EN 16215:2020
(Main)Animal feeding stuffs: Methods of sampling and analysis - Determination of dioxins and dioxin-like PCBs and of indicator PCBs by GC/HRMS
Animal feeding stuffs: Methods of sampling and analysis - Determination of dioxins and dioxin-like PCBs and of indicator PCBs by GC/HRMS
This document is applicable to the determination of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), (together termed ‘dioxins’ (PCDD/Fs)) and dioxin-like PCBs and non-dioxin-like PCBs (dl-PCBs and ndl-PCBs) in animal feeding stuffs. Collaborative studies have been carried out. The method is suitable for the determination of dioxins, dl-PCBs and ndl-PCBs at the appropriate MRL in compound feed and ingredients e.g. oil, mineral clay. The method is applicable to samples containing trace level amounts of one or more dioxins, dioxin-like PCBs and non-dioxin-likePCBs. The limit of quantification (LOQ) is
- 0,05 pg/g (OCDD/F = 0,1 pg/g) for the relevant individual congeners of dioxins/furans,
- 0,05 pg/g for non-ortho PCBs,
- 10 pg/g for mono-ortho PCBs, and
- 100 pg/g for non-dioxin-like-PCBs.
For determination of dioxins and dioxin-like PCBs, the procedure can be used as confirmatory method as defined by Commission Regulation (EC) No 152/2009 for dioxins and dl-PCB in feed [1]. Confirmatory methods as described in this standard are high-resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) methods. If only the analysis of non-dioxin-like PCBs is required, a GC-LRMS method can be used (e.g. EN 15741 [2]) provided that appropriate analytical performance criteria are met in the relevant range for the matrix of interest.
This document is split into four modules. Each module describes a part of the whole procedure (see Figure 1 and Figure 2) to be followed:
a) Module A: Description of standards which might be used;
b) Module B: Description of extraction procedures;
c) Module C: Description of clean-up procedures;
d) Module D: GC/HRMS determination.
Each module describes a part of the whole method as well as, when applicable, alternatives which should be equivalent. Each module has to be regarded as an example. Combining modules and/or alternatives gives a highly flexible, "performance based" procedure. It is permitted to modify the method if all performance criteria laid down in Commission Regulation (EC) No 152/2009 [1] are met.
Any deviation of the described method, combination of modules needs to be recorded as part of the QA/QC procedures of accredited laboratories and should be available on request.
Figure 1 - Flow scheme for the determination of dioxins, dl-PCBs and non-dioxin-like-PCBs in feed
Figure 2 - Flow scheme for the determination of dioxins, dl-PCBs and non-dioxin-like-PCBs in oil and fat
Futtermittel: Probenahme- und Untersuchungsverfahren - Bestimmung von Dioxinen und dioxin-ähnlichen PCB mittels GC/HRMS und von Indikator-PCB mittels GC/HRMS
Dieses Dokument ist anwendbar für die Bestimmung von polychlorierten Dibenzo p dioxinen (PCDD), polychlorierten Dibenzofuranen (PCDF), (zusammen als „Dioxine“ bezeichnet [PCDD/F]) und dioxin-ähnlichen PCB und nicht dioxin ähnlichen PCB (dl PCB und ndl PCB) in Futtermitteln. Ringversuche sind durchgeführt worden. Das Verfahren ist für die Bestimmung von Dioxinen, dl PCB und ndl PCB bei der entsprechenden MRL in Mischfutter und Inhaltsstoffen, z. B. Öl, Tonmineral, geeignet. Das Verfahren ist auf Proben anwendbar, die Spurenkonzentrationen von einem Dioxin oder mehreren Dioxinen, dioxin-ähnlichen PCB und nicht dioxin ähnlichen PCB enthalten. Die Bestimmungsgrenze (LOQ) beträgt
- 0,05 pg/g (OCDD/F = 0,1 pg/g) für die relevanten einzelnen Kongeneren der Dioxine/Furane,
- 0,05 pg/g für non-ortho PCB,
- 10 pg/g für mono-ortho PCB und
- 100 pg/g für nicht dioxin ähnliche PCB.
Für die Bestimmung von Dioxinen und dioxin-ähnlichen PCB kann das Verfahren, wie in der Verordnung (EG) Nr. 152/2009 der Kommission [1] für Dioxine und dl PCB in Futtermitteln festgelegt, als Bestätigungsverfahren angewendet werden. Bestätigungsverfahren, wie in dieser Norm beschrieben, sind Verfahren mit hochauflösender Gaschromatographie/hochauflösender Massenspektrometrie (HRGC/HRMS). Falls nur die Analyse der nicht dioxin ähnlichen PCB erforderlich ist, kann ein GC LRMS Verfahren (z. B. EN 15741 [2]) angewendet werden, vorausgesetzt, dass für die zu untersuchende Matrix im relevanten Bereich die entsprechenden analytischen Leistungskriterien erfüllt werden.
