Marine Pollution Bulletin 62 (2011) 15721576

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Marine Pollution Bulletin

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

Baseline

Levels and spatial distribution of polycyclic aromatic hydrocarbons (PAHs) in

sediments from Lenga Estuary, central Chile
Karla Pozo a,, Guido Perra a, Valentina Menchi a, Roberto Urrutia b, Oscar Parra b, Anny Rudolph c,
Silvano Focardi a
a
b
c

Environmental Science Department, University of Siena, Via Mattioli 4, 53100 Siena, Italy
Center for Environmental Sciences EULA-Chile, Universidad de Concepcin, Barrio Universitario s/n, casilla 160-C, Concepcin, Chile
Facultad de Ciencias, Universidad Catlica Santsima Concepcin, Alonso de Ribera 2850, P.C. 407 01 29 Concepcin, Chile

a r t i c l e
Keywords:
PAHs
Sediments
SQG
Lenga estuary
Chile

i n f o

a b s t r a c t
The Lenga Estuary is a small brackish wetland located southwest of San Vicente Bay, Region VIII, Chile.
Surface sediment from nine sites in the estuary were analysed for PAHs and compared to Sediment Quality Guidelines (SQG). Sediment samples were freeze dried and soxhlet extracted for 16 h using DCM.
Identication and quantication was carried out by HPLC. Organic carbon was also determined. Results
showed total PAH concentrations ranged from 290 to 6118 (2025 1975) ng g1 d.w. (2025 1975).
Results for organic carbon percentages ranged from 1% to 7%. Statistical analysis showed a signicant
positive correlation (Pearson test) between organic carbon percentage PAHs. Comparison of contaminant
levels and international Sediment Quality Guidelines (SQG) (ERL and ER) suggested that sediment of the
Lenga estuary did not show any ecotoxicologial risk for benthic organisms where high levels of PAHs
were detected. Monitoring of this and other contaminants is recommended in Chile.
2011 Elsevier Ltd. All rights reserved.

Estuaries are partly enclosed coastal bodies of water with one or

more rivers or streams owing into them with a free connection to
the open sea (Pritchard, 1967). Estuaries form a transition zone between river environments and ocean environments and are subject
to both marine inuences, such as tides, waves, and the inux of
saline water; and riverine inuences, such as ows of fresh water
and sediment. The inow of both seawater and freshwater provide
high levels of nutrients in both the water column and sediment,
making estuaries among the most productive natural habitats in
the world (McLusky and Elliot, 2004) being nurseries for coastal
sh and resting sites for migratory birds. In addition, because they
communicate with the sea, many estuaries have been used as
dumping sites for industrial and domestic efuents, turning them
into anaerobic digesters.
The Lenga estuary is a small coastal wetland (36470 0000 S;
73100 0000 W) in Region VIII, central Chile (Fig. 1). It is located in
the Hualpen Nature Sanctuary and is listed in the inventory of
the International Union for the Conservation of Nature (Scott and
Carbonel, 1986; Gysel and Lyon, 1987). Heavy industrialisation began in the 1950s, in the adjacent area of the estuary, in San Vicente
Bay, with construction of steelworks and an oil renery. The lack of
regulation in those days, combined with industrial growth, has had

Corresponding author. Tel.: +39 0577232879; fax: +30 0577232806.

E-mail address: gallardokarla@gmail.com (K. Pozo).
0025-326X/$ - see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.marpolbul.2011.04.037

