1、| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS EN 1528-3 : 1997 The E
2、uropean Standard EN 1528-3 : 1996 has the status of a British Standard ICS 67.040 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 3. Clean-up methodsBS EN 1528-3 : 1997 This British Standard, havi
3、ng 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 June 1997 BSI 1997 The following BSI references relate to the work on this standard: Committee reference AW/-/3 Draft for comm
4、ent 94/505376 DC ISBN 0 580 27381 4 Amendments issued since publication Amd. No. Date Text affected Committees responsible for this British Standard The preparation of this British Standard was entrusted to Technical Panel AW/-/3, Food analysis Horizontal methods, upon which the following bodies wer
5、e represented: Association of Public Analysts Department of Trade and Industry Food and Drink Federation Institute of Food Science and Technology Ministry of Agriculture Fisheries and Food (Laboratory of the Government Chemist) Royal Society of ChemistryBS EN 1528-3 : 1997 BSI 1997 i Contents Page C
6、ommittees responsible Inside front cover National foreword ii Foreword 2 Text of EN 1528-3 3ii BSI 1997 BS EN 1528-3 : 1997 National foreword This British Standard has been prepared by Technical Panel AW/-/3 and is the English language version of EN 1528-3: 1996 Fatty food Determination of pesticide
7、s and polychlorinated biphenyls (PCBs) Part 3 : Clean-up methods published by the European Committee for Standardization (CEN). EN 1528-3 was produced as a result of international discussions in which the United Kingdom took an active part. Cross-references Publication referred to Corresponding Brit
8、ish Standard EN 1528-1 : 1996 BS EN 1528-1 : 1997 Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 1 : General EN 1528-2 : 1996 BS EN 1528-2 : 1997 Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 2 : Extraction of fat, pesticides and P
9、CBs, and determination of fat content EN 1528-4 : 1996 BS EN 1528-4 : 1997 Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 4 : Determination, confirmatory tests, miscellaneous Compliance with a British Standard does not of itself confer immunity from legal obligation
10、s. Summary of pages This document comprises a front cover, an inside front cover, pages i and ii, the EN title page, pages 2 to 16, an inside back cover and a back cover.CEN European Committee for Standardization Comite Europe en de Normalisation Europa isches Komitee fu r Normung Central Secretaria
11、t: rue de Stassart 36, B-1050 Brussels 1996 Copyright reserved to CEN members Ref. No. EN 1528-3 : 1996 E EUROPEAN STANDARD EN 1528-3 NORME EUROPE ENNE EUROPA ISCHE NORM November 1996 ICS 67.040 Descriptors: Food products, edible fats, chemical analysis, determination of content, pesticides, polychl
12、orobiphenyl, purity, chromatography English version Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 3 : Clean-up methods Aliments gras Dosage des pesticides et des polychlorobiphe nyles (PCB) Partie 3 : Me thodes de purification Fettreiche Lebensmittel Bestimmung von
13、 pestiziden und polychlorierten Biphenylen (PCB) Teil 3 : Reinigungsverfahren This European Standard was approved by CEN on 1996-10-27. 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
14、standard without 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 oth
15、er language made 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, Denmark, Finland, France, Germany, Greece, Iceland,
16、Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.Page 2 EN 1528-3 : 1996 BSI 1997 Foreword This European Standard has been prepared by Technical Committee CEN/TC 275, Food analysis, horizontal methods, the secretariat of which is held by DIN. T
17、his 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 May 1997, and conflicting national standards shall be withdrawn at the latest by May 1997. According to the CEN/CENELEC Internal Regulations, the nat
18、ional standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. This European St
19、andard consists of the following Parts. Part 1 General presents the scope of the standard and describes general considerations with regard to reagents, apparatus, gas chromatography etc., applying to each of the analytical methods selected. Part 2 Extraction of fat, pesticides and PCBs, and determin
