Reference materials and representative test materials: the nanotechnology case

  • G. RoebbenEmail author
  • K. Rasmussen
  • V. Kestens
  • T. P. J. Linsinger
  • H. Rauscher
  • H. Emons
  • H. Stamm


An increasing number of chemical, physical and biological tests are performed on manufactured nanomaterials for scientific and regulatory purposes. Existing test guidelines and measurement methods are not always directly applicable to or relevant for nanomaterials. Therefore, it is necessary to verify the use of the existing methods with nanomaterials, thereby identifying where modifications are needed, and where new methods need to be developed and validated. Efforts for verification, development and validation of methods as well as quality assurance of (routine) test results significantly benefit from the availability of suitable test and reference materials. This paper provides an overview of the existing types of reference materials and introduces a new class of test materials for which the term ‘representative test material’ is proposed. The three generic concepts of certified reference material, reference material (non-certified) and representative test material constitute a comprehensive system of benchmarks that can be used by all measurement and testing communities, regardless of their specific discipline. This paper illustrates this system with examples from the field of nanomaterials, including reference materials and representative test materials developed at the European Commission’s Joint Research Centre, in particular at the Institute for Reference Materials and Measurements (IRMM), and at the Institute for Health and Consumer Protection (IHCP).


Nanotechnology Nanomaterial Certified reference material Representative test material 



The authors wish to thank Juan Riego Sintes (JRC-IHCP) for valuable comments.


