Aquatic Toxicology 98 (2010) 83–90

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Aquatic Toxicology
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Bioavailability of sorbed phenanthrene and permethrin in sediments to

Chironomus tentans
Xinyi Cui a,b , Wesley Hunter b , Yu Yang b , Yingxu Chen a , Jay Gan b,∗
a
Institute of Environmental Science and Technology, Zhejiang University, Hangzhou 310029, PR China
b
Department of Environmental Sciences, University of California, Riverside, CA 92521, USA

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

Article history: The availability of sorbed hydrophobic organic contaminants (HOCs) to benthic organisms is important
Received 17 November 2009 for characterizing sediment toxicity. While many studies show a correlation between the rapid desorp-
Received in revised form 14 January 2010 tion HOC pool and bioavailability to benthic organisms, bioavailability of the slow or very slow desorption
Accepted 19 January 2010
fraction is still poorly understood. In this study, Chironomus tentans were exposed to phenanthrene (PHE)
or permethrin (PM) to derive biota-sediment accumulation factors (BSAFs) in a sediment that was sequen-
Keywords:
tially desorbed with Tenax extraction or amended with a charcoal to modify the distribution of PHE and
Bioavailability
PM among the rapid (frapid ), slow (fslow ) and very slow (fvslow ) desorption pools. As the desorption interval
Desorption
Chironomus tentans
was increased, the frapid quickly decreased to zero and became negligible after 12 h desorption for PHE and
Phenanthrene 48 h desorption for PM. However, in samples with depleted frapid , BSAF values were substantially greater
Permethrin than zero, suggesting availability of fslow and fvslow . A multivariate linear regression model was further
Sediment toxicity used to estimate BSAFs specific to the different desorption pools, i.e., BSAFrapid , BSAFslow or BSAFvslow . The
slow desorption pool was found to be readily available to C. tentans, with BSAFslow values ranging from
25.3 to 73.9% of BSAFrapid . In comparison, BSAFvslow ranged from 0 to 5.9% of BSAFrapid , suggesting a lack of
availability. Therefore, the kinetically slow desorption fraction is relatively bioavailable and should not
be ignored in sediment toxicity assessment.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction toxicity to small aquatic invertebrates (Kukkonen et al., 2004;

The bed sediment serves as the depository of hydrophobic
organic contaminants (HOCs) in an aquatic environment (Muir et
al., 1994; Kukkonen and Landrum, 1998). Therefore, the threat
posed by contaminated sediments is an important component of
risk assessment for HOCs in aquatic ecosystems. However, risk
assessment of sediment-associated contaminants has been proven
a difficult task because many factors, such as the heterogeneous
nature of sediment organic carbon (Di Toro et al., 1991; Luthy et al.,
1997; Knauer et al., 2007) and animal behavior (Gobas et al., 1993),
affect the bioavailability of sediment-sorbed HOCs.
The role of desorption in bioavailability has been recognized in
many studies (Kraaij et al., 2001; Leppänen and Kukkonen, 2006).
Desorption of HOCs from the sediment phase is generally char-
acterized by multiple stages, reflecting pools of chemicals sorbed
at different strengths (Cornelissen et al., 1998). It is generally
believed that only the rapid desorption fraction (frapid ) is bioavail-
able in diffusion-driven processes such as biodegradation and acute
∗ Corresponding author at: University of California, 900 University Avenue,
Riverside, CA 92521, USA. Tel.: +1 951 827 2712; fax: +1 951 827 2712.
E-mail address: jgan@ucr.edu (J. Gan).
Reichenberg and Mayer, 2006; ten Hulscher et al., 2003). While
the availability of frapid continues to be shown in an increasingly
number of studies, relatively little is known about the bioavailabil-
ity of slow (fslow ) or very slow (fvslow ) desorption fraction to benthic
organisms. Several studies suggested that accumulation of HOCs by
benthic organisms could occur either from liquid phase or dietary
exposure (Comber et al., 2008; Mcleod et al., 2008). Some in vitro
experiments further showed that particle-associated HOCs could
be desorbed by digestive fluid during passage in the gut (Mayer et
al., 2001). Therefore, the role of fslow and fvslow in bioaccumulation
may be influenced by biological factors, such as sediment forage
activity, assimilation efficiency, and gut fluid extraction. Such inter-
actions also suggest that contributions from fslow and fvslow may
be reduced, but should not be negligible. For instance, Kraaij et al.
(2002) demonstrated that the concentration of rapidly desorbing
phenanthrene in the sediment after 48-h Tenax extraction declined
by an average factor of 27, while the BSAF value was lower only by
a factor of 2. However, in general, there is still a lack of experimen-
tal evidence and also a quantitative approach to ascertaining the
availability of HOCs in fslow and fvslow .
The objective of this study was to quantitatively evaluate the
availability of HOCs in fslow and fvslow to Chironomus tentans. The
biota-sediment accumulation factor (BSAF) of a PAH compound

