EN 17425:2021
(Main)Foodstuffs - Determination of ergot alkaloids in cereals and cereal products by dSPE clean-up and HPLC-MS/MS
Foodstuffs - Determination of ergot alkaloids in cereals and cereal products by dSPE clean-up and HPLC-MS/MS
This document describes a method for the determination of the sum of six ergot alkaloids (ergocornine, ergometrine, ergocristine, ergotamine, ergosine and ergocryptine) and their -inine epimer pairs by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) after clean-up by dispersive solid phase extraction (dSPE).
The method has been validated in the range 13,2 µg/kg to 168 µg/kg for the sum of the twelve ergot alkaloids, in rye flour, rye bread and cereal products (breakfast cereal, infant breakfast cereal, and crispbread) that contained rye as an ingredient, as well as seeded wholemeal flour and a barley and rye flour mixture.
Method performance was satisfactory in the range 24,1 µg/kg to 168 µg/kg, however at lower concentrations RSDR values were greater than 44 %, and HorRat values exceeded 2,0, indicating the method may not be fully suitable at concentrations below 24 µg/kg for sum of ergot alkaloids, although it is suitable for screening at these concentrations.
Lebensmittel - Bestimmung von Ergotalkaloiden in Getreiden und Getreideerzeugnissen mit dSPE-Reinigung und LC-MS/MS
Dieses Dokument beschreibt ein Verfahren für die Bestimmung der Summe von sechs Ergotalkaloiden (Ergocornin, Ergometrin, Ergocristin, Ergotamin, Ergosin und Ergocryptin) und ihrer -inin-Epimerenpaare, wobei die Flüssigchromatographie mit Tandem-Massenspektrometrie (LC-MS/MS) nach Reinigung durch dispersive Festphasenextraktion (dSPE) zum Einsatz kommt.
Dieses Verfahren wurde für die Summe von zwölf Ergotalkaloiden im Bereich von 13,2 μg/kg bis 168 μg/kg in Roggenmehl, Roggenbrot und Getreideerzeugnissen (Frühstückscerealien, Frühstückscerealien für Kleinkinder und Knäckebrot), die Roggen als Inhaltsstoff enthielten, wie auch in Vollkornmehl und einer Mehlmischung aus Roggen und Gerste validiert.
Das Verfahren erbrachte zufriedenstellende Ergebnisse im Bereich von 24,1 μg/kg bis 168 μg/kg, allerdings betrugen die RSDR-Werte bei den niedrigeren Konzentrationen mehr als 44 % und die HorRat-Werte überschritten 2,0, was darauf hinweist, dass das Verfahren möglicherweise als nicht zufriedenstellend bei Konzentrationen unter 24 μg/kg für die Summe der Ergotalkaloide zu bewerten ist. Dennoch ist das Verfahren bei diesen Konzentrationen zum Screening geeignet.
Produits alimentaires - Dosage des alcaloïdes de l’ergot dans les céréales et les produits céréaliers par purification par dSPE et CL-SM/SM
Le présent document décrit une méthode pour le dosage de la somme des six alcaloïdes de l’ergot (ergocornine, ergométrine, ergocristine, ergotamine, ergosine et ergocryptine) et de leurs épimères ( inines) par chromatographie liquide couplée à la spectrométrie de masse en tandem (CL-SM/SM) après purification par extraction sur phase solide dispersée (dSPE).
La méthode a été validée dans la gamme de 13,2 µg/kg à 168 µg/kg pour la somme des douze alcaloïdes de l’ergot, dans la farine de seigle, le pain de seigle et les produits céréaliers (céréales pour petit-déjeuner, céréales pour nourrissons et biscottes) contenant du seigle, ainsi que dans la farine complète aux graines et un mélange de farines de seigle et d’orge.
La performance de la méthode s’est révélée satisfaisante dans la gamme de 24,1 µg/kg à 168 µg/kg, cependant à des concentrations inférieures, les valeurs de RSDR étaient supérieures à 44 % et les valeurs de HorRat dépassaient 2,0, ce qui indique que la méthode peut ne pas être parfaitement adaptée à des concentrations de moins de 24 µg/kg pour la somme des alcaloïdes de l’ergot, bien qu’elle soit adaptée au criblage à ces concentrations.
