Science of the Total Environment 505 (2015) 905–926

Contents lists available at ScienceDirect

Science of the Total Environment

journal homepage: www.elsevier.com/locate/scitotenv

Review

Occurrence and removal of transformation products of PPCPs and illicit

drugs in wastewaters: A review
Eleni N. Evgenidou a, Ioannis K. Konstantinou b, Dimitra A. Lambropoulou a,⁎
a
Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece
b
Department of Environmental and Natural Resources Management, University of Patras, Seferi 2, GR 30100 Agrinio, Greece

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Extensive review on occurrence and

fate of TPs of PPCPs and IDs in WWTPs.
• Scarce data exists on TPs for many
groups of PPCPs and IDs.
• Toxicological data covering effects of
TPs on ecosystems are very poor.
• Efficiency of removal of TPs in WWTPs
is largely unknown.
• In general, concentrations of TPs in
effluents were smaller than in influents.

Abbreviations:OH-THC, 11-hydroxy-Δ9-tetrahydrocannabinol; 14-OH-COD, 14-hydroxycodeine;14-OH-CODN, 14-hydroxycodeinone; Cl2MeP, 3,5-dichloro-methylparaben;ClMeP, 3-

chloro-methylparaben; CBX-IBU, carboxy-ibuprofen; COD, codeinone; E2-3S, estradiol-3-sulfate; E1-3S, estrone-sulfate; OH-IBU, hydroxy-ibuprofen; ISO-NPN, Isoneopine; NPN, Neopine;
DP, 1,5-dimethyl-1,2-dehydro-3-pyrazolone; DiOH-CBZ, 10,11-dihydro-trans-10,11-dihydroxy-carbamazepine; 10-OH-CBZ, 10-hydroxy-10,11-dihydrocarbamazepine; 10-OH-AMTP, 10-
hydroxy-amitriptyline; THC-COOH, 11-nor-carboxy-Δ9-tetrahydrocannabinol; 16αOHE1, 16α-hydroxyestrone; AMDOPH, 1-acetyl-1-methyl-2-dimethyloxamoyl-2-phenylhydrazide;
AMPH, 1-acetyl-1-methyl-2-phenylhydrazide; EDDP, 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine; 2-OH-CBZ, 2-hydroxycarbamazepine; 2′-OH-IBU, 2-hydroxy-ibuprofen; 2-N-
gluc-LMTG, 2-N-glucuronide-lamotrigine; 3′-OH-COT, 3′-hydroxycotinine; 3′-OH-COT, 3′-hydroxycotinine; 3-OH-CBZ, 3-hydroxycarbamazepine; 3-OH-DZP, 3-hydroxydiazepam; PDP,
4-(2-methylethyl)-1,5-dimethyl-1,2-dehydro-3-pyrazolone; 4,6-Cl-TCS, 4,5,6-trichloro-2-(2,4-dichlorophenoxy)phenol; 4-Cl-TCS, 4,5-dichloro-2-(2,4-dichlorophenoxy)phenol; 4-AA,
4-aminoantipyrine; 4-DAA, 4-dimethylaminoantipyrine; 4′-OH-ACF, 4′-hydroxy aceclofenac; 4-OH-DCF, 4′-hydroxy diclofenac; 4OHE1, 4-hydroxyestrone; 4-MAA, 4-
methylaminoantipyrine; 6-Cl-TCS, 5,6-dichloro-2-(2,4-dichlorophenoxy)phenol; 5-OH-DCF, 5-hydroxy diclofenac; 6-AM, 6-acetylmorphine; 9ac, 9-hydroxymethyl-10-carbamoylacridan;
ACD, Acridine; ACDN, Acridone; AEC, anhydroecgonine; AME, anhydroecgonine methyl ester; BE, benzoylecgonine; CBZ-Ep, carbamazepine-10,11-epoxide; CLF, clofibric acid; EBE,
cocaethylene or ethylbenzoylecgonine; COT, cotinine; DES-Me-CTL, desmethyl citalopram; DES-Me-DLZ, desmethyl diltiazem; DES-Me-SER, desmethyl sertraline; DES-Me-VNF,
desmethylvenlafaxine; DES-Me-CTL, didesmethyl citalopram; DIC, dihydrocodeine; DXN, Dioxin; ECG, ecgonine; EME, ecgonine methyl ester; OH-BBP, erythro/threo-hydrobupropion;
ERY-H2O, erythromycin-H2O; FEN acid, fenofibric acid; GUA, guanylurea; OH-BBP, hydroxybupropion; Me-TCS, methyl triclosan; METR-OH, metronidazole-OH; ClMeP, monochloro methyl
paraben; MOR-3â-D-gluc, morphine-3â-D-glucuronide; N-DES-Me-VNF, N-desmethyl venlafaxine; 4-FAA, N-formyl-4-amino-antipyrine; N-F-NOR-COT, N-formylnornicotine; NBE,
norbenzoylecgonine; NCO, norcocaine; NOR-COD, norcodeine; NOR-FLX, norfluoxetine; NOR-LSD, norLSD/nor-isoLSD; NOR-MOR, normorphine; NOR-TRP, nortriptyline; NOR-VRP,
norverapamil; o-DES-Me-VNF, o-desmethylvenlafaxine; o-OH-ATV, o-hydroxy atorvastatin; OSL-CAR, oseltamivir carboxylate; OXZ, οxazepam; OXC, oxytetracycline; p-OH-ATV, p-
hydroxy atorvastatin; RLT acid, ritalinic acid; SA, salicylic acid; SMV hydroxy acid, simvastatin hydroxy acid.
⁎ Corresponding author. Tel.: +30 2310997687; fax: +30 2310997799.
E-mail address: dlambro@chem.auth.gr (D.A. Lambropoulou).

http://dx.doi.org/10.1016/j.scitotenv.2014.10.021
0048-9697/© 2014 Elsevier B.V. All rights reserved.
906 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926

a r t i c l e i n f o a b s t r a c t

Article history: Pharmaceuticals and personal care products (PPCPs) along with illicit drugs (IDs) are newly recognized classes of
Received 28 July 2014 environmental pollutants and are receiving considerable attention because of their environmental impacts: fre-
Received in revised form 6 October 2014 quent occurrence, persistence and risk to aquatic life and humans. However, relatively little information is often
Accepted 6 October 2014
available with regard to their possible biotic and abiotic transformation products (TPs). This lack of knowledge
Available online 7 November 2014
has resulted in a substantial amount of ongoing effort to develop methods and approaches that would assess
Editor: D. Barcelo their occurrence, degradability potential elimination mechanisms and efficiencies in sewage treatment plants
as well as environmental and human health risks. In this article, an extensive literature survey was performed
Keywords: in order to present the current stage of knowledge and progress made in the occurrence of TPs of PPCPs and
Transformation products IDs in raw and treated wastewaters. Apart from the TPs resulting from structural transformations of the parent
PPCPs compound in the aquatic environment or in technological treatment facilities (e.g. sewage and drinking water
Illicit drugs treatment plants), free metabolites and drug conjugates formed during human metabolism have also been in-
Occurrence cluded in this review as they are also released into the aquatic environment through wastewaters. Their concen-
Removal
tration levels were reported in influents and effluents of WWTPs, hospital effluents and their removals in the
Wastewaters
treatment plants were discussed. Finally, information on the toxicity of TPs has been compiled when available.
© 2014 Elsevier B.V. All rights reserved.

Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 906
2. Fate in the wastewater treatment plants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 907
3. Analytical strategies identify transformation products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 907
4. Toxicity of TPs of PPCPs and IDs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 908
5. Inputs and occurrence of TPs of PPCPs and IDs in wastewaters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 908
5.1. Antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 911
5.2. Lipid regulating agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 912
5.3. Analgesics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 914
5.4. Antidepressants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 916
5.5. Psychoactive drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 916
5.6. β-Blockers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 917
6. Statins (antilipidemics) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 917
6.1. Antidiabetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 917
6.2. Anesthetics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 917
6.3. Antiviral drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 918
6.4. Proton pump inhibitors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 918
6.5. Anticancer (cytostatic drugs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 918
6.6. X-ray contrast media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 918
6.7. Stimulants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 918
6.8. Estrogens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 919
6.9. Personal care products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 919
6.10. Illicit drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 920
7. Occurrence of TPs of PPCPs and IDs in biosolids and sludge . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 921
8. Concluding remarks and future outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 921
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 922
Appendix A. Supplementary data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 922
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 922

1. Introduction
Pharmaceuticals and personal care products (PPCPs) are of scientific
and public concern as newly recognized classes of environmental
pollutants and are receiving considerable attention with respect to
their environmental fate and toxicological properties over the last
decade. In addition to PPCPs, many, other categories of new and existing
substances become contaminants of emerging concern. Among them, il-
licit drugs (IDs) have been recently also described as a new unexpected
group of water contaminants with potent psychoactive properties and
unknown effects to the aquatic environment. These three groups of
emerging contaminants (ECs) have attracted a broad scientific interest
which is quickly increasing because of their environmental impacts:
frequent occurrence, persistence and risk to aquatic life and humans.
Numerous studies over the past few years showed that many of
these ECs can persist through wastewater treatment plants (WWTPs)
and effluents are normally considered the primary compartment of
concern since they are discharged into water bodies. Consequently, a
vast number of these compounds has been detected in WWTP effluents,
in receiving surface waters, which in some cases are used for drinking
water production and, less frequently, in ground and drinking water
all over the world (Hirsch et al., 1999; Bendz et al., 2005; Castiglioni
et al., 2006; Hummel et al., 2006; Verenitch et al., 2006; Huerta-Fontela
et al., 2007; Terzić et al., 2008; Shao et al., 2009; Nelson et al., 2011).
Although they are present at low concentration levels, many of them
raise considerable ecotoxicological concerns, either as sole compounds
or also when present as components of complex mixtures (Stülten
et al., 2008).
Despite the increasing information that is available for these new
contaminants and the several excellent dedicated reviews that have
been produced in recent years concerning toxicological properties,
environmental occurrence, and removal in WWTPs (Verlicchi et al.,
2012; Jones et al., 2005; Li et al., 2014; Miège et al., 2009; Luo et al.,
2014), relatively little information is often available with regard to
their possible biotic and abiotic TPs, namely, metabolites, conjugates
and degradation products. This lack of knowledge has resulted in
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
a substantial amount of ongoing effort to develop methods and
approaches that might prove useful for identification as well as for
quantification and assessment of TPs of PPCPs and IDs in different envi-
ronmental compartments. In recent years, an increasing number of
research groups are involved on these subjects and more data and
broader knowledge have become available (Lambropoulou and Nollet,
2014). In this context, several dedicated studies performed under
laboratory, pilot and full scale conditions have been reported in the
open literature to assess their occurrence and elimination efficiency
in WWTPs as well as environmental and human health risks (Reddy
et al., 2005; Castiglioni et al., 2006; Lajeunesse et al., 2008).
The importance of the issue is reflected by the fact that many
organizations, agencies and regulatory authorities have been involved
in the development of a national and/or international research strategy
and priorities or even international legally binding tools to assess the
health risks associated with exposure to trace concentrations of
multiple TPs. For example, relevant regulatory assessment schemes
such as the European Agency for the Evaluation of Medicinal Products
(EMEA) (Carlsson et al., 2006) and the Veterinary International
Cooperation on Harmonization of Technical Requirements for Registra-
tion of Veterinary Medicinal Products (VICH, 2000) have published
guidelines about which stable and/or toxic TPs should be included on
the environmental risk assessment (ERA) of the parent compounds.
Furthermore, the persistence and the toxicity of the TPs of chemical
substances should be considered and assessed in terms of toxic effects
and bioaccumulation potential according to the European chemicals
regulation “REACH” (Escher and Fenner, 2011).
In recognition of the growing concern regarding the TPs of PPCPs
and IDs in urban water cycle, a review study was performed in this arti-
cle in order to obtain information on the progress made in their
occurrence in wastewaters, toxicity effects and elimination efficiencies.
TPs arising from biotic and abiotic structural transformations of the
parent compound in the aquatic environment or in technological
treatment facilities (e.g. sewage and drinking water treatment plants),
free metabolites and drug conjugates formed during human metabo-
lism have been included in this review as they are continuously released
into the aquatic environment through wastewaters.
2. Fate in the wastewater treatment plants
WWTPs play an important role in the life cycle of PPCPs, IDs and
their TPs in modern society. The main transport pathways of these
compounds into the environment are via WWTPs where they may be
only partially eliminated. The removal of organic pollutants at WWTPs
is a complex process with many plausible mechanisms. The removal
efficiency can be affected either by the specific treatment processes
employed (biological or chemical) in individual WWTPs or by the phys-
icochemical characteristics of the pollutants treated, such as water solu-
bility, tendency to volatilize, or to adsorb onto activated sludge, and
degradation half-life by abiotic and biotic processes (Gulkowska et al.,
2008; Behera et al., 2011; Luo et al., 2014).
The efficiency of WWTPs for the elimination of PPCPs and their
TPs is under the control or influence of operational and environ-
mental conditions. The most important of the operational conditions
are: (a) the hydraulic retention time (HRT), (b) sludge retention time
(SRT), and (c) biodegradation kinetics (kbiol), while some of the major
environmental conditions are (a) the temperature with lower effi-
ciencies reported during winter periods and colder climates (Vieno
et al., 2005; Gros et al., 2012; Verlicchi et al., 2012), (b) redox conditions
(different removal efficiencies have been observed for anaerobic, and
aerobic conditions), and (c) pH conditions effecting the degradation ki-
netics of the compounds (Suárez et al., 2008; Gros et al., 2012; Verlicchi
et al., 2012).
Regarding the physicochemical characteristics of the PPCPs, IDs and
their TPs that affect their removal by WWTP's processes, the ability to
interact with solid particles is a major factor because it facilitates their
907
removal by physical–chemical (settling, flotation) or biological process-
es (biodegradation). Thus, compounds with low adsorption coefficients
tend to remain in the aqueous phase, which favors their mobility
through the WWTP and into the receiving waters (Ohlenbusch et al.,
2000).
Generally, compounds with logKow values lower than 3.0 are not
expected to be adsorbed significantly to the particles thus exhibiting
low removal efficiencies in the primary treatment (Behera et al.,
2011). On the other hand, compounds with relatively high logKow
values and pKa values below the pH of the wastewater are expected
to be dissociated in the aqueous phase and not bound to the particles
as well (Thomas and Foster, 2005).
Moreover, considering that some TPs preserve their pharmaceutical
action and their parent compounds are designed to resist to microbial
degradations, their abiotic decomposition is also likely to occur in
wastewaters involving direct and indirect photodegradation, hydrolysis
and oxidoreductive reactions (Sammartino et al., 2008; Khetan and
Collins, 2007; Nikolaou et al., 2007; Heberer, 2002).
However, the efficiencies of WWTPs and the specific treat-
ment stages are largely unknown while it is also difficult to discrim-
inate the elimination process that takes place predominately such as
(i) degradation to lower molecular weight compounds, (ii) physical re-
moval by solids (and consequently removal by sludge), and (iii) trans-
formation into conjugates that can be hydrolyzed later and release the
parent compound. In the latter way, conjugates are acting as reservoirs
of drugs from which the target compounds can then be released to the
environment (Bendz et al., 2005).
Overall, the available data highlight difficulties in drawing a clear
conclusion on the fate and removal of each TP, as many TPs showed sig-
nificantly varied removals in different WWTPs (Fig. 1). However, a sim-
ple classification of these compounds is presented in Table S1.
3. Analytical strategies identify transformation products
Undoubtedly, TP identification remains one of the greatest chal-
lenges in environmental analysis. The analytical challenge related to
the identification of TPs of PPCPs in WWTPs is hampered by the unavail-
ability of reference standards. Analytical approaches for TP determina-
tion can be generally classified into three categories: target analysis
(with reference standards), suspect screening (with suspected sub-
stances based on prior information but no reference standards), and fi-
nally nontarget screening (no prior information, no reference standards)
(Zonja et al., 2014; Calza and Debora, 2014; Schymanski et al., 2014;
Chiaia-Hernandez et al., in press). The first approach guarantees the
undoubted confirmation of a TP, and it could be performed with low
resolution tandem MS systems or with full scan-based methods with
HRMS instrumentation. For the second approach specific information
for compounds that are expected to be in the samples (the “suspects”)
should be available for tentative identification. Suspect screening is
limited by the creation of databases as large as possible, with retention
times and accurate HR mass spectra of both molecular and fragment
ions related to TPs. “Suspects” can be screened using the exact mass of
their expected ions, calculated from the molecular formula. Additional
evidence such as isotope pattern match is usually employed for further
confirmation. For example, element information such as the presence of
Cl, Br, or S has been successfully used to restrict possible molecular
formulas due to their distinct isotope signal at M+2 and characteristic
mass defect compared with natural compounds (Chiaia-Hernandez et
al., in press). The last approach, non-target/unknown screening begins
without any a priori information on the underlying compounds.
Hence, a full identification of the nontarget mass is often difficult, with
no guarantee of a successful outcome. Similar to suspect screening,
high accuracy, high resolution and isotopic pattern data, further im-
proves formula assignment to detected masses. A comprehensive target
screening is generally suggested by researchers before suspect and non-
target screening in order to gain sufficient data for secure structure
908 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926

140

100

Removal Efficiency (%)

60

20

-20

-60

-100

E1 E2 Z E1 T A A A A F F M ne E U Ep eP id eP T P O id T U R id A
H H CB H O -A AA FA A AC C -A o B X-IB Z- 2M Ac lM CO DD -H2 Ac -CO -IB -CA L ac S
- O -O H- 2O H-C 4 4- 4- 4-M H- H-D 6 crid B B l F C E Y N R H L T
α α O C C C CL E O S R
16 16 2- 3-
O O O
4- 4-
A ER F -NO O
-F
N

Fig. 1. Box and Whisker graph showing the removal efficiencies (%) of TPs of PPCPs and illicit drugs in WWTPs (symbol ⊕ displays the mean removals) (Behera et al., 2011; Bendz et al.,
2005; Bueno Martínez et al., 2007; Buerge et al., 2008; Cruz-Morato et al., 2013; Gentili et al., 2002; Gómez-Ramos et al., 2011; González-Mariño et al., 2011; Gulkowska et al., 2008;
Huerta-Fontela et al., 2008; Jiang et al., 2005; Karthikeyan and Meyer, 2006; Kosma et al., 2010, 2014; Lee et al., 2005; Letzel et al., 2010; Pérez and Barceló, 2008; Prasse et al., 2010; Roberts and
Thomas, 2006; Rosal et al., 2010; Schlusener, 2005; Stamatis and Konstantinou, 2013; Terzić et al., 2008; Xu et al., 2007).

elucidation and improve identification efforts toward “known un-
known” compounds (Krauss et al., 2010; Schymanski et al., 2014;
Chiaia-Hernandez et al., in press).
A number of excellent works (Krauss et al., 2010; Zonja et al., 2014;
Calza and Debora 2014; Schymanski et al., 2014; Chiaia-Hernandez et
al., in press) giving a comprehensive theoretical background for these
approaches are available in the literature, so such information are not
further discussed. In particular, systematic investigations have been
performed by using target screening analysis in almost all the studies
reviewed herein. Few exceptions come from the works of Ternes'
group and other researchers (Ternes and Hirsch, 2000; Schulz et al.,
2008; Kormos et al., 2009, 2010, 2011; Kosma et al., submitted for
publication) who used suspect screening analysis after the generation
and study of TPs in lab scale experiments. In addition, known urine me-
tabolites have been also included in suspect screening analysis as in the
case of omeprazole (Boix et al., 2014).
4. Toxicity of TPs of PPCPs and IDs
The TPs may preserve the same mode of action as the parent com-
pound if for example the active moiety remains intact after transforma-
tion, thus producing a toxicological effect on non-target organisms in
environmental systems (aquatic or terrestrial) (Boxall et al., 2004). As
the TPs usually exist as a mixture with their parent compound, their
contribution to the overall ecotoxicological effect cannot be neglected.
The toxicity of TPs can differ from the toxicity of their parent compound
in two ways: in their bioconcentration (toxicokinetics) and/or their
mode of toxic action (toxicodynamics). In rare cases, transformation
can create new toxicophores that can lead to higher toxicity by a similar
or dissimilar mode of action (Michael et al., 2014).
Although individual chemicals are typically present at low concen-
trations, they can interact with each other resulting in additive or poten-
tially even synergistic mixture effects. Therefore it is necessary to assess
the environmental risk caused by these substances and data on expo-
sure is required for quantitative risk assessment of TPs and/or metabo-
lites. However, such data are mostly missing because of the non-
availability of TPs and very often metabolites for experimental testing.
In most of studies, the toxicity is generally expressed as an overall tox-
icity of the sample consisting of a mixture of TPs formed and possibly
parent compound. This approach is generally used straightforward in
experiments simulating environmental conditions by performing biotic
(Rubirola et al., 2014) or abiotic transformation studies (photolysis,
ozonation, AOPs etc.) (Fatta-Kassinos et al., 2011) or having a real envi-
ronmental sample. The assessment of toxicity is usually conducted by
using bioassays based on algae, Vibrio fischeri, daphnia and fish organisms.
These tests can only be used as a very first indication without
providing information on potential gene toxicity, neurotoxicity, etc.
(Celiz et al., 2009). Overall, toxicological data covering effects of target
TPs of PPCPs on ecosystems are very poor; in particular, no systematic
investigations have provided figures on their impact on the environ-
ment and consequently, research on this direction should be encourage.
5. Inputs and occurrence of TPs of PPCPs and IDs in wastewaters
After consumption, PPCPs and IDs are primarily excreted in urine
and feces as a mixture of the parent compound and its metabolites.
The metabolites can be categorized as Phase I and Phase II metabolites.
Phase I metabolites stem from in vivo biochemical oxidation, reduction,
and hydrolysis reactions that increase their aqueous solubility and facil-
itate elimination from the body. On the other hand, Phase II metabolites
(usually named as conjugated metabolites) are the result of biochemical
reactions that add a molecule (i.e. glucuronic acid) to the parent com-
pound (Holčapek et al., 2008). Phase II metabolites deconjugate back to
the parent compound in wastewater (D'Ascenzo et al., 2003) and are in-
frequently identified in surface waters (Ferrer and Thurman, 2010).
However, after excretion, both the parent compounds and their metabo-
lites entering the sewage systems and the WWTPs can undergo structur-
al changes by a variety of processes which can be biotic (biodegradation
by bacteria and fungi) and abiotic like hydrolysis or photolysis (Michael
et al., 2014). Consequently, TPs of PPCPs and IDs can stem either from
human metabolism, or from biotic and abiotic processes.
No specific pattern is observed in the occurrence of TPs of PPCPs and
IDs in municipal, pharmaceutical industry and hospital wastewaters
(Tables 1–4, Tables S2–S5; Supplementary material). In addition, great
discrepancies are observed between countries indicating the regional
differences in drug use patterns (Figs. 1 and S1). Variations are also ob-
served within the same country, between different WWTPs (Gulkowska
et al., 2008). This can be attributed to the different pharmaceutical pre-
scription patterns in a country which determine or affect influent con-
centrations, and to the removal efficiencies of individual WWTPs,
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926 909

Table 1
Occurrence of most frequently TPs of antibiotics and analgesics in influents and effluents of WWTPs.

