First Determination of UV Filters in Marine Mammals. Octocrylene Levels in Franciscana Dolphins

Abstract

Most current bioexposure assessments for UV filters focus on contaminants concentrations in fish from river and lake. To date there is not information available on the occurrence of UV filters in marine mammals. This is the first study to investigate the presence of sunscreen agents in tissue liver of Franciscana dolphin (Pontoporia blainvillei), a species under special measures for conservation. Fifty six liver tissue samples were taken from dead individuals accidentally caught or found stranded along the Brazilian coastal area (six States). The extensively used octocrylene (2-ethylhexyl-2-cyano-3,3-diphenyl-2-propenoate, OCT) was frequently found in the samples investigated (21 out of 56) at concentrations in the range 89-782 ng.g-1 lipid weight. São Paulo was found to be the most polluted area (70% frequency of detection). Nevertheless, the highest concentration was observed in the dolphins from Rio Grande do Sul (42% frequency of detection within that area). These findings constitute the first data reported on the occurrence of UV filters in marine mammals worldwide.

Full text

First Determination of UV Filters in Marine Mammals. Octocrylene
Levels in Franciscana Dolphins
Pablo Gago-Ferrero,
†
Mariana B. Alonso,
‡,§,∥
Carolina P. Bertozzi,
‡
Juliana Marigo,
‡
Lupércio Barbosa,
⊥
Marta Cremer,
#
Eduardo R. Secchi,
▽
Alexandre Azevedo,
∥
JoséLailson-Brito Jr.,
∥
Joao P. M. Torres,
§
Olaf Malm,
§
Ethel Eljarrat,
†
M. Silvia Díaz-Cruz,*
,†
and DamiàBarceló
†,○
†
Department of Environmental Chemistry, Water and Soil Quality Research Group, IDAEA-CSIC, Jordi Girona 18-26, 08034
Barcelona, Spain
‡
Projeto BioPesca, R. Paraguai, 241. Praia Grande, SP, Brasil, 11702-070.
§
Laboratory of Radioisotopes - Biophysics Institute (UFRJ), Av. Carlos Chagas Filho, 373 CCS - Bl. G, Rio de Janeiro, RJ, Brasil,
21941-902
∥
Laboratory of Aquatic Mammals and Bioindicators (UERJ), R. São Francisco Xavier, 524 - S.4018 - Bl. E, Rio de Janeiro, RJ, Brasil,
20550-013
⊥
Instituto ORCA, Vila Velha, ES, Brasil
#
Universidade de Joinvile (Univille), Joinvile, SC, Brasil
▽
Laboratory of Turtles and Marine Mammals (FURG), Rio Grande, RS, Brasil
○
Catalan Institute for Water Research (ICRA), Scientific and Technological Parc of the University of Girona, Emili Grahit, 101 Edifici
H2O, 17003 Girona, Spain
ABSTRACT: Most current bioexposure assessments for UV
filters focus on contaminants concentrations in fish from river
and lake. To date there is not information available on the
occurrence of UV filters in marine mammals. This is the first
study to investigate the presence of sunscreen agents in tissue
liver of Franciscana dolphin (Pontoporia blainvillei), a species
under special measures for conservation. Fifty six liver tissue
samples were taken from dead individuals accidentally caught
or found stranded along the Brazilian coastal area (six states).
The extensively used octocrylene (2-ethylhexyl-2-cyano-3,3-
diphenyl-2-propenoate, OCT) was frequently found in the
samples investigated (21 out of 56) at concentrations in the
range 89−782 ng·g−1lipid weight. São Paulo was found to be
the most polluted area (70% frequency of detection). Nevertheless, the highest concentration was observed in the dolphins from
Rio Grande do Sul (42% frequency of detection within that area). These findings constitute the first data reported on the
occurrence of UV filters in marine mammals worldwide.
■INTRODUCTION
UV filters (UV F) are emerging environmental contaminants
for which there is currently a lack of knowledge about their
occurrence, fate, and effects on the ecosystems.
1
UV F
constitutes a large and heterogeneous group of chemicals that
are ingredients in personal care products to protect skin and
hair from the sunlight, and in other industrial goods such as
paint, wax, plastic, or textile to prevent photodegradation of
polymers and pigments.
