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Eprints ID : 16573
To link to this article : DOI:10.1002/anie.201403335
URL : http://dx.doi.org/10.1002/anie.201403335
To cite this version : Wick, Peter and Louw-Gaume, Anna E. and
Kucki, Melanie and Krug, Harald F. and Kostarelos, Kostas and
Fadeel, Bengt and Dawson, Kenneth A. and Salvati, Anna and
Vázquez, Ester and Ballerini, Laura and Tretiach, Mauro and
Benfenati, Fabio and Flahaut, Emmanuel and Gauthier, Laury and
Prato, Maurizio and Bianco, Alberto Classification Framework for
Graphene-Based Materials. (2014) Angewandte Chemie International
Edition, vol. 53 (n° 30). pp. 7714-7718. ISSN 1433-7851
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DOI: 10.1002/anie.201403335
Classification Framework for Graphene-Based Materials
Peter Wick,* Anna E. Louw-Gaume, Melanie Kucki, Harald F. Krug,
Kostas Kostarelos, Bengt Fadeel, Kenneth A. Dawson, Anna Salvati,
Ester Vzquez, Laura Ballerini, Mauro Tretiach, Fabio Benfenati,
Emmanuel Flahaut, Laury Gauthier, Maurizio Prato, and Alberto Bianco
classification · graphene · nanotechnology ·
structure–activity relationship · toxicology
1. Introduction
Graphene is the enabling material of the 21st century and
there are high expectations for its potential applications. A
clear and consistent system describing the various derivatives
of graphene promotes a precise vocabulary for the family of
graphene-based materials. This will be a prerequisite, for
example, to understand structure–activity relationships in the
context of human health and safety and to avoid general-
izations about the capabilities and limitations of graphene-
based materials. Within the European Unions GRAPHENE
Flagship project, three physical-chemical descriptors specific
for graphene were defined to assist in the classification of
graphene-based materials.
2. Graphene, a Carbon Material with Great
Potential
“Carbon, the basis of all known life on earth, has surprised
us once again.” This statement stems from the press release by
the Royal Swedish Academy of Sciences regarding their
decision to award the 2010 Nobel Prize in Physics “for the
groundbreaking experiments regarding the two-dimensional
material graphene”.
[1]
In just 10 years graphene has become a lodestar for
researchers all over the world. While the media is boosting the
public profile of graphene through the reference to “miracle
material of the 21st century”,
[2,3]
the number of scientific
papers on graphene also exceeded 3000 per year in 2010.
[4]
The commercial interest in graphene is also reflected by
a recent review of the patent landscape of graphene by the
UK Intellectual Property office. Graphene patent applica-
tions have doubled between 2010 and 2012 and that there has
been an order of magnitude difference in the yearly publica-
tion figures over the last five years, with a total of 8416 patents
worldwide by February 2013.
[5]
It is anticipated that graphene
will withstand the normal seven-step sequence for any new
technology: hope–hype–boom–bust–disillusionment–shake-
out–profitability and meet expectations for profitability even
faster than the other carbon allotropes.
[6]
Graphene is
expected to be at the focus of even greater interest for
industrial applications when mass-produced graphene will
have the same outstanding performance as the best samples
produced in research laboratories.
