Animal feeding stuffs - Methods of sampling and analysis - Determination of pyrrolizidine alkaloids in animal feeding stuff by LC-MS/MS

This document specifies a method for the quantitative determination of pyrrolizidine alkaloids (PA) in the concentration ranges shown in Table 1 in complete and supplementary feed and in forages by liquid chromatography tandem mass spectrometry (LC-MS/MS) after solid phase extraction (SPE) clean-up.
Table 1 - Summary of concentration ranges per PA tested in the collaborative trial
NOTE 1   A second method was part of the method validation collaborative main trial. For this method PA-N-Oxides are reduced by adding zinc powder to the extract of the feed material. The following steps correspond to the first and main method. Quantitative results for each PA except the otonecine type PA senkirkine represent the sum of the free PA base and its corresponding N-oxide.
NOTE 2   Due to insufficient numbers of data for some analyte-matrix combinations statistical evaluation was not valid for standardization. Received data indicated the methods applicability in experienced laboratories with appropriate quality assurance measures. Therefore, the method description is included as an informative annex (Annex D).

Futtermittel - Probenahme- und Untersuchungsverfahren - Bestimmung von Pyrrolizidinalkaloiden in Futtermitteln mittels LC-MS/MS

Dieses Dokument legt ein Verfahren zur quantitativen Bestimmung von Pyrrolizidinalkaloiden (PA) in den in Tabelle 1 angegebenen Konzentrationsbereichen in Allein- und Ergänzungsfuttermitteln sowie in Grünfutter mittels Flüssigchromatographie mit Massenspektrometrie-Kopplung (LC MS/MS) nach Aufreinigung durch Festphasenextraktion (SPE, en: solid phase extraction) fest.
Tabelle 1 - Zusammenfassung der im Ringversuch geprüften Konzentrationsbereiche je PA
ANMERKUNG 1   Ein zweites Verfahren war Teil des Haupt-Ringversuchs zur Verfahrensvalidierung. Bei diesem Verfahren werden PA N Oxide durch Zugabe von Zinkpulver zum Extrakt des Futtermittels reduziert. Die folgenden Schritte entsprechen dem ersten und wichtigsten Verfahren. Die quantitativen Ergebnisse für jedes PA mit Ausnahme des PA vom Otonecin-Typ Senkirkin stellen die Summe der freien PA-Base und ihres entsprechenden N Oxids dar.
ANMERKUNG 2   Aufgrund einer unzureichenden Anzahl von Daten für einige Analyt-Matrix-Kombinationen war die statistische Auswertung für die Normung nicht gültig. Die erhaltenen Daten weisen auf die Anwendbarkeit des Verfahrens in erfahrenen Laboren mit entsprechenden Qualitätssicherungsmaßnahmen hin. Daher ist die Verfahrensbeschreibung als informativer Anhang (Anhang D) beigefügt.

Alimentation animale - Méthodes d’échantillonnage et d’analyse - Dosage des alcaloïdes pyrrolizidiniques dans l'alimentation animale par CL-SM/SM

Le présent document spécifie une méthode de dosage des alcaloïdes pyrrolizidiniques (AP) dans les gammes de concentrations indiquées dans le Tableau 1 dans les aliments composés pour animaux complets et complémentaires ainsi que dans les fourrages, par chromatographie liquide couplée à une spectrométrie de masse en tandem (CL-SM/SM) après purification par extraction en phase solide (SPE).
Tableau 1 - Résumé des gammes de concentrations par AP soumis à essai lors de l’essai interlaboratoires
NOTE 1   Une seconde méthode faisait partie de l’essai interlaboratoires principal de validation de la méthode. Pour cette méthode, les N-oxydes d‘AP sont réduits en ajoutant de la poudre de zinc à l’extrait d’aliment composé pour animaux. Les étapes suivantes correspondent à la première et principale méthode. Les résultats quantitatifs pour chaque AP, excepté l’AP senkirkine de type otonécine, représentent la somme de la base libre d’AP et de son N-oxyde correspondant.
NOTE 2   En raison du nombre insuffisant de données pour certaines combinaisons analyte-matrice, l’évaluation statistique n’était pas valide pour la normalisation. Les données reçues ont indiqué l’applicabilité des méthodes au sein de laboratoires expérimentés avec des mesures d’assurance qualité appropriées. Par conséquent, la description de la méthode est incluse à titre d’annexe informative (Annexe D).

