1、December 2015 English price group 9No 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 67.200.20!%J|“2399689www.din.deD
2、IN EN ISO 10519Rapeseed Determination of chlorophyll content Spectrometric method (ISO 10519:2015);English version EN ISO 10519:2015,English translation of DIN EN ISO 10519:2015-12Rapssamen Bestimmung des Chlorophyllgehaltes Spektrometrisches Verfahren (ISO 10519:2015);Englische Fassung EN ISO 10519
3、:2015,Englische bersetzung von DIN EN ISO 10519:2015-12Graines de colza Dtermination de la teneur en chlorophylle Mthode spectromtrique (ISO 10519:2015);Version anglaise EN ISO 10519:2015,Traduction anglaise de DIN EN ISO 10519:2015-12SupersedesDIN EN ISO 10519:2000-12www.beuth.deDocument comprises
4、DTranslation by DIN-Sprachendienst.In case of doubt, the German-language original shall be considered authoritative.14 pages01.16DIN EN ISO 10519:2015-12 2 A comma is used as the decimal marker. National foreword This document (EN ISO 10519:2015) has been prepared by Technical Committee ISO/TC 34 “F
5、ood products” (Secretariat: AFNOR, France) in collaboration with Technical Committee CEN/TC 307 “Oil seeds, vegetable and animal fats and oils and their by-products Methods of sampling and analysis” (Secretariat: AFNOR, France). The responsible German bodies involved in its preparation were DIN-Norm
6、enausschuss Lebensmittel und landwirtschaftliche Produkte (DIN Standards Committee Food and Agricultural Products) and the Deutsche Gesellschaft fr Fettwissenschaft (German Fat Research Society), Joint Committee NA 057-05-05 AA Gemeinschaftsausschuss fr die Analytik von Fetten, len, Fettprodukten, v
7、erwandten Stoffen und Rohstoffen. The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 542 DIN EN ISO 542 ISO 648 DIN EN ISO 648 ISO 664 DIN EN ISO 664 ISO 665 DIN EN ISO 665 ISO 5725-1 DIN ISO 5725-1 Amendments This standard differs from DI
8、N EN ISO 10519:2000-12 as follows: a) the standards cited in Clause 2 are now undated references; b) the ball mill used for testing (see Subclauses 6.3, 9.1 and 9.2.1) shall now be a commercial ball mill (previously a Dangoumau ball mill was requested); c) footnote 1) to Subclause 6.3 concerning the
9、 Dangoumau ball mill has been deleted; d) Annex ZA (normative) “Normative references to international publications with their relevant European publications” has been deleted; e) the standard has been editorially revised and brought in line with the current rules of presentation. Previous editions D
10、IN EN ISO 10519: 2000-12 DIN EN ISO 10519:2015-12 3 National Annex NA (informative) Bibliography DIN EN ISO 542, Oilseeds Sampling DIN EN ISO 648, Laboratory glassware Single-volume pipettes DIN EN ISO 664, Oilseeds Reduction of laboratory sample to test sample DIN EN ISO 665, Oilseeds Determination
11、 of moisture and volatile matter content DIN ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions DIN EN ISO 10519:2015-12 4 This page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 10519 Septembe
12、r 2015 ICS 67.200.20 Supersedes EN ISO 10519:2000English Version Rapeseed - Determination of chlorophyll content - Spectrometric method (ISO 10519:2015) Graines de colza - Dtermination de la teneur en chlorophylle - Mthode spectromtrique Rapssamen - Bestimmung des Chlorophyllgehaltes - Spektrometris
13、ches Verfahren (ISO 10519:2015) This European Standard was approved by CEN on 5 September 2015. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-d
14、ate lists and bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member. This European Standard exists in three official versions (English, French, German). A version in any other language made by translatio
15、n under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Fo
16、rmer Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey andUnited Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT
17、 EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2015 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 10519:2015 E(ISO 10519:2015) Foreword .1 Scope .2 No
18、rmative references 3 Terms and definitions .4 Principle 5 Reagent .5.1 Extraction solvent .6 Apparatus .7 Sampling 8 Preparation of test sample .9 Procedure.9.1 Test portions .9.2 Extraction 9.3 Determination .10 Expression of results 11 Precision .11.1 Repeatability .11.2 Reproducibility .12 Test r
19、eport .Annex A (informative) Results of interlaboratory test Bibliography .Contents PageEuropean foreword. . . 3 EN ISO 10519:2015 (E) 24555555566777778888910DIN EN ISO 10519:2015-12 European foreword This document (EN ISO 10519:2015) has been prepared by Technical Committee ISO/TC 34 “Food products
20、” in collaboration with Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products - Methods of sampling and analysis” the secretariat of which is held by AFNOR. This European Standard shall be given the status of a national standard, either by publication of
21、an identical text or by endorsement, at the latest by March 2016, and conflicting national standards shall be withdrawn at the latest by March 2016. 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 be h
22、eld responsible for identifying any or all such patent rights. This document supersedes EN ISO 10519:2000. 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, Cro
23、atia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and
24、the United Kingdom. Endorsement notice The text of ISO 10519:2015 has been approved by CEN as EN ISO 10519:2015 without any modification. EN ISO 10519:2015 (E) 3DIN EN ISO 10519:2015-12 ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards b
25、odies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizati
26、ons, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for its furthe
27、r maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives
28、). Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be in the Intr
29、oduction and/or on the ISO list of patent declarations received (see www.iso.org/patents). Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expressions related to
