EN ISO 17375:2006
(Main)Animal feeding stuffs - Determination of aflatoxin B1 (ISO 17375:2006)
Animal feeding stuffs - Determination of aflatoxin B1 (ISO 17375:2006)
ISO 17375:2006 specifies a method for the determination of aflatoxin B1 in animal feeding stuffs using high-performance liquid chromatography with post-column derivatization.
It is applicable to animal feeding stuffs with a fat content of up to 50 %.
The limit of quantification of this method has been demonstrated to be better than 0,5 g/kg for aflatoxin B1 for a signal-to-noise ratio of 6.
Futtermittel - Bestimmung von Aflatoxin B1 (ISO 17375:2006)
Diese Internationale Norm legt ein Verfahren zur Bestimmung von Aflatoxin B1 in Futtermitteln mittels Hochleistungs-Flüssigchromatographie mit Nachsäulenderivatisierung fest.
Das Verfahren ist anwendbar für Futtermittel mit einem Fettgehalt bis zu 50 %.
Die quantitative Nachweisgrenze dieses Verfahrens wurde bei einem Signal-Rausch-Verhältnis von 6 für Aflatoxin B1 als besser als 0,5 µg/kg nachgewiesen.
ANMERKUNG Das Verfahren basiert auf dem in der Literaturstelle [1] angegebenen Verfahren.
Aliments des animaux - Dosage de l'aflatoxine B1 (ISO 17375:2006)
L'ISO 17375:2006 spécifie une méthode de dosage de l'aflatoxine B1 dans les aliments des animaux en utilisant la chromatographie liquide haute performance avec dérivation post-colonne.
Elle est applicable aux aliments pour animaux dont la teneur en graisse est inférieure ou égale à 50 %.
Il a été démontré que la limite de quantification de l'aflatoxine B1 par cette méthode est inférieure à 0,5 g/kg avec un rapport signal/bruit de 6.
Krma – Določevanje aflatoksina B1 (ISO 17375:2006)
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-september-2006
.UPD±'RORþHYDQMHDIODWRNVLQD% ,62
Animal feeding stuffs - Determination of aflatoxin B1 (ISO 17375:2006)
Futtermittel - Bestimmung von Aflatoxin B1 (ISO 17375:2006)
Aliments des animaux - Dosage de l'aflatoxine B1 (ISO 17375:2006)
Ta slovenski standard je istoveten z: EN ISO 17375:2006
ICS:
65.120 Krmila Animal feeding stuffs
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EUROPEAN STANDARD
EN ISO 17375
NORME EUROPÉENNE
EUROPÄISCHE NORM
June 2006
ICS 65.120
English Version
Animal feeding stuffs - Determination of aflatoxin B1 (ISO
17375:2006)
Aliments des animaux - Dosage de l'aflatoxine B1 (ISO Futtermittel - Bestimmung von Aflatoxin B1 (ISO
17375:2006) 17375:2006)
This European Standard was approved by CEN on 12 May 2006.
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 Central Secretariat 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 Central Secretariat has the same status as the official
versions.
CEN members are the national standards bodies of Austria, Belgium, Cyprus, Czech Republic, Denmark, Estonia, Finland, France,
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Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
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© 2006 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 17375:2006: E
worldwide for CEN national Members.
Foreword
This document (EN ISO 17375:2006) has been prepared by Technical Committee ISO/TC 34
"Agricultural food products" in collaboration with Technical Committee CEN/TC 327 "Animal
feeding stuffs - Methods of sampling and analysis", the secretariat of which is held by NEN.
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 December 2006, and conflicting national
standards shall be withdrawn at the latest by December 2006.
According to the CEN/CENELEC Internal Regulations, the national standards organizations of
the following countries are bound to implement this European Standard: Austria, Belgium,
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 United Kingdom.
Endorsement notice
The text of ISO 17375:2006 has been approved by CEN as EN ISO 17375:2006 without any
modifications.
INTERNATIONAL ISO
STANDARD 17375
First edition
2006-06-01
Animal feeding stuffs — Determination
of aflatoxin B
Aliments des animaux — Dosage de l'aflatoxine B
Reference number
ISO 17375:2006(E)
©
ISO 2006
ISO 17375:2006(E)
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ii © ISO 2006 – All rights reserved
ISO 17375:2006(E)
Contents Page
Foreword. iv
1 Scope. 1
2 Normative references. 1
3 Principle. 1
4 Reagents. 1
5 Apparatus. 4
6 Procedures. 6
6.1 Conditioning of IACs . 6
6.2 Extraction. 6
6.3 Immunoaffinity clean up . 6
6.4 Post-column derivatization. 7
6.5 Calibration curve. 7
6.6 Calculation. 7
6.7 Spiking procedures for recovery determination .8
7 Precision. 8
7.1 Interlaboratory test . 8
7.2 Repeatability. 8
7.3 Reproducibility. 9
8 Test report. 9
Annex A (informative) Results of an interlaboratory test. 10
Bibliography . 11
ISO 17375:2006(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 17375 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 10, Animal
feeding stuffs.
