EN 17194:2019
(Main)Animal feeding stuffs: Methods of sampling and analysis - Determination of Deoxynivalenol, Aflatoxin B1, Fumonisin B1 & B2, T-2 & HT-2 toxins, Zearalenone and Ochratoxin A in feed materials and compound feed by LC-MS/MS
Animal feeding stuffs: Methods of sampling and analysis - Determination of Deoxynivalenol, Aflatoxin B1, Fumonisin B1 & B2, T-2 & HT-2 toxins, Zearalenone and Ochratoxin A in feed materials and compound feed by LC-MS/MS
This document's method of analysis is applicable for the determination of:
- deoxynivalenol (DON) in the tested range of 100 µg/kg to 3 300 µg/kg,
- aflatoxin B1 (AfB1) in the tested range of 2,5 µg/kg to 440 µg/kg,
- fumonisin B1 (FB1) in the tested range of 690 µg/kg to 7 500 µg/kg,
- fumonisin B2 (FB2) in the tested range of 200 µg/kg to 2 500 µg/kg,
- T-2 toxin in the tested range of 7,5 µg/kg to 360 µg/kg,
- HT-2 toxin in the tested range of 14 µg/kg to 1 800 µg/kg,
- zearalenone (ZEN) in the tested range of 30 µg/kg to 600 µg/kg, and
- ochratoxin A (OTA) in the tested range of 10 µg/kg to 230 µg/kg
in cereals and cereal-based compound feed by liquid-chromatography tandem mass spectrometry (LC-MS/MS). The actual working ranges could extend beyond the tested ranges.
Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von Deoxynivalenol, Aflatoxin B1, Fumonisin B1 und B2, T-2- und HT-2-Toxine, Zearalenon und Ochratoxin A in Einzelfuttermitteln und Mischfuttermitteln mittels LC-MS/MS
Das Analyseverfahren dieses Dokumentes ist gültig für die Bestimmung von
— Deoxynivalenol (DON) im untersuchten Bereich von 100 µg/kg bis 3 300 µg/kg,
— Aflatoxin B1 (AfB1) im untersuchten Bereich von 2,5 µg/kg bis 440 µg/kg,
— Fumonisin B1 (FB1) im untersuchten Bereich von 690 µg/kg bis 7 500 µg/kg,
— Fumonisin B2 (FB2) im untersuchten Bereich von 200 µg/kg bis 2 500 µg/kg,
— T 2 Toxin im untersuchten Bereich von 7,5 µg/kg bis 360 µg/kg,
— HT 2 Toxin im untersuchten Bereich von 14 µg/kg bis 1 800 µg/kg,
— Zearalenon (ZEN) im unter¬suchten Bereich von 30 µg/kg bis 600 µg/kg und
— Ochratoxin A (OTA) im untersuchten Bereich von 10 µg/kg bis 230 µg/kg
in Getreide und Getreidemisch¬futtermittel durch Flüssigchromatographie mit Tandem Massen-spektrometrie (LC MS/MS). Die tatsächlichen Arbeitsbereiche könnten über die geprüften Bereiche hinausgehen.
Aliments des animaux : Méthodes d’échantillonnage et d’analyse — Détermination du déoxynivalénol, de l’aflatoxine B1, de la fumonisine B1 et B2, des toxines T-2 et HT-2, de la zéaralénone et de l’ochratoxine A dans les matières premières pour aliments et les aliments composés pour animaux par CL-SM/SM
La méthode d’analyse décrite dans le présent document s’applique à la détermination :
— du déoxynivalénol (DON) dans la plage soumise à essai allant de 100 µg/kg à 3 300 µg/kg ;
— de l’aflatoxine B1 (AfB1) dans la plage soumise à essai allant de 2,5 µg/kg à 440 µg/kg ;
— de la fumonisine B1 (FB1) dans la plage soumise à essai allant de 690 µg/kg à 7 500 µg/kg ;
— de fumonisine B21 (FB2) dans la plage soumise à essai allant de 200 µg/kg à 2 500 µg/kg ;
— de la toxine T-2 dans la plage soumise à essai allant de 7,5 µg/kg à 360 µg/kg ;
— de la toxine HT-2 dans la plage soumise à essai allant de 14 µg/kg à 1 800 µg/kg ;
— de la zéaralénone (ZEN) dans la plage soumise à essai allant de 30 µg/kg à 600 µg/kg ; et
— de l’ochratoxine A (OTA) dans la plage soumise à essai allant de 10 µg/kg à 230 µg/kg.
dans les céréales et les aliments composés pour animaux à base de céréales, par chromatographie en phase liquide couplée à de la spectrométrie de masse (CL-SM/SM). Il se peut que les plages de travail en routine s’étendent au-delà des plages soumises à essai.