Krma - Metode vzorčenja in analize - Določanje pirolizidinskih alkaloidov v krmi z LCMS/MS

Ta dokument opisuje metodo za kvantitativno določevanje pirolizidinskih alkaloidov (PA) v polnovredni in dopolnilni krmi s tandemsko masno spektrometrijo s tekočinsko kromatografijo (LC-MS/MS) po izpiranju z ekstrakcijo na trdni fazi (SPE).
Ta metoda je bila uspešno potrjena v primerjalnem preskusu za matrice s polnovredno krmo za konje, dopolnilno krmo za konje, dopolnilno krmo za glodavce, seno, lucerno in travno silažo. Potrditev je bila izvedena za pirolizidinske alkaloide (PA) in vrednosti koncentracije, navedene v tabeli 1. Dokazano je bilo, da izomerna para pirolizidinskih alkaloidov sineciverin in senecionin ter senecivernin-N-oksid in senecionin-N-oksid, ni mogoče določiti posamezno zaradi neustreznega kromatografskega ločevanja. Vseeno pa so bile vsote posameznih pirolizidinskih alkaloidov izomernih parov kvantificirane z ustrezno stopnjo ponovljivosti. Upoštevati je treba sočasno eluacijo drugih izomerov pirolizidinskih alkaloidov, ki niso zajeti v metodo. Seznam izomerov s potencialno sočasno eluacijo je podan v dodatku E.
Čeprav je kalibracijsko območje protokola metode določeno od 10 μg/kg do 300 μg/kg, so rezultati primerjalne študije pokazali, da redčenje ekstraktov vzorcev z ekstrakti slepih vzorcev omogoča kvantitativno določanje koncentracij, ki presegajo kalibracijsko območje. Zadovoljiva ponovljivost je vila dosežena pri kvantificiranju do 1428 μg/kg za posamezen pirolizidinski alkaloid in do 887 μg/kg za vsoto izomernih parov.
OPOMBA 1: Druga metoda je bila del primerjalnega glavnega preskusa potrjevanja metode. Pri tej metodi so bili PA-N-oksidi zmanjšani z dodajanjem cinkovega prahu ekstraktu krmnega material. Spodnji koraki ustrezajo prvi in glavni metodi. Kvantitativni rezultati za vsak pirolizidinski alkaloid razen pirolizidinskega alkaloida senkirkin vrste otenocin predstavljajo vsoto proste baze pirolizidinskih alkaloidov in njen ustrezen N-oksid.
OPOMBA 2: Zaradi nezadostnega števila podatkov za nekatere kombinacije analita in matrike statistična ocena za standardizacijo ni bila veljavna. Pridobljeni rezultati so kazali uporabnost metod v izkušenih laboratorijih z ustrezno kakovostjo ukrepov za zagotavljanje kakovosti. Za to je opis metode vključen kot informativni dodatek (dodatek D).

