DIN EN ISO 10504-2015 Starch derivatives - Determination of the composition of glucose syrups fructose syrups and hydrogenated glucose syrups - Method using high-performance liquid.pdf

1、November 2015 English price group 8No 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 67.180.20!%GNN“2364343www.din.de

2、DIN EN ISO 10504Starch derivatives Determination of the composition of glucose syrups, fructose syrups and hydrogenated glucose syrups Method using high-performance liquid chromatography (ISO 10504:2013);English version EN ISO 10504:2015,English translation of DIN EN ISO 10504:2015-11Strkederivate B

3、estimmung der Zusammensetzung von Glucosesirup, Fructosesirup und hydriertem Glucosesirup Verfahren mittels Hochleistungs-Flssigchromatographie (ISO 10504:2013);Englische Fassung EN ISO 10504:2015,Englische bersetzung von DIN EN ISO 10504:2015-11Produits drivs de lamidon Dtermination de la compositi

4、on 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);Version anglaise EN ISO 10504:2015,Traduction anglaise de DIN EN ISO 10504:2015-11SupersedesDIN EN ISO 10504:2000-08www.beuth.deDocument c

5、omprises 12 pagesDTranslation by DIN-Sprachendienst.In case of doubt, the German-language original shall be considered authoritative.10.15 DIN EN ISO 10504:2015-11 2 A comma is used as the decimal marker. National foreword This document has been prepared by Technical Committee ISO/TC 93 “Starch (inc

6、luding derivatives and by-products)” (Secretariat: BSJ, Jamaica). At present a DIN committee does not exist for this standard since the parties concerned have not shown any interest in work on the subject. The DIN Standards corresponding to the International Standards referred to in Clause 2 of this

7、 standard are as follows: ISO 3696 DIN ISO 3696 ISO 5381 DIN EN ISO 5381 Amendments This standard differs from DIN EN ISO 10504:2000-08 as follows: a) the title has been changed; b) the standard has been editorially revised; c) the Notes in subclauses 4.3 and 5.5 have been changed into footnotes and

8、 edited to make them more understandable; d) the Note in subclause 5.6 has been incorporated as normative text; e) Annex A has been updated. Previous editions DIN EN ISO 10504: 2000-08 National Annex NA (informative) Bibliography DIN ISO 3696, Water for analytical laboratory use Specification and te

9、st methods DIN EN ISO 5381, Starch hydrolysis products Determination of water content Modified Karl Fischer method EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 10504 July 2015 ICS 67.180.20 Supersedes EN ISO 10504:2000 English Version Starch derivatives - Determination of the composition

10、 of glucose syrups, fructose syrups and hydrogenated glucose syrups - Method using high-performance liquid chromatography (ISO 10504:2013) Produits drivs de lamidon - Dtermination de la composition des sirops de glucose, des sirops de fructose et des sirops de glucose hydrogns - Mthode par chromatog

11、raphie en phase liquide haute performance (ISO 10504:2013) Strkederivate - Bestimmung der Zusammensetzung von Glucosesirup, Fructosesirup und hydriertem Glucosesirup - Verfahren mittels Hochleistungs-Flssigchromatographie (ISO 10504:2013) This European Standard was approved by CEN on 19 June 2015. C

12、EN 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-date lists and bibliographical references concerning such national standards may be obtained on ap

13、plication 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 translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC

14、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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, L

15、atvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION C O M I T EUR O P EN D E NOR M A L I S ATI O N EUR O P I S C HES KOM I T E E F R NOR M UNG CEN-CENELE

16、C 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 E Contents PageEuropean foreword .31 Scope . 42 Normative references 43 Principle 44 Reagents 45 Apparatus .

17、 56 Procedure. 66.1 Choice of column . 66.2 System start-up . 66.3 Calibration of column 66.4 Sample preparation 76.5 Sample analysis 77 Calculation 78 Precision . 88.1 Repeatability . 88.2 Reproducibility . 8Annex A (informative) Examples of standard solutions 10DIN EN ISO 10504:2015-11 EN ISO 1050

18、4:2015 (E) 2 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 CEN. This European Standard shall

19、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 of this docume

20、nt 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 countries are b

21、ound 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, Poland, Portu

22、gal, 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. DIN EN ISO 10504:2015-11 EN ISO 10504:2015 (E) 3 1 ScopeThis International Standard descr

23、ibes 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, fructose, sorbitol, m

24、annitol, 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 references, only the ed

25、ition 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 Modified Karl Fische

26、r 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 electronic integra

27、tor.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-exchange resin.The wat

28、er 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 water-purificatio

29、n 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 analysed.NOTE Suitab

30、le reference materials for the constituents listed in Clause 1 can be obtained from established chemical companies.DIN EN ISO 10504:2015-11 EN ISO 10504:2015 (E) 4 4.3 Ion-exchange resins, for off-line demineralization of samples.Salts present in the sample will co-elute from the column, and will be

31、 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 following resins1):a) Cation type:1)

32、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 form;2) 200 mesh to 400 mesh in th

33、e 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, capable of maintaining the column at tem

34、peratures 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 voltage output of the detector.5.

35、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 available from several major column sup

36、pliers.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.Commercially available syrups are u

37、sually 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. Experience in several laboratories

38、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 endorsement by ISO of these products. Equiv

39、alent 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. (This information is given for the con

40、venience 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.)DIN EN ISO 10504:2015-11 EN ISO 10504:2015 (E) 5 6 Procedure6.1 Choice of columnFor general applicat

41、ions, 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 about 6 % or more. In such instances better resoluti

42、on 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 the injector to the inlet of the column (or guard c

43、olumns, 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 column according to the suppliers recommendations. Enter

44、 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 of the signal from the sample cell. Set the require

45、d 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 (tri-itol and above), no commercial standards are a

46、vailable.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 integrator response, should show a variation of 0,1 % or less

47、 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 m

48、olecular 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 as close as possible to those of the samples to be analysed. These should be prepared at the chosen concentration (see 4.2).NOTE An example is given in Annex A.6.3.4 Inject the chosen aliquot portion twice into the chromatograph. The quantity injecte