Trends in Analytical Chemistry, Vol. 27, No.

11, 2008 Trends

Fate and toxicity of emerging

pollutants, their metabolites
and transformation products
in the aquatic environment
Marinel la Farré, Sandra Pérez, Lina Kantiani, Damià Barceló

There has been a great effort to study the fate, the occurrence and the The way that organic compounds enter
ecotoxicology of emerging pollutants in the aquatic environment. Recently, the environment depends on their pattern
several articles have focused on degradation products of emerging pollutants of usage and mode of application (e.g.,
and the study of their toxicological effects. disposal of municipal, industrial and agri-
We review the fate and the ecotoxicology of emerging pollutants, espe- cultural wastes, excretion of pharmaceu-
cially focusing on their metabolites and transformation products (TPs) in the ticals and accidental spills). Once in the
aquatic environment, including pharmaceuticals, hormones, perfluorinated environment, they can be widely distrib-
compounds, by-products of drinking-water disinfection, sunscreens or UV uted at some time between their produc-
filters, benzotriazoles and naphthalenic acids. tion through to use and disposal. Because
We describe analytical methodologies for the quantitative analysis of most emerging pollutants are from human
emerging pollutants, their metabolites, and their TPs in sewage and surface use, their emissions are an issue for some
waters, and we give the results of monitoring surveys obtained from the wastewater processes, so the study of the
application of these analytical methodologies. fate of the emerging pollutants in waste-
ª 2008 Elsevier Ltd. All rights reserved. water-treatment plants (WWTP) is
important.
Keywords: Analytical methodology; Aquatic environment; Degradation; Emerging
Once released into the environment,
pollutant; Environmental analysis; Fate; LC-MS; Metabolite; Toxicity; Transformation
emerging pollutants are subject to pro-
product
cesses (e.g., biodegradation, and chemical
1. Introduction and photochemical degradation) that
Marinel la Farré, Sandra Pérez, contribute to their elimination. Depend-
Lina Kantiani, ing on the compartment in which syn-
Emerging pollutants are defined as com-
Department of Environmental
pounds that are not currently covered by thetic chemicals are present in the
Chemistry, IIQAB-CSIC c/ Jordi
Girona, 18-26, 08034 existing water-quality regulations, have environment (e.g., groundwater, surface
Barcelona, Spain not been studied before, and are thought to water and sediment) or in the techno-
be potential threats to environmental eco- sphere (e.g., WWTPs and drinking-water
Damià Barceló* facilities), different transformations can
systems and human health and safety (Ta-
Catalan Institute of Water
ble 1). They encompass a diverse group of take place, sometimes producing products
Research, ICRA C/ Pic de
Peguera, 15, 17003 Girona, compounds, including pharmaceuticals, that can differ in their environmental
Spain drugs of abuse, personal-care products behavior and ecotoxicological profile. For
(PCPs), steroids and hormones, surfactants, example, TPs of some pollutants are of-
perfluorinated compounds (PFCs), flame ten more persistent than their corre-
retardants, industrial additives and agents, sponding parent compounds [2] and
and gasoline additives, as well as their exhibit greater toxicity (e.g., the major
transformation products (TPs) (Table 1). In biodegradation product of nonylphenol
*
addition, three new classes have to be added ethoxylates, nonylphenol, which is much
Corresponding author.
to the list of emerging pollutants: nanom- more persistent than the parent com-
Tel.: +34 934 006 100, Ext 435;
Fax: +34 932 045 904; aterials, 1,4-dioxane and swimming pool pound and can mimic estrogenic prop-
E-mail: dbcqam@iiqab.csic.es disinfection by-products (DBPs) [1]. erties [3]).

0165-9936/$ - see front matter ª 2008 Elsevier Ltd. All rights reserved. doi:10.1016/j.trac.2008.09.010 991
Trends
Table 1. Classes of emerging compounds
Compound
Drugs of abuse
Flame retardants
Gasoline additives
Industrial additives and agents
Personal-care products
Fragrances, insect repellants, soaps, antimicrobials, sun-screen agents
Pharmaceuticals
Analgesics and anti-inflammatory drugs, human and veterinary
antimicrobials, antiepileptics, blood-lipid regulators and psychiatric drugs,
anti-tumoral drugs, cardiovascular drugs (b-blockers) and b2-
symphatomimetics, X-ray contrast agents
Steroids and hormones
Surfactants and surfactant metabolites
New classes
Liquid chromatography combined with mass spec-
trometry (LC-MS) and related techniques, such as ultra-
performance LC-MS (UPLC-MS), have become robust,
sensitive techniques for detecting parent compounds and
TPs at ultra-trace levels in environmental samples [4,5].
Unlike gas chromatography (GC)-MS, LC-MS can deter-
mine polar analytes without the need for previous
derivatization. This advantage of LC-MS is particularly
attractive when simultaneously analyzing compounds
belonging to structurally distinct groups whose deter-
mination by GC-MS would involve more than one
derivatization reaction.
With the advent and availability of recent analytical
instrumentation that aids compound identification
[e.g., LC coupled to ion-trap (IT)-MS or time-of-flight
(ToF)-MS], more degradates and metabolites are being
identified. These two MS techniques provide comple-
mentary data that facilitate structural elucidation of
unknown compounds. For example, the ability to
conduct multiple stages of fragmentation in an IT-MS
system can generate MSn spectra with considerable
amounts of structural information that identifies an
unknown degradate. Further confirmation of the pro-
posed identity or chemical formula of new degradation
products can be achieved by accurate-mass measure-
ments using a ToF-MS system or other high-resolution
mass analyzers. Current bench-top ToF-MS instruments
can now achieve a low-femtomole (fmol)-level sensi-
tivity, high resolving power and mass accuracy. Even
more powerful in terms of confirmatory analysis are
hybrid triple-quadrupole ToF-MS (QqToF-MS) systems
that acquire product-ion spectra with accurate-mass
measurements of product ions (precision in the low-
ppm range) [6–9]. An alternative to ToF instruments is
992 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
Examples
Amphetamine, cocaine, tetrahydrocannabinol
C10–C13 chloroalkanes, hexabromocyclododecane,
polybrominated diphenyl ethers, tetrabromo bisphenol A, tris (2-
chloroethyl)phosphate
Dialkyl ethers, methyl-t-butyl ether
Chelating agents (EDTA), aromatic sulfonates
Benzophenone; N,N-diethyltoluamide; methylbenzylidene, nitro,
polycyclic and macrocyclic musks; triclosan
Acetaminophen, acetylsalicylic acid, diclofenac, diazepam,
carbamazepine, benzafibrate, iopromide, iopamidol
Diethylstilbestrol, estradiol, estriol, estrone
Alkylphenol ethoxylates, alkylphenols (nonylphenol and
octylphenol), alkylphenol carboxylates
Nanomaterials
1,4-dioxane
swimming-pool-disinfection by-products
the recently launched LTQ Orbitrap that combines a
conventional linear IT (LIT-MS) with an Orbitrap mass
analyzer. This system provides outstanding mass
accuracy, mass resolution and reliable high-sensitivity
MSn performance.
Using LC-MS, there has been a vast number of ana-
lytical methods for determination of emerging pollutants
and studies of their occurrence in the environment
published (e.g., pharmaceuticals [9–11], hormones [12–
14], endocrine-disrupting compounds [7,15], PFCs [16–
18,3], drinking-water DBPs [3,19,20], sunscreens and
ultraviolet (UV) filters [21,22], brominated flame retar-
dants [23–28] and benzotriazoles [29–31]).
Recently, several articles focused on degradation
products of emerging products and their toxicological
effects. In this sense, in recent years, different review
papers have been published on analytical methods for
emerging contaminants [10–13], and the occurrence of
these compounds in the environment. However, this is
the first review article devoted to the fate and the eco-
toxicology of emerging pollutants and especially focusing
on their metabolites and TPs in the aquatic environ-
ment.
2. Occurrence and fate of emerging pollutants
Emerging pollutants can reach the environment by
being transported and distributed via different routes.
The physico-chemical properties of chemicals (e.g., water
solubility, vapor pressure and polarity) determine their
behavior in the environment. The major sources of
environmentally-relevant emerging contaminants are
primarily WWTP effluents and secondarily terrestrial
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
run-offs (from roofs, pavements, roads and agricultural
land), including atmospheric deposition.
Veterinary drugs used for treatment and prevention of
diseases in farming are deliberately introduced into the
environment when liquid manure is sprayed on agri-
cultural field. Then, veterinary drugs and their metabo-
lites are prone to contaminate soil and groundwater
(Fig. 1). The transport of veterinary drugs to ground-
water can be through leaching or run-off from livestock
slurries, while sorption of the drug in the soil can delay
its distribution.
Human pharmaceuticals enter aquatic systems after
ingestion and subsequent excretion in the form of the
non-metabolized parent compounds or as metabolites
through WWTPs [14]. If the pharmaceuticals and their
human metabolites are susceptible to passing through
WWTPs, they can then enter to rivers or streams. They
can reach groundwater after leaching. In addition,
pharmaceuticals, can reach surface waters by run-off
from fields treated with digested sludge (see Fig. 1).
PCPs (e.g., fragrances) are discharged through
shower waste. They can then pass through WWTPs
and can reach the environment [15]. Nonylphenol is a
degradation product of 4-nonylphenol ethoxylates (4-
NPEOs). Alkylphenol ethoxylates (AEOs) are non-ionic
surfactants that have been used extensively in cleaning
products and industrial processes. More than 90% of
AEOs produced worldwide are NPEOs. Nonylphenol
and NPEOs are commonly present in the WWTPs due
to their extensive domestic and industrial use. As a
Figure 1. Distribution of synthetic chemicals and main transformations in the environment and the technosphere.
Trends
result, AEOs and nonylphenol are found in water and
suspended particulate material present in freshwater,
marine and estuarine environments and sediment [16–
18,3].
Another relevant group of emerging pollutants com-
prises PFCs [3,19,20]. This class includes not only the
well-known perfluorooctane sulfonate (PFOS) and per-
fluorooctanoic acid (PFOA), but also a large number of
other structurally-related compounds. Concern is grow-
ing about this class of compounds, as studies indicate
that they are toxic, persistent, and bioaccumulative.
Moreover, because perfluorocarbons and perfluorosulf-
onic acids are very stable, they eventually turn up in the
environment, especially in water. Since both groups of
compounds have been manufactured for over 50 years,
water currents have spread them so widely that they are
found in the deep sea and even in the Arctic. In addition,
a new generation of PFCs has been launched to the
market. Table 2 summarizes several examples of their
occurrence in natural waters.
An important aspect of this issue is the transformation
and the fate of these materials in environmental and
biological systems. For example, many of the most
widely-used PFCs are volatile and subject to metabolism
or degradation that leads to the formation of their per-
sistent sulfonate and carboxylic acid forms. The most
fundamental aspects of this issue remain largely un-
known. To understand the global transport of PFCs and
their distribution in the environment, it is crucial to
describe these basic metabolic and physical processes.
http://www.elsevier.com/locate/trac 993
Trends Trends in Analytical Chemistry, Vol. 27, No. 11, 2008