30、conformity assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for this document is ISO/TC 34, Food products, Subcommittee SC 2, Oleaginous seeds an
31、d fruits and oilseed meals.This third edition cancels and replaces the second edition (ISO 10519:1997), of which it constitutes a minor revision.Annex A of this International Standard is for information only.EN ISO 10519:2015 (E) 4DIN EN ISO 10519:2015-12 1 ScopeThis International Standard specifies
32、 a spectrometric method for the determination of the chlorophyll content of rapeseed. It is not applicable to the determination of chlorophyll in oils.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its applica
33、tion. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 648, Laboratory glassware Single-volume pipettesISO 664, Oilseeds Reduction of laboratory sample to test sampleISO 665, Oilseeds De
34、termination of moisture and volatile matter content3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1chlorophyll contentmass fraction of substances in the sample contributing to the absorption band at a wavelength near 665 nm, as determined under
35、the operating conditions specified in this International Standard and measured as chlorophyll ANote 1 to entry: The chlorophyll content is expressed in milligrams per kilogram.4 PrincipleExtraction of a test portion in a suitable apparatus with a specified extraction solvent. Spectrometric determina
36、tion of the chlorophyll content of the extracted solution.5 ReagentUse only reagents of recognized analytical grade unless otherwise stated.5.1 Extraction solventTransfer to a 500 ml beaker 100 ml of anhydrous ethanol. Add to the contents of the beaker 300 ml of anhydrous iso-octane (2,2,5-trimethyl
37、pentane) or anhydrous technical n-heptane or anhydrous petroleum ether (essentially composed of C7hydrocarbons, with a boiling range between 90 C and 100 C).6 ApparatusUsual laboratory apparatus and, in particular, are the following.EN ISO 10519:2015 (E) 5DIN EN ISO 10519:2015-12 6.1 Analytical bala
38、nce, capable of weighing to the nearest 0,001 g.6.2 Mechanical grinder, blade type, or coffee mill or equivalent.6.3 Mechanical microgrinder (see Figure 1), comprising stainless-steel tubes of approximately 50 ml volume which can be securely stoppered, stainless-steel ball-bearings ( 16 mm), and an
39、apparatus to shake the securely stoppered tubes horizontally at a frequency of 240 min1, with a horizontal displacement of 3,5 cm, or a commercial ball mill.Dimensions in millimetresKey1 neoprene or fluorosilicone stopper (No. 3)2 steel casing3 stainless-steel bearings, 16 mmFigure 1 Mechanical micr
40、ogrinder6.4 Filter paper, medium speed, V-folded.6.5 Spectrometer (preferably with wavelength scanning), suitable for carrying out absorbance measurements at wavelengths between 600 nm and 700 nm, with a spectral bandwidth of 2 nm.6.6 Optical cells, having a path length of at least 1 cm.6.7 Pipettes
41、, of 30 ml capacity, complying with the requirements of ISO 648, class A, or a repetitive dispenser capable of dispensing 30 ml with an error of less than 1 %.6.8 Culture tubes, of 20 ml capacity, provided with stoppers.7 SamplingSampling is not part of the method specified in this International Sta
42、ndard. A recommended sampling method is given in ISO 542.It is important that the laboratory receive a sample which is truly representative and has not been damaged or changed during transport and storage.8 Preparation of test samplePrepare a test sample in accordance with ISO 664 from the laborator
43、y sample as received, after separation of impurities.EN ISO 10519:2015 (E) 6DIN EN ISO 10519:2015-12 Dry seeds with a moisture content of greater than 10 % (m/m) for 12 h at 45 C to reduce the moisture level to 10 % (m/m) or less in order to reduce the risk of destroying chlorophyll pigments.Transfe
44、r 50 g of the test sample to the mechanical grinder (6.2) and grind to produce a uniformly ground seed. If a small grinder such as a coffee mill is used, grind several portions of 10 g and then combine them and mix thoroughly the ground portions.9 Procedure9.1 Test portionsWeigh, to the nearest 0,00
45、1 g, 2 g of the test sample (Clause 8) into a stainless-steel tube or the extraction vessel of a commercial ball mill (6.3).9.2 Extraction9.2.1 Add, using a pipette (6.7), 30 ml of the extraction solvent (5.1) to the tube or vessel. If using a tube, add three stainless-steel balls to the tube and sh
46、ake for 1 h. For commercial ball mills, add at least four medium-sized steel balls to the vessel and extract for 20 min.9.2.2 Allow the extract to settle for 10 min and then decant a sufficient volume of the extract through the filter paper (6.4) into a culture tube (6.8) to fill the optical cell (6
47、.6). Stopper the tube as soon as possible to minimize evaporation.NOTE The presence of more than one phase in the extraction solvent indicates the presence of excessive moisture, either in the sample which should contain less than 10 % (m/m) moisture or in the solvents (which should be anhydrous).9.
48、3 DeterminationTransfer the filtered extract to a cell (6.6) and determine by means of the spectrometer (6.5) the absorbance at wavelengths of 665 nm, 705 nm and 625 nm. (The readings at 705 nm and 625 nm are used to calculate a baseline correction.)10 Expression of resultsThe chlorophyll content, w
49、, in milligrams per kilogram of the product as received, is given by Formula (1):wkA Vml=corr(1)whereAcorr(the corrected absorbance) is equal to A665- (A705+ A625)/2;A665is the absorbance at 665 nm;A705is the absorbance at 705 nm;A625is the absorbance at 625 nm;k is a constant which is equal to 13;l is the path length, in centimetres, of the optical cell;m is the mass, in grams, of the test portion;EN ISO 10519:2015 (E) 7DIN EN ISO 10519:2015-12 V is the volume, in millilitres, of solvent added to the tube (9.2.1).If it is desire
