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Long-term toxicity to aquatic invertebrates

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Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08 - 23 Aug 1996
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study period of 15 days does not meet the criteria of today standard methods recommending 21 days study duration. However, according to the Guidance on information requirements and chemical safety assessment, Chapter R7.b (ECHA, 2012) the study can be regarded as valid for a long-term evaluation as corresponding criteria were fulfilled (e.g. 80% survival in the control, surviving females produce three broods in 21 days or less, > 60 brood neonates per female in the control on average). Thus all requested endpoints could be determined (time to first brood, number of offspring produced per female, growth and survival) and NOELR values for survival or reproductive endpoints could be calculated.
Qualifier:
according to guideline
Guideline:
other: OECD guideline 202 "Daphnia sp. Acute Immobilization Test and Reproduction Test", Part 2 (1993)
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Deviations:
yes
Remarks:
Only 15 d test duration; Adopted in 1998
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: One set of final and initial WAF samples was collected on day 14 for chemical analysis. WAF samples (initial and final) of the control, the lowest (24 mg/L), middle (242 mg/L), and highest ( 2570 mg/L) were collected in duplicate.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Generation of WAFs was carried out using a modification of the procedure developed by Anderson et al. (1974). Six individual WAF setups, one for each of the five exposure concentrations and one for control were prepared. An appropriate amount of test material was added to 2000 mL of Mobil Technical Center well water in each corresponding WAF aspirator bottle. Bottles were capped with parafilm and stirred for 20 h at room temperature. After stirring, the phases were allowed to separate for 4 h. 800 mL of the water phase were subsequently drained from the bottom spout.
- Differential loading: yes
- Controls: Mobil Technical Center well water without test material.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Source: The primary culture originated from Aquatic Research Organisms, Hampton, NH. The in-house culture had been maintained in the laboratory since June 19, 1996.
- Age of parental stock: Daphnids used for the test were less than 24 h old.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
15 d
Test temperature:
19.3-20.5 °C
pH:
7.84-8.41
Nominal and measured concentrations:
Nominal: 0, 24, 97, 242, 1018 and 2570 mg/L
Measured: Below the detection limit of 0.03 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 mL polystyrene cups with 40 mL of test solution
- Type (delete if not applicable): covered with a plexiglass sheet
- Renewal rate of test solution (frequency/flow rate): every 48 h
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Mobil Technical Center well water
- Intervals of water quality measurement: at the beginning of the study and after every renewal period

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h dark

EFFECT PARAMETERS MEASURED: immobilisation and reproduction
Reference substance (positive control):
no
Duration:
15 d
Dose descriptor:
EL50
Effect conc.:
> 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
immobilisation
Duration:
15 d
Dose descriptor:
EL50
Effect conc.:
> 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
reproduction
Duration:
15 d
Dose descriptor:
NOELR
Effect conc.:
>= 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
immobilisation
Duration:
15 d
Dose descriptor:
NOELR
Effect conc.:
>= 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
reproduction
Details on results:
- Mortality of parent animals: 0%

No effects were observed in the range of water solubility with an NOELR = 2570 mg/L (15 d) based on reproduction and immobilisation.

Table 1: Final total reproduction counts taken during the static renewal three-brood chronic toxicity study of the WAF of the test substance to Daphnia magna

Replicates

Control

Exposure Concentrations (ppm)

24

97

242

1018

2570

A

89

107

102

125

109

112

B

94

113

111

101

135

127

C

115

123

129

120

88

68

D

108

92

92

125

130

109

E

124

122

127

100

123

118

F

117

120

116

121

125

127

G

115

98

0

124

142

129

H

89

79

102

103

128

129

I

114

128

136

102

79

116

J

128

151

140

56

130

116

Totals

1093

1133

1055

1077

1189

1151

Average

109

113

106

108

119

115

Std. Dev.