Table 2. Examples of concentrations (lg/L) of perfluorinated compounds measured in the aquatic environment

Type of water Country (and site) PFOS PFOA PFHpA PFNA PFDA Ref.
Wastewater
Effluent USA (New-York) 3–68 58–1050 0–376 0–47 [120]
Effluent USA (Kentucky) 8–993 8.3–334 – 0–15.7 0–201 [121]
Effluent USA (Georgia) 0–70 7–227 – 0–54 0–86 [121]

River
Dalälven Sweden – <0.97 0.36 <0.14 – [122]
Vindelälven Sweden – <0.65 0.2 0.22 – [122]
Elbe Germany – 7.6 2.7 0.27 – [122]
Oder Poland – 3.8 0.73 0.73 – [122]
Vistula Poland – 3.0 0.48 0.36 – [122]
Po Italy – 200 6.6 1.46 – [122]
Danuve Romania/Ukraine – 16.4 0.95 0.27 – [122]
Daugava Letonia – <2.2 0.86 0.36 – [122]
Seine France – 8.9 3.7 1.26 – [122]
Loire France – 3.4 0.90 0.43 – [122]
Thames UK – 23 4.1 0.79 – [122]
Rhine Germany – 12.3 3.3 1.50 – [122]
Guadalquivir Spain – 4.6 1.58 1.02 – [122]
Rhine Germany (Breisach) 26 2 – – – [123]
Rhine Germany (Mainz) 12 3 – – – [123]
Rhine Germany (Ludwigshafen) 5 2 – – – [123]
Ruhr Germany (Duisburg) 5 48 – – – [123]
Ruhr Germany (Scwerte) 14 177 – – – [123]
Elpe Germany (Bestwig) – 1168 – – – [123]
Moehne Germany (Heidelberg) 193 3640 148 – – [123]

Lake
Shihwa Korea 89.11 19.22 2.50 3.26 1.98 [124]
Maggiore Italy 7.8 2.4 2.4 0.6 3.7 [125]
Ontario Canada 3.9 2.6 – 3.1 – [126]
Michigan Canada 3.8 3.4 – – – [126]

The analytical techniques for detecting organic con-
taminants are continually being improved and refined,
thanks especially to MS instrumentation that facilitates
sensitive, selective and reliable determination. However,
there is still a lack of information about the TPs in
WWTPs and the environment.
Matrix effects are a great challenge for the quantita-
tive analysis of environmental samples by LC-MS. Signal
suppression or enhancement can compromise the
accuracy of analytical results in trace analysis of com-
plex environmental samples. In general, the evaluation
of degradation products implies work at ultra-trace lev-
els. However, while matrix effects have been studied for
LC-MS2 applications with electrospray ionization (ESI)
[21,22], there have been relatively few studied to eval-
uate matrix effects using atmospheric pressure chemical
ionization (APCI) as the ion source. However, using this
approach, Zhao et al. [23] determined six neutral phar-
maceuticals, including carbamazepine and its major
metabolite, carbamazepine-10, 11-dihydrodiol.
Due to the dilution and the possible degradation of
emerging pollutants in the environment, low levels can
be expected, so an analyte-preconcentration procedure is
usually necessary in order to achieve desired levels of
analytical sensitivity, often requiring high enrichment
factors (100–10,000). Such enrichment factors for
chemical analysis are usually achieved using solid-phase
extraction.
2.1. Pharmaceuticals, steroids, drugs of abuse
Fig. 2 shows human pharmaceuticals, steroids, drugs of
abuse and their metabolites detected in wastewaters and
surface waters using the combination of solid-phase
extraction and MS techniques.
In influent samples, some metabolites are often more
concentrated than their parent compounds. For exam-
ple, in a study of ibuprofen (IBU), an anti-inflammatory
drug, and its metabolites [carboxyibuprofen (CX-IBU)
and hydroxyibuprofen (OH-IBU)] in influent and effluent
samples from a domestic WWTP (Fig. 2a), CX-IBU was
found to be more concentrated than OH-IBU and IBU in
the influent samples [24]. Taking into account that IBU
is excreted as the parent compound (15%), OH-IBU
(26%) and CX-IBU (43%), similar percentages were
found in influent samples. There were similar findings for
citalopram and its metabolite, desmethylcitalopram, in a
WWTP in Norway, where higher concentration of the
metabolite was reported [25].

994 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008 Trends

Figure 2. Maximum levels of human metabolites detected in the environment. WWi, Influent; WWe, Effluent; RW, River water; SeW, Sea water;
ACF, Aceclofenac; 4OH-ACF, 4Õ-hydroxy-aceclofenac; DCF, Diclofenac; 4OH-DCF, 4Õ-hydroxy-diclofenac [24]; ASA, Acetylsalicylic acid; SA,
Salicylic acid; o-OH-HA, o-hydroxy-hippuric acid; GA, Gentisic acid [31,32]; ATV, Atorvastatin; pOH-ATV, p-hydroxyatorvastatin; oOH-ATV,
o-hydroxyatorvastatin; STV, Simvastatin; HydroxyaSTV, Hydroxyl acid simvastatin [37]; CBZ, Carbamazepine; CBZ-EP, 10,-11-dihydro-10,11-
epoxycarbamazepine; 2OH-CBZ, 2-hydroxycarbamazepine; 3OH-CBZ, 3-hydroxycarbamazepine; 10OH-CBZ, 10 hydroxycarbamazepine;
DiOH-CBZ, 11-dihydro-10,11-epoxycarbamazepine [29]; COC, Cocaine; BE, Benzoylecgonine; NorBE, Norbenzoyllecgonine; NorCOC, Nor-
cocaine; CE, Cocaethylene; MORPH, Morphine; 6AcMOR, 6-acetylmorphine; MOR-3B-D-GLUC, Morphine-3ß-D-glucoronide; AMPH,
Amphetamine; MetAMPH, Methamphetamine; MDA, 3,4-methylenedioxyamphetamine; MDMA, 3,4-methylenedioxymetamphetamine; MDEA,
3,4-methylenedioxyethamphetamine; METH, Methadone; EDDP, 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine; 11-norTHC, 11-nor-9-car-
boxy-D9-tetrahydrocannabinol [44]; THC, Tetrahydrocannabinol; OH-THC, 11-hydroxy-THC; LSD, Lysergic acid diethylamide; NorLSD, Nor-
lysergic acid diethylamide; O-OH-LSD, 2-oxo-3-hydroxy-lysergic acid diethylamide [43]; DZP, Diazepam; dDZP, Desmethyl diazepam; FLX,
Fluoxetine; NorFLX, norfluoxetine [26]; CTP, Citalopram; dCTP, Desmethylcitalopram; DidCTP, Didesmethylcitalopram; SERT, Sertraline;
dSERT, Desmethylsertraline [26]; IBU, Ibuprofen; OH-IBU, Hydroxyl ibuprofen; CX-IBU, Carboxy-hybroprofen [27]; SMX, Sulfamethoxazole;
N4AcSMX, N4-Acetylsulfamethoxazole [33,34]; E1, Estrone; E1-3 S, Estrone-3-sulfate; E1-3 G, Estrone-3-glucuronide; E2, Estradiol; E2-3S, Estra-
diol-3-sulfate; E2-3G, Estradiol-3-glucuronide [39]; E3, Estriol; E3-3S, Estriol-3-sulfate; E3-16G, Estriol 16-glucuronide [40].

In the case of OH-IBU, higher levels of concentration
than its parent compound were detected in the effluent
waters. Zwiener et al. [26] reported that OH-IBU was
formed from IBU under aerobic conditions in activated
sludge. However, this transformation did not account for
more than 10% of ibuprofen concentration, suggesting
that the major amounts in sewage of OH-IBU are from
human excretion.
The concentrations of four metabolites of carbamaze-
pine also increased in effluent samples from a municipal