Influents Effluents

TPs Occur. Min Max Mean Ref. Occur. Min Max Mean Ref.

(Parent compound/ Freq.% (ng/L) Freq.% (ng/L)

(excretion unchangeda (%))

Antibiotics
ERY-H2Ob,c (erythromycin/(5–25)) 60 70 1200 340 Karthikeyan and Meyer 59.6 n.d 6000 2500 Miao et al. (2004)
(2006)
– n.d – 1978 Xu et al. (2007) – b20 199 – McArdell et al. (2003)
– n.d 810 470–810 Gulkowska et al. (2008) 30 90 300 270 Karthikeyan and Meyer (2006)
– n.d 850 510–850 Gulkowska et al. (2008) 100 n.d 838 80 Shao et al. (2009)
100 24 420 134 Terzić et al. (2008)
100 n.d 6110 938 Lin and Tsai (2009)
100 n.d 7840 110 Lin and Tsai (2009)
87.5 27 159 – Gómez-Ramos et al. (2011)
OH-METRc (metronidazole/(20)) – n.d 145 62.9 Santos et al. (2013) – 64.7 158 102 Santos et al. (2013)
– n.d 11,344 Santos et al. (2013)
CLF acidb,c (clofibrate/(6)) – n.d 127 26 Rosal et al. (2010) – b6 91 12 Rosal et al. (2010)
20 n.d. 110 – Weigel et al. (2004)
50 n.d 170 – Weigel et al. (2004) 29 n.d – b10 Gómez-Ramos et al. (2011)
73 n.d 10 10 Gómez-Ramos et al. (2011) 100 b20 44 Roberts and Thomas (2006)
100 20 651 – Roberts and Thomas (2006) b30 n.d – b100 Terzić et al. (2008)
– n.d 65 28 Behera et al. (2011) 84.2 7 81 27 Bueno Martínez et al. (2007)
15.6 n.d 265.9 – Kosma et al. (2014) – n.d 6 2 Behera et al. (2011)
100 308 740 527 Stamatis and Konstantinou 9.4 n.d 70.8 – Kosma et al. (2014)
(2013)
77 n.d. 41,428 – Salgado et al. (2011) 100 86.5 258 184 Stamatis and Konstantinou
(2013)
100 63.8 203 135 Stamatis and Konstantinou
(2013)
FEN acidc (fenofibrate/(little)) 100 n.d 117 79 Rosal et al. (2010) 100 b8 129 78 Rosal et al. (2010)
78.9 4 349 186 Bueno Martínez et al. (2007)

Analgesics — NSAIDs
SAb,c (acetylsalyciic acid/(little)) – 1.8 10,800 4900 Ort et al. (2010) 100 554.3 2178.2 – Verenitch et al. (2006)
100 2820 12,700 6860 Lee et al. (2005) 100 10 320 140 Lee et al. (2005)
– n.d 850 – Cruz-Morato et al. (2013) 87.5 n.d 6825 Kosma et al. (2014)
96.9 n.d 89,135 – Kosma et al. (2014) 100 90.2 256 169 Stamatis and Konstantinou
(2013)
100 576 1673 1157 Stamatis and Konstantinou 100 46.8 224 120 Stamatis and Konstantinou
(2013) (2013)
OH-IBUc (ibuprofen/(10)) 100 n.d 990 990 Bendz et al. (2005) 100 n.d – 50 Bendz et al. (2005)
100 1320 6840 – Weigel et al. (2004) 100 90 1130 – Weigel et al. (2004)
– n.d. 200 – Batt et al. (2008)
CBX-IBUc 100 n.d 10,750 10.750 Bendz et al. (2005) 100 n.d – 430 Bendz et al. (2005)
100 1630 23,000 – Weigel et al. (2004) 80 n.d. 1270 Weigel et al. (2004)
c
4-OH-DCF (diclofenac/(5–10)) – n.d 237 237 Pérez and Barceló (2008) – b59 860 (b59–450) Stülten (2008)
4′-OH-ACFc – 43 82 – Pérez and Barceló (2008) – b59 1600 (b59–710) Stülten (2008)
– n.d – 176 Pérez and Barceló (2008)
n.d 104 2007
– b69 660 b69–420 Stülten (2008)
4′-OH-ACFc (aceclofenac (b5)) – n.d – (b10) Pérez and Barceló (2008)
DPc (phenazone/(−)) 40 n.d 70 30 Zuehlke et al. (2004) 100 n.d 110 60 Zuehlke et al. (2004)
AMDOPHb 100 390 710 270–550 Zuehlke et al. (2004) 100 350 730 270–600 Zuehlke et al. (2004)
(dimethylaminophenazone/(−))
AMPHb 92 n.d 150 40–110 Zuehlke et al. (2007) 88 130 190 50–110 Zuehlke et al. (2007)
4-AAc (metamizol/(N90)) – 262 3325 1517 Rosal et al. (2010) – 127 2253 676 Rosal et al. (2010)
63.2 131 9286 2098 Bueno Martínez et al. (2007)
4-MAAc – 314 1894 880 Rosal et al. (2010) – 34 1098 291 Rosal et al. (2010)
78.9 9 9253 1051 Bueno Martínez et al. (2007)
c
4-AAA – 1760 22,200 8333 Rosal et al. (2010) – b100 6745 4489 Rosal et al. (2010)
4-AAAc 100 830 8800 630–6650 Zuehlke et al. (2004) 73.7 2109 25,030 7260 Bueno Martínez et al. (2007)
100 1010 7000 470–5300 Zuehlke et al. (2004)
4-FAAc – 1005 71,000 17,579 Rosal et al. (2010) – b33 27,444 5593 Rosal et al. (2010)
4-FAAc 100 670 1900 320–1300 Zuehlke et al. (2004) 73.7 40 10,114 3386 Bueno Martínez et al. (2007)
100 510 2000 30–1400 Zuehlke et al. (2004)
4-DAAc 5.3 n.d – 122 Bueno Martínez et al. (2007)
PDPc 36 n.d 40 20 Zuehlke et al. (2004) 36 n.d 30 20 Zuehlke et al. (2004)
a
Petrie et al. (2014).
b
Abiotic transformation.
c
Biotic transformation.

which affect effluent concentrations. According to the present literature which is not surprising as these countries are be in command of the en-
survey, most of the studies on TPs of pharmaceuticals have been pro- vironmental research and host researchers with outstanding and distin-
duced conducted in Germany and Spain, followed by USA and Canada, guished expertise in the field. TPs of PCPs and IDs were less studied with
910 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926

Table 2
Occurrence of most frequently TPs of antidepressant, psychoactive and psychostimulant drugs in influents and effluents of WWTPs.

Influents Effluents

TPs Occur. Min Max Mean Ref. Occur. Min Max Mean Ref.

(Parent compound/ Freq.% (ng/L) Freq.% (ng/L)

(excretion unchangeda (%))

Antidepressants
NOR-FLXb (fluoxetine/(11)) 100 n.d 9.9 9.9 Vanderford and Snyder (2006) – n.d – 5 Schultz and Furlong (2008)
NOR-FLXb 100 0.7 9.3 – Vasskog et al. (2008) – det Batt et al. (2008)
– n.d 4.2 1.8–4.2 Lajeunesse et al. (2008) 100 n.d – 3.9 Vanderford and Snyder
(2006)
– n.d 11 n.d.–11 Metcalfe et al. (2010) 100 n.d. 2.4 Vasskog et al. (2008)
– n.d – 1.7–1.8 Lajeunesse et al. (2008)
– n.d – 5 Metcalfe et al. (2010)
b
DES-Me-SER 100 n.d 30.5 – Vasskog et al. (2008) 100 n.d. 10.6 Vasskog et al. (2008)
(sertraline/(trace)) – n.d 21 5.0–21 Metcalfe et al. (2010) – n.d – b13 Metcalfe et al. (2010)
– n.d 5 4.2–5.0 Lajeunesse et al. (2008) – n.d – 3.6–4.7 Lajeunesse et al. (2008)
N-DES-Me-VNFb – n.d 259 134–259 Metcalfe et al. (2010) – n.d – 217 Metcalfe et al. (2010)
(venlafaxin/(5))
O-DES-Me-VNFb – n.d 345 274.3–345.9 Lajeunesse et al. (2008) – n.d – 222.5–330 Lajeunesse et al. (2008)
– n.d 2602 1065–2602 Metcalfe et al. (2010) – n.d – 1637 Metcalfe et al. (2010)
– b10 5500 Writer et al. (2013)
DES-Me-VNFb 16.7 n.d 5 5 Huerta-Fontela et al. (2010) 16.7 n.d – 2 Huerta-Fontela et al. (2010)
DES-Me-CTb (citalopram/(10)) 100 118 426.7 – Vasskog et al. (2008) 100 36.3 300.5 – Vasskog et al. (2008)
– 0 133 104–133 Metcalfe et al. (2010) – n.d – 111 Metcalfe et al. (2010)
100 5.6 20 – Vasskog et al. (2008) – b10 200 – Writer et al. (2013)
Di-DES-Me-CTLb 100 n.d. 10 – Vasskog et al. (2008)
NOR-triptylineb – 3.1 4.5 – Lajeunesse et al. (2008) – 1.5 3.8 – Lajeunesse et al. (2008)
(triptyline/(30–50))
b
10-OH-amitriptyline – n.d. 64 – Batt et al. (2008)
DES-Me-diltiazemb – 65 110 – Batt et al. (2008)
(diltiazem (b4))
NOR-VRPb (verapamil (b4)) – n.d. 71 – Batt et al. (2008)
3-OH-DZPb (diazepam/(trace)) – n.d 320 320 Bones et al. (2007) – n.d.–126 126 – Bones et al. (2007)
72.7 n.d 230 55 Hummel et al. (2006) 100 180 50 Hummel et al. (2006)
Οxazepamb 100 n.d 860 480 Hummel et al. (2006) 100 n.d 630 320 Hummel et al. (2006)
b
OH-BBP (bupropion (b1%)) – b10 2000 – Writer et al. (2013)
Erythro/threo-hydroBBP – b10 5700 – Writer et al. (2013)

Psychoactive drugs
CBZ-Epb (carbamazepine/(3)) 50 n.d 63 – Rosal et al. (2010) 42.1 13 110 52 Bueno Martínez et al. (2007)
100 880 4026 – Huerta-Fontela et al. (2010) 100 69 3581 – Huerta-Fontela et al. (2010)
– 300 500 350 Gómez et al. (2007) – b70 300 160 Gómez et al. (2007)
– n.d 27 Leclercq et al. (2009) – b5.2 29 – Leclercq et al. (2009)
– n.d 39.2 39.2 Miao et al. (2005) – n.d – 19.1 Miao et al. (2005)
– n.d 47.2 47.2 Miao and Metcalfe (2003) – – – 52.3 Miao and Metcalfe (2003)
– 19,820 75,500 – Cruz-Morato et al. (2013) – – – 87 Bahlmann et al. (2014)
– – – 59 Bahlmann et al. (2014) – – – 50 Bahlmann et al. (2014)
– – – 48 Bahlmann et al. (2014)
DiOH-CBZb,c – 264 1460 – Leclercq et al. (2009) – 311 1500 Leclercq et al. (2009)
100 n.d 3700 N2000 Hummel et al. (2006) 100 – 3600 N2000 Hummel et al. (2006)
– n.d 1001.2 1001.2 Miao et al. (2005) – – – 1081.2 Miao et al. (2005)
100 n.d 92 – Hummel et al. (2006) 100 – 90 Hummel et al. (2006)
– n.d 1571.7 1571.7 Miao and Metcalfe (2003) – – – 1325.0 Miao and Metcalfe (2003)
– – – 4000 Bahlmann et al. (2014) – b10 400 Writer et al. (2013)
– – – 1900 Bahlmann et al. (2014) – – – 3400 Bahlmann et al. (2014)
– – – 2000 Bahlmann et al. (2014)
2-OH-CBZb – n.d 39 – Leclercq et al. (2009) – 5 48 Leclercq et al. (2009)
– n.d 59 59 Miao et al. (2005) – – – 70.4 Miao et al. (2005)
– n.d 121 121 Miao and Metcalfe (2003) – – – 132.3 Miao and Metcalfe (2003)
– 500 163,800 – Cruz-Morato et al. (2013)
1/2-OH-CBZb – – – 170 Bahlmann et al. (2014) – – – 140 Bahlmann et al. (2014)
– – – 90 Bahlmann et al. (2014) – – – 97 Bahlmann et al. (2014)
3-OH-CBZb – n.d 55.4 55.4 Miao et al. (2005) – – – 69.2 Miao et al. (2005)
– n.d 94.8 94.8 Miao and Metcalfe (2003) – – – 101.5 Miao and Metcalfe (2003)
– – – 150 Bahlmann et al. (2014) – – – 140 Bahlmann et al. (2014)
– – – 92 Bahlmann et al. (2014) – – – 95 Bahlmann et al. (2014)
9acb – n.d 0 Leclercq et al. (2009) – 4–18% Leclercq et al. (2009)
10-OH-CBZb – n.d 22.2 22.2 Miao et al. (2005) – – – 32.5 Miao et al. (2005)
– n.d 8.5 8.5 Miao and Metcalfe (2003) – – – 9.3 Miao and Metcalfe (2003)
– n.d 1115 – Leclercq et al. (2009) – b10 1900 – Writer et al. (2013)
– – – 490 Bahlmann et al. (2014) – 7.5 1170 – Leclercq et al. (2009)
– – – 270 Bahlmann et al. (2014) – – – 500 Bahlmann et al. (2014)
– – – 270 Bahlmann et al. (2014)
Acridineb,c – n.d 31 – Leclercq et al. (2009) – 5.7 38 – Leclercq et al. (2009)
– n.d 19 – Leclercq et al. (2009)
– n.d 1010 – Cruz-Morato et al. (2013)
Acridoneb – n.d 23 – Leclercq et al. (2009)
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
Table 2 (continued)
Influents
TPs Occur. Min Max Mean Ref.
(Parent compound/ Freq.% (ng/L)
(excretion unchangeda (%))
Psychostimulant drugs
RTL acidb 81.3 50 270 – Letzel et al. (2010)
(methylphenidate/(b1))
a
Petrie et al. (2014).
b
Biotic transformation.
c
Abiotic transformation.
Italy to be a pioneer in the field of IDs (Fig. S2). The most frequently de-
tected TPs of some typical PPCPs (antibiotics, analgesics, lipid regulating
agents, antidepressant psychoactives, estrogens etc.) and IDs will be
next reviewed paying special attention to the key aspects of their occur-
rence in wastewaters in order to address the major questions and con-
cerns relating to their presence in the environment.
5.1. Antibiotics
Most antibiotics are poorly absorbed by humans or animals and con-
sequently 25 to 75% are excreted unaltered through urine and feces
(Chee-Sanford et al., 2001). Vast bibliography exists concerning the
occurrence of antibiotics in wastewaters while on the contrary, the
opposite phenomenon is observed regarding the TPs of antibiotics in
wastewaters. Macrolide antibiotics are an important class of
antibiotics and they are considered as the most important
antibacterial agents used in human medicine. One of the most studied
TPs is erythromycin-H2O (ERY-H2O), the dehydrated metabolite of
ERY, due to the fact that ERY is not stable in the aquatic environment
and it is rapidly converted in the dehydrated product. This dehydration
process is favored at acidic pH and it is not surprising that the parent
compound is almost not detectable in aquatic environment considering
the fact that ERY is medicated orally and thus, has to pass through
strongly acidic conditions in the stomach. Consequently, the detection
of ERY in wastewaters can proceed only via the detection of its
dehydrated metabolite, ERY-H2O. According to the literature, this TP
does no longer exhibit antibiotic properties (Roth and Fenner, 1994).
As indicated in Table 1 (see also Figs. S3 and S5 and Tables S2 and
S4), ERY-H2O was frequently detected in the influents and the effluents
of WWTPs and in many cases the frequency of detection reached 100%.
Moreover, Karthikeyan and Meyer (2006) observed that the detection
of this TP presented seasonal variations and exhibited higher frequen-
cies especially when the samples were collected in the fall.
According to Table 1 the concentrations of ERY-H2O in wastewater
influents go up to 1978 ng/L (Xu et al., 2007). The high concentrations
detected indicate high consumption of the parent compound in
the studied countries. Moreover, McArdell et al. (2003) pointed out
that variations at the concentrations in the influents between differ-
ent WWTPs in the same country can be attributed to the size of
the WWTP and the type and/or the load of wastewaters that receives
(municipal or hospital wastewaters).
In the effluents the concentrations exhibited lower values up
to 838 ng/L. As an exception, Hirsch et al. (1999) detected very high
ERY-H2O concentrations at WWTP effluents in Germany with a maxi-
mum value of 6000 ng/L. The removal efficiency in WWTPs also varies
and according to McArdell et al. (2003) ERY-H2O and macrolide antibi-
otics in general, cannot be eliminated completely in WWTPs (Fig. 2).
Removal rates up to 19% and between 49 and 80% have been determined
in WWTPs in Hong-Kong (Gulkowska et al., 2008) and Wisconsin, USA
(Karthikeyan and Meyer, 2006). The secondary treatment with longer
hydraulic retention time of the sewage appeared more effective in
reducing ERY-H2O concentrations compared to the primary treatment.
911
Effluents
Occur. Min Max Mean Ref.
Freq.% (ng/L)
81.3 50b 170 118 Letzel et al. (2010)
However, the reason(s) for the difference in removal efficiencies be-
tween WWTPs with common treatment remains largely unknown,
but could be due to the differences in the daily loading of antibiotics
into and the treatment capacity of the individual WWTPs
(Gulkowska et al., 2008).
Moreover, Gulkowska et al. (2008) also observed that concentra-
tions of ERY-H2O in effluents of WWTPs in Hong-Kong were, in some
cases, greater than in influents. Possible explanations for these observa-
tions include (i) deconjugation of conjugated metabolites during the
treatment process (Miao et al., 2002); (ii) an underestimation of the ac-
tual amount due to particulate matter with adsorbed ERY-H2O being
filtered out during sample preparation; and (iii) a change in the adsorp-
tion behavior of the ERY-H2O to particles during treatment processes,
influencing the ratio between influent/effluent water (Lindberg et al.,
2005; Gulkowska et al., 2008) The same observation was reported by
Xu et al. (2007) who attributed this phenomenon to the fact that partic-
ulate matter with sizes greater 0.45 mm was not included leading to an
underestimation of the total content entering the WWTPs. Moreover, a
signal suppression of the MS/MS detector in raw effluent samples due to
high concentrations of organic matter could also be responsible for the
lower concentrations in WWTP influents (Xu et al., 2007).
High concentrations of this TP were also reported in hospital and
drug manufacturing effluents reaching the concentration of a few μg/L
(6110 and 7840 ng/L, respectively) (Lin and Tsai, 2009), suggesting
that this TP can be used as sentinel marker to detect the infiltration of
human used medicines in the surface water systems.
However, ERY-H2O was not the only TP detected for ERY. Gómez-
Ramos et al. (2011) have identified a hydrolysis product of ERY
(Pieper et al., 2010), erythralosamine, in wastewater effluents.
Moreover, Perez-Parada et al. (2011) identified four hydrolysis
products of amoxicillin (AMX) antibiotic, two diastereomers (5S and
5R) of amoxilloic acid ((5S)-AMXO and (5R)-AMXO, respectively) and
two diastereomers (5R and 5S) of AMX diketopiperazine-2′,5′ ((5R)-
AMX-DKP and (5S)-AMX-DKP, respectively). All four TPs of AMX were
detected in both influents and effluents of a WWTP in Spain thus indi-
cating that complete removal of these target compounds after treat-
ment in the WWTP was not possible (Perez-Parada et al., 2011). The
presence of (5R)-amoxicillin diketopiperazine-2′,5′ in Spanish WWTP
effluents was also reported by Gómez-Ramos et al. (2011). However,
erythralosamine and the four TPs of AMX were only identified in waste-
waters but they were not quantified.
Moreover, the hydroxylated derivative of the antibiotic metronida-
zole (METR-OH) was identified by Santos et al. (2013) in hospital efflu-
ents and in WWTP influents and effluents in Portugal. METR-OH is one
of the major metabolites of METR and is biologically active. In hospital
effluents this TP reaches very high concentrations such as 11 μg/L. On
the contrary, the concentrations in WWTP influents ranged at lower
levels (nd–158 ng/L).
Finally, Kosma et al. (submitted for publication) identified two TPs of
the antibiotic trimethoprim in the wastewater effluents in Greece. The
one TP is a keto-derivative identified as (2,4-diaminopyrimidin-5-yl)
(3,4,5-trimethoxyphenyl)methanone and the other is assigned to
912 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926

Table 3
Occurrence of most frequently TPs of β-blockers, statins, antidiabetics, antivirals, estrogens and PCPs in influents and effluents of WWTPs.