2
These chemicals enter the aquatic environment either
indirectly, via wastewater treatment plant effluents (urban and
industrial) or directly, through human aquatic recreational
activities. Previous studies have demonstrated the occurrence of
UV F in water, sewage sludge, sediment, and biota.
3−6
Many
UV F are lipophilic compounds, therefore have the potential for
bioaccumulation and biomagnification in aquatic ecosystems
through the trophic chain.
3
Works on biota were mainly
focused on fish,
2,3,7,8
but other organisms have been studied as
well, such as fish eating birds and aquatic invertebrates.
3
Several
UV filters are known to have toxic effects on both aquatic and
terrestrial organisms. Although the studies dealing with
ecotoxicity of these compounds is scarce, they have been
shown to act as environmental estrogens and antiandrogens,
cause reproductive disruption and affect the thyroid axis.
9,10
So
far, there is still even more limited information available about
Received: February 14, 2013
Revised: April 19, 2013
Accepted: April 29, 2013
Published: April 29, 2013
Article
pubs.acs.org/est
© 2013 American Chemical Society 5619 dx.doi.org/10.1021/es400675y |Environ. Sci. Technol. 2013, 47, 5619−5625

the fate and effects of these chemicals in marine ecosystems.
High levels of multiclass UV F in seawater have been reported,
with concentrations up to 799 ngL−1of 4-methylbenzylidene
camphor (4MBC).
11,12
Recently, it has been documented that
UV F caused harmful effects on coral reefs (coral bleaching) by
promoting viral infections.
13
As regards marine biota, the
analysis of four benzotriazole UV stabilizers, namely UV-320,
UV-326, UV-327, and UV-328, and the UV filter 4MBC in
marine organisms from the Ariake Sea (Japan) revealed that the
three benzotriazole stabilizers investigated bioaccumulated in all
the species analyzed, from benthic invertebrates to several fish
species, including the hammerhead shark.
14
Among UV filters, octocrylene (2-ethylhexyl-2-cyano-3,3-
diphenyl-2-propenoate, OCT) is of great concern since it is a
highly lipophilic compound (log Kow 6.88), stable, and resistant
to sunlight degradation, but there is evidence that it can trigger
the production of potentially harmful free radicals (reactive
oxygen species) when it releases the absorbed energy. The
widespread occurrence of this compound, as well as its high
concentrations found in sewage sludge and sediments
4,5
appears to be associated with its extensive use in formulations,
especially personal care products, because both protects in
UVA and UVB regions, and augments the absorbing capacity of
other organic UV filters, such as ethylhexylmethoxycinnamate
(EHMC), avobenzone (AVB), and benzophenone-3 (BP3).
15
Since maintaining the absorption capacity is important to
prevent erythema and to reduce the subsequent risk of
melanoma development, formulations containing OCT had
superior performance compared to other formulations that did
not contain OCT, and therefore, preferably used.
The goal of the present study was to contribute for a better
understanding of the impact of the increasing use of UV filters
in densely populated coastal areas on marine organisms. The
study aimed at demonstrating the potential for biomagnification
of the extensively used sunscreen agent OCT on marine
mammals, specifically on dolphin, since they occupy a higher
trophic level in the marine food chain, and have relatively low
metabolic activity, thus accumulating high levels of organic
pollutants in their body.
16
For this study Franciscana dolphin
(Pontoporia blainvillei) was the selected species. It is a small
cetacean with a distribution restricted to the southwest Atlantic
Ocean. This is the most impacted cetacean offthe eastern coast
of South America
17
and is listed as “vulnerable”in the Red
Book of the International Union for Conservation of the
Nature (IUCN). Franciscana was considered a species that
needs particularly measures of conservation
18
and is also
included in the Index II of the Convention on International
Trade in Endangered Species of Wild Fauna and Flora, that
Argentina, Uruguay, and Brazil are undersigned. Their coastal
distribution makes it particularly vulnerable to human activities
such as incidental capture in fisheries and habitat degradation
by anthropic contaminants.
19−21
Evidences suggest that the
mortality rates are excessive and unsustainable.