Dieses Dokument ist in vier Module aufgeteilt. Jedes Modul beschreibt einen Teil des gesamten Verfahrens (siehe Bild 1 und Bild 2) wie folgt:
a) Modul A: Beschreibung der Normen, die angewendet werden können;
b) Modul B: Beschreibung der Extraktionsverfahren;
c) Modul C: Beschreibung der Reinigungsverfahren;
d) Modul D: GC/HRMS Bestimmung.
Jedes Modul beschreibt einen Teil des gesamten Verfahrens sowie, falls anwendbar, Alternativen, die gleich¬wertig sein sollten. Jedes Modul muss als ein Beispiel betrachtet werden. Das Kombinieren von Modulen und/oder Alternativen ermöglicht ein hochflexibles, „leistungsbezogenes“ Verfahren. Es ist zulässig, das Verfahren zu verändern, wenn sämtliche Leistungskriterien entsprechend der Verordnung (EG) Nr. 152/2009 der Kommission [1] erfüllt werden.
Jede Abweichung vom beschriebenen Verfahren und die Kombination der Module müssen als Teil der QA/QC Verfahren von anerkannten Laboratorien aufgezeichnet werden und sollten auf Anfrage verfügbar sein.
Aliments des animaux : Méthodes d’échantillonnage et d’analyse - Dosage des dioxines, des PCB de type dioxine et des PCB indicateurs par GC/HRMS
Le présent document concerne le dosage des dibenzo-p-dioxines polychlorés (PCDD), des dibenzofuranes polychlorés (PCDF) (conjointement appelés « dioxines » (PCDD/F)) ainsi que des PCB de type dioxine et des PCB autres que ceux de type dioxine (dl-PCB et ndl-PCB) dans les aliments des animaux. Des études comparatives ont été menées. La méthode convient au dosage des dioxines, des dl PCB et des ndl-PCB à la LMR appropriée dans les aliments et les ingrédients composés tels que l’huile et l’argile minérale. La méthode est applicable aux échantillons contenant, à l’état de traces, un(e) ou plusieurs dioxines, PCB de type dioxine et PCB autres que ceux de type dioxine. La limite de quantification (LQ) est de :
- 0,05 pg/g (OCDD/F = 0,1 pg/g) pour les congénères individuels concernés des dioxines/furanes ;
- 0,05 pg/g pour les PCB non-ortho ;
- 10 pg/g pour les PCB mono-ortho ; et
- 100 pg/g pour les PCB autres que ceux de type dioxine.
Pour le dosage des dioxines et des PCB de type dioxine, le mode opératoire peut être utilisé comme méthode de confirmation, conformément aux définitions du Règlement (CE) no 152/2009 de la Commission relatif aux dioxines et aux dl-PCB dans les aliments [1]. Les méthodes de confirmation décrites dans la présente norme sont les méthodes de chromatographie en phase gazeuse haute résolution/spectrométrie de masse haute résolution (HRGC/HRMS). Si seule l’analyse des PCB autres que ceux de type dioxine est requise, une méthode GC-LRMS peut être utilisée (par exemple, EN 15741 [2]), à condition que les critères de performance analytique appropriés soient atteints dans la gamme de concentration correspondant aux aliments concernés.
Le présent document se divise en quatre modules. Chaque module décrit une partie de l’ensemble du mode opératoire (voir la Figure 1 et la Figure 2) à respecter :
a) Module A : Description des normes susceptibles d’être utilisées ;
b) Module B : Description des modes opératoires d’extraction ;
c) Module C : Description des modes opératoires de purification ;
d) Module D : Dosage par GC/HRMS.
Chaque module décrit une partie de l’ensemble de la méthode ainsi que, le cas échéant, des alternatives équivalentes. Chaque module doit être considéré comme un exemple. La combinaison des modules et/ou alternatives donne un mode opératoire très souple qui est « basé sur les performances ». Il est possible de modifier la méthode si tous les critères de performance énoncés dans le Règlement (CE) no 152/2009 de la Commission [1] sont remplis.
En cas d’écart par rapport à la méthode décrite, il est nécessaire d’enregistrer la combinaison de modules dans le cadre des procédures AQ/CQ des laboratoires accrédités et il convient que cette combinaison soit disponible sur demande.