Influents Effluents

TPs Occur. Min Max Mean Ref. Occur. Min Max Mean Ref.

(Parent compound/ Freq.% (ng/L) Freq.% (ng/L)

(excretion unchangeda (%))

β-blockers
MTPAb (metoprolol/(10–30)) – 119 298 – Rubirola et al. (2014) – 1107 2506 – Rubirola et al. (2014)
a-HMTPb – b26 36.1 – Rubirola et al. (2014) – n.d n.d – Rubirola et al. (2014)

Statins (antilipidemics)
p-OH-ATVb (atorvastatin/(b2)) 100 n.d 280 280 Vanderford and Snyder (2006) 100 n.d – b1.0 Vanderford and Snyder (2006)
o-OH-ATVb 100 n.d 196 196 Vanderford and Snyder (2006) 100 n.d – b1.0 Vanderford and Snyder (2006)
SMV-OH acidb (simvastatin/(little)) 100 n.d 10 10 Vanderford and Snyder (2006) 100 n.d – b0.5 Vanderford and Snyder (2006)

Antidiabetics
GUAb (metformin/(high)) – n.d 400 400 Trautwein and Kümmerer – n.d – 1860 Trautwein and Kümmerer
(2011) (2011)
67 n.d 19.6 58.9 Kosma et al. 78 n.d 28.3 627 Kosma et al., submitted for
(submitted for publication) publication

Anesthetics
NKb,c (ketamine/(90)) – 1.1 330 – Lin et al. (2014)
– 110 – Lin et al. (2014)

Anticonvulsants
2-N-gluc-LMTGb (lamotrigine/(b10)) – b100 455 – Writer et al. (2013)

Antivirals
OSL-CARb (oseltavir/(90)) – n.d 42.7 42.7 Prasse et al. (2010) – n.d – 17.3 Prasse et al. (2010)
– n.d 29.4 29.4 Prasse et al. (2010) – n.d – 12.2 Prasse et al. (2010)

Stimulants
COTb (nicotine/(−)) 100 780 2880 880 Buerge et al. (2008) 100 10 112 18 Buerge et al. (2008)
– 4 27 15.4 Martínez Bueno – n.d b100 Rosal et al. (2010)
92.9 1.2 42.3 5.1 Huerta-Fontela et al. (2008) – 0.3 10 3.4 Martínez Bueno
35.7 0.8 17.9 2.4 Huerta-Fontela et al. (2008)
– n.d 3095 – Ghoshdastidar et al. (2014)
3-OH-COTb 100 1070 9530 2100 Buerge et al. (2008) 100 60 590 160 Buerge et al. (2008)
N-F-NOR-NICOTb 100 16 149 18 Buerge et al. (2008) 100 15 70 20 Buerge et al. (2008)
PARb (caffeine/(b1)) – n.d 18,214 – Ghoshdastidar et al. (2014)

Estrogens
4-OHE1b (E1/(−)) – n.d 14 – Xiao et al. (2001) – n.d – 27 Gentili et al. (2002)
– 13 15 – Jiang et al. (2005)
– 64 82 – Schlusener (2005)
16α-OHE1b – 1 5 – Ternes et al. (2002)
– 36 71.7 – Jiang et al. (2005)
– 2.4 14 – Schlusener (2005)
E1-3Sb – n.d 0 34.1 Reddy et al. (2005) – – – 0.29 Reddy et al. (2005)
E1-3Gb – n.d 0 0.4 Reddy et al. (2005)
2-OHE2b (E2/(−)) – 6.3 6.3 – Xiao et al. (2001)
– 14 14 – Xiao et al. (2001)
– n.d 15 15 Gentili et al. (2002)
E2-3Sb – n.d 0 3.2 Reddy et al. (2005)
E2-3Gb – n.d 0 0.3 Reddy et al. (2005)

Personal care products

ClMePc (paraben/(−)) 100 12 61 40 González-Mariño et al. (2011) 18 n.d. 6.9 – González-Mariño et al. (2011)
Cl2MePc 100 8 90 46 González-Mariño et al. (2011) 45 n.d. 12 2.6 González-Mariño et al. (2011)
Me-TCSb (Triclosan) – 4 0 – Lindstrom et al. (2002) – b2–11 – – Lindstrom et al. (2002)
4-Cl-TCSc – 2 17 – Buth et al. (2011) – n.d 4 – Buth et al. (2011)
6-Cl-TCSc – 3 98 – Buth et al. (2011) – n.d 9 – Buth et al. (2011)
4,6-Cl-TCSc – 8 29 – Buth et al. (2011) – n.d 22 – Buth et al. (2011)
Dioxinc – 20 8900 – Mezcua et al. (2004) – 4 400 – Mezcua et al. (2004)
a
Petrie et al. (2014).
b
Biotic transformation.
c
Abiotic transformation.

2,4-diaminopyrimidine-5-carbaldehyde. These two TPs were not quan-
tified in WWTP effluents and they were previously identified as photol-
ysis products of trimethoprim (Kormos et al., 2011).
5.2. Lipid regulating agents
Lipid regulators are among the most widespread pharmaceuticals in
wastewaters. However, data concerning the TPs of this drug group ex-
ists only for clofibrate, etofibrate and fenofibrate.
Clofibrate and etofibrate are hydrolysed within the body to yield
the metabolite clofibric acid. Clofibric acid (CLF acid) was frequently
detected in municipal, industrial and hospital wastewaters with
variable concentrations that even reach the μg/L level, like in Portugal
where Salgado et al. (2011) detected concentrations that reached
41.4 μg/L. Discrepancies between countries were also observed depend-
ing on the prescription rates of the parent compounds. For example, CLF
acid was not detected in the Norwegian samples since its parent com-
pounds clofibrate and etofibrate are not prescribed at all in Norway
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926 913

Table 4
Occurrence of most frequently TPs of illicit drugs in influents and effluents of WWTPs.

Influents Effluents

TPs Occur. Min Max Mean Ref. Occur. Min Max Mean Ref.
(Parent compound/
Freq. (%) (ng/L) Freq. (%) (ng/L)
(excretion unchanged⁎ (%))

THC-COOHa – n.d 62.7 62.7 Castiglioni et al. (2006) – n.d – b0.94 Castiglioni et al. (2006)
(tetrahydrocann-abinol/(little)) – n.d 91.2 91.2 Castiglioni et al. (2006) – n.d – 7.2 Castiglioni et al. (2006)
– n.d 96.2 Boleda et al. (2007) 9 b7 22 Bijlsma et al. (2012)
100 21 128 60 Terzic et al. (2010) 36 5.4 72.8 7.9 Postigo et al. (2010)
88 33 489 – Bijlsma et al. (2012)
36 10.6 21.7 18 Postigo et al. (2010)
OH-THCa 71 2.3 90.9 7.6 Postigo et al. (2010) 7 n.d 0.4 0.4 Postigo et al. (2010)
BEa (cocaine/(1–9)) 100 287 2624 612–893 Metcalfe et al. (2010) 100 62 775 90–452 Metcalfe et al. (2010)
– n.d 547.4 547.4 Castiglioni et al. (2006) – n.d – 100.4 Castiglioni et al. (2006)
– n.d 1132.1 1132.1 Castiglioni et al. (2006) – n.d – b0.92 Castiglioni et al. (2006)
87.5 n.d 2307 810 Huerta-Fontela et al. (2007) 68.8 n.d 928 216 Huerta-Fontela et al.
(2007)
– n.d 290 290 Bones et al. (2007) – n.d. 31 – Bones et al. (2007)
100 89 325 186 Terzic et al. (2010) 75 10 351 – Bijlsma et al. (2012)
– n.d 337 149–337 Castiglioni et al. (2011) – 135 4003 – Gilart et al. (2014)
– n.d 1553 1553 Castiglioni et al. (2011)
100 260 3701 – Bijlsma et al. (2012)
– 334 1294 – Gilart et al. (2014)
NBa – 8 n.d – Castiglioni et al. (2006) – n.d – b0.56 Castiglioni et al. (2006)
– n.d 11 5.3–11 Castiglioni et al. (2011) – n.d – 7.5 Castiglioni et al. (2006)
– n.d 51 51 Castiglioni et al. (2011)
– n.d 18.8 18.8 Castiglioni et al. (2006)
NCOa – n.d 4.3 4.3 Castiglioni et al. (2006) – n.d – 0.7 Castiglioni et al. (2006)
– n.d 13.7 13.7 Castiglioni et al. (2006) – n.d – b0.67 Castiglioni et al. (2006)
– n.d 0.9–3.5 0.9–3.5 Castiglioni et al. (2011)
– n.d 28 28 Castiglioni et al. (2011)
CETa – n.d 0.4–5.1 0.4–5.1 Castiglioni et al. (2011) – n.d – 0.2 Castiglioni et al. (2006)
– n.d 23 23 Castiglioni et al. (2011) – n.d – b0.66 Castiglioni et al. (2006)
– n.d 5.9 5.9 Castiglioni et al. (2006)
– n.d 11.5 11.5 Castiglioni et al. (2006)
ECGa – n.d b7.2 b7.2 Castiglioni et al. (2011)
– n.d 199 199 Castiglioni et al. (2011)
EMEa – n.d 58–119 58–119 Castiglioni et al. (2011)
– n.d 346 346 Castiglioni et al. (2011)
AECa – n.d b2.2 b2.2 Castiglioni et al. (2011)
– n.d 35 35 Castiglioni et al. (2011)
AMEa – n.d b7.5 b7.5 Castiglioni et al. (2011)
– n.d b7.5 b7.5 Castiglioni et al. (2011)
6-AMa (heroin/(little)) – n.d 10.4 10.4 Castiglioni et al. (2006) – n.d – b3.08 Castiglioni et al. (2006)
100 3.3 28 12 Terzic et al. (2010) – n.d – b3.08 Castiglioni et al. (2006)
– n.d 11.8 11.8 Castiglioni et al. (2006) – n.d b10–20 Gilart et al. (2014)
– n.d b3.1 b3.1 Boleda et al. (2007)
– n.d 715 Gilart et al. (2014)
NOR-MORa – n.d b25 b25 Boleda et al. (2007)
MOR-3â-D-gluca – n.d 2.5 2.5 Castiglioni et al. (2006) – n.d – b0.48 Castiglioni et al. (2006)
– n.d 18.1 18.1 Castiglioni et al. (2006) – n.d – b0.48 Castiglioni et al. (2006)
EDDPa (methadone/(28)) – n.d 91.3 91.3 Castiglioni et al. (2006) – n.d – 72.1 Castiglioni et al. (2006)
– n.d n.d n.d. Bones et al. (2007) – 9 206 Bones et al. (2007)
100 71 156 128 Terzic et al. (2010) – n.d 22.6 Castiglioni et al. (2006)
– n.d 19.8 19.8 Castiglioni et al. (2006) – b40–60 97 – Gilart et al. (2014)
– 4.5 41.3 Boleda et al. (2007)
– n.d b40–60 b40–60 Gilart et al. (2014)
DICODa (hydrocodone/(12)) 54.5 n.d 330 100 Hummel et al. (2006)
– n.d b10–20 b10–20 Gilart et al. (2014)
NOR-CODa (codeine/(64–70)) – n.d 5 – Boleda et al. (2007)
DIC 83.3 n.d 140 60 Hummel et al. (2006)
– 43 91 – Gilart et al. (2014)
a
COD – 7.3 17 – Wick et al. (2011) – 22 49 – Wick et al. (2011)
14-OH-CODa – n.d b5 b5 Wick et al. (2011) – 2.5 8.4 – Wick et al. (2011)
– n.d b5 b5 Wick et al. (2011) – b2.5 4.3 – Wick et al. (2011)
Neopinea – n.d 7.9 Wick et al. (2011) – 4.7 18 – Wick et al. (2011)
Isoneopine – n.d b5 b5 Wick et al. (2011) – 5 24 – Wick et al. (2011)
NOR-LSDa 7 n.d 4.3 4.3 Postigo et al. (2010) 14 n.d 0.4 0.4 Postigo et al. (2010)
(lysergic acid diethylamide/(b1))
⁎ Petrie et al. (2014).
a
Biotic transformation.

