Trends in Analytical Chemistry, Vol. 30, No.

3, 2011 Trends

Ecotoxicity of, and remediation

with, engineered inorganic
nanoparticles in the environment
Antoni Sánchez, Sonia Recillas, Xavier Font, Eudald Casals, Edgar González,
Vı́ctor Puntes

This article presents recent developments on the use of inorganic nanoparticles (NPs) for environmental remediation in polluted
soil, water and gas. The number of publications on these topics has grown exponentially in recent years, especially those focused
on wastewater treatment. Among these topics, removal of metals has become the most popular, although some works relate to
the use of nanomaterials for the elimination of nutrients (e.g., nitrogen and some persistent organic pollutants). However, this
growth has not been accompanied by knowledge about the behavior of NPs once used and released into the environment. In this
article, we also comment upon the current situation with respect to NP toxicology (nanotoxicology). A remarkable number of
different toxicology tests has been applied to NPs, often making it very difficult to interpret or to generalize the results. We
analyze in detail the bioluminescence, Daphnia magna and other tests, and give some preliminary results obtained in our work.
ª 2011 Published by Elsevier Ltd.

Keywords: Bioluminescence test; Daphnia magna; Environmental remediation; Metal removal; Nanoparticle; Nanotoxicology; Nanowaste;
Pollution; Toxicity; Wastewater treatment

1. Introduction economy will be $1 trillion and that 2

Antoni Sánchez, Sonia Recillas,
Xavier Font
Department of Chemical
Engineering, Escola Tècnica
Superior dÕEnginyeria,
Universitat Autònoma de
Barcelona, 08193 Bellaterra,
Spain
Eudald Casals, Edgar González,
Vı́ctor Puntes
Institut Català de
Nanotecnologia, Campus de la
Universitat Autònoma de
Barcelona, 08193 Bellaterra,
Spain
Vı́ctor Puntes*
Institut Català de Recerca i
Estudis Avançats, Passeig Lluı́s
Companys, 23, 08010
Barcelona, Spain
*
Corresponding author.
Tel.: +34 935 868 013;
Fax: +34 935 868 020;
E-mail: victor.puntes@uab.es
As a result of advances in methods of
producing nanoparticles (NPs) with con-
trolled composition and properties, as well
as the incipient development of protocols
for large-scale synthesis, in recent years
use and production of engineered inor-
ganic NPs has increased exponentially.
These nanostructures are the base for
manufacturing new materials with a
variety of applications:
 health sector and life sciences (18%);
 chemicals (12%);
 energy, communication and informa-
tion technologies, transportation, and
environmental (around 8–9% each);
and,
 construction, household products, de-
fense and security, aerospace industry,
personal care, food industry and textiles
(1–6%) [1].
Some of these nanostructures have been
inserted as raw matter in the productive
sector and support the research tasks. The
National Science Foundation estimates
that by 2015 the impact on the global
million workers will be employed in
nanotechnology [2].
NPs have also found applications di-
rectly in environmental remediation [3–
5], as reflected in the increasing number of
publications and the amount of funding
for remediation projects. For instance,
nearly $1 billion were allocated to the US
Environmental Protection Agency (EPA)
in 2009 for environmental remediation
[6].
The use of NPs for environmental
remediation could help to reduce this cost
but the benefit obtained has to be balanced
with the potential risks. In this context, it
is necessary to assess the environmental
risk factors of exposure to engineered NPs.
Apart from exposure through intended
use, unwanted dispersion (spill) or
(nano)waste management also have to be
considered as critical pathways to intro-
duce NPs in the environment.
The response of the NPs in the envi-
ronment can be extremely complex and
diverse, depending on a variety of para-
meters involved, which makes it difficult