0166-445X/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.aquatox.2010.01.016
84 X. Cui et al. / Aquatic Toxicology 98 (2010) 83–90
(phenanthrene) and a pyrethroid insecticide (permethrin) was
measured in a sediment following sequential extraction with Tenax
beads or amendment with a charcoal to modify the relative sizes
of frapid , fslow , and fvslow . A model was further developed and used
to derive BSAFs specific to the individual desorption pools, i.e.,
BSAFrapid , BSAFslow and BSAFvslow , to discern the availability of fslow
and fvslow . The estimation of BSAF specific to desorption pools is a
novel approach and may be generally used for evaluating bioavail-
ability of sediment or soil-sorbed contaminants.
2. Materials and methods
2.1. Chemicals and materials
14 C-Carbonyl-labeled permethrin (PM) (0.3–0.5 mCi/mmol) was
provided by FMC (Philadelphia, PA, USA) and purified using thin-
layer chromatography before use. The stock solution was made in
acetone with concentration of 171 mg/l. 14 C-9-Phenanthrene (PHE)
(≥95% radiochemical purity, 40–60 mCi/mmol) was obtained from
Sigma–Aldrich (St. Louis, MO, USA). The stock solution was also
made in acetone with concentration of 100 mg/l. Tenax TA (60–80
mesh) was purchased from Supelco (Bellefonte, PA). A charcoal
derived from coconut shell (Agrocoir, Laguna Niguel, CA, USA) was
used as a representative black carbon (BC) material to amend the
sediment. All solvents and other chemicals used in the study were
gas chromatography (GC) or analytical grade.
2.2. Sediment treatment and equilibration
The sediment was sampled from the top 10 cm in San Diego
Creek, located in Orange County, CA, USA. The collected sediment
was wet sieved through a 2 mm mesh, and stored at 4 ◦ C before use.
The total organic carbon (TOC) content was 0.4%. The background
BC content was 0.03% based on sediment dry weight, which was
much lower than the charcoal amendment rate (0.4%) used in this
study. TOC was determined by removing inorganic carbon using
digestion with HCl (1 M) and analysis of the digested sample by
combustion on an elemental analyzer (Flash EA1112 Combustion
Nitrogen/Carbon Analyzer System, Thermo Finnigan, Woods Hole,
MA, USA). The BC content was determined following the method of
Gustafsson et al. (1997). Briefly, dried and finely ground (<250 ␮m
particle size) sediment was combusted in a muffle furnace at 375 ◦ C
for 24 h. Samples were then digested with HCl until effervescence
ceased, and the BC content was analyzed on the elemental analyzer.
Sediment, with or without charcoal amendment, was spiked
with 14 C-PM or 14 C-PHE at 100 ␮g/kg (dry weight) and equilibrated
for 21 d before subjecting to desorption. The spiking procedure
consisted of placing a small amount of silica sand inside a 1.9 l
wide-mouth glass jar and treating with 14 C compounds. The sedi-
ments were spiked by addition of acetone solutions containing the
test compounds (final ratios were 0.1 ml of acetone/100 g of dry
sediment in phenanthrene-spiked treatments and 0.06 ml of ace-
tone/100 g of dry sediment in permethrin-spiked treatments). After
the carrier solvent was allowed to evaporate for about 1 h in a fume
hood until dry, 500 g (dry weight) of sediment was mixed thor-
oughly with the spiked sand. In a separate set of tests, the sediment
was amended with 2 g finely ground (<250 ␮m particle size) char-
coal (0.4%, w/w) for the purpose of evaluating the effect of BC on
the desorption and bioavailability of PM and PHE. Deionized water
was used to adjust the water-to-sediment ratio to about 1:1 (w/w).
The treated sediment samples were capped and shaken at 80 rpm
in the dark at room temperature for 21 d.
To check if the spiked chemicals were evenly distributed,
aliquots of 0.1 g (dry weight) treated sediments were removed and
