This document specifies a procedure for the detection of a DNA sequence present in a genetically modified linseed (Linum usitatissimum) line (event FP967, also named as “CDC Triffid”). For this purpose, extracted DNA is used in a real-time PCR and the genetic modification (GM) is specifically detected by amplification of a 105 bp DNA sequence representing the transition between the nopaline synthase gene terminator (Tnos) from Agrobacterium tumefaciens and the dihydrofolate reductase gene (dfrA1) from a Class 1 integron of Escherichia coli. The method described 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 and seeds. The application of this method requires the extraction of an adequate amount of amplifiable DNA from the relevant matrix for the purpose of analysis.

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This document specifies a method for the semi-quantitative analysis of oils, fats and oil/fat-related samples (deodistillates). It is applicable to the screening of oils, fats and oil/fat-related samples to obtain main (e.g. triglycerides) and minor (e.g. sterols, sterol esters, tocopherols, wax esters, fatty alcohols, glycerol) component information in one single analysis. For a truly quantitative analysis of pre-identified compound classes, specific methods are more appropriate. The method can also be used as a useful qualitative screening tool for the relative comparison of sample compositions.

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This document specifies a high performance liquid chromatography (HPLC) or ultra-high performance liquid chromatography (UHPLC) method for the determination of content of the four major theaflavins of tea. It is applicable to both leaf and instant black and oolong teas. The method is currently not validated for ready-to-drink (RTD) beverages.

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This document specifies two methods for the determination of the melting point in open capillary tubes, commonly known as the slip melting point, of animal and vegetable fats and oils (referred to as fats hereinafter). —   Method A is only applicable to animal and vegetable fats which are solid at ambient temperature and which do not exhibit pronounced polymorphism. —   Method B is applicable to all animal and vegetable fats which are solid at ambient temperature and is the method to be used for fats whose polymorphic behaviour is unknown. For the determination of the slip melting point of palm oil samples the method given in Annex A shall be used. NOTE 1  If applied to fats with pronounced polymorphism, method A will give different and less satisfactory results than method B. NOTE 2  Fats which exhibit pronounced polymorphism are principally cocoa butter and fats containing appreciable quantities of 2-unsaturated, 1,3-saturated triacylglycerols.

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This document establishes the minimum specifications for rice (Oryza sativa L.) that is subject to international trade. It is applicable to husked rice and milled rice (aromatic and not aromatic), parboiled or not, intended for direct human consumption. It does not apply to other products derived from rice nor to waxy rice (glutinous rice).

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This document specifies a method for applying magnitude estimation to the evaluation of sensory attributes. The methodology specified covers the training of assessors, and obtaining magnitude estimations as well as their statistical interpretation.

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This document defines terms for classifying and assessing green tea for commerce.

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This document provides guidelines on: — how to express vitamin quantity, — the expression of different molecular forms in appropriate units, — and in some cases, vitamin activity, according to vitamers present or used in food products, in addition to the quantitative content determination obtained from ISO and CEN analytical standards. It provides information to be used as a basis for discussion between stakeholders and food control laboratories. It is not intended to be prescriptive or exhaustive.

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This document specifies a method for the determination of aflatoxin M1 content in milk and milk powder. The lowest level of validation is 0,08 µg/kg for whole milk powder, i.e. 0,008 µg/l for reconstituted liquid milk. The limit of detection (LOD) is 0,05 μg/kg for milk powder and 0,005 μg/kg for liquid milk. The limit of quantification (LOQ) is 0,1 μg/kg for milk powder and 0,01 μg/kg for liquid milk. The method is also applicable to low-fat milk, skimmed milk, low-fat milk powder and skimmed milk powder.

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This document specifies a method for the detection of Anisakidae L3 larvae commonly found in marine and anadromous fishes. The method is applicable to fresh fish and/or frozen fish, as well as lightly processed fish products, such as marinated, salted or cold smoked. This method is applicable to quantifying parasitic infections by estimating the number of parasites in the fish musculature. This method does not apply to determining the species or genotype of detected parasites. Final identification is made by morphological and/or molecular methods.

