1、BRITISH STANDARDBS EN ISO 15914:2005Incorporating Amendment No. 1 to BS ISO 15914:2004 (renumbers BS ISO as BS EN ISO 15914:2005)Animal feeding stuffs Enzymatic determination of total starch contentThe European Standard EN ISO 15914:2005 has the status of a British StandardICS 65.120g49g50g3g38g50g5
2、1g60g44g49g42g3g58g44g55g43g50g56g55g3g37g54g44g3g51g40g53g48g44g54g54g44g50g49g3g40g59g38g40g51g55g3g36g54g3g51g40g53g48g44g55g55g40g39g3g37g60g3g38g50g51g60g53g44g42g43g55g3g47g36g58BS EN ISO 15914:2005This British Standard was published under the authority of the Standards Policy and Strategy Com
3、mittee on 4 February 2004 BSI 17 February 2006ISBN 0 580 43337 4National forewordThis British Standard is the English language version of EN ISO 15914:2005. It is identical with ISO 15914:2004.The UK participation in its preparation was entrusted to Technical Committee AW/10, Animal feeding stuffs,
4、which has the responsibility to: A list of organizations represented on this committee can be obtained on request to its secretary.Cross-referencesThe British Standards which implement international or European publications referred to in this document may be found in the BSI Catalogue under the sec
5、tion entitled “International Standards Correspondence Index”, or by using the “Search” facility of the BSI Electronic Catalogue or of British Standards Online.This publication does not purport to include all the necessary provisions of a contract. Users are responsible for its correct application. C
6、ompliance with a British Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep UK interests informed; monitor related
7、 international and European developments and promulgate them in the UK.Summary of pagesThis document comprises a front cover, an inside front cover, the EN ISO title page, the EN ISO foreword page, the ISO title page, pages ii to iv, pages 1 to 12, an inside back cover and a back cover.The BSI copyr
8、ight notice displayed in this document indicates when the document was last issued.Amendments issued since publicationAmd. No. Date Comments15759 February 2006 Renumbers BS ISO 15914:2004 as BS EN ISO 15914:2005EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORMEN ISO 15914March 2005ICS 65.120English ve
9、rsionAnimal feeding stuffs Enzymatic determination of total starchcontent (ISO 15914:2004)Aliments des animaux Dtermination enzymatique de lateneur totale en amidon (ISO 15914:2004)Futtermittel Enzymatische Bestimmung von Strke (ISO15914:2004)This European Standard was approved by CEN on 7 March 200
10、5.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 such nationalstandards may be obtained on a
11、pplication to the Central Secretariat 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 and notified to the Central Secretariat has
12、 the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovak
13、ia,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 2005 CEN All rights of exploitation in any form and by any means reservedworldwide for C
14、EN national Members.Ref. No. EN ISO 15914:2005: E- Foreword The text of ISO 15914:2004 has been prepared by Technical Committee ISO/TC 34, Agricultural food products, of the International Organization for Standardization (ISO) and has been taken over as EN ISO 15914:2005 by Technical Committee CEN/T
15、C 327, Animal feeding stuffs Methods of sampling and analysis, the secretariat of which is held by NEN. 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 September 2005, and conflicting national sta
16、ndards shall be withdrawn at the latest by September 2005. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Ge
17、rmany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom. Endorsement notice The text of ISO 15914:2004 has been approved by CEN as EN ISO 15914:2005 without any mod
18、ifications. EN ISO 15914:2005Reference numberISO 15914:2004(E)OSI 4002INTERNATIONAL STANDARD ISO15914First edition2004-02-01Animal feeding stuffs Enzymatic determination of total starch content Aliments des animaux Dtermination enzymatique de la teneur totale en amidon EN ISO 15914:2005IS:41951 O400