air dried in a fume hood for 24 h. The subsamples were combusted
at 900 ◦ C on an OX-500 biological oxidizer (R.J. Harvey, Hillsdale,
NJ, USA) for 4 min. The evolved 14 CO2 was trapped in 15 ml of
Carbon-14 cocktail (R.J. Harvey) and the radioactivity was mea-
sured on a Beckman LS 5000TD liquid scintillation counter (LSC)
(Fullerton, CA, USA). The relative standard deviation of radioactiv-
ity per gram dry sediment from replicates was 0.9–3.6%, suggesting
uniform distribution of PHE and PM in the spiked sediments.
2.3. Tenax-aided desorption
The equilibrated sediments were desorbed for different time
increments using a method similar to that described in Xu et al.
(2008) to modify the relative sizes of frapid , fslow and fvslow . Briefly,
11 g (dry weight) aliquots of sediment were transferred to 250 ml
polyethylene centrifuge tubes. Tenax beads (0.30 g) and 100 ml
deionized water were then added to each container. The samples
were shaken on a horizontal shaker at about 100 rpm at room
temperature for 1, 2, 6, 12, 24, 48, 96, 144, 192, 240 or 312 h,
with four replicates used for each desorption interval. The sam-
ple slurries were centrifuged at 3000 rpm for 20 min, followed by
filtration of the supernatant using a Whatman No. 41 filter paper
(Whatman, Maidstone, UK) to recover the Tenax beads. The trapped
Tenax beads were rinsed thoroughly with deionized water and
were extracted by sonication using 3 ml of acetone–hexane mix-
ture (1:1, v/v) in 20 ml glass scintillation vials for three consecutive
times. The sonication was conducted for 5 min in 2 s pulse mode
by a high intensity ultrasonic processor (Sonic 550, Fisher). The
extracts from the same sample were combined and mixed with
10 ml Ultima Gold LSC cocktail (Fisher), after which the 14 C radioac-
tivity was measured by LSC to derive the desorbed amount of PHE
or PM.
The desorption kinetics up to 312 h were used to construct the
desorption curve and for estimating the initial rapid, slow, and very
slow desorption fractions (Frapid , Fslow , and Fvslow ) by fitting data to
a three-phase model (Cornelissen et al., 1997), using SigmaPlot 10.0
(San Jose, CA, USA):
St
= Frapid e−krapid t + Fslow e−kslow t + Fvslow e−kvslow t (1)
S0
where St and S0 (ng/g) are concentrations of compounds in the sed-
iment after desorption time interval t (h) and before desorption,
respectively, and krapid , kslow , and kvslow are the rate constants for
the rapid, slow and very slow desorption fractions, respectively.
2.4. Bioaccumulation experiment
The freshwater invertebrate C. tentans (Aquatic Biosystems, Fort
Collins, CO, USA) was used in the bioaccumulation experiment. The
C. tentans were cultivated in the laboratory using reconstituted hard
water (RHW) and silica sand as the substrate for several months
at 23 ± 2 ◦ C before use. The RHW contained NaHCO3 (192 mg/l),
CaSO4 ·2H2 O (120 mg/l), MgSO4 (120 mg/l), and KCl (8 mg/l). The
photoperiod for organism stock cultures was 16:8 h light:dark. The
C. tentans were fed once a week with 80 ml slurry of TetraFinTM
Goldfish Flakes (Koi Lagoon, Fort Collins, CO, USA) with concentra-
tion of 100 mg/l.
The biota-sediment bioaccumulation factor (BSAF) was mea-
sured using C. tentans and the above incrementally desorbed
sediments. Briefly, the four replicates of Tenax-extracted sediment
after each time interval were transferred to a 75 ml aluminum foil
container (Fisher) followed by addition of 30 ml of RHW. After the
sediment settled overnight, 10 C. tentans (fourth-instar, based on
head capsule width) were introduced into each test vessel. Organ-
isms were harvested after 24 h of exposure by filtering the sediment
through a 1-mm sieve. Previous studies showed that the uptake of
PHE and PM by C. tentans reached equilibrium within 24 h (Hunter
 X. Cui et al. / Aquatic Toxicology 98 (2010) 83–90 85