0165-9936/$ - see front matter ª 2011 Published by Elsevier Ltd. doi:10.1016/j.trac.2010.11.011 507
Trends
to know their environmental fate precisely. They may be
aggregated into microscopic particles or embedded in
exposed materials; they may corrode and dissolve, or
suffer morphological modifications. The surfaces of the
NPs experience constant modifications as they evolve
through the environment, and it is through their sur-
faces that they interact with their environment. Thus, in
biological environments, the toxic effects of NPs will
depend on not only the initial morphological properties,
composition, size, additives and synthesis method em-
ployed, but also the physico-chemical evolution in the
surroundings [7–9].
Auffan et al. [10] pointed out that the chemical sta-
bility at physiological redox conditions appears to be a
condition for non-toxicity in metallic NPs. Nevertheless,
metallic NPs with strong oxidant or reductive properties
can be cytotoxic and genotoxic [10], so it is necessary to
evaluate risk factors by considering morphology, size,
method of producing NPs, type of target and conditions
of interaction [11,12].
Recently, a model to predict NP toxicity was proposed
with the available electronic energy levels in the NP
structure matched up with the oxidation potentials of
reactions that would remove antioxidants from cells or
generate reactive oxygen species (ROSs) {e.g., hydrogen
peroxide or superoxide ions (O2-) [13]}. Where these two
values overlap, it means that the NPs can accept elec-
trons in these processes and cause oxidative stress in
cells –by eliminating antioxidants or producing ROSs.
The objective of this article is to present, on the one
hand, the most recent developments on the use of NPs
for environmental remediation and, on the other hand,
the first attempts to assess NP toxicology and the tests
available in the scientific literature.
2. Environmental remediation applications of
inorganic nanoparticles
While industrial sectors involving semiconductor,
memory and storage technologies, display, optical and
photonic technologies, energy, biomedical and health
sectors produce the most products containing nanoma-
terials, nanotechnology is also used as an environmental
technology to protect the environment through pollu-
tion prevention, treatment and clean-up. Nanoremedia-
tion has the potential not only to reduce the overall costs
of cleaning up large-scale contaminated sites, but it can
also reduce clean-up time, eliminate the need for treat-
ment and disposal of contaminated soil, and reduce some
contaminant concentrations to near zero – all in situ [6].
In general, NPs present some natural attributes to be
applied to environmental remediation. Due to its reduced
size, the NP surface grows exponentially for the same
density as the diameter shrinks. Due also to their small
size, the mobility of NPs in solution is high and the whole
508 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 30, No. 3, 2011
volume can be quickly scanned with small amounts of
NPs. For example, at a rough estimate, a 10-nm gold NP
in water at room temperature will experience Brownian
relaxation of the order of 1 ns, and each Brownian step
in solution will move it 10–20 nm. A typical NP con-
centration of few nM will explore the total volume in the
order of tens of milliseconds [14]. In addition, due to its
reduced size and their large radii of curvature, the NP
has a surface that is especially reactive (mainly due to
high density of low coordinated atoms at the surface,
edges and vortex).
These unique properties can be employed to degrade
and scavenge pollutants (Fig. 1). The species absorbed
onto the NP can be removed by applying mild (and
affordable) gravitational (centrifugal) or magnetic (in the
case of magnetic NPs) gradients (Fig. 2). The ability of
the inorganic NPs to trap pollutants is well known for
cations (e.g., As, Cr, Ni, and Cd) [15]. In the case of
organic molecules, NPs can be functionalized specifically
to trap particular molecules. Finally, the use of the NPs
as photocatalysts to promote the complete degradation of
organic matter is also being explored.
2.1. Removal of metals from streams
Removal of metals with NPs is a most abundant field in
the literature. Typically, the papers describe an engi-
neered NP to remove a specific metal, which is present at
low amounts in drinking water.
The paradigm is use of magnetic iron-oxide NPs to
remove As ions from drinking water. The As ions
spontaneously absorbs onto the iron-oxide NPs and the
complex is removed using weak magnetic fields [15].
Lisha et al. [16] reported another example, in which
gold NPs supported on alumina were used as adsorbents
in order to remove inorganic mercury from drinking
water.
Zero-valence iron NPs found application for the effi-
cient removal of different metals {e.g., Cr (VI) [17], U(IV)
and U(VI) [18] and Co(II) [19]}.
Also, we have explored the ability of cerium-oxide NPs
for the removal of Cr(VI) through adsorption of the metal
onto the surface of the NPs [20].
Other strategies involve the functionalization of NPs,
as in the work of Skubal et al. [21], where the surfaces of
titanium-dioxide NPs were modified with bidental che-
lating agent thiolactic acid (TLA) to remove aqueous
cadmium from simulated wastewaters.
The drawbacks in these papers are typically the use of
synthetic water and the lack of knowledge about the
detailed mechanisms of interaction between NPs and
metals. When using synthetic water, it is clear that the
results may not be translated into the real world, and
some problems (e.g., scale up, cost evaluation, and
downstream recuperation of NPs) are often omitted.
Scientific knowledge is most developed in the case of the
interactions of NPs and metals, and typically adsorption
Trends in Analytical Chemistry, Vol. 30, No. 3, 2011 Trends

Figure 1. Schema of different methods of clean up using nanoparticles (NPs). 1) NPs that degrade contaminants in situ (e.g., photocatalysis of
organic matter using TiO2 NPs). 2) NPs that adsorb contaminants (e.g., CaCO3 adsorbs Ni, and Fe3O4 adsorbs As and Cr). Adsorbed metals lose
their toxicity. Moreover, NPs can be separated from media using magnetic or gravitational fields. 3) NPs conjugated to molecules that adsorb
contaminants (e.g., NPs conjugated to cyclodextrins that adsorb persistent organic pollutants). Moreover, NPs can be separated from media using
magnetic or gravitational fields.

Figure 2. Magnetic removal of contaminated water using Fe3O4 nanoparticles (NPs). Left: Fe3O4 NPs mixed with sludge in 1:1 proportion. Right:
NPs trap pollutants and, due to the magnetic character of the NPs, the composite is separated from liquid driven by a magnetic field. Image was
taken 1 min after the magnet was placed.

is the main mechanism {e.g., Langmuir and Freundlich
isotherms have been proposed for arsenic removal [22]},
and, in some cases, organic modifications of NPs have
been synthesized to enhance the efficiency of metal re-
moval [23].
Table 1 summarizes some of the main results recently
published in this field. Although interesting, we can
observe that some of them lacked the specific charac-
teristics of proposed model mechanisms, and most of
them were conducted with synthetic water.
2.2. Removal of nutrients from streams
Phosphorous and nitrogen in its different chemical forms
are normally considered nutrients. There are many
factors that influence nutrient behavior in the environ-
ment and their interrelationships may be complex.
Nitrogen and phosphorus are the main plant nutrients
that warrant concern with respect to surface-water
systems. NPs may absorb them or transform their oxi-
dation state into that of environment-friendly species.
Some works have been published on the elimination of
nitrogen with NPs, although literature is scarce on this
topic {e.g., Choe et al. [24] reported the reductive deni-
trification by nanoscale zero-valence iron}. Apart from
this simple method of reduction of nitrate with iron,
recent studies have been focused on more complex
mechanisms for nitrate removal {e.g., the photocatalytic
reduction reaction with pure TiO2 NPs [25] or TiO2 NPs
doped with Bi3+ [26]}. The use of iron is appealing, but
the pyrophoricity of the iron NPs makes it very difficult
to preserve them in the zero-valence state before their
real application in the environment.

http://www.elsevier.com/locate/trac 509
Trends Trends in Analytical Chemistry, Vol. 30, No. 3, 2011

Table 1. Some of the results obtained with nanoparticles to remove metals from water

Removed metal(s) Nanoparticle Mechanism Water Ref.