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22、SO 4002 All irthgs erse.devr lnUeto sswrehise specified, on trap fo this lbupictaion maeb y cudorperro de tuilizi den yna form ro na ybm ynae,s lecetrinoc ro mecinahcal, inclidung tohpcoiypodna gn micrfoilm, wittuoh repmissii non writign from ietI rehSa Ot tsserdda eh ebolw or ISOs memreb i ydobn th
23、e cnuotrfo y ttseuqer ehe.r ISO cirypothg fofice saCe tsopale 65 eneG 1121-HC 02 avleT. 4 + 10 947 22 1 11 xaF0 947 22 14 + 9 74 E-mail coirypthgiso.o grWe bwww.is.o groPulbisdehi n Switlrez dnaii ISO 4002 Allr ithgsr esedevrI SO 4002 All irthgs ersedevr iiiContents Page Foreword iv 1 Scope 1 2 Norm
24、ative references . 1 3 Terms and definitions. 1 4 Principle . 2 5 Reagents 2 6 Apparatus. 4 7 Preparation of test sample. 5 8 Procedure. 5 9 Calculation and expression of results 7 10 Precision 8 11 Test report 9 Annex A (informative) Notes on procedure . 10 Annex B (informative) Results of interlab
25、oratory test 11 Bibliography . 12 EN ISO 15914:2005iv I SO 4002 All irthgs ersedevrForeword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out throu
26、gh 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 organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collabora
27、tes closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standar
28、ds. Draft International Standards adopted by the technical 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. Attention is drawn to the possibility that some of the elements of th
29、is document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. ISO 15914 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 10, Animal feeding stuffs. EN ISO 15914:2005INTENRATIONAL TSANDADR I SO 4002 All i
30、rthgs ersedevr 1Animal feeding stuffs Enzymatic determination of total starch content 1 Scope This international Standard specifies a method for the enzymatic determination of the total starch content of animal feeding stuffs and raw materials for animal feeding stuffs. The method is also applicable
31、 to the determination of the purity of starch. It is important that in the sample matrix no components are present which contribute to the measured extinction at 340 nm. The analytical range of the method is 40 g/kg to 1 000 g/kg starch. The standard procedure is applicable to the range 200 g/kg to
32、1000 g/kg. For the lower range, 40 g/kg to 200 g/kg, another dilution procedure for the standard glucose solution and samples can be used. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited ap
33、plies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 3696:1987, Water for analytical laboratory use Specification and test methods ISO 6498:1998, Animal feeding stuffs Preparation of test samples 3 Terms and definitions For the purpose
34、of this document, the following terms and definitions apply. 3.1 starch natural vegetable polymer consisting of long linear unbranched chains of 1,4-D-glucose units (amylose) and/or long -1,6-branched chains of -1,4-linked glucose units (amylopectin) 3.2 starch content mass fraction of starch and it
35、s high molecular mass breakdown products, insoluble in 40 % ethanol, and determined in accordance with the method of this International Standard NOTE The starch content is expressed in grams per kilogram. EN ISO 15914:20052 I SO 4002 All irthgs ersedevr4 Principle The milled test sample is extracted
36、 with 40 % ethanol to remove the soluble sugars. Sample disintegration and starch solubilization is achieved by dispersing the extracted test portion in aqueous DMSO (90 % volume fraction) at 100 C, followed by addition of concentrated hydrochloric acid at 60 C. The solubilized and dissolved starch
37、is quantitatively converted into glucose by the enzyme amyloglucosidase. The glucose quantification is carried out by the well-known hexokinase method (see 1, 2). 5 Reagents Use only reagents of recognized analytical grade. 5.1 Water, complying with at least grade 3 in accordance with ISO 3696:1987.