http://www.elsevier.com/locate/trac 995
Trends
WWTP. Carbamazepine is an anti-epilepsy drug that is
highly metabolized in the human body to form 32
metabolites [27]. Carbamazepine and five studied
metabolites were detected in WWTP influent and effluent
at ng/L levels (Fig. 2a) [28]. The concentrations of
10,11-dihydro-10,11-dihydroxycarbamazepine in influ-
ent and effluent were 1.001 lg/L and 1.081 lg/L,
respectively, being about three times that of carbamaz-
epine. The metabolite concentrations were similar in
influent and effluent samples analyzed, indicating little
removal during sewage treatment. The parent com-
pound showed similar behavior.
The hydroxylated metabolites of carbamazepine occur
primarily in conjugated forms in human fluids, and these
conjugated metabolites could have been transformed to
the free form by microbial activity during wastewater
treatment, as described previously for steroids [29].
Salicylic acid, the major metabolite of acetylsalicylic
acid, was detected in raw sewage of a WWTP with an
average concentration of 5.4 lg/L over six days (Fig. 2a).
Gentisic acid and o-hydroxyhippuric acid were present in
the sewage at average concentrations of 4.6 lg/L and
6.8 lg/L, respectively. In the discharge of the WWTP, no
residues of metabolites of acetylsalicylic acid were
detectable, but the parent compound was present at a
concentration of 0.5 lg/L. Hence, removal rates of the
three investigated metabolites exceeded 98% [30].
By contrast, 20 years ago, high concentrations (2.54–
9.71 lg/L) were found for salicylic acid in the effluent
samples from WWTPs in Kansas City [31].
Farré et al. [32] also found high concentrations of
salicylic acid in influents and effluents of wastewaters
and river waters in Catalonia. It should be noted that, in
these initial studies, most of the plants were working
with primary treatments only.
Regarding antimicrobials, there were reports of the
greater elimination of N4-Acetylsulfamethoxazole
(N4AcSMX), a human metabolite of the antimicrobial
sulfamethoxazole (SMX) in a municipal WWTP in Swit-
zerland [33,34]. The concentration of N4AcSMX was
greater in influent, with smaller amounts found in
effluent (Fig. 2a). By contrast, SMX was found in influent
and effluent at similar or lower concentrations. If the
amount of SMX present as acetyl metabolite was ne-
glected, the elimination of SMX would be underestimated.
By contrast, Hilton et al. [35] reported a high level
(2.2 lg/L) of the metabolite in treated sewage, in which
SMX was not detectable.
Concerning atorvastatin, an anti-lipidic drug, similar
concentrations of atorvastatin and its metabolites were
detected in influent samples (Fig. 2a). However, less than
1 ng/L was detected for atorvastatin and its metabolites
in effluent samples [36].
Simvastatin and its human metabolite, simvastatin
hydroxyl acid, were detected at low ng/L in both effluent
and influent samples.
996 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
More than 40 WWTPs were sampled for detection of
acetylsalicylic acid and its human metabolites in effluent,
influent and surface-water samples [30].
A relevant group of pharmaceuticals is antidepres-
sants. Of the 200 most widely dispensed pharmaceuticals
in 2006, six were antidepressants. Despite this wide-
spread use, there are few analytical methods to detect
antidepressants in environmental matrices, so there is
little information about their distribution and fate in the
environment.
Recently, Schultz et al. [37] developed an LC-ESI-MS2
method with limits of detection (LODs) in the range
0.19–0.45 ng/L. The method was applied to wastewater
effluents. Venlafaxine was the predominant antidepres-
sant observed in wastewater and river water [37]. Vas-
skog et al. [25] determined the concentrations of
fluoxetine (FLX), a serotonin reuptake inhibitor, and its
metabolite norfluoxetine (NorFLX) in sewage and
receiving waters in Norway (Fig. 2). The parent com-
pound and its metabolite were detected in influent and
effluent of the WWTP and the results showed that con-
centrations of FLX and its metabolite were lower in
effluent than influent.
Steroids are excreted in urine of humans as more
hydrophilic glucuronides and sulfates. Fig. 2b shows the
concentrations of free steroids and conjugates in sewage
influent and effluent [38,39]. In influent samples,
estrogen-sulfate conjugates were nearly equal in con-
centration to the free steroids. Concerning the concen-
trations of all, E1-3S was present at the highest
concentration (34 ng/L) compared to the estrogen-glu-
curonide concentrations some 100-fold lower. In effluent
samples, much lower concentrations of conjugates were
detected in treated effluent (0.29–0.72 ng/L), suggesting
that activated sludge treatment removed most of estro-
gen conjugates.
The principle that what goes in must come out can
be a helpful tool in estimating the consumption of
drugs of abuse in areas under investigation. For
example, Italian researchers measured the levels of
benzoylecgonine, the major urinary metabolite of co-
caine, in wastewater from several Italian cities [40,41].
What they found was surprising: cocaine use appeared
to be far more common than public health officials
previously thought.
Few studies determining the fate of drugs of abuse and
their metabolites in wastewaters have been published
(Fig. 2b). Two studies performed in Italian and Spanish
WWTPs showed that the concentrations of drugs and
metabolites were lower in effluents than influents,
probably reflecting extensive degradation or sorption of
these substances in WWTPs [42,43].
Due to use of illicit drugs, cocaine and its metabolites,
amphetamines, morphine and its metabolites, 11-nor-9-
carboxy-D9-tetrahydrocannabinol, and methadone and
its main metabolites are present in substantial amounts
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
in influents and effluents, so they can be confirmed as
widespread environmental contaminants.
2.2. Chlorination and disinfection by-products
Chlorination has been successfully used for the control of
water-borne infectious diseases for more than a century.
However, identification of chlorination by-products
(CBPs) in incidences of potential health hazards has
created a major issue in balancing the toxicodynamics of
the chemical species and the risk from pathogenic mi-
crobes in the supply of drinking water. There has been
epidemiological evidence of close relationship between
exposure to CBPs and adverse outcomes, particularly
cancers of vital organs in human beings. CBPs, their
toxicodynamics and removal from drinking water has
been evaluated by Gopal et al. [44].
Specific DBPs of current interest include iodo acids,
bromonitromethanes, iodine-containing trihalome-
thanes (iodo-THMs), brominated forms of 3-chloro-4
(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), halo-
aldehydes, haloamides, and n-nitrosodimethylamine
(NDMA), which is not brominated, but is classified as a
probable carcinogen [45]. Iodoacetic acid, one of five
iodo acids identified for the first time in chloraminated
drinking water, has recently been shown to be more
genotoxic and cytotoxic to mammalian cells than all
DBPs that have been studied, including regulated halo-
acetic acids (HAAs) and bromate [45]. Iodoacetic acid is
twice as genotoxic as bromoacetic acid, which is the
most genotoxic of the regulated HAAs. Other iodo acids
identified {bromoiodoacetic acid, (Z)-3-bromo-3-iodo-
propenoic acid, (E)-3-bromo-3-iodopropenoic acid, and
(E)-2-iodo-3-methylbutenedioic acid [45]} have been
synthesized and are currently under investigation for
genotoxic and cytotoxic effects.
These iodo acids are of concern not only for their po-
tential health risks but also because early research
indicates that they may be maximized in waters treated
with chloramines. Chloramines are generated from the
reaction of chlorine with ammonia, and it appears that
the length of the contact time with free chlorine (before
ammonia is added to form chloramines) is an important
factor in the formation of iodo acids and iodo-THMs.
Because chlorine competes in reaction to form iodate as
a sink for natural iodide, it is likely that plants with
significant contact time with free chlorine before the
addition of ammonia will not produce substantial levels
of iodo acids or iodo-THMs. In addition, new DBPs con-
tinue to be identified [46].
Malliarou et al. [47] carried out a large survey of
HAAs in UK drinking waters. Means were in the range
35–95 lg/L, with an observed maximum of 244 lg/L. In
two of the three regions investigated, there was a good
correlation between total THMs and total HAAs, and the
ratio of total THMs to total HAAs correlated significantly
with temperature, pH, and free and total chlorine.
Trends
Another study was carried out in Athens, Greece, over
a two-year period [48]. DBPs measured included THMs,
haloacetonitriles, haloketones, chloral hydrate, chloro-
picrin, and nine HAAs. All DBPs were identified in pre-
chlorinated drinking-water samples. The most com-
monly detected DBPs were chloroform, trichloroacetic
acid, dichloroacetic acid, and chloroacetic acid. Annual
mean concentrations were in the range 1.1–61.8 lg/L.
Ates at al. evaluated the occurrence of DBPs in surface
waters in Turkey with low levels of dissolved organic
carbon (DOC) [49]. During rainstorm events, land-sur-
face run-off and resuspension of bottom sediments
caused an increase in THM precursors in rivers. These
precursors, when chlorinated at water-treatment facili-
ties, led to the formation of THMs and hence had an
impact drinking-water resources. Alkhatib et al. evalu-
ated the wet weather potential impact of THM formation
in tributaries to drinking-water reservoirs [50].
2.3. Biocides
Among biocides, triclosan is currently used as antimi-
crobial agent and is a PCP. Methyl-triclosan is a TP of
triclosan, and both have been detected in effluent
samples at levels of 7.3 lg/L and 2 ng/L, respectively, as
well as in effluent samples at levels of 300 ng/L and
7 ng/L, respectively. These samples were taken from two
WWTPs, the river Ruhr and a drinking-water facility in
Germany [51]. Although levels of methyl triclosan were
much less than those of triclosan, methyl-triclosan in the
effluent was generally higher than in the influent, sug-
gesting that it was being formed in the WWTPs [52].
The concentrations for triclosan were in the range
<3–10 ng/L in surface waters, whereas values for me-
thyl-triclosan were approximately 40% of those for tri-
closan. Comparable concentrations were detected for
triclosan and methyl-triclosan in a study performed in
eight WWTPs and the Ebro river in Spain.
2.4. UV filters
There has been increased interest in UV filters used in
sunscreens, cosmetics, and other PCPs due to their
presence in environmental waters, their endocrine and
developmental toxicity [53,54] and estrogenicity [55];
the levels observed in environmental waters [56] were
not far below the doses that cause toxic effects in
animals.
In recent years, analytical methods for environmental
assessment have included these compounds [57–59].
Negreira et al. [60] studied the stability of some UV
filters in chlorinated water and identified several halo-
genated by-products.
2.5. Benzotriazoles
Benzotriazoles contain a five-membered ring with three
nitrogen atoms directly bonded to one another as
substituents on a benzene ring. The compounds called
http://www.elsevier.com/locate/trac 997
Trends
methyl (or tolyl) derivatives have a methyl substituent
on the benzene ring. Many other derivatives are possible
and a number have been used in various applications,
the three primary uses being corrosion inhibitor,
UV-light stabilizer for plastics, and anti-foggant in pho-
tography. Because benzotriazoles are used in large
quantities as a corrosion inhibitor, this is the main way
they contaminate the environment. Benzotriazole and
tolyltriazole are soluble in water, resist biodegradation,
and are only partially removed in wastewater treatment
[11].
Because of their water solubility, LC-MS and LC-MS2
methods have been recently developed for their mea-
surement in environmental waters. Weiss and Reem-
tsma developed an LC-ESI-MS2 method for measuring
benzotriazole and two isomers of tolyltriazole (5- and 4-
tolyltriazole) in environmental waters [61]. Using SPE
for extraction, their method could achieve LODs of
10 ng/L (for groundwater) and 25 lg/L (for untreated
wastewater). They found lg/L levels in municipal
wastewater, and removal in wastewater treatment was
poor, so these compounds were recycled to surface
waters and to drinking-water sources. Of the two tolyl-
triazole isomers, 4-tolyltriazole isomer was more stable
in the environment.
Giger et al. measured benzotriazole and tolyltriazole in
rivers and wastewaters in Switzerland [62]. Benzotria-
zole was found at a maximum of 6.3 lg /L in the Glatt
River, and a mass flow of 277 kg per week was observed
in the Rhine River.
Jochmann et al. developed a method using headspace
solid-phase dynamic extraction (SPDE) with GC-MS to
measure dioxane and other contaminants in water [63].
2.6. Naphthalenic acids
Naphthalenic acids are becoming important emerging
environmental contaminants. Research on naphthalenic
acids was initiated in the oil-sand region in Alberta,
Canada, one of the largest producers of crude oil in the
world. When caustic hot water is used in extracting
crude oil from oil sands, that results in a residual tailing
water that contains clay, sand, and organic compounds
of high polarity and molecular weight. The primary toxic
components of the toxic tailing water have been identi-
fied as oil-sand naphthalenic acids. Naphthalenic acids
are toxic to aquatic organisms, including phytoplankton,
daphnia, fish and mammals, and are also endocrine
disrupting. Clemente and Fedorak have published a
review on occurrence, analysis, toxicity and biodegra-
dation of naphthalenic acids [64].
3. Transformation of emerging pollutants
Emerging pollutants undergo various degrees of trans-
formation in engineered surroundings and the environ-
998 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
ment. Some relatively inert molecules persist in the
environment and are difficult to degrade. These com-
pounds can be toxic and accumulate in food chains to