et al., 2008; Supporting Information Fig. S1). The C. tentans were

rinsed in deionized water and placed on a piece of pre-weighed
paper tissue to air dry for 24 h. The dried organisms and paper
were weighed again to obtain the dry weight of organisms. About
0.05 g (dry weight) of the sediment was also transferred onto an alu-
minum weighing dish (Fisher) and air-dried at room temperature
in a fume hood for 24 h. Dry C. tentans and sediment samples were
combusted at 900 ◦ C for 2 and 4 min, respectively, on the biological
oxidizer, and the trapped 14 CO2 was analyzed by LSC to derive the
chemical mass in C. tentans (Mb ).
The chemical associated with the sediment in the gut content
(Mg ) was excluded in calculating BSAF. Mg was determined using
the method recommended by the U.S. EPA (USEPA, 2000). Briefly,
C. tentans after 24 h exposure in the sediment–water systems were
rinsed in deionized water and placed on a pre-weighed weighing
dish to air dry. Before use, the weighing dishes were burned at
550 ◦ C for 2 h in a muffle oven to eliminate weighing errors due to
the pan oxidization during combustion. Dry organisms and weigh-
ing dishes were weighed again to obtain the dry weight of the
organisms. The dried organisms in the dish were then ashed at
550 ◦ C for 2 h in a muffle oven. The dish with the ashed residue
was then weighed again and the sediment mass in gut content was
determined as the mass of residue after burning. The average gut
content for C. tentans determined in this manner was 11.8% of the
dry body weight, which was similar to the 10% value reported in
Brooke et al. (1996). The average sediment content in the gut was
used in the calculation of BSAF:
(M b − Mg )/mbio
BSAF = (2)
Cs
Mg = 11.8% × mbio × Cs (3)

where mbio is the dry weight of C. tentans, and Cs is the chemical

concentration in the sediment (g/g dry weight).

2.5. Modeling contributions from various fractions

Under the assumption that all three phases independently con-
tributed to the total bioaccumulation, the overall BSAF may be
described by the following relationships:
Fig. 1. Desorption kinetics of (a) phenanthrene and (b) permethrin from sediment
(䊉) or charcoal-amended sediment (). St /S0 is the ratio of chemical mass remain-
ing in the sediment after desorption time t to that present in the sediment before
desorption.

BSAF = BSAFrapid · frapid + BSAFslow · fslow + BSAFvslow · fvslow (4)