14

20

40

21

21

18

 

Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02 - 24 Aug 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
adopted in 2008
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Treatment, control
- Sampling method: Samples were taken from each new treatment WAF and the corresponding old solutions (three individual replicates) to determine the WAF concentration stability. The analysis was conducted at 4 different time points for both the new and old solutions during the duration of the test. Samples were collected in duplicate in 30 mL vials with PTFE lined screw cap lids with no headspace.
- Sample storage conditions before analysis: Samples were all analyzed on the day of collection so refrigeration was not necessary.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A water-accommodated fraction (WAF) was tested. The WAF was prepared by adding the appropriate amount of the test substance to the dilution water in glass aspirator bottles using a plastic syringes. The syringes were weighed with and without the test substance to determine the actualloading rate. The vessels were sealed with foil-covered rubber stoppers. The solutions were mixed with Teflon®-coated stirbars on magnetic stirplates. The vortex was set at <10% (actual vortex = 7%) of the static liquid depth. The solutions were mixed for 24 hours (± 1 hour). The solution appeared clear, with bubbled test substance on the surface of the treatment during mixing and after mixing was stopped. At the end of mixing, the solutions were allowed to settle for 1 hour (± 15 minutes). At the end of the settling period, the WAFs were removed from the mixing vessel through the outlet at the bottom of the vessel and placed into 2 L volumetric flasks. 50 µL of Vita chem fresh formula was added to each flask and the test solutions were mixed for an additional 15 minutes prior to the distribution of the WAFs to the test chambers. New WAF solutions were prepared every 48 hours for the renewals.
- Evidence of undissolved material: No undissolved test substance was observed in any of the test chambers.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: Organisms were cultured in the Environmental Toxicology Laboratory, Annandale, USA. The original daphnid culture was supplied by Aquatic Biosystems Inc., Fort Collins, USA.
- Age of parental stock: < 24 h old, taken from 13-day old parents
- Feeding during test
- Food type: Vita-chem fresh water formula (Boyd Enterprises Inc.) and suspension of Pseudokirchneriella subcapitata (Aquatic Biosystems Inc.)
- Amount: Vita-chem fresh water formula: 25 µL/L; Suspension of Pseudokirchneriella subcapitata: 1.3E+08 cells/mL suspension
- Frequency: Vita-chem fresh water formula: Vita-chem feed was added to the treatment WAF before start of the test; Suspension of Pseudokirchneriella subcapitata: 0.35 mL of the suspension were fed at inition of the test and during renewals. Beginning on Day 7, they were fed an additional 0.20 mL suspension on non-renewal days.

ACCLIMATION
- Acclimation conditions: Eight to ten daphnids were kept in 1 L glass culture beakers with approximately 800 mL of reconstituted water. The culture chamber was maintained at 20 ± 1°C under a 16 hour light 8 hour dark photoperiod (10 - 20 foot/candles, 108 - 215 Lux). Two sets of Day 0 cultures were started at least five days a week. Cultures were transferred every other day, with exceptions on holidays or weekends when staff was not present, the brood stock health was evaluated and any mortality, production of males or ephippia was documented as well as any mitigation procedures.
- Type and amount of food: Pseudokirchneriella subcapitata (approximately 4.5 - 6.0E+5 cells/mL). They were also fed 25 µL/L of Vita chem fresh formula mixed on a magnetic stir plate with the reconstituted water prior to feeding with algae.
- Feeding frequency: The culture was fed every other day or as needed based on observed algal clearing.
- Health during acclimation (any mortality observed): The neonates had <10% daily mortality on the experimental start day.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d
Hardness:
170 - 178 mg/L as CaCO3
Test temperature:
20.4 - 22.4 °C
pH:
7.63 - 8.45
Dissolved oxygen:
6.69 - 8.88 mg/L
Nominal and measured concentrations:
Nominal: 135 mg/L loading rate WAF
Details on test conditions:
TEST SYSTEM
- Test vessel: bottles sealed with PTFE-lined caps
- Type: closed
- Material: glass; Size: 130 mL; Fill volume: 130 mL
- Aeration: dilution water was aerated prior to use
- Renewal rate of test solutio: The treatment and the control were renewed at approx. 48 h intervals using the WAF that were prepared approx. 24 h prior to renewal.
- No. of organisms per vessel: 1
- No. of vessels per concentration: 10
- No. of vessels per control: 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: UV-sterilized deionized well water that is treated and distributed throughout the tesing facility via PVC and stainless steel pipes
- Total organic carbon: < 0.05 ppm
- Alkalinity: 117 mg/L as CaCO3
- Intervals of water quality measurement: Water quality measurements (pH, dissolved oxygen, hardness and temperature) were performed on a sub-sample of new WAF, and on old WAF composited from the replicates in each treatment and control twice weekly during the exposure period.