Živila - Določevanje alkaloidov rženih rožičkov (ergot) v žitu in žitnih proizvodih s čiščenjem dSPE in LC-MS/MS
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-julij-2021
Živila - Določevanje alkaloidov rženih rožičkov (ergot) v žitu in žitnih proizvodih s
čiščenjem dSPE in LC-MS/MS
Foodstuffs - Determination of ergot alkaloids in cereals and cereal products by dSPE
clean-up and LC-MS/MS
Lebensmittel - Bestimmung von Ergotalkaloiden in Getreiden und Getreideerzeugnissen
mit dSPE-Reinigung und LC-MS/MS
Produits alimentaires - Dosage des alcaloïdes de l’ergot dans les céréales et les produits
céréaliers par purification par dSPE et CL-SM/SM
Ta slovenski standard je istoveten z: EN 17425:2021
ICS:
67.050 Splošne preskusne in General methods of tests and
analizne metode za živilske analysis for food products
proizvode
67.060 Žita, stročnice in proizvodi iz Cereals, pulses and derived
njih products
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 17425
EUROPEAN STANDARD
NORME EUROPÉENNE
April 2021
EUROPÄISCHE NORM
ICS 67.050; 67.060
English Version
Foodstuffs - Determination of ergot alkaloids in cereals
and cereal products by dSPE clean-up and HPLC-MS/MS
Produits alimentaires - Dosage des alcaloïdes de l'ergot Lebensmittel - Bestimmung von Ergotalkaloiden in
dans les céréales et les produits céréaliers par Getreiden und Getreideerzeugnissen mit dSPE-
purification par dSPE et CL-SM/SM Reinigung und LC-MS/MS
This European Standard was approved by CEN on 21 September 2020.
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
© 2021 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17425:2021 E
worldwide for CEN national Members.
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Principle . 6
5 Reagents . 6
6 Apparatus and equipment . 8
7 Procedure . 9
7.1 Preparation of the test sample . 9
7.2 Extraction of ergot alkaloids. 10
7.2.1 Precautions . 10
7.2.2 Test sample . 10
7.2.3 Spiking procedure . 10
7.2.4 Sample extraction . 10
7.2.5 Clean-up. 10
8 LC-MS/MS analysis . 10
8.1 General. 10
8.2 Batch composition and analytical sequence . 11
8.3 Identification . 11
8.4 Calibration . 11
9 Calculation . 11
9.1 Calculation of individual ergot alkaloid concentration . 11
9.2 Calculation of ergot alkaloid sum concentration . 12
10 Precision . 12
10.1 General. 12
10.2 Repeatability . 12
10.3 Reproducibility . 12
11 Test report . 13
Annex A (informative) Example conditions for suitable LC-MS/MS systems . 14
® ®
A.1 System settings for Waters Xevo TQ-S Mass Spectrometer. 14
A.1.1 Settings for chromatography . 14
A.1.2 Detector parameters. 15 ®
A.2 System settings for Micromass Quattro Ultima Pt Mass Spectrometer . 17
A.2.1 Settings for chromatography . 17
A.2.2 Detector parameters. 18
A.3 System settings for SCIEX API 5500 Mass Spectrometer . 19
A.3.1 Settings for chromatography . 19
A.3.2 Detector parameters . 20
A.4 Other LC conditions . 22
Annex B (informative) Typical chromatograms . 23
Annex C (informative) Precision data . 29
Annex D (informative) Data comparison . 43
Bibliography . 44
European foreword
This document (EN 17425:2021) has been prepared by Technical Committee CEN/TC 275 “Food
analysis - Horizontal methods”, the secretariat of which is held by DIN.
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 2021, and conflicting national standards shall
be withdrawn at the latest by October 2021.
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 mandate 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.
Introduction
Ergot alkaloids are a group of mycotoxins produced by several species of Claviceps fungi growing on
cereals and forage grass. These toxins are a risk for consumers as they can enter the food chain. All
ergot alkaloids share a common structure, the ergoline system, and are divided into several classes,
based on the presence of functional groups. The chiral carbon atom C-8 is responsible for the
epimerization.
The isomers of each of these compounds are nominally known as ‘ines’ and the ‘inines’. And besides,
ergocryptine and ergocryptinine can both occur as α- and β-forms.
WARNING 1 — Suitable precaution and protection measures need to be taken when carrying out
working steps with harmful chemicals. The latest version of the hazardous substances
ordinance, Regulation (EC) No 1907/2006 [3], should be taken into account as well as
appropriate national statements.
WARNING 2 — The use of this document can involve hazardous materials, operations and
equipment. This document does not purport to address all the safety problems associated with
its use. It is the responsibility of the user of this document to establish appropriate safety and
health practices and determine the applicability of regulatory limitations prior to use.
WARNING 3 — Ergot alkaloids can cause vasoconstrictive, neurotoxic, reproductive and
developmental adverse effects, and can be acutely and chronically toxic [4].
1 Scope
This document describes a method for the determination of the sum of six ergot alkaloids (ergocornine,
ergometrine, ergocristine, ergotamine, ergosine and ergocryptine) and their -inine epimer pairs by
liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) after clean-up by
dispersive solid phase extraction (dSPE).
The method has been validated in the range 13,2 µg/kg to 168 µg/kg for the sum of the twelve ergot
alkaloids, in rye flour, rye bread and cereal products (breakfast cereal, infant breakfast cereal, and
crispbread) that contained rye as an ingredient, as well as seeded wholemeal flour and a barley and rye
flour mixture.
Method performance was satisfactory in the range 24,1 µg/kg to 168 µg/kg, however at lower
concentrations RSD values were greater than 44 %, and HorRat values exceeded 2,0, indicating the
R
method may not be fully suitable at concentrations below 24 µg/kg for sum of ergot alkaloids, although
it is suitable for screening at these concentrations.
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, Water for analytical laboratory use - Specification and test methods (ISO 3696)
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: a
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