DIN EN ISO 13906-2008 Animal feeding stuffs - Determination of acid detergent fibre (ADF) and acid detergent lignin (ADL) contents (ISO 13906 2008) English version of DIN EN ISO 13.pdf

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1、November 2008DEUTSCHE NORM English price group 12No part of this standard 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 65.120!$Rde“1476566ww

2、w.din.deDDIN EN ISO 13906Animal feeding stuffs Determination of acid detergent fibre (ADF) and acid detergent lignin(ADL) contents (ISO 13906:2008)English version of DIN EN ISO 13906:2008-11Futtermittel Bestimmung des Gehalts an Sure-Detergens-Faser (ADF) und Sure-Detergens-Lignin(ADL) (ISO 13906:20

3、08)Englische Fassung DIN EN ISO 13906:2008-11www.beuth.deDocument comprises 22 pagesDIN EN ISO 13906:2008-11 2 National foreword This standard has been prepared by Technical Committee ISO/TC 34 “Food products” (Secretariat: AFNOR, France), Subcommittee SC 10 “Animal feeding stuffs” (Secretariat: NEN

4、, Netherlands) in collaboration with Technical Committee CEN/TC 327 “Animal feeding stuffs Methods of sampling and analysis” (Secretariat: NEN, Netherlands). Based on the results of parallel voting, ISO 13906:2008 has been adopted as a European Standard. The responsible German body involved in its p

5、reparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Technical Committee NA 057-03-03 AA Futtermittel. The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 572

6、5-1 DIN ISO 5725-1 ISO 5725-2 DIN ISO 5725-2 ISO 6497 DIN EN ISO 6497 National Annex NA (informative) Bibliography DIN EN ISO 6497, Animal feeding stuffs Sampling DIN ISO 5725-1, Accuracy (trueness and precision) of measurement methods and results Part 1: General principles and definitions DIN ISO 5

7、725-2, 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 EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 13906July 2008ICS 65.120English VersionAnimal feeding stu

8、ffs - Determination of acid detergent fibreAliments des animaux - Dtermination des teneurs enfibres au dtergent acide (ADF) et en lignine sulfurique(ADL) (ISO 13906:2008)Futtermittel - Bestimmung des Gehalts an Sure-Detergens-Faser (ADF) und Sure-Detergens-Lignin (ADL)(ISO 13906:2008)This European S

9、tandard was approved by CEN on 11 July 2008.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning s

10、uch nationalstandards 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, German). A version in any other language made by translationunder the responsibility of a CEN member into its own language

11、 and notified to the CEN Management Centre has the same status as theofficial versions.CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland,France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembou

12、rg, Malta, Netherlands, Norway, Poland, Portugal,Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOMITEE FR NORMUNGManagement Centre: rue de Stassart, 36 B-1050 Brussels 2008 CEN All rights of

13、exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN ISO 13906:2008: E(ADF) and acid detergent lignin (ADL) contents(ISO 13906:2008)Contents Page 1 2 3 4 5 6 7 8 9 10 11 12 DIN EN ISO 13906:2008-11 EN ISO 13906:2008 (E)2 Foreword. 3 Scope 4 Normative refer

14、ences 4 Terms and definitions .4 Principle5 Reagents.5 Apparatus .6 Sampling.7 Preparation of test sample7 Procedure .7 Calculation and expression of results.9 Precision.10 Test report 12 Annex A (informative) Results of interlaboratory test.13 Annex B (informative) Results of interlaboratory test.1

15、7 Bibliography 20 Foreword 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 January 2009, and conflicting national standards shall be withdrawn at the latest by January 2009. Attention is drawn to

16、the possibility that some of the elements of this document may be the subject of patent rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following cou

17、ntries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Sp

18、ain, Sweden, Switzerland and the United Kingdom. Endorsement notice The text of ISO 13906:2008 has been approved by CEN as a EN ISO 13906:2008 without any modification. This document (EN ISO 13906:2008) has been prepared by Technical Committee ISO/TC 34 “Food products” in collaboration with Technica

19、l Committee CEN/TC 327 “Animal feeding stuffs - Methods of sampling and DIN EN ISO 13906:2008-11 EN ISO 13906:2008 (E)3 analysis” the secretariat of which is held by NEN. WARNING The use of this International Standard may involve hazardous materials, operations and equipment. This International Stan

20、dard does not purport to address any safety risks associated with its use. It is the responsibility of the user of this International Standard to establish appropriate safety and health practices and determine the applicability of local regulatory limitations prior to use. 1 Scope This International

