Regulatory Toxicology and Pharmacology 55 (2009) 367371

Contents lists available at ScienceDirect

Regulatory Toxicology and Pharmacology

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

WikiPharma A freely available, easily accessible, interactive and

comprehensive database for environmental effect data for pharmaceuticals
Linda Molander, Marlene gerstrand *, Christina Rudn
Royal Institute of Technology/Philosophy, Teknikringen 78B, 10044 Stockholm, Sweden

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

Article history: A signicant number of Active Pharmaceutical Ingredients (APIs) have been identied in the environment
Received 23 June 2009 and in surface waters. Data on the environmental hazards associated with these substances are emerging
Available online 29 August 2009 but are still scarce. We have compiled publicly available ecotoxicity data for APIs into a database called
WikiPharma. The use of the database is free of charge. It can be accessed and updated continuously as a
Keywords: wiki. The aim of WikiPharma is to provide an easily accessible, comprehensive and up-to-date overview
Pharmaceuticals of effects caused by pharmaceuticals on non-target organisms. The database currently contains basic
Ecotoxicity
information, i.e. substance name, ATC code(s) and pharmaceutical group(s), for 831 APIs representing
Database
Environmental effects
35 different drug classes. Effect data have been identied and included for 116 of these substances. These
MistraPharma ecotoxicity test data have been extracted from 156 different sources. The development of a comprehen-
WikiPharma sive database on ecological hazard of APIs can facilitate identication of data gaps and promote environ-
mental risk assessment of these substances. The database is available at www.wikipharma.org.
2009 Elsevier Inc. All rights reserved.

1. Introduction The aim of the WikiPharma is to provide an easily accessible

Up to now almost 200 pharmaceutical substances have been
identied in surface waters. The occurrence of Active Pharmaceu-
tical Ingredients (APIs) in the aquatic environment is certainly a
cause for concern, but exposure is only one prerequisite for there
being a risk to the environment. Evidence of harm is the other.
So far, relatively little is known about the risks associated with
the occurrence of these compounds in the environment but given
their potency to interact with biological target molecules that in
many cases are highly conserved among a variety of organisms
(Gunnarsson et al., 2008), a risk to non-target species cannot easily
be dismissed.
To identify information gaps and thereby contributing to high-
lighting research needs, we have compiled publicly available eco-
toxicity data into a database using Microsoft Access 2000
software. The database is called WikiPharma. It is available at no
cost and constructed as a wiki so that all users can propose data
to be added. This solution enables continuous updating of the data-
base. Only published data are allowed to be entered in the data-
base. The peer-review system for published data is used as a
quality control. WikiPharma provides extracted data and biblio-
graphic references to these data. It does however not give direct ac-
cess to the papers from where the data is gathered.
* Corresponding author. Fax: +46 8 790 95 99.
E-mail address: maa2@kth.se (M. gerstrand).
overview of effects caused by pharmaceuticals on non-target
organisms identied in acute, sub-chronic and chronic ecotoxicity
tests. This concentration of ecotoxicity data for pharmaceuticals in
one single place can be of help when assessing potential risks of
pharmaceutical ingredients in the environment, as well as for iden-
tifying data gaps.
This paper presents the database construction, its contents and
guidance for users on how to search for data. Furthermore,
quantitative characteristics of the database are presented, such as
the frequency of different types of test methods, test species and
effect measures as well as what pharmaceutical substances
seem to have the highest toxicity. The database is available at
www.wikipharma.org.
The database was developed within the Swedish research pro-
gramme MistraPharma (www.mistrapharma.se). MistraPharma
concerns the identication and reduction of environmental risks
caused by the use of human pharmaceuticals and is funded by
the Swedish Foundation for Strategic Environmental Research
(Mistra).
2. Scope of the database
The database intends to include all available ecotoxicity data
for APIs. However, in the current version of the database only
the APIs for which an environmental risk assessment is required
according to European law are included, hence vitamins, miner-
als, vaccines, etc. are not. The reason for the exceptions is that