22
■EXPERIMENTAL SECTION
Chemicals and Reagents. OCT (>98% purity), and the
isotopically labeled compound benzophenone-d10 (BP-d10 99%
purity), used as internal standard (IS), were obtained from
Sigma-Aldrich (Steinheim, Germany). Organic solvents and
HPLC grade water (Lichrosolv), as well as H2SO4, formic acid
(98% purity) and hydromatrix were provided by Merck
(Darmstad, Germany). Nitrogen and argon (purchased from
Air Liquid, Barcelona, Spain) were of 99.995% purity. The
syringe and the pressurized liquid extraction (PLE) cellulose
filters used were purchased from Whatman (London, U.K.) and
from Dionex Corporation (Sunnyvale, CA), respectively.
Isolute Alumina Cartridges used for solid phase extraction
(SPE) were obtained from Biotage (Uppsala, Sweden).
The OCT and BP-d10 stock standard solutions were prepared
in methanol at 200 mg L−1. The solutions were stored in the
dark at −20 °C. A diluted 20 mg L−1stock standard solution
was prepared weekly. Working solutions were prepared daily by
appropriate dilution of the diluted stock standard solution.
Sampling Area and Sample Collection. The Brazilian
coastline has around 8500 km of length. The Southeast
Brazilian region, historically, had turned into an important
industrial center of Brazil. Rio de Janeiro and São Paulo States
are the most anthropogenically disturbed areas along the
country shoreline. Massive metropolitan complex surrounds the
estuaries and bays, which have been receiving discharges of
chemical contaminants from domestic, industrial and agricul-
tural wastewaters besides also they are impacted by overfishing,
harbor activities, and solid trash.
23,24
Santos estuary, in São
Paulo coast, is the most important Brazilian example of
environmental degradation from aquatic and atmospheric
pollution by industrial origin. The largest harbor in Latin
America (the Port of Santos) and the largest industrial complex
in Brazil are located in this area. Industrial activities began in
the 1950s with the establishment of diverse factories (steel, oil,
and agribusiness) and have turned this estuary into the final
destination for toxic waste and contaminated effluents since
then.
25
See Figure 1 for a map of the study areas.
Collected samples were taken from individual dolphins found
stranded dead at the beaches or incidental caught in fishing nets
along the Brazilian coast, Southwestern Atlantic, from 1994 to
2009. Available information on the samples is given in Table 1.
Sexual maturity is known to occur at different length depending
on the coastal area. The individuals considered in this study
included males and females, adult (sexually matured), juvenile
(sexually immature >100 cm length) and calves (sexually
immature <100 cm length) specimens.
Figure 1. Study area map, Southeaster and Southern coast of Brazil.
Brazilian States sampled: ES, Espi ́
rito Santo; RJ, Rio de Janeiro; SP,
São Paulo PR, Paraná; SC, Santa Catarina; RS, Rio Grande do Sul;
FMA, Franciscana Management Areas (I−IV).
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Table 1. Sampling Locations and Dates, Biological Information on the Dolphins Collected along the Brazilian Coast, And
Concentrations of OCT in the Liver Samples
a
location Brazilian State sample code sex length sexual maturity physical maturity sampling date (year) concentration OCT (ng g−1lw)
Espirito Santo (12) FT: 25% PON 08 M 70 Im Ca 2003−2006 nd
PON 12 M 73 Im Ca 2003−2006 129
PON 11 M 100 Im Ju 2003−2006 nd
PON 02 M 112 Im Ju 2003−2006 nd
PON 13 M 113 Im Ju 2003−2006 nd
PON 06 M 114 Im Ju 2003−2006 nd
PON 14 M 115 Ma Ad 2003−2006 nd
PON 09 M 117 Ma Ad 2003−2006 nd
PON 07 F 109 Im Ju 2003−2006 nd
PON 15 F 115 Im Ju 2003−2006 nd
PON 03 F 118 Im Ju 2003−2006 89
PON 10 F 136 Ma Ad 2003−2006 712
Rio de Janeiro (1) FT: 0% FT: 0% RJ 46 uk na na na 2003−2006 nd
Sao Paulo (10) FT: 70% BP 125 M 100 Im Ju 2006 100
