EN 12630-1999 en Fruit and Vegetable Juices - Determination of Glucose Fructose Sorbitol and Sucrose Contents - Method Using High-Performance Liquid Chromatography《水果和蔬菜汁 葡萄糖 果糖 山梨.pdf

1、 STD-BSI BS EN 12b30-ENGL 1999 Lb24669 0790900 70b BRITISH STANDARD Fruit and vegetable juices - Determination of glucose, fructose, sorbitol and sucrose contents - Method using high-performance liquid chromatography The European Stanckd EN 12630:1999 has the status of a British standard ICs 67.160.

2、20 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW 3s EN 12630:1999 BS EN 126301999 been prepared under the dimtion of the Consumer AmdNo. Date Products and Services Sector committee, was published under the authority of the Sandars Committee and comes into effect on 15 August

3、 1999 0 BSI 081999 ISBN O 580 32213 O National foreword Comments This British Standard is the English language version of EN 1263OA999. The TJK participation in its preparation was entrusted to Technicd Committee AWfil, Fruit and vegetables juices, which has the responsibility to: - aid enquirers to

4、 understand the text; - present to the responsible European committee any enquiries on the - monitor related international and European developments and promulgate interpretation, or proposals for change, and keep the UK inter Sorbit- und k- Mthode par chromatographie liquide haute performance Hochl

5、flWMes Verfahren This European Standard was approved by CEN on 8 January 1999. CEN members are bound to compiy with the CENKENELEC intemai Regulations which stipuiak the conditions for giving this European Stanard the status of a national standard without any alteration. Upto-date lists and bibhogra

6、phid 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 officiai versions (English, French, German). A version in any other language made by tmnsMon under the responsibii of a CEN member

7、 into its own language and notified to the Centmi Secreariat has the same status as the officiai versions. CEN members are the national standards bodies of Ausria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Im, Luxembourg, Netherlands, Norway, Portugai, Spa

8、in, Sweden, Switzerland and United Kingdom. European Committee for Stanmon Comit Europen de Normahation Europisches Komitee fr Normung Central Secretariat: rue de Stassart 36, B-1060 Brussels O 1999 CEN AU rights of exploitation in any form and by any means reserved worldwide for CEN national Member

9、s. Ref. No. EN 126301999 E STDOBSI BS EN 32630-ENGL 3999 W Lb24bb9 O790903 435 Page 2 EN l263oam Foreword This European standard has been prepared by Whnical Committee CENiIY: 174, Fruit and vegetable juices - Methods of anaiysii, the Secretariat of which is held by AFNOR. This European standard sha

10、ll be given the status of a nationai standard, either by publication of an identical text or by endomement, at the latRst by August 1999, and conicting national standards shall be withdrawn at the latest by August 1999. According to the CENKENELEC internal Regulations, the national standards organht

11、ions of the following countries are bound to implement this European Stan* Austsia, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, I-, Luxembourg, Netherlands, Norwax Portugal, Spain, Sweden, Switzerland and the United Kingdom. Contents Foreword 1 Scope 2 Norma

12、tive references 3 Symbols and abbreviations 4 Principle 5 Reagents 6 APP- 7 Procedure 8 Calculation 9 Precision 10 Testreport Annex A (informative) Bibliography Annex B (informative) Statistical results of the interiabomry test Annex C (informative) Example chromatogram for the determination of suga

13、rs and sorbitol by HPI Page 2 3 3 3 3 3 3 3 4 4 5 6 6 8 Q BSI O-1999 STD-BSI BS EN 12630-ENGL 1979 E Lb24669 0770904 351 Page 3 EN 1263 NOTE The rotational frequency required to give correct centrifugai acceleration can be calculated from the following equation: g acceleration due to gravity at the

14、surface of the a=11,18XX (1 - :o07 where earth (9,81 m/sz). 3.2 Abbreviations a is the centrifugai acceleration; For the purposes of this standard, the following abbreviations apply: T is the radius of the centrifuge in centimetres, measured from the midpoint (the centrifuge axis) to the bottom of t

15、he centrifuge tube when swung outwards; n is the rotational frequency per minute. HPLC High-performance liquid chromatography; EDTA Ethylenediaminektm-acetic acid 6.6 Centnfuge tubes. 4 Principle 7 Procedure The sugars and sorbitol are separated on a 7.1 General Analysii by this method shaU be on a

16、volumetric basis, the results being expressed per litre of sample. The analysis of concen-d products may also be carried relative density in thic case, the relative density shall be indicated Based on a weighed sample and taking the dilution factor for analvsis into account., the results cationexcha

17、nge resin (sulfonated poiystyrene ivinylbenzene copolymer in the Caz+ form) by isomtic elution, using an aqueous solution of caicium sohito1 are detected using a differential refractive index detector and quantified using the extend standard method. disodium-EDTA as the mobile phase* The out on a vo

18、lume is the dilution fador (5 for juices; for diluted concentrates the factor shaii be calculated); particuar sugar or sorbitol, and is calculated using the foilowing equation: (2) p=- XF where P F RF is the appropriate response factor for the (3) Report the concentxations of glucose, fructose, sorb

