios of sugars of genuine honey and sugars contained in adulterants (syrups made from starch-rich plants or from sugar cane or sugar beet) differ to a certain extent.
The compliance assessment process is not part of this document.

This document specifies the general principle and the technical protocol for the validation of identification methods of microorganisms for microbiology in the food chain. As there is no reference method, no method comparison study can be run. Therefore, this document provides a protocol to evaluate the performance characteristics and validate the method workflow using well-defined strains. When required, an additional identification method can be used.
This document is applicable to the validation of identification methods of microorganisms that are used for the analysis of isolated colonies from:
—     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.
Identification methods only validated in accordance with this document cannot be used instead of confirmation described in:
—     the reference method;
—     an alternative method validated in accordance with ISO 16140-2;
—     an alternative method validated in accordance with ISO 16140-6.
In these instances, the identification method is validated in accordance with ISO 16140-6 method that is used as a confirmation method.
This document is applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms, which can be determined on a case-by-case basis.

This document specifies an isotopic method to control the authenticity of vinegar and food containing vinegar as an ingredient (for example Aceto Balsamico di Modena), with a density below 1,28 g/cm3. This method is applicable on acetic acid of vinegar (from wine, cider, agricultural alcohol, etc.) in order to characterize the botanical origin of acetic acid and to detect adulterations of vinegar using synthetic acetic acid or acetic acid from a non-allowed origin (together with the method described in EN 16466 2).
The isotopic analysis of the extracted acetic acid by 2H-NMR is based on a similar method already normalized for wine analysis[10].

This document specifies a method for the determination of soluble 16-O-Methylcafestol and 16-O-Methylkahweol content (the sum of free forms and derivatives, e.g. fatty acid esters, henceforth abbreviated as 16-OMD = “diterpenes”) in roasted coffee (beans or ground), using quantitative proton nuclear magnetic resonance spectroscopy (1H-qNMR).
If complying with the experimental parameters described below, this test procedure has been proven for the following mass fraction range:
w16-OMD:    20 mg/kg to 2 000 mg/kg.
The mass fraction range can be expanded by suitable changes of the experimental parameters, e.g. a different weighed portion of ground coffee or the accumulation of more NMR-transients.

This document specifies a high-performance liquid chromatography (HPLC) method for determining the 16-O-Methylcafestol content in green and roasted coffee.
The method is suitable for a content of 40 mg/kg to 1 600 mg/kg of 16-O-Methylcafestol of green and roasted coffee, respectively. The collaborative study has shown that mass fractions also between 20 mg/kg to 40 mg/kg can be successfully analysed depending on the laboratory equipment.
The compliance assessment process is not part of this document.

This document provides technical definitions of terms relating to authenticity and fraud when referring to food products. All terms and definitions are in the context of food supply chains, but most of them can also be applied when referring to feed products and the feed supply chain.

This document specifies a method to determine the free glycerol and residual mono-, di- and triglyceride contents in fatty acid methyl esters (FAME). The total glycerol content is then calculated from the obtained results.
Under the conditions described, the quantification limits are 0,001 % (m/m) for free glycerol, 0,10 % (m/m) for all glycerides (mono-, di- and tri-). This method is suitable for FAME prepared from rapeseed, sunflower, soybean, palm, animal oils and fats and mixture of them. It is not suitable for FAME produced from or containing coconut and palm kernel oils derivatives because of overlapping of different glyceride peaks.
NOTE 1   For the purposes of this document, the term "% (m/m)" is used to represent the mass fraction.
NOTE 2   Under the common EN 14105 GC conditions, squalene can coelute with alpha glycerol monostearate. If the presence of squalene is suspected, EN 17057 can be used to discriminate between squalene and glycerol monostearate.
WARNING - The use of this document can involve hazardous materials, operations and equipment. This document does not purport to address all of the safety problems associated with its use. It is the responsibility of users of this document to take appropriate measures to ensure the safety and health of personnel prior to application of the standard, and fulfil statutory and regulatory requirements for this purpose.

This document specifies a routine reference method for the determination of the moisture content of cereals and cereal products.
This document applies to: wheat, rice (paddy, husked and milled), barley, millet (Panicum miliaceum), rye, oats, triticale, sorghum in the form of grains, milled grains, semolina or flour.
The method is not applicable to maize and pulses.
NOTE            For moisture content determination in maize, see ISO 6540[5]; and for pulses, see ISO 24557[7].

This document specifies an automatic method for the reference method (see ISO 712-1) for the determination of moisture content of cereals and cereal products using an automatic drying oven.
This document is applicable to wheat, rice (paddy, husked and milled), barley, millet (Panicum miliaceum), rye, oats, triticale, sorghum in the form of grains, milled grains, semolina and flour.
The method does not apply to maize and pulses.
NOTE            For moisture content determination in maize, see ISO 6540, and for pulses, see ISO 24557.

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.