20、ation of fat content presents a range of analytical procedures for extracting the fat portion containing the pesticide and PCB residues from different groups of fat-containing foodstuffs. Part 3 Clean-up methods presents the details of methods A to H for the clean-up of fats and oils or the isolated
21、 fat portion, respectively, using techniques such as liquidliquid partition, adsorption or gel permeation column chromatography. Part 4 Determination, confirmatory tests, miscellaneous gives guidance on some recommended techniques for the determination of pesticides and PCBs in fatty foodstuffs and
22、on confirmatory tests, and lists a clean-up procedure for the removal of the bulk of lipids when analysing large quantities of fat. Contents Page Foreword 2 Introduction 3 1 Scope 3 2 Normative references 3 3 Principle 3 4 General 3 5 Method A: Liquidliquid partitioning with acetonitrile and clean-u
23、p on a Florisil column (AOAC) 1 4 6 Method B: Liquidliquid partitioning with dimethylformamide and clean-up on a Florisil column (Specht) 2 5 7 Method C: Column chromatography on activated Florisil (AOAC) 3 7 8 Method D: Column chromatography on partially deactivated Florisil (Stijve) 4 8 9 Method E
24、: Column chromatography on partially deactivated aluminium oxide (Greve & Grevenstuk) 5 10 10 Method F: Gel permeation chromatography (GPC) (AOAC) 6 11 11 Method G: Gel permeation chromatography (GPC) and column chromatography on partially deactivated silica gel (Specht) 7 12 12 Method H: High perfo
25、rmance gel permeation chromatography (HPGPC) (MAFF) 8 13 Annexes A (informative) Purification of some solvents 15 B (informative) Bibliography 15Page 3 EN 1528-3 : 1996 BSI 1997 1) Florisil is an example of a suitable product available commercially. This information is given for the convenience of u
26、sers of this European Standard and does not constitute an endorsement by CEN of the product named. Introduction This European Standard comprises a range of multi-residue methods of equal status: no single method can be identified as the prime method because, in this field, methods are continuously d
27、eveloping. The methods selected for inclusion in this standard have been validated and are widely used throughout Europe. Any variation in the methods used should be shown to give acceptable results. The residues to be analysed in this European Standard are associated with the fat portion of the sam
28、ples. In addition to the residues, the extracts obtained in accordance with EN 1528-2 : 1996 or in accordance with the following methods contain material including fats and other lipids, which could interfere in the analysis. To purify the crude extracts or the fats and oils to be analysed, several
29、methods may be used. This European Standard contains the following clean-up methods that have been subjected to interlaboratory studies and are adopted throughout Europe. Method A. Liquidliquid partitioning with acetonitrile and clean-up on a Florisil column (AOAC) 1. Method B. Liquidliquid partitio
30、ning with dimethylformamide and clean-up on a Florisil column (Specht) 2. Method C. Column chromatography on activated Florisil (AOAC) 3. Method D. Column chromatography on partially deactivated Florisil (Stijve) 4. Method E. Column chromatography on partially deactivated aluminium oxide (Greve & Gr
31、evenstuk) 5. Method F. Gel permeation chromatography (GPC) (AOAC) 6. Method G. Gel permeation chromatography (GPC) and column chromatography on partially deactivated silica gel (Specht) 7. Method H. High performance gel permeation chromatography (HPGPC) (MAFF) 8. 1 Scope This Part of EN 1528 specifi
32、es the details of methods A to H for the clean-up of fats and oils or the isolated fat portion, respectively, using techniques such as liquidliquid partition, adsorption or gel permeation column chromatography. The applicable usage of the methods A to H is given in detail in each method described. N
33、OTE. See also EN 1528-4 which lists a clean-up procedure for the removal of the bulk of lipids when analysing large quantities of fat. 2 Normative references This European Standard incorporates by dated or undated reference, provisions from other publications. These normative references are cited at
34、 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 revision. For undated references the latest edition of
35、the publication referred to applies. EN 1528-1 : 1996 Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 1 : General EN 1528-2 : 1996 Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 2 : Extraction of fat, pesticides and PCBs, and determi