OTHER TEST CONDITIONS
- Photoperiod: 16 hlight: 8 h dark
- Light intensity: Daylight intensity ranged from approximately 131.2 to 169.8 lux during full daylight.

EFFECT PARAMETERS MEASURED: The daphnids were observed daily for immobilization, reproduction and abnormal behavior/appearance (body length) and the number of young were observed daily, but counted upon renewal.
Reference substance (positive control):
no
Duration:
21 d
Dose descriptor:
NOELR
Effect conc.:
>= 135 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
reproduction
Details on results:
- Mortality of parent animals: No mortality observed.
- No. of offspring produced per female: control: 112; treatment: 107
- Body length and weight of parent animals: There were no effects on growth, based on the average adult length measurement of 4.9 mm for the control and 5.0 mm for the treatment.
- Time to first brood release or time to hatch: control: 8 -10 days; treatment: 9 - 10 days

Reported statistics and error estimates:
The NOELR value for reproduction was determined using the Two-sample T-Test method.

TOC analysis confirmed the low water solubility of the test substance in water. The total carbon content of the treatment is only slightly higher than that of the control. Thus, the results of the TOC analysis can not be used for the determination of an effect concentration. Thus, the effect concentration is related to the nominal concentration.

Table 1: Results of TOC analysis [mg/L]

 

New solutions

 

Old solutions

Test day

control

treatment

Test day

control

treatment

0

0.3005

0.4490

2

0.5506

1.0608

8

0.3127

0.3668

10

0.6724

0.4915

14

0.3976

0.4779

16

0.6451

0.7693

20

0.4448

0.5395

21

0.4143

0.4940

Table 2: Survival and reproduction of daphnids during the test in the control

Test day

Number of live offspring released per replicate exposure vessel

Cumulative daphnids immobilized

Percent survival

1

2

3

4

5

6

7

8

9

10

1

0

0

0

0

0

0

0

0

0

0

0

100

2

0

0

0

0

0

0

0

0

0

0

0

100

3

0

0

0

0

0

0

0

0

0

0

0

100

4

0

0

0

0

0

0

0

0

0

0

0

100

5

0

0

0

0

0

0

0

0

0

0

0

100

6

0

0

0

0

0

0

0

0

0

0

0

100

7

0

0

0

0

0

0

0

0

0

0

0

100

8

9

0

0

0

0

0

0

0

0

0

0

100

9

0

0

0

0

0

0

0

0

0

0

0

100

10

14

12

14

8

10

13

16

13

4

15

0

100

11

0

0

0

0

0

0

0

0

0

0

0

100

12

34

0

0

0

17

0

0

0

0

0

0

100

13

0

NC

NC

NC

0

NC

NC

NC

NC

NC

0

100

14

0

26

34

18

0

32

19

27

5

29

0

100

15

NC

0

0

0

NC

0

0

0

0

0

0

100

16

50

40

47

42

54

39

39

0

7

41

0

100

17

0

0

0

0

0

0

0

NC

NC

0

0

100

18

54

0

0

0

0

0

0

39

12

0

0

100

19

0

NC

NC

NC

NC

0

NC

0

0

NC

0

100

20

1

28

23

23

41

28

31

19

55

28

0

100

21

7

0

0

0

0

0

0

0

0

0

0

100

NC: neonates present but not counted

Table 3: Survival and reproduction of daphnids during the test in the treatment

Test day

Number of live offspring released per replicate exposure vessel

Cumulative daphnids immobilized

Percent survival

1

2

3

4

5

6

7

8

9

10

1

0

0

0

0

0

0

0

0

0

0

0

100

2

0

0

0

0

0

0

0

0

0

0

0

100

3

0

0

0

0

0

0

0

0

0

0

0

100

4

0

0

0

0

0

0

0

0

0

0

0

100

5

0

0

0

0

0

0

0

0

0

0

0

100

6

0

0

0

0

*

0

0

0

0

1

90

7

0

0

0

0”