19、itol and sucrose in grams per itre to one decimai place. NOTE 1 Separate response factors should be determined for giucose, fructose, sorbitol and sucrose, using the equation above and the appropriate data NOTE 2 When this method was assessed in a coiiaborative study, the concentrations of the sugar

20、s were also assessed using enzymatic procedures. It was found that the enzymatic methods gave a better reproducibility for glucose and fructose, whereas the HPLC method gave a better reproducibility for sucrose. 9 Precision Details of the interlaboratory test on precision of the method are summarize

21、d in annex B. The values derived from the interlaboratory test may not be applicable to amlyse conceniration ranges and matrices other than those given in annex B. 9.1 Repeatability The absolute difference between two single results found on identical test matehl by one operator using the me apparat

22、us within the shortest feasible time intervai wiil exceed the repeaability limit r in not more than 5 % of the cases. The dues are given in Table 1. Table 1 Table 2 Grape juice 5,l nm i) nm = not measured. as not Dresent in this hice. Ps is the peak area or peak height of the particuiar sugar or sor

23、bitol in the standard Chromatogram; ps is the mass concentmtion of the particular sugar or sorbitol in the standard solution. O BSI 081999 STD-BSI BS EN 12b30-ENGL 1999 Lb24669 079090b 124 Page 6 EN lrn - a reference to this European standard; - the date and type of sampiing proceure (i known); - th

24、e date of receipt; -thedateoftest; - test resuits and the units in which they have been expressed; - whether the repeatability of the method has been verified; - any particular points observed in the com of the test; - any operations not specifed in the method, or regarded as optional, which might h

25、ave affected the results. O BSI 08-1999 STD-BSI BS EN 22b30-ENGL 2999 m Lb24bb9 OO9O Ob0 = Number of laboratories retained after ehhating outliers Number of outiers (laboratories) Page 6 EN um1999 A B 9 10 2 1 Annex A (informative) Bibliography IS0 57251986, Pmision of test methods - Detemzinatwn of

26、 repeatability and rejmduciaity for a standard test method by inter- tests. Determination of glucose,fmcbse, SuCTose and sorbitol by HPLC: No. 67,1995. In: Analyses Colection/nteona,l Federation of Fniit Juice Producers. Loose-leaf edition, as of 1995. Zug: Swiss Fruit UNon Repeatability standard de

27、viation, Sr (l) Repeatability reMve standard deviation, RSDr (%) Repeatability limit, r (gli) Reproducibility standard deviation, Q i) Annex B (idormative) Statistical results of the interlaboratory test in accordance with Is0 57251986, the foilowing parameters have been defined in an interlabomtory

28、 test. (For itemlme on the method, see annex A). The test was conducted by the Max von Pettenkofer Institute of the Federai Heaith Office, Food Chemistry Department, Berlin, BRD - year of interiabomtoy test: 1993; - number of laboratores: 11; - number of samples: 4. nble B.l- Glucose Samule o746 0,s

29、 198 196 43 1,1 1,Ol 1,m Reproducibility reiative standard deviation, RSh (%) Reproducibility limit, R (g/i) Number of accepted results I29 I32 Mean value. Y dl 125.2 124.9 490 4,4 299 371 Number of laboratories retained after elhinahg outhers Number of outliers (laboratories) Number of accerked res

30、ults A B 10 11 1 O 31 34 29 29 31.6 77.0 Mean due, Z (gA) Repeatabiity standard deviation, Sr ) 0,49 I 0771 I 62,3 27,7 27,3 78,7 4% 09% 0741 0,60 2,18 1,79 Repeatability reiative standard deviation, RSDr (%) Repeaabiity limit, r (gA) Reproducibility standard deviation, Q (g4) D Grapejuice 0,9 194 1

31、95 0,s 196 171 192 47 2,16 1,ll 2,29 3,17 Table B.2 - Fructose Reproducibiity relative standard deviation, RSh (%) Reproducibility limit, R (gli) I Samule I 3,5 470 894 490 631 3J 695 970 34 31 O BSI 081999 STD-BSI BS EN L2b30-ENGL 3999 = Lb24bb9 0790908 TT7 Number of laboratories retained after eli

32、minating outliers I Number or outiiem (laboratories) Number of accepted resuits Mean due, 5 (gh) Repeatability standard deviation, s, (gA) Repeatability reMve standard deviation, RSD, (%) Repeatability limic r (gl) Reproducibility standard deviation, SR (gA) Reproducibility relative standard deviati

33、on, Rsh (%) Reproducibility limit, R (g4) Page 7 EN 1263hese and other benefits contact Membership Admhkmtion. Tek 020 8996 7002. Fax: 020 8996 7001. Copyright Zopynght subsists in all BSI publications. BSI also holds the copyright, in the UK, of the publications of the international standardization

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35、he free use, in the course of implementing the standard, of necessary details such as symbols, and size, type or grade designations. If these details are to be used for any other purpose than implementation then the prior written permission of BSI must be obtained if permission is granted, the terms may include royalty payments or a licensing agreement. Details and advice can be obtained from the Copyright Manager. Tel O20 8996 7070. Previous page is blank