[sans figure]
Figure 1 - Logigramme relatif au dosage des dioxines, des dl-PCB et des PCB autres que ceux de type dioxine dans les aliments
[sans figure]
Figure 2 - Logigramme relatif au dosage des dioxines, des dl-PCB et des PCB autres que ceux de type dioxine dans l’huile/la graisse
Krma: metode vzorčenja in analize - Določevanje dioksinov in dioksinu podobnih PCB z GC/HRMS in indikatorjev PCB z GC/HRMS
Ta dokument se uporablja za določevanje polikloriranih dibenzo-p-dioksinov (PCDD), polikloriranih dibenzofuranov (PCDF) (s skupnim imenom »dioksini« (PCDD/F)) ter dioksinom podobnih polikloriranih bifenilov in dioksinom nepodobnih polikloriranih bifenilov (dl-PCB in ndl-PCB) v krmi. Izvedene so bile medlaboratorijske študije. Metoda je primerna za določevanje dioksinov, dioksinom podobnih polikloriranih bifenilov in dioksinom nepodobnih polikloriranih bifenilov v krmi in sestavinah, na primer olju ter mineralni glini, pri ustrezni mejni vrednosti ostankov. Metoda se uporablja za vzorce, ki vsebujejo enega ali več dioksinov, dioksinom podobne poliklorirane bifenile in dioksinom nepodobne poliklorirane bifenile v sledovih. Mejne vrednosti kvantifikacije (LOQ) so:
– 0,05 pg/g (OCDD/F = 0,1 pg/g) za ustrezne posamezne analoge dioksinov/furanov,
– 0,05 pg/g za neorto poliklorirane bifenile,
– 10 pg/g za za monoorto poliklorirane bifenile in
– 100 pg/g za dioksinom nepodobne poliklorirane bifenile.
Pri določevanju dioksinov in dioksinom podobnih polikloriranih bifenilov je postopke mogoče uporabiti kot potrditveno metodo v skladu z Uredbo Komisije (ES) št. 152/2009 za dioksine in dioksinom podobne poliklorirane bifenile v krmi [1]. Potrditvene metode, kot je opisano v tem standardu, so metode visokoločljivostne plinske kromatografije/visokoločljivostne masne spektrometrije (HRGC/HRMS). Če je potrebna samo analiza dioksinom nepodobnih polikloriranih bifenilov, je mogoče uporabiti metodo plinske kromatografije/nizkoločljivostne masne spektrometrije (GC-LRMS) (npr. EN 15741 [2]) pod pogojem, da so za zadevno matriko v ustreznem območju izpolnjena ustrezna merila analitskega izvajanja.
Ta dokument je razdeljen v štiri module. Vsak modul opisuje del celotnega postopka (glej sliki 1 in 2), ki ga je treba upoštevati:
a) Modul A: opis standardov, ki se lahko uporabljajo;
b) Modul B: opis postopkov za ekstrakcijo;
c) Modul C: opis postopkov za očiščenje;
d) Modul D: določitev s plinsko kromatografijo/visokoločljivostno masno spektrometrijo.
Vsak modul opisuje del celotne metode in, kadar je to primerno, alternative, ki naj bodo enakovredne. Posamezni modul je treba obravnavati kot primer. S kombinacijo modulov in/ali alternativ se oblikuje izredno prilagodljiv postopek, ki temelji na učinkovitosti. Metodo je mogoče spremeniti, če so izpolnjena vsa merila učinkovitosti, ki so navedena v Uredbi Komisije (ES) št. 152/2009 [1]. V primeru odstopanj od opisane metode je treba kombinacijo modulov zabeležiti kot del postopkov za zagotavljanje in nadzor kakovosti (QA/QC) akreditiranih laboratorijev, ki naj bo na voljo za zahtevo.
Slika 1 – Shema poteka za določevanje dioksinov, dioksinom podobnih polikloriranih bifenilov in dioksinom nepodobnih polikloriranih bifenilov v krmi
Slika 2 – Shema poteka za določevanje dioksinov, dioksinom podobnih polikloriranih bifenilov in dioksinom nepodobnih polikloriranih bifenilov v olju in maščobi
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-marec-2020
Nadomešča:
SIST EN 16215:2012
Krma: metode vzorčenja in analize - Določevanje dioksinov in dioksinu podobnih
PCB z GC/HRMS in indikatorjev PCB z GC/HRMS
Animal feeding stuffs: Methods of sampling and analysis - Determination of dioxins and
dioxin-like PCBs by GC/HRMS and of indicator PCBs by GC/HRMS
Futtermittel - Probenahme- und Untersuchungsverfahren - Bestimmung von Dioxinen
und dioxin-ähnlichen PCBs mittels GC/HRMS und von Indikator-PCBs mittels GC/HRMS
Aliments des animaux : Méthodes d’échantillonnage et d’analyse - Dosage des dioxines,
des PCB de type dioxine et des PCB indicateurs par GC/HRMS
Ta slovenski standard je istoveten z: EN 16215:2020
ICS:
65.120 Krmila Animal feeding stuffs
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 16215
EUROPEAN STANDARD
NORME EUROPÉENNE
January 2020
EUROPÄISCHE NORM
ICS 65.120 Supersedes EN 16215:2012
English Version
Animal feeding stuffs: Methods of sampling and analysis -
Determination of dioxins and dioxin-like PCBs by
GC/HRMS and of indicator PCBs by GC/HRMS
Aliments des animaux : Méthodes d'échantillonnage et Futtermittel - Probenahme- und
d'analyse - Dosage des dioxines, des PCB de type Untersuchungsverfahren - Bestimmung von Dioxinen
dioxine et des PCB indicateurs par GC/HRMS und dioxin-ähnlichen PCBs mittels GC/HRMS und von
Indikator-PCBs mittels GC/HRMS
This European Standard was approved by CEN on 11 November 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
© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 16215:2020 E
worldwide for CEN national Members.
