7th International Workshop on Biomonitoring of Atmospheric Pollution (BIOMAP)
June 14-19, 2015
Lisbon, Portugal
Determination of Estrogenic Activity in PM10
Air Samples from Flanders (Belgium) with the
BG1 Ovarian Cancer Cell Line
K. CROES1, R. VAN DEN HEUVEL2, E. DEN HOND2, M. PLUSQUIN3, J.
STAELENS4, B. VAN DEN BRIL4, M.S. DENISON5, T. VANDERMARKEN1, K.
VAN LANGENHOVEN1, M. ELSKENS1
1
Department of Analytical, Environmental and Geo-Chemistry (AMGC), Vrije Universiteit Brussel;
Pleinlaan 2, 1050 Brussels, Belgium; kim.croes@vub.ac.be, tara.vandermarken@vub.ac.be,
kavlange@vub.ac.be, melskens@vub.ac.be
2
Environmental Health and Risk, VITO; Boeretang 200, 2400 Mol, Belgium;
rosette.vandenheuvel@vito.be, elly.denhond@vito.be
3
Interuniversity Institute for Biostatistics and Statistical Bioinformatics, Hasselt University; Agoralaan
1, 3590 Diepenbeek, Belgium; michelle.plusquin@uhasselt.be
4
Flemish Environment Agency (VMM); Kronenburgstraat 45, 2000 Antwerp, Belgium;
j.staelens@vmm.be
5
Department of Environmental Toxicology, University of California; Davis, CA 95616 USA;
msdenison@ucdavis.edu
Exogenous substances that act like hormones and disrupt the physiologic function of endogenous
hormones in the endocrine system are called endocrine disrupting compounds (EDCs). Due to their
possible health effects, there is growing attention for EDC pollution in our environment. In this study, a
novel methodology to measure the overall response of estrogenic disruptors in PM10 air samples was
developed and the effect of different extraction solvents was investigated. To obtain more information
about the EDCs present in our environment and their effects on the human hormone system, a bioassay
technique using a BG1Luc4E2 ovarian carcinoma cell line was employed to quantify the potency of the
mixture of estrogenic compounds in PM10 air samples. Between April 2013 and January 2014, samples
were taken in a rural, industrial and urban area in Flanders (Belgium). Estrogen disruptive potencies
were reported as biologic equivalents (BEQ), expressed in picograms 17β-estradiol per cubic meter of
air. Non-parametric tests (Kruskal-Wallis and Wilcoxon rank tests) were used to determine possible
differences between sampling areas and time periods. Spearman’s rank correlations were calculated to
examine relationships with meteorological data and environmental pollutants. No statistical significant
differences in estrogenic activity were found between the locations, but large day-by-day variations
were seen in all locations. The estrogenic activity in the industrial area was positively related with the
sum of pyrene and fluoranthene, while a borderline significant negative correlation with air temperature
and sum of the carcinogenic PAHs was found. Furthermore, the estrogenic potency was generally
higher when a polar extraction solvent was used (ethanol or acetonitrile) instead of a mixture of a polar
and non-polar solvent (hexane/acetone, 50/50). This study shows that this in vitro bioassay is a suitable
tool to monitor estrogenic disruption in air samples. Since relatively large day-by-day fluctuations were
observed, it seems that on all locations a certain background estrogenic activity is present with on
certain occasions high peak levels originating from a mixture of estrogen-active compounds. More long
term studies on PM10, or even PM2.5 and PM1, are thus needed to investigate these fluctuations in
estrogenic potency in relation to meteorological and environmental data and to define possible human
health effects. However, it seems that changes in sample pre-treatment methods can significantly
influence the observed potencies and that a harmonized approach for sampling, extraction and in vitro
measurement is thus necessary.
Keywords: PM10, CALUX, EDC, estrogen, bioassay