a heavy impact on the coastal ecosystem. The estuary also acts as a

contaminant sink for the Biobio River, its major fresh water tributary and one of the most polluted water bodies in the country (Parra and Faranda, 1993). The river receives the efuent of mills that
produce 83% of Chilean wood pulp production (Gonzalez et al.
1999) and of sewage treatment plants as well as run-off from the
catchment area. Industrial activities and the river determine a considerable sediment load with high organic carbon content which
can act as an efcient pollutant trap.
Polycyclic aromatic hydrocarbons (PAHs) are persistent toxic
compounds that bioaccumulate in wildlife (UNEP, 2001). Contamination of PAHs in sediments may pose unacceptable risks for
aquatic organisms, wildlife and humans. Awareness of risks to humans and the environment from contaminated sediments can
inuence regulatory decisions (regarding, for example, dredging,
swimming, shing, shoreline rehabilitation). Because sediment
chemistry data alone does not provide an adequate basis for
assessing the hazards posed by sediment-associated contaminants
for aquatic organisms, interpretative tools are required to determine their potential concentration risk to the aquatic environment.
Sediment Quality Guidelines (SQG) provide a scientically justiable basis for evaluating the potential effects of sediment-associated contaminants on aquatic organisms. Sediment Quality
Guidelines (SQG), such as effects range low (ERL) and effects range
median (ERM), are valuable for establishing empirical relationships
between expected incidences of toxicity and the number and/or

K. Pozo et al. / Marine Pollution Bulletin 62 (2011) 15721576

1573

Fig. 1. Map of sampling sites in the Lenga estuary, Region VIII, central Chile.

degree to which SQGs values are exceeded. Recently, this approach

has been applied to others coastal sites around the world. For instance, Cardellicchio et al. (2007) used SQGs to provide possible
ecotoxicological risk estimations for benthic organisms at Mar Piccolo in Taranto, southern Italy and Binelli et al., 2008 reported a
comparison of SQGs for toxicity assessment in the Bay of Bengal
in India.
In Chile, various coastal ecosystems are subject to anthropogenic inuence but limited information is available on PAH levels.
Previous studies have documented contamination by PAHs in central Chile in lake sediments (Barra et al., 2004; Quiroz et al., 2005),
organisms (Barra et al., 2005) and marine environments (Mudge
and Seguel, 1999; Rudolph et al., 2002). In this study, we report
levels and distribution of PAHs in supercial sediments of the Lenga estuary. To evaluate the quality of sediments and to assess adverse biological effects in benthic organisms we used effectbased Sediment Quality Guidelines (SQGs) for PAHs.
Supercial sediments (1020 g) were obtained with a grab sampler in nine stations of the Lenga estuary during July 2002 (Fig. 1).
Sediment samples consisted mainly of coarse and medium sand
(76%) at most of the stations with the exceptions of stations 7, 8
and 9 where ne and muddy clays were dominant. Samples were
taken to the laboratory in an ice cooler, where they were weighed
and sealed to avoid contamination. Before analysis, sediments
were freeze dried at 50 C and 0.2 mbar. For extraction of PAHs,
dry sediments (510 g) were homogenised with clean anhydrous
sodium sulphate (previously extracted by a soxhlet system). Sediment samples were placed in pre-cleaned cellulose thimbles, and
extracted by soxhlet for 18 h with dichloromethane (DCM)
(300 ml). Extracts were percolated through activated copper to remove sulphur, concentrated by rotavapor and exchanged into hexane solvent (1 ml). For PAHs, clean-up extracts were concentrated
in a rotary evaporator to about 0.5 ml, and run on a silica gel column activated at 130 C over night. The silica column was slurry
packed with 20 ml of hexane. Then, PAHs extracts were eluted with
15 ml of DCM/Hexane (1:1) and reduced to 1 ml of acetronitrile
(Cousins and Jones, 1998 with some modications).
PAH identication and quantication was performed utilising
HPLC on a Waters (Milford, MA, USA) liquid chromatographic

