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Bioaccumulation: aquatic / sediment

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Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)
Principles of method if other than guideline:
The study includes chemical analysis, distribution, metabolism and excretion of PAH in fish. Eight PAH were investigated (two- to four-ring PAH including alkyl substituted substances). Metabolites (hydroxylated products) were determined in gallbladder/bile.
The test substance covered in this study record is PHENANTHRENE:
GLP compliance:
not specified
Specific details on test material used for the study:
Eight PAH were tested in this study comprising naphthalene, phenanthrene, and pyrene (two- to four-ring PAH) as well as alkyl substituted naphthalenes and phenanthrenes. Two compounds among them are relevant as constituents of anthracene oil (AOL). These are positioned at the top of following table. The substance phenanthrene is covered in this study record.

The following PAH are included in this study. Compounds were purchased from Chiron, Trondheim, Norway. Purity is not specified in the publication. Test substances were applied dissolved in marine water in this bioaccumulation study. The final concentration in the test medium (low and high concentration level, measured values) is also presented in the table.

Exposure concentration [µg/L] (measured value ± standard deviation)

low level (n = 17) high level (n = 15)
================================================================
Phenanthrene 0.12 ± 0.03 1.12 ± 0.32
Pyrene 0.012 ± 0.002 0.10 ± 0.02
--------------------------------------------------------------------------------------------------------------------------------
Naphthalene 1.34 ± 0.37 12.5 ± 3.8
2-Methylnaphthalene 2.46 ± 0.68 23.63 ± 7.84
1,3-Dimethylnaphthalene 2.74 ± 0.73 26.67 ± 9.19
2-Isopropylnaphthalene 0.70 ± 0.18 6.40 ± 2.09
9-Methylphenanthrene 0.14 ± 0.035 1.35 ± 0.36
9-Ethylphenanthrene 0.062 ± 0.015 0.51 ± 0.11
================================================================
Total PAH 7.57 ± 1.98 72.31 ± 23.37
Radiolabelling:
no
Details on sampling:
- Sampling intervals/frequency for test organisms (including bile): during exposure at day 0, 0.17, 0.42, 1, 2, 4, 7, 14, 21, 28, 36 and during depuration at day 36.5 (0.5), 37 (1), 38 (2), 40 (4), and 44 (8)
- Sampling intervals/frequency for test medium samples: during exposure at day 0, 0.17, 0.42, 1, 2, 4, 7, 21, 28, and 36
- Sample storage conditions before analysis: water samples were acidified (pH < 2) for stabilisation and prepared for analysis within 48 hrs; fish samples were homogenised (cut into pieces) and stored in glass containers at -80 °C; gallbladders as a whole were removed from the fish and stored at -80 °C.
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods):
- Seawater: samples were collected from the middle of the exposure chambers into 1-L glass bottles containing hydrochloric acid by use of a Teflon siphon connected to a piece of glass tubing. Hydrochloric acid was used in order to stabilise the sample at a low pH (< 2).
- Fish: the weight and length of individual fish were recorded before fish were killed by a blow to the head. The fish were dissected with whole gallbladders removed and stored at -80 °C until analysis.
Vehicle:
yes
Remarks:
acetone
Details on preparation of test solutions, spiked fish food or sediment:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a PAH stock solution in acetone (11.894 mg/L) was continuously introduced into a mixing container via a syringe pump (ISCO, model 260D; HOUM AS, Oslo, Norway), where it was diluted by a factor of 40,000 with seawater. Experimental exposure concentrations were obtained by further dilution of this stock solution with seawater. Nominal concentrations (total PAHs) in the low- and high-exposure chambers were 11.1 and 100 µg/L respectively.
- Controls: pure seawater and 25 µL acetone/L seawater (solvent control, dilution 1:40,000)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): ca. 8.3 and 0.93 µL/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no
Test organisms (species):
Cyprinodon variegatus
Details on test organisms:
TEST ORGANISM
- Common name: sheepshead minnow
- Strain:
- Source: commercial, Sea Plantations, Salem, MA, USA and Aquatic Research Organisms, Hampton, NH, USA
- Age at study initiation (mean and range, SD): adult
- Exposed fish
- Length at sampling time (lenght definition, mean, range and SD): 4.7 ± 0.8 cm
- Weight at sampling time (mean and range, SD): 2.47 ± 1.23 g
- Control fish
- Length at sampling timed (lenght definition, mean, range and SD): 4.7 ± 0.5 cm
- Weight at sampling time (mean and range, SD): 2.33 ± 0.90 g
- Feeding during test
- Food type: raw peeled shrimp
- Amount: no data
- Frequency: daily