present additional human and ecological risks. Others
can be degraded to TPs. Degradation processes for syn-
thetic chemicals in natural and engineered surroundings
can attenuate the environmental burden of these xeno-
biotics. Various processes may dilute, transform or
eliminate synthetic chemicals, resulting in a cocktail of
parent compounds and TPs.
In the environment, natural attenuation plays a key
role in decontamination of the compartments. Some
processes do not chemically transform the molecule (e.g.,
dispersion and dilution can effectively decrease the
average concentrations of a compound in the environ-
ment). The removal of contaminants by sorption to
sediment and suspended material can also be significant.
Chemical transformations of the synthetic chemicals
can be biotic or abiotic. For example in surface waters,
contaminants may be susceptible to abiotic transforma-
tion processes (e.g., direct photochemical degradation
and a wide array of indirect photochemical pathways,
including reaction with singlet oxygen (1O2), hydroxyl
radical (dOH), peroxy radicals (dOOR), photo-excited
organic matter, and other reactive species) [65].
Photolytic reactions are often complex and lead to
multiple reaction products. These TPs can be more toxic
than the parent compound [66], retain the properties of
the parent compound (e.g., antibiotic activity) as dem-
onstrated by some dehydrated products of tetracyclines
[67] and photodegradation products of the fluoroquin-
olone antibiotic ofloxacin [68], or lose antimicrobial
activity and toxicity. Direct and indirect photolysis in
sunlit aquatic systems can play a more prominent role
than biodegradation in eliminating those chemicals in
the environment that have survived biological treat-
ments in WWTPs.
Partial or total elimination of emerging pollutants
from the environment by microorganisms has been
studied. Many organic compounds are biodegraded by
organisms that utilize the compounds as an energy
source. Another important biodegradation process is co-
metabolism in which an organic compound is modified
but not utilized for growth [69]. Whereas some com-
pounds can evade photochemical reactions because they
are not exposed to sunlight (e.g., when adsorbed onto
particles or in the subsurface), microbial transformation
processes dominate in removal of these organic com-
pounds. Plants and animals can detoxify or excrete
contaminants after uptake, but accumulation in adipose
tissue or the absence of appropriate enzyme systems
necessary for biotransformation can hamper elimination
of the contaminants. Emerging pollutants can also be
microbially transformed in WWTPs, which are designed
to reduce and to minimize the release of potentially
harmful compounds into the aquatic environment.
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
Biotic processes commonly produce compounds of in-
creased polarity whose different physico-chemical prop-
erties result in distinct environmental behavior. In
WWTPs, microorganisms play an important role in
eliminating emerging pollutants. The biological tank
largely comprises bacteria but it also has important
protozoan flora, so that a synthetic chemical can be
transformed to TPs. Using laboratory-scale batch reac-
tors, some studies have investigated the fate of phar-
maceuticals in WWTPs and identified the TPs (e.g.,
biotransformation of drug iopromide in batch reactors
filled with conventional activated sludge and nitrifying
activated sludge collected from a municipal WWTP
[70]). The concentration of iopromide in the batch
reactors decreased exponentially without any initial de-
lay. By day 2, its level had dropped to almost 50%, while,
after 9 days, the primary degradation reached about
90%. Three metabolites produced upon oxidation of the
primary alcohols (forming carboxylates) on the side
chains of iopromide were identified in a batch reactor
containing a liquid mixture from conventional activated
sludge. Furthermore, one metabolite formed by dehydr-
oxylation at the two side chains was identified in a batch
reactor containing a liquid mixture from nitrifying acti-
vated sludge. The MS2-fragmentation pattern of iopro-
mide and its metabolites revealed that the iodinated ring
remained intact and that minor transformations in the
structure occurred during biodegradation of iopromide.
There is increasing interest on compounds produced
by DBPs and emerging pollutants [e.g., ozonation of
pharmaceutical carbamazepine produced three new
oxidation products: 1-(2-benzaldehyde)-4-hydro-(1H,
3H)- quinazoline-2-one (BQM); 1-(2-benzaldehyde)-(1H,
3H)-quinazoline- 2,4-dione (BQD); and, 1-(2-benzoic
acid)-(1H,3H)-quinazoline-2,4- dione (BaQD)] [71]. In
another example, chlorination of acetaminophen pro-
duced 1,4-benzoquinone and N-acetyl-p-benzoquinone
imine [72] and two other ring chlorination products
(chloro-4-acetamidophenol, and dichloro-4-acetamido-
phenol).
Another work reported that chloroform, 5,6-dichloro-
2-(2,4-dichlorophenoxy)phenol, 4,5-dichloro-2-(2,4-
dichlorophenoxy)phenol, and 4,5,6-trichloro-2-(2,
4-dichlorophenoxy) phenol were formed as by-products
from chlorination of antibacterial agent triclosan in
drinking-water treatment [73]. Triclosan, 5-chloro-2-
(2,4-dichlorophenoxy) phenol, is used in many hand
soaps, as well as other products, so one conclusion of this
work was that use of products containing triclosan can
increase a personÕs annual exposure to chloroform by as
much as 40% above background levels in tap water.
VikeslandÕs research group also investigated the
reactivity of triclosan with monochloramine [74], while
the unexpected products and reaction mechanisms of the
aqueous chlorination of cimetidine were investigated by
Buth et al. [75].
Trends
4. Toxicity
There has recently been a great deal of research focused
on investigating biological effects of new emerging pol-
lutants residues in the environment.
4.1. Pharmaceuticals
Pharmaceuticals have been developed to produce a bio-
logical effect, so their residues, metabolites and degra-
dation products in the environment can cause different
ecotoxicological effects that are difficult to predict,
especially in complex matrices.
Acute toxicity of pharmaceuticals has been assessed
by standard tests according to established guidelines
based on bacteria, algae, invertebrates, and fish. Weeb
et al. compared the acute-toxicity responses in different
trophic level organisms of different classes of compounds,
and noted that antidepressants, antibacterials and anti-
psychotics were the more toxic.
Cleuvers et al. assessed the short-term acute toxicity of
non-steroidal anti-inflammatory drugs (NSAIDs) using
algae and invertebrates [76]. In another example, Fer-
rari et al. [77] found higher sensitivity using phyto-
placton than zooplacton. Farré et al. [32] assessed the
acute toxicity of NSAIDs using marine bacteria Vibrio
fischeri. The acute toxicity of b-blockers has not been
extensively studied, with the exception of propanolol,
which was the most toxic b-blocker identified by Huggett
et al. [78,79]. In another study, Hernando et al. [21]
reported the acute toxicity of metoprolol using Daphnia
magna.
Acute toxicity of lipid regulators has not been exten-
sively reported. Clofibrate showed LC50 of 7.7 mg/L for
fish Gambusia holbrooki.
In summary, as can be seen from the EC50 values,
acute toxicity is unlikely to occur at environmental
concentrations, as concentrations at which acute effects
occur are 100–1000 times higher than residues found in
the aquatic environment. However, these compounds
are found in complex mixtures where different effects are
added, and synergistic or antagonistic effects can be
produced.
However, more relevant are chronic effects of micro
pollutants (e.g., pharmaceuticals) because many aquatic
species are continually exposed over long periods, even
their entire life cycle. However, there has been little
information reported to date on different effects and end
points.
Estrogenicity has been one of the best evaluated ef-
fects, but there have been no reports of life-cycle analy-
ses, except for ethynil-estradiol (EE2) [80–85]. Male fish
were exposed to EE2 at 4 ng/L.
Another study reported vitellogenin (VTG) induction
in fathead minnows with an EC50 value as low as 1 ng/L.
In this case, EE2 was 25–30 times more potent [86,87]
http://www.elsevier.com/locate/trac 999
Trends
and these results were in agreement with other previous
studies.
The effects on reproduction of environmentally rele-
vant mixtures of estrogens on Japanese medaka were
recently reported by Jukosky et al. [88]. The effects of EE2
on hormonal responses and xenobiotic biotransforma-
tion system of Atlantic salmon were studied by Mor-
tensen [89].
Estrogenic exposure affected metamorphosis and al-
tered sex ratios in the northern leopard frog and the
identification of critically vulnerable periods of develop-
ment was studied by Hogan et al [84]. In this work, it
was shown that tadpoles exposed to EE2 during mid-
metamorphosis were delayed in development immedi-
ately following exposure and took 2 weeks longer to
reach metamorphic climax. In the unexposed groups,
there was low proportion of intersex tadpoles and gonads
appeared to be morphologically distinct (male and fe-
male) in all individuals. Tadpoles exposed early in
development displayed a strong female-biased sex ratio
compared to the controls.
With respect to the chronic effect of NSAIDs, they
inhibit synthesis and release of prostaglandins via COX.
NSAIDs are commonly found in the environment be-
cause they are one of the more consumed groups of
drugs. Acetylsalicilic acid affected reproduction of D.
magna at concentrations around 1.8 mg/L. Chronic
toxicity studies of diclofenac demonstrated histopathol-
ogical effects in rainbow trout after 28 days of exposure,
producing renal lesions at 5 lg/L [90] and sub-cellular
effects at 1 lg/L [91].
b-blockers bind to b-adrenergic receptors and block
activation by physiological agonists. These receptors are
located in mammals in many tissues, including the
heart, and blockage causes a decrease in the heart rate
and contraction. b-blockers differ in specificity for the
different sub-types of receptor; some act non-specifically
on b1-receptors and b2-receptors (e.g., propranolol),
while others are specific to the b1-receptor sub-type
(e.g., atenolol). Long-term exposure to propranolol re-
duced reproduction in Ceriodaphnia dubia at 250 lg/L
and in Hyalella azteca at 100 lg/L [78].
Current investigations confirm that there is a lack of
knowledge with respect to environmental risks associ-
ated with drug use. The outcome of an environmental
risk assessment very much depends on data on the
amount of substance consumed, degradation, ecotox-
icity (acute or chronic) to which there has been
exposure and the method used to estimate the contents
in the environment. If there was more knowledge
about the biodegradation of pharmaceutical sub-
stances, metabolites and excipients, and about the use
of drugs in society, there would be better data for
predicting any environmental effects associated with
drugs and then better opportunities for targeting risk-
limitation measures.
1000 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
There has been a small number of studies reporting
the toxicity of metabolites and degradation products of
pharmaceuticals, and emerging substances in general
(see Table 3). The behavior and the fate of pharmaceu-
ticals and their metabolites in the aquatic environment
are not well known. However, some of these metabolites
have been shown to be ubiquitous in the environment.
The rather persistent antiepileptic carbamazepine and
clofibric acid, a metabolite of lipid-lowering agents clo-
fibrate, etofibrate and etofyllin clofibrate, were detected
with few exceptions in effluents, freshwater (rivers and
lakes) and even in seawater [92,93].
Farré et al. [32] reported the acute toxicity of salicylic
acid using V. fischeri as test species. In another pio-
neering study, fluoxetine and sertraline and metabolites
norfluoxetine and desmethylsertraline were detected in
fish sampled from the wild in the U.S.A., therefore
reflecting potential bioaccumulation [94]. Whether the
accumulated levels of 1.6 ng/g fluoxetine and 4.3 ng/g
sertraline found in brain have effects on the nervous
system of fish has yet to be investigated.
Adverse effects of paracetamol are mainly due to for-
mation of hepatotoxic metabolites, primarily N-acetyl-p-
benzoquinone imine, synthesized when the availability
of glutathione is diminished in liver cells. Photo-TPs of
naproxen, for instance, showed higher toxicities than the
parent compound, while genotoxicity was not found
[95]. Bedner et al. [72] studied the transformation of
acetaminophen by chlorination. This study showed that
toxicants 1,4-benzoquinone and N-acetyl-p-benzoqui-
none imine were produced.
There was a comparative ecotoxicological hazard
assessment of b-blockers and their human metabolites
using a mode-of-action-based test battery and a quanti-
tative structure-activity relationship (QSAR) approach
[96].
We studied the ecotoxicological hazard potential of 42
pharmaceuticals from 22 therapeutic groups, including
metabolites formed in humans by treating each parent
drug and its metabolites as a mixture of similarly acting
compounds [97]. On average, 50% of a parent drug was
metabolized, and 70% was excreted with urine, albeit
with large variations among drugs. Metabolism reduced
the toxic potential of all but eight drugs. The subse-
quently modeled risk quotient was mostly below the
threshold of 1. However, ibuprofen and its metabolites in
a mixture could pose an ecotoxicological risk, as could,
possibly, acetylsalicylic acid, bezafibrate, carbamazepine,
diclofenac, fenofibrate, and paracetamol.
The reaction of cimetidine, an over-the-counter ant-
acid, with frequently used disinfectant free chlorine was
investigated by Buth et al. [75]. Cimetidine degraded
rapidly in the presence of excess free chlorine, indicating
that it would probably undergo significant transforma-
tion during wastewater disinfection. The predicted no-
effect concentration (PNEC) of the chlorination products
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008 Trends