system, with a Waters 474 Scanning Fluorescence Detector and

Waters 996 Photodiode Array Detector, using a Supelcosil-PAHs
column (250  4.6-mm I.D., particle size 5 lm). A mobile phase
ow rate of 1.5 ml/min was used with the following linear gradient
elution program: acetonitrile:water (v:v) in a linear gradient from
40:60 in 1 min to 100:0 in 25 min maintaining this ratio for 15 min
and going back to 40:60 in 4 min. The column temperature was
maintained at 25 C. Each sample was quantied for 16 PAHs proposed by EPA as priority pollutants (see Table 1). Fluorescence
detection was optimised with wavelength (kex(nm)) programming
for the excitation and emission as follow: Naph (220330), Ace
(270323), Fl, Phen, Ant (248374), Flu (237460), Pyr, BaA, Chr
(270400), BbF, BkF, BaP, DBA, BgP (290418) and IP (270490).
Acenaphthylene has the weakest uorescence and must be determined by UV absorbance. PAHs concentrations were expressed in
ng g1 d.w.
The above procedures were checked for recoveries and reproducibility. Procedural blanks and reference material purchased
from the National Institute of Standards and Technology (NIST)
was analysed for QA/QC purposes. Analysis of PAH reference material (HS-6) showed a mean recovery of 95%. Prior to sediment
extraction, six analytical blanks were prepared using the same
extraction and clean-up procedure. A solvent blank was analysed
after every 15 samples to check the chromatographic response.
Limits of detection (LODs) were dened as the average blank
(n = 4) plus three standard deviations (SD). When target compounds were not detected in blanks, 2/3 of the instrumental detection limit was used as the method detection limit (MDL). All
qualied data (i.e., exceeding the MDL) were blank corrected. MDLs
ranged from 0.01 to 0.5 ng g1 for PAHs.
In order to evaluate the possibility of adverse effects in sediments of the Lenga estuary, concentrations of PAHs were compared
to international SQGs. SQGs were limited to the use of the effects
range low (ERL) and effects range median (ERM) weight-of-evidence approach of the National Oceanic and Atmospheric Administration (NOAA) (Long et al., 1995; MacDonald et al., 1996).
Organic carbon analysis was carried out with 0.10.5 g of dry
sediment thoroughly mixed with 10 ml potassium dichromate
(K2CrO7, 1 N). Concentrated sulphuric acid (20 ml) was added,

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K. Pozo et al. / Marine Pollution Bulletin 62 (2011) 15721576

Table 1
Concentrations (ng g1 d.w.) of 16 priority polycyclic aromatic hydrocarbons (PAHs) and organic carbon (%) in supercial sediments of Lenga estuary.
Compound/station

MDL

L1

L2

L3

L4

L5

L6

L7

L8

L9

Naphthalene
Acenaphthylene
Acenaphtene
Fluorene
Phenanthrene
Anthracene
Fluoranthrene
Pyrene
Benz[a]anthracene
Chrysene
Benzo[b]uoranthene
Benzo[k]uoranthene
Benzo[a]pyrene
Indeno[1,2,3-cd]pyrene
Dibenzo[a,h]anthracene
Benzo[g,h,i]pyrene

0.25
0.50
0.05
0.04
0.01
0.01
0.01
0.04
0.02
0.02
0.01
0.01
0.02
0.02
0.10
0.03

6
bdl
106
10
131
45
390
369
123
173
164
85
207
87
108
128

bdl
bdl
6
3
36
11
86
109
41
51
51
25
56
4
23
50

3
bdl
18
3
41
10
120
91
58
82
72
32
111
9
59
47

5
bdl
347
bdl
334
120
798
1017
247
124
254
209
313
167
161
228

15
bdl
60
13
116
40
350
272
166
224
200
76
226
110
135
130

15
bdl
bdl
3
18
6
56
36
22
21
24
13
22
15
18
21

29
bdl
134
40
440
180
1050
953
425
457
575
234
564
327
312
398

7
bdl
41
5
49
19
140
86
47
54
128
25
68
17
27
45

16
bdl
73
10
114
38
215
144
86
108
82
36
121
10
48
62

2131
133
113
1

551
39
31
2

757
50
39
2

4323
309
273
7

2134
142
98
4

290
21
13
2

6118
408
295
4

757
50
40
1

1162
77
57
1

Tot PAHsa
Mean
SD
Organic carbon (%)

Abbreviations: MDL, method detection limit; bdl, below detection limit; SD; standard deviation.
a
RPAH16: 2-ring PAHs include naphthalene; 3-ring PAHs include acenaphthylene, acenaphthene, uorene, phenanthrene and anthracene; 4-ring PAHs include uoranthene, pyrene, benzo[a]anthracene and chrysene; 5-ring PAHs include benzo[b]uoranthene, benzo[k]uoranthene, benzo[a]pyrene and dibenzo[ah]anthracene; 6-ring
PAHs include indeno[1,2,3-cd]pyrene and benzo[ghi]pyrene.

