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Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1996-04-23 to 1996-05-4
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with different amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 19 hrs, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10l of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.4 °C
pH:
8.07 - 8.16
Dissolved oxygen:
99 - 100 % saturation
Salinity:
53 - 54 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 131, 232, 470, 886, 1734, 3379, and 5768 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.9
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
171 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 53
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
1 133 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 366
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
10 151 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 5085
Details on results:
- Mortality of test animals at end of exposure period: from 4.5 (131 mg/kg) to 81.8 % (3379 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.95)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.4°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 1133 +/- 366 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 131 to 5768 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 3 hrs in a turret mixer, and the allowed to settle for 22 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 52, 105, 233, 413, 862, 1690 and 3297 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.98 -8.05. The temperature was 15 +/- 0.6 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

8.10

100

8.09

100

131 mg/kg

8.12

100

8.10

100

232 mg/kg

8.13

100

8.11

100

470 mg/kg

8.13

100

8.13

100

886 mg/kg

8.13

100

8.15

100

1734 mg/kg

8.13

100

8.16

99

3379 mg/kg

8.13

100

8.16

100

5768 mg/kg

8.13

100

8.15

99

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

131

11

11

22

0

1

1

232

11

11

22

0

2

2

470

11

11

22

5

5

10

886

11

11

22

4

5

9

1734

11

11

22

6

6

12

3379

11

11

22

7

9

16

5768

11

11

22

9

9

18

The results mean that the 10-d LC50 is 1133 +/- 366 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

171

1133

10151

 95% Confidence interval  +/- 53  +/- 366  +/- 5085
Endpoint:
sediment toxicity: short-term
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
supporting study
Study period:
from 1996-04-23 to 1996-05-4
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with different amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 19 hrs, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10l of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.4 °C
pH:
8.07 - 8.16
Dissolved oxygen:
99 - 100 % saturation
Salinity:
53 - 54 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 131, 232, 470, 886, 1734, 3379, and 5768 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.9
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
171 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 53
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
1 133 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 366
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
10 151 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 5085
Details on results:
- Mortality of test animals at end of exposure period: from 4.5 (131 mg/kg) to 81.8 % (3379 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.95)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.4°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 1133 +/- 366 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 131 to 5768 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 3 hrs in a turret mixer, and the allowed to settle for 22 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 52, 105, 233, 413, 862, 1690 and 3297 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.98 -8.05. The temperature was 15 +/- 0.6 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

8.10

100

8.09

100

131 mg/kg

8.12

100

8.10

100

232 mg/kg

8.13

100

8.11

100

470 mg/kg

8.13

100

8.13

100

886 mg/kg

8.13

100

8.15

100

1734 mg/kg

8.13

100

8.16

99

3379 mg/kg

8.13

100

8.16

100

5768 mg/kg

8.13

100

8.15

99

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

131

11

11

22

0

1

1

232

11

11

22

0

2

2

470

11

11

22

5

5

10

886

11

11

22

4

5

9

1734

11

11

22

6

6

12

3379

11

11

22

7

9

16

5768

11

11

22

9

9

18

The results mean that the 10-d LC50 is 1133 +/- 366 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

171

1133

10151

 95% Confidence interval  +/- 53  +/- 366  +/- 5085
Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1996-06-12 to 1996-06-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 15 hrs, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10l of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.5 °C
pH:
8.01 - 8.14
Dissolved oxygen:
96 - 100 % saturation
Salinity:
54 - 56 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 109, 229, 464, 941, 1986, 3585, and 7126 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
133 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 64
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
1 211 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: 235
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
5 250 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 1124
Details on results:
- Mortality of test animals at end of exposure period: from 18.2 (109 mg/kg) to 100 % (7126 mg/kg)
- Mortality in control: 5.4 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the response and X is the exposure concentration.
(R² = 0.96)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 5.4%;

- the temperature during the exposure was 15 +/- 0.5°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 1211 +/- 460 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 109 to 7126 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 109, 229, 464, 941, 1986, 3585, and 7126 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 8.01 - 8.18. The temperature was 15 +/- 0.5 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

8.03

100

8.12

100

109 mg/kg

8.03

100

8.13

100

229 mg/kg

8.04

100

8.13

100

464 mg/kg

8.05

100

8.14

100

941 mg/kg

8.05

100

8.14

99

1986 mg/kg

8.04

100

8.13

99

3585 mg/kg

8.02

100

8.13

98

7126 mg/kg

8.03

100

8.12

97

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

1

0

1

109

11

11

22

2

2

4

229

11

11

22

2

1

3

464

11

11

22

3

5

8

941

11

11

22

4

5

9

1986

11

11

22

8

7

15

3585

11

11

22

9

9

18

7126

11

11

22

11

11

22

The results mean that the 10 -d LC50 is 1211 +/- 64 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

133

1211

5250

 95% Confidence interval  +/- 64  +/- 460  +/- 1124
Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 1996-06-12 to 1996-06-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 15 hrs, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10l of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.5 °C
pH:
8.01 - 8.14
Dissolved oxygen:
96 - 100 % saturation
Salinity:
54 - 56 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 109, 229, 464, 941, 1986, 3585, and 7126 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
133 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 64
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
1 211 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: 235
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
5 250 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 1124
Details on results:
- Mortality of test animals at end of exposure period: from 18.2 (109 mg/kg) to 100 % (7126 mg/kg)
- Mortality in control: 5.4 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the response and X is the exposure concentration.
(R² = 0.96)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 5.4%;

- the temperature during the exposure was 15 +/- 0.5°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 1211 +/- 460 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 109 to 7126 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 109, 229, 464, 941, 1986, 3585, and 7126 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 8.01 - 8.18. The temperature was 15 +/- 0.5 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

8.03

100

8.12

100

109 mg/kg

8.03

100

8.13

100

229 mg/kg

8.04

100

8.13

100

464 mg/kg

8.05

100

8.14

100

941 mg/kg

8.05

100

8.14

99

1986 mg/kg

8.04

100

8.13

99

3585 mg/kg

8.02

100

8.13

98

7126 mg/kg

8.03

100

8.12

97

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

1

0

1

109

11

11

22

2

2

4

229

11

11

22

2

1

3

464

11

11

22

3

5

8

941

11

11

22

4

5

9

1986

11

11

22

8

7

15

3585

11

11

22

9

9

18

7126

11

11

22

11

11

22

The results mean that the 10 -d LC50 is 1211 +/- 64 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

133

1211

5250

 95% Confidence interval  +/- 64  +/- 460  +/- 1124
Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1996-03-17 to 1996-05-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 16 hors, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10L of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.4 °C
pH:
8.10 - 8.18
Dissolved oxygen:
99 - 100 % saturation
Salinity:
53 - 54 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 128, 206, 376, 761, 1530, 3012, and 5497 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 ml glass beakers (tall form)
- Volume of sediment: 200 ml
- Overlying water volume: 650 ml

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.9
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
174 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 22
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
827 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 114
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 5 497 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Details on results:
- Mortality of test animals at end of exposure period: from 0 (128 mg/kg) to 86.4 % (5497 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 1.00)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.4°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 827 +/- 114 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 128 to 5497 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 128, 206, 376, 761, 1530, 3012, and 5497 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 8.10 -8.18. The temperature was 15 +/- 0.4 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

8.13

100

8.16

100

128 mg/kg

8.14

100

8.16

100

206 mg/kg

8.13

100

8.16

100

376 mg/kg

8.14

100

817

100

761 mg/kg

8.14

100

8.17

99

1530 mg/kg

8.14

100

8.18

100

3012 mg/kg

8.13

100

8.17

99

5476 mg/kg

8.14

100

8.18

99

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

52

11

11

22

0

0

0

105

11

11

22

2

2

4

233

11

11

22

3

3

6

413

11

11

22

5

6

11

862

11

11

22

7

7

14

1690

11

11

22

8

9

17

3297

11

11

22

9

10

19

The results mean that the 10 -d LC50 is 827 +/- 114 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