0273-2300/$ - see front matter 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.yrtph.2009.08.009
368 L. Molander et al. / Regulatory Toxicology and Pharmacology 55 (2009) 367371
these APIs are generally considered to be of low environmental
risk.
The following criteria were initially used to select data to be in-
cluded in the database:
 The data should be published in a peer reviewed journal and
presented in the English language.
 Both in vivo and in vitro tests on non-target species were
included.
 Only tests where an EC50, IC50, LC50, NOEC, LOEC, NOEL, LOEL,
NOAEL or LOAEL is presented, or for which such effect measures
can be derived or calculated, were included.
 The focus is on reports of adverse effects, and also on biological
alterations that might lead to an adverse effect. Data on occur-
rence, fate and exposures (including bioconcentration) were
not included.
When available, the following information has been included in
the database:
 Complete bibliographic reference.
 The name of the tested API.
 The type of test method used (e.g. OECD or ASTM standard
methods).
 The identity of the tested species.
 The sex of the tested organisms (when relevant).
 The age and/or life stage of the tested organisms.
 Number of organisms used in the test.
 Concentrations/doses of the API used in the test (nominal and/or
measured).
 Exposure time.
 Study period (if different from exposure time).
 Route of exposure (if other than through water exposure).
 The result of the test reported as one of the following effect
measures: EC50, IC50, LC50, NOEC, LOEC, NOEL, LOEL (NOAEL
and LOAEL were reported as NOEL and LOEL, respectively).
 The critical effect (endpoint) identied in the test.
 Any additional information or comments about the test or pub-
lication that could be useful.
3. Identication of data search strategy
Ecotoxicity data were systematically searched for in relevant
scientic databases such as CSA, ScienceDirect and PubMed. CSA
includes 17 biology databases: ASFA 3 Aquatic Pollution and
Environmental Quality, Bacteriology Abstracts (Microbiology B),
BioOne Abstracts and Indexes, Biotechnology Research Abstracts,
Conference Papers Index, Ecology Abstracts, EIS Digests of Envi-
ronmental Impact Statements, Environmental Engineering
Abstracts, TOXLINE, Health and Safety Science Abstracts, Human
Population & Natural Resource Management, Industrial and
Applied Microbiology Abstracts (Microbiology A), Pollution
Abstracts, Risk Abstracts, Sustainability Science Abstracts, Toxicol-
ogy Abstracts, Water Resources Abstracts. The search engine Goo-
gle Scholar was also a useful tool. In addition to these search paths
data from relevant books were extracted, and the reference lists of
the retrieved articles including review articles were searched for
information about additional publications.
When searching for data on these substances a number of key-
words were used to obtain relevant information (Table 1). Since
the MistraPharma research programme does not cover the terres-
trial compartment, the main focus was on effect studies on non-
target organisms living in the aquatic environment. Terrestrial
studies were however included when identied.
The aim of the search procedures was to identify all the data
that fullled our initial criteria and we believe that this search
Table 1
Keywords used when searching for information in databases.
Keywords
Name of API
Name of API and ecotoxic*
Name of API and environment*
Name of API and environment* and effect*
Name of API and environment* and toxic*
Name of API and EC50 or LC50 or NOEC or LOEC
Pharmaceutical* and environment*
Pharmaceutical* and aquatic and environment*
Pharmaceutical* and aquatic and toxic*
Pharmaceutical* and EC50 or LC50 or NOEC or LOEC
*
Word stem search.
strategy resulted in a good coverage of the publicly accessible eco-
toxicity data.
4. Database structure
The database is structured according to standard Microsoft Ac-
cess principles using tables and established relations between
these tables. There are three tables managing the identity of the
substance and the function of the API, i.e. substance name, ATC-
code(s)1 and pharmaceutical group(s).
The other two tables contain the ecotoxicity data and the corre-
sponding bibliographic references, respectively. The overall struc-
ture of the database is shown in Fig. 1.
5. Searching the database
The search engine is constructed so that a specic pharmaceuti-
cal ingredient or a whole drug class, e.g. non-steroidal anti-inam-
matory drugs (NSAIDs), can be selected from a drop-down-list to
obtain all effect data of interest. Information from the WikiPharma
database can also be retrieved by downloading all information in
the database in one table or by downloading the whole database
(requires Microsoft Access). The format of the data extracted from
a Microsoft Access database is fully compatible with other Ofce
software such as Excel and Word.
6. Overview of database contents
In total the database contains basic information, i.e. substance
name, ATC code(s) and pharmaceutical group(s), for 831 APIs rep-
resenting 35 different pharmaceutical drug classes. Effect data
have been found in the publicly available literature for 116 of these
substances. These ecotoxicity test data have been extracted from
156 different sources.
Approximately 60% of the included effect data originates from
tests conducted according to standardized methods. The most fre-
quently occurring testing standards are those issued by OECD, the
U.S. EPA, and ISO. The mainly used test guidelines are presented in
Table 2. Modied versions of the standard test guidelines were
included in the counting.
1
Every API has an ATC code according to the Anatomical Therapeutic Chemical
(ATC) classication system established by the World Health Organization (WHO). In
the ATC classication system the substances are categorized into groups depending
on which organ or system they are designed to act on and according to their chemical,
pharmacological and therapeutic properties. In cases when an API has more than one
area of use there is also more than one ATC code for this ingredient (WHO, 2008). To
enable searches on all ATC codes in the database a table was created linking one of the
ATC codes to all codes for that particular substance.
 L. Molander et al. / Regulatory Toxicology and Pharmacology 55 (2009) 367371 369