38、 5.2 Ethanol (C2H5OH), 40 % (volume fraction) Take 417 ml of ethanol (96 % volume fraction) and dilute with water to 1 000 ml 5.3 Hydrochloric acid, c(HCl) = 12 mol/l. 5.4 Aqueous sodium hydroxide, c(NaOH) = 4 mol/l. Dissolve in a beaker 40 g of NaOH in about 50 ml water. After cooling, transfer qua
39、ntitatively to a 250 ml volumetric flask and dilute to the mark with water. WARNING Heat develops. Wear safety glasses. 5.5 Acetic acid solution, c(CH3COOH) = 2 mol/l. Add to a 500 ml volumetric flask about 200 ml water, followed by 59 ml of glacial acetic acid. Dilute to the mark with water. 5.6 So
40、dium acetate solution, c(CH3COONa) = 2 mol/l. Dissolve, in a 500 ml volumetric flask, 82,0 g of sodium acetate in water and dilute to the mark with water. 5.7 Sodium acetate buffer, c(CH3COONa/H) = 2 mol/l, pH = 4,8. Mix 41 ml of acetic acid solution (5.5) with 59 ml of sodium acetate solution (5.6)
41、. Check the pH with a pH-meter and, if necessary, adjust to obtain the correct pH with the acetic acid or the sodium acetate solution. Prepare a fresh buffer solution daily. 5.8 Aqueous dimethylsulfoxide, (DMSO) = 90 % (volume fraction). Mix pure DMSO and water in a volume ratio of 9:1. 5.9 Clarifyi
42、ng solutions, according to Carrez, prepared as follows. 5.9.1 Potassium hexacyanoferrate(II) solution, cK4Fe(CN)6 = 0,25 mol/l. In a 1 litre volumetric flask, dissolve 106 g of potassium hexacyanoferrate(II) trihydrate K4Fe(CN)63H2O in water. Dilute to the mark with water. 5.9.2 Zinc acetate, soluti
43、on in 0,5 mol/l acetic acid, cZn(CH3CO2)2 = 1 mol/l. EN ISO 15914:2005I SO 4002 All irthgs ersedevr 3In a 1 litre volumetric flask, dissolve 219,5 g of zinc acetate dihydrate Zn(CH3CO2)22H2O and 30 g of glacial acetic acid in water. Dilute to the mark with water. 5.10 Iodine solution in potassium io
44、dide In a 1 litre volumetric flask, dissolve 12,7 g of iodine (I2) and 24,0 g of potassium iodide (KI) in water. Dilute to the mark with water. Dilute this solution 10-fold before use. 5.11 Standard glucose solution 5.11.1 Samples containing 200 g/kg to 1 000 g/kg starch Prepare three independent st
45、andard glucose solutions (c = 0,0194 mol/l). In each 100 ml volumetric flask dissolve 350 mg anhydrous glucose (C6H12O6) to the nearest mg in water. Dilute to the mark with water. 5.11.2 Samples containing 40 g/kg to 200 g/kg starch Prepare three independent standard glucose solutions (c = 0,0039 mo
46、l/l). In each 500 ml volumetric flask, dissolve 350 mg 1 mg of anhydrous glucose (C6H12O6) in water. Dilute to the mark with water. Prepare fresh standard glucose solutions daily. 5.12 Amyloglucosidase solution, 160 U/ml AMG. Dissolve, in a mixture of 9 ml of water and 1 ml of sodium acetate buffer
47、(5.7), 267 mg of amyloglucosidase (AMG) EC 3.2.1.3 (Aspergillus niger; Roche Diagnostics, No. 1 202 367, 6 U/mg).1)With respect to storage and the use of the enzymes, follow carefully the recommendations of the enzyme supplier. When another enzyme is used, the activity should be estimated as given i
48、n Note 1. The mass of the enzyme should be adapted to the activity found. NOTE 1 Different enzyme suppliers use different definitions for the units of activities of enzymes. In this International Standard the following definition for the unit of activity for AMG has been used: 1 unit of amyloglucosi
49、dase will release 1 mol of glucose from glycogen in 1 min at 25 C and pH = 4,75. NOTE 2 10 ml of enzyme solutions is enough for about 75 determinations. 5.13 D-Glucose UV test set, for quantifying glucose enzymaticly, with the hexokinase method (R-Biopharm, No. 7162512), according to the manufacturer, the unused kits may be stored for 1 year at 4 C), as given in 5.13.1 to 5.13.3. 5.13.1 Buffer/substrate solution (Bottle 1) Dissolve the content of Bottle 1 in 45 ml of freshly prepared distilled water. Sto
