1、BSI Standards PublicationBS EN ISO 15753:2016Animal and vegetable fatsand oils Determinationof polycyclic aromatichydrocarbons (ISO 15753:2016)BS EN ISO 15753:2016 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN ISO15753:2016. It supersedes BS EN ISO 15753:2006+
2、A1:2011 which iswithdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee AW/307, Oilseeds, animal and vegetable fats and oils andtheir by-products.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not p
3、urport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2016. Published by BSI StandardsLimited 2016ISBN 978 0 580 91529 1ICS 67.200.10Compliance with a British Standard cannot confer immunity fromlegal obligati
4、ons.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 30 April 2016.Amendments issued since publicationDate Text affectedEUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN ISO 15753 April 2016 ICS 67.200.10 Supersedes EN ISO 15753:2006English
5、Version Animal and vegetable fats and oils - Determination of polycyclic aromatic hydrocarbons (ISO 15753:2016) Corps gras dorigines animale et vgtale - Dtermination des hydrocarbures aromatiques polycycliques (ISO 15753:2016) Tierische und pflanzliche Fette und le - Bestimmung von polycyclischen ar
6、omatischen Kohlenwasserstoffen (ISO 15753:2016) This European Standard was approved by CEN on 11 March 2016. 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 any altera
7、tion. 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 language made
8、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, Estonia
9、, 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 STANDAR
10、DIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2016 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 15753:2016 EBS EN ISO 15753:2016EN ISO
11、 15753:2016 (E) 3 European foreword This document (EN ISO 15753:2016) has been prepared by Technical Committee ISO/TC 34 “Food products” in collaboration with Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products - Methods of sampling and analysis” the se
12、cretariat of which is held 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 October 2016, and conflicting national standards shall be withdrawn at the latest by October 2016. Attention is
13、 drawn to the possibility that some of the elements 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 15753:2006. According to the CEN-CENELEC Internal Regulations, the
14、 national standards organizations of the following countries 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
15、, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 15753:2016 has been approved by CEN as EN ISO 15753:2016 without any modification. BS EN ISO 15753:2016ISO
16、 15753:2016(E)Foreword iv1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 25 Reagents and materials . 26 Apparatus . 37 Sampling 48 Sample preparation . 59 Procedure for determination of PAHs from fats and oils: General method 59.1 Preliminary remarks 59.2 Blank 59.3 Determi
17、nation of recovery values (without matrix) 59.4 Extraction (liquid/liquid extraction) . 59.5 Purification on C18-bonded phase cartridge (solid/liquid extraction). 69.6 Purification on Florisil-bonded phase cartridge (solid/liquid extraction). 710 Procedure for determination of PAHs from fats and oil
18、s: Method specific for coconut oil . 810.1 First extraction (liquid/liquid extraction) 810.2 Second extraction (liquid/liquid extraction) 810.3 Purification on C18-bonded phase cartridge (solid/liquid extraction). 810.4 Purification on Florisil-bonded phase cartridge (solid/liquid extraction). 911 H
19、igh-performance liquid chromatography (HPLC) .1011.1 Operating conditions 1011.2 Detection parameters . 1011.3 Analysis of samples and standards . 1211.4 Confirmation of the presence of PAHs . 1212 Expression of results .1213 Precision 1313.1 Interlaboratory test 1313.2 Repeatability 1313.3 Reproduc
20、ibility 1314 Test report 13Annex A (informative) Recovery values, flow charts, chromatograms and injection sequences .15Annex B (informative) Results of the interlaboratory test .20Bibliography .23 ISO 2016 All rights reserved iiiContents PageBS EN ISO 15753:2016ISO 15753:2016(E)ForewordISO (the Int
21、ernational Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee ha
22、s 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 collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical
23、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 approval criteria needed for the different types of ISO documents should be noted. This document was drafted in acc
24、ordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Detai
25、ls 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 (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an end
26、orsement.For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee respo
27、nsible for this document is ISO/TC 34, Food products, Subcommittee SC 11, Animal and vegetable fats and oils.This second edition cancels and replaces the first edition (ISO 15753:2006), of which it constitutes a minor revision. It also incorporates Amendment ISO 15753:2006/Amd.1:2011. A non-applicab
28、ility statement for milk and milk products has been added to the Scope.iv ISO 2016 All rights reservedBS EN ISO 15753:2016INTERNATIONAL STANDARD ISO 15753:2016(E)Animal and vegetable fats and oils Determination of polycyclic aromatic hydrocarbons1 ScopeThis International Standard describes two metho
29、ds for the determination of 15 polycyclic aromatic hydrocarbons (PAHs) in animal and vegetable fats and oils: a general method; a specific method for coconut oil and vegetable oils with short-chain fatty acids.These methods are not quantitative for the very volatile compounds such as naphthalene, ac