As(III), As(V) Iron(III)–titanium(IV) binarymixed oxide Adsorption Synthetic and real [22]
Mo(VI) Maghemite Adsorption Synthetic [70]
Cr(VI) Chitosan-Fe0 Reduction Synthetic [71]
Pb(II) Titanium phosphate Adsorption Synthetic [72]
Pb(II) Silica-alumina Adsorption Synthetic [73]
Cr(VI) Zero-valence iron Reduction Synthetic [17]
Cu(II) Fe3O4 chitosan bound Adsorption Synthetic [74]
U(IV), U(VI) Zero-valence iron Reduction Real [18]
Hg(II) FeS Adsorption Synthetic [32]
Cd(II) TiO2 Adsorption/Reduction Synthetic [21]
Co(II) Zero-valence iron Adsorption Synthetic [19]
Pb(II), Ni(II) Fe3O4 chitosan bound Adsorption Synthetic [75]
Au, Ag, Pt, Pd Several Adsorption Real [76]
Fe(III) Fe3O4 chitosan bound Adsorption Synthetic [77]

Only Martin et al. [27] have reported the use of NPs to
remove and to recover phosphate, in this case, from
municipal wastewaters using ferric-oxide NPs. In the
same way, our research group has promising results
with cerium-oxide NPs and phosphate in synthetic
water. These results regarding phosphate elimination are
very encouraging since, nowadays, phosphate elimina-
tion must be performed chemically or through a complex
biological process.
2.3. Removal of organic pollutants from streams
Research on the removal of organic compounds with
several types of NP has been initiated in several studies
(Table 2). Special attention is being focused on persistent
organic pollutants (e.g., DDT). Typically, the mecha-
nisms are not well understood but the interest and the
difficulty of eliminating traces of these compounds are so
great that the first publications deserve attention.
Ghauch et al. [28] reported interesting results on the
elimination of antibiotics from water using iron NPs, and
Wang et al. [29] presented bismuth-doped TiO2 NPs for
photocatalytic degradation of rhodamine B, a commonly
used dye, among others.
In general, results obtained in the degradation of or-
ganic pollutants with NPs are very promising and rep-
resent a significant step forward in the removal of
persistent organic pollutants. Table 2 summarizes some
of the main results recently published in this field. As can
be observed, photocatalysis and/or the use of other
reactants (e.g., H2O2 and H2) are common mechanisms
used to degrade the pollutants. However, the
combination of a biological process with NPs to degrade

Table 2. Some of the results obtained with nanoparticles to remove organic pollutants from water

Organic pollutant Nanoparticle Comment Ref.

Trichloroethene and chlorobenzene Palladium/magnetite Photocatalytic degradation [78]
Trichloroethylene Powder-activated carbon and Nanoparticles enhance the adsorption on [79]
Fe2O3,TiO2, and SiO2 powder-activated carbon
Rhodamine B Fe3O4 Chemical reaction with H2O2 as oxidant [80]
Acetone, benzene, and toluene In(OH)3 Photocatalytic degradation (UV) [81]
Rhodamine 13 BiFeO3 Fenton reaction [82]
Trichloroethylene Bimetallic particles of nickel on iron, Catalytic breaking of C–Cl bond [83]
supported on functionalized carbon
nanotubes
Alachlor and phenanthrene Copper oxide Chemical reaction with H2O2 as oxidant [84]
Trichloroethylene Bio-Pd Dechlorination with bio-Pd, hydrogen gas [30]
and formic acid
Acid Black 24 Fe0 Photocatalytic degradation (UV) and H2O2 as [85]
oxidant
Hexachlorocyclohexanes Fe0 Reaction rather than sorption was the [86]
operative mechanism for pollutant removal
Acid Black-24 TiO2 and Fe0 Photocatalytic degradation (UV) [87]
Anthracene-9-carbonxylic acid CdSe Photocatalytic degradation (green [88]
monochromatic light)
Lindane Fe0 Catalytic degradation [89]