0

*

0

0

0

0

1

90

8

0

0

0

*

0

*

0

0

0

0

2

80

9

0

 NC

0

*

0

*

NC

0

 NC

0

2

80

10

0

8

13

*

16

*

10

7

15

0

2

80

11

0

0

0

*

0

*

0

0

0

0

2

80

12

17

27

24

*

0

*

21

20

0

0

2

80

13

0

0

NC

*

NC

*

0

0

NC

0

2

80

14

0

0

9

*

30

*

0

0

32

28

2

80

15

NC

NC

0

*

0

*

NC

NC

0

0

2

80

16

46

38

44

*

45

*

38

46

43

0

2

80

17

0

0

0

*

0

*

0

0

0

NC

2

80

18

41

34

2

*

1

*

33

0

0

23

2

80

19

NC

NC

NC

*

NC

*

0

NC

NC

0

2

80

20

2

9

31

*

15

*

0

38

17

9

2

80

21

0

0

0

*

0

*

26

0

0

1

2

80

NC: neonates present but not counted

*: adult immobilized

0°: off-color, 0": off color and small

Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
08 - 23 Aug 1996
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: OECD guideline 202 "Daphnia sp. Acute Immobilization Test and Reproduction Test", Part 2 (1993)
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Deviations:
yes
Remarks:
Only 15 d test duration; Adopted in 1998
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: One set of final and initial WAF samples was collected on day 14 for chemical analysis. WAF samples (initial and final) of the control, the lowest (24 mg/L), middle (242 mg/L), and highest ( 2570 mg/L) were collected in duplicate.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Generation of WAFs was carried out using a modification of the procedure developed by Anderson et al. (1974). Six individual WAF setups, one for each of the five exposure concentrations and one for control were prepared. An appropriate amount of test material was added to 2000 mL of Mobil Technical Center well water in each corresponding WAF aspirator bottle. Bottles were capped with parafilm and stirred for 20 h at room temperature. After stirring, the phases were allowed to separate for 4 h. 800 mL of the water phase were subsequently drained from the bottom spout.
- Differential loading: yes
- Controls: Mobil Technical Center well water without test material.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Source: The primary culture originated from Aquatic Research Organisms, Hampton, NH. The in-house culture had been maintained in the laboratory since June 19, 1996.
- Age of parental stock: Daphnids used for the test were less than 24 h old.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
15 d
Test temperature:
19.3-20.5 °C
pH:
7.84-8.41
Nominal and measured concentrations:
Nominal: 0, 24, 97, 242, 1018 and 2570 mg/L
Measured: Below the detection limit of 0.03 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 50 mL polystyrene cups with 40 mL of test solution
- Type (delete if not applicable): covered with a plexiglass sheet
- Renewal rate of test solution (frequency/flow rate): every 48 h
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Mobil Technical Center well water
- Intervals of water quality measurement: at the beginning of the study and after every renewal period

OTHER TEST CONDITIONS
- Photoperiod: 16 h light and 8 h dark

EFFECT PARAMETERS MEASURED: immobilisation and reproduction
Reference substance (positive control):
no
Duration:
15 d
Dose descriptor:
EL50
Effect conc.:
> 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
immobilisation
Duration:
15 d
Dose descriptor:
EL50
Effect conc.:
> 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
reproduction
Duration:
15 d
Dose descriptor:
NOELR
Effect conc.:
>= 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
immobilisation
Duration:
15 d
Dose descriptor:
NOELR
Effect conc.:
>= 2 570 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Water Accommodated Fraction (WAF)
Basis for effect:
reproduction
Details on results:
- Mortality of parent animals: 0%

No effects were observed in the range of water solubility with an NOELR = 2570 mg/L (15 d) based on reproduction and immobilisation.

Table 1: Final total reproduction counts taken during the static renewal three-brood chronic toxicity study of the WAF of the test substance to Daphnia magna

Replicates

Control

Exposure Concentrations (ppm)

24

97

242

1018

2570

A

89

107

102

125

109

112

B

94

113

111

101

135

127

C

115

123

129

120

88

68

D

108

92

92

125

130

109

E

124

122

127

100

123

118

F

117

120

116

121

125

127

G

115

98

0

124

142

129

H

89

79

102

103

128

129

I

114

128

136

102

79

116

J

128

151

140

56

130

116

Totals

1093

1133

1055

1077

1189

1151

Average

109

113

106

108

119

115

Std. Dev.