(Weigel et al., 2004). It is also known to be present in rivers and ground No definitive conclusion could be reached on the removal rates of
or tap water (Heberer and Stan, 1997; Lapworth et al., 2012; Leung CLF acid (Fig. 2) warranting further investigation to confirm the results,
et al., 2013). as the removal efficiencies depend on the specific treatment processes
914 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
Fig. 2. Number of studies performed in different countries for the determination of TPs of PPCPs and illicit drugs in wastewaters (data from Tables 1–4 (see also Tables S2–S5)).
adopted in each WWTP and the operational regime of the concerned
WWTPs. Consequently, Ternes (1998) demonstrated a removal of 15%
during trickling filter treatment, 34% during activated sludge treatment
and 51% in an activated sludge system using ferric chloride. Moreover,
Stamatis and Konstantinou (2013) exhibited a 64% removal for CLF
acid after the secondary treatment and 74% after tertiary treatment
with chlorine and sand filtration. Behera et al. (2011) reported 76% re-
moval after the secondary treatment in S. Korea while Salgado et al.
(2012) exhibited very high removal (N75%) after biological treatment
in Portugal and very low removal by adsorption on activated sludge
since only negligible amounts of CLF acid remained undegraded (after
biological removal) and could be found adsorbed to the sludge. Finally,
Roberts and Thomas (2006) demonstrated a 91% removal after tertiary
treatment with chlorine disinfection in UK.
Fenofibric acid (FEN acid), the TP of fenofibrate was detected in
Spain by Rosal et al. (2010) and by Bueno Martínez et al. (2007). This
TP is an active human metabolite and its concentrations in WWTPs
ranged up to 349 ng/L. Rosal et al. (2010) also indicated that FEN acid
was detected in every sample although its precursor was only detected
in one sample and among other compounds had very low removal effi-
ciency after the secondary treatment (1.3%).
5.3. Analgesics
Eleven different TPs of the broadly consumed non-steroid anti-
inflammatory drugs (NSAIDs) are detected in wastewaters. One TP of
acetylsalicylic acid (salicylic acid, SA), three TPs of ibuprofen (1-
hydroxy-ibuprofen (1′-OH-IBU), 2-hydroxy-ibuprofen (2′-OH-IBU),
carboxy-ibuprofen (CX-IBU)), three TPs of diclofenac (5-hydroxy-
diclofenac (5′-OH-DCF), 4-hydroxy-diclofenac (4′-OH-DCF), 4-hydroxy-
diclofenac-dehydrate (4′-OH-DCF-H2O)), one TP of acelofenac (4 -
hydroxyacelofenac, 4′-OH-ACF), two TPs of dimethylaminophenazone
(1-acetyl-1-methyl-2-dimethyloxamoyl-2-phenylhydrazide (AMDOPH)
and 1-acetyl-1-methyl-2-phenylhydrazide (AMPH)) and one TP of
phenazone 1,5-dimethyl-1,2-dehydro-3-pyrazolone (DP) were detected
in WWTP influents and effluents.
Acetylsalicylic acid is rapidly deacetylated in the body to form the
active metabolite SA. SA presented in most cases very high frequency
of detection, almost 100%, and was consistently detected in all samples
(WWTP influents, effluents and in surface water samples collected near
WWTP effluents) (Verenitch et al., 2006). The concentration of SA in the
influents ranged up to 89.1 μg/L (Kosma et al., 2010). In the effluents the
concentrations ranged at lower levels ranging up to 6825 ng/L (Kosma
et al., 2014). However, Cruz-Morato et al. (2013) observed a 46%
increase in the concentration of SA after treatment due to possible
deconjugation of glucuronide metabolites during biological treatment.
On the other hand, Lee et al. (2005) have found very low concentrations
of SA in the effluents of the WWTPs although it was one of the most
abundant acidic pharmaceuticals in the influents. They attributed this
finding to the instability of the compound in sewage samples (Lee
et al., 2003), thus presenting a removal of more than 90%. Stamatis
and Konstantinou (2013) and Kosma et al. (2014) exhibited also high
removal efficiencies of 85% and 70–100% respectively, after the second-
ary treatment thus being in agreement with Henschel et al. (1997) who
demonstrated that SA presents high biodegradability with very high
elimination rate after the secondary treatment. Moreover, Stamatis
and Konstantinou (2013) demonstrated a total removal rate of 90%
after tertiary treatment that was attributed mostly to chlorination
since sand filtration proved inefficient due to its ionized group or to its
low hydrophobicity.
The NSAID ibuprofen is among the most widely used pharmaceuti-
cals in the world. After administration, 15% is excreted as the parent
compound while 26% is excreted as OH-IBU and 43% as CX-IBU, includ-
ing conjugates and carboxy-hydratropic acid (2-phenylpropanoic acid)
in minor amounts (von Bruchhausen et al., 1994). OH-IBU was identi-
fied in WWTP influents and effluents at concentrations reaching
6840 ng/L and 1130 ng/L, respectively; while CX-IBU was detected at
higher concentration levels reaching 38.4 μg/L in the influents and
10.6 μg/L in the effluents (Table 1 & Tables S2 & S3). Under environmen-
tal conditions, these compounds have different transformation kinetics.
No significant transformation along the sewer system and primary
clarification was observed (Stumpf et al., 1998; Weigel et al., 2004).
Zwiener et al. (2002) reported that OH-IBU was formed under aerobic
conditions in activated sludge while carboxy-hydratropic acid was
formed under anaerobic conditions. In both cases, these TPs did not
account for more than 10% of the initial IBU concentration, suggesting
that the major amounts in sewage come from human excretion.
However, Stumpf et al. (1998) reported that major changes occur on
these TPs during biological treatment where CX-IBU is almost quantita-
tively eliminated while OH-IBU is hardly affected and consequently it is
the dominant TP in WWTP effluents. On the contrary, Bendz et al.
(2005) reported high removal efficiencies not only for CX-IBU (96%)
but also for OH-IBU (95%). This contradiction was attributed to the spe-
cific operational parameters of the activated sludge process which were
found to be crucial for the removal of OH-IBU. The same observation
was also previously demonstrated by Buser et al. (1999) who reported
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
that both metabolites of IBU are easy to be eliminated during sew-
age treatment. Generally, in most environmental samples OH-IBU
appears to be the dominant compound while on the contrary Weigel
et al. (2004) reported that CX-IBU is the dominant compound in
seawater samples thus indicating that the fate of these substances
strongly depends on environmental conditions (temperature, salin-
ity, pH, biological activity). The investigation of parent compound/
metabolite concentration patterns will be a valuable tool in the further
assessment of the environmental fate of ibuprofen and pharmaceuticals
in general.
Moreover, Batt et al. (2008) reported the presence of 2′-OH-IBU
in WWTP effluents in USA. 2′-OH-IBU is the main hydroxyl derivative
that is produced via metabolism, while 1′-OH-IBU and 3′-OH-IBU
were also found in urine but at very small concentrations (Kepp et al.,
1997); however, there is scarce data for their presence in wastewaters.
This is in agreement with Pérez and Barceló (2007) who pointed out
that the three isomers of OH-IBU could not be differentiated unequivo-
cally based on their mass spectra alone, but the most likely metabolite
was believed to be 2′-OH-IBU. Moreover, Ferrando-Climent et al.
(2012), identified 2′-OH-IBU and 1′-OH-IBU in WWTP influents and ef-
fluents. In accordance with the aforementioned study, 1′-OH-IBU
ranged at much lower values in influents and effluents (2.55–5 μg/L
and 0.11–1.41 μg/L respectively) than 2′-OH-IBU (1.21–93.65 μg/L and
0.39–5.87 μg/L respectively), which appeared to be the dominant me-
tabolite. It is worth mentioning that the hydroxylated derivatives of
IBU appear to be more toxic than the parent compound increasing the
inhibition percentage of bioluminescence from V. fischeri as indicated
by Méndez-Arriaga et al. (2008).
DCF is another widely prescribed NSAID and it is mainly metabolized
in humans to its hydroxylated or methoxylated derivatives and further
conjugated, mostly to glucuronides (Sawchuk et al., 1995). A total of 6%
(parent drug including its conjugates) of a prescribed dose is excreted in
the urine. However, excretion of the main metabolite 4′-OH-DCF (4′-
OHD, {2-[(2,6-dichloro-4-hydroxyphenyl)amino]phenyl}acetic acid)
and its conjugates in human urine represents 16% of the dose. 3′-OH-
DCF, 5′-OH-DCF ({2-[(2,6-dichlorophenyl)amino]-5-hydroxyphenyl}
acetic acid), and 3′-hydroxy-4′-methoxy DCF together with their conju-
gates represent 2.0%, 6.1% and 0.01%, respectively. Moreover, 4′,5-
dihydroxy DCF and its conjugates were found in human urine at about
9.4% of the dose (Schneider and Degen, 1981). The 4′-hydroxy DCF de-
hydrate (4′-OHDD, 1-(2,6-dichloro-4-hydroxyphenyl)-1,3-dihydro-
2H-indol-2-one) was also identified as a minor human metabolite of
DCF (Stülten et al., 2008).
Stülten et al. (2008) detected 4′-OH-DCF, 5′-OH-DCF and 4′-OHDD
in WWTP effluents in Germany at concentrations up to 1600 ng/L,
860 ng/L and 660 ng/L, respectively. The author also observed seasonal
variations with higher concentrations from late April and onwards as
the weather was hot and dry. However, great discrepancies in the ratios
of DCF and its metabolites between different WWTPs were observed.
This was attributed to the type of DCF dispensing (orally or by dermal
application). Compared to urinary excretion after oral intake, only 6%
of DCF is absorbed after its application to the skin, and the rest is washed
off (Schrey and Wilhelm, 1999) unchained directly to the wastewaters.
Consequently, higher DCF concentrations are observed compared to its
metabolites, when dermal applications of the parent compound take
place mostly, while on the contrary, when oral intake is the main appli-
cation, smaller differences between diclofenac and the metabolites
are observed. Likewise the proportions between the metabolites
concentrations also differed. These variations might originate from the
populations of microorganisms that preferentially degraded one of the
compounds or led to the different amounts after cleavage of glucuro-
nide conjugates in the related sewage systems (Halling-Sørensen
et al., 1998). 4′-OH-DCF was detected at higher concentration ratios
(9–44% of DCF concentrations) and was considered as the main metab-
olite detected in WWTP effluents (Stülten et al., 2008). Moreover, Pérez
and Barceló (2008) also detected 4′-OH-DCF in WWTP influents and
915
effluents from CAS (continuous activated sludge) and MBR (membrane
bioreactor) systems, exhibiting 26% and 56% removal, respectively.
Pérez and Barceló (2008) identified also a TP of ACF, 4′-OH-ACF [2-
(2′,6′-dichloro-4′-hydroxyl-phenylamino)phenyl] acetoxyacetic acid
which is the major human metabolite of ACF after oral consumption
(Bort et al., 1996b). The removal efficiency of the parent compound in
the tested WWTPs was 50 to 58% and was higher than the removal
efficiency of DCF. This substantially higher removal rate of ACF com-
pared to DCF (17%) is believed to be due to esterase-mediated enzymat-
ic hydrolysis of the ester moiety of ACF, giving rise to the corresponding
acid which is the DCF. This process, which is also a relevant in vivo bio-
transformation pathway, involving the conversion of ACF into DCF and
4′-OH-ACF into 4′-OH-DCF (Bort et al., 1996a), can be attributed to
esterase activity in the activated sludge, which contains structurally di-
verse esterase enzymes with broad, and partially overlapping, substrate
specificity (Boczar et al., 2001). Concentrations of 4′-OH-ACF were
generally higher than the parent compound reaching 82 ng/L in the in-
fluents and 20 ng/L and 1.8 ng/L in the effluents after CAS and MBR
treatment, respectively (Pérez and Barceló, 2008).
Taken together, the study on the occurrence and fate of ACF and DCF
in the WWTP illustrates that human drug metabolites can (a) be at least
as prominent in untreated sewage as the parent compound and
(b) display a distinct behavior in the biological wastewater treatment
(Pérez and Barceló, 2008).
Zuehlke et al. (2004) detected two oxidation products of
dimethylaminophenazone: 1-acetyl-1-methyl-2-dimethyloxamoyl-2-
phenylhydrazide (AMDOPH) and 1-acetyl-1-methyl-2-phenylhydrazide
(AMPH). AMDOPH was also identified as a product of photochemical de-
composition of dimethylaminophenazone (Marciniec, 1984). The con-
centrations of these two TPs were similar in the influents and effluents
of the WWTP in Germany reaching 110 ng/L in both cases, thus present-
ing negligible removal after the secondary treatment. The authors also
detected 1,5-dimethyl-1,2-dehydro-3-pyrazolone (DP) which was con-
sidered as a metabolite of phenazone, another antipyretic drug, and
found slightly higher concentrations in the effluents compared to the
wastewater influents of the WWTP in Germany (Zuehlke et al., 2007).
Besides NSAIDs, TPs of analgesic drugs such as metamizol (also
known as dipyrone) were also detected in wastewaters. The occurrence
of dipyrone residues in WWTP effluents has been less frequently
assessed but Feldmann et al. (2008) have reported concentrations up
to 7 μg/L in influents and effluents of a WWTP in Berlin. After human
metabolism of metamizole, 4-methylaminoantipyrine (4-MAA) is pro-
duced which is further metabolized to give-formyl-4-aminoantipyrine
(4-FAA) and 4-AA (4-aminoantipyrine) which is then acetylated to 4-
acetyl-amino-antipyrine (4-AAA) (Carretero et al., 1995; Schmidt
and Brockmeyer, 2002). 1,5-Dimethyl-1,2-dehydro-3-pyrazolone (DP)
and 4-(2-methylethyl)-1,5-dimethyl-1,2-dehydro-3-pyrazolone (PDP)
were identified as microbiological metabolites and could also appear
in the aquatic environment (Zuehlke et al., 2004).
Monitoring studies confirmed the presence of the metabolites
of dipyrone and its active product 4-MAA such as 4-AAA, 4-FAA, 4-
DAA (4-dimethylaminoantipyrine), 4-AA and (PDP) in WWTP influents
and effluents (Bueno Martínez et al., 2007; Rosal et al., 2010; Zuehlke
et al., 2004). According to the data shown in Table 1 (see also
Tables S2 & S4), the concentrations of these metabolites in influents
and effluents of WWTPs were elevated and in all cases were at μg/L
levels. Higher concentrations were observed for 4-AAA with values up
to 22.2 μg/L and 6.75 μg/L and for 4-FAA with values up to 71 μg/L
and 27.4 μg/L in the influents and effluents, respectively. Removal
efficiencies after the secondary treatment were calculated by Rosal
et al. (2010) and ranged for all dipyrone metabolites between 46%
(for 4-AAA) and 68% (for 4-FAA). Gómez et al. (2008) demonstrated
that 4-MAA, 4-AAA and 4-FAA do not exhibit significant acute
toxicity to Daphnia magna. The inhibition values on the motility of the
Daphnis were within 0–20% and 5–40%, after 24 and 48 h culture,
respectively.
916 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
5.4. Antidepressants
Because of their transformation rates and their pharmacological
activity many metabolites of the antidepressants have been determined
in wastewaters (Lajeunesse et al., 2008). However, it should be recog-
nized that the metabolism of antidepressants may vary widely between
individuals, and by age and sex (Ronfeld et al., 1997; Baumann, 1998),
so the distribution of excretion products in wastewater will vary with
the demographics of the population served by the WWTP (Metcalfe
et al., 2010). Moreover, the degree of metabolism varies from com-
pound to compound, but for most SSRIs the rate is relatively high.
Desmethyl (e.g. desmethyl sertraline, N-desmethyl venlafaxine, O-
desmethyl venlafaxine, desmethyl citalopram, didesmethyl citalopram,
desmethyl diltiazem, nortriptyline) or hydroxyl (e.g. 10-OH-ami-
triptyline, 3-OH-diazepam, OH-bupropion, erythro-threo hydrobu-
propion) metabolites are among the most frequently detected TPs of
antidepressants in WWTP influents or effluents. According to the data
shown in Table 2 higher concentrations in wastewaters were measured
for O-desmethyl-venlafaxine (up to 5500 ng/L), OH-bupropion (up to
2000 ng/L) and erythro-threo hydrobupropion (up to 5700 ng/L).
Desmethyl-citalopram, on the other hand, was consistently identified
but ranged at lower values (up to 426.7 ng/L) (Writer et al., 2013).
The high concentrations of O-desmethyl venlafaxine and eryhro-
threo-hydrobupropion in wastewaters are explained by the fact that
these Phase II metabolites are the dominant metabolites in human
urine, and they are found at much higher concentrations than the
parent compounds in raw wastewater. Moreover, Writer et al. (2013)
calculated the metabolite: parent compound ratio for O-desmethyl
venlafaxine–venlafaxine and threo-hydrobupropion–bupropion in
wastewater effluents and they have found lower values than those
calculated in urine. This phenomenon can be possibly explained as fol-
lows: 1) the metabolites are either more labile relative to the parent
compounds and being degraded by wastewater treatment processes,
2) the parent compounds are being reformed from the metabolites,
and/or 3) parent compounds are present at higher concentrations
due to the disposal of unused medications. For all other antidepressants
and their TPs, the conjugated metabolites predominate in urine and
the unconjugated (free) forms of these compounds are present in low
proportions in urine thus explaining the lower detected concentrations
in wastewaters (Schroeder, 1983; Benfield et al., 1986; Kaye et al., 1989;
Warrington et al., 1992). Furthermore, norfluoxetine and desmethylser-
traline were also very abundant exhibiting in some cases 100% frequen-
cy of detection (Vanderford and Snyder, 2006; Vasskog et al., 2008).
Generally, the concentrations of TPs of antidepressants are lower in
the WWTP outflows than in the WWTP inflows. However, Vasskog
et al. (2008) showed that WWTPs with the secondary treatment did
not achieve better removal of these metabolites than WWTPs with the
primary treatment. This indicates that at least a part of the removed
amount is bound to particles, and is eliminated in the filtration process
in both cases. This is in agreement with the research of Writer et al.
(2013) who also pointed out that lower concentrations of antidepres-
sants and their metabolites in the effluents of trickling filter treatment
as compared to activated sludge treatment. They also found no linear
correlation (r2 = 0.2, n = 17) between the hydraulic retention time of
each wastewater treatment plant and the summed concentration of
targeted neuro-active compounds and their metabolites. Consequently,
conventional wastewater treatment does not completely remove anti-
depressant pharmaceuticals and their metabolites and WWTPs are a
point source of these compounds to the environment (Vasskog et al.,
2008).
5.5. Psychoactive drugs
Studies in Europe and North America have shown that carbamaze-
pine (5H-dibenzo[b,f]azepine-5-carboxamide) (CBZ) is one of the
most frequently detected pharmaceuticals in WWTP effluents and in
river water (Ternes, 1998; Ollers et al., 2001; Heberer, 2002; Metcalfe
et al., 2003). CBZ is predominantly metabolized in the liver, and at
least 30 different metabolites have been identified (Lertratanangkoon
and Horning, 1982). Three key metabolic pathways have been reported
(Breton et al., 2005). The main pathway for CBZ is the oxidation leading
to the formation of CBZ-10,11-epoxide (CBZ-Ep), a pharmacologically
active compound with anticonvulsant properties. Subsequently, hydra-
tion takes place and CBZ-Ep is further metabolized into 10,11-dihydro-
10,11-trans-dihydroxy-carbamazepine (DiOH-CBZ) which in turn leads
to the formation of 9-hydroxymethyl-10-carbamoylacridan (9ac). The
DiOH-CBZ metabolite is not pharmaceutically active. The second meta-
bolic route involves the production of hydroxylated compounds 2-OH-
CBZ and 3-OH-CBZ while 4-OH-CBZ plays a minor role. The third
minor metabolic pathway leads to iminostilbene (IM) formation.
Furthermore, within leukocytes, CBZ and IM are metabolized into
oxidative products, including acridine (AI) and acridone (AO), which
are known to be genotoxic (Bleeker et al., 1999; Miao et al., 2005;
Leclercq et al., 2009).
Oxcarbazepine (OxCZ), a keto analog of CBZ, which is used as an an-
ticonvulsant, also generates the metabolites DiOH-Cbz and 9ac. Howev-
er, OxCZ is mainly metabolized into 10-hydroxy-10,11-dihydro-
carbamazepine (10-OH-CBZ), its therapeutically active metabolite. Fur-
thermore, Li et al. (2011) also demonstrated the formation of DiOH-CBZ
and acridine as photoproducts of OxCZ. More specifically, the authors
demonstrated that acridine was generated by DiOH-CBZ which
underwent a ring contraction process probably upon direct photolysis.
All the aforementioned TPs of CBZ and OxCZ have been identified in
WWTP influents and effluents (Table 2). The metabolite DiOH-CBZ was
the predominant analyte in the aqueous phase with concentrations
higher than the parent compound, reaching few μg/L in influents and ef-
fluents as well (up to 4000 and 3400 ng/L, respectively). On the other
hand, CBZ-Ep, which is present in human plasma at 3- to 4-fold lower
concentrations than DiOH-CBZ, was found in some cases at concentra-
tions at least 50-fold lower than those of DiOH-CBZ in sewage
(Leclercq et al., 2009). This is consistent with the known instability of
epoxides in aquatic solutions. CBZ-Ep is probably excreted at much
lower concentrations and also transformed into DiOH-CBZ in sewage.
On the contrary, Writer et al. (2013) exhibited that although 10-OH-
CBZ is not a significant human metabolite of CBZ according to pharma-
cokinetic studies (Reith et al., 2000) it was routinely found at concentra-
tions higher than CBZ, and detected more frequently than other CBZ
metabolites. One possible explanation for this conflicting information
is that 10-OH-CBZ is being transformed from OxCZ. However, there
was no correlation between OxCZ and 10-OH-CBZ detection and con-
centration levels. It is also unclear whether this is due to changes in
patient treatment (switching from CBZ to OxCZ) or microbial formation
of 10-OH-Cbz from CBZ and/or other metabolites in the wastewater
treatment processes suggesting that these valuable findings warrant
further investigation (Writer et al., 2013). Moreover, Bahlmann et al.
(2014) also identified 4-OH-CBZ in wastewater influents and effluents
of WWTPs in Germany and Portugal. Although not quantifiable, 4-OH-
CBZ was observable in both influent and effluent wastewaters suggest-
ing a low removal rate for this compound.
Generally no loss of the metabolites of CBZ was observed during
conventional activated sludge treatment. However, in some cases
the concentrations of 2-OH-CBZ and DiOH-CBZ increased significantly
after activated sludge treatment. This phenomenon is attributed to
the fact that CBZ and its metabolites have been found to be partially ex-
creted as glucuronide conjugates (N-glucuronide of CBZ, N-glucuronide
of 10,11-dihydro-10,11-epoxycarbamazepine, O-glucuronide of trans-
10,11-dihydroxy-carbamazepine, and isomeric O-glucuronides of
hydroxy-, dihydroxy-, and hydroxymethoxycarbamazepine have been
identified as urinary metabolites) (Lynn et al., 1978; Lertratanangkoon
and Horning, 1982; Maggs et al., 1997; Schutz et al., 1986). Furthermore,
activated sludge was found to have glucuronidase activity (Ternes et al.,
1999a,b), so consequently, cleavage of the glucuronic acid moiety is
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
possible in WWTPs, thus leading to the release of parent compounds
and metabolites in free form, as suggested by several authors (Ternes,
1998; Vieno et al., 2006). Leclercq et al. (2009) also proposed that it is
likely to occur cleavage in the sewage network especially when the res-
idence time is long and that the cleavage of glucuronide conjugates by
enzymatic process in treatment plants utilizing activated sludge or
trickling filter system is the most probable explanation for the constant
presence of 10-OH-CBZ, DiOH-CBZ and 2-OH-CBZ metabolites in the
effluents.
Moreover, Soufan et al. (2013) demonstrated the formation of CBZ-
Ep and of a hydroxylated derivative of CBZ during chlorination. Conse-
quently, the formation of these TPs may also occur in the WWTP
when chlorination follows the secondary treatment. CBZ-Ep was proved
to be quite stable during chlorination while on the other hand the hy-
droxylated derivative's concentration was reduced during chlorination
time.
Ortiz de García et al. (2013b) estimated the ecotoxicological risks of
several metabolites of CBZ by employing quantitative structure activity
relationship (QSAR). Regarding the chronic toxicity for marine or fresh-
water organisms, the level of concern estimated for Ep-CBZ was similar
compared to the parent compound, while higher levels of concern were
estimated for DiOH-CBZ, 2-OH-CBZ and 3-OH-CBZ. Furthermore, pre-
liminary results suggest a higher acute toxicity of 2-OH-CBZ and 3-
OH-CBZ for V. fischeri compared to the toxicity of CBZ (Kaiser et al.,
2013).
Writer et al. (2013) have identified 2-N-glucuronide-lamotrigine as
a metabolite of lamotrigine another antiepileptic drug. Concentrations
of this TP fluctuated from b100 to 455 ng/L but it was only detected in
one WWTP effluent. Furthermore, ritalinic acid was detected in waste-
waters as the metabolite of the drug methylphenidate (known as rita-
lin) (Letzel et al., 2010). After consumption, methylphenidate is easily
adsorbed and quickly metabolized to 2-phenyl-2-(2-piperidinyl)acetic
acid (ritalinic acid). Ritalinic acid shows none or very low pharmaceuti-
cal activity. About 80% of the methylphenidate dosage is eliminated via
urine as ritalinic acid. Ritalinic acid was frequently detected by Letzel
et al. (2010) in influents and effluents of WWTP in Germany (13 of 16
samples) with concentrations presenting a high variability ranging
from b50 to 270 ng/L and from b50 to 170 ng/L, respectively (see
Table 2). Concerning the removal of ritalinic acid after the secondary
treatment the results obtained fluctuating from no removal to 44.9%
with a mean value of approximately 23%. Letzel et al. (2010) found no
possible explanation for these discrepancies since all sampling series
were taken under similar conditions (e.g. weather, season or behavior
of the sewage treatment plant) and suggest that further investigation
should be conducted in the future.
5.6. β-Blockers
β-Blockers are a highly consumed group of pharmaceuticals used to
treat high blood pressure (hypertension) as well as patients recovering
from heart attacks. Among β-blockers, metoprolol (MTP) is a compound
of high consumption (Dong et al., 2013) and its metabolism in mam-
mals leads to the formation of three main metabolites, metoprolol acid
(MTPA), a-hydroxymetoprolol (a-HMTP) and O-desmethylmetoprolol
(O-DMTP) which account for the 85% of the urinary excretion (Barclay
et al., 2012). MTPA is known to be the major human metabolite of
MTP and it can also be formed from its parent compound as a biodegra-
dation product (Kern et al., 2012). However, Radjenovic et al. (2008)
reported the formation of MTPA as a biodegradation product of another
β-blocker, atenolol.
Rubirola et al. (2014) identified MTPA and a-HMTP in WWTP influ-
ents and effluents in Spain (Table 3). MTPA was detected in the influents
at concentrations up to 298 ng/L, which is a 10 fold higher level than the
concentration of MTP in the same samples. However, it matches the ex-
cretion ratios since 60–65% of MTP is excreted as MTPA after human
metabolism (Kern et al., 2012). On the contrary, almost 10 times higher
917
levels of this TP were identified in the effluents reaching 2506 ng/L.
Rubirola et al. (2014) attributed the higher concentrations of this TP in
the effluents not only to the biodegradation of the parent compound
but also to the to the generation of this compound from other β-
blockers present in the WWTP like atenolol. a-HMTP was detected
only in WWTP influents at concentrations up to 36 ng/L. Its removal
was attributed by the authors to its biodegradation. Finally, O-DMTP
was not detected neither in the influents nor in the effluents of the se-
lected WWTPs. Its absence in the influents was due to its rapid transfor-
mation to MTPA in human metabolism (Barclay et al., 2012) while in the
effluents, although a small proportion of MTP is biodegraded to O-
DMTP, the latter is very fast removed according to batch experiments
