1、BSI Standards PublicationBS EN ISO 10504:2015Starch derivatives Determination of thecomposition of glucosesyrups, fructose syrupsand hydrogenated glucosesyrups Method usinghigh-performance liquidchromatography (ISO10504:2013)BS EN ISO 10504:2015 BRITISH STANDARDNational forewordThis British Standard
2、 is the UK implementation of EN ISO10504:2015. It is identical to ISO 10504:2013. It supersedes BS EN ISO10504:2000 which is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee AW/100, Agriculture and food.A list of organizations represented on this committee can be
3、obtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2015. Published by BSI StandardsLimited 2015ISBN 978 0 580 88370 5ICS 67.180.20Compliance w
4、ith a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 31 July 2015.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 1050
5、4 July 2015 ICS 67.180.20 Supersedes EN ISO 10504:2000English Version Starch derivatives - Determination of the composition of glucose syrups, fructose syrups and hydrogenated glucose syrups - Method using high-performance liquid chromatography (ISO 10504:2013) Produits drivs de lamidon - Dterminati
6、on de la composition des sirops de glucose, des sirops de fructose et des sirops de glucose hydrogns - Mthode par chromatographie en phase liquide haute performance (ISO 10504:2013) Strkederivate - Bestimmung der Zusammensetzung von Glucosesirup, Fructosesirup und hydriertem Glucosesirup -Hochleistu
7、ngs-flssigchromatographisches Verfahren (ISO 10504:2013) This European Standard was approved by CEN on 19 June 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 an
8、y alteration. Up-to-date 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 langua
9、ge made by translation 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,
10、 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 United Kingdom. EUROPEAN COMMITTEE FOR
11、 STANDARDIZATION COMIT 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 10504:2015 EBS EN ISO 10504:20
12、15EN ISO 10504:2015 (E) 3 European foreword The text of ISO 10504:2013 has been prepared by Technical Committee ISO/TC 93 “Starch (including derivatives and by-products)” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 10504:2015 by CCMC. This European S
13、tandard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by January 2016, and conflicting national standards shall be withdrawn at the latest by January 2016. Attention is drawn to the possibility that some of the elements
14、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. This document supersedes EN ISO 10504:2000. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following c
15、ountries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
16、 Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 10504:2013 has been approved by CEN as EN ISO 10504:2015 without any modification. BS EN ISO 10504:2015ISO 10504:2013(E) ISO 2013 All rights reserved iiiConte
17、nts PageForeword iv1 Scope . 12 Normative references 13 Principle 14 Reagents 15 Apparatus . 26 Procedure. 36.1 Choice of column . 36.2 System start-up . 36.3 Calibration of column 36.4 Sample preparation 46.5 Sample analysis 47 Calculation 48 Precision . 58.1 Repeatability . 58.2 Reproducibility .
18、5Annex A (informative) Examples of standard solutions . 7BS EN ISO 10504:2015ISO 10504:2013(E)ForewordISO (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
19、 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 organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO
20、 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 further maintenance are described in the ISO/IEC Directives, Part 1. In particular the different a
21、pproval 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. www.iso.org/directivesAttention is drawn to the possibility that some of the elements of this document may be the subj
22、ect 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 Introduction and/or on the ISO list of patent declarations received. www.iso.org/patentsAny trade name
23、used in this document is information given for the convenience of users and does not constitute an endorsement.The committee responsible for this document is ISO/TC 93, Starch (including derivatives and by-products).This second edition cancels and replaces ISO 10504:1998, of which it constitutes a m
24、inor revision.iv ISO 2013 All rights reservedBS EN ISO 10504:2015INTERNATIONAL STANDARD ISO 10504:2013(E)Starch derivatives Determination of the composition of glucose syrups, fructose syrups and hydrogenated glucose syrups Method using high-performance liquid chromatography1 ScopeThis International
25、 Standard describes a high-performance liquid chromatographic (HPLC) method for measuring the composition of dextrose solutions, glucose syrups, fructose-containing syrups, hydrogenated glucose syrups, sorbitol, mannitol and maltitol. The constituents are mainly glucose, maltose, maltotriose, fructo
26、se, sorbitol, mannitol, maltitol and malto-oligosaccharides.The use of a column packed with cation-exchange resin is essential.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated referenc
27、es, only the edition cited applies. 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 methodsISO 5381:1983, Starch hydrolysis products Determination of water content Modif
28、ied Karl Fischer method3 PrincipleSaccharide components are separated using high-performance liquid chromatography. Separation is achieved using a cation-exchange column with water as the eluent. The eluted components are detected by means of a differential refractometer, and quantified using an ele
29、ctronic integrator.4 ReagentsAll reagents used shall be of recognized analytical reagent grade.4.1 Special distilled water. The water used may be double-distilled of quality grade 1 in accordance with ISO 3696. The most suitable is demineralized water, which prevents contamination of the ion-exchang
30、e resin.The water should be filtered by passage through a 0,22 m filter. Also, it should be degassed by treatment under vacuum, or by use of an in-line degassing unit. The water should be maintained under an inert atmosphere, and preferably at 70 C to inhibit microbial growth.NOTE Some commercial wa