Table 3. Examples of ecotoxicological studies on emerging pollutants, metabolites and transformation products

Metabolite or degradation Parent compound Toxicological effect Organism tested Ref.

product
Clofibric acid Clofibrate Redox activity in liver Rainbow trout [127]
microsomes (Oncorhynchus mykiss)

Clofibric acid Clofibrate Oxidative stress Gambusia holbrooki [128]

Clofibric acid Clofibrate Respiratory inhibition Human U-937 cells [129]

Salicylic acid Acetyl salicylic acid Acute toxicity Vibrio fischeri [32]

Norfluoxetine Fluoxetine Acute toxicity Japanese medaka (Oryzias latipes) [130]

Norfluoxetine Fluoxetine Acute toxicity Spirostomum ambiguum,

Thamnocephalus platyurus

Norfluoxetine Fluoxetine Inhibition of P450 enzymes [131]

S-norfluoxetine and R-fluoxetine Fluoxetine Enantiospecific sub-lethal Pimephales promelas and [132]
effects Daphnia magna

N-desiso-propylpropranolol, Propanolol Acute toxicity Spirostomum ambiguum, [133]

hydroxypropranolol, and hydroxy Thamnocephalus platyurus
N-desisopropranolol glycol

Furosemide photo-products Furosemide Toxicity and genotoxicity Bacteria, algae, rotifers and [134]
microcrustaceans

Naproxen photo-products Naproxen Toxicity and genotoxicity Bacteria, algae, rotifers and [95]
microcrustaceans

Salicylic acid Acetyl salicylic acid Acute toxicity Selanastrum capricornutum and [135]
microcrustaceans

1,4-benzoquinone and Cimetidine Toxicity [75]

N-acetyl-p-benzoquinone imine

4-chloro-5-methyl-1H-imidazole Cimetidine Toxicity [75]

Methyl-triclosan Triclosan Acute and chronic toxicity Vibrio fischeri, [101]

Dunaliella tertiolecta,
and Palaemonetes pugio

Methyl-triclosan Triclosan Acute toxicity Vibrio fischeri [100]

Trihalomethanes Disinfection by-products Acute Toxicity Selenastrum capricornutum [136]

Tolyltriazole metabolites Tolyltriazoles Acute Toxicity Upland plants [137]

Perfluorooctane- Perfluorooctanesulfonate (PFOS) Peroxisome proliferation; In vitro rat cells [138]

sulfonamide-based PPAR; Rodent
chemicals hepatocarcinogenesis

was estimated, and 4-hydroxymethyl-5-methyl-1H-
imidazole and 4-chloro-5-methyl-1H-imidazole were
estimated to have lower PNECs than cimetidine.
4.2. Biocides
Biocides comprise another relevant group of organic
pollutants from the ecotoxicological point of view. A
large number of papers have reported environmental
concentrations of triclosan, but few reported ecotoxico-
logical data. Orvos et al. [98] studied the effects on
D. magna and fish, reporting values for 48-h D. magna
EC50 to be 390 lg/L and the 96-h median lethal
concentration values for Pimephales promelas and Lepomis
macrochirus to be 260 lg/L and 370 lg/L, respectively.

http://www.elsevier.com/locate/trac 1001
Trends
The toxicity assessment of the impact of different
pharmaceuticals and PCPs (PPCPs), including antimi-
crobial triclosan, on benthic invertebrates was carried
out by Dussault et al. [99]. In this work, the relative
toxicities of triclosan, the lipid regulator atorvastatin
(ATO), the antiepileptic drug carbamazepine (CBZ), and
the synthetic hormone EE2 were compared, with the
most toxic compound being triclosan.
Two recent articles have evaluated the ecotoxicity of
triclosan and its metabolites. Farré et al., [100] studied
the effect of triclosan and metabolite methyl-triclosan on
the marine bacteria V. fischeri. This paper presented the
toxic synergism between triclosan and linear alkylben-
zene sulfonates (LASs).
In another study [101], the acute aqueous toxicity
values (96-h LC50) were 305 lg/L for adult shrimp,
154 lg/L for larvae, and 651 lg/L for embryos. The
bacterium was more sensitive to triclosan than the grass
shrimp, with a 15-min aqueous IC50 value of 53 lg/L
and a 15-min spiked sediment IC50 value of 616 lg/kg.
The phytoplankton species was the most sensitive species
tested, with a 96-h EC50 value of 3.55 lg/L. Adult grass
shrimp were found to accumulate methyl-triclosan after
14-day exposure to 100 lg/L triclosan, indicating for-
mation of this metabolite in seawater and its potential to
bioaccumulate in higher organisms.
A large number and variety of compounds are formed
in the process of chlorinating drinking water. The classes
of compounds formed include THMs, HAAs, haloaceto-
nitriles, halophenols, and halopropanones. The toxico-
logical properties of these compounds have not been well
characterized.
In addition, a number of investigators have shown
that ketones could increase the hepatotoxicity of halo-
alkanes. The ecological effects of trichloroacetic acid in
the environment have been studied in depth [102–104].
4.3. Benzotriazoles
Benzotriazoles and many other derivatives are possible
toxins, and a number have been used in various appli-
cations, including corrosion inhibitor, UV-light stabilizer
for plastics, and anti-foggant in photography. Benzotri-
azole has been considered of very low toxicity and a low
health hazard to humans. However, benzotriazole
derivatives were reported to be mutagenic in bacterial
systems. In addition, 1-amino benzotriazole is a potent
mechanism-based inhibitor of cytochrome P-450s via a
benzyne intermediate, indicating that benzotriazoles as a
class may interact with P-450s. The P-450s are impor-
tant both for detoxifying a broad range of xenobiotics
and for activating many compounds to carcinogens in
mammalian systems.
Benzotriazole has also been demonstrated to be toxic
in plants. In an innovative work, Cancilla et al. [105]
characterized Microtox-active components from aircraft
de-icing or anti-icing fluids, and the primary Microtox-
1002 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
active fraction was shown to be a mixture of benzotria-
zole and tolyltriazoles.
Pillard et al. compared the toxicity of benzotriazole
with that of two methylbenzotriazole isomers and buty-
lbenzotriazole [106]. Acute toxicity assays were used to
model the response of three common test organisms:
Microtox bacteria (V. fischeri), fathead minnow (Pimep-
hales promelas) and water flea (C. dubia). The response of
all the three organisms varied over two orders of mag-
nitude among all compounds. V. fischeri was more sen-
sitive than either C. dubia or P. promelas to all the test
materials.
Corsi et al. [107] also studied the aquatic toxicity of
nine aircraft de-icer and anti-icer formulations and the
relative toxicity of additive package-ingredients AEOs
and 4,5-methyl-1H-benzotriazoles.
However, relevant chronic effects have been associ-
ated with benzotriazoles. Different studies reported the
ecotoxicological effects using in vitro assays to study the
estrogenic activity of 37 components of commercial
sunscreen lotions [108]. But, a recent article by Harris
et al. [109] showed that in vitro assays conducted using
a recombinant yeast (anti-) estrogen assay indicated that
benzotriazole possessed clear anti-estrogenic properties.
This chemical was approximately 100-fold less potent
than tamoxifen, which was used as a positive control.
However, a subsequent in vivo study, involving analysis
of VTG induction and somatic indices in adult fathead
minnows (P. promelas), exposed to this compound at
concentrations of 10, 100, and 1000 lg/L for two
weeks, showed no evidence of anti-estrogenic activity by
benzotriazole.
4.4. Perfluorinated compounds (PFCs)
The presence of some PFCs (e.g., PFOS) at levels above
the LOD has been demonstrated in almost all organisms
sampled in a global survey as well as in both non-ex-
posed and exposed human populations. PFOS concen-
trations in effluents have been reported as approaching
indicative target values derived from available aquatic
toxicity data. PFOA was found to be weakly carcino-
genic. Different review articles [110–112] showed the
importance of the perfluoroalkylated substances for the
environment and the need to fill the current gaps in
knowledge about their environmental fate and effects.
Few studies have been dedicated the study of com-
bined effects of different PFCs. Scialli et al. [113] studied
combining perfluoroalkane-acid exposure levels for risk
assessment.
Recently, reports have suggested that PFCs may have
endocrine-disrupting effects. Liu et al. [114] developed a
non-competitive enzyme-linked immunosorbent assay
(ELISA) method to investigate estrogenic activities of
selected PFCs using VTG induction in primary cultured
hepatocytes of freshwater male tilapia (Oreochromis nil-
oticus). The estimated 48-h EC50 values for E2, 4-
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008 Trends

Table 4. Summary of ecotoxicological data reported for nanomaterials

Material Test species Effect measured Remarks Measurements Ref.