ACCLIMATION
- Acclimation period: at least four weeks
- Acclimation conditions (same as test or not): yes; 25 °C and 3.4 % salinity
- Type and amount of food: commercial fish pellets (Nutra Svev, Skretting, Stavanger, Norway) and raw peeled shrimp
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no data
Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: marine
Total exposure / uptake duration:
36 d
Total depuration duration:
8 d
Test temperature:
25 °C
Salinity:
3.4 %
Details on test conditions:
TEST SYSTEM (according to Bechmann e al. (2000). DREAM ― Reproduction effects of a PAH mixture on sheepshead minnow (Cyprinodon variegatus). Report am-2000/015. Akvamiljø AS, Randaberg, Norway)
- Test vessel: not specified
- Aeration: not specified
- Renewal rate of test solution (frequency/flow rate): not specified
- No. of organisms per vessel: 79 females and 65 males
- No. of vessels per concentration (replicates): 1
- No. of vessels per control / vehicle control (replicates): two for controls, one for solvent control (70 females and 45 males each)
- Biomass loading rate: not specified

TEST MEDIUM / WATER PARAMETERS
- no further information
Nominal and measured concentrations:
Sum of PAH
Measured concentrations: Nominal concentrations:
low level: 7.57 µg/L; high level: 72.31 µg/L low level: 11.1 µg/L; high level: 100 µg/L

Phenanthrene
Measured concentrations:
low level: 0.12 ± 0.03 µg/L (n = 17); high level: 1.12 ± 0.32 µg/L (n = 15)
Reference substance (positive control):
no
Lipid content:
ca. 10.3 %
Time point:
end of exposure
Remarks on result:
other: estimated from ratios of BCF(lipid weight) / BCF(wet weight) (kinetic BCF values)
Key result
Conc. / dose:
0.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
393 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance phenanthrene: lipid normalised BCF - highest value at the low exposure level (0.12 µg/L)
Key result
Conc. / dose:
1.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
1 082 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance phenanthrene: lipid normalised BCF - highest value at the high exposure level (1.12 µg/L)
Key result
Conc. / dose:
0.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
810 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance phenanthrene - highest value at the low exposure level
Key result
Conc. / dose:
0.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
700 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
steady state
Remarks on result:
other: test substance phenanthrene: BCF = mean of determinations at sampling days 4, 7, and 30
Remarks:
low exposure level
Key result
Conc. / dose:
1.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
2 229 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance phenanthrene - highest value at the high exposure level
Key result
Conc. / dose:
1.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
1 623 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
steady state
Remarks on result:
other: test substance phenanthrene: BCF = mean of determinations at sampling days 4, 7, and 30
Remarks:
high exposure level
Key result
Elimination:
yes
Parameter:
other: T95 = Time interval for 95 % excretion
Depuration time (DT):
3.6 d
Remarks on result:
other: low exposure level
Remarks:
reported in the publication
Key result
Elimination:
yes
Parameter:
DT50
Depuration time (DT):
0.825 d
Remarks on result:
other: low exposure level
Remarks:
value calculated from kinetic data reported in the publication
Key result
Rate constant:
other: uptake rate constant k1 (g water/g fish/d)
Value:
680
Remarks on result:
other: SE: ± 89; low exposure level
Remarks:
curve fitting not significant at p < 0.05
Key result
Rate constant:
overall depuration rate constant (d-1)
Value:
0.84
Remarks on result:
other: SE: ± 0.08; low exposure level
Key result
Rate constant:
other: uptake rate constant k1 (g water/g fish/d)
Value:
1 783
Remarks on result:
other: SE: ± 104; high exposure level
Key result
Rate constant:
overall depuration rate constant (d-1)
Value:
0.8
Remarks on result:
other: SE: ± 0.06; high exposure level
Metabolites:
hydroxy compounds
Reported statistics:
All data analysis was performed by use of JMP software (Vers 3.2.2; SAS Institute, Cary, NC, USA). Comparison of the means of data sets was investigated by the nonparametric Kruskal-Wallis test, with further comparison by Tukey-Kramer honestly significant difference (multiple comparisons). The level of statistical significance was set to p < 0.05. Quantile box-and-whisker plots were drawn for the data sets to illustrate the interquartile range, median, data distribution, and possible outliers.
Validity criteria fulfilled:
not specified
Conclusions:
In this study, phenanthrene shows low bioaccumulating potential due to very efficient metabolism and high excretion. The BCF based on the standard calculation basis (steady state) was below 2000 (=>1623). When normalised to a standard lipid content of 5 %, the highest BCF determined (2229, kinetic BCF, high exposure level) is 1082.
Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The source test material is an artificial mixture of unsubstituted and alkyl substituted polycyclic aromatic hydrocarbons with two to four aromatic rings purchased from commercial sources. One constituent of the test mixture is phenanthrene. Phenanthrene consists of three aromatic rings and is besides others a major constituent of the target substance. The target substance anthracene oil (benzo[a]pyrene < 50 ppm, AOL) is composed of a broad range of PAH predominantly consisting of two up to four aromatic rings.
The aquatic bioaccumulation potential of anthracene oil will be characterised by the range of PAH that constitute its composition. Therefore, results obtained from an aquatic bioaccumulation study with the AOL constituent phenanthrene can be used in order to characterise the bioaccumulation potential of anthracene oil itself.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source test material covered in this study record is the substance phenanthrene. It was applied in the source study as a component in a hand-made mixture of PAH consisting of two-, three- and four-ring PAH some of them alkylated. The test substances were purchased from a commercial source (Trondheim, Norway). The analytical purity of the test materials are not reported. But it is assumed that the purity of the commercial products is high. Furthermore, the test results reported are allocated to individual test substances (analytical determination of individual substance). Thus, the study results are unequivocally related to effects caused by phenanthrene and are considered to be valid for the substance phenanthrene.
The target material anthracene oil is a UVCB substance produced by the distillation of coal tars extracting the approximate distillation range from ca. 300 °C to 400 °C. 10 % to 95 % of the total product distil over between ca. 300 °C and 375 °C. The substance is a brown pasty or liquid material consisting of a complex and within limits variable combination of polycyclic aromatic hydrocarbons. PAH contained in AOL range from naphthalene up to pyrene and benzofluorenes. Two- and three-ring aromatics amount to about 50 % (typical concentration). Main constituent is phenanthrene present in a typical concentration of ca. 25 to 31 % (composite sample 7). PAH with four rings accumulate to about 10 %.