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This document specifies a method for the detection of Anisakidae L3 larvae commonly found in marine and anadromous fishes. The method is applicable to fresh fish and/or frozen fish, as well as lightly processed fish products, such as marinated, salted or smoked. It is also suitable for visceral organs as a confirmatory method for a visual inspection scheme. The artificial digestion method[4][5][6] is applicable to quantifying parasitic infections by estimating the number of parasites in the fish musculature and, when applied to fresh fish or lightly processed fish products (never frozen before processing), determining the viability of Anisakidae L3, which can be present. This method does not apply to determining the species or genotype of detected parasites. Final identification is made by morphological and/or molecular methods.

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This document specifies a method for the determination of the moisture and volatile matter content of oilseed meals obtained by the extraction of oil from oilseeds by pressure and/or solvent.

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This document specifies requirements for the determination of the dry matter content of liquid or pasty coffee extracts by means of the sea sand method. It is applicable to liquid or pasty coffee extracts. The method is intended as a routine method.

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This document gives guidelines for substantiating sensory claims on food and non-food products and their packaging for advertising consumer-packaged goods. This document differentiates sensory claims from other types of claims. It provides classification and examples of the different types of sensory claims. It highlights special issues associated with testing to substantiate sensory claims. It includes case studies and references. This document does not apply to: — specific or detailed requirements for different test methods that are used to support sensory claims; — factual claims regarding a product's country of origin, ingredients, processing and nutritional components; — factual claims regarding the technical features of the product; — claims regarding a product's health, medical or therapeutic benefits, physiological effects, structure or function benefits when consumed or applied to the human body; — claims based on instrumental assessments of the attributes or performance of a product (i.e. instrumental assessments; in this case, test methods are used in which no human participant evaluates the product and/or no human participant provides a response to a product); — claims about services (e.g. a house cleaning service, airline services, automobile services); — claims about large/slow moving consumer goods (autos, refrigerators, stoves, etc.).

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The document specifies the definitions and technical criteria to be fulfilled for foods and food ingredients to be suitable for vegetarians (including ovo-lacto-, ovo- and lacto-vegetarians) or vegans as well as for food labelling and claims. It is applicable to business-to-business communication (B2B), to the food trade, and to food labelling and claims. The definitions and technical criteria apply only post-harvest/collecting. It does not apply to human safety, environmental safety, socio-economic considerations (e.g. fair trade, animal welfare), religious beliefs and the characteristics of packaging materials.

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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 more efficient than the duo-trio test (described in ISO 10399), but has limited use with products that exhibit strong carryover and/or lingering flavours. 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 homogeneous. The method is effective for: a) determining that: either a perceptible difference results (triangle testing for difference); a perceptible difference does not result (triangle testing for similarity), when, for example, a change is made in ingredients, processing, packaging, handling or storage; b) selecting, training and monitoring assessors.

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This document gives guidelines for the establishment of a conversion relationship between the results of an alternative method and an anchor method, and its verification for the quantitative determination of the microbiological quality of milk. NOTE The conversion relationship can be used a) to convert results from an alternative method to the anchor basis or b) to convert results/limits, expressed on an anchor basis, to results in units of an alternative method.

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This document specifies two methods: - a reference method for the determination of the moisture content of maize grains and ground whole maize, groats, grits and maize flour, see Clause 4; - a routine method for the evaluation of the moisture content of maize in whole grains, see Clause 5. The latter is not suitable for use for experts' reports, or for calibration or checking of humidity meters, because of its significant bias to the reference method (see Table B.3).

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This document specifies requirements and test methods for the dried barberry fruit of the Berberis vulgaris L. tree. It is applicable to dried red barberries only.

