《ISO-15753-2006.pdf》由会员分享，可在线阅读，更多相关《ISO-15753-2006.pdf（28页珍藏版）》请在三一文库上搜索。
1、 Reference number ISO 15753:2006(E) ISO 2006 INTERNATIONAL STANDARD ISO 15753 First edition 2006-09-01 Animal and vegetable fats and oils Determination of polycyclic aromatic hydrocarbons Corps gras dorigines animale et vgtale Dtermination des hydrocarbures aromatiques polycycliques Copyright Intern
2、ational Organization for Standardization Provided by IHS under license with ISO Licensee=Hong Kong Polytechnic Univ/9976803100 Not for Resale, 04/17/2007 09:44:56 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15753:2006(E) PDF disclaimer This PDF file may contain embe
3、dded typefaces. In accordance with Adobes licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In downloading this file, parties accept therein the responsibility of
4、not infringing Adobes licensing policy. The ISO Central Secretariat accepts no liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation parameters
5、were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below. ISO 2006 All rights reserved. Unless otherwise spec
6、ified, no part of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISOs member body in the country of the requester. ISO copyright office C
7、ase postale 56 CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2006 All rights reserved Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Hong Kong Polytechni
8、c Univ/9976803100 Not for Resale, 04/17/2007 09:44:56 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15753:2006(E) ISO 2006 All rights reserved iii Contents Page Foreword iv 1 Scope . 1 2 Normative references. 1 3 Terms and definitions. 1 4 Principle. 2 5 Reagents and
9、materials . 2 6 Apparatus 3 7 Sampling 4 8 Sample preparation 5 9 Procedure for determination of PAHs from fats and oils: General method . 5 9.1 Preliminary remarks . 5 9.2 Blank 5 9.3 Determination of recovery values (without matrix). 5 9.4 Extraction (liquid/liquid extraction) 6 9.5 Purification o
10、n C18-bonded phase cartridge (solid/liquid extraction) 6 9.6 Purification on Florisil-bonded phase cartridge (solid/liquid extraction). 6 10 Procedure for determination of PAHs from fats and oils: Method specific for coconut oil . 7 10.1 First extraction (liquid/liquid extraction) 7 10.2 Second extr
11、action (liquid/liquid extraction). 8 10.3 Purification on C18-bonded phase cartridge (solid/liquid extraction) 8 10.4 Purification on Florisil-bonded phase cartridge (solid/liquid extraction). 8 11 High-performance liquid chromatography (HPLC) . 9 11.1 Operating conditions 9 11.2 Detection parameter
12、s. 10 11.3 Analysis of samples and standards 11 11.4 Confirmation of the presence of PAHs. 12 12 Expression of results . 12 13 Precision 12 13.1 Interlaboratory test. 12 13.2 Repeatability 13 13.3 Reproducibility 13 14 Test report . 13 Annex A (informative) Recovery values, flow charts, chromatogram
13、s and injection sequences. 14 Annex B (informative) Results of the interlaboratory test . 19 Bibliography. 22 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Hong Kong Polytechnic Univ/9976803100 Not for Resale, 04/17/2007 09:44:56 MDTNo repro
14、duction or networking permitted without license from IHS -,-,- ISO 15753:2006(E) iv ISO 2006 All rights reserved Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards
15、 is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take p
16、art in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to
17、 prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility t
18、hat 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. ISO 15753 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal and vegetable fats and oils. Copyright In
19、ternational Organization for Standardization Provided by IHS under license with ISO Licensee=Hong Kong Polytechnic Univ/9976803100 Not for Resale, 04/17/2007 09:44:56 MDTNo reproduction or networking permitted without license from IHS -,-,- INTERNATIONAL STANDARD ISO 15753:2006(E) ISO 2006 All right
20、s reserved 1 Animal and vegetable fats and oils Determination of polycyclic aromatic hydrocarbons 1 Scope This International Standard describes two methods for the determination of 15 polycyclic aromatic hydrocarbons (PAHs) in animal and vegetable fats and oils: a general method, and a method specif
21、ic for coconut oil and vegetable oils with short-chain fatty acids. These methods are not quantitative for the very volatile compounds such as naphthalene, acenaphthene and fluorene. Due to interferences provided by the matrix itself, palm oil and olive pomace oil cannot be analysed using this metho
22、d. 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 0,3 g/kg, and indeno(1,2,3-c,d)pyrene where the quantification limit is 1 g/kg. 2 Normative references The following referenced documents are
23、indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 661, Animal and vegetable fats and oils Preparation of test sample 3 Terms and defin
24、itions For the purposes of this document, the following terms and definitions apply. 3.1 polycyclic aromatic hydrocarbon PAH compound that contains two or more condensed (fused) aromatic hydrocarbon rings and the content of which can be determined according to the method specified in this Internatio
25、nal Standard NOTE 1 The content is given in micrograms per kilogram. NOTE 2 In general PAHs are divided into light PAHs with two to four aromatic rings, and heavy PAHs with five or more aromatic rings. EXAMPLES Light PAHs include: naphthalene (CAS RN 91-20-3), acenaphthene (CAS RN 83-32-9), acenapht