3. ANALOGUE APPROACH JUSTIFICATION
Properties of the target substance anthracene oil relevant for environmental distribution will be determined by the properties of the PAH that are constituents of anthracene oil. Under environmental conditions or during processing of the target substance, environmentally available or volatile/water soluble components of anthracene oil can be released. These will be PAH (mainly compounds consisting of two to four fused rings). In combination, they will specify the environmental properties and in this case the aquatic bioconcentration of anthracene oil as a whole. In addition, phenanthrene is considered to adequately represent the overall bioconcentration potential of AOL. It is one of the most abundant components in AOL with the ability for bioaccumulation characteristic for AOL. Therefore, phenanthrene is selected as marker substance to represent the aquatic properties of AOL. For these reasons, it is considered justified to use date determined for phenanthrene as constituent of anthracene oil to characterise environmental behaviour (aquatic bioaccumulation) of anthracene oil itself.
Reason / purpose for cross-reference:
read-across source
Principles of method if other than guideline:
Read-across to preceding entry:
Source test material: polycyclic aromatic hydrocarbons (PAH), generic mixture - substance PHENANTHRENE;
Reference: Jonsson et al. 2004
Lipid content:
ca. 10.3 %
Time point:
end of exposure
Remarks on result:
other: estimated from ratios of BCF(lipid weight) / BCF(wet weight) (kinetic BCF values)
Key result
Conc. / dose:
0.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
392 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance phenanthrene: lipid normalised BCF - highest value at the low concentration level (0.12 µg/L) (original BCF value: 810); the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Remarks:
for originally measured values (kinetic and steady state) see source entry (Jonsson et al. 2014, PAH generic mixture - substance phenanthrene
Key result
Conc. / dose:
1.12 µg/L
Temp.:
25 °C
Type:
BCF
Value:
1 082 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance phenanthrene: lipid normalised BCF - highest value at the high concentration level (1.12 µg/L) (original BCF value 2229); the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Remarks:
for originally measured values (kinetic and steady state) see source entry (Jonsson et al. 2014, PAH generic mixture - substance phenanthrene
Key result
Elimination:
yes
Parameter:
other: T95 = Time interval for 95 % excretion
Depuration time (DT):
3.6 d
Remarks on result:
other: low exposure level; reported in the publication
Remarks:
the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Key result
Elimination:
yes
Parameter:
DT50
Depuration time (DT):
0.825 d
Remarks on result:
other: low exposure level; value calculated from kinetic data reported in the publication
Remarks:
the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Details on kinetic parameters:
for individual uptake and depuration rate constants see source study record
Conclusions:
In this study, phenanthrene shows low bioaccumulating potential due to very efficient metabolism and high excretion. The BCF based on the standard calculation basis (steady state) was below 2000 (=>1623). When normalised to a standard lipid content of 5 %, the highest BCF determined (2229, kinetic BCF, high exposure level) is 1082.
The study results of the source substance phenanthrene, a major constituent of the target substance anthracene oil, are adopted as weight of evidence for the target substance.
Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 305 (Bioconcentration: Flow-through Fish Test)
Principles of method if other than guideline:
The study includes chemical analysis, distribution, metabolism and excretion of PAH in fish. Eight PAH were investigated (two- to four-ring PAH including alkyl substituted substances). Metabolites (hydroxylated products) were determined in gallbladder/bile.
The test substance covered in this endpoint study record is PYRENE.
GLP compliance:
not specified
Specific details on test material used for the study:
Eight PAH were tested in this study comprising naphthalene, phenanthrene, and pyrene (two- to four-ring PAH) as well as alkyl substituted naphthalenes and phenanthrenes. Two compounds among them are relevant as constituents of anthracene oil (AOL). These are positioned at the top of following table. The substance pyrene is covered in this study record.

