1、| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS EN 12137 : 1998 The Eu
2、ropean Standard EN 12137 : 1997 has the status of a British Standard ICS 67.160.20 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Fruit and vegetable juices Determination of tartaric acid in grape juices Method by high performance liquid chromatographyThis British Standard, h
3、aving been prepared under the direction of the Consumer Products and Services Sector Board, was published under the authority of the Standards Board and comes into effect on 15 February 1998 BSI 1998 ISBN 0 580 28960 5 BS EN 12137 : 1998 Amendments issued since publication Amd. No. Date Text affecte
4、d National foreword This British Standard is the English language version of EN 12137 : 1997. The UK participation in its preparation was entrusted to Technical Committee AW/21, Fruit and vegetable juices, which has the responsibility to: aid enquirers to understand the text; present to the responsi
5、ble European committee any enquiries on the interpretation, or proposals for change, and keep the UK 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 secr
6、etary. ISO 5725 : 1986, to which informative reference is made in the text, has been superseded by ISO 5725-1 : 1994, ISO 5725-2 : 1994, ISO 5725-3 : 1994, ISO 5725-4 : 1994 and ISO 5725-6 : 1994, which are identical with BS ISO 5725 Accuracy (trueness and precision) of measurement methods and resul
7、ts, BS ISO 5725-1 : 1994 General principles and definitions, BS ISO 5725-2 : 1994 Basic method for the determination of repeatability and reproducibility of a standard measurement method, BS ISO 5725-3 : 1994 Intermediate measures of the precision of a standard measurement method, and BS ISO 5725-6
8、: 1994 Use in practice of accuracy values. Cross-references The British Standards which implement international or European publications 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 F
9、ind facility of the BSI Standards Electronic Catalogue. 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 title page, pages 2 to 6, an inside back cover and a back cover.
10、CEN European Committee for Standardization Comite Europe en de Normalisation Europa isches Komitee fu r Normung Central Secretariat: rue de Stassart 36, B-1050 Brussels 1997 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 12137 : 1
11、997 E EUROPEAN STANDARD EN 12137 NORME EUROPE ENNE EUROPA ISCHE NORM September 1997 ICS 67.160.20 Descriptors: Fruit and vegetable juices, grapes, chemical analysis, determination of content, tartaric acid, high performance liquid chromatography, procedure English version Fruit and vegetable juices
12、Determination of tartaric acid in grape juices Method by high performance liquid chromatography Jus de fruits et de le gumes Dosage de lacide tartrique dans les jus de raisin Me thode par chromatographie liquide a haute performance Frucht- und Gemu sesa fte Bestimmung von Weinsa ure in Traubensaft H
13、ochleistungs-flu ssigkeitschromatographisches Verfahren This European Standard was approved by CEN on 6 September 1997. 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
14、 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, German). A version in any other language made
15、 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, Finland, France, Germany, Greece, Iceland,
16、Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.Page 2 EN 12137 : 1997 BSI 1998 Foreword This European Standard has been prepared by Technical Committee CEN/TC 174, Fruit and vegetable juices Methods of analysis, the secretariat of which is he
17、ld 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 March 1998, and conflicting national standards shall be withdrawn at the latest by March 1998. According to the CEN/CENELEC Internal Re
18、gulations, 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, Portugal, Spain, Sweden, Switzerland and th
19、e United Kingdom. Contents Page Foreword 2 1 Scope 3 2 Normative references 3 3 Symbols 3 4 Principle 3 5 Reagents 3 6 Apparatus 3 7 Procedure 3 8 Calculation 4 9 Precision 4 10 Test report 4 Annexes A (informative) Bibliography 5 B (informative) Statistical results of the inter-laboratory test 5Pag
20、e 3 EN 12137 : 1997 BSI 1998 1 Scope This European Standard specifies a method for the determination of tartaric acid in grape juices by high performance liquid chromatography (HPLC). 2 Normative references This European Standard incorporates by dated or undated reference, provisions from other publ
21、ications. These normative references are cited at the appropriate places in the text and the publications are listed hereafter. For dated references, subsequent amendments to or revisions of any of these publications apply to this European Standard only when incorporated in it by amendment or revisi
22、on. For undated references the latest edition of the publication referred to applies. EN ISO 3696 : 1995 Water for analytical laboratory use Specification and test methods (ISO 3696 : 1987) ISO 5725 : 1986 Precision of test methods Determination of repeatability and reproducibility for a standard te
23、st method by inter-laboratory tests 3 Symbols For the purposes of this standard, the following symbol applies: c substance concentration 4 Principle The content of tartaric acid in grape juices is determined by HPLC (using ultraviolet detection). The separation takes place on an ion-exclusion column
24、. 5 Reagents 5.1 General Use only reagents of recognized analytical grade and only water in accordance with at least grade 1 of EN ISO 3696 : 1995. 5.2 Sulfuric acid, c(H 2 SO 4 ) = 0,005 mol/l. 5.3 Standard tartaric acid solution (C 4 H 6 O 6 ) A defined quantity of tartaric acid is dissolved in wa