31、ter-purification devices produce water which is both filtered and degassed.4.2 Primary standard solutions.Prepare solutions (see Annex A) containing 10 % (or less) dry matter, according to the sensitivity of the refractometer, with compositions as close as possible to that of the samples to be analy
32、sed.NOTE Suitable reference materials for the constituents listed in Clause 1 can be obtained from established chemical companies. ISO 2013 All rights reserved 1BS EN ISO 10504:2015ISO 10504:2013(E)4.3 Ion-exchange resins, for off-line demineralization of samples.Salts present in the sample will co-
33、elute from the column, and will be detected by the refractometer, causing errors in the determination. These salts shall first be removed by ion-exchange resins. The most convenient way is to have an in-line guard column cartridge system (5.5), but this may also be carried out off-line using the fol
34、lowing resins1):a) Cation type:1) strong cation exchanger, 4 % cross-linked polystyrene divinylbenzene, in the H+form;2) 200 mesh to 400 mesh in the dry form;b) Anion type:1) weak anion exchanger, 4 % cross-linked polystyrene divinylbenzene support containing tertiary amine groups, in the free base
35、form;2) 200 mesh to 400 mesh in the dry form.5 Apparatus5.1 Liquid chromatograph; equipped with the following.5.1.1 Pump, pulseless, that delivers a constant flow, at the rate required.5.1.2 Differential refractometer, thermostatically controlled.5.1.3 Thermostatically controlled column oven, capabl
36、e of maintaining the column at temperatures up to 95 C, to within 0,5 C.5.2 Sample injector, comprising a loop injector (manual or part of autosampler) with a capacity of 20 l or less.5.3 Integrator, comprising an electronic integrator with calculating and recording capabilities, compatible with the
37、 voltage output of the detector.5.4 Separation column, comprising a pre-packed cation-exchange column in the form best suited for the analysis. The recommended resin is 6 % to 8 % cross-linked sulfonated polystyrene divinylbenzene with a bead diameter of 9 m to 25 m.NOTE Acceptable columns are avail
38、able from several major column suppliers.5.5 Guard columns, custom-prepared dual-cartridge system, inserted unheated in-line, to demineralize the sample.2)5.6 Sample filtration system, comprising a syringe to which suitable membrane disc filters can be attached. These should be of 0,45 m pore size.C
39、ommercially available syrups are usually highly refined, and a 0,45 m filter is suitable. However, if blockage of the chromatograph is too frequent, a 0,22 m filter should be used.1) While resins meeting these specifications are available from more than one supplier, their performance is variable. E
40、xperience in several laboratories has shown that the resins AG50W-X4 and AG3-X4 perform satisfactorily. (AG50W-X4 and AG3-X4 are trade names of products supplied by Bio-Rad. This information is given for the convenience of the users of this International Standard and does not constitute an endorseme
41、nt by ISO of these products. Equivalent products may be used if they can be shown to lead to the same results.)2) There are a few systems available but with varying efficiency. The Bio-Rad guard cartridges 1250118 have been shown in several laboratories to be the most effective in all respects. (Thi
42、s information is given for the convenience of the users of this International Standard and does not constitute an endorsement by ISO of these products. Equivalent products may be used if they can be shown to lead to the same results.)2 ISO 2013 All rights reservedBS EN ISO 10504:2015ISO 10504:2013(E
43、)6 Procedure6.1 Choice of columnFor general applications, a cation-exchange resin in the calcium form should be used, in particular for fructose syrups and hydrogenated glucose syrups. However, the separation of maltose at a high content from maltotriose is difficult when the maltotriose content is
44、about 6 % or more. In such instances better resolution is achieved with a cation-exchange resin in either the potassium or sodium form.6.2 System start-upInstall the column in the oven, and connect the guard columns (5.5) (if used) to the inlet. It is not necessary to heat the guard columns. Connect
45、 the injector to the inlet of the column (or guard columns, if used), and connect the outlet of the column to the detector inlet. Arrange that the detector effluent goes to waste.Start the pump at a rate of 0,1 ml/min, and pass the solvent through the column. Set the correct temperature for the colu
46、mn according to the suppliers recommendations. Enter the control parameters into the integrator. When the column temperature is stable, increase the solvent flow rate to 0,5 ml/min and purge the reference cell. Refer to the refractometer instruction manual to set the detector for correct measurement
47、 of the signal from the sample cell. Set the required attenuation.6.3 Calibration of column6.3.1 In accordance with the method specified in ISO 5381, determine the water content of every separate substance to be used for preparing the mixed primary standard solutions (see Annex A).For higher polyols
48、 (tri-itol and above), no commercial standards are available.6.3.2 Prepare a standard solution of each separate substance (see 4.2) and, using the same conditions as those to be used for the analysis, inject an aliquot portion several times into the column. At least three results, based on integrato
49、r response, should show a variation of 0,1 % or less for the major constituent. Calculate an average result for all components.NOTE For the single primary substances, an assumption is made that each sugar has the same relative response, and that the normalized area percentage figures reflect the true analysis. To obtain the required level of higher molecular weight species, a dextrin, or a fraction especially prepared from a starch hydrolysate, can be used.6.3.3 Prepare mixtures of the single substances to give compositions a