The following PAH are included in this study. Compounds were purchased from Chiron, Trondheim, Norway. Purity is not specified in the publication. Test substances were applied dissolved in marine water in this bioaccumulation study. The final concentration in the test medium (low and high concentration level, measured values) is also presented in the table.

Exposure concentration [µg/L] (measured value ± standard deviation)

low level (n = 17) high level (n = 15)
================================================================
Phenanthrene 0.12 ± 0.03 1.12 ± 0.32
Pyrene 0.012 ± 0.002 0.10 ± 0.02
--------------------------------------------------------------------------------------------------------------------------------
Naphthalene 1.34 ± 0.37 12.5 ± 3.8
2-Methylnaphthalene 2.46 ± 0.68 23.63 ± 7.84
1,3-Dimethylnaphthalene 2.74 ± 0.73 26.67 ± 9.19
2-Isopropylnaphthalene 0.70 ± 0.18 6.40 ± 2.09
9-Methylphenanthrene 0.14 ± 0.035 1.35 ± 0.36
9-Ethylphenanthrene 0.062 ± 0.015 0.51 ± 0.11
================================================================
Total PAH 7.57 ± 1.98 72.31 ± 23.37
Radiolabelling:
no
Details on sampling:
- Sampling intervals/frequency for test organisms (including bile): during exposure at day 0, 0.17, 0.42, 1, 2, 4, 7, 14, 21, 28, 36 and during depuration at day 36.5 (0.5), 37 (1), 38 (2), 40 (4), and 44 (8)
- Sampling intervals/frequency for test medium samples: during exposure at day 0, 0.17, 0.42, 1, 2, 4, 7, 21, 28, and 36
- Sample storage conditions before analysis: water samples were acidified (pH < 2) for stabilisation and prepared for analysis within 48 hrs; fish samples were homogenised (cut into pieces) and stored in glass containers at -80 °C; gallbladders as a whole were removed from the fish and stored at -80 °C.
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods):
- Seawater: samples were collected from the middle of the exposure chambers into 1-L glass bottles containing hydrochloric acid by use of a Teflon siphon connected to a piece of glass tubing. Hydrochloric acid was used in order to stabilise the sample at a low pH (< 2).
- Fish: the weight and length of individual fish were recorded before fish were killed by a blow to the head. The fish were dissected with whole gallbladders removed and stored at -80 °C until analysis.
Vehicle:
yes
Remarks:
acetone
Details on preparation of test solutions, spiked fish food or sediment:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a PAH stock solution in acetone (11.894 mg/L) was continuously introduced into a mixing container via a syringe pump (ISCO, model 260D; HOUM AS, Oslo, Norway), where it was diluted by a factor of 40,000 with seawater. Experimental exposure concentrations were obtained by further dilution of this stock solution with seawater. Nominal concentrations (total PAHs) in the low- and high-exposure chambers were 11.1 and 100 µg/L respectively.
- Controls: pure seawater and 25 µL acetone/L seawater (solvent control, dilution 1:40,000)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): ca. 8.3 and 0.93 µL/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no
Test organisms (species):
Cyprinodon variegatus
Details on test organisms:
TEST ORGANISM
- Common name: sheepshead minnow
- Strain:
- Source: commercial, Sea Plantations, Salem, MA, USA and Aquatic Research Organisms, Hampton, NH, USA
- Age at study initiation (mean and range, SD): adult
- Exposed fish
- Length at sampling time (lenght definition, mean, range and SD): 4.7 ± 0.8 cm
- Weight at sampling time (mean and range, SD): 2.47 ± 1.23 g
- Control fish
- Length at sampling timed (lenght definition, mean, range and SD): 4.7 ± 0.5 cm
- Weight at sampling time (mean and range, SD): 2.33 ± 0.90 g
- Feeding during test
- Food type: raw peeled shrimp
- Amount: no data
- Frequency: daily

ACCLIMATION
- Acclimation period: at least four weeks
- Acclimation conditions (same as test or not): yes; 25 °C and 3.4 % salinity
- Type and amount of food: commercial fish pellets (Nutra Svev, Skretting, Stavanger, Norway) and raw peeled shrimp
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no data
Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: marine
Total exposure / uptake duration:
36 d
Total depuration duration:
8 d
Test temperature:
25 °C
Salinity:
3.4 %
Details on test conditions:
TEST SYSTEM (according to Bechmann e al. (2000). DREAM ― Reproduction effects of a PAH mixture on sheepshead minnow (Cyprinodon variegatus). Report am-2000/015. Akvamiljø AS, Randaberg, Norway)
- Test vessel: not specified
- Aeration: not specified
- Renewal rate of test solution (frequency/flow rate): not specified
- No. of organisms per vessel: 79 females and 65 males
- No. of vessels per concentration (replicates): 1
- No. of vessels per control / vehicle control (replicates): two for controls, one for solvent control (70 females and 45 males each)
- Biomass loading rate: not specified

