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1、| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS EN ISO 10519:2001 BS 4
2、289-10:1998 Incorporating Amendment No. 1 to BS 4289-10:1998 (renumbers the BS as BS EN ISO 10519:2001) and Corrigendum No. 1 to BS EN ISO 10519:2001 The European Standard EN ISO 10519:2000 has the status of a British Standard ICS 67.200.20 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY CO
3、PYRIGHT LAW Rapeseed Determination of chlorophyll content Spectrometric method Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy, (c) BSI This British Standard, having been prepared under the direction of the Agriculture and Food Standards Policy Commi
4、ttee, was published under the authority of the Standards Committee and comes into effect on 15 October 1992 BSI 03-2001 The following BSI references relate to the work on this standard: Committee reference AFC/2 Draft for comment 91/52788 DC ISBN 0 580 21273 4 BS EN ISO 10519:2001 Amendments issued
5、since publication Amd. No.DateComments 11091February 2001 Implementation of the European Standard 13102 Corrigendum March 2001ISO text corrected National foreword This British Standard is the official English language version of EN ISO 10519:2000. It is identical with ISO 10519:1997. The UK particip
6、ation in its preparation was entrusted to Technical Committee AW/2, Oilseeds and residues, which has the responsibility to: aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK
7、interests informed; monitor related international and European developments and promulgate them in the UK. A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications
8、referred to in this document may be found in the BSI Standards Catalogue under the section entitled International Standards Correspondence Index, or by using the Find facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a
9、contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, the EN ISO title page, the EN ISO foreword
10、page, the ISO title page, page ii, pages 1 to 7, an annex ZA page, an inside back cover and a back cover. The BSI copyright notice displayed in this document indicates when the document was last issued. Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy
11、, (c) BSI EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 10519 August 2000 ICS 67.200.20 English version Rapeseed - Determination of chlorophyll content - Spectrometric method (ISO 10519:1997) Graines de colza - Dtermination de la teneur en chlorophylle - Mthode spectromtrique (ISO 10519:1
12、997) Rapssamen - Bestimmung des Chlorophyllgehaltes - Spektrometrisches Verfahren (ISO 10519:1997) This European Standard was approved by CEN on 5 August 2000. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard t
13、he 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 Central Secretariat or to any CEN member. This European Standard exists in three official versions (English, French, Germa
14、n). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Fin
15、land, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Central Secretariat: rue de Stassart, 36 B-1050 Bruss
16、els 2000 CENAll rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 10519:2000 E Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy, (c) BSI Foreword The text of the International Standard fro
17、m Technical Committee ISO/TC 34 “Agricultural food products“ of the International Organization for Standardization (ISO) has been taken over as an European Standard by Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products - Methods of sampling and analysi
18、s“, the secretariat of which is held by AFNOR. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by February 2001, and conflicting national standards shall be withdrawn at the latest by February 2001.
19、According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Por
20、tugal, Spain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of the International Standard ISO 10519:1997 has been approved by CEN as a European Standard without any modification. NOTE: Normative references to International Standards are listed in annex ZA (normative). EN IS
21、O 10519:2000 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy, (c) BSI A Reference number ISO 10519:1997(E) INTERNATIONAL STANDARD ISO 10519 Second edition 1997-11-01 Rapeseed Determination of chlorophyll content Spectrometric method Graines de colza
22、Dtermination de la teneur en chlorophylle Mthode spectromtrique EN ISO 10519:2000 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy, (c) BSI )E(7991:91501 OSI ii Foreword ISO (the International Organization for Standardization) is a worldwide federatio
23、n of national standards bodies (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.