14

20

40

21

21

18

 

Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
02 - 24 Aug 2011
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
adopted in 2008
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: Treatment, control
- Sampling method: Samples were taken from each new treatment WAF and the corresponding old solutions (three individual replicates) to determine the WAF concentration stability. The analysis was conducted at 4 different time points for both the new and old solutions during the duration of the test. Samples were collected in duplicate in 30 mL vials with PTFE lined screw cap lids with no headspace.
- Sample storage conditions before analysis: Samples were all analyzed on the day of collection so refrigeration was not necessary.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A water-accommodated fraction (WAF) was tested. The WAF was prepared by adding the appropriate amount of the test substance to the dilution water in glass aspirator bottles using a plastic syringes. The syringes were weighed with and without the test substance to determine the actualloading rate. The vessels were sealed with foil-covered rubber stoppers. The solutions were mixed with Teflon®-coated stirbars on magnetic stirplates. The vortex was set at <10% (actual vortex = 7%) of the static liquid depth. The solutions were mixed for 24 hours (± 1 hour). The solution appeared clear, with bubbled test substance on the surface of the treatment during mixing and after mixing was stopped. At the end of mixing, the solutions were allowed to settle for 1 hour (± 15 minutes). At the end of the settling period, the WAFs were removed from the mixing vessel through the outlet at the bottom of the vessel and placed into 2 L volumetric flasks. 50 µL of Vita chem fresh formula was added to each flask and the test solutions were mixed for an additional 15 minutes prior to the distribution of the WAFs to the test chambers. New WAF solutions were prepared every 48 hours for the renewals.
- Evidence of undissolved material: No undissolved test substance was observed in any of the test chambers.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: water flea
- Source: Organisms were cultured in the Environmental Toxicology Laboratory, Annandale, USA. The original daphnid culture was supplied by Aquatic Biosystems Inc., Fort Collins, USA.
- Age of parental stock: < 24 h old, taken from 13-day old parents
- Feeding during test
- Food type: Vita-chem fresh water formula (Boyd Enterprises Inc.) and suspension of Pseudokirchneriella subcapitata (Aquatic Biosystems Inc.)
- Amount: Vita-chem fresh water formula: 25 µL/L; Suspension of Pseudokirchneriella subcapitata: 1.3E+08 cells/mL suspension
- Frequency: Vita-chem fresh water formula: Vita-chem feed was added to the treatment WAF before start of the test; Suspension of Pseudokirchneriella subcapitata: 0.35 mL of the suspension were fed at inition of the test and during renewals. Beginning on Day 7, they were fed an additional 0.20 mL suspension on non-renewal days.

ACCLIMATION
- Acclimation conditions: Eight to ten daphnids were kept in 1 L glass culture beakers with approximately 800 mL of reconstituted water. The culture chamber was maintained at 20 ± 1°C under a 16 hour light 8 hour dark photoperiod (10 - 20 foot/candles, 108 - 215 Lux). Two sets of Day 0 cultures were started at least five days a week. Cultures were transferred every other day, with exceptions on holidays or weekends when staff was not present, the brood stock health was evaluated and any mortality, production of males or ephippia was documented as well as any mitigation procedures.
- Type and amount of food: Pseudokirchneriella subcapitata (approximately 4.5 - 6.0E+5 cells/mL). They were also fed 25 µL/L of Vita chem fresh formula mixed on a magnetic stir plate with the reconstituted water prior to feeding with algae.
- Feeding frequency: The culture was fed every other day or as needed based on observed algal clearing.
- Health during acclimation (any mortality observed): The neonates had <10% daily mortality on the experimental start day.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d
Hardness:
170 - 178 mg/L as CaCO3
Test temperature:
20.4 - 22.4 °C
pH:
7.63 - 8.45
Dissolved oxygen:
6.69 - 8.88 mg/L
Nominal and measured concentrations:
Nominal: 135 mg/L loading rate WAF
Details on test conditions:
TEST SYSTEM
- Test vessel: bottles sealed with PTFE-lined caps
- Type: closed
- Material: glass; Size: 130 mL; Fill volume: 130 mL
- Aeration: dilution water was aerated prior to use
- Renewal rate of test solutio: The treatment and the control were renewed at approx. 48 h intervals using the WAF that were prepared approx. 24 h prior to renewal.
- No. of organisms per vessel: 1
- No. of vessels per concentration: 10
- No. of vessels per control: 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: UV-sterilized deionized well water that is treated and distributed throughout the tesing facility via PVC and stainless steel pipes
- Total organic carbon: < 0.05 ppm
- Alkalinity: 117 mg/L as CaCO3
- Intervals of water quality measurement: Water quality measurements (pH, dissolved oxygen, hardness and temperature) were performed on a sub-sample of new WAF, and on old WAF composited from the replicates in each treatment and control twice weekly during the exposure period.