BP 120 M 103 Im Ju 2005 nd
BP 133 M 112 Ma Ad 2006 380
BP 149 M 116 Ma Ad 2007 144
BP 176 M 122 Ma Ad 2008 141
BP 110 M 124 Ma Ad 2006 nd
BP 108 F 94 Im Ca 2006 524
BP 113 F 110 Im Ju 2006 269
BP 151 F 138 Ma Ad 2007 130
BP 140 F 110 Im Ju 2006 nd
Paraná(3) FT: 33% PR 50 F 56.5 Im Ca 2003−2006 129
PR 53 F 98 Im Ca 2003−2006 nd
PR 01 F 140 Ma Ad 2003−2006 nd
Santa Catarina (11) FT: 18% PB 221 M 83.5 Im Ca 2003−2006 nd
PB 23 M 102 Im Ju 2003−2006 nd
PB 22 M 107 Im Ju 2003−2006 nd
PB 53 M 87.3 Im Ca 2003−2006 345
PB 62 M102 Im Ju 2003−2006 401
PB 56 M109 Im Ju 2003−2006 nd
PB 222 F 129 Ma Ad 2003−2006 nd
PB 30 F 133 Ma Ad 2003−2006 nd
PB 37 F 133.5 Ma Ad 2003−2006 nd
PB 162 uk 127.5 Ma Ad 2003−2006 nd
PB 44 uk 145 Ma Ad 2003−2006 nd
Rio Grande do Sul (19) FT: 42% CA 143 M 125.5 na na 1997 nd
CA 32 M 129.5 Ma Ad 1994 nd
CA 142 M 133.7 Ma Ad 1997 nd
CA 36 M 137 Ma Ad 1994 153
CA 156 M 137 Ma Ad 1998 nd
CA 172 M 143 Ma Ad 1999 nd
CA 152 F 107.5 Im Ju 1998 142
CA 63 F 135.5 Im Ju 1994 nd
CA 124 F 137 Im Ju 1997 nd
CA 153 F na na na 1998 nd
CA 33 M 131 Ma Ad 1994 473
CA 108 F 157 Ma Ad 1995 nd
CA 173 F 161 Ma Ad 1999 493
CA 179 M 110 Im Ju 1999 107
CA 193 F 116 Im Ju 1999 129
CA 194 F 123 Im Ju 1999 782
CA 234 M 103 Im Ju 2000 103
CA 237 M 132 Im Ju 2000 nd
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Fifty six individual were analyzed, belonging to many States
of Brazil: Espi ́
rito Santo (n= 12), Rio de Janeiro (n= 1), São
Paulo (n= 10), Paraná(n= 3), Santa Catarina (n= 11) and
Rio Grande do Sul (n= 19). Liver samples collected were
placed in aluminum foil, frozen, and further lyophilized. Freeze-
dried liver tissue was ground, homogenized and stored in
brown glass sealed containers at −20 °C until analysis.
Analytical Methods. In order to prevent contamination
and photodegradation of samples and standard solutions all
glassware used was previously washed and heated overnight at
380 °C, and further sequentially rinsed with different organic
solvents and HPLC grade water. Separate solvents and only
previously unopened packages of solvents, chemicals and other
supplies were used. In addition, a set of at least two operational
blanks were processed together with each batch of samples.
Standard solutions and samples were always covered with
aluminum foil and stored in the dark. Furthermore, gloves were
worn during the sample preparation process.
Sample Preparation. Ssamples were extracted by PLE using
an automatic extractor ASE 200 (Dionex Corporation,
Sunnyvale, CA, USA). One gram dry weight of freeze-dried
dolphin liver tissue was mixed in the extraction cells with
hydromatrix. The PLE optimized parameters were as follows:
preheating of 5 min, heating of 5 min, two extraction cycles of
10 min using dichloromethane/hexane as extraction solvent (1/
1, v/v), temperature of 100 °C, pressure of 10 000 kPa, flush
volume of 80% of cell and 90 s of nitrogen purge. The PLE
extract obtained was concentrated to 3 mL and then subjected
to a purification step via acid attack with concentrated H2SO4
(95−97% purity) (4 ×2 mL). The extract was purified by SPE
with alumina cartridges (5 g/20 mL), using 40 mL of
hexane:dichloromethane (1/2). Finally, the extract was
evaporated to dryness. The residue was further reconstituted
with 0.1 mL of acetonitrile and the IS was added before LC-MS
analysis.
Lipid Content Determination. The lipid content determi-
nation was performed by gravimetric analysis. After the
extraction, the extracts were concentrated to incipient dryness,
each vial was weighed and the difference between the initial
weighing and weighing after the addition and evaporation rate
was used to calculate the percentage of lipids.
Percentage lipid content was determined for each individual.
Mean values were calculated for those specimens sampled in
the same geographical area, which were in the range 4% - 7%.
