EN ISO 15302:2010
(Main)Animal and vegetable fats and oils - Determination of benzo[a]pyrene - Reverse-phase high performance liquid chromatography method (ISO 15302:2007)
Animal and vegetable fats and oils - Determination of benzo[a]pyrene - Reverse-phase high performance liquid chromatography method (ISO 15302:2007)
ISO 15302:2007 specifies a method for the determination of benzo[a]pyrene in crude or refined edible oils and fats by reverse-phase high performance liquid chromatography (HPLC) using fluorimetric detection in the range 0,1 µg/kg to 50 µg/kg.
Tierische und pflanzliche Fette und Öle - Bestimmung des Benzo[a]pyren-Gehalts - Umkehrphasen-Hochleistungsflüssigchromatographisches Verfahren (ISO 15302:2007)
Diese Internationale Norm legt ein Verfahren für die Bestimmung von Benzo[a]pyren in rohen oder raffinierten
essbaren Ölen und Fetten durch Hochleistungsflüssigchromatographie mit umgekehrten Phasen (HPLC)
unter Anwendung eines fluorimetrischen Nachweises in dem Bereich von 0,1 µg/kg bis 50 µg/kg fest.
Corps gras d'origines animale et végétale - Détermination du benzo[a]pyrène - Méthode par chromatographie liquide à haute performance à polarité de phase inversée (ISO 15302:2007)
L'ISO 15302:2007 spécifie une méthode de détermination du benzo[a]pyrène dans les corps gras comestibles bruts ou raffinés par chromatographie liquide à haute performance (CLHP) à polarité de phase inversée à l'aide d'une détection fluorimétrique, dans la gamme de 0,1 µg/kg à 50 µg/kg.
Živalske in rastlinske maščobe in olja - Določevanje benzo[a]pirena - Metoda tekočinske kromatografije visoke ločljivosti z reverzno fazo (ISO 15302:2007)
Ta mednarodni standard določa metodo za določevanje benzo[a]pirena v surovih ali rafiniranih jedilnih oljih in maščobah z tekočinsko kromatografijo visoke ločljivosti z reverzno fazo (HPLC), ki uporablja fluorimetrično detekcijo v razponu 0,1 µg/kg do 50 µg/kg.
General Information
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Standards Content (Sample)
SLOVENSKI STANDARD
01-september-2010
1DGRPHãþD
SIST EN ISO 15302:2007
äLYDOVNHLQUDVWOLQVNHPDãþREHLQROMD'RORþHYDQMHEHQ]R>D@SLUHQD0HWRGD
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Animal and vegetable fats and oils - Determination of benzo[a]pyrene - Reverse-phase
high performance liquid chromatography method (ISO 15302:2007)
Tierische und pflanzliche Fette und Öle - Bestimmung des Benzo[a]pyren-Gehalts -
Umkehrphasen-Hochleistungsflüssigchromatographisches Verfahren (ISO 15302:2007)
Corps gras d'origines animale et végétale - Détermination du benzo[a]pyrène - Méthode
par chromatographie liquide à haute performance à polarité de phase inversée (ISO
15302:2007)
Ta slovenski standard je istoveten z: EN ISO 15302:2010
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EUROPEAN STANDARD
EN ISO 15302
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2010
ICS 67.200.10 Supersedes EN ISO 15302:2007
English Version
Animal and vegetable fats and oils - Determination of
benzo[a]pyrene - Reverse-phase high performance liquid
chromatography method (ISO 15302:2007)
Corps gras d'origines animale et végétale - Détermination Tierische und pflanzliche Fette und Öle - Bestimmung des
du benzo[a]pyrène - Méthode par chromatographie liquide Benzo[a]pyren-Gehalts - Umkehrphasen-
à haute performance à polarité de phase inversée (ISO Hochleistungsflüssigchromatographisches Verfahren (ISO
15302:2007) 15302:2007)
This European Standard was approved by CEN on 18 March 2010.
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 alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN 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 by translation
under the responsibility of a CEN member into its own language and notified to the CEN Management Centre has the same status as the
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CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
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Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2010 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 15302:2010: E
worldwide for CEN national Members.
Contents Page
Foreword .3
Foreword
The text of ISO 15302:2007 has been prepared by Technical Committee ISO/TC 34 “Food products” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 15302:2010 by
Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products -
Methods of sampling and analysis” the secretariat 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 2010, and conflicting national standards shall be withdrawn at
the latest by October 2010.
Attention is 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 15302:2007.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia,
Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain,
Sweden, Switzerland and the United Kingdom.
Endorsement notice
The text of ISO 15302:2007 has been approved by CEN as a EN ISO 15302:2010 without any modification.
INTERNATIONAL ISO
STANDARD 15302
Second edition
2007-12-15
Animal and vegetable fats and oils —
Determination of benzo[a]pyrene —
Reverse-phase high performance liquid
chromatography method
Corps gras d'origines animale et végétale — Détermination du
benzo[a]pyrène — Méthode par chromatographie liquide à haute
performance à polarité de phase inversée
Reference number
ISO 15302:2007(E)
©
ISO 2007
ISO 15302:2007(E)
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ii © ISO 2007 – All rights reserved
ISO 15302:2007(E)
Foreword
ISO (the International 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 has 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 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 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 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.
ISO 15302 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
This second edition cancels and replaces the first edition (ISO 15302:1998).