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This document specifies the quantitative liquid chromatographic determination of specific sugars (galactose, glucose, fructose, sucrose, lactose and maltose) in various milk and milk products, applying arabinose as an internal standard. The method is applicable to the following dairy matrices: milk, sweetened condensed milk, milk powder, cheese, whey powder, infant formula, milk dessert and yoghurt. The method does not apply to dairy products containing soy or to the determination of the lactose content in low-lactose milk products at levels below 1 mg/g. A high performance anion exchange chromatography method in combination with pulsed amperometric detection (HPAEC-PAD) method is applied[5][3][4]. With this method, thirteen different monosaccharides, disaccharides and trisaccharides can be separated: fucose, arabinose, galactose, glucose, fructose, sucrose, lactose, lactulose, maltose, melibiose, trehalose, isomaltulose and maltotriose. The method is applicable to labelling for the six most important sugars that can be present by nature or by addition in milk and milk products. The method does not apply to sugar contents less than 0,1 %.

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This document specifies requirements and methods of sampling and testing for ground cassava leaves, which are obtained from the processing of fresh cassava leaves (Manihot esculenta Crantz or Manihot glaziovii) intended for human consumption.

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This document specifies the protocol for the verification of reference methods and validated alternative methods for implementation in the user laboratory. This document is applicable to the verification of methods used for the analysis (detection and/or quantification), confirmation and typing of 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. This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms or their metabolites, to be determined on a case-by-case basis. The technical protocols for the verification of validated qualitative methods and validated quantitative methods are described in Clauses 5 and 6. The technical protocol for the verification of validated alternative confirmation and typing methods is described in Clause 7. The protocols for the verification of non-validated reference methods are described in Annex F.

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This document establishes minimum specifications for wheat (Triticum aestivum L.) grains intended for human consumption and which are the subject of international trade. It is also applicable to local wheat trade. NOTE Wheat (Triticum aestivum L.) is also called "common wheat" in some regions.

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This document specifies an entrainment method for the determination of the moisture content of spices and condiments.

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This document specifies the categories of pao cai (salted fermented vegetables) and its requirements, including sensory, physical and chemical, safety, labelling, transport and storage. It also describes the corresponding test methods. This document does not apply to kimchi.

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This document specifies a method for the simultaneous quantitative determination of four water-soluble vitamins in infant formula and related nutritional products, including relevant forms of vitamins B1, B2, B3 and B6 by enzymatic digestion and UHPLC-MS/MS. This document is not intended to be used on products where vitamins have not been added.

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This document specifies a method for the direct enumeration of potentially enteropathogenic V. parahaemolyticus (tdh and/or trh positive) and/or the enumeration of total V. parahaemolyticus in seafood.

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This document gives guidelines for the use of near infrared (NIR) spectrometry in the analysis of milk and milk products in liquid, semi-solid or solid form. Depending on the sample form and application, different instrument setups for transmittance, diffuse reflectance or transflectance can be applied.

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This document specifies a method for the determination of total or free choline and carnitine in infant formula and adult nutritionals by liquid chromatography and tandem mass spectrometry (HPLC-MS/MS).

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This document specifies a liquid chromatography tandem mass spectrometry (LC?MS/MS) method for the quantification of the inhibitory substance, nitrofurazone, in milk and milk products. The method has been validated for measuring trace levels of intact nitrofurazone to levels down to 1 ng/g in fluid milk and powdered dairy products on a whole product (i.e. powder) basis. While the method is expected to apply to other dairy matrices, additional validation will be required to demonstrate this.

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This document specifies a method for the determination of inulin-type fructans (including oligofructose, fructooligosaccharides) in infant formula and adult nutritionals (both powder and liquid) containing 0,03 g/100 g to 5,0 g/100 g of fructans in the product as prepared ready for consumption. The method has been validated in a multi laboratory study[1] with reconstituted standard reference material (SRM), infant/adult nutritional formula at a level of 0,204 g/100 g, adult nutritionals ready-to-feed (RTF) at levels of 1,28 g/100 g and 2,67 g/100 g, infant formula RTF at a level of 0,300 g/100 g, reconstituted follow-up formula at levels of 0,209 g/100 g to 0,275 g/100 g, reconstituted infant formula at levels from 0,030 8 g/100 g to 0,264 g/100 g. During the single laboratory validation study[2], spike-recovery experiments were performed up to 5 g/100 g in reconstituted infant formula powders (milk-based, partially hydrolysed milk-based and soy-based), adult nutritional RTF and reconstituted adult nutritional powders.