Lysophophatidyl Daphnia magna Biomodification of a Carbon nanotubes [139]
choline-coated carbon-based nanomaterials that are water-
single-walled through digestion soluble with natural
carbon nanotubes lipid layers can be
ingested and
modified
Double-walled Xenopus laevis Acute toxicity and Concentrations No genotoxicity was [140]
carbon nanotubes genotoxicity measured: 10, 100 observed Acute
and 500 mg/L toxic effects at 10
mg/L
Fullerene C60 Daphnia magna Acute toxicity Solvent (THF) EC50 = 460 lg/L [141]
and ultrasonication (THF) EC50 =
7.9 mg/L
(sonication)
Fullerene C60 Daphnia magna Accumulation of Stirring in Addition of 5–8 mg/ [142]
different pollutants ultra-pure water L of fullerene
produced and
increase in
phenanthrene
toxicity
Fullerene C60 Daphnia magna Physiological changes Solvent (THF) Exposure to 260 lg/L [143]
increased heart rate
of Daphnia magna
Fullerene C60 Daphnia magna Acute toxicity Solvent (THF) EC50 = 0.8 lg/L [144]
and stirring (THF) EC50 >35 mg/
L (stirring)
Fullerene C60 Daphnia magna Hyalella Acute toxicity and Stirring in Fixed exposure [145]
azteca Copepod reproduction ultra-pure water concentrations
tested. For Daphnia
magna, effects were
observed
Fullerene C60 Pseudomona putida Lipid composition and Growth-inhibiting [146]
Bacillus subtilis behavior of bacterial concentrations at:
membrane 0.5 mg/L
Pseudomona putida
0.75 mg/L Bacillus
subtilis
Fullerenes C60, Danio rerio Development of embryos 200 lg/L of C60- and [147]
C70, C60(OH)24 (Zebra fish) and physiological changes C70-induced
malformations,
pericardial edema
and mortality.
C60(OH)24 was less
toxic than C60.
Gold nanoparticles Daphnia magna Intake Retention in gut [148]
Single-walled carbon Rainbow trout Respiratory toxicity, and Sodium dodecyl sulfate Respiratory toxicity [149]
nanotubes physiological effects and possible
neurotoxic effects
Single-walled carbon Amphiascus Mortality, development Oxidation and Mortality of 36% at [150]
nanotubes tenuiremis and reproduction dispersion in water 10 mg/L
Single-walled carbon Danio rerio Development of embryos Several conditions [151]
nanotubes (Zebra fish) were tested
TiO2 nanoparticles Daphnia magna Acute toxicity Ultrasonication Increased [152]
immobility of
Daphnia magna
when exposed to
light
TiO2 nanoparticles Daphnia magna Acute toxicity Shaking TiO2 produced 40% [153]
mortality at 20 mg/L
ZnO nanoparticles Pseudokirchneriella Acute toxicity EC50 = 60 lg/L [147]
subcapitata