Sample B

174

827

>5497

 95% Confidence interval  +/- 22  +/- 114  -
Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 1996-03-17 to 1996-05-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 16 hors, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10L of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.4 °C
pH:
8.10 - 8.18
Dissolved oxygen:
99 - 100 % saturation
Salinity:
53 - 54 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 128, 206, 376, 761, 1530, 3012, and 5497 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 ml glass beakers (tall form)
- Volume of sediment: 200 ml
- Overlying water volume: 650 ml

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.9
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
174 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 22
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
827 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 114
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 5 497 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Details on results:
- Mortality of test animals at end of exposure period: from 0 (128 mg/kg) to 86.4 % (5497 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 1.00)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.4°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 827 +/- 114 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 128 to 5497 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 128, 206, 376, 761, 1530, 3012, and 5497 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 8.10 -8.18. The temperature was 15 +/- 0.4 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

8.13

100

8.16

100

128 mg/kg

8.14

100

8.16

100

206 mg/kg

8.13

100

8.16

100

376 mg/kg

8.14

100

817

100

761 mg/kg

8.14

100

8.17

99

1530 mg/kg

8.14

100

8.18

100

3012 mg/kg

8.13

100

8.17

99

5476 mg/kg

8.14

100

8.18

99

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

52

11

11

22

0

0

0

105

11

11

22

2

2

4

233

11

11

22

3

3

6

413

11

11

22

5

6

11

862

11

11

22

7

7

14

1690

11

11

22

8

9

17

3297

11

11

22

9

10

19

The results mean that the 10 -d LC50 is 827 +/- 114 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

Sample B

174

827

>5497

 95% Confidence interval  +/- 22  +/- 114  -
Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1998-10-19 to 1998-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 16 hors, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10L of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 1.5 °C
pH:
7.98 - 8.05
Dissolved oxygen:
93 - 97 % saturation
Salinity:
46.1 - 47.2 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 52, 105, 233, 413, 862, 1690, and 3297 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
91 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: Conf. Interv. 52-130
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
680 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: Conf. Interv. 349- - 1011
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
>= 3 297 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Details on results:
- Mortality of test animals at end of exposure period: from 9.1 (52 mg/kg) to 72.7 % (3297 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.83)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.6°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 680 +/- 331 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 52 to 3297 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 52, 105, 233, 413, 862, 1690 and 3297 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.98 -8.05. The temperature was 15 +/- 0.6 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

7..99

96

8.02

96

52 mg/kg

7.99

95

8.02

96

105 mg/kg

7.99

97

8.02

94

233 mg/kg

8.01

96

8.04

93

413 mg/kg

8.01

97

8.05

93

862 mg/kg

8.01

97

8.04

94

1690 mg/kg

8.03

96

8.05

94

3297 mg/kg

8.02

97

8.06

95

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

Only slight deviations from the protocol were noted noted (adjustment of salinity at day 5; no observations of oxygen saturation, salinity nor pH at day 5). No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

52

11

11

22

1

1

2

105

11

11

22

0

1

1

233

11

11

22

0

2

2

413

11

11

22

5

7

12

862

11

11

22

6

8

14

1690

11

11

22

6

7

13

3297

11

11

22

8

8

16

The results mean that the 10 -d LC50 is 680 +/- 331 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

91

680

>3297

 95% Confidence interval  +/- 39    
Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 1998-10-19 to 1998-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 16 hors, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10L of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 1.5 °C
pH:
7.98 - 8.05
Dissolved oxygen:
93 - 97 % saturation
Salinity:
46.1 - 47.2 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 52, 105, 233, 413, 862, 1690, and 3297 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
91 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: Conf. Interv. 52-130
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
680 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: Conf. Interv. 349- - 1011
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
>= 3 297 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Details on results:
- Mortality of test animals at end of exposure period: from 9.1 (52 mg/kg) to 72.7 % (3297 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.83)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.6°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 680 +/- 331 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 52 to 3297 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 52, 105, 233, 413, 862, 1690 and 3297 mg/kg. Two replicateswith at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.98 -8.05. The temperature was 15 +/- 0.6 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

7..99

96

8.02

96

52 mg/kg

7.99

95

8.02

96

105 mg/kg

7.99

97

8.02

94

233 mg/kg

8.01

96

8.04

93

413 mg/kg

8.01

97

8.05

93

862 mg/kg

8.01

97

8.04

94

1690 mg/kg

8.03

96

8.05

94

3297 mg/kg

8.02

97

8.06

95

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

Only slight deviations from the protocol were noted noted (adjustment of salinity at day 5; no observations of oxygen saturation, salinity nor pH at day 5). No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

52

11

11

22

1

1

2

105

11

11

22

0

1

1

233

11

11

22

0

2

2

413

11

11

22

5

7

12

862

11

11

22

6

8

14

1690

11

11

22

6

7

13

3297

11

11

22

8

8

16

The results mean that the 10 -d LC50 is 680 +/- 331 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

91

680

>3297

 95% Confidence interval  +/- 39    
Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1998-12-09 to 1998-12-20
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 16 hours, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10L of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.8 °C
pH:
7.96 - 8.05
Dissolved oxygen:
93 - 97 % saturation
Salinity:
46.9 - 47.3 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 84, 160, 336, 634, 1229, 2409, and 4649 mg/kg sediment dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
99 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 26
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
312 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 87
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
1 244 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 531
Details on results:
- Mortality of test animals at end of exposure period: from 9.1 (84 mg/kg) to 100 % (4649 mg/kg)
- Mortality in control: 4.5 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.96)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.6°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 312 +/- 87 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 84 to 4649 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 84, 160, 3336, 634, 1229, 2409, and 4649 mg/kg. Two replicates with at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.96 -8.05. The temperature was 15 +/- 0.8 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

7..99

96

8.02

96

84 mg/kg

7.99

95

8.02

96

160 mg/kg

8.01

96

8.04

93

336 mg/kg

8.01

97

8.05

93

634 mg/kg

8.01

97

8.04

94

1229 mg/kg

8.03

96

8.05

94

4649 mg/kg

8.02

97

8.06

95

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

1

0

2

84

11

11

22

1

1

2

160

11

11

22

2

3

5

336

11

11

22

6

7

13

634

11

11

22

10

9

19

1229

11

11

22

9

8

17

2409

11

11

22

6

10

16

464997

11

11

22

11

11

22

The results mean that the 10 -d LC50 is 312 +/- 87 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -31

99

312

1244

 95% Confidence interval  +/- 26  +/- 87  +/- 531
Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 1998-12-09 to 1998-12-20
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well documented guideline study.
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site (Mid Norway) as the amphipods (organic content between 0.5 an 4%; silt/clay fraction (<63µm) between 5 and 20%; median grain size of 90 to 125 µm), collected from an intertidal mud flats at eb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polethylene trays with seawater poured on top of it, transported to the laboratory, kept at 8 +/- 2°C and sieved within 48 hrs to 500 µm in reference seawater (natural seawater with a salinity of 32-34 per thousand, filtered to <70-80 µm by gravel filter and aerated to saturation before use). Before use, the sediment is resuspended in reference seawater and thouroughly homogenised.