26、hylene (CAS RN 208-96-8), fluorene (CAS RN 86-73-7), anthracene (CAS RN 120-12-7), phenanthrene (CAS RN 85-01-8), fluoranthene (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). Copyright International Organization for Standardization Provide
27、d by IHS under license with ISO Licensee=Hong Kong Polytechnic Univ/9976803100 Not for Resale, 04/17/2007 09:44:56 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15753:2006(E) 2 ISO 2006 All rights reserved Heavy PAHs include: benzo(a)pyrene (CAS RN 50-32-8), benzo(b)f
28、luoranthene (CAS RN 205-99-2), benzo(k)fluoranthene (CAS RN 207-08-9), 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 Principle The polycyclic aromatic hydrocarbons are extracted with an acetonitrile/acetone mixture follow
29、ed by purification on C18 reversed-phase and then Florisil bonded-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 excitat
30、ion and emission wavelengths. 5 Reagents and materials WARNING Attention is drawn to the regulations governing the handling of dangerous matter. Technical, organizational and personal safety measures must be followed. Use only reagents of recognized analytical grade unless otherwise stated. Check th
31、e quality of solvents before use by concentrating the solvent about 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 grade1). 5.2 Hexane, HPLC grade1). 5.3 Acet
32、onitrile, HPLC grade1). 5.4 Acetone, HPLC grade1). 5.5 Dichloromethane, HPLC grade1). 5.6 Toluene, HPLC grade1). 5.7 Water, HPLC grade1). 5.8 Tetrahydrofuran, HPLC grade1). 5.9 Solvent mixture 1: acetonitrile/acetone (volume fraction 60 % / 40 %). Quantity used per sample: 41 ml for general method,
33、36 ml for method specific for coconut oil. 5.10 Solvent mixture 2: acetonitrile/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 f
34、or general method, 2 7 ml for method specific for coconut oil. 1) These can be obtained from, for example, Baker. This information is given for the convenience of users of this International Standard and does not constitute an endorsement by ISO of these products. Equivalent products may be used if
35、they can be shown to lead to the same results. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=Hong Kong Polytechnic Univ/9976803100 Not for Resale, 04/17/2007 09:44:56 MDTNo reproduction or networking permitted without license from IHS -,-,-
36、ISO 15753:2006(E) ISO 2006 All rights reserved 3 5.12 Mixture of tetrahydrofuran/methanol (volume fraction 50 % / 50 %). 5.13 Standard solution with 16 certified EPA Priority PAHs in toluene2), at a concentration of 100 g/ml (100 mg/l): naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthre
37、ne, anthracene, fluoranthene, 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. To be stored at 20 C. Before use, allow the solution to warm up to ambient temperature for at least 1 h
38、. NOTE Acenaphthylene 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
39、 standard solution, 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 cartridges3), 2 g phase, 12 ml capacity. 5.17 Florisil bonded-phase cartridges3), 500 mg phase, 3 ml cap
40、acity. 5.18 Stream of nitrogen, pressure regulated at 34,5 kPa (5 psi, about 1,5 l/min). 6 Apparatus Usual 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, capa
41、ble of attaining at least 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
42、 various excitation and emission wavelengths, and computer-assisted acquisition and data treatment. 6.3 C18 reversed-phase column4), 250 mm in length, 4,6 mm internal diameter, 5 m particles, suitable for PAH analysis. 6.4 Vortex mixer. 6.5 Automatic evaporator5), for 10 ml tube (optional), or water
43、 bath (6.6). 2) This can be obtained from, for example, Promochem. 3) This can be obtained from, for example, Varian. This information is given for the convenience of users of this International Standard and does not constitute an endorsement by ISO of these products. Equivalent products may be used
44、 if they can be shown to lead to the same results. 4) This can be obtained from, for example, Vydac, ref. 201TP54. 5) This can be obtained from, for example, Zymark, Zymark TurboVap LV evaporator. Copyright International Organization for Standardization Provided by IHS under license with ISO License
45、e=Hong Kong Polytechnic Univ/9976803100 Not for Resale, 04/17/2007 09:44:56 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 15753:2006(E) 4 ISO 2006 All rights reserved Recommended operating conditions: temperature of water bath 35 C; nitrogen pressure 34,5 kPa. 6.6 Wat
46、er bath, regulated at 35 C. 6.7 Balance, with readability of 0,1 mg. 6.8 Centrifuge tubes, of 100 ml capacity (one per sample). 6.9 Conical centrifuge tubes, of 11 ml capacity (three per sample), with PTFE septa and closed top screw caps (one per sample). 6.10 Graduated cylinders. 6.11 Microsyringe,
47、 250 l. 6.12 Syringe, 1 000 l. 6.13 Graduated pipette, 5 ml. 6.14 Syringe, 5 ml, equipped with an adapter cap for SPE cartridges. 6.15 Vials for autosampler. 6.16 Microvials, of 250 l capacity, adapted for HPLC system. 6.17 Ultrasonic bath, with water temperature not higher than 40 C. 6.18 Pasteur p
48、ipettes, with cotton wool in the top part to prevent contamination. 6.19 Device composed of stand and pincers6), to hold SPE cartridges or, if available, an automatic SPE work station. NOTE Depending on the SPE sample processing station used, the proposed extraction methods may require slight adaptations (times, pressure, volumes). 6.20 Volumetric flask, of capacity 50 ml. 7 Sampling A representative sample should have been sent to the laboratory. It should not have