TEST MEDIUM / WATER PARAMETERS
- no further information
Nominal and measured concentrations:
Sum of PAH
Measured concentrations: Nominal concentrations:
low level: 7.57 µg/L; high level: 72.31 µg/L low level: 11.1 µg/L; high level: 100 µg/L

Pyrene
Measured concentrations:
low level: 0.012 ± 0.002 µg/L (n = 17); high level: 0.10 ± 0.02 µg/L (n = 15)
Reference substance (positive control):
no
Lipid content:
ca. 10.3 %
Time point:
end of exposure
Remarks on result:
other: estimated from ratios of BCF(lipid weight) / BCF(wet weight) (kinetic BCF values)
Key result
Conc. / dose:
0.012 µg/L
Temp.:
25 °C
Type:
BCF
Value:
70.4 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance pyrene: lipid normalised BCF - highest value at the low exposure level (0.012 µg/L)
Key result
Conc. / dose:
0.1 µg/L
Temp.:
25 °C
Type:
BCF
Value:
47 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance pyrene: lipid normalised BCF - highest value at the high exposure level (0.10 µg/L)
Key result
Conc. / dose:
0.012 µg/L
Temp.:
25 °C
Type:
BCF
Value:
145 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance pyrene - highest value at the low exposure level
Remarks:
BCF was calculated from non-significant uptake rate constant (k1)
Key result
Conc. / dose:
0.012 µg/L
Temp.:
25 °C
Type:
BCF
Value:
50 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
steady state
Remarks on result:
other: test substance pyrene: BCF = mean of determinations at sampling days 4, 7, and 30
Remarks:
low exposure level
Key result
Conc. / dose:
0.1 µg/L
Temp.:
25 °C
Type:
BCF
Value:
97 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance pyrene - highest value at the high exposure level
Remarks:
BCF was calculated from non-significant uptake rate constant (k1)
Key result
Conc. / dose:
0.1 µg/L
Temp.:
25 °C
Type:
BCF
Value:
53 dimensionless
Basis:
whole body w.w.
Time of plateau:
4 d
Calculation basis:
steady state
Remarks on result:
other: test substance pyrene: BCF = mean of determinations at sampling days 4, 7, and 30
Remarks:
high exposure level
Key result
Elimination:
yes
Parameter:
other: T95 = Time interval for 95 % excretion
Depuration time (DT):
3.4 d
Remarks on result:
other: low exposure level
Remarks:
reported in the publication
Key result
Elimination:
yes
Parameter:
DT50
Depuration time (DT):
0.779 d
Remarks on result:
other: low exposure level
Remarks:
value calculated from kinetic data reported in the publication
Key result
Elimination:
yes
Parameter:
other: T95 = Time interval for 95 % excretion
Depuration time (DT):
2.5 d
Remarks on result:
other: high exposure level
Remarks:
reported in the publication
Key result
Elimination:
yes
Parameter:
DT50
Depuration time (DT):
0.583 d
Remarks on result:
other: high exposure level
Remarks:
value calculated from kinetic data reported in the publication
Key result
Rate constant:
other: uptake rate constant k1 (g water/g fish/d)
Value:
129
Remarks on result:
other: SE: ± 28; low exposure level
Remarks:
curve fitting not significant at p < 0.05
Key result
Rate constant:
overall depuration rate constant (d-1)
Value:
0.89
Remarks on result:
other: SE: ± 0.22; low exposure level
Key result
Rate constant:
other: uptake rate constant k1 (g water/g fish/d)
Value:
116
Remarks on result:
other: SE: ± 15; high exposure level
Remarks:
curve fitting not significant at p < 0.05
Key result
Rate constant:
overall depuration rate constant (d-1)
Value:
1.19
Remarks on result:
other: SE: ± 0.28; high exposure level
Metabolites:
hydroxy compounds
Reported statistics:
All data analysis was performed by use of JMP software (Vers 3.2.2; SAS Institute, Cary, NC, USA). Comparison of the means of data sets was investigated by the nonparametric Kruskal-Wallis test, with further comparison by Tukey-Kramer honestly significant difference (multiple comparisons). The level of statistical significance was set to p < 0.05. Quantile box-and-whisker plots were drawn for the data sets to illustrate the interquartile range, median, data distribution, and possible outliers.
Validity criteria fulfilled:
not specified
Conclusions:
In this study, pyrene showed a very low bioaccumulation potential due to very efficient metabolism and high excretion. Steady state BCF values for pyrene were 50 and 53 (low and high exposure concentration, respectively). The somewhat higher kinetically derived BCF values (145 and 97) decreased to values of 70 and 41 when normalised to a standard lipid content of 5 %.
Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The source test material is an artificial mixture of unsubstituted and alkyl substituted polycyclic aromatic hydrocarbons with two to four aromatic rings purchased from commercial sources. One constituent of the test mixture is pyrene. Pyrene consists of four aromatic rings and is besides others a main constituent of the target substance. The target substance anthracene oil (benzo[a]pyrene < 50 ppm, AOL) is composed of a broad range of PAH predominantly consisting of two up to four aromatic rings.
The aquatic bioaccumulation potential of anthracene oil will be characterised by the range of PAH that constitute its composition. Therefore, results obtained from an aquatic bioaccumulation study with the AOL constituent pyrene can be used in order to characterise the bioaccumulation potential of anthracene oil itself.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source test material covered in this study record is the substance pyrene. It was applied in the source study as a component in a hand-made mixture of PAH consisting of two-, three- and four-ring PAH some of them alkylated. The test substances were purchased from a commercial source (Trondheim, Norway). The analytical purity of the test materials are not reported. But it is assumed that the purity of the commercial products is high. Furthermore, the test results reported are allocated to individual test substances (analytical determination of individual substance). Thus, the study results are unequivocally related to effects caused by pyrene and are considered to be valid for the substance pyrene.
The target material anthracene oil is a UVCB substance produced by the distillation of coal tars extracting the approximate distillation range from ca. 300 °C to 400 °C. 10 % to 95 % of the total product distil over between ca. 300 °C and 375 °C. The substance is a brown pasty or liquid material consisting of a complex and within limits variable combination of polycyclic aromatic hydrocarbons. PAH contained in AOL range from naphthalene up to pyrene and benzofluorenes. Two- and three-ring aromatics amount to about 50 % (typical concentration). Main constituent is phenanthrene present in a typical concentration of ca. 25 to 31 % (composite sample 7). PAH with four rings accumulate to about 10 %.