- Spiked test sediment: Reference sediment is spiked with differnet amounts of test substance in 1-L HD polyethylene bottle in a turret mixer at 55-60 rev/min for 3 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 16 hours, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population in mid Norway considered free from contaminants. Corophium are collected from clean intertidal mud flats at ebb tide by gently removing the upper 5 cm of the sediment with a spade and transferring it to HD polyethylene trays. Seawater is poured on top of the sediment, and the trays are transported to the laboratory, placed at 8 +/- 2 °C and sieved within 48 hrs to 500µM with full salinity seawater. The fraction collected in the sieve is transferred to an aquarium with seawater.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long

ACCLIMATION
- Acclimation period: 3 days
- Acclimation conditions (same as test or not): temperature 15+/- 1.5 °C; in aquaria with aeration and 5-10L of reference seawater
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.8 °C
pH:
7.96 - 8.05
Dissolved oxygen:
93 - 97 % saturation
Salinity:
46.9 - 47.3 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 84, 160, 336, 634, 1229, 2409, and 4649 mg/kg sediment dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)
- Volume of sediment: 200 mL
- Overlying water volume: 650 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


NATURAL SEDIMENT CHARACTERISTICS
- Natural marine sediment from the habitat of the amphipods free of known contaminants, collected from the 5 cm upper layer of sediment of intertidal mud flats at ebb tide (Mid Norway).
- Orgnaic content between 0.5 and 4 %
- silt/clay fraction (<63 µm) between 5 and 20%
- median grain size : 90 - 125 µm


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
99 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 26
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
312 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 87
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
1 244 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 531
Details on results:
- Mortality of test animals at end of exposure period: from 9.1 (84 mg/kg) to 100 % (4649 mg/kg)
- Mortality in control: 4.5 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.96)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.6°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 312 +/- 87 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 84 to 4649 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >70 -80 µm by gravel filters and aerated to saturation before use. The test substance was directly added to the sediment and the spiked sediment was mixed for 2 hrs in a turret mixer, and the allowed to settle for 16 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 84, 160, 3336, 634, 1229, 2409, and 4649 mg/kg. Two replicates with at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.96 -8.05. The temperature was 15 +/- 0.8 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

7..99

96

8.02

96

84 mg/kg

7.99

95

8.02

96

160 mg/kg

8.01

96

8.04

93

336 mg/kg

8.01

97

8.05

93

634 mg/kg

8.01

97

8.04

94

1229 mg/kg

8.03

96

8.05

94

4649 mg/kg

8.02

97

8.06

95

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

1

0

2

84

11

11

22

1

1

2

160

11

11

22

2

3

5

336

11

11

22

6

7

13

634

11

11

22

10

9

19

1229

11

11

22

9

8

17

2409

11

11

22

6

10

16

464997

11

11

22

11

11

22

The results mean that the 10 -d LC50 is 312 +/- 87 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -31

99

312

1244

 95% Confidence interval  +/- 26  +/- 87  +/- 531
Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1999-06-25 to 1999-07-05
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods, sieved to remove organisms and debris.

- Spiked test sediment: calculated amoounts of test substance were aded to the vessels containing the wet sediment and mixed thouroughly with a clean spatula. 100 mL of artificial sea water (Coral Reef, Ken Sea Salt, Red Sea Fish Pharm) with a salinity between 25 and 35 per thousand were then added to each vessel.

- The sediment in the controls was treated like the test sediment but without adding test substance.

Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
Batch freshly dug from the North Wales coast, free of malformations and of good health obtained on 1999-06-18 (batch number: CV990006)

ACCLIMATION
Acclimation conditions: stock amphipods were held in 13-l storage vessels containing sieved sediment obtained from the collection site.
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15.0 - 16.9 °C
pH:
7.88 - 8.21
Dissolved oxygen:
94 -99 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 100.0, 177.8, 316.2, 562.3 and 1000.0 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): Tall form glass beakers, 1-L capacity
- Volume of wet sediment: no data
- Overlying water volume: 100 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 20
- No. of replicates per treatment group: 3
- No. of replicates per control: 5


OVERLYING WATER CHARACTERISTCS
- Artificial seawater (Coral Reef, Kent Sea Salt, Red Sea Fish Pharm
- Salinity: 25 - 35 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality

Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
299.6 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Confid. Interv. 200.5 - 441.5
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
100 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 7 (100 mg/kg dw) to 90 % (1000.0 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
LC50 value calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board

No remarks

Validity criteria fulfilled:
not specified
Conclusions:
10-d LC50 = 299.6 mg/kg sediment dw
10-d NOEC = 100.0 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 100.0 to 1000.0 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was artificial seawater (Coral reef, Kent Sea Salt, Red Sea Fish Pharm). The test substance was directly added to the sediment and mixed thouroughly with a clean spatula. 100 mL of artificial seawater was added to each 1 -L beaker. The controls were prepared the same way. In the test, 5 concentrations were tested: 100.0, 177.8, 316.2, 562.3, and 1000.0 1495 mg/kg. Three replicates were used for each concentration and five for the control.

Dissolved oxygen was >94% during the study. The pH was 7.88 - 8.21. The temperature was 15.0 - 16.9 °C.

Test substance concentration

Oxygen (%)

pH

Mean

Std Dev.

Mean

Std Dev.

Control

95.9

1.2

8.04

0.07

100.0 mg/kg dw

96.0

1.2

8.11

0.07

177.8 mg/kg dw

95.8

1.2

8.11

0.06

316.2 mg/kg dw

95.9

1.2

8.11

0.06

569.3 mg/kg dw

95.9

1.3

8.12

0.06

1000.0 mg/kg dw

95.8

0.8

8.09

0.06

(Means of measurements in all replicates on day 0, 1, 4, 7, and 10)

The test organisms were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Tank n°

Mortality

Day 1

Day 4

Day 7

Day 10

End

Total (%)

Control

A

0

0

0

0

0

5

B

0

0

0

0

4

C

0

0

0

0

1

D

0

0

0

0

0

E

0

0

0

0

0

100.0

A

0

0

0

0

0

7

B

0

0

0

0

1

C

0

0

0

0

3

177.8

A

0

0

0

0

3

32

B

0

0

0

0

6

C

0

0

0

0

10

316.2

A

0

2

0

0

7

43

B

0

0

0

0

12

C

0

1

0

0

7

562.3

A

0

1

0

0

18

87

B

0

0

2

17

16

C

0

0

1

8

18

1000.0

A

0

0

1

0

16

90

B

0

0

0

0

19

C

0

1

0

0

19

The results mean that the 10-d LC50 is 299.6 mg/kg sediment dry weight and the 10-d NOEC is 100.0 mg/kg sediment dry weight.

Endpoint:
sediment toxicity: short-term
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
supporting study
Study period:
from 1999-06-25 to 1999-07-05
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods, sieved to remove organisms and debris.

- Spiked test sediment: calculated amoounts of test substance were aded to the vessels containing the wet sediment and mixed thouroughly with a clean spatula. 100 mL of artificial sea water (Coral Reef, Ken Sea Salt, Red Sea Fish Pharm) with a salinity between 25 and 35 per thousand were then added to each vessel.

- The sediment in the controls was treated like the test sediment but without adding test substance.

Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
Batch freshly dug from the North Wales coast, free of malformations and of good health obtained on 1999-06-18 (batch number: CV990006)

ACCLIMATION
Acclimation conditions: stock amphipods were held in 13-l storage vessels containing sieved sediment obtained from the collection site.
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15.0 - 16.9 °C
pH:
7.88 - 8.21
Dissolved oxygen:
94 -99 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 100.0, 177.8, 316.2, 562.3 and 1000.0 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): Tall form glass beakers, 1-L capacity
- Volume of wet sediment: no data
- Overlying water volume: 100 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 20
- No. of replicates per treatment group: 3
- No. of replicates per control: 5


OVERLYING WATER CHARACTERISTCS
- Artificial seawater (Coral Reef, Kent Sea Salt, Red Sea Fish Pharm
- Salinity: 25 - 35 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality

Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
299.6 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Confid. Interv. 200.5 - 441.5
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
100 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 7 (100 mg/kg dw) to 90 % (1000.0 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
LC50 value calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board

No remarks

Validity criteria fulfilled:
not specified
Conclusions:
10-d LC50 = 299.6 mg/kg sediment dw
10-d NOEC = 100.0 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 100.0 to 1000.0 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was artificial seawater (Coral reef, Kent Sea Salt, Red Sea Fish Pharm). The test substance was directly added to the sediment and mixed thouroughly with a clean spatula. 100 mL of artificial seawater was added to each 1 -L beaker. The controls were prepared the same way. In the test, 5 concentrations were tested: 100.0, 177.8, 316.2, 562.3, and 1000.0 1495 mg/kg. Three replicates were used for each concentration and five for the control.

Dissolved oxygen was >94% during the study. The pH was 7.88 - 8.21. The temperature was 15.0 - 16.9 °C.

Test substance concentration

Oxygen (%)

pH

Mean

Std Dev.

Mean

Std Dev.