S0
frapid = Frapid e−krapid t ·
St
S0
fslow = Fslow e−kslow t ·
St
S0
fvslow = Fvslow e−kvslow t ·
St
where BSAFrapid , BSAFslow and BSAFvslow are the biota-sediment
accumulation factors for the rapid, slow, and very slow desorp-
tion fractions, respectively, frapid , fslow , and fvslow are rapid, slow,
and very slow desorption fractions remaining, respectively, after
desorption time t (h). The values of frapid , fslow , and fvslow could
be calculated based on the parameters obtained from Tenax-
desorption test (Eq. (1)). The frapid , fslow , and fvslow together with
BSAF at each desorption time interval were then subject to statisti-
cal analysis of linear regression (SPSS 10.0) to obtain the regression
coefficients BSAFrapid , BSAFslow and BSAFvslow.
2.6. Quality assurance and control
Several quality control measures were used in this study. First,
selected sediments were freshly spiked with 1000 ␮g/kg 14 C-PM
and tested for 14 C recovery for the sample combustion method. The
recovery was found to be 80.1 ± 7.9%, and the averaged recovery
was used for correction. Moreover, PHE and PM recoveries for the
(5) extraction of Tenax beads were determined. Briefly, 25 ml of Milli-Q
water containing known chemical concentrations (100 ␮g/l) were
extracted with Tenax in triplicate following the same procedure as
(6)
described above with 2 h of shaking time. The Tenax beads were
separated from the water, extracted with acetone–hexane mix-
(7) ture (1:1, v/v), and analyzed by LSC. Recoveries of 89.1–98.3% were
obtained for PHE, and 90.2–97.5% for PM.
3. Results and discussion
3.1. Desorption kinetics
The spiked sediments, with or without charcoal amendment,
were desorbed for different time intervals, and the plots of St /S0 as
a function of desorption time interval t are shown in Fig. 1. The over-
all desorption kinetics consisted of an initial rapidly declining stage,
followed by a transitional phase and then by a very slowly declining
stage. The triphasic desorption model accurately described the des-
orption kinetics for all treatments, with r2 ≥0.99 (p < 0.05) (Table 1).
In the non-amended sediment, desorption of PHE was much faster
than that of PM. For instance, after 312 h desorption, 67.4% of PHE
was desorbed, as compared to 27.5% for PM. As the log Kow val-
ues of PHE and PM are 4.45 (Karickhoff, 1981) and 6.10 (Laskowski,
2002), respectively, the difference may be attributed to the fact that
86 X. Cui et al. / Aquatic Toxicology 98 (2010) 83–90

Table 1
Mean regression parameters from fitting Tenax desorption data of phenanthrene and permethrin in non-amended and charcoal-amended sediments to a triphasic desorption
model.

Treatment Frapid a krapid (h−1 ) Fslow kslow (h−1 ) Fvslow kvslow (h−1 )

Phenanthrene
Non-amended 0.265 3.80 × 100 0.339 4.18 × 10−2 0.397 6.54 × 10−4
Charcoal-amended 0.131 4.95 × 10−1 0.151 2.99 × 10−2 0.719 2.65 × 10−13

Permethrin
Non-amended 0.114 9.9 × 10−2 0.172 7.32 × 10−3 0.704 8.92 × 10−14
Charcoal-amended 0.124 7.7 × 10−2 0.422 3.50 × 10−3 0.434 6.49 × 10−12
a
Frapid , Fslow , and Fvslow are the rapid, slow, and very slow desorption fractions, respectively, and krapid , kslow , and kvslow are the corresponding rate constants.

more hydrophobic chemicals tend to have lower molecular diffu-
sion rates (Cornelissen et al., 1997; You et al., 2007). Cornelissen et
al. (1997) further suggested that desorption of HOCs depended on
the size of molecules. Given that PHE has a molar molecular volume
of 115.4 cm3 while PM has a much higher value at 237.4 cm3 (Yang
et al., 2009), our results further validated the potential importance
of molecular size in the desorption of HOCs from sorbents such as
sediments.
Amendment of charcoal resulted in a pronounced reduction in
PHE desorption from the sediment, but a similar effect was not
observed for PM (Fig. 1). Unlike PHE, desorption of PM after char-
coal addition increased slightly compared to the non-amended
sediment, although paired t-test showed the difference to be
insignificant. The derived Frapid in the charcoal-amended sediment
was only a half of that in the non-amended sediment for PHE, but
the difference was not significant for PM (Table 1). Concurrently,
after charcoal amendment, the fraction of PHE in the slow desorp-
tion pool decreased (Fslow ) while that in the very slow desorption
pool (Fvslow ) increased significantly (p < 0.01) from 0.397 to 0.719.
In comparison, in the charcoal-amended sediment, partition of PM
shifted toward the slow desorption pool while Fvslow decreased sig-
nificantly from 0.704 to 0.434. The lack of a significant effect of
charcoal addition on PM desorption was inconsistent with previ-
ous findings for PAHs, PCBs and hexachlorobenzene (Chai et al.,
2008; Oen et al., 2006; Sun and Ghosh, 2008), but was in agree-
ment with the findings on other chemicals. For instance, adsorption
of pyrethroid compounds including PM on pure charcoal was found
to be similar to that on sediment organic carbon in Yang et al.
(2009), while no significant effect by BC was observed for atrazine
in Cornelissen et al. (2005). The cause may be again attributed to
the relatively large molecular size of PM, which could have inhib-
ited the diffusion of PM molecules into the nanopores of charcoal
(Yang et al., 2009). Sun et al. (2009) also reported that larger PCB
congeners (e.g., PCB 153) were transferred more slowly from the
sediment phase to activated carbon.
The makeup of the remaining sorbed chemical as a function of
desorption time was calculated based on the parameters listed in
Table 1, using Eqs. (5)–(7) (Fig. 2). In the non-amended sediment
spiked with PHE, frapid quickly decreased and approached zero after
desorption for just 1 or 2 h (Fig. 2a). The decrease of fslow was more