conducted by the authors. Toxicity tests with V. fischeri didn't show
any acute toxicity of a-HMTP and MTPA (Rubirola et al., 2014).
6. Statins (antilipidemics)
Statins are a group of drugs used for the reduction of the level of cho-
lesterol in blood by reducing the production of cholesterol in the liver.
The best-selling statin is atorvastatin which by 2003 became the best-
selling pharmaceutical in history (Simons, 2003). However, only one re-
port exists concerning the presence of the metabolites of statins in
wastewaters. Vanderford and Snyder (2006) identified p-hydroxy ator-
vastatin and o-hydroxy-atorvastatin as metabolites of atorvastatin and
simvastatin hydroxyl acid as the metabolite of simvastatin at concentra-
tions of 280 ng/L, 196 ng/L and 10 ng/L respectively, in the influents of a
WWTP. All identified metabolites were also present in the effluents as
well, but in very low concentrations (b1.0 or b0.5 ng/L) thus exhibiting
very good removal in the WWTPs.
6.1. Antidiabetics
Metformin (MTF) is among the top applied drugs that can be used
for treatment of all stages of diabetes disease. MTF is not metabolized
in the human body and its elimination only occurs via the kidneys
(Pentikainen et al., 1979). However, Trautwein and Kümmerer (2011)
have proven that MTF is aerobically biodegraded to guanylurea (GUA),
which is a very recalcitrant TP, being stable against further photo- and
biodegradation. Concentration of GUA in the influents of one WWTP
in Germany was found to be 0.4 mg/L but in the effluents was
1.86 mg/L indicating the biotransformation of MTF to GUA in the treat-
ment plant. Trautwein and Kümmerer (2011) also pointed out that the
concentration of GUA in the influents might stem from the transforma-
tion of MTF by bacteria in the sewer system. Finally, in a comprehensive
study of Kosma et al. (submitted for publication) in 8 WWTPs of Greece,
the systematic occurrence of GUA in the influents and the effluents of all
WWTPs was demonstrated. Concentrations up to 58.9 and 627 ng/L
have been determined in the influents and effluents, respectively.
Concentration of GUA mostly exceeds its parent MTF in the effluents,
presenting negative removal efficiencies.
6.2. Anesthetics
Ketamine is a drug with multiple applications but mainly for starting
and maintaining general anesthesia. Ketamine is not completely metab-
olized in humans and other organisms. Norketamine (NK), which has
one-third to one-half the potency of ketamine, has been reported to
be the major metabolite (Tanaka et al., 2005). Lin et al. (2014) identified
NK in hospital effluents with concentrations up to 330 ng/L and in one
WWTP's effluents up to 110 ng/L. The authors have also reported the
formation of NK during photolysis of ketamine. The presence of NK in
the effluents was largely attributed to its excretion through urine and
to a small portion via phototransformation of ketamine to NK. Toxicity
measurements conducted by the authors using V. fischeri showed that
NK exhibited higher toxicity than ketamine.
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6.3. Antiviral drugs
Besides the extended use of antiviral drugs (Schwabe and Paffrath,
2008) scarce data exists concerning their occurrence in the environ-
ment and fate. Moreover, considerable concern has been raised about
the potential environmental effects of anti-influenza drugs which
undergo seasonal variations, with peak emissions during influenza
epidemics (Singer et al., 2008).
Prasse et al. (2010) have identified the TP of the antiviral drug
oseltamivir (OSL) (marked as Tamiflu), oseltamivir carboxylate (OSL-
CAR) in wastewaters and its release into the environment might cause
serious effects such as the generation of OSL-CAR resistance in wildfowl,
taking into account that birds are competent hosts of influenza viruses
(Olsen et al., 2006). The concentration of OSL-CAR measured by Prasse
et al. (2010) in the influents and the effluents of WWTPs in Germany
was higher than the parent compound but ranged in low levels with
concentrations up to 42.7 ng/L and 17.3 ng/L, respectively. They have
also calculated the ratio of OSL to OSL-CAR OP/OC. In raw wastewater
a mean ratio of 0.3 was calculated, which is attributed to the metabolism
of the parent compound OSL in the human body yielding 70–80% OSL-
CAR. On the contrary, in treated wastewater the ratios were 0.72 ±
0.23 and 0.91 ± 0.20 for two different WWTPs due to the reduced con-
centrations of OSL-CAR and the stability of OSL (Prasse et al., 2010).
6.4. Proton pump inhibitors
Omeprazole (OME) is one the most frequently prescribed and ad-
ministrated proton pump inhibitor drugs in humans (Ortiz de García
et al., 2013a). Only one report exists concerning the occurrence of
OME metabolites in wastewaters. Boix et al. (2014) identified up to 14
metabolites of omeprazole in wastewater influents and effluents.
Eleven of these compounds were identified in all water matrices ana-
lyzed, even at surface water. Unchanged OME was not identified in
any sample. The most abundant metabolite (named by Boix et al.
(2014) as OM10) is a product of sulfoxide reduction and subsequent
hydroxylation and oxidation from parent OME and exhibited 100%
frequency of detection in WWTP influents or effluents in Spain. It is
noteworthy that up to eight metabolites were detected in 90–100% of
effluent wastewater samples. These OME metabolites were also present
in influent wastewater, although some of them at a lower frequency.
This was attributed by the authors to the higher complexity and strong
signal suppression due to matrix effects in the influent that made the
detection of OME metabolites more problematic in comparison with ef-
fluent wastewater. All these metabolites of OME were only identified
and not quantified in wastewater samples.
6.5. Anticancer (cytostatic drugs)
Anticancer drugs are considered nowadays as an important class of
environmental contaminants since 75% of the treated patients with
chemotherapy leave the hospital after treatment and therefore, the
cytostatic compounds can reach the aquatic environment via domestic
wastewater in addition to hospital wastewater and ultimately through
WWTPs. Moreover, they have been reported to show low biodegrad-
ability by conventional wastewater treatments and are thus considered
recalcitrant compounds (Lenz et al., 2007; Kazner et al., 2008). Cytostatic
agents exhibit potent cytotoxic, genotoxic, mutagenic, carcinogenic and
teratogenic effects in several organisms since they have been designed
to disrupt or prevent cellular proliferation, usually by interfering in
DNA synthesis (Santos et al., 2010). Moreover, their metabolites might
have the same mode of action or even higher activity than the parent
compounds. For example, hydroxy-tamoxifen (OH-TMX), a metabolite
of tamoxifen exhibits higher estrogenicity than the parent compound
(Custodio et al., 1994).
However, scarce data exists concerning their occurrence in waste-
waters. Ferrando-Climent et al. (2013) reported the occurrence of
OH-TMX, the main human metabolite of tamoxifen, 4,4-dihydroxy
desmethyltamoxifen (endoxifen), which belongs to the same chemical
and therapeutic family of TMX and is also a secondary human metabo-
lite of tamoxifen and carboxy-phosphamide, a known human metabo-
lite of cyclophosphamide. Ferrando-Climent et al. (2013) have only
identified these compounds in hospital effluents but they did not quan-
tify them. Finally, apart from TMX, 2′,2′-difluorodeoxyuridine, the main
metabolite of gemcitabine, was also detected in hospital wastewater
(Weissbrodt et al., 2009).
6.6. X-ray contrast media
Iodinated X-ray contrast media (ICM) are the most widely used
pharmaceuticals for the imaging of internal organs, blood vessels and
soft tissues during radiological and medical diagnostic procedures.
They are administrated intravascularly at high daily doses (up to
200 g/d), are metabolically stable in the body and are rapidly eliminated
via urine or feces; therefore are frequently detected in wastewater at
elevated concentrations (sometimes N10 μg) (Pérez and Barceló, 2007).
In a comprehensive research on fate and occurrence of ICM
(iopromide, iohexol, iomeprol and iopamidol) carried out by Ternes
and co-workers (Ternes and Hirsch, 2000; Schulz et al., 2008; Kormos
et al., 2009, 2010, 2011), a number of TPs of this class of agents have
been detected in effluents of WWTPs in Germany. It was found that bio-
transformation occurred only in the side chains of the ICMs. In a conven-
tional WWTP a high transformation rate (N80%) was observed for
iohexol, iomeprol, and iopromide, whereas iopamidol was transformed
to a minor extent with a calculated transformation rate of ~35%.
Specifically, 12 TPs of iopromide were detected at least once with a
sum of up to 11.9 μg/L in the effluents of three WWTPs (Schulz et al.,
2008). According to the authors, the higher detectable amounts were
for TPs formed at the initial stages of the biotransformation pathway
of iopromide through WWTP or for compounds which had already
lost their R-hydroxypropionic acid side chain by amide-methylene
bond scission. In addition to 12 iopromide TPs, one iopamidol TP
(TP791) and 10 iomeprol TPs were also detected in effluents with con-
centration levels up to 5.7 μg/L. It is worth to point out that many of the
identified TPs could also be detected in environmental samples (surface
water, groundwater) and even in drinking water (Kormos et al., 2009,
2010, 2011).
Currently, there is limited ecotoxicological information available for
ICM and their TPs, thus, more data and further refinement of the risk
assessment are required to estimate the acute and chronic potential
effects of the parent compounds and their TPs or their mixtures.
6.7. Stimulants
Nicotine is rapidly and extensively metabolized in the human liver
and approximately 70–80% of the nicotine absorbed by a smoker is
transformed to cotinine and excreted in the urine (Bramer and
Kallungal, 2003). Cotinine has been detected in wastewaters at very
high concentrations even reaching the μg/L level, e.g. 42.3 μg/L in the in-
fluents of a WWTP in Spain (Huerta-Fontela et al., 2008) and 3.09 μg/L
in the effluents of WWTPs in Canada (Ghoshdastidar et al., 2014).
Except cotinine, Buerge et al. (2008) have also detected the other
major urinary metabolite of nicotine, 3′-hydroxycotinine, as well as a
further minor tobacco alkaloid, N-formylnornicotine in samples from
Swiss WWTPs.
Concentrations of the two major urinary metabolites of nicotine,
cotinine and 3-hydroxycotinine are generally lower in the effluents
than in the influents thus indicating a good removal of these compounds
in the WWTPs after tertiary treatment that even reach 98% and 93% re-
spectively (Buerge et al., 2008). On the other hand, N-formylnornicotine
exhibits high stability during wastewater treatment. According to
Buerge et al. (2008) the concentrations of N-formylnornicotine in un-
treated wastewaters are 2–3 orders of magnitude lower than the two
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
major urinary metabolites and the typical concentration ratio cotinine/
3′-hydroxycotinine/N-formylnornicotine is ~50:120:1. On the contrary,
in the treated wastewaters (after tertiary treatment), this ratio changes
(~1:8:1) in favor of N-formylnornicotine since the measured concentra-
tions in the effluents are similar to those measured in the influents.
Moreover, incubation experiments conducted by the authors in the lab-
oratory (using untreated wastewater and activated sludge from a mu-
nicipal WWTP) showed no dissipation within 24 h of incubation thus
confirming the persistence of this metabolite in WWTPs.
Due to the widespread consumption of tobacco, cotinine has been
suggested as a potential chemical marker for domestic wastewater con-
tamination of surface waters (Metcalfe et al., 2003; Glassmeyer et al.,
2005; Benotti and Brownawell, 2007; Bradley et al., 2007). Moreover,
it is already used as an indicator of tobacco smoke exposure (Haufroid
and Lison, 1998). However, Buerge et al. (2008) demonstrated also
the suitability of the other nicotine derivatives as hydrophilic, anthropo-
genic markers. Finally, Ghoshdastidar et al. (2014) have reported high
concentrations of paraxanthine (PAR), a metabolite of caffeine, at
WWTPs effluents in Canada reaching 18.2 μg/L.
6.8. Estrogens
The presence of estrogenic compounds in the aquatic environment is
of great concern since these substances can cause endocrine disruption
and interferences in reproduction and development of aquatic organ-
isms (Routledge et al., 1998b; Tyler et al., 1998). Estrone and estriol
are the basic urinary metabolites of estradiol. Although a plethora of
studies exists concerning the occurrence of estrogens in wastewaters,
scarce data is available for the presence of their metabolites. Xiao et al.
(2001) have identified 2-hydroxy estrone (2-OHE1), 4-hydroxy-
estrone (4-OHE1) and 2-hydroxy-estradiol (2-OHE2) in WWTP influ-
ents in UK. All three metabolites ranged at low concentration levels up
to 14 ng/L. Moreover, the occurrence of another metabolite of estrone,
16-a-hydroxy-estrone is reported at concentrations ranging from 1 to
71.7 ng/L in the influents and from 13 to 82 ng/L in the effluents of
WWTPs in Germany and UK (Table 3). Jiang et al. (2005) demonstrated
that concentrations of estrogenic compounds are generally reduced
after activated sludge and sedimentation. However, sand filtration
does not help to remove residual EDCs significantly (Jiang et al., 2005).
Additionally, steroid conjugates were also detected in the wastewa-
ters. Steroid hormones including E1 and E2 are excreted in urine of
humans and animals as the more hydrophilic glucuronides and sulfates
(Shackleton, 1986). Although, these steroid conjugates do not have af-
finity to estrogen receptors evidence suggests that once the conjugated
estrogens reach sewage treatment plants, they undergo chemical or en-
zymatic dissociation in bacterial sludge and reform active estrogens
(Ternes et al., 1999a,b). Consequently, they can be considered as poten-
tial sources of active estrogens due to their dissociation in sewage or as a
result of the input of untreated wastewater directly into surface waters.
β-Estradiol-2-sulfate which is a conjugate of estradiol and estrone-3-
sulfate which is a metabolite of estradiol and a conjugate of estrone
have been identified in WWTP influents in USA (Reddy et al., 2005).
According to Reddy et al. (2005), measured concentrations of steroid sul-
fates in the influent of a WWTP were 100 times greater than those of the
respective steroid glucuronides, suggesting that the preponderance of
glucuronides had dissociated prior reaching the treatment plant.
D'Ascenzo et al. (2003) attribute this dissociation to large amounts of
β-glucuronidase enzyme produced by fecal bacteria (Escherichia coli).
They also support that deconjugation continued in the sewer transit.
On the contrary, a small percentage of the steroid sulfates persisted
through biological treatment of sewage and was measured in the effluent.
6.9. Personal care products
Parabens, esters of 4-hydroxybenzoic acid, are extensively employed
as preservatives not only in a wide range of personal care products
919
(PCPs) but also in canned foods, beverages and pharmaceuticals
(Meyer et al., 2007; Lundov et al., 2009). The main concern once they
reach the environment is that they have proved to show estrogenic ac-
tivity (Routledge et al., 1998a,b; Golden et al., 2005; Soni et al., 2013),
relatively weak compared to that of 17b-estradiol but not negligible,
as they occur at much higher concentrations than the latter compound.
However, although the occurrence of parabens in wastewaters has
already been extensively studied (Lee et al., 2005; González-Mariño
et al., 2009; Gómez et al., 2010) limited data exists on the occurrence
of their TPs in wastewaters (Figs. S4 and S6). Moreover, it is noteworthy
that parabens can easily react with residual chlorine in tap water
(Canosa et al., 2006), yielding mostly mono and dichlorinated deriva-
tives and although halogenation masks the apparent estrogenic ac-
tivity of the parent compounds (Terasaki et al., 2009a), the resulting
chlorinated by-products show higher acute toxicity responses in the
D. magna test (Terasaki et al., 2009b). Some of the halogenated TPs
of methyl paraben have been detected in wastewaters. 3-Chloro-
methylparaben and 3,5-dichloro-methylparaben were detected in the
influents of WWTPs in Spain by González-Mariño et al. (2011), at con-
centrations ranging from 8 to 90 ng/L. These chlorinated derivatives
were also detected in some WWTP effluents but at lower concentrations
(up to 12 ng/L). The removals of the two chlorinated by-products after
the secondary treatment were slightly lower than those of their parent
compound suggesting a slightly higher persistence compared to their
precursor. However, both TPs exhibited a good removal efficiency
at 96.9% and 94.3% respectively. Moreover, González-Mariño et al.
(2011) also studied the biodegradability of these TPs and found that
they present slower biodegradation kinetics than their parent com-
pound, with half-lives of 3.3 days for the monochlorinated species and
8.6 days for the dichlorinated. This fact corroborates the findings on
the WWTP removal, where efficiency slightly decreases as chlorination
degree increases.
The methylation product of another PCP, triclosan, was also detected
in wastewaters (Lindstrom et al., 2002). Triclosan (5-chloro-2-(2,4-
dichlorophenoxy)phenol) is an important bactericide used in various
personal care (shampoo, toilet soap, deodorants, tooth paste) and con-
sumer (footwear, plastic wear) products. Methyl triclosan, a methyl
ether derivative, was detected in the influents and the effluents of
WWTPs in Switzerland. However, the higher concentrations (b2–
11 ng/L) that were observed at samples collected from the effluents
(compared to the parent compound) and the almost absence of this
TP from the influents (b1–4 ng/L) support the fact that methyl triclosan
is produced by triclosan probably via biological methylation despite the
fact that it is used as a starting material in the production of triclosan
(Lindstrom et al., 2002).
Moreover, triclosan may react during disinfection of waste-
water with free chlorine to form chlorinated triclosan derivatives
(CTDs) such as 4,5-dichloro-2-(2,4-dichlorophenoxy)phenol (4-Cl-
TCS), 5,6-dichloro-2-(2,4-dichlorophenoxy)phenol (6-Cl-TCS), and
4,5,6-trichloro-2-(2,4-dichlorophenoxy)phenol(4,6-Cl-TCS) (Canosa
et al., 2005; Rule et al., 2005). CTDs are of environmental concern be-
cause they may retain or possess enhanced levels of antimicrobial
and endocrine-disrupting properties, relative to triclosan. Additionally,
4-Cl-TCS, 6-Cl-TCS, and 4,6-Cl-TCS are known to undergo photolysis in
natural waters to form the respective dioxins, 2,3,7-trichlorodibenzo-p-
dioxin (2,3,7-TriCDD), 1,2,8-trichlorodibenzo-p-dioxin (1,2,8-TriCDD),
and 1,2,3,8-tetrachlorodibenzo-p-dioxin (1,2,3,8-TCDD) (Buth et al.,
2009). These products are very toxic and their toxicity/receptor binding
generally increases with chlorine substitution in the lateral positions
(Goldstein and Safe, 1989). Buth et al. (2011) have identified all three
chlorinated derivatives in WWTP influents and effluents in USA. Concen-
trations of 4-Cl-TCS, 6-Cl-TCS and 4,6-Cl-TCS in the influents were up to
17 ng/L, 98 ng/L and 29 ng/L, respectively. According to the authors the
observed concentrations in the influents of the municipal WWTP were
due to the reaction of triclosan with residual chlorine in the water mak-
ing up the influent stream in transport to the treatment facility. The CTDs
920 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
were removed at high rates, after the secondary treatment as expected
based on their increased hydrophobicity relative to triclosan, and
hence, higher tendency to sorb to sludge during the biological treatment
stage. The highest concentration of any CTD observed in the secondary
effluent samples was 3 ng/L. However, disinfection of the secondary ef-
fluent with sodium hypochloride led from one hand, to the increase of
the removal of triclosan but also on the other hand, to the formation of
CDTs in the final effluent due to the reaction of triclosan with free chlo-
rine during the disinfection step with concentrations up to 4, 9 and
22 ng/L for 4-Cl-TCS, 6-Cl-TCS and 4,6-Cl-TCS respectively. Moreover,
the authors demonstrated that in contrast to the free chlorine disinfec-
tion, the UV disinfection used by another WWTP did not generate any
CTDs while the influent CTDs were completely removed.
Finally, Mezcua et al. (2004) detected 2,7/2,8-dibenzodichloro-p-
dioxin, a photoproduct of triclosan, in WWTP influents and effluents
in Spain. The concentrations of dioxin in the influents were up to
8.9 μg/L while in the effluents ranged at lower values up to 0.4 μg/L,
thus indicating good removal after the secondary treatment. However,
total elimination was not observed.
6.10. Illicit drugs
The presence of IDs in environmental samples is becoming a matter
of global concern. The main source of these residues is mainly from con-
sumers, since after ingestion of a drug dose active parent compounds
and their metabolites are excreted through urine, entering urban waste-
water and reaching WWTPs. These substances may have low removal
efficiencies and persist in the treated wastewater (Chiaia et al., 2008;
Postigo et al., 2008; Zuccato and Castiglioni, 2009; Castiglioni et al.,
2011).
An interesting application of measuring IDs and their metabolites in
wastewater is “sewage epidemiology”, a novel approach for estimating
drug consumption in a community by wastewater analysis since it is
difficult to accurately estimate the use of IDs within communities
using current survey methods. The approach is based on the assumption
that all drug residues found in municipal wastewater derive from
human consumption. Consequently, the amount of drug or its metabo-
lite, determined in untreated municipal wastewater would reflect to its
collective consumption in a given local community. The correspondence
between the pattern of drug metabolites detected in wastewater and
their human excretion profile is another main assumption on which
the present method is based (Zuccato et al., 2008), but this correspon-
dence has only been assessed for a few compounds so far.
Cannabis (tetrahydrocannabinol, THC) is the most widely consumed
ID in the world. THC is almost completely metabolized in liver, produc-
ing (THC-COOH) and its glucuronide conjugate as the main urinary
metabolites. Thus, THC-COOH is widely used as a marker of consump-
tion and it has been detected in wastewaters (van Nuijs et al., 2011).
Its concentration in the influents (Table 4, see also Fig. S4) ranged up
to 489 ng/L (Bijlsma et al., 2012), while in the effluents (Table 4,
see also Fig. S6) was detected at much lower levels (b LOQ–72.8 ng/L),
exhibiting a good removal efficiency in the WWTPs. Postigo et al.
(2010) also identified OH-THC in the influents and effluents of
WWTPs in Spain. OH-THC is formed in the body after cannabis
consumption and is subsequently metabolized further to THC-COOH.
However, according to the authors (Postigo et al., 2010), cannabinoids
showed significant differences between the compounds, with elimina-
tion rates above 90% in the case of OH-THC, and poor removal (48%)
in the case of THC-COOH that presented often higher concentrations
at the WWTP outlet (after the secondary treatment). The poor removal
of THC-COOH that was observed by Postigo et al. (2010) might be
caused by deconjugation of less stable conjugated forms during the
treatment process.
Regarding cocainics, eight TPs of cocaine (COC) were detected in
wastewaters, including benzoylecgonine (BE), norbenzoylecgonine
(NBE), norcocaine (NCO),cocaethylene (CET), ecgonine methyl ester
(EME), ecgonine (ECG), anhydroecgonine (AEC), and anhydroecgonine
methyl ester (AME). According to metabolism studies, a small propor-
tion of COC (1–9% of the consumed dose) is not metabolized and is ex-
creted in the urine as unchanged drug. BE and EME are the primary
human metabolites of COC and they are excreted in urine at 45% and
40% (mean percentages) of the dose consumed, respectively (Baselt,
2004). In accordance with the known metabolism of COC in humans,
generally, higher concentrations of BE were measured compared with
the parent compound (Zuccato et al., 2005; Castiglioni et al., 2011;
Gheorghe et al., 2008; van Nuijs et al., 2009; Postigo et al., 2010). Also,
BE was the most abundant metabolite and was found in most samples
in the influents and the effluents of the WWTPs at concentrations rang-
ing up to 3701 ng/L and up to 4003 ng/L, respectively (see Table 4). The
removal of BE at the WWTPs after the secondary treatment was high
ranging from 88 to 98% (Huerta-Fontela et al., 2008; Postigo et al.,
2010). However, according to the findings of Bijlsma et al. (2009) al-
though on weekdays removal efficiency after the tertiary treatment
for BE was 95%, on weekends, when the highest concentrations were
reached, efficiency decreased to 35%. On the other hand, Gilart et al.
(2014) observed an increase of the BE concentration in the effluents
compared to the influents thus suggesting that conversion of conjugat-
ed metabolites to the original substances might have taken place after
the secondary treatment. Moreover, González-Mariño et al. (2012)
identified BE as the single product of hydrolysis of COC while NBE and
NCO were identified as chlorination products of COC. The authors dem-
onstrated that when chlorine is added, an oxidative N-dealkylation is
the main route, yielding NCOC, accompanied by a minor oxidative reac-
tion of the N-methyl group to produce N-formylnorcocaine (FNCOC).
However, hydrolysis also takes place, so that NBE is finally produced
by hydrolytic or chlorine mediated dealkylation of NCOC or BE,
respectively.
According to the urinary pattern of excretion the BE to COC ratio is
expected to be 5, since 45% is excreted as BE and only 9% is excreted
in the unchanged form assuming that there is no degradation of the
compound in the sewage. However, many researchers pointed out
that this ratio may exhibit fluctuations, from 0.8 to 3.2 (Metcalfe et al.,
2010; Bijlsma et al., 2012), suggesting the role of other sources of COC
probably related to transport, handling and consumption of the drug.
Castiglioni et al. (2011), also demonstrated the importance of the me-
tabolite AEC in the wastewaters since AEC is a metabolite of COC
when it is smoked and the AEC/BE ratio can be used for a rough estima-
tion of the amount of COC smoked in a community. However, the
metabolic pattern of COC when consumed as a crack depends on the py-
rolytic conditions and composition of the drug dose (Maurer et al.,
2006) and is therefore subjected to wide variability. Castiglioni et al.
(2011) found a small percentage of AEC (0.5% of the total metabolites)
which was expected, since only a small proportion of consumers may
use COC as crack; the majority is more likely to “snort” the drug.
Concerning morphine (MOR) residues, three metabolites/TPs were
detected, 6-acetyl morphine (6-AM), normorphine and morphine-3â-
D-glucuronide (M3G). The interpretation of the high MOR concentra-
tions in influents however remains difficult due to the presence of opi-
ate alkaloids in medicines and foods such as in analgesics and cough
suppression preparations or in poppy seeds, common ingredients in
bakery products (Cherny et al., 2010). These sources probably contrib-
ute to a large extent to the morphine burden and therefore mask hero-
ine consumption, since heroin is metabolically converted to morphine
inside the body. Nevertheless, the low levels of 6-AM (a metabolite of
heroin) clearly indicate that heroin consumption occurs. Concentration
levels of 6-AM in wastewater influents ranged up to 715 ng/L (Gilart
et al., 2014), while in most effluents its concentration was found to be
bLOQ (Table 4). High removal efficiency was estimated by Terzic et al.
(2010) for 6-AM in WWTPs after the secondary treatment that reaches
78%. Although morphine is excreted in urine mainly as glucuronide me-
tabolites, M3G was detected at low concentrations in WWTP influents
(2–18.1 ng/L) (Castiglioni et al., 2006), thus suggesting cleavage of the
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926 921