Control

95.9

1.2

8.04

0.07

100.0 mg/kg dw

96.0

1.2

8.11

0.07

177.8 mg/kg dw

95.8

1.2

8.11

0.06

316.2 mg/kg dw

95.9

1.2

8.11

0.06

569.3 mg/kg dw

95.9

1.3

8.12

0.06

1000.0 mg/kg dw

95.8

0.8

8.09

0.06

(Means of measurements in all replicates on day 0, 1, 4, 7, and 10)

The test organisms were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Tank n°

Mortality

Day 1

Day 4

Day 7

Day 10

End

Total (%)

Control

A

0

0

0

0

0

5

B

0

0

0

0

4

C

0

0

0

0

1

D

0

0

0

0

0

E

0

0

0

0

0

100.0

A

0

0

0

0

0

7

B

0

0

0

0

1

C

0

0

0

0

3

177.8

A

0

0

0

0

3

32

B

0

0

0

0

6

C

0

0

0

0

10

316.2

A

0

2

0

0

7

43

B

0

0

0

0

12

C

0

1

0

0

7

562.3

A

0

1

0

0

18

87

B

0

0

2

17

16

C

0

0

1

8

18

1000.0

A

0

0

1

0

16

90

B

0

0

0

0

19

C

0

1

0

0

19

The results mean that the 10-d LC50 is 299.6 mg/kg sediment dry weight and the 10-d NOEC is 100.0 mg/kg sediment dry weight.

Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1999- 05-22 to 1999-06-02
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment (collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Qualifier:
according to guideline
Guideline:
other: HOCNF (Revised Harmonised Offshore Notification Format) document from the OSPAR Commission, 1995
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site as the amphipods, washed freely through a 0.5 mm sieve with dilution seawater.

- Spiked test sediment: Reference sediment is spiked with different amounts of test substance in 1-L HD polyethylene bottle in a turret mixer for 1.5 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 20 hrs, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium sp.
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population, held static in their natural sediment in HD polyethylene trays in dilution seawater at 10 +/- 3°C. Water is renewed weekly. As they are kept in their natural sediment, the animals are not fed in the laboratory.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.7 °C
pH:
7.98 - 8.06
Dissolved oxygen:
94 - 97 % saturation
Salinity:
46.9 - 47.7 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 97, 185, 303, 588, 1061, 1918, and 3133 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
110 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 19
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
292 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 56
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
952 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 269
Details on results:
- Mortality of test animals at end of exposure period: from 13.6 (97 mg/kg) to 100 % (3133 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.95)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.6°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 292 +/- 56 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 27 to 3133 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >5 µm. The test substance was directly added to the sediment and the spiked sediment was mixed for 1.5 hrs in a turret mixer, and the allowed to settle for 22 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 97, 185, 303, 588, 1061, 1918, and 3133 mg/kg. Two replicates with at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.97 -8.05. The temperature was 15 +/- 0.7 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

7.98

96

7.99

96

97 mg/kg

7.99

96

8.02

97

185 mg/kg

7.99

96

8.01

96

303 mg/kg

7.99

96

8.03

97

588 mg/kg

7.99

96

8.04

95

1061 mg/kg

8.00

95

8.03

95

1918 mg/kg

8.00

95

8.03

96

3133 mg/kg

8.01

96

8.06

96

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

97

11

11

22

1

2

3

185

11

11

22

3

2

5

303

11

11

22

4

5

9

588

11

11

22

9

10

19

1061

11

11

22

10

11

21

1918

11

11

22

11

11

22

3133

11

11

22

11

11

22

The results mean that the 10 -d LC50 is 292 +/- 56 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

110

292

952

 95% Confidence interval  +/- 19  +/- 56  +/- 269
Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 1999- 05-22 to 1999-06-02
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment (collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Ring test Protocol: A sediment bioassay using an amphipod Corophium sp. (McMinn, W., Thain, J., van den Hurk, P., & Roddie, B.)
Deviations:
yes
Remarks:
Adjustment of salinity in exposure vessels at day 5; no observations of pH, oxygen saturation or salinity at day 5.
Qualifier:
according to guideline
Guideline:
other: HOCNF (Revised Harmonised Offshore Notification Format) document from the OSPAR Commission, 1995
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Reference sediment: Natural sediment from the same unpolluted site as the amphipods, washed freely through a 0.5 mm sieve with dilution seawater.

- Spiked test sediment: Reference sediment is spiked with different amounts of test substance in 1-L HD polyethylene bottle in a turret mixer for 1.5 hrs in an axial manner. The test substance is mixed with dried sediment and added directly to the mixing bottles without the use of any solvent. After mixing, the spiked sediment is transferred to 1-L beakers (tall form). After sedimentation for 20 hrs, a LD polyethylene sheet was placed on top of the sediment and aerated refence seawater was added before transferrel of the amphipods.

- The sediment in the controls was treated like the test sediment but without adding test substance.
Test organisms (species):
Corophium sp.
Details on test organisms:
TEST ORGANISM
- Details on collection: animals collected from a natural population, held static in their natural sediment in HD polyethylene trays in dilution seawater at 10 +/- 3°C. Water is renewed weekly. As they are kept in their natural sediment, the animals are not fed in the laboratory.
- Age and size of animals at beginning of exposure: Adult stage >5 mm long
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 0.7 °C
pH:
7.98 - 8.06
Dissolved oxygen:
94 - 97 % saturation
Salinity:
46.9 - 47.7 mS
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 97, 185, 303, 588, 1061, 1918, and 3133 mg/kg sediment

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 1000 mL glass beakers (tall form)

EXPOSURE REGIME
- No. of organisms per container (treatment): at least 11
- No. of replicates per treatment group: 2
- No. of replicates per control: 2


OVERLYING WATER CHARACTERISTCS
Natural sea water with a salinity of 32-34 per thousand filtered to <70-80 µm by gravel filters and aerated to saturation before use.


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Immobilisation
Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
110 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 19
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
292 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 56
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
952 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
behaviour
Remarks:
immobilisation
Remarks on result:
other: +/- 269
Details on results:
- Mortality of test animals at end of exposure period: from 13.6 (97 mg/kg) to 100 % (3133 mg/kg)
- Mortality in control: 0 %
Results with reference substance (positive control):
Not applicable (no reference substance)
Reported statistics and error estimates:
Calculation of the LC-values was based on the non-linear model: Y = aX + blogX + c where Y is the resposne and X is the exposure concentration.
(R² = 0.95)

Test acceptability:

The test acceptance criteria given were fulfilled :

- the immobilisation in the control was 0%;

- the temperature during the exposure was 15 +/- 0.6°C;

- saturation of oxygen was at least 93 %.

Validity criteria fulfilled:
yes
Remarks:
Temperature 15 +/- 0.6 °C; 0% immobilisation in the control; saturation of oxygen>= 93%
Conclusions:
10-d LC50 = 292 +/- 56 mg/kg sediment
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 27 to 3133 mg/kg sediment. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site with a salinity 0f 32 -34 per thousand filtered to >5 µm. The test substance was directly added to the sediment and the spiked sediment was mixed for 1.5 hrs in a turret mixer, and the allowed to settle for 22 hrs. The controls were prepared the same way. In the test, 7 concentrations were tested in a geometric series: 97, 185, 303, 588, 1061, 1918, and 3133 mg/kg. Two replicates with at least 11 amphipods each, were used for each concentration and the control.

Dissolved oxygen was >93% saturation during the study. The pH was 7.97 -8.05. The temperature was 15 +/- 0.7 °C.