http://www.elsevier.com/locate/trac 1003
Trends
nonylphenol (4-NP), PFOS, PFOA and 6:2 fluorotelomer
alcohol (FTOH) were 4.7 · 10 7, 7.1 · 10 6,
1.5 · 10 5, 2.9 · 10 5 and 2.8 · 10 5 M, respectively.
In the time-course study, significant VTG induction took
place at 24 h (E2), 6 h (4-NP), 48 h (PFOS), 48 h
(PFOA), 72 h (4:2 FTOH), 12 h (6:2 FTOH), 72 h (8:2
FTOH), and increased further after 96 h of exposure. Co-
exposure to binary mixtures of individual PFCs and E2
for 48 h significantly inhibited E2-induced hepatocellu-
lar VTG production in a dose-dependent manner except
for 4:2 FTOH. The estimated 48-h IC50 (concentration of
a compound that elicits 50% inhibition of maximally E2-
induced VTG) values for PFOS, PFOA, 6:2 FTOH and 8:2
FTOH were 3.1 · 10 7, 5.1 · 10 7, 1.1 · 10 6 and
7.5 · 10 7 M, respectively.
Biochemical effects of PFOS exposure have mainly
been studied in mammalian model species and infor-
mation about effects on fish species remain generally
scarce. This lack of toxicity data highlights that there is
an urgent need to understand the mode of action of this
pollutant at the molecular level. Hagenaars [115]
exposed for 14 days common carp (Cyprinus carpio) to
PFOS at concentrations of 0.1, 0.5 and 1 mg/l in water.
The study of liver tissue using DNA microarray
supported the prediction that increased expenditure of
energy negatively affected processes vital to the survival
of an organism, such as growth.
In a recent study, Shi et al. [116] exposed zebrafish
embryos to investigate the developmental toxicity of
PFOS. Four-hour post-fertilization (4-hpf) zebrafish
embryos were exposed to 0.1, 0.5, 1, 3 and 5 mg/L of
this compound. Hatching was delayed and hatching rates
as well as larval survivorship were significantly reduced
after the embryos were exposed to 1, 3 and 5 mg/L PFOS
for 132 hpf. The fry displayed gross developmental mal-
formations, upon exposure to PFOS concentrations of 1
mg/L or greater. Growth (body length) was significantly
reduced in the 3-mg/L and 5-mg/L PFOS-treated groups.
To test whether developmental malformation was
mediated via apoptosis, flow-cytometry analysis of DNA
content, acridine orange staining and terminal dUTP
nick-end labeling (TUNEL) assay were used. These tech-
niques indicated that more apoptotic cells were present in
the PFOS-treated embryos than in the control embryos.
The overall results indicated that zebrafish embryos
constituted a reliable model for testing the developmental
toxicity of PFCs, and the gene-expression patterns in the
embryos were able to reveal some potential mechanisms
of developmental toxicity.
4.5. Engineered nanomaterials
In recent years, a fair amount of work has been done to
assess the ecotoxicological risk of new engineered
nanomaterials in aquatic environments, but this re-
search is still at an initial stage of development and
several issues need to be resolved.
1004 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
Some controversies and inconsistencies between the
different authors have arisen from some specific prop-
erties of nanomaterials. For example, some nanomate-
rials are almost impossible to disperse in water by
physical methods (e.g., sonication and stirring) and may
require the use of a dispersing agent. Furthermore, the
chemistry outlined above suggests that nanomaterials
will aggregate in natural waters and it could be con-
sidered that it is more ecologically relevant to use the
naturally aggregated nanomaterials for regulatory test-
ing. Another critical point is the chemical characteriza-
tion of the test materials. Preliminary toxicological data
and predicted concentrations, which suggest that
nanomaterials are not acute toxicants, mean that there
is a need for warnings about possible sub-lethal and
long-term effects, and possible synergistic effects with
other toxic pollutants present in the same environmental
compartments. Table 4 summarizes ecotoxicological
effects of new engineered nanomaterials.
5. Conclusions
Surveys of contaminants in the environmental samples
have shown a cocktail of synthetic contaminants in the
environment. However, not only the compound in its
unalterated form has to be monitored, but also TPs
formed in natural or engineered settings need to be taken
into consideration. There is a lack of ecotoxicological
information about some groups of emerging pollutants,
but there is particularly a lack of knowledge of the effects
that can be produced in the receiving environments by
their metabolites and TPs.
Some groups of emerging pollutants (e.g., pharma-
ceuticals) have been studied more in recent years be-
cause they have been produced to have biological effects.
However, in these cases, there is much less information
available about their degradates and metabolites, even
when, for some pharmaceuticals and steroids (e.g.,
citalopram, ibuprofen, diazepam, acetyl salicylic acid and
some estrogens), levels of their metabolites in the envi-
ronment have been reported to be higher than those of
the parent compounds. When aiming to construct a
more comprehensive picture of the contamination sce-
nario, the determination of human metabolites is crucial.
PFCs have received considerable attention, especially
the first PFCs (e.g., PFOS and PFOA). Some studies have
reported toxicity of PFOA and other surfactants
[111,117]. There have also been reviews published on
PFOS, PFOA, and other fluorinated surfactants. Kennedy
et al. wrote a review on the toxicology of PFOA, in which
they discussed the different mechanisms involved in the
types of tumors observed in animal studies [117,118].
Lau et al. reviewed the developmental toxicity of PFOA
and PFOS [111]. There is a need to elucidate transport
processes and the development of new methods for effi-
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
ciently treating wastewaters, containing PFOA and
PFOS. Research is also extending beyond these two
contaminants to other long-chain perfluorinated acids
and alcohols [119].
Another important group of compounds from the
ecotoxicological point of view of their metabolites and
degradation products comprises DBPs. New efforts are
focusing on joint chemical and toxicological evaluation
of mixtures of DBPs produced by different water-treat-
ment processes, with toxicological evaluation focusing
on reproductive and developmental end points, with
assays for other important end points and target organs
(e.g., mutagenicity, carcinogenicity, hepatotoxicity,
nephrotoxicity, immunotoxicity, neurotoxicity, develop-
mental neurotoxicity) and pharmacokinetics.
Benzotriazoles have been studied little to date, but this
group of complexing agents is water soluble, so their
aquatic toxicity causes concern.
In addition, naphthalenic acids are becoming a new
group of water contaminants. It has been shown that
naphthalenic acid is toxic to aquatic organisms, but
ecotoxicological studies of these compounds and other
related substances are at an early stage.
Similarly, research on nanomaterials is at an initial
stage, although research on ecological fate, transport
and health effects is growing rapidly, and to date con-
tinues to be necessary in defining scenarios of exposure.
Research on establishing appropriate ecotoxicity-test
strategies and methods should first define realistic con-
ditions and then test ecotoxicity under these scenarios.
They should consider the fate and the behavior of
nanomaterials in the aquatic environment, with or
without the presence of natural and anthropogenic
substances, and conditions that may influence the
aggregation state. In addition, the formation of aggre-
gates in water offers the opportunity for other organic
materials including toxicants to become associated with
the aggregates, and that will change bioavailability of
these materials and create additional toxicological con-
cerns. Other critical issues are, for example, dispersion of
nanomaterials in water by physical methods (e.g., soni-
cation and stirring) and the chemical assessment of these
materials in environmental samples.
Acknowledgements
The work described in this article was supported by the
Spanish Ministerio de Educación y Ciencia Project
CEMAGUA, and by the European Union through project
NORMAN (Contract No. 018486). Marinel la Farré
thanks the Ministerio de Educación y Ciencia for support
through the I3 P program. This work reflects only the
authorsÕ views, and the European Community is not
liable for any use that may be made of the information
contained therein.
Trends
References
[1] S.D. Richardson, M.J. Plewa, E.D. Wagner, R. Schoeny, D.M. De
Marini, Mutat. Res. 636 (2007) 178.
[2] A.B.A. Boxall, C.J. Sinclair, K. Fenner, D. Kolpin, S.J. Maund,
Environ. Sci. Technol. 38 (2004) 368A.
[3] K. Jahan, R. Ordóñez, R. Ramachandran, S. Balzer, M. Stern,
Water Air Soil Pollut. 8 (2008) 395.
[4] G. Hopfgartner, E. Varesio, V. Tschappat, C. Grivet, E. Bourgogne,
L.A. Leuthold, J. Mass Spectrom. 39 (2004) 845.
[5] F. Hernández, O.J. Pozo, J.V. Sancho, F.J. López, J.M. Marı́n, M.
Ibañez, Trends Anal. Chem. 24 (2005) 596.
[6] S. Pérez, P. Eichhorn, D. Barcelo, Anal. Chem. 79 (2007) 8293.
[7] M. Petrovic, M. Gros, D. Barcelo, J. Chromatogr., A 1124 (2006)
68.
[8] M. Farre, M. Gros, B. Hernandez, M. Petrovic, P. Hancock, D.
Barcelo, Rapid Commun. Mass Spectrom. 22 (2008) 41.
[9] S. Marchese, A. Gentili, D. Perret, G. DÕAscenzo, F. Pastori, Rapid
Commun. Mass Spectrom. 17 (2003) 879.
[10] S.D. Richardson, Anal. Chem. 76 (2004) 3337.
[11] S.D. Richardson, T.A. Ternes, Anal. Chem. 77 (2005) 3807.
[12] M. Petrovic, M.D. Hernando, M.S. Dı́az-Cruz, D. Barceló, J.
Chromatogr., A 1067 (2005) 1.
[13] M.J. Lopez De Alda, S. Dı́az-Cruz, M. Petrovic, D. Barceló, J.
Chromatogr., A 1000 (2003) 503.
[14] S. Pérez, D. Barceló, Trends Anal. Chem. 26 (2007) 494.
[15] A.M. Peck, Anal. Bioanal. Chem. 386 (2006) 907.
[16] M. Farre, M.-J. Garcı́a, M. Castillo, J. Riu, D. Barcelo, J. Environ.
Monit. 3 (2001) 232.
[17] M.A. Blackburn, M.J. Waldock, Water Res. 29 (1995) 1623.
[18] R.A. Rudel, D.E. Camann, J.D. Spengler, L.R. Korn, J.G. Brody,
Environ. Sci. Technol. 37 (2003) 4543.
[19] M.J. Strynar, A.B. Lindstrom, Environ. Sci. Technol. 42 (2008)
3751.
[20] X. Ju, Y. Jin, K. Sasaki, N. Saito, Environ. Sci. Technol. 42 (2008)
3538.
[21] M.D. Hernando, M. Petrovic, A.R. Fernández-Alba, D. Barceló, J.
Chromatogr., A 1046 (2004) 133.
[22] C.R. Mallet, Z. Lu, J.R. Mazzeo, Rapid Commun. Mass Spectrom.
18 (2004) 49.
[23] X. Zhao, C.D. Metcalfe, Anal. Chem. 80 (2008) 2010.
[24] S. Weigel, U. Berger, E. Jensen, R. Kallenborn, H. Thoresen, H.
Hühnerfuss, Chemosphere 56 (2004) 583.
[25] T. Vasskog, T. Anderssen, S. Pedersen-Bjergaard, R. Kallenborn,
E. Jensen, J. Chromatogr., A 1185 (2008) 194.
[26] C. Zwiener, S. Seeger, T. Glauner, F.H. Frimmel, Anal. Bioanal.
Chem. 372 (2002) 569.
[27] K. Lertratanangkoon, M.G. Horning, Drug Metab. Dispos. 10
(1982) 1.
[28] X.S. Miao, B.G. Koenig, C.D. Metcalfe, J. Chromatogr., A 952
(2002) 139.
[29] M. Stumpf, T.A. Ternes, R.D. Wilken, V.R. Silvana, W. Bau-
mann, Sci. Total Environ. 225 (1999) 135.
[30] T.A. Ternes, Water Res. 32 (1998) 3245.
[31] C. Hignite, D.L. Azarnoff, Life Sci. 20 (1977) 337.
[32] M. Farree, I. Farrer, A. Ginebreda, M. Figureas, L. Olivella, L.
Tirapu, M. Vilanova, D. Barcelo, J. Chromatogr., A 938 (2001)
187.
[33] A. Göbel, C.S. McArdell, A. Joss, H. Siegrist, W. Giger, Sci. Total
Environ. 372 (2007) 361.
[34] A. Göbel, C.S. McArdell, M.J.F. Suter, W. Giger, Anal. Chem. 76
(2004) 4756.
[35] M.J. Hilton, K.V. Thomas, J. Chromatogr., A 1015 (2003) 129.
[36] B.J. Vanderford, R.A. Pearson, D.J. Rexing, S.A. Snyder, Anal.
Chem. 75 (2003) 6265.
[37] M.M. Schultz, E.T. Furlong, Anal. Chem. 80 (2008) 1756.
http://www.elsevier.com/locate/trac 1005
Trends