conjugated molecule in wastewater (Ternes, 1998; D'Ascenzo et al., A

00 000 500
Max
2003).

3 3
Mean
Moreover, 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP)
was also detected in wastewaters as a TP of methadone (METH). METH

25
is a synthetic opioid medically used as an analgesic and maintenance
anti-addictive and reductive preparation for use by patients with opioid
dependency (Fredheim et al., 2008). EDDP is the most important metab-

0
50
olite of METH and is formed by N-demethylation and cyclisation of

C(
METH in the liver. Concentrations of EDDP ranged between non-
detection to 206 ng/L and generally were higher than the concentrations

0
30
of the parent compound (Berset et al., 2010). EDDP was present in most
effluents samples, exhibiting very poor removal (3%) during treatment
(Terzic et al., 2010).

0
10
Codeine (COD), a natural opium alkaloid, is highly consumed either
by prescription or as an illicit drug (ID). COD is metabolized in the liver

TC
2O

C
T
O

-T

-T
and undergoes O-demethylation to form MOR and N-demethylation to

o-
H
C

or
Ep
Y-

dr
l
ch
ER

4-

hy
form norcodeine (NCOD). Wick et al. (2011) identified six metabolites/

i
Ep

an
i
4-
TPs of the COD in wastewaters. Codeinone, 14-hydroxycodeinone,

Ep
4-
NCOD, dihydrocodeine (DIC), neopine, and isoneopine are the TPs de-
tected. Wick et al. (2011) demonstrated that biological oxidation of co- B Utreated biosolids
deine leads to the formation of the R,β-unsaturated ketone codeinone, 16
Treated biosolids
which is the dominant TP of COD and is the precursor for further abiotic
15
and biotic transformation due to its high chemical reactivity leading to
the formation of the TPs identified (Martinez-Bueno et al., 2011a,b). 14
Codeinone was also identified in most influent samples, indicating
either transformation of COD to codeinone in the sewers and during 13
primary clarification or in human metabolism (Wick et al., 2011).
The higher concentrations of the TPs in the effluents compared to the 4
influents (Tables 2 and 4) underline the formation of the TPs after the
C(

secondary treatment. Hummel et al. (2006) found also high concen- 3

trations of DIC that reached 330 ng/L in the influents of WWTPs in
2
Germany.
Finally, lysergic acid diethylamide, abbreviated as LSD, also known 1
as lysergide (INN) and colloquially as acid, is a semisynthetic psychedel-