3. ANALOGUE APPROACH JUSTIFICATION
Properties of the target substance anthracene oil relevant for environmental distribution will be determined by the properties of the PAH that are constituents of anthracene oil. Under environmental conditions or during processing of the target substance, environmentally available or volatile/water soluble components of anthracene oil can be released. These will be PAH (mainly compounds consisting of two to four fused rings). In combination, they will specify the environmental properties and in this case the aquatic bioconcentration of anthracene oil as a whole. Therefore, it is considered justified to use date determined for pyrene as constituent of anthracene oil to characterise environmental behaviour (aquatic bioaccumulation) of anthracene oil itself.
Reason / purpose for cross-reference:
read-across source
Principles of method if other than guideline:
Read-across to preceding entry:
Source test material: polycyclic aromatic hydrocarbons (PAH), generic mixture - substance PYRENE;
Reference: Jonsson et al. 2004
Lipid content:
10.3 %
Time point:
end of exposure
Remarks on result:
other: estimated from ratios of BCF(lipid weight) / BCF(wet weight) (kinetic BCF values)
Key result
Conc. / dose:
0.012 µg/L
Temp.:
25 °C
Type:
BCF
Value:
70.4 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance pyrene: lipid normalised BCF - highest value at the low exposure level (0.012 µg/L) (original BCF value: 145); the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Remarks:
for originally measured values (kinetic and steady state) see source entry (Jonsson et al. 2014, PAH generic mixture - substance pyrene
Key result
Conc. / dose:
0.1 µg/L
Temp.:
25 °C
Type:
BCF
Value:
47 dimensionless
Basis:
normalised lipid fraction
Remarks:
from 10.3 % lipid extrapolated to 5 % (standard)
Time of plateau:
4 d
Calculation basis:
kinetic
Remarks on result:
other: test substance pyrene: lipid normalised BCF - highest value at the high exposure level (0.10 µg/L) (original BCF value: 97); the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Remarks:
for originally measured values (kinetic and steady state) see source entry (Jonsson et al. 2014, PAH generic mixture - substance pyrene
Key result
Elimination:
yes
Parameter:
other: T95 = Time interval for 95 % excretion
Depuration time (DT):
3.4 d
Remarks on result:
other: low exposure level; value reported in the publication
Remarks:
the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Key result
Elimination:
yes
Parameter:
DT50
Depuration time (DT):
0.779 d
Remarks on result:
other: low exposure level; value calculated from kinetic data reported in the publication
Remarks:
the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Key result
Elimination:
yes
Parameter:
other: T95 = Time interval for 95 % excretion
Depuration time (DT):
2.5 d
Remarks on result:
other: high exposure level; value reported in the publication
Remarks:
the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Key result
Elimination:
yes
Parameter:
DT50
Depuration time (DT):
0.583 d
Remarks on result:
other: high exposure level; value calculated from kinetic data reported in the publication
Remarks:
the test result of the source substance is adopted as weight of evidence for the target substance anthracene oil
Details on kinetic parameters:
for individual uptake and depuration rate constants see source study record
Conclusions:
In this study, the source substance pyrene showed a very low bioaccumulation potential in fish due to very efficient metabolism and high excretion. Steady state BCF values for pyrene were 50 and 53 (low and high exposure concentration, respectively). The somewhat higher kinetically derived BCF values (145 and 97) decreased to values of 70 and 41 when normalised to a standard lipid content of 5 %.
The study results of the source substance pyrene, a main constituent of the target substance anthracene oil, is adopted as weight of evidence for the target substance.
Endpoint:
bioaccumulation in aquatic species, other
Remarks:
various organisms of different trophic levels
Type of information:
experimental study
Remarks:
information comes from a comprehensive report summarising properties of a variety of polycyclic aromatic hydrocarbons
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Review, peer-reviewed data
Qualifier:
no guideline available
Principles of method if other than guideline:
Review on PAHs summarising data peer-reviewed by a group of international experts
GLP compliance:
not specified
Specific details on test material used for the study:
PAH consisting of two up to four aromatic ring systems are relevant components of anthracene oil. Their properties are used for the characterisation of the bioaccumulation potential of anthracene oil.
Test organisms (species):
other: various
Route of exposure:
other: various
Test type:
other: various
Water / sediment media type:
other: various
Key result
Remarks on result:
other: No bioaccumulating potential of PAH in fish and mammals, and hence in the food chain, due to effective metabolism and excretion in these species.