Fig. 1. The database structure.

Tests performed using standard kits like the Microtox test, Table 3
Thamnotoxkit, Rotoxkit, Spirotox, Streptoxkit and Artoxkit etc. Top 10 used test species in ecotoxicity testing of pharmaceutical substances.

are also commonly occurring in the database.
There are 127 different species represented in the data set.
Since the majority of the tests compiled in the database were per-
formed according to standardized test methods, most of the test
organisms used is consequently also standard test species. The
top 10 most frequently used test species are listed in Table 3. Both
in vivo and in vitro tests were included in the counting of tests with
different species.
Crustacean species (Daphnia magna, Ceriodaphnia dubia domi-
nate) are the most commonly used species when looking at the
whole data set. This is not very surprising since daphnid species
are extensively used in freshwater toxicity testing because they
are abundant and widely distributed, easy to maintain in the
laboratory and are sensitive towards a broad range of environ-
mental contaminants (Cooney, 2003). The marine bacterium Vib-
rio sheri is also recommended as test species and commonly
used for ecotoxicity testing of antibiotic agents (Backhaus and
Grimme, 1999). Three different sh species are among the top
ten, however, the majority of the tests with Poeciliopsis lucida
and Oncorhynchus mykiss were conducted in vitro. Ecotoxicity
testing with sh are often used to determine adverse effects of
the estrogens 17-bestradiol and 17a-ethinylestradiol, where
vitellogenin (VTG) induction is considered a valuable biomarker
for assessing exposure to environmental estrogens in sh (e.g.
Tyler et al., 2002).
More than half of the compiled data represent short-term expo-
sure and acute responses. An estimation of the number of acute
Table 2
Most frequently used test standards in ecotoxicity testing of pharmaceutical
substances.
Standard/guideline Number of tests (%)
OECD 225 (33.2)
USEPA 171 (25.2)
ISO 141 (20.8)
ASTM 76 (11.2)
EEC 37 (5.5)
AFNOR 28 (4.1)
OECD, Organization for Economic Co-operation and Development; ASTM, American
Society for Testing and Materials; EPA, Environmental Protection Agency (US); ISO,
International Standards Organization; EEC, European Economic Community;
AFNOR, Association Franaise de Normalisation.
Species Number of tests (%)
Daphnia magna (crustacean) 250 (29.1)
Vibrio scheri (bacterium) 104 (12.1)
Pseudokirchneriella subcapitata (algae) 82 (9.5)
Poeciliopsis lucida (sh)* 75 (8.7)
Oncorhynchus mykiss (sh)** 72 (8.4)
Glomus intraradices (mycorrhizal fungus) 60 (7.0)
Ceriodaphnia dubia (crustacean) 59 (6.9)
Pimephales promelas (sh) 57 (6.6)
Brachionus calyciorus (rotifer) 53 (6.2)
Thamnocephalus platyurus (crustacean) 47 (5.5)
*
In vitro test with P. lucida hepatocytes in 70 tests.
**
In vitro test with O. mykiss hepatocytes in 48 tests.
and chronic ecotoxicity tests gathered in the database was ob-
tained by grouping different species together due to generation
time. Based on the rough division of species into microorganisms
(mainly bacteria), algae (including vascular plants), invertebrates
and vertebrates (sh and amphibians) tests classied as acute
(i.e. exposure 630 min for microorganisms, 672 h for algae,
648 h for invertebrates, 696 h for vertebrates)2 constitute 55% of
the WikiPharma data set.
In addition, the total number of the effect measures EC50, LC50
and IC50 exceed the number of NOEC and LOEC values (Table 4),
which could be seen as an indicator of a higher frequency of acute