UPLC-ESI(+)-MS/MS Analysis. Target analysis of OCT was
performed by ultrahigh performance liquid chromatography
(UPLC)-tandem mass spectrometry (MS/MS) using an
Acquity UPLC chromatograph attached to a triple quadrupole
detector (TQD) mass spectrometer (Waters). A Hibar
Purospher STAR HR R-18 ec. (50 mm ×2.0 mm, 2 μm)
column (Merck) was used. The solvent flow rate was set to 0.4
mL min−1and the column temperature was kept at 50 °C. The
sample volume injected was 10 μL. The chromatographic
separation was performed by using as mobile phase HPLC
grade water (A) and acetonitrile (B), both with 0.3% formic
acid. The adopted elution gradient started with 5% of eluent B,
increasing to 95% in 1.20 min, kept constant for 2.30 min, and
rising to 100% in the following 0.5 min. During the next 2.5
min the elution gradient was kept constant, and then back to
initial conditions in 3 min.
MS/MS was operated in selected reaction monitoring
(SRM) and positive electrospray ionization mode (ESI+).
The optimized parameters were as follows: desolvation gas
(nitrogen), 750 L h−1; collision gas (argon), 0.19 mL min−1,
nebulization gas (nitrogen), 90 L h−1; ion spray voltage 3.35
kV; source temperature, 130 °C and desolvation temperature,
450 °C. Two characteristic transitions of the protonated
molecular ion [M+H]+(precursor ion) were recorded.
For the positive confirmation of OCT in liver tissue samples,
strict criteria had to be met in order to avoid false positives.
Following the European Commission Decision 2002/657/
EC,
26
that although it was initially conceived for food residue
analysis, it has been accepted by the scientific community for
environmental analysis, a minimum of three identification
points (IPs) is required for this purpose. In our case, these 3
IPs corresponded to the precursor ion (m/z362 amu) and to
the two transitions recorded from the precursor ion ([M+H]+)
to the product ions [M+H−C8H16]+and [M+H−C8H16−
H2O]+at m/z250 and 232 amu, respectively. Besides, the
chromatographic retention time of the analyte in the sample
should not vary more than 2.5% in comparison to the
calibration standards’, and the relative abundance of the two
SRM transitions monitored must also be compared to the
standards’corresponding values, and range about ±20%. Figure
2 represents the chromatograms for OC corresponding to a
standard solution at 40 ng mL−1, and to a dolphin liver sample
from an adult male from São Paulo. In this case, retention times
were exactly the same and the difference in SRM ratios was
solely 6%, and thus, confirming the identification
The described methodology probed to be precise and
sensitive for the quantification of OCT in dolphin liver samples
affording method limits of detection (LOD) and quantification
(LOQ) of 23 and 75 ng g−1lipid weight (lw), respectively, and
a relative standard deviation of 9%.
■RESULTS AND DISCUSSION
OCT Concentration in Liver Tissue. The analysis of the
samples revealed that OCT was present in 21 out of the 56
samples analyzed (38% frequency of detection) with concen-
trations in the range 89−782 ng g−1lw (see Table 1). These
concentrations are notoriously higher than that reported by
Balmer et al.
7
for OCT in lake fish (25 ng g−1lw). This
outcome was expected, as with other organic pollutants that
bioaccumulate and biomagnify along the food chain, given the
higher trophic level occupied by dolphins.
From the six sampling areas selected, Rio de Janeiro was one
of the areas where we expected to find residues of the sunscreen
because of its beach area with very active aquatic activities.
However, in the only sample taken OCT was not detected.
Despite that, we cannot rule out its presence in the area since
Table 1. continued
location Brazilian State sample code sex length sexual maturity physical maturity sampling date (year) concentration OCT (ng g−1lw)
CA 255 M 106 Im Ju 2001 nd
a
In parentheses, the number of samples analyzed, nd: not detected, uk: unknown, na: not available, FT: percentage of positive samples within the
area, Ca: Calves, Im: immature, Ma: mature, Ju: juvenile, Ad: adult. Method limit of detection (MLOD) and method limit of quantification (MLOQ)
are 23 and 75 ng g−1lw, respectively. Total frequency: 100 ×21/56 = 38%, Calves: 100 ×4/7 = 57%
Environmental Science & Technology Article
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we were only able to analyze one sample. In contrast, the most
contaminated zone was São Paulo, where OCT was most
frequently detected (70%), followed by Rio Grande do Sul,
where the UV filter was observed in 8 out of the 19 dolphins
sampled, and at the highest concentration, 782 ng g−1lw.