Test substance concentration

Initial values

Day 10

pH

O2%

pH

O2%

Control

7.98

96

7.99

96

97 mg/kg

7.99

96

8.02

97

185 mg/kg

7.99

96

8.01

96

303 mg/kg

7.99

96

8.03

97

588 mg/kg

7.99

96

8.04

95

1061 mg/kg

8.00

95

8.03

95

1918 mg/kg

8.00

95

8.03

96

3133 mg/kg

8.01

96

8.06

96

(The values are mean values of replicates)

The test organisms (adults >5 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Nominal concentration (mg/kg)

Number of Corophium volutator

Initial

Immobilised after 10 days

Replicate 1

Replicate2

Total

Replicate 1

Replicate2

Total

Control

11

11

22

0

0

0

97

11

11

22

1

2

3

185

11

11

22

3

2

5

303

11

11

22

4

5

9

588

11

11

22

9

10

19

1061

11

11

22

10

11

21

1918

11

11

22

11

11

22

3133

11

11

22

11

11

22

The results mean that the 10 -d LC50 is 292 +/- 56 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

DEV 98 -25

110

292

952

 95% Confidence interval  +/- 19  +/- 56  +/- 269
Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 1999-02-08 to 1999-02-18
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment (collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods, sieved to remove organisms and debris.

- Spiked test sediment: calculated amoounts of test substance were aded to the vessels containing the wet sediment and mixed thouroughly with a clean spatula. 100 mL of artificial sea water (Coral Reef, Ken Sea Salt, Red Sea Fish Pharm) with a salinity between 25 and 35 per thousand were then added to each vessel.

- The sediment in the controls was treated like the test sediment but without adding test substance.

Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
Batch freshly dug from the North Wales coast, free of malformations and of good health supplied by MBCC on 1999-02-05

ACCLIMATION
Acclimation conditions (same as test or not): stock amphipods were held in 13-L storage vessels containing sieved sediment obtained from the collection site.
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
13- 15.5 °C
pH:
8.16 - 8.36
Dissolved oxygen:
89 - 100 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 56.2, 100.0, 177.8, 316.2, 562.3 and 1000.0 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): Tall form glass beakers, 1-L capacity
- Volume of wet sediment: no data
- Overlying water volume: 100 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 20
- No. of replicates per treatment group: 3
- No. of replicates per control: 5


OVERLYING WATER CHARACTERISTCS
- Artificial seawater (Coral Reef, Kent Sea Salt, Red Sea Fish Pharm
- Salinity: 25 - 35 per thousand

SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality

Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
200.3 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Confid. Interv. 100.2 - 355.3
Details on results:
- Mortality of test animals at end of exposure period: from 28 (56.2 mg/kg dw) to 88 % (1000.0 mg/kg dw)
- Mortality in control: 7 %
Results with reference substance (positive control):
- Relevant effect levels: LC50 = 200.3 mg/kg dw (95% confidence interval: 100.2 - 355.3 mg/kg dw)
Reported statistics and error estimates:
LC50 value calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board

No remarks

Validity criteria fulfilled:
not specified
Conclusions:
10-d LC50 = 200.3 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 56.2 to 1000.0 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was artificial seawater (Coral reef, Kent Sea Salt, Red Sea Fish Pharm). The test substance was directly added to the sediment and mixed thouroughly with a clean spatula. 100 mL of artificial seawater was added to each 1 -L beaker. The controls were prepared the same way. In the test, 6 concentrations were tested: 56.2, 100.0, 177.8, 316.2, 562.3, and 1000.0 1495 mg/kg. Three replicates were used for each concentration and five for the control.

Dissolved oxygen was >89% during the study. The pH was 8.16 -8.36. The temperature was 13 - 15.5 °C.

Test substance concentration

Oxygen (%)

pH

Mean

Std Dev.

Mean

Std Dev.

Control

96

2.5

8.23

0.05

56.2 mg/kg dw

95

2.9

8.29

0.06

100.0 mg/kg dw

94

2.9

8.29

0.05

177.8 mg/kg dw

95

2.7

8.29

0.05

316.2 mg/kg dw

95

2.4

8.30

0.06

569.3 mg/kg dw

95

3.3

8.31

0.06

1000.0 mg/kg dw

95

3.5

8.31

0.05

(Means of measurements in all replicates at day 0, 1, 4, 7, and 10)

The test organisms were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Tank n°

Mortality

Day 1

Day 4

Day 7

Day 10

End

Total (%)

Control

A

0

0

0

0

0

7

B

0

0

0

0

3

C

0

0

0

0

1

D

0

0

0

0

2

E

0

0

0

0

1

56.2

A

0

0

0

0

8

28

B

0

0

1

0

2

C

0

0

0

0

7

100.0

A

0

0

0

0

7

43

B

0

0

2

1

9

C

0

0

0

0

10

177.8

A

0

0

0

1

9

35

B

0

0

0

0

5

C

0

0

0

0

7

316.2

A

0

0

0

0

9

57

B

0

0

0

0

13

C

0

0

0

0

12

562.3

A

0

0

0

0

12

67

B

0

0

0

0

14

C

0

0

0

0

14

1000.0

A

0

0

1

0

19

88

B

0

0

0

0

18

C

0

0

0

0

16

The results mean that the 10 -d LC50 is 200.3 mg/kg sediment dry weight.

Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 1999-02-08 to 1999-02-18
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment (collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
no
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods, sieved to remove organisms and debris.

- Spiked test sediment: calculated amoounts of test substance were aded to the vessels containing the wet sediment and mixed thouroughly with a clean spatula. 100 mL of artificial sea water (Coral Reef, Ken Sea Salt, Red Sea Fish Pharm) with a salinity between 25 and 35 per thousand were then added to each vessel.

- The sediment in the controls was treated like the test sediment but without adding test substance.

Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
Batch freshly dug from the North Wales coast, free of malformations and of good health supplied by MBCC on 1999-02-05

ACCLIMATION
Acclimation conditions (same as test or not): stock amphipods were held in 13-L storage vessels containing sieved sediment obtained from the collection site.
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
13- 15.5 °C
pH:
8.16 - 8.36
Dissolved oxygen:
89 - 100 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 56.2, 100.0, 177.8, 316.2, 562.3 and 1000.0 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): Tall form glass beakers, 1-L capacity
- Volume of wet sediment: no data
- Overlying water volume: 100 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 20
- No. of replicates per treatment group: 3
- No. of replicates per control: 5


OVERLYING WATER CHARACTERISTCS
- Artificial seawater (Coral Reef, Kent Sea Salt, Red Sea Fish Pharm
- Salinity: 25 - 35 per thousand

SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality

Reference substance (positive control):
no
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
200.3 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Confid. Interv. 100.2 - 355.3
Details on results:
- Mortality of test animals at end of exposure period: from 28 (56.2 mg/kg dw) to 88 % (1000.0 mg/kg dw)
- Mortality in control: 7 %
Results with reference substance (positive control):
- Relevant effect levels: LC50 = 200.3 mg/kg dw (95% confidence interval: 100.2 - 355.3 mg/kg dw)
Reported statistics and error estimates:
LC50 value calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board

No remarks

Validity criteria fulfilled:
not specified
Conclusions:
10-d LC50 = 200.3 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 56.2 to 1000.0 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was artificial seawater (Coral reef, Kent Sea Salt, Red Sea Fish Pharm). The test substance was directly added to the sediment and mixed thouroughly with a clean spatula. 100 mL of artificial seawater was added to each 1 -L beaker. The controls were prepared the same way. In the test, 6 concentrations were tested: 56.2, 100.0, 177.8, 316.2, 562.3, and 1000.0 1495 mg/kg. Three replicates were used for each concentration and five for the control.

Dissolved oxygen was >89% during the study. The pH was 8.16 -8.36. The temperature was 13 - 15.5 °C.

Test substance concentration

Oxygen (%)

pH

Mean

Std Dev.

Mean

Std Dev.