Fig. 2. Relative fractions of rapid, slow, and very slow desorption pools after desorption for different time intervals (calculated from parameters in Table 1). (a) Non-
amended sediment spiked with phenanthrene; (b) charcoal-amended sediment spiked with phenanthrene; (c) non-amended sediment spiked with permethrin; and (d)
charcoal-amended sediment spiked with permethrin.
 X. Cui et al. / Aquatic Toxicology 98 (2010) 83–90 87

gradual, although it too reached zero following 96 h desorption.

Concurrently, fvslow increased with the desorption interval, and
accounted for essentially all the remaining chemical after 96 h des-
orption. A similar pattern was observed in the charcoal-amended
sediment for PHE, except that the overall frapid and fslow values
were smaller than those in the non-amended sediment at corre-
sponding desorption intervals (Fig. 2a and b). Compared to PHE,
the decrease of frapid and fslow in sediments with or without char-
coal amendment, or the increase in fvslow , was more gradual. In the
charcoal-amended sediment, fslow (0.22) was significantly greater
than zero even after 312 h of desorption, suggesting resistance in
PM desorption caused by addition of charcoal (Fig. 2c and d). The
charcoal amendment and sequential desorption scheme together
generated a large number of sediments that had different relative
compositions of rapid, slow, and very slow desorption pools, or sets
of frapid , fslow and fvslow values.