Krma: metode vzorčenja in analize - Določevanje deoksinivalenola, aflatoksina B1, fumonizina B1 in B2, toksinov T-2 in HT-2, zearalenona in ohratoksina A v sestavinah krme in krmni mešanici z LC-MS/MS
Ta analitska metoda evropskega standarda se uporablja za določanje deoksinivalenola (DON) v preskusnem obsegu od 96,2 µg/kg do 3269 µg/kg, aflatoksina B1 (AfB1) v preskusnem obsegu od 2,62 µg/kg do 444 µg/kg, fumonisina B1 (FB1) v preskusnem obsegu od 693 µg/kg do 7529 µg/kg, fumonisina B2 (FB2) v preskusnem obsegu od 203 µg/kg do 2465 µg/kg, T- 2 toksina v preskusnem obsegu od 7,47 µg/kg do 360 µg/kg in HT-2 toksina v preskusnem obsegu od 13,9 µg/kg do 1758 µg/kg, zeralenona (ZON) v preskusnem obsegu od 34,3 µg/kg do 593 µg/kg ter ohratoksina A (OTA) v preskusnem obsegu od 10,8 µg/kg do 228 µg/kg pri žitih in krmnih mešanicah na osnovi žit s tandemsko masno spektrometrijo s tekočinsko kromatografijo (LC-MS/MS). Dejanski delovni obsegi lahko presegajo presušene obsege.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-januar-2020
Krma: metode vzorčenja in analize - Določevanje deoksinivalenola, aflatoksina B1,
fumonizina B1 in B2, toksinov T-2 in HT-2, zearalenona in ohratoksina A v
sestavinah krme in krmni mešanici z LC-MS/MS
Animal feeding stuffs: Methods of sampling and analysis - Determination of
Deoxynivalenol, Aflatoxin B1, Fumonisin B1 & B2, T-2 & HT-2 toxins, Zearalenone and
Ochratoxin A in feed materials and compound feed by LC-MS/MS
Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von
Deoxynivalenol, Aflatoxin B1, Fumonisin B1 und B2, T-2- und HT-2-Toxine, Zearalenon
und Ochratoxin A in Einzelfuttermitteln und Mischfuttermitteln mittels LC-MS/MS
Aliments des animaux : Méthodes d’échantillonnage et d’analyse — Détermination du
déoxynivalénol, de l’aflatoxine B1, de la fumonisine B1 et B2, des toxines T-2 et HT-2, de
la zéaralénone et de l’ochratoxine A dans les matières premières pour aliments et les
aliments composés pour animaux par CL-SM/SM
Ta slovenski standard je istoveten z: EN 17194:2019
ICS:
65.120 Krmila Animal feeding stuffs
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 17194
EUROPEAN STANDARD
NORME EUROPÉENNE
November 2019
EUROPÄISCHE NORM
ICS 65.120
English Version
Animal feeding stuffs: Methods of sampling and analysis -
Determination of Deoxynivalenol, Aflatoxin B1, Fumonisin
B1 & B2, T-2 & HT-2 toxins, Zearalenone and Ochratoxin A
in feed materials and compound feed by LC-MS/MS
Aliments des animaux : Méthodes d'échantillonnage et Futtermittel: Probenahme- und
d'analyse - Détermination du déoxynivalénol, de Untersuchungsverfahren - Bestimmung von
l'aflatoxine B1, de la fumonisine B1 et B2, des toxines Deoxynivalenol, Aflatoxin B1, Fumonisin B1 und B2, T-
T-2 et HT-2, de la zéaralénone et de l'ochratoxine A 2- und HT-2-Toxine, Zearalenon und Ochratoxin A in
dans les matières premières pour aliments et les Einzelfuttermitteln und Mischfuttermitteln mittels LC-
aliments composés pour animaux par CL-SM/SM MS/MS
This European Standard was approved by CEN on 9 September 2019.
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-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 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,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17194:2019 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
1 Scope . 4
2 Normative references . 4
3 Terms and definitions . 4
4 Principle . 4
5 Reagents . 5
6 Apparatus . 7
7 Procedure. 8
7.1 Sample preparation . 8
7.2 Extraction . 9
7.3 Test solution . 9
7.4 Spiking procedure . 10
8 Measurements . 10
8.1 General . 10
8.2 LC conditions. 10
8.3 MS conditions . 10
8.4 Batch composition . 11
8.5 Peak identification . 11
8.6 Determination of mycotoxins in calibration and test solutions . 11
8.7 Calibration . 11
9 Determination of mass fraction . 12
10 Precision . 13
10.1 Interlaboratory study . 13
10.2 Repeatability . 13
10.3 Reproducibility . 14
11 Test report . 14
Annex A (informative) Precision data . 15
Annex B (informative) Examples . 25
Annex C (informative) Example chromatograms . 30
Bibliography . 32
European foreword
This document (EN 17194:2019) has been prepared by 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 May 2020, and conflicting national standards shall be
withdrawn at the latest by May 2020.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document has been prepared under a standardization request given to CEN by the European
Commission and the European Free Trade Association.
According to the CEN-CENELEC Internal Regulations, the national standards organisations 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, Republic of North
Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United
Kingdom.