25、ter. The concentration should be approximately 500 mg/l. 6 Apparatus Usual laboratory apparatus and, in particular, the following. 6.1 High performance liquid chromatograph 6.2 Ultraviolet (UV) detector, capable of measuring at a wavelength of 210 nm. 6.3 Column oven, capable of maintaining a temper
26、ature of 408C. 6.4 Separation column, ion-exclusion column made of sulfonated divinyl benzene-styrene copolymer in hydrogen form, typical particle size 10mm, (300 mm3 7,8 mm) with a cation H + precolumn. 6.5 Membrane filter, of pore size 0,45mm. 7 Procedure 7.1 Preparation of the test sample Normall
27、y products shall not be pre-treated, however dilution may be necessary and their analysis by this method shall be on a volumetric basis, results being expressed per litre of sample. The analysis of concentrated products may also be carried out on a volumetric basis, after dilution to a known relativ
28、e density. In this case the relative density shall be indicated. Based on a weighed sample and taking the dilution factor for analysis into account, the results may also be expressed per kilogram of product. In products with a high viscosity and/or a very high content of cells (for example pulp), a
29、determination on the basis of a weighed test sample is the usual procedure. Dilute grape juices at 1 to 20 volumes (concentrates 1 to 100) and directly use them for HPLC analysis after filtration through a membrane filter (6.5). (When frozen samples are used, ensure that there is no sediment in the
30、sample before dilution.) 7.2 Test procedure 7.2.1 Preparation of calibration solutions Prepare the calibration solutions in the range of 100 mg/l to 500 mg/l of tartaric acid using suitable solutions of the standard tartaric acid solution (5.3). Use the solutions as described in 7.2.2. 7.2.2 HPLC an
31、alysis Inject the calibration solutions (7.2.1) and the samples (7.1) in an HPLC system (6.1, 6.2, 6.3) with the following conditions : Eluent: 0,005 mol/l sulfuric acid (5.2); Column: separation column (6.4); Flow: e.g. 0,6 ml/min (to avoid a high pressure the flow should be increased slowly from 0
32、,2 ml/min to 0,6 ml/min during equilibration); Wavelength: 210 nm; Injection volume: typically 15ml; Running time: 20 min; Retention time: approximately 10 min for tartaric acid; Temperature: 408C.Page 4 EN 12137 : 1997 BSI 1998 8 Calculation Calculate the content of tartaric acid in the sample usin
33、g the calibration curve based on peak height. Take into account the dilution factor and the relation of the value to mass or volume. If a concentrated product has been diluted to single strength, report the relative density of the single strength sample. Report the tartaric acid concentration in gra
34、ms per litre to one decimal place. 9 Precision Details of the inter-laboratory test on the precision of the method are summarized in annex B. The values derived from the inter-laboratory test may not be applicable to analyte concentration ranges and matrices other than given in annex B. 9.1 Repeatab
35、ility The absolute difference between two single test results found on identical test material by one operator using the same apparatus within the shortest feasible time interval will exceed the repeatability limit r in not more than 5 % of the cases. The value is: r = 0,14 g/l. 9.2 Reproducibility
36、The absolute difference between two single test results on identical test material reported by two laboratories will exceed the reproducibility limit R in not more than 5 % of the cases. The value is: R = 0,33 g/l. 10 Test report The test report shall contain the following data: all information nece
37、ssary for the identification of the sample (kind of sample, origin of sample, designation); a reference to this European Standard; the date and type of sampling procedure (if known); the date of receipt; the date of test; the test results and units in which they have been expressed; whether the repe
38、atability of the method has been verified; any particular points observed in the course of the test; any operations not specified in the method or regarded as optional, which might have affected the results.Page 5 EN 12137 : 1997 BSI 1998 Annex A (informative) Bibliography Determination of tartaric
39、acid in grape juices by HPLC : No 65, 1995. In: The collected Analyses of the International Federation of Fruit Juice Producers. Looseleaf edition, 1996, Zug: Swiss Fruit Union. Annex B (informative) Statistical results of the inter-laboratory test In accordance with ISO 5725 : 1986, the following p
40、arameters have been defined in an inter-laboratory test. (For literature pertaining to the method see annex A). The test was conducted by the International Fruit Union, Paris, France. Year of inter-laboratory test 1992 Number of laboratories 11 Number of samples 4 Table B.1 Sample ABCD Number of lab
41、oratories retained after eliminating outliers 8 10 9 9 Number of outliers (laboratories) 3122 Number of accepted results 41 54 46 46 Mean value, x, (g/l) 4,19 2,55 3,77 2,15 Repeatability standard deviation, s r , (g/l) 0,0621 0,0513 0,0566 0,0260 Repeatability relative standard deviation, RSD r , (
42、%) 1,48 2,01 1,50 1,21 Repeatability limit, r, (g/l) 0,17 0,14 0,16 0,07 Reproducibility standard deviation, s R , (g/l) 0,1148 0,1253 0,1363 0,0931 Reproducibility relative standard deviation, RSD R , (%) 2,74 4,91 3,62 4,33 Reproducibility limit, R, (g/l) 0,32 0,35 0,38 0,26 Sample types: A grape
43、juice, white; B grape juice, white; C grape juice, red; D grape juice, red.BSI 389 Chiswick High Road London W4 4AL | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
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