510 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 30, No. 3, 2011
trichloroethylene [30] is remarkable, as it represents a
new approach to the biological treatment of wastewaters.
2.4. Polluted soil and gases
2.4.1. Nanoparticles for soil remediation. This is a topic
with less research compared to removal of pollutants
from water or wastewater. In this situation, the advan-
tages of the Brownian dispersion are lost. Typically, the
physical phenomena involved in soil-NP interactions are
not understood, so the results obtained in this field are
only preliminary. Moreover, NPs are not in soil but they
are spiked into it – far from the real case, as the intro-
duction of NPs in soil is one of the most difficult aspects
to consider in in situ soil bioremediation. For ex situ
remediation, soil columns are typically set up, and a
liquid suspension of NPs is added to extract or to
immobilize the contaminants (typically metals or per-
sistent organic compounds). Accordingly, some inter-
esting papers have been published. Xu and Zhao [17]
reported the reductive immobilization of chromate in
water and soil using stabilized iron NPs in laboratory
batch and column experiments, whereas other studies
reported the immobilization of some organic pollutants
in soils and sediments [31,32].
2.4.2. Gas treatment with nanoparticles. This topic can
be considered the most emerging field in the use of NPs.
Although the investigation of nanomaterials specifically
designed to clean polluted gases has just started and the
results must often be seen as preliminary, some studies
have recently been published on gas treatment using
NPs, apart from the large number of published studies on
nanomaterials used as a sensors for gas detection and
measurement [33], which comprises another research
area.
Toxic gases {e.g., carbon monoxide [34], toluene [35]
and gases containing chlorinated compounds [36]} have
been successfully removed with platinum, titanium-
oxide and Fe-Pd bimetallic NPs, respectively. Although it
is probable that new applications will appear in the years
ahead, the potential of using titanium-oxide NPs for the
reduction of polluted gases seems to be a accepted [37].
TiO2 NPs photocatalyze the conversion of airborne pol-
lutants (principally, volatile organic compounds and
nitrogen oxide) into less harmful compounds. Thus,
there have been some attempts to maintain the cleanli-
ness of buildings in areas with dense traffic by applying
TiO2-NP paints [38].
3. Standardized toxicity tests for inorganic
nanoparticles
The question here is whether the potential benefits
of using NPs present additional risks. How NPs
may interact with toxic ionic and molecular species –
Trends
capturing and destroying them without presenting any
hazard to microorganisms – may be the key, since
microorganisms have defense mechanisms that molec-
ular species do not have. Otherwise, if the cleaning
product is itself toxic, interest in NPs for environmental
remediation will be very limited.
Several studies of the toxicity of uncoated, water-sol-
uble, colloidal fullerenes (C60) have been done. In one
study, the 48-h LC50 (median lethal concentration) in
Daphnia magna was determined to be about 800 ppb
[39]. In another, using largemouth bass (Micropterus
salmoides), lipid peroxidation in the brain and glutathi-
one depletion in the gill were observed after exposure to
0.5 ppm of C60 for 48 h [40].
The toxicity of some metal-oxide NPs has also been
studied. TiO2 absorbs substantial UV radiation, yielding,
in aqueous media, hydroxyl species that may cause
substantial damage to DNA [41,42]. Other studies found
that Al2O3 NPs could reduce root growth due to per-
turbation of the microbial composition of the soil [43],
raising concerns since many food chains depend on the
benthic and soil flora and fauna, which could be affected
by such NPs. However, Yang and Watts [43] observed
that alumina NPs do not induce any detectable effects
on seed-root growth using root-elongation tests on
C. sativus, although Zn and ZnO NPs significantly
inhibited seed germination and root growth [44]. In this
area, Warheit et al. proposed a set of basic toxicity tests
to manage TiO2 risk [45]. Since nanotoxicology is still at
its early stages in terms of available data, any scientific
contribution on environmental risks of NPs should help
to regulate the use and the production of nanoengi-
neered materials.
3.1. Wastewater-based tests
Although not especially standardized for NP suspen-
sions, several tests commonly applied to wastewater, or
even drinking water, have been used to determine the
toxicity of NPs. These tests are easy to carry out in the
laboratory, but often interpretation of the results with
NPs is not straightforward, since the ultimate cause of
toxicity is rarely known. Some of these tests are detailed
below.
3.1.1. Bioluminescence test. A standardized test, called
Microtox, has been used with NPs. This method is based
on the percentage of decrease in the amount of light
emitted by bioluminescent marine bacterium Vibrio
fischeri upon contact with a filtered sample at pH 7
containing a known concentration of NPs [46]. The
effective concentration, EC50, is defined as the concen-
tration that produces 50% reduction of light. EC50 can be
measured after 5-min and 15-min contact time. Biolu-
minescent tests must be performed under a sodium-
chloride concentration of 22%, according to the
manufacturerÕs instructions.
http://www.elsevier.com/locate/trac 511
Trends
Our previous results [47] did not show toxicity using
this test with iron oxide, silver and gold NPs, but other
works have presented different results. For example, in
the case of titanium-oxide NPs, low levels of toxicity
were found with the bioluminescence test and EC50
could not be measured [48], whereas boron NPs showed
high levels of toxicity and a value of EC50 in the range
56–66 mg/L, depending upon the age of the solution.
Differences could be attributed to different NP prepara-
tions. For example, we should remember that, in every
NP synthesis, some ions are remnants. A paradigm is
provided by commercial samples of colloidal silver in-
tended for water purification, where the amount of ionic
silver may be as high as the 90% with respect the total
silver content. This factor is mostly neglected and may
have a large effect on the results [49–51].
Figure 3. (Top) Transmission electron microscopy images of model nanoparticles (NPs) produced in our laboratory and used for the experiments
mentioned in the text. Scale bars are 20 nm. (Bottom) Image of anaerobic bacteria after a toxicology test with silver NPs. Scale bar is 200 nm.