OTHER TEST CONDITIONS
- Photoperiod: 16 hlight: 8 h dark
- Light intensity: Daylight intensity ranged from approximately 131.2 to 169.8 lux during full daylight.

EFFECT PARAMETERS MEASURED: The daphnids were observed daily for immobilization, reproduction and abnormal behavior/appearance (body length) and the number of young were observed daily, but counted upon renewal.
Reference substance (positive control):
no
Duration:
21 d
Dose descriptor:
NOELR
Effect conc.:
>= 135 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
(WAF)
Basis for effect:
reproduction
Details on results:
- Mortality of parent animals: No mortality observed.
- No. of offspring produced per female: control: 112; treatment: 107
- Body length and weight of parent animals: There were no effects on growth, based on the average adult length measurement of 4.9 mm for the control and 5.0 mm for the treatment.
- Time to first brood release or time to hatch: control: 8 -10 days; treatment: 9 - 10 days

Reported statistics and error estimates:
The NOELR value for reproduction was determined using the Two-sample T-Test method.

TOC analysis confirmed the low water solubility of the test substance in water. The total carbon content of the treatment is only slightly higher than that of the control. Thus, the results of the TOC analysis can not be used for the determination of an effect concentration. Thus, the effect concentration is related to the nominal concentration.

Table 1: Results of TOC analysis [mg/L]

 

New solutions

 

Old solutions

Test day

control

treatment

Test day

control

treatment

0

0.3005

0.4490

2

0.5506

1.0608

8

0.3127

0.3668

10

0.6724

0.4915

14

0.3976

0.4779

16

0.6451

0.7693

20

0.4448

0.5395

21

0.4143

0.4940

Table 2: Survival and reproduction of daphnids during the test in the control

Test day

Number of live offspring released per replicate exposure vessel

Cumulative daphnids immobilized

Percent survival

1

2

3

4

5

6

7

8

9

10

1

0

0

0

0

0

0

0

0

0

0

0

100

2

0

0

0

0

0

0

0

0

0

0

0

100

3

0

0

0

0

0

0

0

0

0

0

0

100

4

0

0

0

0

0

0

0

0

0

0

0

100

5

0

0

0

0

0

0

0

0

0

0

0

100

6

0

0

0

0

0

0

0

0

0

0

0

100

7

0

0

0

0

0

0

0

0

0

0

0

100

8

9

0

0

0

0

0

0

0

0

0

0

100

9

0

0

0

0

0

0

0

0

0

0

0

100

10

14

12

14

8

10

13

16

13

4

15

0

100

11

0

0

0

0

0

0

0

0

0

0

0

100

12

34

0

0

0

17

0

0

0

0

0

0

100

13

0

NC

NC

NC

0

NC

NC

NC

NC

NC

0

100

14

0

26

34

18

0

32

19

27

5

29

0

100

15

NC

0

0

0

NC

0

0

0

0

0

0

100

16

50

40

47

42

54

39

39

0

7

41

0

100

17

0

0

0

0

0

0

0

NC

NC

0

0

100

18

54

0

0

0

0

0

0

39

12

0

0

100

19

0

NC

NC

NC

NC

0

NC

0

0

NC

0

100

20

1

28

23

23

41

28

31

19

55

28

0

100

21

7

0

0

0

0

0

0

0

0

0

0

100

NC: neonates present but not counted

Table 3: Survival and reproduction of daphnids during the test in the treatment

Test day

Number of live offspring released per replicate exposure vessel

Cumulative daphnids immobilized

Percent survival

1

2

3

4

5

6

7

8

9

10

1

0

0

0

0

0

0

0

0

0

0

0

100

2

0

0

0

0

0

0

0

0

0

0

0

100

3

0

0

0

0

0

0

0

0

0

0

0

100

4

0

0

0

0

0

0

0

0

0

0

0

100

5

0

0

0

0

0

0

0

0

0

0

0

100

6

0

0

0

0

*

0

0

0

0

1

90

7

0

0

0

0”