1 Scope
This document's method of analysis is applicable for the determination of:
— deoxynivalenol (DON) in the tested range of 100 µg/kg to 3 300 µg/kg,
— aflatoxin B1 (AfB1) in the tested range of 2,5 µg/kg to 440 µg/kg,
— fumonisin B1 (FB1) in the tested range of 690 µg/kg to 7 500 µg/kg,
— fumonisin B2 (FB2) in the tested range of 200 µg/kg to 2 500 µg/kg,
— T-2 toxin in the tested range of 7,5 µg/kg to 360 µg/kg,
— HT-2 toxin in the tested range of 14 µg/kg to 1 800 µg/kg,
— zearalenone (ZEN) in the tested range of 30 µg/kg to 600 µg/kg, and
— ochratoxin A (OTA) in the tested range of 10 µg/kg to 230 µg/kg
in cereals and cereal-based compound feed by liquid-chromatography tandem mass spectrometry (LC-
MS/MS). The actual working ranges could extend beyond the tested ranges.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements 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.
EN ISO 3696:1995, Water for analytical laboratory use — Specification and test methods (ISO 3696:1987)
EN ISO 6498, Animal feeding stuffs — Guidelines for sample preparation (ISO 6498)
3 Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— IEC Electropedia: available at http://www.electropedia.org/
— ISO Online browsing platform: available at https://www.iso.org/obp.
4 Principle
A test portion of finely ground and homogeneous material is extracted by shaking with a mixture of
acetonitrile and aqueous formic acid solution. The extract is centrifuged and an aliquot of the supernatant
extract is transferred to a deactivated glass vial, mixed with an appropriate amount of stable-isotope
labelled analogues and evaporated to dryness. The reconstituted sample is filtered and quantified with a
Liquid Chromatography - Mass Spectrometry (LC-MS) system. Laboratories using this method shall
demonstrate the following limits of quantitation (LOQs) in order to be able to apply this method over the
whole validation range: for DON ≤ 100 µg/kg, for AfB1 ≤ 2 µg/kg, for FB1 and FB2 ≤ 500 µg/kg
(FB1 ≤ 375 µg/kg and FB2 ≤ 125 µg/kg), for T-2 and HT-2 toxin ≤ 10 µg/kg, for ZEN ≤ 20 µg/kg and for
OTA ≤ 10 µg/kg.
NOTE The performance data collected in this study came from laboratories that verified to be able to achieve
the above mentioned LOQs. Applicants that could not verify this, were excluded in order to ensure that validation
data could be obtained over the targeted concentration range.
5 Reagents
Use only reagents of recognized analytical grade and water complying with grade 1 of EN ISO 3696:1995,
unless otherwise specified. Commercially available solutions with equivalent properties to those listed
may be used.
WARNING — Environmental regulations and rules apply when disposing of waste solvents.
Decontamination procedures for laboratory wastes have been reported by the International Agency for
Research on Cancer (IARC), see [1, 2].
5.1 Water (deionized).
5.2 Water, LC-MS grade or of comparable purity (e.g. resistance of 18,2 MΩcm or respectively
conductivity of 55 nS/cm at 20 °C).
5.3 Methanol, LC-MS grade.
5.4 Formic acid (FA), 98-100 %.
5.5 Acetonitrile (ACN), LC grade.
5.6 Extraction solvent composed of 20 parts water (5.1), 79 parts acetonitrile (5.5) and 1 part formic
acid (5.4) (v/v/v).
5.7 20 % acetic acid solution, for washing glassware.
5.8 Aflatoxin B1 (AfB1), purity ≥ 95 %.
5.9 Deoxynivalenol (DON), purity ≥ 95 %.
5.10 Fumonisin B1 (FB1), purity ≥ 95 %.
5.11 Fumonisin B2 (FB2), purity ≥ 95 %.
5.12 HT-2 toxin (HT-2), purity ≥ 95 %.
5.13 T-2 toxin (T-2), purity ≥ 95 %.
5.14 Zearalenone (ZEN), purity ≥ 95 %.
5.15 Ochratoxin A (OTA), purity ≥ 95 %.
13 13
5.16 C -Aflatoxin B1 ( C -AfB1).
17 17
13 13
5.17 C -Deoxynivalenol ( C -DON).
15 15
13 13
5.18 C -Fumonisin B1 ( C -FB1).
34 34
13 13
5.19 C -Fumonisin B2 ( C -FB2).
34 34
13 13
5.20 C -HT-2 toxin ( C -HT2).
22 22
13 13
5.21 C -T-2 toxin ( C -T2).
24 24
13 13
5.22 C -Zearalenone ( C -ZEN).
18 18
13 13
5.23 C -Ochratoxin A ( C -OTA).
20 20
5.24 Stock standard solution
A mixture containing Deoxynivalenol (5.9), Aflatoxin B1 (5.8), Fumonisin B1 (5.10) and Fumonisin B2
(5
...
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