512 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 30, No. 3, 2011
3.1.2. Daphnia magna. This is probably the most
extensive test to evaluate NPs toxicity [52]. D. magna is a
cladoceran freshwater water flea native to northern and
western North America. Domesticated, it is widely used
as a laboratory animal for testing ecotoxicity. In this test,
acute 48-h toxicity assays with D. magna are conducted
following Organization for Economic Co-operation and
Development (OECD) protocols [53]. Groups of 10 neo-
nates (<24-h old) are exposed to 20 mL of test concen-
trations in 50 mL flasks with no food present. At the end
of exposure, the number of immobile animals is recorded
and median lethal concentration levels (LC50) are esti-
mated.
D. magna is often very sensitive to the presence of NPs,
and there is a large number of publications on this topic,
some of which distinguish between death and
Trends in Analytical Chemistry, Vol. 30, No. 3, 2011
immobilization of D. magna [54]. In their review, Farré
et al. [52] reported the main data observed when expos-
ing D. magna to several types of NPs. In this and other
works [54], ZnO and TiO2 and C60 fullerenes appeared to
be very toxic to D. magna, whereas Fe3O4 NPs exhibited a
low effect. However, our experience is that the results of
this test are somewhat erratic and a lot of replications
must be carried out to obtain a reliable value of LC50.
3.1.3. Other aquatic organisms. Although not so exten-
sively used, other superior organisms have been used for
testing NP toxicity, the most typical being Danio rerio
(zebrafish) [52] and Oncorhynchus mykiss (rainbow trout)
[55]. Nevertheless, we should mention that, in these
tests, overall toxicity can be assessed, but the objective is
often to find damage to specific organs after exposure to
NPs or accumulation of NPs.
Acute toxicity tests are significant to determine the
effect of a sudden load of NPs dumped into a wastewater-
treatment plant or a natural water stream. However, it is
also important to investigate the effect of chronic tests
and bioaccumulation of NPs to establish the effect of
continuous spread of NPs in the environment. Bioaccu-
mulation assays have been done with D. magna, finding
that effects of TiO2 toxicity are much greater at 21 days
than at 72 h and greater at 72 h than at 48 h [56], or
investigating the effect of the size of TiO2 NPs on chronic
toxicity [57].
Mice and marine polychaete have also been used to
test bioaccumulation of gold and TiO2 NPs, respectively
[58,59], but showing low rates of accumulation of NPs
in the body.
Finally, toxic effects of NPs on algae are of the utmost
importance since this group provides the food base for
most marine food chains. Interestingly, in this case,
many authors stress the importance of NPs degrading
in aquatic environments (i.e. the role of the ions re-
leased from the NPs). Navarro et al. [60] exposed
Chlamydomonas reinhardtii to silver NPs and silver ions
(AgNO3) and obtained evidence that pointed to both
ions liberated from the NPs and the NPs themselves
being sources of toxicity. A similar study of freshwater
microalga Pseudokirchneriella subcapitata exposed to ZnO
and ZnCl2 found similar results in the case of ZnO NPs
[61].
3.2. Germination, earthworms and other tests
These tests have been used to evaluate the toxicity of
different NPs – to a minor extent germination [43,47]
and earthworms [62]. Too few data are available to
extract reliable conclusions about the validity of these
tests except for the specific situation where they have
been carried out.
It is evident that practically any microorganism in
which a feasible method to determine its biological
activity exists is suitable to determine toxicity when
Trends
exposed to NPs. This is the case of liquid respirometry
for aerobic microorganisms, in which oxygen uptake
rate (OUR) is compared in the presence and in the
absence of NPs, and, the case of anaerobic populations,
in which the specific biogas/methane production is
compared. Several studies have been published on this
topic. For example, Nyberg et al. [63] and Barrena et al.
[47] tested the effect of several NPs on the production of
biogas by selected anaerobic consortia. The results
showed that, at the concentrations assayed, NPs were
not toxic in most cases, although the study focused only
on some specific types of NP. Interestingly, an accu-
mulation of NPs in or on microorganisms was detected
by transmission electron microscopy (Fig. 3). Also,
several reviews have been published on the toxicity of
NPs in aerobic environments [11]. Some of these studies
were alert on the fact that the in vitro toxicity value
obtained with a selected specie could not be applied to
other microorganisms [64] because some important
factors are often forgotten {e.g., NP size, the specific
surface characteristics of the NPs [43] and their state of
agglomeration [65]}.
4. Toxicology data
Clearly, it would be necessary to have a database com-
bining all organisms and NPs and the toxicity effects for
each species, but the amount of work necessary makes
the costs almost impossible to face [66]. In their inter-
esting article, Choi et al. [66] commented that it is very
important to gather information about the toxicity of
NPs in order to have proper regulation, but they esti-
mated for the USA that costs for testing existing NPs
ranges from $249 m for optimistic assumptions about
NP hazards. Also, the time taken to complete testing is
likely to be very high (34–53 years), if all existing
nanomaterials are to be thoroughly tested. Of course,
maturation of the field, understanding of the mecha-
nisms involved and common sense will significantly
shorten the time, and reduce the cost and the effort to
regulate nanomaterials properly.
Table 3 shows a first attempt to present toxicological
effects for inorganic NPs (acute toxicity). This table is
based in our previously published data [47] and our
unpublished data. Although incomplete, it can serve as a
basis to develop new studies on toxicity of NPs, including
new methodologies and types of NP. As observed in
Table 3, some trends can be deduced {e.g., the high
toxicity of cerium-oxide NPs, the innocuous character of
gold NPs, and the surprisingly low toxicity of silver NPs,
although other authors have found different results with
nitrifying bacteria [67]}.
Finally, it is important to report data on the charac-
terization of the NPs used in toxicity tests. Size,
methodology of synthesis, specific surface, solvent and
http://www.elsevier.com/locate/trac 513
Trends Trends in Analytical Chemistry, Vol. 30, No. 3, 2011