30、enaphthene and fluorene. Due to interferences provided by the matrix itself, palm oil and olive pomace oil cannot be analysed using this method.The quantification limit is 0,2 g/kg for almost all compounds analysed, except for fluoranthene and benzo(g,h,i)perylene, where the quantification limit is
31、0,3 g/kg, and indeno(1,2,3-c,d)pyrene, where the quantification limit is 1,0 g/kg.NOTE The results for olive pomace oil in Annex B show that this method is not applicable to this type of oil. The precision data determined are very poor. Milk and milk products (or fat coming from milk and milk produc
32、ts) are excluded from the scope of this International Standard.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 edition cited applies. For undated references, the
33、latest edition of the referenced document (including any amendments) applies.ISO 661, Animal and vegetable fats and oils Preparation of test sample3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1polycyclic aromatic hydrocarbonPAHcompound that co
34、ntains two or more condensed (fused) aromatic hydrocarbon rings and the content of which can be determined according to the method specified in this International StandardNote 1 to entry: The content is given in micrograms per kilogram.Note 2 to entry: In general, PAHs are divided into light PAHs wi
35、th two to four aromatic rings, and heavy PAHs with five or more aromatic rings.EXAMPLE Light PAHs include:naphthalene (CAS RN 91-20-3), acenaphthene (CAS RN 83-32-9), acenaphthylene (CAS RN 208-96-8), fluorene (CAS RN 86-73-7), anthracene (CAS RN 120-12-7), phenanthrene (CAS RN 85-01-8), fluoranthen
36、e (CAS RN 206-44-0), chrysene (CAS RN 218-01-9), benz(a)anthracene (CAS RN 56-55-3), pyrene (CAS RN 129-00-0). ISO 2016 All rights reserved 1BS EN ISO 15753:2016ISO 15753:2016(E)Heavy PAHs include:benzo(a)pyrene (CAS RN 50-32-8), benzo(b)fluoranthene (CAS RN 205-99-2), benzo(k)fluoranthene (CAS RN 2
37、07-08-9), benzo(g,h,i)perylene (CAS RN 191-24-2), dibenz(a,h)anthracene (CAS RN 53-70-3), indeno(1,2,3-c,d)pyrene (CAS RN 193-39-5).4 PrincipleThe polycyclic aromatic hydrocarbons are extracted with an acetonitrile/acetone mixture followed by purification on C18 reversed-phase and then Florisil bond
38、ed-phase cartridges. Determination of the content of the individual polycyclic aromatic hydrocarbons after separation is achieved by means of high-pressure liquid chromatography (HPLC) and by measuring the fluorescence at various excitation and emission wavelengths.5 Reagents and materialsWARNING At
39、tention is drawn to the regulations governing the handling of dangerous matter. Technical, organizational and personal safety measures should be followed.Use only reagents of recognized analytical grade unless otherwise stated.Check the quality of solvents before use by concentrating the solvent abo
40、ut 1 000 times by evaporation and analysing the concentrate by HPLC (300 ml to 300 l). The chromatogram shall be free from peaks in the elution area of PAHs.5.1 Methanol, “ultra resi-analysed” grade.1)5.2 Hexane, HPLC grade.1)5.3 Acetonitrile, HPLC grade.1)5.4 Acetone, HPLC grade.1)5.5 Dichlorometha
41、ne, HPLC grade.1)5.6 Toluene, HPLC grade.1)5.7 Water, HPLC grade.1)5.8 Tetrahydrofuran, HPLC grade.1)5.9 Solvent mixture 1: acetonitrile/acetone (volume fraction 60 %/40 %).Quantity used per sample: 41 ml for general method, 36 ml for specific method for coconut oil.5.10 Solvent mixture 2: acetonitr
42、ile/acetone (volume fraction 80 %/20 %).Quantity used per sample: 2 11 ml for method specific for coconut oil.5.11 Solvent mixture 3: hexane/dichloromethane (volume fraction 75 %/25 %).Quantity used per sample: 7 ml for general method, 2 7 ml for method specific for coconut oil.1) These can be obtai
43、ned from, for example, Baker. The information given in the footnotes is for the convenience of users of this document 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 2016 All rights reservedBS EN
44、ISO 15753:2016ISO 15753:2016(E)5.12 Mixture of tetrahydrofuran/methanol (volume fraction 50 %/50 %).5.13 Standard solution with 16 certified EPA Priority PAHs in toluene,2)at a concentration of 100 g/ml (100 mg/l): naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluora
45、nthene, pyrene, benz(a)anthracene, chrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene, dibenz(a,h)anthracene, benzo(g,h,i)perylene, indeno(1,2,3-c,d)pyrene.NOTE 1 This is stored at -20 C.Before use, allow the solution to warm up to ambient temperature for at least 1 h.NOTE 2 Acenap
46、hthylene is not fluorescent and, thus, it cannot be determined by these methods.5.14 Stock standard solution, 200 ng/ml (200 g/l).Add 100 l of standard solution (5.13) with a 250 l syringe (6.11) to a 50 ml volumetric flask (6.20) and dilute to the mark with acetonitrile.5.15 Working standard soluti
47、on, 50 ng/ml (50 g/l).Add 250 l of stock standard solution (5.14) with a 250 l syringe (6.11) to 750 l of THF/methanol mixture (5.12) or acetonitrile (5.3).5.16 C18 bonded-phase cartridges,3)2 g phase, 12 ml capacity.5.17 Florisil bonded-phase cartridges,3)500 mg phase, 3 ml capacity.5.18 Stream of
48、nitrogen, pressure regulated at 34,5 kPa (5 psi, about 1,5 l/min).6 ApparatusUsual laboratory apparatus and, in particular, the following.The use of disposable glass tubes is acceptable. The general use of glass is necessary as plastics can contain PAHs.6.1 Centrifuge, capable of attaining at least
49、4 000 min1, suitable for 100 ml and 10 ml tubes.6.2 HPLC system with binary gradient elution, with solvent reservoir of 1 l capacity, a mobile phase liner filter, pump, autosampler, column temperature regulation set at 25 C, fluorescence detector programmable over time for various excitation and emission wavelengths, and computer-assisted acquisition and data treatment.6.3 C18 reversed-phase column,4)250 mm in length, 4,6 mm internal diameter, 5 m particles, suitable for PAH analysis.6.4 Vortex mixer.2