Nevertheless, the geographical distribution of positive samples,
as depicted in Figure 3, indicated that the highest mean
concentration (373 ng g−1lw) was determined in the samples
from Santa Catarina State. This sampling area is a partially
enclosed estuarine receiving industrial and urban wastewater
discharge, which could act as a sink for anthropogenic
pollutants. In a recent study with polybrominated diphenyl
ethers (PBDEs) in the same samples from this work, it was
observed also the higher levels in the dolphins from this
disturbed bay, in Santa Catarina.
20
Similar mean concentrations
were reported for OCT and for the group of PBDEs. The
comparison of the concentrations observed in the rest of areas
evidenced a different source for OCT and PBDEs anthro-
pogenic emissions. The geographical distribution of mean
concentrations (ng g−1lw) for OCT was: 373 (SC) > 310 (ES)
> 298 (RS) > 241 (SP) >129 (PR), whereas for the group of
congeners of PBDEs it was: 432 (SC) > 329 (SP) > 156 (PR)
> 144 (ES) >37 (RJ) > 34 (RS).
From 1994 to 2009 there has been a steady growing use of
UV filters as society has become aware of the dangerous effects
of sunlight. These currently popular chemicals have shown to
have a protective role against photoaging, photocarcinogenesis
and photoimmunosuppression promoted by UV sun radia-
tion.
27−29
Thus, potential temporal changes in the bioaccumu-
lation behavior of OCT were also assessed taking into account
the increasing use of sunscreen products. However, a direct
correlation could not be identified.
Relationships Between OCT Concentration and Bio-
logical Characteristics. The inclusion of individuals with
varying biological characteristics in this study provided an
opportunity to examine contaminant liver tissue concentrations
in relation to lipid content, sex, and physical and sexual
maturity.
The trend of increased concentration in biota samples with
increasing lipid content may be observed for a number of
organic pollutants.
30
In an attempt to assess the behavior of the
lipophilic UV filter, the correlation between liver lipid content
and OCT concentration was evaluated, however, no correlation
could be established based on these parameters (linear least-
squares regression coefficient of 0.371). The reason for this
differential behavior remains unclear.
Many studies have reported gender-specificdifferences in the
concentrations of persistent organic pollutants in marine
mammals, showing the well-known high variation in the
burden of lipophilic pollutants of females due to reproductive
stage.
20,31
Therefore, the potential correlation between sex and
OCT concentration was also assessed. Different statistical tests
were carried out, Pearson, Kendall, and Spearman. All they
showed no statistical differences among OCT concentrations
and gender. Significance values were: Pearson, 0.225; Kendall,
0.397; Spearman, 0.411.
Nevertheless, potential maternal transfer cannot be ruled out.
In order to assess the mother-to-calf transfer of OCT, a sample
of placenta from one pregnant female dolphin was collected
and analyzed, revealing that OCT was present in both the
placenta and the liver tissue at concentrations below LOQ (61
ng g−1Lw semiquantitative analysis) and 130 ng g−1lw,
respectively (liver sample reference BP151 in Table 1) being
indicative of gestational transfer. This hypothesis, however,
cannot be fully confirmed without data on a higher sample size
of pregnant female dolphins. However, it must be taken into
account the great difficulty in obtaining this kind of samples.
On the other hand, contamination data on breast milk could
also support the maternal transfer, specifically lactation transfer,
of the bioaccumulated OCT. This fact recently has already been
characterized in humans. Reported levels of OCT in women
breast milk were in the range 4.70−135 ng g−1lw, with 67%
frequency of detection.
32
This transfer was consistent with the
OCT accumulation data obtained in the present study, where
OCT was found in four out of the seven analyzed calves (57%
frequency of detection) higher than the frequency estimated
considering the complete set of individuals (38%), and at
increasing concentrations (129−524 ng g−1lw) as their length
was greater. Nevertheless, it has to be highlighted that,
Figure 2. Reconstructed SRM reference chromatogram for OCT
corresponding to a standard solution at 40 ng mL−1(a and b), and a
chromatogram corresponding to a dolphin liver sample from an adult
male (sample code BP 176) from Sao Paulo (c and d).