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 8
3 Terms and definitions . 8
4 Principle . 9
5 Reagents . 10
5.1 General . 10
5.2 Dioxins, furans, non-ortho PCBs, mono-ortho PCBs and non-dioxin-like PCBs and
their labelled analogues . 10
6 Apparatus . 10
7 Sampling . 10
8 Preparation of test sample . 11
9 Procedure. 11
9.1 General . 11
9.2 Animal feed stuff sample and oil/fat sample . 11
10 Extraction . 11
10.1 General . 11
10.2 Module BI Extraction using automated Pressurized Fluid Extraction (PFE) system . 12
10.3 Module BII: Manual extraction procedure . 13
11 Clean-up . 14
11.1 General . 14
11.2 Module CI: Automated clean-up . 16
11.3 Module CII: Manual sample clean-up, removal of fat and group separation . 19
11.4 Additional clean-up-steps for fraction A and fraction B of module CII . 22
11.5 Module CIII: Removal of matrix, manual sample clean-up and group separation . 24
12 Module D: Gas chromatograph-high resolution mass spectrometer (GC-HRMS) . 29
12.1 General . 29
12.2 Reagents and materials . 29
12.3 Procedure. 29
13 Calculation and expression of results . 31
13.1 General . 31
13.2 Calibration by Isotope Dilution . 31
13.3 Linearity . 31
13.4 Calibration criteria . 31
13.5 Identification and confirmation. 31
13.6 Calculation . 32
13.7 Toxic Equivalent (TEQ) values . 33
13.8 Recovery . 36
14 Precision . 36
15 Test report . 38
Annex A (informative) Description of PTV injection system . 39
A.1 Large volume injection (100 µl) – GC-HRMS Calibration mixtures dioxins and non-
ortho PCBs . 39
A.2 Sample preparation procedure . 40
A.3 PTV injection conditions using 100 µl injection . 40
Annex B (informative) Description of standards and concentration of the standard
solutions . 41
B.1 CAS Registry numbers for dioxins, furans, non-ortho PCBs, mono-ortho PCBs and
non-dioxin-like PCBs and their labelled analogues . 41
B.2 Standard solutions . 42
B.3 GC-standard solutions . 45
Annex C (informative) Example of automated procedure . 48
Annex D (informative) Mass spectrometer (MS) . 50
Annex E (informative) Use of additional clean-up after fractionation using a small multi-
layer silica column . 54
E.1 Preparation of the silica mixtures . 54
E.2 Preparation of the small multi-layer column . 54
Bibliography . 55
European foreword
This document (EN 16215:2020) 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 July 2020, and conflicting national standards shall be
withdrawn at the latest by July 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 supersedes EN 16215:2012.
In comparison with the previous edition, the following technical modifications have been made:
— incorrect technical details have been corrected,
— references have been updated,
— inconsistencies were removed, and
— editorial adaptations have been made.
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
The previous version of this document was developed in response to Directive 2002/32/EC of the
European Parliament and of the Council of 7 May 2002 on undesirable substances in animal feed. The
document provides analytical laboratories active in the field of feed analysis with guidance for the
analysis of dioxins and PCBs and meets criteria as set in Commission Regulation (EC) No 152/2009 of 27
January 2009 laying down the methods of sampling and analysis for the official control of feed.
In this updated version, obvious mistakes were corrected.
WARNING — The use of this document can involve hazardous materials, operations and
equipment. This standard does not purport to address all the safety problems associated with its
use. It is the responsibility of the user of this European Standard to establish appropriate safety
and health practices and determine the applicability of regulatory limitations prior to use.
1 Scope
This document is applicable to the determination of polychlorinated dibenzo-p-dioxins (PCDDs),
p
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