tests. Although, median effect concentrations are often obtained
from short-term toxicity tests it is worth noting that such effect
measures can also result from tests where the exposure is consid-
ered to be sub-chronic or chronic (Jop, 1997). Thus, this is only an
estimation of the amount of short-term and long-term tests com-
piled in the database.
Data from ecotoxicity testing compiled in the database show
that the natural estrogen 17b-estradiol and the synthetic estrogen
17a-ethinylestradiol are the substances for which the lowest effect
concentrations have been reported. Both the lowest short-term
2
Exposure time classied as acute was done according to Cooney (2003) for
invertebrate and vertebrate species. For algal species, 72 h E(L)C50 values were
classied as acute effect concentrations while 72 h NOEC and LOEC values were
classied as chronic effect concentrations in accordance with the Technical Guidance
Document (TGD) on Risk Assessment (TGD, 2003). Exposure time for microorganisms
was either 30 min or 20 h. The rst was regarded as acute, the latter as chronic.
370 L. Molander et al. / Regulatory Toxicology and Pharmacology 55 (2009) 367371
Table 4
Frequency of different effect measures used in ecotoxicity testing of pharmaceutical
substances.
Effect measure Number of effect measures (%)
EC50 852 (45.8)
LC50 232 (12.8)
IC50 86 (4.6)
NOEC 240 (12.9)
LOEC 450 (24.2)
median effect concentrations (EC50s) and long-term NOEC and
LOEC values are found for these estrogens. Among the lower NOEC
and LOEC values, ranging from 0.4 ng/L and from 0.03 ng/L respec-
tively up to a few micrograms per liter, a few effect concentrations
for the non-steroidal anti-inammatory drugs (NSAIDs) ibuprofen
and diclofenac and the selective serotonin reuptake inhibitor (SSRI)
uoxetine are found as well. Approximately 90 days of exposure of
17a-ethinylestradiol (EE2) at a concentration as low as 0.03 ng/L
signicantly induced intersex in the Japanese ricesh Oryzias latipes
(Metcalfe et al., 2001). This is an environmental relevant concentra-
tion since EE2 concentrations up to 7 ng/L have been detected in
European sewage treatment plant efuents and generally ranges
between 0.5 and less than 0.1 ng/L in surface waters (Desbrow
et al. 1998; Lnge et al., 2001).
Another pharmaceutical group identied in the database for
producing low median effect concentrations is the antibiotics, rep-
resented by amoxicillin, levooxacin, ooxacin and ciprooxacin.
The lowest EC50 values were identied at a few lg/L.
Pharmaceutical concentrations in surface waters, groundwater and
sediment normally range from nanograms to a few micrograms per li-
ter (Kmmerer, 2004) and for the majority of the pharmaceutical ingre-
dients adverse effects have been identied at concentrations far above
environmentally measured concentrations.
7. Database availability, guidance and explanations
In some scientic articles no effect measures are presented
although signicant effects were seen in the study. Where appro-
priate, such critical effect concentrations have been interpreted
as for example a LOEC value. The basis for the interpretations is
the denition of a LOEC value as the lowest observed signicant ef-
fect concentration and a NOEC value as the highest concentration
where no signicant effect was observed on the test organisms
compared to the control in a specic test (Rand et al., 2003).
Whereas signicant effect concentrations have been labeled LOEC
when the test organisms were exposed to the drug substance via
the surrounding media, LOEL (Lowest Observed Effect Level) was