24、 International organizations, 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. Draft International Standards adopted by the technic
25、al committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. International Standard ISO 10519 was prepared by Technical Committee ISO/TC 34, Agricultural food products, Subcommittee SC 2
26、, Oleaginous seeds and fruits. This second edition cancels and replaces the first edition (ISO 10519:1992), which has been technically revised. Annexes A and B of this International Standard are for information only. EN ISO 10519:2000 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 G
27、MT+00:00 2006, Uncontrolled Copy, (c) BSI DRADNATS LANOITANRETNI OSIOSI )E(7991:91501 1 Rapeseed Determination of chlorophyll content Spectrometric method 1Scope This International Standard specifies a spectrometric method for the determination of the chlorophyll content of rapeseed. It is not appli
28、cable to the determination of chlorophyll in oils. 2Normative references The following standards contain provisions which, through reference in this test, constitute provisions of this International Standard. At the time of the publication, the editions indicated were valid. All standards are subjec
29、t to revision, and the parties to agreements based on this International Standard are encouraged to investigate the possibility of applying the most recent editions of the standards indicated below. Members of IEC and ISO maintain registers of currently valid International Standards. ISO 648:1977, L
30、aboratory glassware - One-mark pipettes. ISO 664:1990, Oilseeds - Reduction of laboratory sample to test sample. ISO 665:1977, Oilseeds - Determination of moisture and volatile matter content. 3Definition For the purposes of this International Standard, the following definition applies. 3.1 chloroph
31、yll content mass fraction of substances in the sample contributing to the absorption band at a wavelength near 665 nm, as determined under the operating conditions specified in this International Standard and measured as chlorophyll A NOTE The chlorophyll content is expressed in milligrams per kilog
32、ram. 4Principle Extraction of a test portion in a suitable apparatus with a specified extraction solvent. Spectrometric determination of the chlorophyll content of the extracted solution 5Reagent Use only reagents of recognized analytical grade unless otherwise stated. EN ISO 10519:2000 Licensed Cop
33、y: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy, (c) BSI )E(7991:91501 OSI OSI 2 5.1Extraction solvent Transfer 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-trimethylpentane) or anhydrous
34、 technical n-hep- tane or anhydrous petroleum ether (essentially composed of C7 hydrocarbons, with a boiling range between 90 C and 100 C). 6Apparatus Usual laboratory apparatus and, in particular, the following. 6.1Analytical balance, capable of weighing to the nearest 0,001 g. 6.2Mechanical grinde
35、r, blade type, or coffee mill or equivalent. 6.3Mechanical microgrinder (see figure 1), comprising stainless-steel tubes of approx- imately 50 ml volume which can be securely stoppered, stainless-steel ball-bearings ( 16 mm), and an apparatus to shake the securely stoppered tubes horizontally at a f
36、requency of 240 min1, with a horizontal displacement of 3,5 cm, or a Dangoumau ball mill1). Dimensions in millimetres Figure 1 Mechanical microgrinder 6.4Filter paper, medium speed, V-folded. 6.5Spectrometer (preferably with wavelength scanning), suitable for carrying out absorbance measurements at
37、wavelengths between 600 nm and 700 nm, with a spectral bandwidth of 2 nm. 6.6Optical cells, having a path length of at least 1 cm. 6.7Pipettes, 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 %
38、. 6.8Culture tubes, of 20 ml capacity, provided with stoppers. 1) Dangoumau ball mill is an example of a suitable apparatus available commercially. This information is given for the convenience of users of this International Standard and does not constitute an endorsement by ISO of this apparatus. E
39、N ISO 10519:2000 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy, (c) BSI OSI)E(7991:91501 OSI 3 7Sampling Sampling is not part of the method specified in this International Standard. A recommended sampling method is given in ISO 542. It is important
40、 that the laboratory receive a sample which is truly representative and has not been damaged or changed during transport and storage. 8Preparation of test sample Prepare a test sample in accordance with ISO 664 from the laboratory sample as received, after separation of impurities. Dry seeds with a
41、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. Transfer 50 g of the test sample to the mechanical grinder (6.2) and grind to produce a uniformly ground seed. If a small gr
42、inder such as a coffee mill is used, grind several portions of 10 g and then combine them and mix thoroughly the ground portions. 9Procedure 9.1Test portions Weigh, to the nearest 0,001 g, 2 g of the test sample (clause 8) into a stainless-steel tube or the extraction vessel of the Dangoumau ball mi
43、ll (6.3). 9.2Extraction 9.2.1 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 shake for 1 h. For Dangoumau ball mills, add at least four medium-sized steel balls to the vessel and extract for 20
44、 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 (6.6). Stopper the tube as soon as possible to minimize evaporation. NOTE The presence of more than one phase in the
45、 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). 9.3Determination Transfer the filtered extract to a cell (6.6) and determine by means of the spectrometer (6.5) th
46、e 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.) EN ISO 10519:2000 Licensed Copy: sheffieldun sheffieldun, na, Sun Nov 19 14:53:41 GMT+00:00 2006, Uncontrolled Copy, (c) BSI )E(7991:91501 OSI OSI 4 10Expression
47、 of results The chlorophyll content, w, in milligrams per kilogram of the product as received, is given by the formula w kAV ml = corr where Acorr (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
48、 625 nm; kis a constant which is equal to 13; lis the path length, in centimetres, of the optical cell; mis the mass, in grams, of the test portion; Vis the volume, in millilitres, of solvent added to the tube (9.2.1). If it is desired to express the chlorophyll content relative to the dry product,
49、take into account in the calculation the moisture content of the sample, determined in accordance with ISO 665. 11Precision Details of an interlaboratory test on the precision of the method are summarized in annex A. The values derived from this interlaboratory test may not be applicable to concentration ranges and matrices other than those given. 11.1 Repeatability The absolute difference between two independent single test results, obtained using the same method on identical test material in the same laboratory by the sam