  1. Aschberger K, Micheletti C, Sokull-Klüttgen B, Christensen FM (2011) Analysis of currently available data for characterising the risk of engineered nanomaterials to the environment and human health—lessons learned from four case studies. Environ Int 37:1143–1156CrossRefGoogle Scholar
  2. Braun A, Franks K, Kestens V, Roebben G, Lamberty A, Linsinger TPJ (2011) Certification of equivalent spherical diameters of silica nanoparticles in water, certified reference material ERM®-FD100, EUR 24620 EN, ISBN 978-92-79-18676-9. Publications Office of the European Union, LuxembourgGoogle Scholar
  3. EC (European Commission) (1967) Council Directive 67/548/EEC of 27 June 1967 on the approximation of laws, regulations and administrative provisions relating to the classification, packaging and labelling of dangerous substances. OJ L 196, 16.8.1967, 1–98Google Scholar
  4. EC (European Commission) (2004) Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on the harmonisation of laws, regulations and administrative provisions relating to the application of the principles of good laboratory practice and the verification of their applications for tests on chemical substances. OJ L 50, 20.2.2004, p. 44–59Google Scholar
  5. EC (European Commission) (2006) Regulation (EC) No 1907/2006 of the European Parliament and of the Council of 18 December 2006 concerning the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), establishing a European Chemicals Agency, amending Directive 1999/45/EC and repealing Council Regulation (EEC) No 793/93 and Commission Regulation (EC) No 1488/94 as well as Council Directive 76/769/EEC and Commission Directives 91/155/EEC, 93/67/EEC, 93/105/EC and 2000/21/EC. OJ L 396, 30.12.2006, 1–849Google Scholar
  6. EC (European Commission) (2011) Commission Recommendation of 18 October 2011 on the definition of nanomaterial (2011/696/EU). OJ L 275, 20.10.2011, 38–40Google Scholar
  7. Emons H (2006) The ‘RM family’—identification of all its members. Accred Qual Assur 10:690–691CrossRefGoogle Scholar
  8. Emons H, Linsinger TPJ, Gawlik BM (2004) Reference materials: terminology and use. Can’t one see the forest for the trees? Trends Anal Chem 23:442–449CrossRefGoogle Scholar
  9. Franks K, Braun A, Charoud-Got J, Couteau O, Kestens V, Lamberty A, Linsinger TPJ, Roebben G (2012) Certification of the equivalent spherical diameters of silica nanoparticles in aqueous solution, certified reference material ERM®-FD304, EUR 25018 EN, ISBN 978-92-79-21867-5. Publications Office of the European Union, LuxembourgGoogle Scholar
  10. IRMM (Institute for Reference Materials and Measurements) (2005) IRMM-481 Product Information Sheet, December 2005,
  11. ISO (International Organization for Standardization) (2006) ISO Guide 35, reference materials—General and statistical principles for certification. ISO, GenevaGoogle Scholar
  12. ISO (International Organization for Standardization) (2008a) ISO Guide 30:1992/Amd.1:2008, Terms and definitions used in connection with reference materials. ISO, GenevaGoogle Scholar
  13. ISO (International Organization for Standardization) (2008b) ISO 148-2:2008 Metallic materials, Charpy pendulum impact test, Part 2: Verification of testing machines, ISO, GenevaGoogle Scholar
  14. ISO (International Organization for Standardization) (2009) ISO Guide 34. General requirements for the competence of reference material producers, ISO, GenevaGoogle Scholar
  15. ISO (International Organization for Standardization) (2010a) ISO/TS 80004–1:2010 Nanotechnologies—Vocabulary—Part 1: Core terms. ISO, GenevaGoogle Scholar
  16. ISO (International Organization for Standardization) (2010b) ISO 1182:2010 Reaction to fire tests for products. Non-combustibility test, ISO, GenevaGoogle Scholar
  17. ISO (International Organization for Standardization) (2011) ISO/DTR 13014, Nanotechnologies, Guidance on physico-chemical characterization for manufactured nano-objects submitted for toxicological testing, (approved for publication) ISO, GenevaGoogle Scholar
  18. ISO (International Organization for Standardization) (2012) ISO Concept Database,, accessed 11 January 2012
  19. IUPAC (International Union for Pure and Applied Chemistry) (1995) Harmonized guidelines for internal quality control in analytical chemistry laboratories. Pure Appl Chem 67:649–666CrossRefGoogle Scholar
  20. IUPAC (International Union for Pure and Applied Chemistry) (2012) IUPAC compendium of chemical terminology—the gold book, Accessed 11 January 2012
  21. JCGM (Joint Committee for Guides in Metrology) (2008). International vocabulary of metrology—Basic and general concepts and associated terms (VIM), 3rd Edition, JCGM 200:2008, Sevres, France, 2008.
  22. Johnston HJ, Hutchison GR, Christensen FM, Peters S, Hankin S, Stone V (2009) Identification of the mechanisms that drive the toxicity of TiO2 particulates; the contribution of physicochemical characteristics. Part Fibre Toxicol 6:33. doi: 10.1186/1743-8977-6-33 CrossRefGoogle Scholar