Table 3. Acute toxicological data on inorganic nanoparticles (NP). Concentration of NPs was the maximum reached in the laboratory without
agglomeration. Based on Barrena et al. [47], Garcia et al. [90], and other unpublished studies of the authors. Since NP concentration was dif-
ferent in each toxicity test, the file concentration range corresponds to the range of concentrations used in the different tests. Bioluminescence,
Daphnia magna, aerobic and nitrification consortia test results are expressed as EC50 values. The Germination test values are expressed as Ger-
mination Index. Anaerobic toxicity test is expressed as percentage of biogas reduction

Nanoparticle
CeO2 TiO2 Fe3O4 Au Ag
£ ¥ ¥
Solvent HMT TMAOH TMAOH Trisodium citrate Sodium borohydrate
Concentration range (lg/mL) 0.02–0.57 0.48–1.01 18–116 oct–62 16–100
Mean size (nm) 6.5 7.5 6 10 29
Bioluminescence test (Microtox) 0.021 mg/mL EC50 >45% of the EC50 >45% of the EC50 >45% of the EC50 >45% of the
tested concentration tested tested tested concentration
concentration concentration
Daphnia magna (Standard test) 0.012 mg/mL 0.016 mg/mL 2.3 · 10 4 mg/mL No data No data
Germination test (several seeds 0% 40–60% 70–75% 100–120% 75–95%
tested)
Anaerobic consortium (biogas 87% No effect No effect 18% No effect
production)
Aerobic consortium (oxygen 0.18 mg/mL No effect No effect No effect 33% inhibition at
uptake rate) 0.13 mg Ag-NP/mL
Nitrification consortium (oxygen 0.21 mg/mL No data No effect No effect No effect
uptake rate)
¥
Tetramethylammonium hydroxide.
£
Hexamethyl tetramine.

formulation used are factors that definitively affect the 6. Conclusions

toxicity of NPs [47,67].
5. Nanoparticles as a waste
Another important issue to consider when dealing with
NPs is what to do when they are to be treated or disposed
of as a waste. At present, it is too early to have abundant
literature on this topic, but some reviews have been
published. For example, Bystrzejewska-Piotrowska et al.
[68] pointed out that any approach proposed for the
treatment of nanowaste requires understanding of all the
properties of the nanowaste (i.e. chemical, but also
physical and biological).
Among the techniques to recycle used NPs, noble-
metal NPs will be the object of recovery because of their
high price. For other NPs, bioaccumulation using some
species of plants and fungi can be a way to remove NPs
for water, air and soil, although the mechanisms of these
processes are not currently known. Finally, we should
not underestimate the effect in the long run of handling
waste-containing nanomaterials. In our laboratory, we
have chosen to destroy the NPs by irreversible aggre-
gation or dissolution, and then to process the resulting
material conventionally.
More recently, Brar et al. [69] focused on the presence
of engineered NPs in wastewater and wastewater sludge,
and their role when being landfilled or applied to soil.
They concluded that there were too many unanswered
questions regarding the fate and the impact of NPs to
make any prediction about them.
This article summarizes the current situation of NPs
and their application with respect to environmental
remediation and toxicity effects. In conclusion, we can
state that the large number of publications involving
NPs for remediation of selected environments (espe-
cially wastewater) is not accompanied by a deep
knowledge about the long-term, and sometimes even
the short-term, effects on the environment of handling
NPs. Greater efforts in research are necessary to help
stakeholders promote science-based regulations for
nanotechnology, and for nanotechnology to be on
good terms with society.
Acknowledgements
Financial support was provided by the Spanish Ministerio
de Medio Ambiente y Medio Rural y Marino (Project Exp.
007/RN08/03.1).
References
[1] Research and Markets, Government Policy and Initiatives in
Nanotechnology Worldwide 2007 (http://www.researchandmarkets.
com/reports/556274/).
[2] National Science Foundation, Press Release 08-155 <http://
www.nsf.gov/news/news_summ.jsp?cntn_id=112234>, 2010
(accessed 16.11.2010).
[3] S. Joo, I. Cheng, Nanotechnology for Environmental Remediation,
Springer, New York, USA, 2006.
[4] K.J. Klabunde, L. Erickson, O. Koper, R. Richards, Review of
Nanoscale Materials in Chemistry: Environmental Applications,