BZ
BZ

BZ
ic drug of the ergoline family. It is used mainly as an entheogen, recrea-

-C
-C

-C

H
H

H
tional drug, and as an agent in psychedelic therapy. However, only one

iO
2O

3O

report exists concerning the occurrence of the TPs of LSD in wastewater.
Postigo et al. (2010) identified nor-LSD, a N-demethylated product as a
TP of LSD in wastewater influents and effluents of WWTPs in Spain. Nor-
LSD exhibiting only 7% frequency of detection in influents and 14% in
wastewater effluents at concentration ranged at low levels, 4.3 ng/L
and 0.4 ng/L respectively, while the parent compound was not detected.
7. Occurrence of TPs of PPCPs and IDs in biosolids and sludge
Although there is a vast body of knowledge about the concentrations
of PPCPs in the aquatic environment, only a few reports have been pub-
lished on environmental concentrations in both sewage and sludge.
Even more, too little is known about the occurrence and fate of their
TPs. There is still a lack of fundamental data and its detection in solid
samples still remains a limiting part in environmental analysis, mainly
due to the lack of suitable selective and sensitive analytical methods.
Based to the authors knowledge, only a handful specialized reports are
published (Miao et al., 2005; Xu et al., 2007; McClellan and Halden,
2010). The obtained results suggest (Fig. 3) that sewage sludge and bio-
solids can serve as the main reservoir of TP residues underling the im-
portance of sludge management strategies.
8. Concluding remarks and future outlook
In this review, the occurrence and fate of the TPs of PPCPs and IDs in
WWTPs is reported. The aim of the study was to provide an integrated
knowledge on the identity of the TPs that have been detected in the
influents and the effluents of WWTPs, their concentration levels and
finally their removal.
D
Fig. 3. Concentrations of TPs of pharmaceuticals: (Α) in sludge (μg/g); (B) in biosolids
(μg/kg dry weight) (Miao et al., 2005; Xu et al., 2007; McClellan and Halden, 2010).
Generally, more scientific data exists on antibiotics, NSAIDs, analge-
sics, antiepileptics, antidepressants and IDs. The most studied parent
compound is carbamazepine for which nine TPs have been detected in
WWTP influents and effluents. On the contrary, scarce data exists on
TPs of antiviral, anticancer, antidiabetic, psychoactive, antilipidemic
drugs and other groups of drugs.
Most investigations are conducted in Europe and results are mainly
available from Germany and Spain. North-American countries (USA and
Canada) also contributed to this awareness, whereas scarce data for
other regions around the world are limited to China and Australia.
Concentrations of the TPs in the wastewaters usually range in
the ng/L level, however exceptions like ERY-H2O, SA, CLF acid, OH-
IBU, CX-IBU, 4-AAA, 4-MAA, 4-AA, 4-FAA, O-DES-Me-VNF, OH-BBP,
erythro/threo-hydrobupropion, CBZ-Ep, DiOH-CBZ, cotinine, and BE
usually reach the μg/L level.
Most TPs detected are metabolites formed after human consump-
tion. However, TPs may stem from other abiotic processes like hydro-
lysis (for example, ERY-H2O) or photolysis (like dioxin, the TP of
triclosan). Nevertheless, biotic transformations may also take place in
the WWTP leading to the occurrence of TPs in the effluents (like
the TPs of codeine). The tertiary treatment with chlorine disinfection
may also cause structural changes of the remaining PPCPs and their
TPs that survived biological treatment, leading to the formation of
chlorinated-TPs (like the chlorination products of methyl paraben and
triclosan) in the final effluent and consequently to the receiving waters.
922 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
In most cases concentrations of TPs in the effluents were smaller
than those in the influents thus exhibiting removals in WWTPs that
ranged from 0 to 100%. However, some compounds were present at
higher concentrations in the outflow of WWTP indicating their forma-
tion from the parent compound or their release from conjugates during
the treatment process.
No specific pattern is observed in the occurrence of TPs of PPCPs and
IDs in municipal, industrial and hospital wastewaters. On the contrary,
great discrepancies are observed between countries indicating the
different consumption rates. Variations are also observed within
the same country, between different WWTPs. This can be attributed to
the different pharmaceutical prescription patterns in a country which
determine or affect influent concentrations and to the removal efficien-
cies and the treatment applied at individual WWTPs, which affect efflu-
ent concentrations. In general, due to the dynamic correlation of the
parent compounds with their TPs, the investigation of parent com-
pound/metabolite concentration patterns will be a valuable tool in the
further assessment of the environmental fate of PPCPs and IDs.
Based on the remarks outlined above, it is clear that more informa-
tion is now available on occurrence and fate of TPs of PPCPs and IDs in
WWTPs. However, knowledge in this field is in its formative stage and
there are still many knowledge gaps on various aspects of TPs and me-
tabolites in the fields of occurrence, fate, exposure, toxicity and risks. For
example, most of the studied works are focused on aqueous matrices by
screening “known” TPs and metabolites. Therefore, from a monitoring
performance point of view there is a need to enlarge the number of
TPs that should be screened in the typical target analysis. Furthermore,
measurements in biosolids and sludge for their occurrence and fate in
WWTPs are needed. Detailed case studies of untreated and treated
sludge in pilot WWTPs or in actual wastewater matrices and full-scale
WWTPs are required. Incorporating the occurrence information of TPs
on aqueous and solid phase will improve knowledge regarding their
fate in WWTPs. Finally, further investigations on the ecotoxicological
potential of target TPs and their mixtures in wastewaters effluents are
indispensable. A battery of biological testing tools would be applied as
a first tier to address the integrated toxicity of these complex samples.
In continuation of this aspect, approaches to prioritize TPs of PPCPs
with regard to toxicological significance should be also developed inte-
grating the existing research in this field.
Acknowledgments
The support for this study was received from the Greek Ministry
of Education and Religious Affairs through Operational Program
“Education and Lifelong Learning” of the National Strategic Reference
Framework (NSRF)-Research Funding Program “Excellence II (Aristeia
II)” under the title “Advanced microextraction approaches based on
novel nano-polymers to measure pharmaceuticals, personal care prod-
ucts and their transformation products in the aquatic environment”
(No. 4199), which is gratefully appreciated.
Appendix A. Supplementary data
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.scitotenv.2014.10.021.
References
Bahlmann A, Brack W, Schneider RJ, Krauss M. Carbamazepine and its metabolites in
wastewater: analytical pitfalls and occurrence in Germany and Portugal. Water Res
2014;57:104–14.
Barclay VKH, Tyrefors NL, Johansson IM, Pettersson CE. Chiral analysis of metoprolol and
two of its metabolites, a-hydroxymetoprolol and deaminated metoprolol, in waste-
water using liquid chromatography–tandem mass spectrometry. J Chromatogr
2012;1269:208–17.
Baselt RC. Disposition of toxic drugs and chemicals in man. Foster City, California, USA:
Biomedical Publications; 2004.
Batt AL, Mitch SK, Lazorchak MJ. Analysis of ecologically relevant pharmaceuticals in
wastewater and surface water using selective solid-phase extraction and UPLC–MS/
MS. Anal Chem 2008;80:5021–30.
Baumann P. Care of depression in the elderly: comparative pharmacokinetics of SSRIs. Int
Clin Psychopharmacol 1998;13:S35–43.
Behera SK, Kim HW, Oh J-E, Park H-S. Occurrence and removal of antibiotics, hormones
and several other pharmaceuticals in wastewater treatment plants of the largest in-
dustrial city of Korea. Sci Total Environ 2011;409:4351–60.
Bendz D, Paxéus NA, Ginn TR, Loge FJ. Occurrence and fate of pharmaceutically active
compounds in the environment, a case study: Höje River in Sweden. J Hazard
Mater 2005;122:195–204.
Benfield P, Heel RC, Lewis SP. Fluoxetine: a review of its pharmacodynamic and pharma-
cokinetic properties, and therapeutic efficacy in depressive illness. Drugs 1986;32:
481–508.
Benotti MJ, Brownawell BJ. Distributions of pharmaceuticals in an urban estuary during
both dry- and wet-weather conditions. Environ Sci Technol 2007;41:5795–802.
Berset JD, Brenneisen R, Mathieu C. Analysis of licit and illicit drugs in waste, surface and
lake water samples using large volume direct injection high performance liquid chro-
matography–electrospray tandem mass spectrometry HPLC–MS/MS. Chemosphere
2010;81:859–66.
Bijlsma L, Sancho JV, Pitarch E, Ibáñez M, Hernádez F. Simultaneous ultra high pressure
liquid chromatography–mass spectrometry determination of amphetamine like
stimulants, cocaine and its metabolites, and a cannabis metabolite in surface water
and urban wastewater. J Chromatogr A 2009;1216:3078–89.
Bijlsma L, Emke E, Hernández F, De Voogt P. Investigation of drugs of abuse and relevant
metabolites in Dutch sewage water by liquid chromatography coupled to high reso-
lution mass spectrometry. Chemosphere 2012;89:1399–406.
Bleeker EAJ, Van der Geest HG, Klamer HJC, De Voogt P, Wind E, Kraak MHS. Toxic and
genotoxic effects of azaarenes: isomers and metabolites. Polycyclic Aromat Compd
1999;13:191–203.
Boczar BA, Forney LJ, Begley WM, Larson RJ, Federle TW. Characterization and distribution
of esterase activity in activated sludge. Water Res 2001;35:4208–16.
Boix C, Ibáñez M, Zamora T, Sancho JV, Niessen WMA, Hernández F. Identification of new
omeprazole metabolites in wastewaters and surface waters. Sci Total Environ 2014;
468–469:706–14.
Boleda MR, Galceran MT, Ventura F. Trace determination of cannabinoids and opiates in
wastewater and surface waters by ultra-performance liquid chromatography–
tandem mass spectrometry. J Chromatogr A 2007;1175:38–48.
Bones J, Kevin VT, Paull B. Using environmental analytical data to estimate levels of com-
munity consumption of illicit drugs and abused pharmaceuticals. J Environ Monit
2007;9:701–7.
Bort R, Ponsoda X, Carrasco E, Gomez-Lechon MJ, Castell JV. Comparative metabolism of
the nonsteroidal antiinflammatory drug, aceclofenac, in the rat, monkey, and
human. Drug Metab Dispos 1996a;24:969–75.
Bort R, Ponsoda X, Carrasco E, Gómez-Lechón MJ, Castell JV. Metabolism of aceclofenac in
humans. Drug Metab Dispos 1996b;24:834–41.
Boxall ABA, Sinclair CJ, Fenner K, Koplin D, Maund SJ. Peer reviewed: when synthetic
chemicals degrade in the environment. Environ Sci Technol 2004;38:368–75.
Bradley PM, Barber LB, Kolpin DW, McMahon PB, Chapelle FH. Biotransformation of caf-
feine, cotinine, and nicotine in stream sediments: implications for use as wastewater
indicators. Environ Toxicol Chem 2007;26:1116–21.
Bramer SL, Kallungal BA. Clinical considerations in study designs that use cotinine as a
biomarker. Biomarkers 2003;8:187–203.
Breton H, Cociglio M, Bressolle F, Peyriere H, Blayac JP, Hillaire-Buys D. Liquid
chromatography–electrospray mass spectrometry determination of carbamazepine,
oxcarbazepine and eight of their metabolites in human plasma. J Chromatogr B
2005;828:80–90.
Bueno Martínez MJ, Agüera A, Gómez MJ, Hernando MD, García-Reyes JF, Fernández-Alba
AR. Application of liquid chromatography/quadrupole-linear ion trap mass spec-
trometry and time-of-flight mass spectrometry to the determination of pharmaceuti-
cals and related contaminants in wastewater. Anal Chem 2007;79:9372–84.
Buerge IJ, Kahle M, Buser HR, Muller MD, Poiger T. Nicotine derivatives in wastewater and
surface waters: application as chemical markers for domestic wastewater. Environ
Sci Technol 2008;42:6354–60.
Buser H-R, Poiger T, Muller MD. Occurrence and environmental behaviour of the chiral
pharmaceutical drug ibuprofen in surface waters and in wastewater. Environ Sci
Technol 1999;33:2529–35.
Buth JM, Grandbois M, Vikesland PJ, McNeill K, Arnold WA. Aquatic photochemistry of
chlorinated triclosan derivatives: potential source of polychlorodibenzo-p-dioxins.
Environ Toxicol Chem 2009;28:2555–63.
Buth JM, Ross MR, McNeill K, Arnold WA. Reprint of: removal and formation of chlorinat-
ed triclosan derivativesin wastewater treatment plants using chlorine and UV disin-
fection. Chemosphere 2011;85:284–9.
Calza P, Debora F. Advanced mass spectrometry-based techniques for the identification
and structure elucidation of transformation products of emerging contaminants. In.
In: Lambropoulou DA, Nollet LML, editors. Transformation products of emerging con-
taminants in the environment: analysis, processes, occurrence, effects and risks. 1st
ed. John Wiley and Sons Ltd; 2014. p. 325–46.
Canosa P, Morales S, Rodriguez I, Rubi E, Cela R, Gomez M. Aquatic degradation of triclo-
san and formation of toxic chlorophenols in presence of low concentrations of free
chlorine. Anal Bioanal Chem 2005;383:1119–26.
Canosa P, Rodrıguez I, Rubı E, Negreira N, Cela R. Formation of halogenated by-products of
parabens in chlorinated water. Anal Chim Acta 2006;575:106–13.
Carlsson C, Johansson A, Alvan B, Bergman K, Kuhler T. Are pharmaceuticals potent envi-
ronmental pollutants? Part I: environmental risk assessments of selected active phar-
maceutical ingredients. Sci Total Environ 2006;364:67e87.
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
Carretero I, Vadillo M, Laserna JJ. Determination of antipyrine metabolites in human plas-
ma by solid-phase extraction and micellar liquid chromatography. Analyst 1995;120:
1729–32.
Castiglioni S, Zuccato E, Crisci E, Chiabrando C, Fanelli R, Bagnati R. Identification and mea-
surement of illicit drugs and their metabolites in urban wastewater by liquid chroma-
tography–tandem mass spectrometry. Anal Chem 2006;78:8421–9.
Castiglioni S, Bagnati R, Melis M, Panawennage D, Chiarelli P, Fanelli R, et al. Identification
of cocaine and its metabolites in urban wastewater and comparison with the human
excretion profile in urine. Water Res 2011;45:5141–50.
Celiz MD, Tso J, Aga DS. Pharmaceutical metabolites in the environment: analytical chal-
lenges and ecological risks. Environ Toxicol Chem 2009;28:2473–84.
Chee-Sanford JC, Aminov RI, Krapac IJ, Garriques-Jeanjean N, Mackie RI. Occurrence and
diversity of tetracycline resistance genes in lagoons and groundwater underlying
two swine production facilities. Appl Environ Microbiol 2001;67:1494–502.
Cherny NI, Baselga J, de Conno F, Radbruch L. Formulary availability and regulatory bar-
riers to accessibility of opioids for cancer pain in Europe: a report from the ESMO/
EAPC Opioid Policy. Init Ann Oncol 2010;21:615–26.
Chiaia AC, Banta-Green C, Field J. Eliminating solid phase extraction with large-volume in-
jection LC/MS/MS: analysis of illicit and legal drugs and human urine indicators in
U.S. wastewaters. Environ Sci Technol 2008;42:8841–8.
Chiaia-Hernandez AC, Schymanski EL, Kumar P, Singer HP, Hollender J. Suspect and non-
target screening approaches to identify organic contaminant records in lake sedi-
ments. Anal Bioanal Chem 2014. http://dx.doi.org/10.1007/s00216-014-8166-0. [in
Press].
Cruz-Morato C, Ferrando-Climent L, Rodriguez-Mozaz S, Barcelo D, Marco-Urrea E, Vicent
Sarra M. Degradation of pharmaceuticals in non-sterile urban wastewater by
Trametes versicolor in a fluidized bed bioreactor. Water Res 2013;47:5200–10.
Custodio JBA, Dinis TCP, Almeida LM, Madeira VMC. Tamoxifen and hydroxytamoxifen as
intramembraneous inhibitors of lipid peroxidation. Evidence for peroxyl radical scav-
enging activity. Biochem Pharmacol 1994;47:1989–98.
D'Ascenzo G, Di Corcia A, Gentili A, Mancini R, Mastropasqua R, Nazzari M, et al. Fate of
natural estrogen conjugates in municipal sewage transport and treatment facilities.
Sci Total Environ 2003;302:199–209.
Dong Z, Senn DB, Moran RE, Shine JP. Prioritizing environmental risk of prescription phar-
maceuticals. Regul Toxicol Pharmacol 2013;65:60–7.
Escher BI, Fenner K. Recent advances in environmental risk assessment of transformation
products. Environ Sci Technol 2011;45:3835–47.
Fatta-Kassinos D, Vasquez MI, Kümmerer K. Transformation products of pharmaceuticals
in surface waters and wastewater formed during photolysis and advanced oxidation
processes — degradation, elucidation of byproducts and assessment of their biological
potency. Chemosphere 2011;85:693–709.
Feldmann DF, Zuehlke S, Heberer T. Occurrence, fate and assessment of polar metamizole
dipyrone; residues in hospital and municipal wastewater. Chemosphere 2008;71:
1754–64.
Ferrando-Climent L, Collado N, Buttiglieri G, Gros M, Rodriguez-Roda I, Rodriguez-Mozaz
S, et al. Comprehensive study of ibuprofen and its metabolites in activated sludge
batch experiments and aquatic environment. Sci Total Environ 2012;438:404–13.
Ferrando-Climent L, Rodriguez-Mozaz S, Barceló D. Development of a UPLC–MS/MS
method for the determination of ten anticancer drugs in hospital and urban waste-
waters, and its application for the screening of human metabolites assisted by
information-dependent acquisition tool IDA; in sewage samples. Anal Bioanal Chem
2013;405:5937–52.
Ferrer I, Thurman EM. Identification of a new antidepressant and its glucuronide metab-
olite in water samples using liquid chromatography/quadrupole time-of-flight mass
spectrometry. Anal Chem 2010;82:8161–8.
Fredheim OM, Moksnes K, Borchgrevink PC, Kaasa S, Dale O. Clinical pharmacology of
methadone for pain. Acta Anaesthesiol Scand 2008;52:879–89.
Gentili A, Perret D, Marchese S. Analysis of free estrogens and their conjugates in sewage
and river waters by solid phase extraction then liquid chromatography–
electrospray–tandem mass spectrometry. Chromatographia 2002;56:25–32.
Gheorghe A, van Nuijs A, Pecceu B, Bervoets L, Jorens PG, Blust R, et al. Analysis of cocaine
and its principal metabolites in waste and surface water using solid-phase extraction
and liquid chromatography–ion trap tandem mass spectrometry. Anal Bioanal Chem
2008;391:1309–19.
Ghoshdastidar AJ, Fox S, Tong AZ. The presence of the top prescribed pharmaceuticals in
treated sewage effluents and receiving waters in Southwest Nova Scotia, Canada. En-
viron Sci Pollut Res 2014. http://dx.doi.org/10.1007/s11356-014-3400-z.
Gilart N, Cormack PAG, Marcé RM, Fontanals N, Borrull F. Selective determination of phar-
maceuticals and illicit drugs in wastewaters using a novel strong cation-exchange
solid-phase extraction combined with liquid chromatography–tandem mass spec-
trometry. J Chromatogr A 2014;1325:137–46.
Glassmeyer ST, Furlong ET, Koplin DW, Cahill JD, Zaugg SD, Werner SL, et al. Transport of
chemical and microbial compounds from known wastewater discharges: potential
use as indicators of human fecal contamination. Environ Sci Technol 2005;39:5157–69.
Golden R, Gandy J, Vollmer G. A review of the endocrine activity of parabens and implica-
tions for potential risks to human health. Crit Rev Toxicol 2005;35:435–58.
Goldstein JA, Safe SH. Mechanisms of action and structure–activity relationship for the
chlorinated dibenzo-p-dioxins and related compounds. In: Kimbrough RD, Jensen
AA, editors. Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and
related products. Amsterdam: Elsevier; 1989. p. 239–93.
Gómez MJ, Martínez Bueno MJ, Lacorte S, Fernández-Alba AR, Agüera A. Pilot survey mon-
itoring pharmaceuticals and related compounds in a sewage treatment plant located
on the Mediterranean coast. Chemosphere 2007;66:993–1002.
Gómez MJ, Sirtori C, Mezcua M, Fernández-Alba AR, Agüera A. Photodegradation study of
three dipyrone metabolites in various water systems: identification and toxicity of
their photodegradation products. Water Res 2008;42:2698–706.
923
Gómez MJ, Gómez-Ramos MM, Malato O, Mezcua M, Fernández-Alba AR. Rapid automat-
ed screening, identification and quantification of organic microcontaminants and
their main transformation products in wastewater and river waters using liquid chro-
matography quadrupole-time-of-flight mass spectrometry with an accurate-mass
database. J Chromatogr A 2010;1217:7038–54.
Gómez-Ramos M, Pérez-Parada A, García-Reyes JF, Fernández-Alba AR, Agüera A. Use of
an accurate-mass database for the systematic identification of transformation prod-
ucts of organic contaminants in wastewater effluents. J Chromatogr A 2011;1218:
8002–12.
González-Mariño I, Quintana JB, Rodríguez I, Cela R. Simultaneous determination of
parabens, triclosan and triclocarban in water by liquid chromatography/electrospray
ionisation tandem mass spectrometry. Rapid Commun Mass Spectrom 2009;23:
1756–66.
González-Mariño I, Quintana JB, Rodríguez I, Cela R. Evaluation of the occurrence and
biodegradation of parabens and halogenated by-products in wastewater by
accurate-mass liquid chromatography–quadrupole-time-of-flight-mass spectrome-
try LC–QTOF-MS. Water Res 2011;45:6770–80.
González-Mariño I, Quintana JB, Rodríguez I, Sánchez-Méndez N, Cela R. Transformation
of cocaine during water chlorination. Anal Bioanal Chem 2012;404:3135–44.
Gros M, Rodríguez-Mozaz S, Barceló D. Fast and comprehensive multi-residue analysis of
a broad range of human and veterinary pharmaceuticals and some of their metabo-
lites in surface and treated waters by ultra-high-performance liquid chromatography
coupled to quadrupole-linear ion trap tandem mass spectrometry. J Chromatogr A
2012;1248:104–21.
Gulkowska A, Leung HW, So MK, Taniyasu S, Yamashita N, Yeung LWY, et al. Removal of
antibiotics from wastewater by sewage treatment facilities in Hong Kong and
Shenzhen, China. Water Res 2008;42:395–403.
Halling-Sørensen B, Nielsen N, Lansky PF, Ingerslev F, Hansen L, Lützhøft HC, et al. Occur-
rence, fate and effects of pharmaceutical substances in the environment — a review.
Chemosphere 1998;36:357–94.
Haufroid V, Lison D. Urinary cotinine as a tobacco smoke exposure index: a mini review.
Int Arch Occup Environ Health 1998;71:162–3.
Heberer T. Occurrence, fate, and removal of pharmaceutical residues in aquatic environ-
ment: a review of recent research data. Toxicol Lett 2002;131:5–17.
Heberer T, Stan H-J. Determination of clofibric acid and N-phenylsulfonyl;-sarcosine in
sewage, river and drinking water. Int J Environ Anal Chem 1997;67:113–24.
Henschel K, Wenzel A, Diedrich M, Fliedner A. Environmental hazard assessment of phar-
maceuticals. Regul Toxicol Pharmacol 1997;25:220–5.
Hirsch R, Ternes T, Haberer K, Kratz K-L. Occurrence of antibiotics in the aquatic environ-
ment. Sci Total Environ 1999;225:109–18.
Holčapek M, Kolářová L, Nobilis M. High-performance liquid chromatography–tandem
mass spectrometry in the identification and determination of phase I and phase II
drug metabolites. Anal Bioanal Chem 2008;391:59–78.
Huerta-Fontela M, Galceran MT, Ventura F. Ultra performance liquid chromatography–
tandem mass spectrometry analysis of stimulatory drugs of abuse in wastewater
and surface waters. Anal Chem 2007;79:3821–9.
Huerta-Fontela M, Galceran MT, Martin-Alons J, Ventura F. Occurrence of psychoactive
stimulatory drugs in wastewater in north-eastern Spain. Sci Total Environ 2008;
397:21–40.
Huerta-Fontela M, Galceran MT, Ventura F. Fast liquid chromatography–quadrupole-
linear ion trap mass spectrometry for the analysis of pharmaceuticals and hormones
in water resources. J Chromatogr A 2010;1217:4212–22.
Hummel D, Löffler D, Fink G, Ternes TA. Simultaneous determination of psychoactive
drugs and their metabolites in aqueous matrices by liquid chromatography mass
spectrometry. Environ Sci Technol 2006;40:7321–8.
Jiang JQ, Yin Q, Zhou JL, Pearce P. Occurrence and treatment trials of endocrine disrupting
chemicals EDCs; in wastewaters. Chemosphere 2005;61:544–50.
Jones OAH, Voulvoulis N, Lester JN. Human pharmaceuticals in wastewater treatment
processes. Crit Rev Environ Sci Technol 2005;35:401–27.
Kaiser E, Prasse C, Broder K, Ternes T. Transformation of three human metabolites of car-
bamazepine during sand filtration. SETAC Europe 23rd Annual Meeting, Glasgow;
2013.
Karthikeyan KG, Meyer MT. Occurrence of antibiotics in wastewater treatment facilities in
Wisconsin, USA. Sci Total Environ 2006;361:196–207.
Kaye CM, Haddock RE, Langley PF, Mellows G, Tasker TCG, Greb WH. A review of the me-
tabolism and pharmacokinetics of paroxetine in man. Acta Psychiatr Scand 1989;80:
60–75.
Kazner C, Lehnberg K, Kovalova L, Wintgens T, Melin T, Hollender J, et al. Removal of endo-
crine disruptors and cytostatics from effluent by nanofiltration in combination with
adsorption on powdered activated carbon. Water Sci Technol 2008;58:1699–706.
Kepp DR, Sidelmann UG, Tjornelund J, Hansen SH. Simultaneous quantitative determina-
tion of the major phase I and II metabolites of ibuprofen in biological fluids by high-
performance liquid chromatography on dynamically modified silica. J Chromatogr B
Biomed Sci Appl 1997;696:235–41.
Kern S, Baumgartner R, Helbling DE, Hollender J, Singer H, Loos MJ, et al. A tiered proce-
dure for assessing the formation of biotransformation products of pharmaceuticals
and biocides during activated sludge treatment. J Environ Monit 2012;12:2100–11.
Khetan SK, Collins TJ. Human pharmaceuticals in the aquatic environment: a challenge to
green chemistry. Chem Rev 2007;107:2319–64.
Kormos JL, Schulz M, Wagner M, Ternes TA. Multistep approach for the structural identi-
fication of biotransformation products of iodinated X-ray contrast media by liquid
chromatography/hybrid triple quadrupole linear ion trap mass spectrometry and
1
H and 13C nuclear magnetic resonance. Anal Chem 2009;81:9216–24.