[38] S. Reddy, C.R. Iden, B.J. Brownawell, Anal. Chem. 77 (2005)
7032.
[39] G. DÕAscenzo, A. Di Corcia, A. Gentili, R. Mancini, R. Mastro-
pasqua, M. Nazzari, R. Samperi, Sci. Total Environ. 302 (2003)
199.
[40] E. Zuccato, C. Chiabrando, S. Castiglioni, D. Calamari, R.
Bagnati, S. Schiarea, R. Fanelli, Environ. Health 4 (2005).
[41] E. Zuccato, S. Castiglioni, R. Bagnati, C. Chiabrando, P. Grassi, R.
Fanelli, Water Res. 42 (2008) 961.
[42] S. Castiglioni, R. Bagnati, D. Calamari, R. Fanelli, E. Zuccato, J.
Chromatogr., A 1092 (2005) 206.
[43] C. Postigo, M.J. Lopez de Alda, D. Barcelo, Anal. Chem. 80
(2008) 3123.
[44] K. Gopal, S.S. Tripathy, J.L. Bersillon, S.P. Dubey, J. Haz. Mater.
140 (2007) 1.
[45] S.D. Richardson, Global NEST J. 7 (2005) 43.
[46] H. Gong, Z. You, Q. Xian, X. Shen, H. Zou, F. Huan, X. Xu,
Environ. Sci. Technol. 39 (2005) 7499.
[47] E. Malliarou, C. Collins, N. Graham, M.J. Nieuwenhuijsen, Water
Res. 39 (2005) 2722.
[48] S.K. Golfinopoulos, A.D. Nikolaou, Desalination 176 (2005) 13.
[49] N. Ates, S.S. Kaplan, E. Sahinkaya, M. Kitis, F.B. Dilek, U. Yetis, J.
Haz. Mater. 142 (2007) 526.
[50] E. Alkhatib, R. Peters, Environ. Monit. Assess. 139 (2008) 173.
[51] K. Bester, Arch. Environ. Contam. Toxicol. 49 (2005) 9.
[52] A. Lindstrom, I.J. Buerge, T. Poiger, P.A. Bergqvist, M.D. Muller,
H.R. Buser, Environ. Sci. Technol. 36 (2002) 2322.
[53] D.W. Kolpin, E.T. Furlong, M.T. Meyer, E.M. Thurman, S.D.
Zaugg, L.B. Barber, H.T. Buxton, Environ. Sci. Technol. 36
(2002) 1202.
[54] C. Schmitt, M. Oetken, O. Dittberner, M. Wagner, J. Oehlmann,
Environ. Pollut. 152 (2008) 322.
[55] P.Y. Kunz, K. Fent, Toxicol. Appl. Pharmacol. 217 (2006) 86.
[56] P. Cuderman, E. Heath, Anal. Bioanal. Chem. 387 (2007) 1343.
[57] R. Rodil, J.B. Quintana, P. Lopez-Mahia, S. Muniategui-Lorenzo,
R. Prada, Anal. Chem. 80 (2008) 1307.
[58] B. Kasprzyk-Hordern, R.M. Dinsdale, A.J. Guwy, Water Res. 42
(2008) 3498.
[59] R. Rodil, M. Moeder, J. Chromatogr., A 1179 (2008) 81.
[60] N. Negreira, P. Canosa, I. Rodrı́guez, M. Ramil, E. Rubı́, R. Cela,
J. Chromatogr., A 1178 (2008) 206.
[61] S. Weiss, T. Reemtsma, Anal. Chem. 77 (2005) 7415.
[62] W. Giger, C. Schaffner, H.P.E. Kohler, Environ. Sci. Technol. 40
(2006) 7186.
[63] M.A. Jochmann, M.P. Kmiecik, T.C. Schmidt, J. Chromatogr., A
1115 (2006) 208.
[64] J.S. Clemente, P.M. Fedorak, Chemosphere 60 (2005) 585.
[65] M.W. Lam, K. Tantuco, S.A. Mabury, Environ. Sci. Technol. 37
(2003) 899.
[66] M. Della Greca, A. Fiorentino, M.R. Iesce, M. Isidori, A. Nardelli,
L. Previtera, F. Temussi, Environ. Toxicol. Chem. 22 (2003) 534.
[67] B. Halling-Sorensen, G. Sengelov, J. Tjornelund, Arch. Environ.
Contam. Toxicol. 42 (2002) 263.
[68] J. Sunderland, C.M. Tobin, L.O. White, A.P. MacGowan, A.J.
Hedges, Drugs 58 (1999) 171.
[69] N. Nyholm, F. Ingerslev, U.T. Berg, J.P. Pedersen, H. Frimer-
Larsen, Chemosphere 33 (1996) 851.
[70] S. Pérez, P. Eichhorn, M.D. Celiz, D.S. Aga, Anal. Chem. 78
(2006) 1866.
[71] D.C. McDowell, M.M. Huber, M. Wagner, U. Von Gunten, T.A.
Ternes, Environ. Sci. Technol. 39 (2005) 8014.
[72] M. Bedner, W.A. MacCrehan, Environ. Sci. Technol. 40 (2006)
516.
[73] K.L. Rule, V.R. Ebbett, P.J. Vikesland, Environ. Sci. Technol. 39
(2005) 3176.
[74] A.E. Greyshock, P.J. Vikesland, Environ. Sci. Technol. 40 (2006)
2615.
1006 http://www.elsevier.com/locate/trac
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
[75] J.M. Buth, W.A. Arnold, K. McNeill, Environ. Sci. Technol. 41
(2007) 6228.
[76] M. Cleuvers, Ecotoxicol. Environ. Saf. 59 (2004) 309.
[77] B. Ferrari, R. Mons, B. Vollat, B. Fraysse, N. Paxéus, R.L. Giudice,
A. Pollio, J. Garric, Environ. Toxical. Chem. 23 (2004) 1344.
[78] D.B. Huggett, B.W. Brooks, B. Peterson, C.M. Foran, D. Schlenk,
Arch. Environ. Contam. Toxicol. 43 (2002) 229.
[79] S.F. Owen, E. Giltrow, D.B. Huggett, T.H. Hutchinson, J. Saye,
M.J. Winter, J.P. Sumpter, Aquat. Toxicol. 82 (2007) 145.
[80] R. Länge, T.H. Hutchinson, C.P. Croudace, F. Siegmund, H.
Schweinfurth, P. Hampe, G.H. Panter, J.P. Sumpter, Environ.
Toxicol. Chem. 20 (2001) 1216.
[81] J.L. Parrott, B.R. Blunt, Environ. Toxicol. 20 (2005) 131.
[82] H. Xu, J. Yang, Y. Wang, Q. Jiang, H. Chen, H. Song, Aquat.
Toxicol. 88 (2008) 1.
[83] D.R. Ekman, Q. Teng, D.L. Villeneuve, M.D. Kahl, K.M. Jensen,
E.J. Durhan, G.T. Ankley, T.W. Collette, Environ. Sci. Technol.
42 (2008) 4188.
[84] N.S. Hogan, P. Duarte, M.G. Wade, D.R.S. Lean, V.L. Trudeau,
Gen. Comp. Endocrinol. 156 (2008) 515.
[85] A. Cevasco, R. Urbatzka, S. Bottero, A. Massari, F. Pedemonte,
W. Kloas, A. Mandich, Comp. Biochem. Physiol., C: Pharmacol.,
Toxicol. Endocrinol 147 (2008) 241.
[86] J.V. Brian, C.A. Harris, M. Scholze, A. Kortenkamp, P. Booy, M.
Lamoree, G. Pojana, N. Jonkers, A. Marcomini, J.P. Sumpter,
Environ. Sci. Technol. 41 (2007) 337.
[87] J.V. Brian, C.A. Harris, M. Scholze, T. Backhaus, P. Booy, M.
Lamoree, G. Pojana, N. Jonkers, T. Runnalls, A. Bonfa, A.
Marcomini, J.P. Sumpter, Environ. Health Perspect. 113 (2005)
721.
[88] J.A. Jukosky, M.C. Watzin, J.C. Leiter, Aquat. Toxicol. 86 (2008)
323.
[89] A.S. Mortensen, A. Arukwe, Aquat. Toxicol. 85 (2007) 113.
[90] N. Schlesinger, Drugs 64 (2004) 2399.
[91] R. Triebskorn, H. Casper, A. Heyd, R. Eikemper, H.R. Köhler, J.
Schwaiger, Aquat. Toxicol. 68 (2004) 151.
[92] H.R. Buser, T. Poiger, M.D. Müller, Environ. Sci. Technol. 32
(1998) 3449.
[93] S. Weigel, J. Kuhlmann, H. Hühnerfuss, Sci. Total Environ. 295
(2002) 131.
[94] B.W. Brooks, C.K. Chambliss, J.K. Stanley, A. Ramirez, K.E.
Banks, R.D. Johnson, R.J. Lewis, Environ. Toxicol. Chem. 24
(2005) 464.
[95] M. Isidori, M. Lavorgna, A. Nardelli, A. Parrella, L. Previtera, M.
Rubino, Sci. Total Environ. 348 (2005) 93.
[96] B.I. Escher, N. Bramaz, P. Quayle, S. Rutishauser, E.L.M.
Vermeirssen, J. Environ. Monit. 10 (2008) 622.
[97] J. Lienert, K. Güdel, B.I. Escher, Environ. Sci. Technol. 41 (2007)
4471.
[98] D.R. Orvos, D.J. Versteeg, J. Inauen, M. Capdevielle, A. Rothen-
stein, V. Cunningham, Environ. Toxicol. Chem. 21 (2002) 1338.
[99] E. Dussault, V.K. Balakrishnan, E. Sverko, K.R. Solomon, P.K.
Sibley, Environ. Toxicol. Chem. 27 (2008) 425.
[100] M. Farre, D. Asperger, L. Kantiani, S. Gonzalez, M. Petrovic, D.
Barcelo, Anal. Bioanal. Chem. 390 (2008) 1999.
[101] M.E. DeLorenzo, J.M. Keller, C.D. Arthur, M.C. Finnegan, H.E.
Harper, V.L. Winder, D.L. Zdankiewicz, Environ. Toxicol. 23
(2008) 224.
[102] M.J.M. Wells, L.J. Fono, M.L. Pellegrin, A. Morse, Water Environ.
Res. 79 (2007) 2192.
[103] S.K. Khetan, T.J. Collins, Chem. Rev. 107 (2007) 2319.
[104] T.E. Lewis, T.F. Wolfinger, M.L. Barta, Environ. Int. 30 (2004)
1119.
[105] D.A. Cancilla, A. Holtkamp, L. Matassa, X. Fang, Environ.
Toxicol. Chem. 16 (1997) 430.
[106] D.A. Pillard, J.S. Cornell, D.L. Dufresne, M.T. Hernandez, Water
Res. 35 (2001) 557.
Trends in Analytical Chemistry, Vol. 27, No. 11, 2008
[107] S.R. Corsi, S.W. Geis, J.E. Loyo-Rosales, C.P. Rice, Environ. Sci.
Technol. 40 (2006) 7409.
[108] K. Morohoshi, H. Yamamoto, R. Kamata, F. Shiraishi, T. Koda,
M. Morita, Toxicol. Vitro 19 (2005) 457.
[109] C.A. Harris, E.J. Routledge, C. Schaffner, J.V. Brian, W. Giger, J.P.
Sumpter, Environ. Toxicol. Chem. 26 (2007) 2367.
[110] F.M. Hekster, R.W. Laane, P. de Voogt, Rev. Environ. Contam.
Toxicol. 179 (2003) 99.
[111] C. Lau, K. Anitole, C. Hodes, D. Lai, A. Pfahles-Hutchens, J. Seed,
Toxicol. Sci. 99 (2007) 366.
[112] S.A. Beach, J.L. Newsted, K. Coady, J.P. Giesy, Rev. Environ.
Contam. Toxicol. 186 (2006) 133.
[113] A.R. Scialli, A. Iannucci, J. Turim, Regulat. Toxicol. Pharmacol.
49 (2007) 195.
[114] C. Liu, Y. Du, B. Zhou, Aquat. Toxicol. 85 (2007) 267.
[115] A. Hagenaars, D. Knapen, I.J. Meyer, K. van der Ven, P. Hoff, W.
De Coen, Aquat. Toxicol. 88 (2008) 155.
[116] X. Shi, Y. Du, P.K.S. Lam, R.S.S. Wu, B. Zhou, Toxicol. Appl.
Pharmacol. 230 (2008) 23.
[117] M.E. Andersen, J.L. Butenhoff, S.C. Chang, D.G. Farrar, J.
Kennedy, C. Lau, G.W. Olsen, J. Seed, K.B. Wallace, Toxicol.
Sci. 102 (2008) 3.
[118] J. Kennedy, J.L. Butenhoff, G.W. Olsen, J.C. OÕConnor, A.M.
Seacat, R.G. Perkins, L.B. Biegel, S.R. Murphy, D.G. Farrar, Crit.
Rev. Toxicol. 34 (2004) 351.
[119] M.J.A. Dinglasan, Y. Ye, E.A. Edwards, S.A. Mabury, Environ.
Sci. Technol. 38 (2004) 2857.
[120] E. Sinclair, K. Kannan, Environ. Sci. Technol. 40 (2006) 1408.
[121] K.S. Loganathan, E. Sajwan, K.S. Sinclair, K.K. Kannan, Water
Res. 41 (2007) 4611.
[122] M.S. McLachlan, K.E. Holmstrom, M. Reth, U. Berger, Environ.
Sci. Technol. 41 (2007) 7260.
[123] D. Skutlarek, M. Exner, H. Färber, Environ. Sci. Pollut. Res. 13
(2006).
[124] O. Rostkowski, N. Yamashita, I.M.K. So, S. Taniyasu, P.K.S.
Lam, J. Falandysz, K.T. Lee, S.K. Kim, J.S. Khim, S.H. Im, J.L.
Newsted, P.D. Jones, K. Kannan, J.P. Giesy, Environ. Toxicol.
Chem. 25 (2006) 2374.
[125] R. Loos, J. Wollgast, T. Huber, G. Hanke, Anal. Bioanal. Chem.
387 (2007) 1469.
[126] V.I. Furdui, N.L. Stock, D.A. Ellis, C.M. Butt, D.M. Whittle, P.W.
Crozier, E.J. Reiner, S.A. Mabury, Environ. Sci. Technol. 41
(2007) 1554.
[127] F. Gagné, C. Blaise, C. André, Ecotoxicol. Environ. Saf. 64 (2006)
329.
[128] B. Nunes, A.R. Gaio, F. Carvalho, L. Guilhermino, Ecotoxicol.
Environ. Saf. (2008) (doi:10.1016/j.ecoenv.2007.12.006).
[129] R. Dragone, C. Frazzoli, C. Grappelli, L. Campanella, Ecotoxicol.
Environ. Saf. (2008) (doi:10.1016/j.ecoenv.2008.02.011).
Trends
[130] Y. Nakamura, H. Yamamoto, J. Sekizawa, T. Kondo, N. Hirai, N.
Tatarazako, Chemosphere 70 (2008) 865.
[131] M.T. Donato, J.M. Gomez-Lechon, Curr. Enzyme Inhib. 2 (2006)
281.
[132] J.K. Stanley, A.J. Ramirez, C.K. Chambliss, B.W. Brooks,
Chemosphere 69 (2007) 9.
[133] G. Nałecz-Jawecki, T. Wójcik, J. Sawicki, Environ. Toxicol. 23
(2008) 52.
[134] M. Isidori, A. Nardelli, A. Parrella, L. Pascarella, L. Previtera,
Chemosphere 63 (2006) 785.
[135] G.L. Brun, M. Bernier, R. Losier, K. Doe, P. Jackman, H.B. Lee,
Environ. Toxicol. Chem. 25 (2006) 2163.
[136] Y. Liang, H.C. Hong, L.H. Dong, C.Y. Lan, B.P. Han, M.H. Wong,
Arch. Environ. Contam. Toxicol. 54 (2008) 597.
[137] L.C. Davis, S. Castro-Diaz, D. Lupher, L.E. Erickson, Proc.
Convergence 2000, Environ. Pipeline Eng. Conf., ASCE, Kansas
City, MO, USA, 2000, pp. 118-125.
[138] J.M. Shipley, C.H. Hurst, S.S. Tanaka, F.L. DeRoos, J.L. Butenhoff,
A.M. Seacat, D.J. Waxman, Toxicol. Sci. 80 (2004) 151.
[139] A.P. Roberts, A.S. Mount, B. Seda, J. Souther, R. Qiao, S. Lin,
C.K. Pu, A.M. Rao, S.J. Klaine, Environ. Sci. Technol. 41 (2007)
3028.
[140] F. Mouchet, P. Landois, E. Sarremejean, G. Bernard, P.
Puech, E. Pinelli, E. Flahaut, L. Gauthier, Aquat. Toxicol. 87
(2008) 127.
[141] S.B. Lovern, R. Klaper, Environ. Toxicol. Chem. 25 (2006) 1132.
[142] A. Baun, S.N. Sørensen, R.F. Rasmussen, N.B. Hartmann, C.B.
Koch, Aquat. Toxicol. 86 (2008) 379.
[143] S.B. Lovern, J.R. Strickler, R. Klaper, Environ. Sci. Technol. 41
(2007) 4465.
[144] S. Zhu, E. Oberdörster, M.L. Haasch, Mar. Environ. Res. 62
(2006).
[145] E. Oberdörster, S. Zhu, T.M. Blickley, P. Clellan-Green, M.L.
Haasch, Carbon 44 (2006) 1112.
[146] J. Fang, D.Y. Lyon, M.R. Wiesner, J. Dong, P.J.J. Alvarez,
Environ. Sci. Technol. 41 (2007) 2636.
[147] C.Y. Usenko, S.L. Harper, R.L. Tanguay, Toxicol. Appl. Pharma-
col. 229 (2008) 44.
[148] N.M. Franklin, N.J. Rogers, S.C. Apte, G.E. Batley, G.E. Gadd, P.S.
Casey, Environ. Sci. Technol. 41 (2007) 8484.
[149] C.J. Smith, B.J. Shaw, R.D. Handy, Aquat. Toxicol. 82 (2007) 94.
[150] R.C. Templeton, P.L. Ferguson, K.M. Washburn, W.A. Scrivens,
G.T. Chandler, Environ. Sci. Technol. 40 (2006) 7387.
[151] J. Cheng, E. Flahaut, H.C. Shuk, Environ. Toxicol. Chem. 26
(2007) 708.
[152] K. Hund-Rinke, M. Simon, Environ. Sci. Pollut. Res. 13 (2006)
225.
[153] L.K. Adams, D.Y. Lyon, P.J.J. Alvarez, Water Res. 40 (2006)
3527.
http://www.elsevier.com/locate/trac 1007