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This document specifies requirements and test methods for fresh artichokes, including their hearts and bottoms, of the following groups: — cynara cardunculus Scolymus Group; — cynara cardunculus Cardoon Group, syn. C. cardunculus var. altilis DC. It does not apply to processed artichokes.

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This document specifies requirements and test methods for fresh asparagus shoots of commercial varieties of asparagus grown from Asparagus officinalis L., of the Liliaceae family, offered to consumers after preparation and packaging. This document is applicable to all asparagus except green and violet/green asparagus with a diameter less than 3 mm and white and violet asparagus with a diameter less than 8 mm, packed in uniform bundles or unit packages. This document does not apply to processed asparagus.

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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of porcine-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of porcine material derived from pig (Sus scrofa domesticus) and wild boar (Sus scrofa). The target sequence is a partial fragment of the Sus scrofa beta actin (ACTB) gene, partial cds. (i.e. GenBank accession number DQ452569.1)[1], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of bovine-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of bovine material derived from cattle including taurine (Bos taurus) and zebu (Bos indicus). The assay also detects the species bison (Bison bison) and yak (Bos mutus). The target sequence is a partial fragment of the bovine nuclear beta actin gene in chromosome 25 (i.e. GenBank accession number NC_037352.1)[1][2][3], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of goat-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of goat material derived from goat (Capra hircus). The target sequence is a partial fragment of the goat chromosome 9 DNA sequence (i.e. GenBank accession number NC_030816.1)[1], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of donkey-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of donkey material derived from donkey (Equus asinus), mule (Equus caballus ♀ × Equus asinus ♂) and hinny (Equus caballus ♂ × Equus asinus ♀). The assay also detects the species zebra (Equus burchellii). The target sequence is a partial fragment of the Equus asinus isolate Maral har breed Guanzhong donkey unplaced genomic scaffold, ASM130575v1 scaffold786, whole genome shotgun sequence (i.e. GenBank accession number NW_014638576.1)[1], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of ovine-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of ovine material derived from sheep (Ovis aries). The target sequence is a partial fragment of the ovine nuclear prolactin receptor short form mRNA gene (PRLR) (i.e. GenBank accession number AF041979.1)[1], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of chicken-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of chicken material derived from chicken (Gallus gallus domesticus) and jungle fowl (Gallus gallus). The target sequence is a partial fragment of the Gallus gallus transforming growth factor beta 3, intron 4 (TGF-β3) gene (i.e. GenBank accession number AY685072.1)[1], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

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This document specifies a real-time polymerase chain reaction (real-time PCR) method for the qualitative detection of horse-specific DNA derived from food and feed. It requires the extraction of an adequate amount of PCR amplifiable DNA from the relevant matrix and can be applied to the detection of horse material derived from domestic horse (Equus caballus), mule (Equus caballus ♀ × Equus asinus ♂), hinny (Equus caballus ♂ × Equus asinus ♀) and zebroid (Equus caballus × Equus simplicidens). The assay also detects the species Przewalski's horse (Equus przewalskii) and zebra (Equus burchellii). The target sequence is an Equus caballus isolate Twilight breed thoroughbred chromosome 28, EquCab3.0, whole genome shotgun sequence (i.e. GenBank accession number NC_009171.3)[1], which is present as a single copy per haploid genome. The provided PCR assay for this target has an absolute limit of detection of five copies per reaction, with ≥ 95 % replicability at this concentration (LOD95 %).

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This document specifies a test method to determine the extractable colour in paprika by measuring the absorbance of an acetone extract of the sample. It is applicable to ground paprika in every presentation (sweet, hot, smoked, etc).

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This document specifies requirements for ground sweet and hot paprika (Capsicum annuum L. and Capsicum frutescens L.). Recommendations relating to storage and transport conditions are given in Annex A. A list of terms used in different countries for paprika is given in Annex B. This document does not apply to ground chillies and other species of capsicums. NOTE Specifications for ground chillies and capsicums are given in ISO 972.