Control

96

2.5

8.23

0.05

56.2 mg/kg dw

95

2.9

8.29

0.06

100.0 mg/kg dw

94

2.9

8.29

0.05

177.8 mg/kg dw

95

2.7

8.29

0.05

316.2 mg/kg dw

95

2.4

8.30

0.06

569.3 mg/kg dw

95

3.3

8.31

0.06

1000.0 mg/kg dw

95

3.5

8.31

0.05

(Means of measurements in all replicates at day 0, 1, 4, 7, and 10)

The test organisms were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Tank n°

Mortality

Day 1

Day 4

Day 7

Day 10

End

Total (%)

Control

A

0

0

0

0

0

7

B

0

0

0

0

3

C

0

0

0

0

1

D

0

0

0

0

2

E

0

0

0

0

1

56.2

A

0

0

0

0

8

28

B

0

0

1

0

2

C

0

0

0

0

7

100.0

A

0

0

0

0

7

43

B

0

0

2

1

9

C

0

0

0

0

10

177.8

A

0

0

0

1

9

35

B

0

0

0

0

5

C

0

0

0

0

7

316.2

A

0

0

0

0

9

57

B

0

0

0

0

13

C

0

0

0

0

12

562.3

A

0

0

0

0

12

67

B

0

0

0

0

14

C

0

0

0

0

14

1000.0

A

0

0

1

0

19

88

B

0

0

0

0

18

C

0

0

0

0

16

The results mean that the 10 -d LC50 is 200.3 mg/kg sediment dry weight.

Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2006-10-03 to 2006-10-14
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 Ll acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 23.09.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days (29.09-0.3.10.2006)
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.1
Dissolved oxygen:
88-96 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 126, 308, 668, 1495 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


HANDLING OF NATURAL SEDIMENT
- Time of collection: 23.09.2006


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.28
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Key result
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
207 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
503 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
1 220 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
126 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 3.7 (126 mg/kg dw) to 96.3 % (96.3 mg/kg dw)
- Mortality in control: 3.3 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 13.3 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 13.3 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 503 mg/kg sediment dw
10-d NOEC = 126 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 126 to 1495 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 126, 308, 668 and 1495 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40% during the study. The pH was 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.1

95

96

96

126 mg/kg dw

8.1

95

96

90

308 mg/kg dw

8.1

95

96

93

668 mg/kg dw

8.1

95

93

88

1495 mg/kg dw

8.1

96

90

93

Reference, 5.57 mg/kg dw

8.1

95

90

96

Reference, 10.3 mg/kg dw

8.1

95

90

93

Reference, 18.9 mg/kg dw

8.1

95

93

95

Reference, 32.0 mg/kg dw

8.1

95

96

91

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

2

0

0

0

0

0

2

126

27

9

9

9

1

0

0

1

308

27

9

9

9

3

1

2

6

668

27

9

9

9

6

5

6

17

1495

27

9

9

9

8

9

9

26

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 10.3

27

9

9

9

2

3

3

8

Ref., 18.9

27

9

9

9

8

7

6

21

Ref., 32.0

27

9

9

9

9

9

9

27

The results mean that the 10 -d LC50 is 503 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

8.2

13.3

21.6

5.57

DEV 2006-16

207

503

1220

126

Endpoint:
sediment toxicity: short-term
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
from 2006-10-03 to 2006-10-14
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 Ll acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 23.09.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days (29.09-0.3.10.2006)
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.1
Dissolved oxygen:
88-96 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 126, 308, 668, 1495 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


HANDLING OF NATURAL SEDIMENT
- Time of collection: 23.09.2006


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.28
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Key result
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
207 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
503 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
1 220 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
126 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 3.7 (126 mg/kg dw) to 96.3 % (96.3 mg/kg dw)
- Mortality in control: 3.3 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 13.3 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 13.3 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 503 mg/kg sediment dw
10-d NOEC = 126 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 126 to 1495 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 126, 308, 668 and 1495 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40% during the study. The pH was 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.1

95

96

96

126 mg/kg dw

8.1

95

96

90

308 mg/kg dw

8.1

95

96

93

668 mg/kg dw

8.1

95

93

88

1495 mg/kg dw

8.1

96

90

93

Reference, 5.57 mg/kg dw

8.1

95

90

96

Reference, 10.3 mg/kg dw

8.1

95

90

93

Reference, 18.9 mg/kg dw

8.1

95

93

95

Reference, 32.0 mg/kg dw

8.1

95

96

91

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

2

0

0

0

0

0

2

126

27

9

9

9

1

0

0

1

308

27

9

9

9

3

1

2

6

668

27

9

9

9

6

5

6

17

1495

27

9

9

9

8

9

9

26

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 10.3

27

9

9

9

2

3

3

8

Ref., 18.9

27

9

9

9

8

7

6

21

Ref., 32.0

27

9

9

9

9

9

9

27

The results mean that the 10 -d LC50 is 503 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

8.2

13.3

21.6

5.57

DEV 2006-16

207

503

1220

126

Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with ant without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
91-98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 130, 302, 671, and 1500 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.26
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
172 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
755 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 500 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
302 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 3.3 (130 mg/kg dw) to 70 % (1500 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 755 mg/kg sediment dw
10-d NOEC = 302 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 130 to 1500 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 130, 302, 671, and 1500 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 % during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

130 mg/kg dw

8.0

93

94

95

302 mg/kg dw

8.1

95

94

95

671 mg/kg dw

8.0

95

95

95

1500 mg/kg dw

8.0

93

96

94

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

0

2

130

30

10

10

10

0

1

0

1

302

30

10

10

10

3

3

2

8

671

30

10

10

10

4

3

7

14

1500

30

10

10

10

8

8

5

21

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 10.3

30

10

10

10

3

3

3

9

Ref., 18.9

30

10

10

10

8

10

9

27

Ref., 32.0

30

10

10

10

10

9

10

29

The results mean that the 10-d LC50 is 755 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-19

172

755

>1500

302

Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with ant without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
91-98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 130, 302, 671, and 1500 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.26
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
172 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
755 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 500 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
302 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 3.3 (130 mg/kg dw) to 70 % (1500 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 755 mg/kg sediment dw
10-d NOEC = 302 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 130 to 1500 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 130, 302, 671, and 1500 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 % during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

130 mg/kg dw

8.0

93

94

95

302 mg/kg dw

8.1

95

94

95

671 mg/kg dw

8.0

95

95

95

1500 mg/kg dw

8.0

93

96

94

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

0

2

130

30

10

10

10

0

1

0

1

302

30

10

10

10

3

3

2

8

671

30

10

10

10

4

3

7

14

1500

30

10

10

10

8

8

5

21

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 10.3

30

10

10

10

3

3

3

9

Ref., 18.9

30

10

10

10

8

10

9

27

Ref., 32.0

30

10

10

10

10

9

10

29

The results mean that the 10-d LC50 is 755 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-19

172

755

>1500

302

Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
90 - 98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 130, 302, 668, and 1502 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.26
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
362 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
883 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 500 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
302 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 0 (130 mg/kg dw) to 73.3 % (1502 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 883 mg/kg sediment dw
10-d NOEC = 302 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 130 to 1502 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 130, 302, 668, and 1502 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 % during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

130 mg/kg dw

8.1

90

96

97

302 mg/kg dw

8.0

95

95

97

668 mg/kg dw

8.0

96

95

97

1502 mg/kg dw

8.0

93

95

96

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

1

2

130

30

10

10

10

0

0

0

0

302

30

10

10

10

1

0

0

1

668

30

10

10

10

4

4

5

13

1502

30

10

10

10

7

7

8

22

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 9.6

30

10

10

10

3

3

3

9

Ref., 18.6

30

10

10

10

8

10

9

27

Ref., 32.5

30

10

10

10

10

9

10

29

The results mean that the 10-d LC50 is 755 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-20

362

883

>1500

302

Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
90 - 98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 130, 302, 668, and 1502 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.26
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
362 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
883 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 500 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
302 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 0 (130 mg/kg dw) to 73.3 % (1502 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 883 mg/kg sediment dw
10-d NOEC = 302 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 130 to 1502 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 130, 302, 668, and 1502 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 % during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

130 mg/kg dw

8.1

90

96

97

302 mg/kg dw

8.0

95

95

97

668 mg/kg dw

8.0

96

95

97

1502 mg/kg dw

8.0

93

95

96

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

1

2

130

30

10

10

10

0

0

0

0

302

30

10

10

10

1

0

0

1

668

30

10

10

10

4

4

5

13

1502

30

10

10

10

7

7

8

22

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 9.6

30

10

10

10

3

3

3

9

Ref., 18.6

30

10

10

10

8

10

9

27

Ref., 32.5

30

10

10

10

10

9

10

29

The results mean that the 10-d LC50 is 755 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-20