  23. Johnston HJ, Hutchison GR, Christensen FM, Peters S, Hankin S, Aschberger K, Stone V (2010a) A critical review of the biological mechanisms underlying the in vivo and in vitro toxicity of carbon nanotubes: the contribution of physico-chemical characteristics. Nanotoxicology 4:207–246CrossRefGoogle Scholar
  24. Johnston HJ, Hutchison GR, Christensen FM, Peters S, Hankin S, Stone V (2010b) A review of the in vivo and in vitro toxicity of silver and gold particulates: particle attributes and biological mechanisms responsible for the observed toxicity. Crit Rev Toxicol 40:328–346CrossRefGoogle Scholar
  25. Klein CL, Comero S, Stahlmecke B, Romazanov J, Kuhlbusch TAJ, Van Doren E, De Temmerman PJ, Mast J, Wick P, Krug H, Locoro G, Hund-Rinke K, Kördel W, Friedrichs S, Maier G, Werner J, Linsinger T and Gawlik BM. (2011). NM-Series of Representative Manufactured Nanomaterials, NM-300 Silver, Characterisation, Stability, Homogeneity. EUR 24693 EN. ISBN 978-92-79-19068-1Google Scholar
  26. Linsinger TPJ, Roebben G, Solans C, Ramsch R (2011) Reference materials for measuring the size of nanoparticles. Trends Anal Chem 30:18–27CrossRefGoogle Scholar
  27. NIST (National Institute for Standardisation) (2011) NIST releases First Certified Reference Material for Single-Wall Carbon Nanotubes,
  28. OECD (Organisation for Economic Co-operation and Development) (1998) OECD Principles on Good Laboratory Practice, in OECD Series on principles of Good Laboratory Practice and Compliance Monitoring, Number 1 ENV/MC/CHEM(98)17Google Scholar
  29. OECD (Organisation for Economic Co-operation and Development) (2005) Guidance Document on the Validation and International Acceptance of new or Updated Test Methods for Hazard Assessment, ENV/JM/MONO(2005)14, in OECD Series on Testing and Assessment, Number 34Google Scholar
  30. OECD (Organisation for Economic Co-operation and Development) (2008) OECD-WPMN, Series on the Safety of Manufactured Nanomaterials, Number 6, List of manufactured nanomaterials and list of endpoints for phase one of the OECD testing programme, ENV/JM/MONO[2008]13 (2008)Google Scholar
  31. OECD (Organisation for Economic Co-operation and Development) (2009) Preliminary Review of OECD Test Guidelines for their Applicability to Manufactured Nanomaterials. ENV/JM/MONO(2009)21Google Scholar
  32. OECD (Organisation for Economic Co-operation and Development) (2010) Guidance Manual for the Testing of Manufactured Nanomaterials: OECD’s Sponsorship Programme; first revision. ENV/JM/MONO(2009)20/REVGoogle Scholar
  33. Poland CA, Duffin R, Kinloch I, Maynard A, Wallace WAH, Seaton A, Stone V, Brown S, Macnee W, Donaldson K (2008) Carbon nanotubes introduced into the abdominal cavity of mice show asbestos-like pathogenicity in a pilot study. Nat Nanotechnol 3:423–428CrossRefGoogle Scholar
  34. Roebben G, Linsinger TPJ, Lamberty A, Emons H (2010) Metrological traceability of the measured values of properties of engineering materials. Metrologia 47:S23–S31CrossRefGoogle Scholar
  35. Roebben G, Emons H, Reiners G (2011), Nanoscale Reference Materials, in Nanotechnology Standards, eds. V. Murashov, J. Howard, Springer Science+Business Media, New York/Heidelberg, pp 53–75Google Scholar
  36. Scaravelli E, Brohée M, Marchelli R, van Hengel AJ (2009) The effect of heat treatment on the detection of peanut allergens as determined by ELISA and real-time PCR. Anal Bioanal Chem 395:127–137CrossRefGoogle Scholar
  37. Singh C, Friedrichs S, Levin M, Birkedal R, Jensen KA, Pojana G, Wohlleben W, Schulte S, Wiench K, Turney T, Koulaeva O, Marshall D, Hund-Rinke K, Kördel W, Van Doren E, De Temmerman PJ, Abi Daoud Francisco M, Mast J, Gibson N, Koeber R, Linsinger T, Klein CL. (2011), NM-Series of Representative Manufactured Nanomaterials, Zinc Oxide NM-110, NM-111, NM-112, NM-113, Characterisation and Test Item Preparation. EUR 25066 EN, ISBN 978-92-79-22215-3Google Scholar
  38. UNODC (United Nations Office on Drugs and Crime) (2009) Glossary of terms for quality assurance and good laboratory practices, ST/NAR/26/Rev.1Google Scholar
  39. Warheit DB (2010) Debunking some misconceptions about nanotoxicology. Nano Lett 10:4777–4782CrossRefGoogle Scholar
  40. WHO (World Health Organisation) International Biological reference Preparations, Geneva (2011), catalogue downloadable from

Copyright information

© European Union  2013

Authors and Affiliations

  • G. Roebben
    • 1
    Email author
  • K. Rasmussen
    • 2
  • V. Kestens
    • 1
  • T. P. J. Linsinger
    • 1
  • H. Rauscher
    • 2
  • H. Emons
    • 1
  • H. Stamm
    • 2
  1. 1.Institute for Reference Materials and MeasurementsJoint Research Centre of the European CommissionGeelBelgium
  2. 2.Institute for Health and Consumer ProtectionJoint Research Centre of the European CommissionIspraItaly

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