21、 Standard specifies a method for the determination of acid detergent fibre (ADF) insoluble residue and acid detergent lignin (ADL) in all types of animal feeding stuffs. The limit of determination is 1 % mass fraction for ADF and 1,5 % mass fraction for ADL. A gravimetric routine and reference metho

22、d is used. 2 Normative references The following referenced documents 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. ISO 6498, Anima

23、l feeding stuffs Preparation of test samples 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 acid detergent fibre content ADF content mass fraction of fibrous residue obtained after treatment with cationic detergent in 0,5 mol/l sulfuric acid

24、, primarily consisting of cellulose, lignin and insoluble protein complexes NOTE The ADF mass fraction is expressed as a percentage. 3.2 acid detergent lignin content ADL content mass fraction of residue remaining after cellulose and other organic matter is solubilized by 72 % mass fraction (12,00 m

25、ol/l) sulfuric acid NOTE The ADL mass fraction is expressed as a percentage. DIN EN ISO 13906:2008-11 EN ISO 13906:2008 (E)4 4 Principle ADF is determined in the first stage of the method. Cationic detergent solution is used to remove acid-labile carbohydrates, protein that is not complexed into Mai

26、llard products (heat damaged), and fats. The remaining fibrous residue is primarily cellulose and lignin (plant products) or insoluble protein complexes (animal products and heat-damaged feeds). The residue is weighed for the determination of ADF. In the second stage, the remaining residue is solubi

27、lized by 72 % mass fraction (12,00 mol/l) sulfuric acid, leaving the lignin (ADL) which is determined gravimetrically. 5 Reagents Use only reagents of recognized analytical grade, unless otherwise specified, and only distilled or deionized water or water of at least equivalent purity. 5.1 Acid deter

28、gent solution. Add 20 g cetyl(trimethyl)ammonium bromide (technical grade) to 1 l of 0,5 mol/l sulfuric acid, previously standardized. Agitate to aid dissolution. 5.2 Sulfuric acid, 72 % mass fraction (12,00 mol/l). Standardize sulfuric acid (H2SO4) to a relative density of 1,634 at 20 C or 12,00 mo

29、l/l as follows. Calculate the mass, in grams, of acid, m(H2SO4), and the mass, in grams, of water, m(H2O), needed to prepare 1 000 ml of solution using Equations (1) and (2): 2424100 98,08 12(H SO )(H SO )mw= (1) where w(H2SO4) is the assay mass fraction of sulfuric acid, expressed as a percentage.

30、( )224(H O) 1000 1,634 (H SO )mm= (2) where 1,634 is the relative density of 72 % mass fraction sulfuric acid. Weigh water into a 1 000 ml volumetric flask and add the calculated amount of sulfuric acid slowly with occasional swirling. Cool the flask in a water bath while adding the required mass of

31、 acid. Cool to 20 C and verify the volume. The meniscus should be within 0,5 cm of the calibration mark at 20 C. If volume is too large, remove 5 ml water and add 4,55 ml sulfuric acid. If volume is too small, remove 1,5 ml and add 2,5 ml water. Repeat if necessary. 5.3 Filtration aid, diatomaceous

32、earth1). 5.4 Acetone, technical grade. 5.5 n-Octanol, antifoaming agent. 1) Celite, acid washed, and Celite 545 AW are examples of suitable products available commercially. This information is given for the convenience of users of this International Standard, and does not constitute an endorsement o

33、f these products by ISO. Alternative products may be used if they can be demonstrated to give comparable results. DIN EN ISO 13906:2008-11 EN ISO 13906:2008 (E)5 6 Apparatus Usual laboratory apparatus and, in particular, the following. 6.1 Analytical balance. 6.1.1 Analytical balance, capable of wei

34、ghing to the nearest 1 mg, with readability to 0,1 mg. 6.1.2 Analytical balance, with a measuring range up to 2 500 g, with readability to 1 g, for weighing sulfuric acid (5.2). 6.2 Mill, a cyclone mill or cutter mill or rotary mill or equivalent, giving a mean particle size of 0,22 mm to 0,26 mm. 6

35、.3 Drying oven. 6.3.1 Air-ventilated oven, capable of operating at 103 C 2 C or 130 C 2 C. 6.3.2 Air-ventilated oven, capable of being maintained at 60 C 2 C. To speed up the drying of moist samples without creating artefact fibres, a vacuum oven, maintained at 60 C 2 C may also be used. 6.4 Refluxi