[3]
[*] Dr. P. Wick, Dr. A. E. Louw-Gaume, Dr. M. Kucki, Prof. H. F. Krug
Laboratory for Materials-Biology Interactions Empa
Swiss Federal Laboratories for Materials Science and Technology
Lerchenfeldstrasse 5, CH-9014 St. Gallen (Switzerland)
E-mail: Peter.Wick@empa.ch
Prof. K. Kostarelos
Nanomedicine Laboratory, Faculty of Medical and Human Sciences
National Graphene Institute, University of Manchester (UK)
Prof. B. Fadeel
Nanosafety & Nanomedicine Laboratory—NNL
Division of Molecular Toxicology
Institute of Environmental Medicine
Karolinska Institutet, Stockholm (Sweden)
Prof. K. A. Dawson, Dr. A. Salvati
Center for BioNano Interactions (CBNI), School of Chemistry and
Chemical Biology, University College Dublin (Ireland)
Prof. E. Vzquez
Departamento de Qumica Orgnica
Facultad de Ciencias y Tecnologas Qumias-IRICA
Universidad de Castilla-La Mancha, 13071 Ciudad Real (Spain)
Dr. L. Ballerini, Dr. M. Tretiach
Life Science Department
University of Trieste (Italy)
Dr. F. Benfenati
Department of Neuroscience and Brain Technologies
Istituto Italiano di Tecnologia, Genova (Italy)
Dr. E. Flahaut
CNRS, Institut Carnot CIRIMAT, Universit de Toulouse
Centre Interuniversitaire de Recherche et d’Ingnierie des Matriaux,
Toulouse (France)
Dr. L. Gauthier
CNRS, Universit de Toulouse, Laboratoire Ecologie Fonctionelle &
Environnement - ECOLAB, ENSAT 31326 Castanet Tolosan (France)
Prof. M. Prato
Department of Chemical and Pharmaceutical Sciences
University of Trieste (Italy)
Dr. A. Bianco
CNRS, Institut de Biologie Molculaire et Cellulaire
Laboratoire d’Immunopathologie et Chimie Thrapeutique
Strasbourg (France)
Recently, a roadmap for graphene was published, high-
lighting its future path in the fields of electronics, photonics,
composite materials, energy generation and storage, sensors,
metrology, and biomedicine.
[3]
However, to realize such
potentials, the health and environmental impact of the family
of graphene materials should be thoroughly evaluated.
Although this area will profit from existing knowledge and
concepts for the nanosafety hazard and risk assessment of
other nanomaterials, there are still various hurdles to over-
come.
[7–9]
3. Confusion and Inconsistency in Naming the
Family of Graphene-Based Materials
One concern in graphene re-
search is that, similar to carbon
nanotubes, the term graphene is
used in a generic manner and not
in a precise way by scientists to
describe many graphene-based
materials (GBMs) they have syn-
thesized and studied.
[10]
The in-
consistency in naming arises not
only in connection with the use of
graphene for isolated single-
atom-thick sheets, but also by its
reference to related two-dimen-
sional sheetlike and flake carbon
forms.
[10]
Thus, a clear, consistent,
and widely accepted system of
describing and naming the various
derivatives of graphene still needs
to be developed. Solving this
standardization and nomencla-
ture issue will be of paramount
importance to avoid misleading
understanding and interpretation
amongst all the stakeholders (i.e.,
researchers, industry, govern-
ments, and in particular, regula-
tory authorities), for whom the
science-based assessment of gra-
phene toxicity and environmen-
tal, health, and safety concerns
carries priority.
[11]
4. Classification Approach for Graphene-Based
Materials
Recently, the first nomenclature for 2D carbon forms was
published and the motivation for this article was given by the
statement “Precise names promote precise ideas”.
[10]
Under-
lying this attempt towards a more rational graphene nomen-
clature is a set of definitions based on the fact that graphene
materials should be defined by morphological descriptors. For
example, typical dimensions could be described by graphene-
specific variables such as thickness (layer number) and lateral
size.
[4,10]
In this Essay we would like to build on previous work
that attempted to highlight the importance of graphene
structural characteristics as determinants of their implications
in health and safety. We present an approach (depicted in
Figure 1) that supports the structural notation of GBMs,
a term in line with that proposed previously.