used in cases when the test organisms were injected with the sub-
stance. To inform the users searching information in the database
that this is our own interpretation a note about this has been made
in a column named Comment (Fig. 1).
In the comment column there is also room for making notes on
if something in the article is unclear or if there are more data avail-
able from a study that for some reason is not included in the data-
base. For example, in a study conducted by Marques et al. (2004)
containing very much effect data, vague effect data presented as
>10.00 mg/L were left out with the comment More effect data
on neonates per brood and age at each reproduction available for
which all NOEC and LOEC values >10.00 mg/L. Only the more
specied data were extracted.
One column in the APIData table is named Route of exposure to
which information regarding the route only was included if the test
organisms were exposed in any other way than via the surround-
ing test media, e.g. by injection.
In some cases information of both exposure time and study per-
iod can be found. While the exposure time describes the time a test
organism are exposed to a pharmaceutical ingredient, e.g. 48 h, the
study period can include more than one exposure period and/or
also include a recovery period after the exposure has ended.
There is a column in the APIData table named Substance com-
ment with the purpose of informing the users if the pharmaceutical
ingredient tested is a metabolite of a substance, e.g. salicylic acid is
the main metabolite of acetylsalicylic acid. In a few cases effect
data of metabolites of APIs were added to the database although
the name of the metabolite was unknown or it had not been given
any ATC code. In these cases the ATC code of the parent substance
was used and a comment that the test substance is a metabolite
was made. Comments of what compounds were added to the test
solutions, e.g. verapamil-HCl or diclofenac-Na, were also made in
this eld.
8. Turning the database into a Wiki
This paper describes the background and design of the database
as it is today. We now make the database available to the scientic
community and make it possible for anyone, after a simple login-
registration, to propose changes and additions to it. All proposed
changes to the database will have to be approved by a database
administrator. The administrator checks whether the proposed
additions seem reasonable and compatible with the aims of the
database. Data that have either been published in the open scien-
tic literature or that have been sufciently peer reviewed and
made publicly available by other means, will be included. Apart
from that there will be no additional quality evaluation of data be-
fore they are included and published in the database. In a wiki all
users and contributors are responsible for quality assurance. For
scrutiny and evaluation of the data, the user is referred to the full
reference.
The goal is that the WikiPharma should be an easy accessible,
comprehensive, and up-to-date source for information about the
environmental toxicity of APIs. If this goal will be reached is up
to the scientic community.
Conicts of interest
The authors declare that there are no conicts of interest.
Acknowledgments
The authors thank Anna stensson for help with the gathering
of data and Katarzyna Malkiewicz and three anonymous reviewers
for valuable comments on the paper. The study was funded by Mis-
tra, the Foundation for Strategic Environmental Research.
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