36、ng apparatus, with individual heating units and cold water condensers. Any conventional apparatus suitable for crude fibre or amylase-treated neutral detergent fibre (aNDF) determinations is acceptable. Calibrate heating unit settings so that 50 ml of water boils in 4 min to 5 min when using cold wa

37、ter condensers. A Fibertec2)-type apparatus can be used and should boil 50 ml of water within 10 min. NOTE This setting can be expected to result in significant particle movement during refluxing. 6.5 Fritted-disk crucibles. Coarse porosity (pore size 40 m to 60 m) crucibles, 40 ml to 50 ml capacity

38、, or Fibertec P22)(pore size 40 m to 100 m, 26 ml to 28 ml capacity). Clean new crucibles and ash at 525 C 15 C for 1 h. Clean crucibles after each use by ashing at 525 C 15 C for 3 h, removing ash by inverting in a detergent solution and sonicating for 7 min to 10 min. Rinse crucibles in hot water,

39、 and soak in water at room temperature for at least 30 min. Occasionally test filtration rate as follows. Fill each crucible with 50 ml of distilled water (25 ml for Fibertec P22)crucibles) and record the time required to drain completely without vacuum should be 180 s 60 s for Gooch2)or 75 s 30 s f

40、or P2. If drain time is 240 s (or 105 s for P2), clean crucible with acid or alkaline cleaning solution (Reference 4). If cleaning does not improve filtration rate, discard crucible. 6.6 Vacuum filter manifold. Suitable apparatus e.g. Fibertec2)-type cold extraction unit that allows adequate soaking

41、 of fibrous residues. 6.7 Incineration furnace, 525 C 15 C. 6.8 Reflux beakers. As an alternative to a reflux apparatus (6.4), 600 ml Berzelius beakers with condensers, e.g. made from 500 ml round-bottom flasks, may be used. 2) Example of a suitable product available commercially. This information i

42、s given for the convenience of users of this International Standard, and does not constitute an endorsement of this product by ISO. DIN EN ISO 13906:2008-11 EN ISO 13906:2008 (E)6 7 Sampling A representative sample should have been sent to the laboratory. It should not have been damaged or changed d

43、uring transport or storage. Sampling is not a part of the method specified in this International Standard. A recommended sampling method is given in ISO 6497. 8 Preparation of test sample Prepare the test sample in accordance with ISO 6498. Reduce laboratory sample to approximately 100 g (dry weight

44、 equivalent) and place half in a moisture-tight, sealed container for total moisture determination. Dry remaining wet ( 15 % mass fraction moisture) materials to 10 % mass fraction fat, and is recommended for test samples containing 5 % mass fraction fat. Weigh test samples into previously tared, ov

45、en-dried crucibles, place crucible on filtering manifold, extract four times with 30 ml to 40 ml of acetone, allowing the material to soak in acetone for 3 min to 5 min each time, vacuum to remove all traces of acetone, air-dry for 10 min to 15 min, and transfer residue to a reflux beaker for fibre

46、analysis. Use the same crucible to collect the fibre residue for each test portion after acid detergent extraction. For the Fibertec2)-type apparatus, place the crucible in a cold extraction unit and fill the crucible with 25 ml acetone (5.4). Leave for 3 min to 5 min and filter by applying vacuum.

47、Repeat three times. NOTE To simplify filtration, 1,00 g of filtration aid (5.3) can be added to the crucible before the sample. 9 Procedure 9.1 ADF 9.1.1 General Dry empty crucibles in an oven (6.3.1) at 103 C 2 C for 4 h 1 h if moved from the incineration furnace (6.7) and record tare mass, m1. 9.1

48、.2 Conventional apparatus Weigh, into a Berzelius beaker (6.8), a test portion of dried or as-received material of mass 1 000 mg 2 mg, and record the mass as m2. Materials with 15 % mass fraction moisture should have the mass adjusted to provide an equivalent mass of dry matter. If conversion of res

49、ults to the dry matter basis is required, weigh a test portion for moisture determination of the test sample at the same time. Immediately before refluxing, add 100 ml of acid detergent solution (5.1) at room temperature. Heat to boiling over 5 min to 10 min and, if necessary, reduce heat slightly and add 2 drops to 4 drops of n-octanol (5.5) to avoid foaming, but provide moderate particle agitation. After 5 min to 10 min of refluxing, rinse down the sides of the beaker using a fine stream of acid deterge

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