Figure 3. Distribution of OCT mean concentrations in dolphin’s liver
(ng g−1Lw) and standard error, along the Brazilian coast sampling
areas.
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obviously, the exposure pathways of human and dolphins are
clearly different.
Biomagnification. Several studies have probed that
biomagnification can occur for lipophilic organic contaminants.
In addition to high concentration levels, the process that
produce biomagnification also results in age (or length)-specific
patterns for adult male marine mammal. A different pattern has
been reported for adult female mammals despite being exposed
to contamination similarly to males. During gestation and
lactation contaminant body burden deceased by transfer to
calves.
20,33
In order to probe the potential biomagnification of OCT, a
full trophic analysis of this compound must be accomplished.
However, as lower trophic level organisms on these areas were
not available, we tried to provide preliminary evidence by
comparing our results with published data on the terrestrial
food chain. To date solely one study reported data on UV filter
biomagnifications. Fent et al.
3
recently investigated the
accumulation in the terrestrial food chain of EHMC, an UV
filter, having a similar lipophilicity (log Kow 6.1) to that of
OCT. The differences in EHMC concentrations in cormorant,
fish and macroinvertebrates suggested a trend for biomagnifi-
cation. Cormorants are migratory fish eating birds, representing
a high terrestrial trophic level. Average concentrations of
EHMC in five of these birds was 341 ng g−1lw, with values in
the range 16−701 ng g−1lw, whereas decreased average
concentrations were observed in lower trophic levels. This
concentration range was comparable to that obtained for OCT
in the present study (89−782 ng g−1lw). It must be pointed
out that cormorants are migratory birds with high metabolic
capacity and whose exposure routes in the terrestrial environ-
ment are different from that of the dolphins of the present
study, which are coastal and nonmigratory. Therefore, we
cannot perform an accurate comparison between these two
species.
In summary, these findings demonstrate for the first time that
the extensively used sunscreen agent OCT accumulates in liver
of dolphins at high concentration levels (up to 782 ng g−1lw)
similar to those of anthropogenic organic persistent pollutants.
This study also provides evidence that maternal transfer may
occurs trough placenta and likely also through breast milk.
The results presented herein suggest that OCT biomagnifies
through the marine food web. In order to probe the
biomagnification of this UV filter a full trophic level analysis
of OCT will be further performed. The present study
establishes the baseline levels for OCT in dolphins from
Brazilian coastal waters. Efforts should be directed toward the
analysis of other marine organisms to assess the impact of OCT
as well as other extensively used UV filters and their
transformation products on marine ecosystems.
■AUTHOR INFORMATION
Corresponding Author
*Phone: +34 93 400 6100; fax +34 93 204 59 04; e-mail:
sdcqam@cid.csic.es.
Notes
The authors declare no competing financial interest.
■ACKNOWLEDGMENTS
This research was funded by the Spanish Ministry of Economy
and Competitiveness through the Project CEMAGUA
(CGL2007-64551/HID). This work was also partly supported
by the Generalitat de Catalunya (Consolidated Research
Group: Water and Soil Quality Group 2009-SGR-965). Gago-
Ferrero acknowledges his fellowship to Junta para la
Ampliación de Estudios (JAE). This research was also funded
by the Ministry of Education of Brazil e CAPES (fellowship to
M.B. Alonso “Sandwich Programme”e PDEE; “Ciencias do
Mar”e Proc. 23038.051661/2009-18), Brazilian Research
Council e CNPq (Grant No. 304826/2008- 1), FAPERJ
(Jovem Cientista do Nosso Estado No. 101.449/2010), Mount
Sinai School of Medicine (NY/USA), Fogarty International
Center NIH/USA (grant 1D43TW0640). We are grateful to
the fishermen and cetacean research group staffs for the
assistance in fieldwork, as well as Cetacean Society Interna-
tional (CSI), Society for Marine Mammalogy (SMM) and Yaqu
Pacha. A.F.A. and J.L.-B. have research grant from CNPq (PQ-
2) and FAPERJ (JCNE). We give special thanks to students
from Environmental Chemistry Lab (IDAEA-CSIC, Spain),
Radioisotope Lab (UFRJ e Brazil) and Aquatic Mammal and
Bioindicator Lab (UERJ e Brazil).
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