362

883

>1500

302

Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
90 - 98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 129, 300, 671, and 1501 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.27
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
44 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
287 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 501 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
> 129 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 23 (129 mg/kg dw) to 86.7 % (1501 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen content was greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 503 mg/kg sediment dw
10-d NOEC = 126 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 129 to 1501 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 129, 300, 671, and 1501 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was >40% during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

129 mg/kg dw

8.0

97

95

96

300 mg/kg dw

8.0

93

95

96

671 mg/kg dw

8.1

93

94

96

1501 mg/kg dw

8.0

90

93

96

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

1

3

129

30

10

10

10

3

2

2

7

300

30

10

10

10

6

6

7

19

671

30

10

10

10

6

7

7

20

1501

30

10

10

10

9

9

8

26

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 9.6

30

10

10

10

3

3

3

9

Ref., 18.6

30

10

10

10

8

10

9

21

Ref., 32.5

30

10

10

10

10

9

10

29

The results mean that the 10-d LC50 is 287 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-17

44.0

287

>1501

<129

Endpoint:
sediment toxicity: short-term
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
90 - 98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 129, 300, 671, and 1501 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.27
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
44 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
287 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 501 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
> 129 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 23 (129 mg/kg dw) to 86.7 % (1501 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen content was greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 503 mg/kg sediment dw
10-d NOEC = 126 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 129 to 1501 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 129, 300, 671, and 1501 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was >40% during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

129 mg/kg dw

8.0

97

95

96

300 mg/kg dw

8.0

93

95

96

671 mg/kg dw

8.1

93

94

96

1501 mg/kg dw

8.0

90

93

96

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

1

3

129

30

10

10

10

3

2

2

7

300

30

10

10

10

6

6

7

19

671

30

10

10

10

6

7

7

20

1501

30

10

10

10

9

9

8

26

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 9.6

30

10

10

10

3

3

3

9

Ref., 18.6

30

10

10

10

8

10

9

21

Ref., 32.5

30

10

10

10

10

9

10

29

The results mean that the 10-d LC50 is 287 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-17

44.0

287

>1501

<129

Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
90 - 98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 129, 301, 670, and 1501 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.27
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
34 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
332 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 501 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
> 129 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 3 (129 mg/kg dw) to 76.7 % (1501 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 332 mg/kg sediment dw
10-d NOEC < 129 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 129 to 1501 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 129, 301, 670, and 1501 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 %l during the study. The pH was 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

129 mg/kg dw

8.0

90

94

96

301 mg/kg dw

8.1

97

95

97

670 mg/kg dw

8.1

98

94

96

1501 mg/kg dw

8.1

98

93

97

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

1

3

129

30

10

10

10

3

4

2

9

300

30

10

10

10

4

4

5

13

671

30

10

10

10

7

8

7

22

1501

30

10

10

10

7

8

8

23

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 9.6

30

10

10

10

3

3

3

9

Ref., 18.6

30

10

10

10

8

10

9

27

Ref., 32.5

30

10

10

10

10

9

10

29

The results mean that the 10 -d LC50 is 332 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-18

34.0

332

> 1501

< 129

Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 2006-10-18 to 2006-10-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 130 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 110 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 510 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (120 rpm) for 5 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 06.10.2006 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.0 - 8.1
Dissolved oxygen:
90 - 98 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 129, 301, 670, and 1501 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 450 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 9-10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 73%
- Total organic content (%): 3.3


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.27
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
34 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
332 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 501 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
> 129 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 3 (129 mg/kg dw) to 76.7 % (1501 mg/kg dw)
- Mortality in control: 5 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 12.2 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Board (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen contnentwas greater than 40%;

- the 10 -d LC50 for the fluoranthene was 12.2 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 332 mg/kg sediment dw
10-d NOEC < 129 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 129 to 1501 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 129, 301, 670, and 1501 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 %l during the study. The pH was 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.0

96

92

95

129 mg/kg dw

8.0

90

94

96

301 mg/kg dw

8.1

97

95

97

670 mg/kg dw

8.1

98

94

96

1501 mg/kg dw

8.1

98

93

97

Reference, 5.57 mg/kg dw

8.0

98

93

98

Reference, 9.6 mg/kg dw

8.0

97

93

95

Reference, 18.6 mg/kg dw

8.0

91

94

96

Reference, 32.5 mg/kg dw

8.1

96

94

95

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

1

1

1

3

129

30

10

10

10

3

4

2

9

300

30

10

10

10

4

4

5

13

671

30

10

10

10

7

8

7

22

1501

30

10

10

10

7

8

8

23

Ref., 5.57

30

10

10

10

0

0

0

0

Ref., 9.6

30

10

10

10

3

3

3

9

Ref., 18.6

30

10

10

10

8

10

9

27

Ref., 32.5

30

10

10

10

10

9

10

29

The results mean that the 10 -d LC50 is 332 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

7.3

12.2

20.4

5.57

DEV 2006-18

34.0

332

> 1501

< 129

Endpoint:
sediment toxicity: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
from 2008-05-19 to 2008-05-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 73 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 80 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 465 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (110 rpm) for 16 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 17.04.2008 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days (29.09-0.3.10.2006)
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.1
Dissolved oxygen:
93 - 100 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 132, 307, 679, and 1510 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 400 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 70%
- Total organic content (%): 2.5


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.25
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
55 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
1 220 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 500 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
> 132 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 20 (132 mg/kg dw) to 57.7 % (1510 mg/kg dw)
- Mortality in control: 0 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 15.3 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Baord (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen content was greater than 40%;

- the 10 -d LC50 for the fluoranthene was 15.3 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 1220 mg/kg sediment dw
10-d NOEC = <132 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 130 to 1500 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 130, 302, 671, and 1500 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 % during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.1

96

96

98

132 mg/kg dw

8.1

93

96

93

307 mg/kg dw

8.1

96

96

95

679 mg/kg dw

8.1

96

93

96

1510 mg/kg dw

8.1

96

94

97

Reference, 5.4 mg/kg dw

8.1

93

98

98

Reference, 10.5 mg/kg dw

8.1

96

96

100

Reference, 19.1 mg/kg dw

8.1

94

96

93

Reference, 33.8 mg/kg dw

8.1

94

98

97

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

0

0

0

0

130

30

10

10

10

1

4

1

6

307

30

10

10

10

3

3

2

8

679

30

10

10

10

4

3

4

11

1510

30

10

10

10

5

6

6

17

Ref., 5.4

30

10

10

10

0

0

1

1

Ref., 10.5

30

10

10

10

2

2

1

5

Ref., 19.1

30

10

10

10

7

6

6

19

Ref., 33.8

30

10

10

10

10

10

10

30

The results mean that the 10-d LC50 is 1220 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

8.7

15.3

27.0

5.4

EBF 6000

55

1220

>1500

<132

Endpoint:
sediment toxicity: short-term
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Study period:
from 2008-05-19 to 2008-05-30
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good guideline study. However, there is little information on sediment(collection, characteristics...) and organisms (collection, holding and acclimation).
Justification for type of information:
A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: PARCOM Protocols on Methods for the testing of chemicals used in the offshore industry. Part A: A sediment bioassay using an amphipod Corophium sp. (1995)
Deviations:
no
Principles of method if other than guideline:
Guideline study
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested
Analytical monitoring:
not required
Details on sampling:
Not applicable (no analytical monitoring)
Vehicle:
yes
Details on sediment and application:
PREPARATION OF SPIKED SEDIMENT
- Natural sediment from the same unpolluted site as the amphipods (organic content: 3.3%; silt fraction: 73%), homogenized before preparing the test sediment.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone

- Spiked test sediment: 5 mL acetone containing test substance added to 73 g pre-dried sediment. After the acetone had dried off in a fume cupboard, the sample was added to 460 g of damp sediment.

- Reference substance: fluoranthene dissolved in 5 mL of acetone added to app. 80 g pre-dried sediment.After the acetone had dried off in a fume cupboard, the sample was added to app. 465 g of damp sediment in a plastic bucket.