Concentrations (ng/g d.w)

distribution patterns shows that the highest PAH concentrations

(ng g1 d.w.) were found at stations L7 (6118), L4 (4319) and L5
(2134) (Fig. 2a). These results are higher than PAH levels detected
in other coastal areas of Chile. For instance, Mudge and Seguel
(1999) reported 1210 ng g1 d.w. in sediments of San Vicente
Bay. Rudolph et al. (2002) reported PAH concentration (lg/kg) of
anthracene (<0.05159), phenanthrene (64281), uoranthrene
(141706) and pyrene (<0.05572) in sediments of Concepcin
Bay, central Chile. However, PAH levels detected in this study are
similar to those of highly PAH-polluted sites in other parts of the
world. Tan and Heit (1981) reported levels up to 12,100 ng g1 in
Lake Woods, USA; Hong et al. (1995) found values of 1200
14,000 ng g1 in Victoria Harbour, Hong Kong, China; Baumard
et al. (1998) detected levels of 9004100 ng g1 in Arcachon Bay,
France; Meniconi et al. (2002) found 918035 ng g1 in Guanabara

Total PAHs

7000
6000
5000
4000
3000
2000
1000
0

L1

L2

L3

L4

L5

L6

L7

L8

L9

Organic Carbon (%)

Percentage (%)

7
6
5

60

4
3
2
1
L4

Fig. 2. PAH concentrations (ng g

of the Lenga estuary.

L5

L6

L7

L8

L9

40

d.w.) (a) and OC (%) (b) in supercial sediments

and the mixture was shaken for 30 s and left to rest for 30 min. The
sample was diluted to 100 ml with distilled water containing 10 ml
phosphoric acid (H3PO4) and 0.2 g sodium uoride (NaF). Finally,
samples were cooled and titrated with ferrous ammonium sulphate Fe8(NH4)2(SO4)2 0.5 N (Gaudette et al., 1974; Pozo et al.,
2007, 2009). The statistical analysis was performed with the SAS
statistical software package (SAS, Version 7). Pearson correlation
was used to analyse relationships between PAHs and organic carbon percentage (%).
Table 1 shows PAH concentrations and carbon content (%) in
sediments of the Lenga estuary. Total concentrations of PAHs
(ng g1 d.w.) ranged from 290 to 6118 (2025 1975). Spatial

20

0
6RPAHs

1

5RPAHs

L3

4RPAHs

L2

2RPAHs

L1

3RPAHs

L9

L8

L7

L6

L5

L4

L3 L2

L1

Fig. 3. Homologue composition (%) of polycyclic aromatic hydrocarbons (PAHs) in

sediment from the Lenga estuary.