36、nation of fat content EN 1528-4 : 1996 Fatty food Determination of pesticides and polychlorinated biphenyls (PCBs) Part 4 : Determination, confirmatory tests, miscellaneous 3 Principle Removal of interfering materials from the sample extract to obtain a solution of the extracted residue in a solvent
37、 which is suitable for quantitative examination by the selected method of determination. 4 General Each liquidliquid partition shall be performed in a separating funnel with a 2 min shake, with occasional release of the pressure by opening the stopcock, with the funnel inverted. If vigorous shaking
38、produces very stable emulsions, gentle shaking for longer periods may be preferable. Emulsions may be broken by adding 1 ml to 2 ml of saturated sodium chloride or sodium sulfate solution, by warming under the hot water tap or by centrifuging. When separating layers, leave any emulsified interface w
39、ith the portion to be re-extracted or discarded. The activity of the Florisil used for column chromatography in this standard shall be checked at regular intervals and, if necessary, shall be properly adjusted as described in the methods concerned. NOTE 1. Florisil 1) is widely used as an adsorbent
40、in residue analysis because of its high capacity for binding lipids. Its activity, however, can vary considerably, depending on the manufacturing batch and on the shipping and storage conditions.Page 4 EN 1528-3 : 1996 BSI 1997 2) 1, 2, 3, 4, 10, 109-Hexachloro-1, 4, 49,5,8,89 -hexahydro-1,4-endo-5,
41、8-endodimethanonaphthalene. The rate of elution of chromatographic columns is usually specified but should generally be in the range of 1 ml/min to 5 ml/min. In the analysis of organochlorine pesticides, at this stage of the procedure, the addition of a known quantity of the rather volatile pentachl
42、orobenzene (or 1,7-dibromoheptane) and a less volatile indicator compound (e.g. 1, 2, 3, 4-tetrachloronaphthalene or isodrin 2) ) is recommended. Use pentachlorobenzene as an indicator of possible losses of pesticides during the evaporation stage by comparison of its peak area (height) with the peak
43、 area (height) of the less volatile indicator compound. An added indicator compound can also be used as an internal standard material for identification (relative retention time) and quantification purposes. With the electron capture detector (ECD), however, coextractives may give peaks (positive an
44、d negative) emerging with the same retention time as the internal standard material. Unless otherwise specified, evaporation of organic solvent solutions should not be allowed to go to complete dryness as this can result in compound losses. NOTE 2. A bibliography relating to the methods is included
45、(see annex B). 5 Method A. Liquidliquid partitioning with acetonitrile and clean-up on a Florisil column (AOAC) 1 5.1 Applicability This method has been shown to be applicable for the determination of 17 organochlorine pesticides and metabolites, the PCB indicator congeners and 6 organophosphorus pe
46、sticides (in special cases), as given in annex A of EN 1528-1 : 1996. 5.2 Principle Extraction of the residues, together with the fat, from the sample by one of the procedures described in EN 1528-2 : 1996. Concentration of the extract almost to dryness, redissolving in light petroleum and partition
47、ing of the residues into acetonitrile. After diluting the acetonitrile with an excess of water, partitioning of the residues into light petroleum. Chromatography of the concentrated organic phase on a Florisil column using light petroleum/diethyl ether for elution. Concentration of the eluates for e
48、xamination by GC. 5.3 Reagents and materials All reagents and materials used shall be suitable for the analysis of residues of pesticides and PCBs and shall be in accordance with clause 4 of EN 1528-1 : 1996. If purification is necessary, the procedures given in annex A are appropriate. 5.3.1 Aceton
49、itrile. 5.3.2 Light petroleum, having a boiling range from 40 C to 60 C. 5.3.3 Acetonitrile (5.3.1), saturated with light petroleum (5.3.2). 5.3.4 Diethyl ether, peroxide free. Distil and stabilize with 2,0 % of its volume with absolute ethanol. 5.3.5 Eluting mixture A, light petroleum (5.3.2) and diethyl ether (5.3.4) 9 4:6( V/V). 5.3.6 Eluting mixture B, light petroleum (5.3.2) and diethyl ether (5.3.4)8 5:1 5( V/V). 5.3.7 Eluting mixtur