iv © ISO 2006 – All rights reserved
INTERNATIONAL STANDARD ISO 17375:2006(E)
Animal feeding stuffs — Determination of aflatoxin B
1 Scope
This International Standard specifies a method for the determination of aflatoxin B in animal feeding stuffs
using high-performance liquid chromatography with post-column derivatization.
It is applicable to animal feeding stuffs with a fat content of up to 50 %.
The limit of quantification of this method has been demonstrated to be better than 0,5 µg/kg for aflatoxin B for
a signal-to-noise ratio of 6.
NOTE The method is based on that given in Reference [1].
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 3696:1987, Water for analytical laboratory use — Specification and test methods
3 Principle
A test portion is extracted with a solvent solution (acetone/water). The sample extract is filtered, diluted with
water or phosphate-buffered saline to a specified solvent concentration. A test portion is applied on an
immunoaffinity column (IAC) containing antibodies specific to aflatoxin B . The aflatoxin B is removed from
1 1
the IAC with neat methanol, and then quantified by reverse-phase high-performance liquid chromatography
(RP-HPLC) with post-column derivatization (PCD) involving bromination. The PCD is achieved with either
electrochemically generated bromine or with pyridinium hydrobromide perbromide (PBPB) followed by
fluorescence detection.
4 Reagents
Use only reagents of recognized analytical grade, unless otherwise specified.
WARNING — This method requires the use of toxic inflammable liquids such as acetone, methanol
and acetonitrile. Avoid contact and keep away from heat, sparks or open flames.
[2],[3]
NOTE Decontamination procedures for laboratory wastes have been developed and validated by the
International Agency for Research on Cancer (WHO).
4.1 Water, complying with grade 3 in accordance with ISO 3696:1987.
4.2 Phosphate buffer saline (PBS), pH 7,4.
ISO 17375:2006(E)
PBS may be prepared from potassium chloride (0,20 g), potassium dihydrogen phosphate (0,20 g), anhydrous
disodium hydrogen phosphate (1,16 g) [or disodium hydrogen phosphate dodecahydrate (2,92 g)] and sodium
chloride (8,00 g) added to 900 ml purified water. Adjust the pH to pH 7,4 (with 0,1 mol/l HCl or 0,1 mol/l NaOH
as appropriate) and dilute the solution to 1,0 l.
Alternatively, commercially available phosphate-buffered saline tablets with equivalent properties may be used.
PBS is not microbiologically stable and should be prepared fresh at least once a week.
4.3 Pyridinium hydrobromide perbromide (PBPB, CAS: 39416-48-3).
This reagent is not required in the case of using electrochemically generated bromine.
4.4 Potassium bromide.
This reagent is not required in the case of using the PBPB reagent.
4.5 HPLC-grade acetonitrile.
4.6 HPLC-grade methanol.
4.7 Acetone, pure.
4.8 HPLC grade water, complying with grade 1 of ISO 3696:1987.
4.9 Extraction solvent, solution of acetone (4.7) and water (4.8) [85+15 (by volume)].
4.10 Nitric acid, c(HNO ) = 4 mol/l.
This reagent is not required in the case of using the PBPB reagent.
4.11 Immunoaffinity column (IAC)
The IAC should contain antibodies raised against aflatoxin B . The IAC should have a capacity of not less
than 40 ng of aflatoxin B and should give a recovery of not less than 80 % for aflatoxin B when applied as a
standard solution in acetone/water containing 0,25 ng of aflatoxin B .
4.12 HPLC mobile phase solvent A, for use with PBPB post column reagent only.
Use a solution of water (4.8)/acetonitrile (4.5)/methanol (4.6) [6+2+3 (by volume)]. The ratio of solvents may
be adjusted to give optimum separation parameters.
4.13 HPLC mobile phase solvent B, for use with electrochemically generated bromine only.
Use a solution of water (4.8)/acetonitrile (4.5)/methanol (4.6) [6+2+3 (by volume)] containing 120 mg
potassium bromide (4.4) and 350 µl nitric acid at 4 mol/l (4.10) per litre of mobile phase. The ratio of solvents
may be adjusted to give optimum separation parameters.
The mobile phase solvents (4.12 and 4.13) should be degassed.
4.14 Post-column reagent, for use with PBPB post colum
...
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