INTERNATIONAL STANDARD ISO 15302:2007(E)
Animal and vegetable fats and oils — Determination of
benzo[a]pyrene — Reverse-phase high performance liquid
chromatography method
1 Scope
This International Standard specifies a method for the determination of benzo[a]pyrene in crude or refined
edible oils and fats by reverse-phase high performance liquid chromatography (HPLC) using fluorimetric
detection in the range 0,1 µg/kg to 50 µg/kg.
2 Normative references
The following referenced documents are 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 definitions
For the purposes of this document, the following terms and definitions apply.
3.1
benzo[a]pyrene content
mass fraction of benzo[a]pyrene in the test portion, as determined using the method specified in this
International Standard
NOTE The content is expressed in micrograms per kilogram.
4 Principle
A test portion is dissolved in light petroleum and benzo[b]chrysene is added as internal standard. A suitable
amount of sample is adsorbed on an alumina column, and eluted with light petroleum to remove any
benzo[a]pyrene present.
5 Reagents
Use only reagents of recognized analytical grade, unless otherwise specified. Where analytical grade solvents
other than the recommended ones are used, a full blank analysis shall be carried out and the results of this
blank analysis reported.
SAFETY PRECAUTIONS — Attention is drawn to regulations which specify handling procedures for
dangerous substances. Users should be aware of and comply with technical, organizational, and
personal safety measures.
ISO 15302:2007(E)
5.1 Water, double distilled, filtered through a membrane filter of pore size 0,45 µm; deionized water
obtained by purifying demineralized water systems may also be used.
5.2 Light petroleum (boiling point range 40 °C to 60 °C), or hexane, redistilled over potassium hydroxide
pellets (4 g/l).
5.3 Acetonitrile, suitable for HPLC.
5.4 Tetrahydrofuran, suitable for HPLC.
5.5 Acetonitrile-tetrahydrofuran mixture, prepared by mixing 90 ml acetonitrile (5.3) and 10 ml
tetrahydrofuran (5.4).
5.6 Toluene, suitable for HPLC.
5.7 Sodium sulfate, granular, anhydrous.
1)
5.8 Alumina activity grade 4, prepared from neutral aluminium oxide, activity grade super 1 , deactivated
by the addition of 10 ml water (5.1) to 90 g of alumina.
Due to the differences in activity of alumina of various brands, a check is recommended to confirm that the
deactivation procedure is appropriate for total benzo[a]pyrene recovery from a reference sample.
CAUTION — THE DEACTIVATION REACTION IS EXOTHERMIC AND PRESSURE MAY BUILD UP.
Shake the container for about 15 min and allow the contents to equilibrate for 24 h. Store the alumina in a
closed vessel at ambient temperature.
2)
5.9 Benzo[a]pyrene , of purity 99,0 % by mass.
CAUTION — BENZO[a]PYRENE IS A KNOWN CARCINOGEN. CARRY OUT ALL WORK WITH IT IN A
FUME HOOD, WEARING GLOVES TO MINIMIZE EXPOSURE.
5.9.1 Benzo[a]pyrene stock solution in toluene, 0,5 mg/ml.
Weigh, to the nearest 0,1 mg, about 12,5 mg of benzo[a]pyrene in a 25 ml graduated flask. Dissolve it in
toluene (5.6) and make up to the mark with that solvent.
Store the solution in the dark at 4 °C where it is stable for at least 6 months.
5.9.2 Benzo[a]pyrene standard solutions.
Prepare two standard solutions containing approximately 0,2 µg/ml and 0,01 µg/ml of benzo[a]pyrene,
respectively by diluting aliquots of the stock solution (5.9.1) with acetonitrile.
1) “Aluminium oxide 90 active neutral” is an example of a suitable product available commercially. This information is
given for the convenience of users of this International Standard and does not constitute an endorsement by ISO of this
product. Equivalent products may be used if they can be shown to lead to the same results.
2) A suitable reference material is available from the Joint Research Centre of the European Commission, Institute for
Reference Materials and Measurements (IRMM). This information is given for the convenience of users of this
International Standard and does not constitute an endorsement by ISO of this product.
2 © ISO 2007 – All rights reserved
ISO 15302:2007(E)
3)
5.10 Benzo[b]chrysene internal standard solution in acetonitrile.
Prepare a stock solution containing, to the nearest nanogram, approximately 10 ng/µl. Dilute this solution by a
factor of 10 in a volumetric flask, to obtain an internal standard solution with a concentration of approximately
1 ng/µl.
3)
NOTE This solution may also be prepared by dissolving benzo[b]chrysene , 99,0 % by mass, in acetonitrile.
6 Apparatus
Usual laboratory apparatus and, in particular, the following.
6.1 Glass column for chromatography, of length 300 mm and internal diameter 15 mm, fitted with
sintered glass discs, and polytetrafluoroethylene (PTFE) tap.
6.2 Water baths, one maintained at (35 ± 2) °C and another at (65 ± 2) °C.
6.3 Flash evaporator, a rotary evaporator with vacuum and a water bath set at 40 °C may be used. Care
should be taken to prevent cross contamination. Clean the system thoroughly between determinations.
6.4 High performance liquid chromatograph, consisting of an HPLC pump, injection valve with 50 µl
sample loop, reverse-phase column, electronic integrator and chart recorder.
If an autosampler is used, the sample loop shall be flushed with aceton
...
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