[10]
Our nomenclature model considers the number of gra-
phene layers, the average lateral size, and the carbon-to-
oxygen (C/O) atomic ratio as the three fundamental proper-
ties that cover the largest set of current graphene materials
encountered in practice and that are the subject of biological
testing and of biosafety concern. The two morphological
characteristics are included because GBMs consist of not only
single-layer graphenes but also few-layer graphenes (i.e., 2–10
layers), graphene oxide (GO, normally a single layer),
reduced graphene oxide (rGO; normally a single layer),
graphene nanosheets, ultrafine graphite (i.e., more than 10
Peter Wick heads the Research Laboratory
for Materials–Biology Interactions at Empa,
the Federal Laboratories for Materials Sci-
ence and Technology, Switzerland. He
studied and received his PhD in Cell and
Molecular Biology from the University of
Fribourg (Switzerland). In 2002 he moved
to Empa and started his research career in
nanosafety. He is a board member of the
Swiss Action Plan for Synthetic Nanomate-
rials and member of the editorial board of
the journal Nanotoxicology.
Figure 1. Classification grid for the categorization of different graphene types according to three
fundamental GBM properties: number of graphene layers, average lateral dimension, and atomic
carbon/oxygen ratio. The different materials drawn at the six corners of the box represent the ideal
cases according to the lateral dimensions and the number of layers reported in the literature. The
values of the three axes are related to the GBMs at the nanoscale, but it is feasible to expand the
values to the microscale.
graphene sheets but below 100 nm in thickness), graphene
ribbons, and graphene dots.
[9,10]
Furthermore, it has been
emphasized recently that little is known about the possible
differences in the biological behavior of graphene types
having differnet layer numbers and lateral sizes and these
topics deserve thorough analysis.
[9]
The addition of the C/O ratio as a functional attribute can
be justified by the fact that GBMs are both structurally and
chemically heterogeneous. The family of GBMs includes
materials with widely variable surface oxygen content and
thus, surface chemistry can influence biocompatible disper-
sion potential and colloidal behavior.
[4,9,12]
For example, GO
and rGO are emerging as popular materials in nanocarbon
research, not only as carbon building blocks for biomedical
applications but also as starting materials to produce gra-
phene-based materials.
[10,11]
Our classification framework provides a starting point for
the categorization of distinct graphene types within a grid
arrangement according to three easy-to-measure and quanti-
fiable characteristics. It is also important to state at this stage
that future studies on graphenes biological significance might
reveal other important health and safety assessment criteria
for graphene. Our proposed methodology could be validated
by biologists and nanotoxicologists working with graphene in
order to understand relationships between graphene phys-
icochemical characteristics and safety considerations.
[10,11]
Validation of the proposed methodology will also be given
high priority by authors of this article, as members of the
GRAPHENE Flagship project. The role of the GRAPHENE
Flagship project in the determination of GBM structure–
safety relationships is highlighted below.
GBM Structure–Safety Relationships: The Role of the EU
GRAPHENE Flagship: The GRAPHENE Flagship is a 10-
year project and the European Unions biggest research
initiative ever with a budget of one billion EURO (http://
graphene-flagship.eu/). This project is tasked to take gra-
phene from the realm of academic laboratories into European
society in the space of ten years and to generate economic
growth, new jobs, and new opportunities.
[13]
A specific
research program of the flagship (and in which authors of
this article participate) is intended to reveal the relationshipes
between the material structure and toxicological functions for
different types of graphene materials. The adoption of a clear
nomenclature framework to reflect the different structural
and chemical characteristics of GBMs will greatly help in this
endeavor. In combination with the adoption of benchmarked
biological assays that offer reliably specific toxicological end-
points, we envision a platform by which all graphene materials
will be assessed.
5. Biological and Toxicological Relevance of Funda-
mental Properties of GBMs
The biological significance of GBM properties such as
layer number, lateral dimension, surface chemistry, surface
area, and material purity have already been highlighted.
[4,8,9]
The layer number determines the thickness, the specific
surface area, and bending elasticity with expected outputs
such as higher adsorptive capacity for GBM-type molecules
when the layer number decreases and increased stiffness/
rigidity during cellular interactions when the material thick-
ness increases. As the lateral sizes of GBMs span several
orders of magnitude, from the nanoscale to the microscale
(i.e., 10 nm up to > 20 mm), it is important to specify the
lateral dimension since this parameter determines the max-
imum size and degree of deformability of the material which
are key variables for cellular uptake, renal clearance, trans-
port across the blood–brain barrier, and many other biolog-
ical interactions that depend on particle size.
