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Ecotoxicological information

Long-term toxicity to aquatic invertebrates

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Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
adopted in 2008
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid
EC Number:
275-118-0
EC Name:
Decanoic acid, mixed esters with heptanoic acid, octanoic acid, pentaerythritol and valeric acid
Cas Number:
71010-76-9
IUPAC Name:
71010-76-9

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Concentrations: Treatment, control
- Sampling method: Samples were taken from each fresh treatment WAF and the corresponding old solutions (three individual replicates) to determine the Water Accomodated Fraction (WAF) concentration stability. The analysis was conducted at 4 different time points for both the fresh 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.

Test solutions

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. Fresh 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

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: The neonates had <10% daily immobilisation on the experimental start day.

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d

Test conditions

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 fresh 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 light: 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

Results and discussion

Effect concentrations
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:
- Immobilisation of parent animals: No immobilisation 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.

Any other information on results incl. tables

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]

 

Fresh 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

Applicant's summary and conclusion