0

*

0

0

0

0

1

90

8

0

0

0

*

0

*

0

0

0

0

2

80

9

0

 NC

0

*

0

*

NC

0

 NC

0

2

80

10

0

8

13

*

16

*

10

7

15

0

2

80

11

0

0

0

*

0

*

0

0

0

0

2

80

12

17

27

24

*

0

*

21

20

0

0

2

80

13

0

0

NC

*

NC

*

0

0

NC

0

2

80

14

0

0

9

*

30

*

0

0

32

28

2

80

15

NC

NC

0

*

0

*

NC

NC

0

0

2

80

16

46

38

44

*

45

*

38

46

43

0

2

80

17

0

0

0

*

0

*

0

0

0

NC

2

80

18

41

34

2

*

1

*

33

0

0

23

2

80

19

NC

NC

NC

*

NC

*

0

NC

NC

0

2

80

20

2

9

31

*

15

*

0

38

17

9

2

80

21

0

0

0

*

0

*

26

0

0

1

2

80

NC: neonates present but not counted

*: adult immobilized

0°: off-color, 0": off color and small

Description of key information

Key value for chemical safety assessment

Additional information

Since no studies investigating the long-term toxicity of Pentaerythritol, mixed esters with linear and branched fatty acids to aquatic invertebrates are available, in accordance to Regulation (EC) No 1907/2006 Annex XI, 1.5 a read-across to the structurally related source substances Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9) and Decanoic acid, ester with 2-ethyl-2-(hydroxymethyl)-1,3-propanediol octanoate (CAS 11138-60-6) was conducted. The target substance is characterized as a tetraester of pentaerythritol and linear and branched fatty acids. The source substances are structurally very similar to the target substance. Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid is a tetraester of C5/C7/C9/C10 with pentaerythritol. The source substance Fatty acids, C8-10 (even numbered), di-and triesters with propylidynetrimethanol (CAS 11138-60-6) is a triester of trimethylolpropane and C8-C10 fatty acids (even numbered).

This read-across is justified in detail in the overall summary (IUCLID Section 6.1) and within the analogue justification in IUCLID Section 13.

The first study on the long-term toxicity to aquatic invertebrates is available for fatty acids, C8-10 (even numbered), di-and triesters with propylidynetrimethanol (CAS 11138-60-6). The study was performed according to OECD 202 "Daphnia sp. Acute Immobilization Test and Reproduction Test", Part 2 (1993) under semi-static conditions (Low, 1996). This test guideline is similar to OECD 211 which was adopted in 1998. The study period of 15 days does not meet the criteria of today standard methods recommending 21 days study duration. However, according to the Guidance on information requirements and chemical safety assessment, Chapter R7.b (ECHA, 2012) the study can be regarded as valid for a long-term evaluation as corresponding criteria were fulfilled (e.g. 80% survival in the control, surviving females produce three broods in 21 days or less, > 60 brood neonates per female in the control on average). In addition, all requested endpoints could be determined (time to first brood, number of offspring produced per female, growth and survival) and NOELR values for survival or reproductive endpoints could be calculated. In this study Daphnia magna were exposed to nominal loading rates of 0, 24, 97, 242, 1018 and 2570 mg/L (prepared as water accommodated fraction, WAF). No effects were observed up to the highest test concentration resulting in a NOELR of2570 mg/L based on reproduction and immobilization.

The study with the source substance decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid (CAS 71010-76-9) investigated the long-term toxicity aquatic invertebrates (Letinski & Bragin, 2011). The GLP study was conducted under semi-static conditions according to OECD 211 using Daphnia magna as test organism. A loading rate of 135 mg/L (nominal) was applied in the limit test. The WAF solution was prepared by adding the appropriate amount of test substance with subsequent stirring and sampling of the aqueous portions (WAFs) through the outlet at the bottom of the vessels. No significant effect on reproduction was observed in the treatment throughout the test period of 21 d. Hence, the NOELR (21 d) is determined to be135 mg/L on the basis of the nominal loading rate.

Based on the available results from structurally related source substances (in accordance to Regulation (EC) No 1907/2006 Annex XI, 1.5) which are characterized by a similar ecotoxicological profile and comparable structure, it can be concluded that Pentaerythritol, mixed esters with linear and branched fatty acids will not exhibit long-term effects to aquatic invertebrates up to the limit of water solubility.