EN 16418:2014

Paper and board - Determination of the cytotoxicity of aqueous extracts using a metabolically competent hepatoma cell line (HepG2)

This European Standard specifies a test method for the laboratory assessment of the potential cytotoxic effect of paper and board intended to come into contact with foodstuffs using specifically the HepG2 cell line.
Compared to the EN 15845[1], HepG2 cells are more representative of a human oral exposure to xenobiotics, due to the presence in the cells of phase I, II and III enzymes of the metabolism.

Papier und Pappe - Bestimmung der Zytotoxizität von wässrigen Extrakten unter Verwendung einer metabolisch kompetenten Hepatom-Zelllinie (HepG2)

Dieses Dokument legt ein Prüfverfahren für die Laboruntersuchung des potentiellen zytotoxischen Effekts von Lebensmittelkontaktmaterialien aus Papier oder Pappe, spezifisch unter Verwendung der HepG2-Zelllinie, fest.
Im Vergleich zur EN 15845 [1] repräsentieren HepG2-Zellen besser den oralen Kontakt des Menschen mit Xenobiotika aufgrund des Vorhandenseins von Phase I, II und III –Stoffwechselenzymen in den Zellen.

Papier et carton - Détermination de l’effet cytotoxique d’extraits aqueux en utilisant une lignée cellulaire d’hépatome possédant des enzymes du métabolisme (cellules HepG2)

La présente Norme européenne définit une méthode d’essai utilisant spécifiquement la lignée cellulaire HepG2 pour l’évaluation en laboratoire de l’effet cytotoxique potentiel du papier et du carton destinés à entrer en contact avec des denrées alimentaires.
Par rapport à la méthode décrite dans l’EN 15845[1], les cellules HepG2 sont plus représentatives de l’exposition des individus aux xénobiotiques par voie orale, en raison de la présence dans les cellules d’enzymes du métabolisme de phase I, II et III.

Papir, karton in lepenka - Ugotavljanje citotoksičnosti vodnih ekstraktov z uporabo metabolno kompetentnih hepatom celic linije (HepG2)

Standard EN 16418 določa preskusno metodo za laboratorijsko oceno morebitnega citotoksičnega učinka materialov iz papirja, kartona in lepenke, ki imajo stik z živili, z uporabo zlasti celic linije HepG2. V primerjavi s standardom EN 15845 celice HepG2 bolje predstavljajo človeško oralno izpostavljenost ksenobiotikom zaradi prisotnosti v celicah encimov faz I, II in III metabolizma.

General Information

• Status: Published
• Publication Date: 01-Apr-2014
• Withdrawal Date: 30-Oct-2014
• ICS: 85.060 - Paper and board
• Technical Committee: CEN/TC 172 - Pulp, paper and board
• Drafting Committee: CEN/TC 172/WG 12 - Cytotoxicity
• Current Stage: 9093 - Decision to confirm - Review Enquiry
• Completion Date: 16-Mar-2020
• Ref Project: SIST EN 16418:2014 - Paper and board - Determination of the cytotoxicity of aqueous extracts using a metabolically competent hepatoma cell line (HepG2)

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Papier und Pappe - Bestimmung der Zytotoxizität von wässrigen Extrakten
unter Vewendung einer metabolisch kompetenten Hepatom-Zelllinie (HepG2)Papier et carton - Détermination de l'effet cytotoxique d'extraits aqueux en utilisant une lignée cellulaire d'hépatome possédant des enzymes du métabolisme (cellules HepG2)Paper and board - Determination of the cytotoxicity of aqueous extracts using a metabolically competent hepatoma cell line (HepG2)85.060Papir, karton in lepenkaPaper and boardICS:Ta slovenski standard je istoveten z:EN 16418:2014SIST EN 16418:2014en,fr,de01-september-2014SIST EN 16418:2014SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16418
April 2014 ICS 85.060 English Version
Paper and board - Determination of the cytotoxicity of aqueous extracts using a metabolically competent hepatoma cell line (HepG2)
Papier et carton - Détermination de l'effet cytotoxique d'extraits aqueux en utilisant une lignée cellulaire d'hépatome possédant des enzymes du métabolisme (cellules HepG2)
Papier und Pappe - Bestimmung der Zytotoxizität von wässrigen Extrakten unter Verwendung einer metabolisch kompetenten Hepatom-Zelllinie (HepG2) This European Standard was approved by CEN on 8 February 2014.
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, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
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B-1000 Brussels © 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16418:2014 ESIST EN 16418:2014

Foreword .3 1 Scope .4 2 Normative references .4 3 Terms and definitions .4 4 Principle .5 5 Reagents .5 6 Cell line .7 6.1 Generating the cell strain .7 6.2 Maintaining the cell strain .7 6.3 Storing the cell strain .8 7 Food simulants used for testing .8 7.1 Reference water (3.1) .8 8 Cleaning laboratory glassware .8 8.1 Cleaning liquids for laboratory glassware .8 8.2 Cleaning procedure for laboratory glassware .8 9 Equipment .9 9.1 Equipment for the migration test .9 9.2 Cell culture equipment .9 9.3 Equipment used for cytotoxicity testing .9 10 Preparation of specimens . 10 10.1 General . 10 10.2 Paper and board intended for wet contact . 10 11 Cytotoxicity assessment . 10 11.1 Principle . 10 11.2 General . 10 11.3 Cell seeding . 11 11.4 Preparation of samples . 11 11.5 Cell culture treatment . 11 11.6 Preparation of the chromatography sheet . 12 11.7 Kinetics of uridine incorporation in the cell RNA . 12 11.8 Measurements of the RNA synthesis . 12 12 Expression of the results . 13 12.1 Graphic representation of the results . 13 12.2 Calculation of percentage RNA synthesis and the validity of the test . 14 13 Interpretation of the results . 15 13.1 Results for the reference sample . 15 13.2 Results of the positive control sample . 15 13.3 Results for the test sample . 15 14 Precision . 15 15 Test report . 15 Annex A (informative)
96-well plates configuration . 17 Annex B (informative)
RNA Synthesis rate inhibition cytotoxicity test work flow . 18 Annex C (informative)
Validation of the two methods (option A and B) . 19 Bibliography . 20 SIST EN 16418:2014
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