1、June 2010 Translation by DIN-Sprachendienst.English price group 12No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 6
2、7.050!$i,“1700904www.din.deDDIN EN 15842Foodstuffs Detection of food allergens General considerations and validation of methodsEnglish translation of DIN EN 15842:2010-06Lebensmittel Nachweis von Lebensmittelallergenen Allgemeine Betrachtungen und Validierung von VerfahrenEnglische bersetzung von DI
3、N EN 15842:2010-06Produits alimentaires Dtection des allergnes alimentaires Considrations gnrales et validation des mthodesTraduction anglaise de DIN EN 15842:2010-06www.beuth.deDocument comprises pagesIn case of doubt, the German-language original shall be considered authoritative.2106.10 DIN EN 15
4、842:2010-06 A comma is used as the decimal marker. National foreword This standard has been prepared by Technical Committee CEN/TC 275 “Food analysis Horizontal methods”, Working Group WG 12 “Food allergens” (Secretariat: DIN, Germany). The responsible German body involved in its preparation was the
5、 Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-01-05 AA Lebensmittelallergene. German experts made a significant contribution to the development of this standard. The DIN Standards corresponding to the Int
6、ernational Standards referred to in this document are as follows: ISO 5725-1 DIN ISO 5725-1 ISO 5725-2 DIN ISO 5725-2 ISO 11843-1 DIN ISO 11843-1 National Annex NA (informative) Bibliography DIN ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principl
7、es and definitions DIN ISO 5725-2, Accuracy (trueness and precision) of measurement methods and results Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method DIN ISO 11843-1, Capability of detection Part 1: Terms and definitions 2 EUROPEAN S
8、TANDARD NORME EUROPENNE EUROPISCHE NORM EN 15842 February 2010 ICS 67.050 English Version Foodstuffs - Detection of food allergens - General considerations and validation of methods Produits alimentaires - Dtection des allergnes alimentaires - Considrations gnrales et validation des mthodes Lebensmi
9、ttel - Nachweis von Lebensmittelallergenen - Allgemeine Betrachtungen und Validierung von VerfahrenThis European Standard was approved by CEN on 25 December 2009. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standar
10、d the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN Management Centre or to any CEN member. This European Standard exists in three official versions (English, French,
11、German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cy
12、prus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION
13、COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2010 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 15842:2010: EEN 15842:2010 (E) 2 Contents Page Foreword 3Introd
14、uction .41 Scope 52 Normative references 53 Terms and definitions .54 General aspects for the use of reference materials in food allergen analysis . 124.1 Reference material 124.2 Reference method. 134.3 General requirements for production and storage of reference materials . 135 Guidance to the use
15、r for selection of methods . 145.1 General . 145.2 Immunoassay based methods 145.3 Molecular biology based methods 155.4 Chromatographic methods 156 Laboratory organisation 156.1 General . 156.2 Laboratory design. 157 Procedure 157.1 General . 157.2 Preparation of sample 167.3 Extraction 167.4 Prepa
16、ration of calibration curves 167.5 Assay procedure . 167.6 Quality assurance requirements . 168 Interpretation and expression of the results . 168.1 General . 168.2 Quantitative analysis 168.3 Qualitative analysis 168.4 Provisions 178.5 Ambiguous results . 179 Test report . 17Bibliography . 18DIN EN
17、 15842:2010-06 EN 15842:2010 (E) 3 Foreword This document (EN 15842:2010) has been prepared by Technical Committee CEN/TC 275 “Food Analysis Horizontal Methods”, the secretariat of which is held by DIN. This European Standard shall be given the status of a national standard, either by publication of
18、 an identical text or by endorsement, at the latest by August 2010, and conflicting national standards shall be withdrawn at the latest by August 2010. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not b
19、e held responsible for identifying any or all such patent rights. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Es
20、tonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and the United Kingdom. DIN EN 15842:2010-06 EN 15842:2010 (E) 4 Introduction The main focus
21、 of this European Standard is on immunoassays, chromotographic and nucleic acid based methods for the determination of food allergens. However, because of the rapid developments in this area, other technologies may be considered. The search for food allergens is performed by means of the following s
22、uccessive (or simultaneous) steps. After sample collection, proteins, nucleic acids or other markers are extracted from the test portion. Extracted analytes can be further purified, simultaneously or after the extraction process. Afterwards, they are diluted (if necessary) and subjected to analytica
23、l procedures such as immunoassays (e.g. ELISA), nucleic acid based assays (e.g. PCR) or chromatographic (e.g. LC-MS). These steps are detailed in this document and in the following documents: EN 15633-1:2009, Foodstuffs Detection of food allergens by immunological methods Part 1: General considerati
24、ons EN 15634-1:2009, Foodstuffs Detection of food allergens by molecular biological methods Part 1: General considerations DIN EN 15842:2010-06 EN 15842:2010 (E) 5 1 Scope This European Standard specifies how to use the standards for immunoassays, nucleic based and chromatographic methods and their
25、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 mat
26、erials 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.