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This document specifies a method for the quantitative determination of hydroxytyrosol and tyrosol content in extra virgin olive oils using reverse phase high performance liquid chromatography (RP-HPLC) with spectrophotometric detection. The method is also applicable to all other olive oils of a different commercial category.

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This document specifies a reference method for the determination of the amylose content of milled rice, non-parboiled. The method is applicable to rice with an amylose mass fraction higher than 5 %. This document can also be used for husked rice, maize, millet and other cereals if the extension of this scope has been validated by the user. NOTE Amylose values determined with this document can be compared with PDO and PGI legislation.

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This document specifies two simplified routine methods for the determination of the amylose mass fraction of milled rice, non-parboiled. The main difference between the two methods is the dispersion procedure: method A specifies hot dispersion, and method B specifies cold dispersion. Both methods are applicable to rice with an amylose mass fraction higher than 5 %. NOTE These methods describe simplified procedures for the preparation of samples, which are frequently used in routine laboratories. The methods use the same reagents as the reference method (see ISO 6647-1), but omit the defatting step. Rice samples where the amylose mass fraction has been determined by the reference method are used as standards.

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This document specifies the general principles and the technical protocols (based on orthogonal, factorial studies) for the validation of non-proprietary methods for microbiology of the food chain. This document is applicable to the validation of methods used for the analysis (detection or quantification) of 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. This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms or their metabolites, to be determined on a case-by-case basis. This document specifies protocols for the validation against a reference method for both quantitative and qualitative methods. This document also provides a protocol for the validation of quantitative methods without a reference method. Qualitative methods cannot be validated without a reference method in accordance with this document. NOTE ISO 16140-2 specifies the general principle and the technical protocol for the validation of alternative, mostly proprietary, methods against a reference method. This document is only applicable to the validation of methods that are fully specified with regard to all relevant parameters (including tolerances on temperatures and specifications on culture media) and that have already been optimized. Methods that have been validated in accordance with this document can be used by the laboratories of the specified population of laboratories.

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This document specifies the general principles and the technical protocols for single-laboratory validation of methods for microbiology in the food chain. The protocols in this document only validate the method for the laboratory conducting the study. This document is applicable to single-laboratory validation of: — methods used in the analysis (detection or quantification) of 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; — methods for the confirmation or typing of microorganisms. This validation will replace only the confirmation or typing procedure of a specified method (see Annex G). This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms or their metabolites, to be determined on a case-by-case basis. Single-laboratory validation is required if an interlaboratory validation in accordance with ISO 16140-2 is not appropriate. Possible applications are: — validation of an in-house method; — method evaluation study in the validation process of a reference method in accordance with ISO 17468; — extension of the scope of an ISO 16140-2 validated method, e.g. category extension or test portion size; — modifications of existing methods. Single-laboratory validation is the second step in the standardization of a reference method (see ISO 17468). It is only applicable to methods that are fully specified with regard to all relevant parameters (including tolerances on temperatures and specifications on culture media) and that have already been optimized.

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This document specifies a method for the quantitative determination of β-carotene and lycopene in infant formula and adult nutritionals in solid (i.e. powders) or liquid (i.e. ready-to-feed liquids and liquid concentrates) forms using reversed-phase ultra-high performance liquid chromatography (RP-UHPLC) and UV-visible detection. The application range runs from 1 μg/100 g to 1 500 μg/100 g for lycopene and from 1 μg/100 g to 2 250 μg/100 g for β-carotene. Based on the single-laboratory validation, the limit of detection (LOD) was 0,1 μg/100 g and the limit of quantification (LOQ) was 0,3 μg/100 g for each carotenoid. The method does not apply to materials that contain measurable levels of β-apo-8′-carotenal. The reproducibility data meets the requirements given in References [8] and [10]. Annex C specifies the determination of lutein. The reproducibility data does not meet the requirements given in Reference [9].

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