K. Pozo et al. / Marine Pollution Bulletin 62 (2011) 15721576

Bay in Brazil; and Medeiros et al. (2005) detected a range of 38

11,780 ng g1 at Patos Lagoon estuary in Brazil.
PAH composition pattern was dominated by four- (48%) and
ve-ring (26%) PAHs (Fig. 3). Of the individual PAHs analysed, uoranthene and pyrene accounted for 40% and 30% of total PAHs
composition, respectively. Ratios of phenanthrene/anthracene
and uoranthene/pyrene were also estimated in order to distinguish between PAHs of different origin. The phenanthrene/anthracene ratio is temperature-dependent and is approximately 3 for
emissions from the combustion of various fuels (Gschwend and
Hites, 1981). Predominance of uoranthene/pyrene is classically
of pyrolytic origin, namely coal combustion (Simo et al., 1997). In
the Lenga estuary, the average PAH ratio was 3 for phenanthrene/anthracene and 1.2 for uoranthene/pyrene, indicating a
pyrolytic origin of the predominant PAH compounds.
Interesting results were also obtained for potentially carcinogenic PAHs (benzo[a]anthracene + benzo[b]uoranthene + benzo[k]uoranthene + benzo[a]pyrene + indeno[1.2.3-cd]pyrene + dibenzo
[ah]anthracene) with concentrations (ng g1 d.w.) ranging from 114 to
2437 accounting for 3354% of the total PAHs composition. In particular, pyrene and benzo[a]pyrene showed high levels at stations L4
and L7 (Table 1). It was also noteworthy that naphthalene, the simplest low molecular weight PAH, was found at relatively high levels,
suggesting that its contribution was mainly from anthropogenic
sources.
Evaluation of potential ecotoxicological risk was also estimated.
SQG calculations such as effects range low (ERL) and effects range
median (ERM) were used to evaluate sediment quality in relation
to PAH levels in the Lenga estuary. Table 2 presents the concentration ranges of PAHs proposed by international SQGs (ERLERM)
and the number of stations amongst ranges of international SQGs
at Lenga estuary. SQG values showed that PAHs exceeded the
ERL for total PAHs (4.0  103 ng g1 d.w.) only at two stations
(L4 = 4323 and L7 = 6118), and no stations exceeded the ERM
(44  103 ng g1 d.w.). With the exception of dibenzo[ah]anthracene at station L1, individual PAHs did not exceed the ERL
(Table 2).
Organic carbon percentage (OC%) uctuated between 1% (L1)
and 7% (L4) (Table 1 and Fig. 2b). These results are in agreement
with the OC% reported in other coastal areas of Chile. Moscoso
et al. (2006) reported OC% of 1.96.8% in the Lenga estuary, Silva
et al. (1998) detected average OC% of 5.8 1.8 in Chilean Fjiords,
Southern Chile and Aguirre-Martnez et al. (2009) found OC% about
6% in Caleta ColiumoCentral Chile. Statistical analyses showed
signicant positive correlations between OC% and PAH
Table 2
SQGs values for PAHs and number of stations amongst ranges of international
Sediment Quality Guidelines at Lenga estuary.
Compound
Naph
Acy
Ace
Fl
Phen
Anthr
Fluo
Pyr
B[a]A
Chry
B[b + k]Fl
B[a]P
IDP
DB[ah]A
B[ghi]P
RPAHs
a

SQGa ERLERM (ng g1 d.w.)

1602100
44640
16500
19540
2401500
851100
6005100
6652600
2611600
3842800

4301600

63260

402244,792

No. of stations
<ERL
ERLERM
L9
L9
L2
L8
L7
L7
L7
L7
L8
L8

L8
L5

L7

L7
L1
L2
L2
L2
L2
L1
L1

L1

L3

L2

>ERM

L1

SQG values taken from Long et al. (1995) and MacDonald et al. (1996).

1575

concentrations (r = 0.73; p < 0.05) at the Lenga estuary. These results indicated that OC content is a good indicator of adsorption
of organic contaminants by suspended particulate matter. Consequently, the subsequent sedimentation process is also an important factor that enhances the transport, diffusion and fate of
these compounds in the estuary.
In conclusion, these results show that concentrations of PAHs
were high and similar to those recorded in others highly PAH polluted sites of the world. The ecotoxicological evaluation based on
international SQGs indicates no ecotoxicological risk for benthic
organisms exposed to relatively high levels of PAHs. However,
damage might also occur at lower concentrations than those used
as guidelines. For instance, Thomas et al. (1999) reported potential
physiological tolerance for those organisms exposed chronically to
PAHs levels. Further studies for other potential contaminants (i.e.,
pesticides, PCBs, PCDD/Fs) are recommended in other coastal ecosystems in Chile. In addition, education of the general public about
the hazards associated with production of toxic compounds is
essential for long-term mitigation of potential environmental
problems. These results are an important contribution to knowledge of levels of PAHs in estuary ecosystems in Chile.
Acknowledgements
This work was supported by the international collaboration
agreement between Siena University and Concepcin University,
which permits the academic interchanges and to conduct research
in the Biobo and Patagonia Regions in Chile. We also want to thank
UNESCO-EOLSS Chair in Natural Resource Management, Land
Planning and Environmental Protection and to project Fondecyt
N1070508 and N1080294 for their partial support.
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