3.2. Bioaccumulation of PM and PHE by C. tentans

C. tentans were exposed to the above derived sediments to

obtain BSAFs for both PHE and PM (Fig. 3). The body residues and
chemical concentrations in sediment which were used to calculate
BSAFs are listed in Supporting Information Fig. S2. For all treat-
ments, as the sediments were increasingly desorbed, or frapid was
depleted, the BSAF values rapidly decreased. The decline in BSAF
followed approximately first-order decay kinetics. The decrease of
BSAF was considerably faster for PHE than for PM in both charcoal
amended and non-amended sediments, with the half-reduction
time being 7.7–9.5 h for PHE, and 34.7–69.3 h for PM. However, it is
important to note that the BSAF values did not decrease to zero even
when frapid became zero. For instance, after the 6 h desorption, frapid
was close to zero for the PHE spiked sediment (Fig. 2a), but the esti-
mated BSAF value was 5.88 ± 0.52, which was 47% of that measured
in the same sediment before desorption (Fig. 3a). In the charcoal
spiked sediment, frapid was effectively zero when the desorption
interval was ≥12 h (Fig. 2b), but the BSAF value was 2.91 ± 0.21 at
0 h and decreased to 1.13 ± 0.19 at 12 h (Fig. 3a). A similar pattern
was also observed for PM in both sediments. The frapid approached
zero after 48 h extraction for both sediments (Fig. 2a and b), but
the observed BSAF values were 0.34 ± 0.15 and 0.53 ± 0.09, equal-
Fig. 3. Biota-sediment accumulation factors (BSAFs) of phenanthrene (a) and
ing 18 and 25%, respectively, of those measured before desorption
permethrin (b) for C. tentans in non-amended sediment (shaded bar) and charcoal-
(Fig. 3b). As the body residue was determined after excluding the amended sediment (empty bar) after desorption for different time intervals.
gut content, the above observations clearly suggested that C. tentans Different letters on the bar for the same one time interval mean significant difference
was able to access fslow and fvslow . (p < 0.05) between the two treatments.
The addition of charcoal resulted in significant reductions
(p < 0.05) in BSAF values for PHE for all the desorption intervals.
Data from all four treatments were closely described by the model,
In the sediment without undergoing desorption, addition of char-
with p ≤0.001, and the independent variables explaining 94–99%
coal caused about 72% reduction in BSAF for PHE (Fig. 3a). On the
of the regression variations.
average, BSAF values in the charcoal-amended sediment were 19%
In the non-amended sediment, the mean BSAFslow of PHE was
of those in the non-amended sediment. In previous studies, amend-
17.2, which was 72.9% of that for BSAFrapid (23.6), suggesting
ment of activated carbon at 3% decreased the bioaccumulation of
reduced but nevertheless significant bioavailability of the slow des-
10 PAHs in Nereis diversicolor by 84% (Cornelissen et al., 2006;
orption fraction (Table 2). Both BSAFrapid and BSAFslow decreased in
Zimmerman et al., 2004). Unlike PHE, the BSAF values of PM after
the charcoal-amended sediment as compared to the non-amended
addition of charcoal were similar to those without charcoal amend-
ment for 8 out of the 12 desorption intervals, while the remaining
desorption intervals showed either slightly elevated or depressed Table 2
BSAFs (Fig. 3b). The difference between the two groups of BSAF Parameters from fit of the measured biota-sediment accumulation factor (BSAF)
values to the multivariate model (Eq. (4)) for bioaccumulation of phenanthrene and
values was not significantly different from 0 (t-test, p = 0.36). This
permethrin by C. tentans in non-amended and charcoal-amended sediments.
observation was consistent with the fact that charcoal had limited
effect on desorption of PM from the sediment phase. Sediment BSAFrapid BSAFslow BSAFvslow R2 P

Phenanthrene
3.3. Contributions from different desorption fractions Non-amended 23.6 ± 3.9 17.2 ± 1.3 1.4 ± 0.4 0.94 <0.001
Charcoal-amended 14.6 ± 0.9 7.4 ± 0.5 0.2 ± 0.1 0.99 <0.001

The calculated frapid , fslow , and fvslow at various desorption inter- Permethrin
vals and the measured BSAF values were fitted into Eq. (4) to derive Non-amended 7.2 ± 1.1 4.4 ± 0.8 0 0.96 <0.001
Charcoal-amended 10.9 ± 0.7 2.8 ± 0.3 0 0.98 <0.001
BSAFs expressed on the basis of the three desorption pools (Table 2).
88 X. Cui et al. / Aquatic Toxicology 98 (2010) 83–90

In the two treatments spiked with PM, the relative contributions

from the rapid and slow desorption pools showed a generally sim-
ilar trend to that of PHE in the same sediments (Fig. 5). However,
the contribution from frapid decreased considerably more gradually
compared to PHE, and the contribution did not reach zero till after
the 96 h interval. Again, as frapid was depleted by desorption, the
contribution from fslow increased, reaching the maximum value of
about 100% for both sediments after 144 h desorption.
It is important to note that the overwhelming contributions
from fvslow after prolonged desorption as observed for PHE was a
result of chemical depletion in the other two desorption fractions.
The absolute availability of these chemicals in the different des-
orption pools was more directly reflected in the respective BSAF
values (Table 2). Therefore, PHE or PM in the very slow desorption
pool was poorly available to C. tentans, and the limited availability
led to the overall small BSAFs observed after prolonged desorption.
Results from this study provided direct experimental evidence
suggesting that sediment-sorbed hydrophobic chemicals such as
PHE and PM in the kinetically slow desorption fraction (fslow ) were
partially bioavailable to C. tentans, while the fraction highly resis-
tant to desorption was essentially unavailable. The availability of
PHE or PM in fslow was consistent with the partial availability of
slow desorption HOCs observed for benthic organisms such as
Tubifex tubifex Müller (Kraaij et al., 2002), C. tentans (Leppänen et al.,