Bioaccumulation

Aquatic organisms that metabolise PAHs to little or no extent, such as algae, molluscs and the more primitive invertebrates (protozoans, porifers and cnidaria) accumulate high concentrations of PAHs, as would be expected from their log Kow values, whereas organisms that metabolise PAHs to a great extent, such as fish and higher invertebrates, accumulate little or no PAHs.

The concentration of PAHs in vegetation is generally considerably lower than that in soil, the bioaccumulation factors ranging from 0.0001 to 0.33 for BaP and from 0.001 to 0.18 for 17 other PAHs tested including PAH constituents of anthracene oil.

Biomagnification (the increase in concentration of a substance in animals in successive trophic levels of food chains) of PAHs has not been observed in aquatic systems and would not be expected to occur, because most organisms have a high biotransformation potential for PAHs. Organisms at higher trophic levels in food chains show the highest potential for biotransformation (WHO 1998).

[from reference Joint WHO/CONVENTION TAKS FORCE 2003, p.147]

Endpoint:
bioaccumulation in aquatic species, other
Remarks:
various species
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
The source test materials are individual polycyclic aromatic hydrocarbons ranging from two- to six-ring PAH. Depending on size/molecular weight and structure, physico-chemical and environmental properties can be different. The target substance anthracene oil (benzo[a]pyrene < 50 ppm, AOL) is as well composed of a broad range of PAH consisting of two to four aromatic rings.
The potential for bioaccumulation of anthracene oil in aquatic species will be characterised by the range of PAH that constitute its composition. Some of the source substances are major constituents of anthracene oil. Therefore, results obtained from aquatic bioaccumulation studies with these compounds can be used in order to characterise the bioaccumulation potential of anthracene oil.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
Source chemicals are individual PAH. These include major constituents of anthracene oil. The source reference is a comprehensive review compiled by technical experts that cover Long Range Transboundary Air Pollution. For this purpose, various properties among others bioaccumulation specific for individual substances have been considered. Individual PAH test materials are not specified, but due to the review process by experts, results are considered to be valid and specific for the individual test substances as well as for PAH as a chemical class of substances.
The target material anthracene oil (AOL) is a UVCB substance produced by the distillation of coal tars extracting the approximate distillation range from ca. 300 °C to 400 °C. This distillation range excludes mostly low molecular aromatic hydrocarbons (especially one-ring and to a lower extent two-ring aromatics) as well as polycyclic aromatic hydrocarbons composed of more than four to five rings depending on the respective boiling points of the individual aromatic substances. Major constituents are three- and four-ring aromatics from acenaphthene to pyrene.