- The sediment in the blanks (with and without acetone) was treated like the test sediment but without adding test substance.

All sediments (test concentrations, reference substance and blanks) were prepared in plastic buckets. 130 mL of seawater was added to each bucket and the bucket were shaken (110 rpm) for 16 hrs.


Test organisms (species):
Corophium volutator
Details on test organisms:
TEST ORGANISM
- Details on collection: collected 17.04.2008 by Biotrix, Norway and transported by car and airplane to the laboratory.
- Size of animals at beginning of exposure: 5-10 mm long
- Feeding during test: no feeding

ACCLIMATION
- Acclimation period: 4 days (29.09-0.3.10.2006)
- Acclimation conditions (same as test or not): temperature 15+/-2 °C
Study type:
laboratory study
Test type:
static
Water media type:
saltwater
Type of sediment:
natural sediment
Limit test:
no
Duration:
10 d
Exposure phase:
total exposure duration
Post exposure observation period:
No post exposure observation
Hardness:
Not relevant (seawater study)
Test temperature:
15 +/- 2°C
pH:
8.1
Dissolved oxygen:
93 - 100 % saturation
Salinity:
34 per thousand
Ammonia:
No data provided
Nominal and measured concentrations:
Test substance nominal concentrations: 132, 307, 679, and 1510 mg/kg dw

Details on test conditions:
TEST SYSTEM
- Test container (material, size): 600 mL glass containers
- Weight of wet sediment: 200 g
- Overlying water volume: 400 mL

EXPOSURE REGIME
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 3
- No. of replicates per control: 3
- No. of replicates per vehicle control: 3


OVERLYING WATER CHARACTERISTCS
- Natural sea water from an unpolluted site at about 80 m depth in Byfjord, filtrated through a sand filter
- Salinity: 34 per thousand


SOURCE OF NATURAL SEDIMENT
- same unpolluted site as the amphipods


CHARACTERIZATION OF SEDIMENT
- Particle size distribution
- % silt: 70%
- Total organic content (%): 2.5


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.25
Reference substance (positive control):
yes
Remarks:
Fluoranthene
Duration:
10 d
Dose descriptor:
LC10
Effect conc.:
55 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
LC50
Effect conc.:
1 220 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
other: LC90
Effect conc.:
> 1 500 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
10 d
Dose descriptor:
NOEC
Effect conc.:
> 132 mg/kg sediment dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Mortality of test animals at end of exposure period: from 20 (132 mg/kg dw) to 57.7 % (1510 mg/kg dw)
- Mortality in control: 0 %
Results with reference substance (positive control):
- Results with reference substance valid
- Relevant effect levels: LC50 = 15.3 mg/kg dw
Reported statistics and error estimates:
LC50, LC10 and LC90 values calculated by using the standard procedure PROBIT analysis, National Swedish Environmental Protection Baord (Version 2.3, 1990).
Calculation of NOEC by single factor ANOVA followed by Dunnett's procedure for comparing each concentation mean with the control mean.

Test acceptability:

The test acceptance criteria given were fulfilled :

- the amphipods were not held without sediment for more than 14 days before they were used for testing;

- the mortality in the individual control replicate did not exceed 40%;

- the entire batch was from the same population;

- the percentage mortality in the blank was less than 15 %;

- the oxygen content was greater than 40%;

- the 10 -d LC50 for the fluoranthene was 15.3 mg/kg.

Validity criteria fulfilled:
yes
Remarks:
Amphipods held without sediment for less than14 days; Mortality in the individual blank less than 40%; All amphipods from the same population; Mortality in the blank less than 15%; 10-d LC50 of reference substance (fluoranthen) = 19.9 +/- 10.3 mg/kg sedim
Conclusions:
10-d LC50 = 1220 mg/kg sediment dw
10-d NOEC = <132 mg/kg sediment dw
Executive summary:

An acute toxicity test was conducted on the marine amphipod Corophium volutator. The test substance concentrations ranged from 130 to 1500 mg/kg sediment dw. The test sediment was from the same unpolluted site as the amphipods. The seawater was natural seawater from an unpolluted site at about 80 m depth in Byfjord, filtered through sand filter at the collection. The test substance was dissolved in acetone and added to the sediment as follows: 5 mL acetone containing the test substance was added to 130 g pre-dried sediment; after the acetone had dried off in a fume cupboard, the sample was added to 510 g of damp sediment in a plastic bucket, 130 mL seawater was added to the bucket and the bucket was shaken (120 rpm) for 5 hrs. The spiked sediment was then split into three beakers, 400 mL of seawater was added to each beaker, and they were aerated overnight before the test organisms were introduced. The blanks were prepared the same way (three with and three without acetone). In the test, 4 concentrations were tested in a geometric series: 130, 302, 671, and 1500 mg/kg. Three replicates were used for each concentration and each blank.

Dissolved oxygen was > 40 % during the study. The pH was 8.0 - 8.1. The temperature was 15 +/- 2 °C.

Test substance concentration

Initial values

Oxygen (% saturation)

pH

O2%

5 d

10 d

Blank

8.1

96

96

98

132 mg/kg dw

8.1

93

96

93

307 mg/kg dw

8.1

96

96

95

679 mg/kg dw

8.1

96

93

96

1510 mg/kg dw

8.1

96

94

97

Reference, 5.4 mg/kg dw

8.1

93

98

98

Reference, 10.5 mg/kg dw

8.1

96

96

100

Reference, 19.1 mg/kg dw

8.1

94

96

93

Reference, 33.8 mg/kg dw

8.1

94

98

97

The test organisms (5 -10 mm) were obtained from the same unpolluted site as the sediment.

No deviation from the protocol was noted. No analytical monitoring was performed.

Results:

Concentration (mg/kg dw)

Total number of specimen per concentration

Number of specimen per vessel

Number of dead

(10 days)

A

D

B

E

C

F

A

D

B

E

C

F

Total

Control

60

10

10

10

10

10

10

0

0

0

0

0

0

0

130

30

10

10

10

1

4

1

6

307

30

10

10

10

3

3

2

8

679

30

10

10

10

4

3

4

11

1510

30

10

10

10

5

6

6

17

Ref., 5.4

30

10

10

10

0

0

1

1

Ref., 10.5

30

10

10

10

2

2

1

5

Ref., 19.1

30

10

10

10

7

6

6

19

Ref., 33.8

30

10

10

10

10

10

10

30

The results mean that the 10-d LC50 is 1220 mg/kg sediment dry weight as shown in the following table:

Test substance

(mg/kg sediment dw)

LC10

LC50

LC90

NOEC

Fluoranthene (reference substance)

8.7

15.3

27.0

5.4

EBF 6000

55

1220

>1500

<132

Endpoint:
sediment toxicity: long-term
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:

Description of key information

The 10d-LC50 and 10d-NOEC for the marine sediment reworker Corophium volutator were reported to be 519 and 100 mg/kg sediment (dw) respectively.

However, as long-term data is not available for this end-point, the environmental risk is assessed using the Petrorisk model.

Key value for chemical safety assessment

Additional information

Short-term studies:

Corophium volutator was exposed to sediment spiked with “Hydrocarbons, C14-C18,n-alkanes, isoalkanes, cyclics, <2% aromatics" for 10 days. Fourteen studies were available and give LC50 values. The geometric mean of LC50 is 519 mg/kg sediment (dw). In most of the studies, LC10 were calculated and the geometric mean of the 12 available values is 112.6 mg/kg sediment (dw). At last, the 7 more recent studies reported NOEC values and the lowest one was 100 mg/kg (dw).

Long-term studies:

No data is available for long-term sediment toxicity.

Substance is a hydrocarbon UVCB. Standard tests for this endpoint are intended for single substances and are not appropriate for this complex substance. However, this endpoint is characterized for representative hydrocarbon structures that comprise the hydrocarbon blocks used to assess the environmental risk of this substance with the PETRORISK model (see library tab in PETRORISK spreadsheet attached in section 13).