514 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 30, No. 3, 2011 Trends

ACS Symp. Ser. 1045, American Chemical Society, Washington [38] Building And Nanotech <http://www.nanowiki.info>, 2010
DC, USA, 2010. (accessed 16.11.2010).
[5] S.N. Singh, R.D. Tripathi, Environmental Bioremediation Tech- [39] E. Oberdorster, Abstr. Papers Am. Chem. Soc. 227 (2004)
nologies, Springer, Berlin, Germany, 2007. U1233.
[6] B. Karn, T. Kuiken, M. Otto, Environ. Health Perspect. 117 (2009) [40] E. Oberdorster, Environ. Health Perspect. 112 (2004) 1058.
1823. [41] R. Dunford, A. Salinaro, L.Z. Cai, N. Serpone, S. Horikoshi, H.
[7] T.J. Brunner, P. Wick, P. Manser, P. Spohn, R.N. Grass, L.K. Hidaka, J. Knowland, FEBS Lett. 418 (1997) 87.
Limbach, A. Bruinink, W.J. Stark, Environ. Sci. Technol. 40 [42] H. Hidaka, S. Horikoshi, N. Serpone, J. Knowland, J. Photochem.
(2006) 4374. Photobiol., A 111 (1997) 205.
[8] J.Y. Choi, S.H. Lee, H. Bin Na, K. An, T. Hyeon, T.S. Seo, [43] L. Yang, D.J. Watts, Toxicol. Lett. 158 (2005) 122.
Bioprocess Biosyst. Eng. 33 (2010) 21. [44] D.H. Lin, B.S. Xing, Environ. Pollut. 150 (2007) 243.
[9] X.K. Hu, S. Cook, P. Wang, H.M. Hwang, Sci. Total Environ. 407 [45] D.B. Warheit, R.A. Hoke, C. Finlay, E.M. Donner, K.L. Reed, C.M.
(2009) 3070. Sayes, Toxicol. Lett. 171 (2007) 99.
[10] M. Auffan, J. Rose, M.R. Wiesner, J.Y. Bottero, Environ. Pollut. [46] Deutches Institut für Normung (DIN), DIN 38412, Part 34, DIN,
157 (2009) 1127. Berlin, Germany, 1991.
[11] B. Nowack, T.D. Bucheli, Environ. Pollut. 150 (2007) 5. [47] R. Barrena, E. Casals, J. Colon, X. Font, A. Sanchez, V. Puntes,
[12] M.R. Wiesner, G.V. Lowry, P. Alvarez, D. Dionysiou, P. Biswas, Chemosphere 75 (2009) 850.
Environ. Sci. Technol. 40 (2006) 4336. [48] N. Strigul, L. Vaccari, C. Galdun, M. Wazne, X. Liu, C. Christod-
[13] E. Burello, A.P. Worth, Nanotoxicology (2010) (doi:10.3109/ oulatos, K. Jasinkiewicz, Desalination 248 (2009) 771.
17435390.2010.502980). [49] T. Pfaller, R. Colognato, I. Nelissen, F. Favilli, E. Casals, D. Ooms,
[14] 1012 NPs in one mL. Each NP is responsible to scan a volume H. Leppens, J. Ponti, R. Stritzinger, V. Puntes, D. Boraschi, A.
which is 10 12 mL, which in cubic nm is 109 nm3. The volume of Duschl, G.J. Oostingh, Nanotoxicology 4 (2010) 52.
one 10-nm Au NP is: 523 nm3, so each NP has to take 109/523 [50] P. Rivera-Gil, G. Oberdorster, A. Elder, V. Puntes, W.J. Parak, ACS
steps to visit its whole corresponding volume – about 106 steps; Nano 4 (2010) 5527.
with 10% efficiency, that makes a need for 107 steps, and, with [51] D.B. Warheit. Nano Lett. 10 (2010) 477.
each taking 1 ns, that means that the whole volume is explored [52] M. Farré, S. Pérez, L. Kantiani, D. Barceló, Trends Anal. Chem. 27
every 10 2 s. (2008) 991.
[15] C.T. Yavuz, J.T. Mayo, W.W. Yu, A. Prakash, J.C. Falkner, S. [53] C. Barata, D.J. Baird, Aquatic Toxicol. 48 (2000) 195.
Yean, L.L. Cong, H.J. Shipley, A. Kan, M. Tomson, D. Natelson, [54] X.S. Zhu, L. Zhu, Y.S. Chen, S.Y. Tian, J. Nanopart. Res. 11
V.L. Colvin, Science (Washington, DC) 314 (2006) 964. (2009) 67.
[16] K.P. Lisha, Anshup, T. Pradeep, Gold Bull 42 (2009) 144. [55] G. Federici, B.J. Shaw, R.D. Handy, Aquatic Toxicol. 84 (2007)
[17] Y.H. Xu, D.Y. Zhao, Water Res. 41 (2007) 2101. 415.
[18] M. Dickinson, T.B. Scott, J. Hazard. Mater. 178 (2010) 171. [56] X.S. Zhu, Y. Chang, Y.S. Chen, Chemosphere 78 (2010) V.
[19] C. Uzum, T. Shahwan, A.E. Eroglu, I. Lieberwirth, T.B. Scott, K.R. [57] K.T. Kim, S.J. Klaine, J. Cho, S.H. Kim, S.D. Kim, Sci. Total
Hallam, Chem. Eng. J. 144 (2008) 213. Environ. 408 (2010) 2268.
[20] S. Recillas, J. Colón, E. Casals, E. González, V. Puntes, A. Sánchez, [58] T. Galloway, C. Lewis, I. Dolciotti, B.D. Johnston, J. Moger, F.
X. Font, J. Hazard. Mater. 184 (2010) 425. Regoli, Environ. Pollut. 158 (2010) 1748.
[21] L.R. Skubal, N.K. Meshkov, T. Rajh, M. Thurnauer, J. Photochem. [59] C. Lasagna-Reeves, D. Gonzalez-Romero, M.A. Barria, I. Olmedo,
Photobiol., A 148 (2002) 393. A. Clos, V.M.S. Ramanujam, A. Urayama, L. Vergara, M.J. Kogan,
[22] K. Gupta, U.C. Ghosh, J. Hazard. Mater. 161 (2009) 884. C. Soto, Biochem. Biophys. Res. Commun. 393 (2010) 649.
[23] S.H. Huang, D.H. Chen, J. Hazard. Mater. 163 (2009) 174. [60] E. Navarro, F. Piccapietra, B. Wagner, F. Marconi, R. Kaegi, N.
[24] S. Choe, Y.Y. Chang, K.Y. Hwang, J. Khim, Chemosphere 41 Odzak, L. Sigg, R. Behra, Environ. Sci. Technol. 42 (2008) 8959.
(2000) 1307. [61] N.M. Franklin, N.J. Rogers, S.C. Apte, G.E. Batley, G.E. Gadd, P.S.
[25] J. Sa, C.A. Aguera, S. Gross, J.A. Anderson, Appl. Catal., B 85 Casey, Environ. Sci. Technol. 41 (2007) 8484.
(2009) 192. [62] J.J. Scott-Fordsmand, P.H. Krogh, M. Schaefer, A. Johansen,
[26] S. Rengaraj, X.Z. Li, Chemosphere 66 (2007) 930. Ecotoxicol. Environ. Saf. 71 (2008) 616.
[27] B.D. Martin, S.A. Parsons, B. Jefferson, Water Sci. Technol. 60 [63] L. Nyberg, R.F. Turco, L. Nies, Environ. Sci. Technol. 42 (2008)
(2009) 2637. 1938.
[28] A. Ghauch, A. Tuqan, H.A. Assia, Environ. Pollut. 157 (2009) [64] A.D. Ostrowski, T. Martin, J. Conti, I. Hurt, B.H. Harthorn, J.
1626. Nanopart. Res. 11 (2009) 251.
[29] J. Wang, L.Q. Jing, L.P. Xue, Y.C. Qu, H.G. Fu, J. Hazard. Mater. [65] J.K. Jiang, G. Oberdorster, P. Biswas, J. Nanopart. Res. 11 (2009)
160 (2008) 208. 77.
[30] T. Hennebel, H. Simoen, W. De Windt, M. Verloo, N. Boon, W. [66] J.Y. Choi, G. Ramachandran, M. Kandlikar, Environ. Sci. Technol.
Verstraete, Biotechnol. Bioeng. 102 (2009) 995. 43 (2009) 3030.
[31] J.Y. Wang, X.J. Huang, J.C.M. Kao, O. Stabnikova, J. Hazard. [67] O. Choi, Z.Q. Hu, Environ. Sci. Technol. 42 (2008) 4583.
Mater. 144 (2007) 292. [68] G. Bystrzejewska-Piotrowska, J. Golimowski, P.L. Urban, Waste
[32] Z. Xiong, F. He, D.Y. Zhao, M.O. Barnett, Water Res. 43 (2009) Manage 29 (2009) 2587.
5171. [69] S.K. Brar, M. Verma, R.D. Tyagi, R.Y. Surampalli, Waste Manage
[33] X.J. Huang, Y.K. Choi, Sens. Actuators, B 122 (2007) 659. 30 (2010) 504.
[34] S. Hwang, M.C. Lee, W. Choi, Appl. Catal., B 46 (2003) 49. [70] A. Afkhami, R. Norooz-Asl, Colloids Surf., A 346 (2009) 52.
[35] N. Keller, E. Barraud, F. Bosc, D. Edwards, V. Keller, Appl. Catal., B [71] B. Geng, Z.H. Jin, T.L. Li, X.H. Qi, Chemosphere 75 (2009) 825.
70 (2007) 423. [72] K. Jia, B.C. Pan, L. Lv, Q.R. Zhang, X.S. Wang, B.J. Pan, W.M.
[36] V. Nagpal, A.D. Bokare, R.C. Chikate, C.V. Rode, K.M. Paknikar, J. Zhang, J. Colloids Interface Sci. 331 (2009) 453.
Hazard. Mater. 175 (2010) 680. [73] M. Medina, J. Tapia, S. Pacheco, M. Espinosa, R. Rodriguez, J.
[37] N. Chitose, S. Ueta, S. Seino, T.A. Yamamoto, Chemosphere 50 Non-Cryst. Solids 356 (2010) 383.
(2003) 1007. [74] Y.C. Chang, D.H. Chen, J. Colloids Interface Sci. 283 (2005) 446.