General Information

Status
Published
Publication Date
28-Feb-2023
Current Stage
6060 - Definitive text made available (DAV) - Publishing
Start Date
01-Mar-2023
Due Date
02-Dec-2020
Completion Date
01-Mar-2023

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Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN 17683:2023
01-maj-2023
Krma - Metode vzorčenja in analize - Določanje pirolizidinskih alkaloidov v krmi z
LCMS/MS
Animal feeding stuffs - Methods of sampling and analysis - Determination of pyrrolizidine
alkaloids in animal feeding stuff by LC-MS/MS
Futtermittel - Probenahme- und Untersuchungsverfahren - Bestimmung von
Pyrrolizidinalkaloiden in Futtermitteln mittels LC-MS/MS
Alimentation animale - Méthodes d’échantillonnage et d’analyse - Dosage des alcaloïdes
pyrrolizidiniques dans l'alimentation animale par CL-SM/SM
Ta slovenski standard je istoveten z: EN 17683:2023
ICS:
65.120 Krmila Animal feeding stuffs
SIST EN 17683:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
SIST EN 17683:2023

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SIST EN 17683:2023


EN 17683
EUROPEAN STANDARD

NORME EUROPÉENNE

March 2023
EUROPÄISCHE NORM
ICS 65.120
English Version

Animal feeding stuffs - Methods of sampling and analysis -
Determination of pyrrolizidine alkaloids in animal feeding
stuff by LC-MS/MS
Alimentation animale - Méthodes d'échantillonnage et Futtermittel - Probenahme- und
d'analyse - Dosage des alcaloïdes pyrrolizidiniques Untersuchungsverfahren - Bestimmung von
dans l'alimentation animale par CL-SM/SM Pyrrolizidinalkaloiden in Futtermitteln mittels LC-
MS/MS
This European Standard was approved by CEN on 17 January 2023.

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, Türkiye 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
© 2023 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17683:2023 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
SIST EN 17683:2023
EN 17683:2023 (E)
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 7
3 Terms and definitions . 7
4 Principle . 7
4.1 General. 7
4.2 Reagents . 7
4.3 Analytical standards . 7
4.4 Chemicals . 8
4.5 Solutions . 9
4.5.1 Extraction solution 0,05 mol/l aqueous sulphuric acid . 9
4.5.2 Aqueous ammoniacal solution (20/80
...

SLOVENSKI STANDARD
oSIST prEN 17683:2021
01-september-2021
Krma - Metode vzorčenja in analize - Določanje pirolizidinskih alkaloidov v krmi z
LCMS/MS
Animal feeding stuffs- Methods of sampling and analysis - Determination of pyrrolizidine
alkaloids in animal feeding stuff by LCMS/MS
Futtermittel - Probenahme- und Untersuchungsverfahren - Bestimmung von
Pyrrolizidinalkaloide in Futtermitteln mittels LC-MS/MS
Aliments pour animaux : Méthodes d’échantillonnage et d’analyse — Dosage des
alcaloïdes pyrrolizidiniques dans les aliments pour animaux par CL-SM/SM
Ta slovenski standard je istoveten z: prEN 17683
ICS:
65.120 Krmila Animal feeding stuffs
oSIST prEN 17683:2021 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
oSIST prEN 17683:2021

---------------------- Page: 2 ----------------------
oSIST prEN 17683:2021


DRAFT
EUROPEAN STANDARD
prEN 17683
NORME EUROPÉENNE

EUROPÄISCHE NORM

July 2021
ICS 65.120
English Version

Animal feeding stuffs: Methods of sampling and analysis -
Determination of pyrrolizidine alkaloids in animal feeding
stuff by LCMS/MS
 Futtermittel - Probenahme- und
Untersuchungsverfahren - Bestimmung von
Pyrrolizidinalkaloide in Futtermitteln mittels LC-
MS/MS; Deutsche und Englische Fassung prEN WI
00327113:2018
This draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 327.

If this draft becomes a European Standard, 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.

This draft European Standard was established by CEN 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.

Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.

Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.


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. prEN 17683:2021 E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------
oSIST prEN 17683:2021
prEN 17683:2021 (E)
Contents Page
European foreword . 3
Introduction . 4
1 Scope . 5
2 Normative references . 7
3 Terms and definitions . 7
4 Principle . 7
4.1 General. 7
4.2 Reagents . 7
4.3 Analytical standards . 7
4.4 Chemicals . 8
4.5 Solutions .
...

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