[4,14,15]
The understanding of the biological relevance of the C/O
atomic ratio requires a more detailed examination. From
a surface science perspective, it should be considered that
members of the GBM family do not have a standard surface.
For example, pristine graphene and GO differ in their surface
hydrophobicity/hydrophilicity. For the former, the surface is
hydrophobic while for GO, surfaces consist of hydrophobic
islands with hydrophilic regions showing various degrees of
basal reactivity. Graphene oxide could be considered as
derivatized graphene with a myriad of oxygen functionalities
due to the introduction of carbonyl, hydroxy, and epoxy
groups on the planar surfaces and edges of the carbon sheets
during graphite oxide exfoliation. Therefore, graphite oxida-
tion endows single- or few-layered GO with the great
advantages of improved solubility or dispersibility in aqueous
solutions and reasonable colloidal stability.
[11,12]
Since the
coverage with oxygen atoms varies depending on the degree
of oxidation during the preparation of GO, it is imperative to
understand that these production processes will contribute to
the inhomogeneity of the final GO product. Typically, GO
with chemical compositions corresponding to a C/O ratio of
4:1 to 2:1 are produced. Graphene can also be transformed
into reduced rGO and this will increase the C/O ratio to
approximately 12:1 but values as large as 246:1 have recently
been reported.
[9,12,16]
The significance of investigating the three fundamental
properties should also be viewed in terms of the relationship
between physicochemical characteristics and the health and
environmental risks of any nanomaterial. The importance of
these characteristics for ecological sustainability is notewor-
thy as these characteristics might modulate GBM–organism
interactions and thus the transfer and impact of GBMs
through the food chain and ecosystems.
[17]
A comparison
study that reviewed toxicological aspects of GO in relation to
its synthesis techniques found that thickness and lateral
dimensions are the structural properties that vary the most.
[7]
In analogy to the requirement that graphene sheets must be of
an appropriate size (i.e., size tuning of the lateral dimension)
to suitably interface with biological systems, evidence is
accumulating that cell viability and toxicity responses can be
modulated by controlling the GBM surface oxygen con-
tent.
[4,18,19]
Although we wish to highlight the biological significance
of these three graphene-specific properties, it is important to
also consider in toxicological assays other generic evaluation
criteria, as is valid for most other nanomaterials. For example,
sample impurities in graphene samples might cause non-
specific interference in toxicological studies
[4]
and, in addi-
tion, once GBMs are introduced into a living system, a new
biological “identity” may be adopted, as determined by the
biomolecules that adsorb to the material surface.
[20]
Graphene
offers a large available surface for the adsorption of proteins
and other biomolecules and this should also be taken into
account when attempting to determine/resolve the overall
health, safety, and toxicity impact of GBMs.
6. Experimental Methodologies to Measure the
Three Classification Parameters
Table 1 summarizes the basic analytical techniques for the
measurement of the three fundamental GBM classification
parameters of interest. Suitable references are also included
which provide a detailed and thorough description of
appropriate experimental methodologies to study these
properties, and it is important that researchers are well-
trained in the appropriate methodologies and incorporate
more than one testing method in their characterization
efforts.
7. Summary and Outlook
Consensus is accumulating that clarity in the nomencla-
ture of GBMs is needed. The proposed classification frame-
work, as an initial reference system for graphene biologists/
toxicologists, will help to determine the role of the three
physicalchemical properties on the health and safety profile
of GBMs. It will contribute to the avoidance of general-
izations about the capabilities and limitations of GBMs that
can raise false expectations and unnecessary safety con-
cerns.
[12]
Other benefits that might stem from the adoption of
the classification and mapping approach are summarized in
Table 2.
There are indeed numerous long-term benefits for all
players in the graphene-biological community if diligence is
shown by all in characterizing their materials and describing
them according to layer number, lateral size, and surface
chemistry rather than using ad hoc sample names.