http://www.elsevier.com/locate/trac 515
Trends
[75] H.V. Tran, L.D. Tran, T.N. Nguyen, Mater. Sci. Eng. C. 30 (2010)
304.
[76] N. Das, Hydrometallurgy 103 (2010) 180.
[77] M. Namdeo, S.K. Bajpai, Colloids Surf., A 320 (2008) 161.
[78] H. Hildebrand, D. Kuhnel, A. Potthoff, K. Mackenzie, A. Springer,
K. Schirmer, Environ. Pollut. 158 (2010) 65.
[79] A. Jasper, H.H. Salih, G.A. Sorial, R. Sinha, R. Krishnan, C.L.
Patterson, Environ. Eng. Sci. 27 (2010) 85.
[80] N. Wang, L.H. Zhu, D.L. Wang, M.Q. Wang, Z.F. Lin, H.Q. Tang,
Ultrason. Sonochem. 17 (2010) 526.
[81] T.J. Yan, J.L. Long, X.C. Shi, D.H. Wang, Z.H. Li, X.X. Wang,
Environ. Sci. Technol. 44 (2010) 1380.
[82] W. Luo, L.H. Zhu, N. Wang, H.Q. Tang, M.J. Cao, Y.B. She,
Environ. Sci. Technol. 44 (2010) 1786.
[83] R.W.M. Krause, B.B. Mamba, L.N. Dlamini, S.H. Durbach, J.
Nanopart. Res. 12 (2010) 449.
516 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 30, No. 3, 2011
[84] T. Ben-Moshe, I. Dror, B. Berkowitz, Appl. Catal., B 85 (2009)
207.
[85] H.Y. Shu, M.C. Chang, C.C. Chang, J. Hazard. Mater. 167 (2009)
1178.
[86] D.W. Elliott, H.L. Lien, W.X. Zhang, J. Environ. Qual. 37 (2008)
2192.
[87] W.P. Hsieh, J.R. Pan, C.P. Huang, Y.C. Su, Y.J. Juang, Sci. Total
Environ. 408 (2010) 672.
[88] L.X. Yang, S.L. Luo, R.H. Liu, Q.Y. Cai, Y. Xiao, S.H. Liu, F. Su,
L.F. Wen, J. Phys. Chem. C. 114 (2010) 4783.
[89] D.W. Elliott, H.L. Lien, W.X. Zhang, J. Environ. Eng. (Reston, VA)
135 (2009) 317.
[90] A. Garcı́a, R. Espinosa, L. Delgado, E. Casals, E. González, V.
Puntes, C. Barata, X. Font, A. Sánchez, Desalination (2010), in
press (doi:10.1016/j.desal.2010.10.052).