Fig. 4. Contributions from rapid (), slow (䊉), and very slow desorption fractions
() to phenanthrene accumulation in C. tentans. (a) Non-amended sediment and (b)
charcoal-amended sediment.

sediment, but BSAFslow still equaled to 50.8% of BSAFrapid . For PM,

BSAFslow in the non-amended sediment was 60.7% of BSAFrapid ,
which decreased to 25.3% in the charcoal-amended sediment
(Table 2). In contrast, BSAFvslow was either zero (for PM), or very
small (for PHE), suggesting much reduced availability of the very
slow sorption fraction.
Based on the estimated BSAFrapid, BSAFslow , and BSAFvslow
values, and the calculated frapid , fslow , and fvslow , the relative con-
tributions from the different desorption pools were estimated for
all desorption intervals (Figs. 4 and 5). In the non-amended sedi-
ment, Frapid (prior to desorption) contributed 49.2% to the overall
bioaccumulation of PHE in C. tentans. In the charcoal-amended
sediment, the initial contribution from Frapid was 60.9% (Fig. 4).
In both sediments, the contribution from frapid rapidly decreased
as the desorption interval increased (Fig. 4). For instance, after
24 h desorption, contribution from frapid was not significantly dif-
ferent from zero. As the contribution from frapid diminished, the
relative contribution from fslow concurrently increased, reaching
a maximum of 90.6% after 2 h desorption. However, as the sedi-
ment samples were further desorbed, the contribution from this
pool also declined, reaching zero after 192 h desorption. As frapid
and fslow were both depleted by prolonged desorption, the con-
Fig. 5. Contributions from rapid () and slow desorption fractions (䊉) to permethrin
tribution from fvslow increased rapidly and reached the maximum accumulation in C. tentans. (a) Non-amended sediment and (b) charcoal-amended
value of about 100% after the 192 h desorption interval. sediment.
 X. Cui et al. / Aquatic Toxicology 98 (2010) 83–90
2003), and Lumbriculus variegatus (Kukkonen et al., 2004; Leppänen
et al., 2003). However, causes for the uptake of sorbed HOCs in
sediment by benthic organisms are not well understood. One pos-
sible explanation is that desorption from the slow desorption pool
may slowly replenish the depleted pore water, while the release of
tightly bound HOCs may be kinetically too slow to contribute signif-
icantly to bioaccumulation (Landrum and Robbins, 1990; Sijm et al.,
2000). It is also possible that some biological factors, such as inges-
tion of sediment particles and enhanced dissolution or desorption
caused by the gut fluid and the consequent transit through the gut
(Forbes et al., 1998; Gobas et al., 1993; Leppänen and Kukkonen,
2000; Timmermann and Andersen, 2003), resulted in the accumu-
lation of sorbed PHE or PM by C. tentans. For instance, it was found
that 32–68% of pyrene accumulation in Lumbriculus variegatus came
from utilization of sediment-associated pyrene (Conrad et al., 2002;
Leppänen and Kukkonen, 1998). Conrad et al. (2002) showed that
the “non-labile” fraction of pyrene (slow desorption fraction) after
1–3 months of aging was still available through digestive uptake in
Lumbriculus variegatus. In some recent studies, PAHs, such as PHE,
that were sorbed by strong sorbents (e.g., activated charcoal) were
found to be bioavailable, and the uptake was attributed to the direct
use of solid particles (Rhodes et al., 2008).
Therefore, it may be inaccurate to define HOCs in the slow des-
orption pool as either bioavailable or unavailable, given that this
fraction is partially bioavailable, although the very slow desorption
pool may be effectively unavailable. Understanding the makeup of
the sorbed pool, or the desorption kinetics, would then be criti-
cal for predicting the bioavailability of sediment-sorbed HOCs. In
addition, it would be of great value to devise less time consuming
methods than the tedious sequential desorption approach, and also
to evaluate the effect of environmental factors and conditions, such
as aging and organic carbon compositions, on the relative makeup
of the different desorption pools and hence bioavailability of sorbed
HOCs.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at doi:10.1016/j.aquatox.2010.01.016.
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