Kormos JL, Schulz M, Kohler H-PE, Ternes TA. Document biotransformation of selected
iodinated X-ray contrast media and characterization of microbial transformation
pathways. Environ Sci Technol 2010;44:4998–5007.
924 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
Kormos JL, Schulz M, Ternes TA. Occurrence of iodinated X-ray contrast media and their
biotransformation products in the urban water cycle. Environ Sci Technol 2011;45:
8723–32.
Kosma CI, Lambropoulou DA, Albanis TA. Occurrence and removal of PPCPs in municipal
and hospital wastewaters in Greece. J Hazard Mater 2010;179:804–17.
Kosma CI, Lambropoulou DA, Albanis TA. Investigation of PPCPs in wastewater treatment
plants in Greece: occurrence, removal and environmental risk assessment. Sci Total
Environ 2014a;466–467:421–38.
Kosma CI, Lambropoulou DA, Albanis TA. Comprehensive study of the antidiabetic drug
metformin and its transformation product guanylurea in Greek wastewaters; 2014b
[submitted for publication].
Krauss M, Singer H, Hollender J. LC-high resolution MS in environmental analysis: from
target screening to the identification of unknowns. Anal Bioanal Chem 2010;397:
943–51.
Lajeunesse A, Gagnon C, Sauvé S. Determination of basic antidepressants and their N-
desmethyl metabolites in raw sewage and wastewater using solid phase extraction
and liquid chromatography–tandem mass. Anal Chem 2008;80:5325–33.
Lambropoulou DA, Nollet LML. Transformation products of emerging contaminants in the
environment: analysis, processes, occurrence, effects and risks. 1st ed. John Wiley and
Sons Ltd; 2014.
Lapworth DJ, Baran N, Stuart ME, Ward RS. Emerging organic contaminants in groundwa-
ter: a review of sources, fate and occurrence. Environ Pollut 2012;163:287–303.
Leclercq M, Mathieu O, Gomez E, Casellas C, Fenet H, Hillaire-Buys D. Presence and fate of
carbamazepine, oxcarbazepine, and seven of their metabolites at wastewater treat-
ment plants. Arch Environ Contam Toxicol 2009;56:408–15.
Lee HB, Sarafin K, Peart TE, Svoboda ML. Acidic pharmaceuticals in sewage — methodol-
ogy, stability test, occurrence, and removal from Ontario samples. Water Qual Res J
Can 2003;38:667–82.
Lee HB, Peart TE, Svoboda ML. Determination of endocrine-disrupting phenols, acidic
pharmaceuticals, and personal-care products in sewage by solid-phase extraction
and gas chromatography–mass spectrometry. J Chromatogr A 2005;1094:122–9.
Lenz K, Mahnik S, Weissenbacher N, Mader RM, Krenn P, Hann S, et al. Monitoring, re-
moval and risk assessment of cytostatic drugs in hospital wastewater. Water Sci
Technol 2007;56:141–9.
Lertratanangkoon K, Horning MG. Metabolism of carbamazepine. Drug Metab Dispos
1982;10:1–10.
Letzel M, Weiss K, Schüssler W, Sengl M. Occurrence and fate of the human pharmaceu-
tical metabolite ritalinic acid in the aquatic system. Chemosphere 2010;81:1416–22.
Leung HW, Jin L, Wei S, Tsui MMP, Zhou B, Jiao L, et al. Pharmaceuticals in tap water:
human health risk assessment and proposed monitoring framework in China.
Environ Health Perspect 2013;121:839–46.
Li Y, Zhu G, Nq WJ, Tan SK. A review on removing pharmaceutical contaminants from
wastewater by constructed wetlands: design, performance and mechanism. Sci
Total Environ 2014;468–469:908–32.
Li Z, Fenet H, Gomez E, Chiron S. Transformation of theantiepileptic drug oxcarbazepine
upon different waterdisinfection processes. Water Res 2011;45:1587–96.
Lin AYC, Tsai YT. Occurrence of pharmaceuticals in Taiwan's surface waters: impact of
waste streams from hospitals and pharmaceutical production facilities. Sci Total
Environ 2009;407:3793–802.
Lin AYC, Lee WN, Wang XH. Ketamine and the metabolite norketamine: persistence and
phototransformation toxicity in hospital wastewater and surface water. Water Res
2014;53:351–60.
Lindberg RH, Wennberg P, Johansson MI. Screening of human antibiotic substances and
determination of weekly mass flows in five sewage treatment plants in Sweden.
Environ Sci Technol 2005;39:3421–9.
Lindstrom M, Buerge IJ, Poiger T, Bergqvist P-A, Muller M, Buser H-R. Occurrence and en-
vironmental behavior of the bactericide triclosan and its methyl derivative in surface
waters and in wastewater. Environ Sci Technol 2002;36:2322–9.
Lundov MD, Moesby L, Zachariae C, Johansen JD. Contamination versus preservation of
cosmetics: a review on legislation, usage, infections, and contact allergy. Contact
Dermatitis 2009;60:70–8.
Luo Y, Guo W, Ngo HH, Nghiem LD, Hai FI, Zhang J, et al. A review on the occurrence of
micropollutants in the aquatic environment and their fate and removal during waste-
water treatment. Sci Total Environ 2014;473–474:619–41.
Lynn RK, Smith RG, Thompson RM, Deinzer ML, Griffin D, Gerber N. Characterization of
glucuronide metabolites of carbamazepine in human urine by gas chromatography
and mass spectrometry. Drug Metab Dispos 1978;6:494–501.
Maggs JL, Pirmohamed M, Kitteringham NR, Park BK. Characterization of the metabolites
of carbamazepine in patient urine by liquid chromatography/mass spectrometry.
Drug Metab Dispos 1997;25:275–80.
Marciniec B. Photochemical decomposition of phenazone derivatives. Part 3: kinetics of
photolysis in aqueous solutions. Pharmazie 1984;39:103–6.
Martinez-Bueno MJM, Uclés S, Hernando MD, Fernández-Alba AR. Development of a sol-
vent free method for the simultaneous identification/quantification of drugs of abuse
and their metabolites in environmental water by LC–MS/MS. Talanta 2011a;85:
157–66.
Martinez-Bueno MJM, Uclés S, Hernando MD, Dávoli E, Fernández-Alba AR. Evaluation of
selected ubiquitous contaminants in the aquatic environment and their transforma-
tion products. A pilot study of their removal from a sewage treatment plant. Water
Res 2011b;45:2331–41.
Maurer HH, Sauer C, Theobald DS. Toxicokinetics of drugs of abuse: current knowledge of
the isoenzymes involved in the human metabolism of tetrahydrocannabinol, cocaine,
heroin, morphine, and codeine. Ther Drug Monit 2006;28:447–53.
McArdell CS, Molnar E, Suter MJF, Giger W. Occurrence and fate of macrolide antibiotics in
wastewater treatment plants and in the Glatt Valley Watershed, Switzerland. Environ
Sci Technol 2003;24:5479–86.
McClellan K, Halden RU. Pharmaceuticals and personal care products in archived U.S. bio-
solids from the 2001 EPA national sewage sludge survey. Water Res 2010;44:658–68.
Méndez-Arriaga F, Esplugas S, Giménez J. Photocatalytic degradation of non-steroidal
anti-inflammatory drugs with TiO2 and simulated solar irradiation. Water Res
2008;42:585–94.
Metcalfe CD, Miao X-S, Koenig BG, Struger J. Distribution of acidic and neutral drugs in
surface water near sewage treatment plants in the lower Great Lakes, Canada. Envi-
ron Toxicol Chem 2003;22:2881–9.
Metcalfe CD, Chu S, Judt C, Li H, Oakes KD, Servos MR, et al. Antidepressants and their me-
tabolites in municipal wastewater, and downstream exposure in an urban watershed.
Environ Toxicol Chem 2010;29:79–89.
Meyer BK, Ni A, Hu B, Shi L. Antimicrobial preservative use in parenteral products: past
and present. J Pharm Sci 2007;96:3155–67.
Mezcua M, Gómez MJ, Ferrer I, Aguera A, Hernando MD, Fernández-Alba AR. Evidence of
2,7/2,8-dibenzodichloro-p-dioxin as a photodegradation product of triclosan in water
and wastewater samples. Anal Chim Acta 2004;524:241–7.
Miao XS, Metcalfe CD. Determination of carbamazepine and its metabolites in aqueous
samples using liquid chromatography–electrospray tandem mass spectrometry.
Anal Chem 2003;75:3731–8.
Miao XS, Koenig BG, Metcalfe CD. Analysis of acidic drugs in the effluents of sewage treat-
ment plants using liquid chromatography–electrospray ionization tandem mass
spectrometry. J Chromatogr A 2002;952:139–47.
Miao XS, Bishay F, Chen M, Metcalfe CD. Occurrence of antimicrobials in the final effluents
of wastewater treatment plants in Canada. Environ Sci Technol 2004;38:3533–41.
Miao XS, Yang JJ, Metcalfe CD. Carbamazepine and its metabolites in wastewater and in
biosolids in a municipal wastewater treatment plant. Environ Sci Technol 2005;39:
7469–75.
Michael I, Vasquez MI, Hapeshi E, Haddad T, Baginska E, Kummerer K, et al. Metabolites
and transformation products of pharmaceuticals in the aquatic environment as
contaminants of emerging concern. In: Lambropoulou D, Nollet L, editors. Transfor-
mation products of emerging contaminants in the environment. UK: Wiley; 2014.
p. 413–58.
Miège C, Choubert JM, Ribeiro L, Eusèbe M, Coquery M. Fate of pharmaceuticals and per-
sonal care products in wastewater treatment plants — conception of a database and
first results. Environ Pollut 2009;157:1721–6.
Nelson ED, Do H, Lewis RS, Carr SA. Diurnal variability of pharmaceutical, personal care
product, estrogen and alkylphenol concentrations in effluent from a tertiary waste-
water treatment facility. Environ Sci Technol 2011;45:1228–34.
Nikolaou A, Meric S, Fatta D. Occurrence patterns of pharmaceuticals in water and waste-
water environments. Anal Bioanal Chem 2007;387:1225–34.
Ohlenbusch G, Kumke MU, Frimmel FH. Sorption of phenols to dissolved organic matter
investigated by solid phase microextraction. Sci Total Environ 2000;253:63–74.
Ollers S, Singer HP, Fassler P, Muller SR. Simultaneous quantification of neutral and acidic
pharmaceuticals and pesticides at low ng/L level in surface and wastewater. J
Chromatogr A 2001;911:225–34.
Olsen B, Munster VJ, Wallensten A, Waldenstrom J, Osterhaus A, Fouchier RAM. Global
patterns of influenza A virus in wild birds. Science 2006;312:384–8.
Ort C, Lawrence MG, Reungoat J, Eaglesham Geoff, Carter S, Keller J. Determining the frac-
tion of pharmaceutical residues in wastewater originating from a hospital. Water Res
2010;44:605–15.
Ortiz de García S, Pinto Pinto G, García Encina P, Irusta Mata R. Consumption and occur-
rence of pharmaceutical and personal care products in the aquatic environment in
Spain. Sci Total Environ 2013a;444:451–65.
Ortiz de García S, Pinto GP, García-Encina PA, Mata RI. Ranking of concern, based on en-
vironmental indexes, for pharmaceutical and personal care products: an application
to the Spanish case. J Environ Manage 2013b;129:384–97.
Pentikainen PJ, Neuvonen PJ, Penttila A. Pharmacokinetics of metformin after intravenous
and oral administration to man. Eur J Clin Pharmacol 1979;16:195–202.
Pérez S, Barceló D. Application of advanced MS techniques to analysis and identification of
human and microbial metabolites of pharmaceuticals in the aquatic environment.
Trends Anal Chem 2007;26:494–514.
Pérez S, Barceló D. First evidence for occurrence of hydroxylated human metabolites of
diclofenac and aceclofenac in wastewater using QqLIT–MS and QqTOF–MS. Anal
Chem 2008;80:8135–45.
Perez-Parada A, Aguera A, Gomez-Ramos MM, Garcνa-Reyes JF, Heinzen H, Fernandez-
Alba RR. Behavior of amoxicillin in wastewater and river water: identification of its
main transformation products by liquid chromatography/electrospray quadrupole
time-of-flight mass spectrometry. Rapid Commun Mass Spectrom 2011;25:731.
Petrie B, McAdam EJ, Lester JN, Cartmell E. Obtaining process mass balances of pharma-
ceuticals and triclosan to determine their fate during wastewater treatment. Sci
Total Environ 2014;497:553–60.
Pieper C, Risse D, Schmidt B, Braun B, Szewzyk U, Rotard W. Water Res 2010;44:4559.
Postigo C, de Alda MJL, Barceló D. Analysis of drugs of abuse and their human metabolites
in water by LC–MS2: a non-intrusive tool for drug abuse estimation at the community
level. TrAC Trends Anal Chem 2008;27:1053–69.
Postigo C, de Alda MJL, Barceló D. Drugs of abuse and their metabolites in the Ebro
river basin: occurrence in sewage and surface water, sewage treatment plants
removal efficiency, and collective drug usage estimation. Environ Int 2010;36:
75–84.
Prasse C, Schlusener M, Schulz R, Ternes TA. Antiviral drugs in wastewater and surface
waters: a new pharmaceutical class of environmental relevance? Environ. Sci Technol
2010;44:1728–35.
Radjenovic J, Perez S, Petrovic M, Barcelo D. Identification and structural characterization
of biodegradation products of atenolol and glibenclamide byliquid chromatography
coupled to hybrid quadrupole time-offlightand quadrupole ion trap mass spectrom-
etry. J Chromatogr 2008;1210:142–53.
 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
Reddy S, Iden CR, Brownawell B. Analysis of steroid conjugates in sewage influent and ef-
fluent by liquid chromatography–tandem mass spectrometry. Anal Chem 2005;77:
7032–8.
Reith DM, Appleton DB, Hooper W, Eadie MJ. The effect of body size on the metabolic
clearance of carbamazepine. Biopharm Drug Dispos 2000;21:103–11.
Roberts PH, Thomas KV. The occurrence of selected pharmaceuticals in wastewater efflu-
ent and surface waters of the lower Tyne catchment. Sci Total Environ 2006;356:
143–53.
Ronfeld RA, Tremaine LM, Wilner KD. Pharmacokinetics of sertraline and its N-demethyl
metabolite in elderly and young male and female volunteers. Clin Pharmacokinet
1997;32:22–30.
Rosal R, Rodríguez A, Perdigón-Melón JA, Petre A, García-Calvo E, Gómez MJ, et al. Occur-
rence of emerging pollutants in urban wastewater and their removal through biolog-
ical treatment followed by ozonation. Water Res 2010;44:578–88.
Roth HJ, Fenner H. Struktur-Bioreaktivit¨at-WirkungsbezogeneEigenschaften. 2nd ed.
Stuttgart: Georg Thieme Verlag; 1994. p. 65–8.
Routledge EJ, Parker J, Odum J, Ashby J, Sumpter JP. Some alkyl hydroxy benzoate preser-
vatives parabens; are estrogenic. Toxicol Appl Pharmacol 1998a;153:12–9.
Routledge EJ, Sheahan D, Desbrow C, Brighty GC, Waldock M, Sumpter JP. Identification of
Estrogenic Chemicals in STW Effluent. 2. In Vivo Responses in Trout and Roach. Envi-
ron Sci Technol 1998b;32:1559–65.
Rubirola A, Llorca M, Rodriguez-Mozaz S, Casas N, Rodriguez-Roda I, Barceló D, et al. Char-
acterization of metoprolol biodegradation and its transformation products generated
in activated sludge batch experiments and in full scale WWTPs. Water Res 2014;63:
21–32.
Rule KL, Ebbett VR, Vikesland PJ. Formation of chloroform and chlorinated organics by
free-chlorine-mediated oxidation of triclosan. Environ Sci Technol 2005;39:3176–85.
Salgado R, Marques R, Noronha JP, Mexia JT, Carvalho G, Oehmen A, et al. Assessing the
diurnal variability of pharmaceutical and personal care products in a full-scale acti-
vated sludge plant. Environ Pollut 2011;159:2359–67.
Salgado R, Marques R, Noronha JP, Carvalho G, Oehmen A, Reis MAM. Assessing the re-
moval of pharmaceuticals and personal care products in a full-scale activated sludge
plant. Environ Sci Pollut Res 2012;19:1818–27.
Sammartino MP, Bellanti F, Castrucci M, Ruiu D, Visco G, Zoccarato T. Ecopharmacology:
deliberated or casual dispersion of pharmaceutical principles, phytosanitary, personal
health care and veterinary products in environment needs a multivariate analysis or
expert systems for the control, the measure and the remediation. Microchem J 2008;
88:201–9.
Santos LHMLM, Araújo AN, Fachini A, Pena A, Delerue-Matos C, Montenegro MCBSM. Eco-
toxicological aspects related to the presence of pharmaceuticals in the aquatic envi-
ronment. J Hazard Mater 2010;175:45–95.
Santos LHMLM, Gros M, Rodriguez-Mozaz S, Delerue-Matos C, Pena A, Barceló D, et al.
Contribution of hospital effluents to the load of pharmaceuticals in urban wastewa-
ters: identification of ecologically relevant pharmaceuticals. Sci Total Environ 2013;
461–462:302–16.
Sawchuk RJ, Maloney JA, Cartier LL, Rackley RJ, Chan KKH, Lau HSL. Analysis of diclofenac
and four of its metabolites in human urine by HPLC. Pharm Res 1995;12:756–62.
Schlusener MP. Fate, pathways and methods for the determination of selected antibiotics
and steroid hormones in the environment [Doctor dissertation] University of
Duisburgessen; 2005.
Schmidt R, Brockmeyer R. Vom Wasser 2002;98:37–54.
Schneider W, Degen PH. Simultaneous determination of diclofenac sodium and its hy-
droxy metabolites by capillary column gas chromatography with electron-capture
detection. J Chromatogr A 1981;217:263–71.
Schrey P, Wilhelm M. Rückstände von Arzneimitteln in Wasserproben — Befunde und
deren Bewertung aus Sicht der Trinkwasserverordnung. Teil I. DVGW-Schr.reihe.
Wasser 1999;94:1–70.
Schroeder DH. Metabolism and kinetics of bupropion. J Clin Psychiatry 1983;15:79–781.
Schultz MM, Furlong ET. Trace analysis of antidepressant pharmaceuticals and their select
degradates in aquatic matrixes by LC/ESI/MS/MS. Anal Chem 2008;80:1756–61.
Schulz M, Löffler D, Wagner M, Ternes TA. Transformation of the X-ray contrast medium
iopromide in soil and biological wastewater treatment. Environ Sci Technol 2008;42:
7207–17.
Schutz H, Feldmann KF, Faigle JW, Kriemler HP, Winkler T. The metabolism of 14C-
oxcarbazepine in man. Xenobiotica 1986;16:769–78.
Schwabe U, Paffrath D. Arzneiverordnungs — report 2008. Berlin, Heidelberg: Springer;
2008.
Schymanski EL, Singer HP, Longrée P, Loos M, Ruff M, Stravs MA, et al. Strategies to char-
acterize polar organic contamination in wastewater: exploring the capability of high
resolution mass spectrometry. Environ Sci Technol 2014;48:1811–8.
Shackleton RHL. J Chromatogr 1986:91–156.
Shao B, Chen D, Zhang J, Wu Y, Sun C. Determination of 76 pharmaceutical drugs by liquid
chromatography–tandem mass spectrometry in slaughterhouse wastewater. J
Chromatogr A 2009;1216:8312–8.
Simons J. The $10 billion pill, Fortune magazine; 2003.
Singer AC, Howard BM, Johnson AC, Knowles CJ, Jackman S, Accinelli C, et al. Meeting re-
port: risk assessment of Tamiflu use under pandemic conditions. Environ Health
Perspect 2008;116:1563–7.
Soni MG, Carabin IG, Burdock GA. Safety assessment of esters of p-hydroxybenzoic acid
parabens. Food Chem Toxicol 2013;43:985–1015.
Soufan M, Deborde M, Delmont A, Legube B. Aqueous chlorination of carbamazepine:
kinetic study and transformation product identification. Water Res 2013;47:
5076–87.
Stamatis NK, Konstantinou IK. Occurrence and removal of emerging pharmaceutical, per-
sonal care compounds and caffeine tracer in municipal sewage treatment plant in
Western Greece. J Environ Sci Health B 2013;48:800–13.
925
Stülten D. Occurrence of diclofenac and selected metabolites in sewage effluents. Sci Total
Environ 2008;405:310–6.
Stülten D, Lamshöft M, Zühlke S, Spiteller M. Isolation and characterization of a new
human urinary metabolite of diclofenac applying LC–NMR–MS and high-resolution
mass analyses. J Pharm Biomed Anal 2008;47:371–6.
Stumpf M, Ternes TA, Haberer K, Baumann W. Isolierung von. Ibuprofen-Metaboliten und
deren edeutung als Kontaminanten der aquatischen Umwelt. Vom Wasser 1998;91:
291–303.
Suárez S, Carballa M, Omil F, Lema JM. How are pharmaceutical and personal care prod-
ucts PPCPs; removed from urban wastewaters? Rev Environ Sci Biotechnol 2008;7:
125–38.
Tanaka E, Honda K, Yasuhara H. Ketamine: its pharmacology and toxicology. Jpn J Foren
Toxicol 2005;23:187–91.
Terasaki M, Kamata R, Shiraishi F, Makino M. Evaluation of the estrogenic activity of
parabens and their chlorinated derivatives by using yeast two-hybrid assay and the
enzyme-linked immunosorbent assay. Environ Toxicol Chem 2009a;28:204–8.
Terasaki M, Makino M, Tatarazako N. Acute toxicity of parabens and their chlorinated by-
products with Daphnia magna and Vibrio fischeri bioassays. J Appl Toxicol 2009b;29:
242–7.
Ternes TA. Occurrence of drugs in German sewage treatment plants and rivers. Water Res
1998;12:3245–60.
Ternes T, Hirsch R. Occurrence and behavior of X-ray contrast media in sewage facilities
and the aquatic environment. Environ Sci Technol 2000;34:2741–8.
Ternes TA, Stumpf M, Mueller J, Haberer K, Wilken R-D, Servos M. Behavior and occur-
rence of estrogens in municipal sewage treatment plants—I. Investigations in
Germany, Canada and Brazil. Sci Total Environ 1999a;225:81–90.
Ternes TA, Kreckel P, Mueller J. Behaviour and occurrence of estrogens in municipal sew-
age treatment plants — II. Aerobic batch experiments with activated sludge. Total
Environ 1999b;225:91–9.
Ternes TA, Andersen H, Gilberg D, Bonerz M. Determination of estrogens in sludge and
sediments by liquid extraction and GC–MS/MS. Anal Chem 2002;74:3498–504.
Terzić S, Senta I, Ahel M, Gros M, Petrović M, Barcelo D, et al. Occurrence and fate of
emerging wastewater contaminants in Western Balkan Region. Sci Total Environ
2008;399:66–77.
Terzic S, Senta I, Ahel M. Illicit drugs in wastewater of the city of Zagreb Croatia; —
estimation of drug abuse in a transition country. Environ Pollut 2010;158:2686–93.
Thomas PM, Foster GD. Tracking acidic pharmaceuticals, caffeine, and triclosan through
the wastewater treatment process. Environ Toxicol Chem 2005;2:25–30.
Trautwein C, Kümmerer K. Incomplete degradation of the anti-diabetic Metformin and
identification of the microbial dead-end transformation product Guanylurea.
Chemosphere 2011;85:765–73.
Tyler CR, Jobling S, Sumpter JP. Crit Rev Toxicol 1998;28:319–61.
van Nuijs ALN, Pecceu B, Theunis L, Dubois N, Charlier C, Jorens PG, et al. Cocaine and me-
tabolites in waste and surface water across Belgium. Environ Pollut 2009;157:123–9.
van Nuijs ALN, Castiglioni S, Tarcomnicu I, Postigo C, de Alda ML, Neels H, et al. Illicit drugs
consumption estimations derived from wastewater analysis: a critical review. Sci
Total Environ 2011;409:3564–77.
Vanderford BJ, Snyder SA. Analysis of pharmaceuticals in water by isotope dilution liquid
chromatography/tandem mass spectrometry. Environ Sci Technol 2006;40:7312–20.
Vasskog T, Anderssen T, Pedersen-Bjergaard S, Kallenborn R, Jensen E. Occurrence of se-
lective serotonin reuptake inhibitors in sewage and receiving waters at Spitsbergen
and in Norway. J Chromatogr A 2008;1185:194–205.
Verenitch SS, Lowe CJ, Mazumder A. Determination of acidic drugs and caffeine in munic-
ipal wastewaters and receiving waters by gas chromatography–ion trap tandem
mass spectrometry. J Chromatogr A 2006;1116:193–203.
Verlicchi P, Al Aukidy M, Zambello E. Occurrence of pharmaceutical compounds in
urban wastewater: removal, mass load and environmental risk after a secondary
treatment — a review. Sci Total Environ 2012;429:123–55.
VICH. Environmental impact assessment (EIAs) for veterinary medicinal products
(VMPs)—phase I. http://www.vichsec.org/pdf/2000/Gl06_st7.pdf, . [accessed 11.02.
13].
Vieno NM, Tuhkanen T, Kronberg L. Seasonal variation in the occurrence of pharmaceuti-
cals in effluents from a sewage treatment plant and in the recipient water. Environ
Sci Technol 2005;39:8220–6.
Vieno NM, Tuhkanen T, Kronberg L. Analysis of neutral and basic pharmaceuticals in sew-
age treatment plants and in recipient rivers using solid phase extraction and liquid
chromatography–tandem mass spectrometry detection. J Chromatogr A 2006;1134:
101–11.
von Bruchhausen F, Dannhardt G, Ebel S, Frahm AW, Holzgrabe U. Hagers Handbuch der
Pharmazeutischen Praxis. , 5th ed.Berlin: Springer-Verlag; 1994.
Warrington SJ, Ronfeld RA, Wilner KD, Lazar JD. Human pharmacokinetics of sertraline.
Clin Neuropharmacol 1992;15:54–5.
Weigel S, Berger U, Jensen E, Kallenborn R, Thoresen H, H€uhnerfuss H. Determination of
selected pharmaceuticals and caffeine in sewage and seawater from Tromsø/Norway
with emphasis on ibuprofen and its metabolites. Chemosphere 2004;56:583–92.
Weissbrodt D, Kovalova L, Ort C, Pazhepurackel V, Moser R, Hollender J, et al. Mass flows
of X-ray contrast media and cytostatics in hospital wastewater. Environ Sci Technol
2009;43:810–4817.
Wick A, Wagner M, Ternes TA. Elucidation of the transformation pathway of the opium
alkaloid codeine in biological wastewater treatment. Environ Sci Technol 2011;45:
3374–85.
Writer JH, Ferrer I, Barber LB, Thurman EM. Widespread occurrence of neuro-active phar-
maceuticals and metabolites in 24 Minnesota rivers and wastewaters. Sci Total Envi-
ron 2013;461–462:519–27.
Xiao XY, McCalley DV, McEvoy J. Analysis of estrogens in river water and effluents using
solid-phase extraction and gas chromatography–negative chemical ionization mass
926 E.N. Evgenidou et al. / Science of the Total Environment 505 (2015) 905–926
spectrometry of the pentafluorobenzoyl derivatives. J Chromatogr A 2001;923:
195–204.
Xu W, Zhang G, Li X, Zouc S, Lia P, Hua Z, et al. Occurrence and elimination of antibiotics at
four sewage treatment plants in the Pearl River Delta PRD; South China. Water Res
2007;41:4526–34.
Zonja B, Aceña J, Jelic A, Petrovic M, Perez S, Barceló D. Transformation products of emerg-
ing contaminants: analytical challenges and future needs, In. In: Lambropoulou DA,
Nollet LML, editors. Transformation products of emerging contaminants in the envi-
ronment: analysis, processes, occurrence, effects and risks. 1st ed. John Wiley and
Sons Ltd; 2014. p. 303–20.
Zuccato E, Castiglioni S. Illicit drugs in the environment. Philos Trans A Math Phys Eng Sci
2009;367:3965–78.
Zuccato E, Chiabrando C, Castiglioni S, Calamari D, Bagnati R, Schiarea S, et al. Cocaine in
surface water: a new evidence-based tool to monitor community drug abuse. Environ
Health Glob 2005;4:14–20.
Zuccato E, Chiabrando C, Castiglioni S, Bagnati R, Fanelli R. Estimating community drug
abuse by wastewater analysis. Environ Health Perspect 2008;116:1027–32.
Zuehlke S, Duennbier U, Heberer T. Determination of polar drug residues in sewage and
surface water applying liquid chromatography–tandem mass spectrometry. Anal
Chem 2004;76:6548–54.
Zuehlke S, Duennbier U, Heberer T. Investigation of the behavior and metabolism of phar-
maceutical residues during purification of contaminated ground water used for
drinking water supply. Chemosphere 2007;69:1673–80.
Zwiener C, Seeger S, Glauner T, Frimmel FH. Metabolites from biodegradation of pharma-
ceutical residues of ibuprofen in biofilm reactors and batch experiments. Anal Bioanal
Chem 2002;372:569–75.