3. ANALOGUE APPROACH JUSTIFICATION
Under environmental conditions or during processing of the target substance, environmentally available or volatile/water soluble components of anthracene oil can be released. These will be three- and four-ring PAH (see above). They will be distributed in the environment and may also undergo bioaccumulation in aquatic species. In combination, they will characterise the bioaccumulation potential of anthracene oil. Therefore, it is justified to use date determined for individual PAH that are constituents of anthracene oil to characterise the environmental behaviour (aquatic bioaccumulation) of anthracene oil itself.
Reason / purpose for cross-reference:
read-across source
Principles of method if other than guideline:
Read-across to preceding entry:
Source test material: polycyclic aromatic hydrocarbons (PAH), generic mixture;
Reference: Joint WHO/CONVENTION TASK FORCE 2003
Key result
Remarks on result:
other: No bioaccumulating potential of PAH in fish and mammals, and hence in the food chain, due to effective metabolism and excretion in these species
Remarks:
The test result of the source substances are adopted as weight of evidence for the target substance anthracene oil

Bioaccumulation

Aquatic organisms that metabolise PAHs to little or no extent, such as algae, molluscs and the more primitive invertebrates (protozoans, porifers and cnidaria) accumulate high concentrations of PAHs, as would be expected from their log Kow values, whereas organisms that metabolise PAHs to a great extent, such as fish and higher invertebrates, accumulate little or no PAHs.

The concentration of PAHs in vegetation is generally considerably lower than that in soil, the bioaccumulation factors ranging from 0.0001 to 0.33 for BaP and from 0.001 to 0.18 for 17 other PAHs tested including PAH constituents of anthracene oil.

Biomagnification (the increase in concentration of a substance in animals in successive trophic levels of food chains) of PAHs has not been observed in aquatic systems and would not be expected to occur, because most organisms have a high biotransformation potential for PAHs. Organisms at higher trophic levels in food chains show the highest potential for biotransformation (WHO 1998).

[from Joint WHO 2003, p.147]

Description of key information

Bioconcentration data for anthracene oil itself (benzo[a]pyrene < 50 ppm, AOL) have not been identified. But data for constituents of AOL are available. For two of them (phenanthrene and pyrene), BCF values below 2000 have been determined. The higher value of phenanthrene is used to characterise the aquatic bioconcentration of AOL.

Key value for chemical safety assessment

BCF (aquatic species):
1 082 dimensionless

Additional information

Due to the complex composition of anthracene oil, a single BCF value cannot be determined for the substance. Relevant components (all PAH) will have their individual BCF values. Phenanthrene is main constituent (ca. 25 - 31 %) and will best represent the bioaccumulation properties of anthracene oil. Hence, this substance is selected as marker substance (see Chapter 4.). In addition BCF values for pyrene, a more lipophilic constituent of anthracene oil is taken into account.

In a well-performed bioconcentration study similar/according to OECD 305 (flow-through), BCF values reported for phenanthrene ranged from 700 to 2229, depending on the exposure level (low or high) and the calculation method applied (kinetic method or steady state). At the low exposure level (0.12 µg/L; BCF values 810 and 700, respectively), less test substance accumulated in the tissue over time than at the high exposure level (1.12 µg/L; BCF values 2229 and 1623, respectively), while the very high excretion/depuration rates were the same at either exposure condition.

Lower BCF values resulted when calculating the BCFs using the ratios of tissue vs. water concentration at steady state rather than the ratio of uptake vs. elimination. Thus, the high-level BCF (kinetic BCF = 2229) fell below 2000 when based on the steady state method (steady state BCF = 1623).

For many organic substances, there is a clear relationship between the potential for bioconcentration and hydrophobicity as well as a corresponding relationship between the lipid content of the test fish and the observed bioconcentration of such substances. Thus, to reduce this source of variability in test results for those substances with high lipophilicity (i.e. with log Kow > 3), bioconcentration should be expressed as normalised to a fish with a 5 % lipid content (based on whole body wet weight) in addition to that derived directly from the study. This is necessary to provide a basis, from which results for different substances and/or test species can be compared with one another (OECD TG 305, revision of 2012).

The BCF values reported above are related to a fish lipid content of 10.3 % (ratio of lipid weight based BCF and wet weight based BCF reported in the study). Normalised to a lipid content of 5 %, the kinetic BCF values, which were higher than steady state values, decreased to 393 and 1082 for the low and high exposure concentration, respectively, thus remaining far below the cut-off value of 2000 for bioaccumulation according to Annex XIII of REACH regulation.

In the same study, pyrene was included as test substance. For this compound even lower BCF values were determined. Kinetic BCF values were higher than steady state values, as was the case for phenanthrene. Lipid-normalised kinetic BCF values were 70 and 47 for the low and high exposure concentration, respectively.

The results of the study show that the extent of bioconcentration is mainly determined by the metabolic and excretory capacity of the test organism. The high depuration rates demonstrate that the metabolic and excretory capacity was very high for both PAH in the test fish.

This study provides evidence that anthracene oil has a low to moderate bioconcentration potential based on BCF values determined for AOL constituents. However, the extent of bioconcentration is mainly determined by the metabolic and excretory capacity of the target organism.

The relevance of PAH metabolism in aquatic organisms with regard to the moderate bioaccumulation potential of PAH is also supported by data presented in a WHO report (Joint WHO/CONVENTION TASK FORCE 2003).