27、As new materials become accepted and validated, they may 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. 2 Normative references The following referenced documents
28、 are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. EN ISO/IEC 17025, General requirement for the competence of testing and calibration
29、 laboratories (ISO/IEC 17025:2005) EN ISO 17511:2003, In vitro diagnostic medical devices Measurement of quantities in biological samples Metrological traceability of values assigned to calibrators and control materials (ISO 17511:2003) ISO Guide 31, Reference materials Contents of certificates and
30、labels ISO Guide 35, Reference materials General and statistical principles for certification 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 accepted reference value value that serves as an agreed-upon reference for comparison and which is d
31、erived as: theoretical or established value, based on scientific principles, an assigned value, based on experimental work of some national or international organization, consensus value, based on collaborative experimental work under the auspices of a scientific or engineering group ISO Guide 30:19
32、92 3.2 accuracy closeness of agreement between a test result or measurement result and the true value DIN EN 15842:2010-06 EN 15842:2010 (E) 6 NOTE 1 In practice, the accepted reference value is substituted for the true value. NOTE 2 The term “accuracy“, when applied to a set of test or measurement
33、results, involves a combination of random components and a common systematic error or a bias component. NOTE 3 Accuracy refers to a combination of trueness and precision. ISO 3534-2:2006 3.3 applicability range quantity interval within which the analytical procedure has been demonstrated by collabor
34、ative trial or other appropriate validation to have a suitable level of precision and accuracy EN ISO 24276:2006 3.4 bias difference between the expectation of a test result or measurement result and a true value NOTE 1 Bias is the total systematic error as contrasted to random error. There may be o
35、ne or more systematic error components contributing to the bias. A larger systematic difference from the accepted reference value is reflected by a larger bias value. NOTE 2 The bias of a measuring instrument is normally estimated by averaging the error of indication over an appropriate number of re
36、peated measurements. The error of indication is the “indication of a measuring instrument minus a true value of the corresponding input quantity“. ISO 3534-2:2006 3.5 Certified Reference Material CRM reference material, accompanied by a certificate, one or more of whose property values are certified
37、 by a procedure which establishes its traceability to an accurate realisation of the unit in which the property values are expressed, and for which each certified value is accompanied by an uncertainty at a stated level of confidence ISO Guide 30:1992 3.6 certified value for a CRM, value that appear
38、s in the certificate accompanying the material ISO Guide 30:1992 3.7 characterization for a reference material, determination of one or more physical, chemical, biological, or technological property values that are relevant to its intended end use ISO Guide 30:1992 3.8 collaborative study interlabor
39、atory study interlaboratory study in which each laboratory uses a defined method of analysis to analyse identical portions of homogenous material to assess the performance characteristics obtained for the method of analysis DIN EN 15842:2010-06 EN 15842:2010 (E) 7 NOTE Guidelines for performing coll
40、aborative trials are elaborated in ISO 5725-1 3 and in IUPAC harmonized protocol 1995 16. 3.9 commutability of a material closeness of agreement between the mathematical relationship of the measurement results obtained by two measurement procedures for a stated quantity in a given material, and the
41、mathematical relationship obtained for the quantity in routine samples EN ISO 17511:2003 3.10 consensus value (of a given quantity) for a reference material, value of the quantity obtained by interlaboratory testing, or by agreement between appropriate bodies or experts ISO Guide 30:1992 3.11 fitnes
42、s for purpose applicability degree to which data produced by a measurement process enables a user to make technically and administratively correct decisions for a stated purpose NOTE See 17. 3.12 homogeneity condition of being of uniform structure or composition with respect to one or more specified
43、 properties NOTE A reference material is said to be homogeneous with respect to a specified property if the property value, as determined by tests on samples of specified size, is found to lie within the specified uncertainty limits, the samples being taken either from different supply units (bottle
44、s, packages, etc.) or from a single supply unit. Adapted from ISO Guide 30:1992 3.13 laboratory sample sample as prepared for sending to the laboratory and intended for inspection or testing ISO 78-2:1999 3.14 limit of detection LOD minimum amount or concentration of the analyte in test sample which
45、 can be detected reliably but not necessarily quantified, as demonstrated by a collaborative trial or other appropriate validation 3.15 limit of detection for quantitative determinations amount of an analyte corresponding to the lowest measurement signal which with a closely defined confidence may b
46、e interpreted as indicating that the analyte is present in the sample, but without allowing exact quantification 3.16 limit of detection for qualitative determinations threshold concentration below which positive identification is unreliable according to the established requirements for reliability
47、DIN EN 15842:2010-06 EN 15842:2010 (E) 8 NOTE See 24. 3.17 limit of quantitation LOQ lowest concentration or amount of the analyte in a test sample which can be quantitatively determined with an acceptable level of precision and accuracy, as demonstrated by collaborative trail or other appropriate v
48、alidation NOTE See 24. 3.18 limit of quantification limit of determination lowest amount of an analyte which can be determined quantitatively with a closely defined confidence NOTE See 24. 3.19 linearity ability to elicit test results that are directly, or by means of well defined, mathematical tran
49、sformations, proportional to the concentration of analyte in samples within a given range NOTE See 15. 3.20 matrix all compounds in the sample with the analyte NOTE Each matrix has generally a common name which permits classification. EN ISO 21572:2004 3.21 outlier member of a set of values which is inconsistent with the other members of that set NOTE ISO 5725 specifies the statistical tests and the significance level used to ide