[4]
Finally, it
is important to keep in mind that the set of critical nanosafety
evaluation criteria for GBMs will grow in the future in order
to reach a consensus on the health and environmental safety
risks of graphene smart materials such as functionalized
GBMs.
The authors gratefully acknowledge financial support from EU
FP7-ICT-2013-FET-F GRAPHENE Flagship project (no.
604391). All authors are members of the EU GRAPHENE
flagship project (Work package 2 Health & Environment). We
thank Andr Niederer (Empa) for assistance with the graphics.
[1] http://www.nobelprize.org/nobel_prizes/physics/laureates/2010/
press.html.
[2] http://news.bbc.co.uk/1/hi/programmes/click_online/9491789.
stm.
[3] K. S. Novoselov, V. I. Falko, L. Colombo, P. R. Gellert, M. G.
Schwab, K. Kim, Nature 2012, 490, 192 – 200.
[4] V. C. Sanchez, A. Jachak, R. H. Hurt, A. B. Kane, Chem. Res.
Toxicol. 2012, 25, 15 – 34.
[5] http://www.ipo.gov.uk/informatics-graphene-2013.pdf.
[6] http://roadtoabundance.wordpress.com/2013/08/14/mass-
production-of-graphene-comes-of-age-graphene-tracker/.
[7] A. M. Jastrze˛bska, P. Kurtycz, A. R. Olszyna, J. Nanopart. Res.
2012, 14, 1320 – 1341.
[8] C. Bussy, H. Ali-Boucetta, K. Kostarelos, Acc. Chem. Res. 2013,
46, 692 – 701.
[9] A. Bianco, Angew. Chem. 2013, 125, 5086 – 5098; Angew. Chem.
Int. Ed. 2013, 52, 4986 – 4997.
[10] A. Bianco, H.-M. Cheng, T. Enoki, Y. Gogotsi, R. H. Hurt, K.
Koratkar, T. Kyotanic, M. Monthioux, C. R. Park, J. M. D.
Tacson, J. Zhang, Carbon 2013, 65,1–6.
[11] D. Bitounis, H. Ali-Boucetta, B. H. Hong, D.-H. Min, K.
Kostarelos, Adv. Mater. 2013, 25, 2258 – 2268.
[12] http://graphene-flagship.eu/?page-id5.
[13] D. R. Dreyer, S. Park, C. W. Bielawski, R. S. Ruoff, Chem. Soc.
Rev. 2010, 39, 228 – 240.
[14] J. Russier, E. Treossi, A. Scarsi, F. Perozzi, H. Dumortier, L.
Ottaviano, M. Meneghetti, V. Palermo, A. Bianco, Nanoscale
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Table 1: Analytical techniques for the classification of GBMs.
[a]
.
GBM-specific property and suitable analytical tools for the classification
of GBMs
Number of layers (thickness) TEM
[21]
AFM
[21]
Raman spectroscopy
[21,22]
Optical absorbance measurements
[23]
Lateral size TEM
[21]
SEM
[21]
AFM
[24]
Atomic C/O ratio XPS
[13,16]
Elemental analysis (ICP-MS)
[25]
[a] TEM: transmission electron microscopy, SEM: scanning electron
microscopy, AFM: stomic force microscopy, ICP-MS: inductively coupled
plasma mass spectrometry.
Table 2: Potential benefits associated with adoption of GBM nomen-
clature.
*
Improved vocabulary/coordinated terminology for structural char-
acterization
*
Classification within the proposed grid prevents ad hoc naming
*
Better comparisons between carbon allotropes
*
Fitting the “molecule of interest” into the classification grid and its
position determines which reference GBM should be included in
biological experiments
*
Standard analytical methodology for benchmarking of materials
*
Future comparative studies possible through minimal material
characterization
*
Aid in structure–activity analogies for predictive toxicology
*
Support the peer-review process
*
Clear-cut guidelines for regulatory purposes
e
![Table 1: Analytical techniques for the classification of GBMs.[a] .](/figures/table-1-analytical-techniques-for-the-classification-of-gbms-9m5tudcj.webp)