Registration Dossier

Diss Factsheets

Ecotoxicological information

Long-term toxicity to fish

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
fish early-life stage toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 January - 10 September 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study was conducted in accordance with International guidelines and in accordance with GLP. All guideline validity criteria were met.
Qualifier:
according to guideline
Guideline:
OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
RADIOLABELLING INFORMATION (if applicable)
- Radiochemical purity: N/A
- Specific activity: N/A
- Locations of the label: N/A
- Expiration date of radiochemical substance: N/A

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in a closed container.
- Stability under test conditions: Asumed stable.
- Solubility and stability of the test substance in the solvent/vehicle: Acetone stock solution prepared at 25 mg/mL
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: N/A

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: N/A
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: N/A
- Final preparation of a solid: N/A

FORM AS APPLIED IN THE TEST (if different from that of starting material): Solubilised in an acetone stock solution which further diluted with acetone and spiked into dilution water. Co-solvent was maintained <0.1 %v/v in the test media.

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable): N/A

OTHER SPECIFICS: No
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 0.65, 1.3 and 2.6 mg/L nominal
- Sampling method: Aliquots of the fresh and expired solutions from Day 7, 13, 19 and 28 were taken for analysis. Samples were diluted 1:1 with acetonitrile. Samples from the 1.3 and 2.6 mg/L groups were further diluted 2.5 and 5 times with 50 % acetonitrile (aq.) and then filtered (0.22 µm) in preparation for LC-MS analysis.
- Sample storage conditions before analysis: Not reported
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A 25 mg/mL acetone stock solution was prepared by dispensing 0.25917 - 0.26192 gtest item and dissolving in 10 mL acetone. Aliquots of the stock were diluted (in acetone) to prepare a further 2 stock solutions with a concentration of 6.25 and 12.5 mg/mL. 0.2 mL of each stock was dispensed into 2 L of test medium to prepare the treatment solutions. The solutions were stirred for 2 h.
- Eluate: Demineralised water which was groundwater filtered with activated charcoal and softened with "special techniques".
- Controls: Acetone solvent control run in parallel with treatment solutions/ test groups.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): 0.01 % v/v
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Strain: Danio rerio
- Source: Not reported

METHOD FOR PREPARATION AND COLLECTION OF FERTILIZED EGGS
- Numbers of parental fish (i.e. of females used to provide required number of eggs): Not reported
- Method of collection of fertilised eggs: Not reported
- Subsequent handling of eggs: 60 fertilised eggs were used for each concentration and divided equally into 2 replicate chambers.

POST-HATCH FEEDING
- Start date: Not reported
- Type/source of feed: Yolk and fish feed powder was fed to newly hatched fry and 48 h old brine shrimp nauplii and fish feed powder for juveniles.
- Amount given: Not reported
- Frequency of feeding: ad libitum
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Hardness:
96 - 104 (units not reported)
Test temperature:
24.2 - 25.6 ºC
pH:
7.87 - 8.16
Dissolved oxygen:
72.8 - 120.4 % ASV
Salinity:
N/A
Conductivity:
Not reported
Nominal and measured concentrations:
Nominal: 0.65, 1.3 and 2.6 mg/L
Geometric mean measured: 0.537, 1.24 and 2.25 mg/L
Details on test conditions:
TEST SYSTEM
- Emybro cups (if used, type/material, size, fill volume): 250 mL and 1 L glass beakers filled to 200 mL
- Test vessel: 250 mL and 1 L glass beakers filled to 200 mL
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 250 mL and 1 L glass beakers filled to 200 mL
- Aeration: Not reported
- Type of flow-through (e.g. peristaltic or proportional diluter): N/A, semi-static test
- Renewal rate of test solution (frequency/flow rate): Test solutions renewed every 24 h
- No. of fertilized eggs/embryos per vessel: 30
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 0
- No. of vessels per vehicle control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Demineralised water which was groundwater filtered with activated charcoal and softened by "special techniques".
- Total organic carbon: Not reproted
- Particulate matter: Not reproted
- Metals: Not reproted
- Pesticides: Not reproted
- Chlorine: Not reproted
- Alkalinity: Not reproted
- Ca/mg ratio: Not reproted
- Salinity: Not reproted
- Culture medium different from test medium: No
- Intervals of water quality measurement: Total hardness and pH were measured in all vessels at the beginning and end of the test. DO and temperature were measured twice weekly with temperature also continuously monitored in one vessel.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 12: 12 light: dark
- Light intensity: Not reported

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

Embryonic development - verified as precisely as possible.
Hatching and survival - observed once daily and numbers recorded (including eggs, embryos, larvae and juvenile fish).
Abnormal appearance - recorded at adequate time intervals.
Length and weight - recorded at the end of the test.

VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
- Test concentrations: No
- Results used to determine the conditions for the definitive study: No

POST-HATCH DETAILS
- Begin of post-hatch period: Day 4 = Day 0 post-hatch i.e. 90 % control embryos (and in all treatment groups) hatched by Day 4
- No. of hatched eggs (alevins)/treatment released to the test chamber: N/A embryo cups not utilised.
- Release of alevins from incubation cups to test chamber on day no.: N/A, embryo cups not utilised.

FERTILIZATION SUCCESS STUDY
- Number of eggs used: 60 eggs used per concentration
- Removal of eggs to check the embryonic development on day no.: Not conducted
Reference substance (positive control):
no
Key result
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
2.25 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
other: mortality and sub-lethal effects
Details on results:
- Mortality/survival at embryo, larval, juvenile, and adult stages: No significant differences were noted between the control and treatment groups.
- Days to hatch or time to release of young: No significant differences were noted between the control and treatment groups.
- Numbers hatched, Numbers of offspring produced, or Number of offspring per live female per day: No significant differences were noted between the control and treatment groups.
- Number of fish in swim-up stage at one or more time periods (e.g., day x1, x2): Not reported
- Observations on body length and weight of young and/or exposed parents at one or more time periods: No significant differences were noted between the control and treatment groups.
- Number of healthy fish at end of test: No significant differences were noted between the control and treatment groups.
- Type of and number with morphological abnormalities: No significant differences were noted between the control and treatment groups.
- Type of and number with behavioural abnormalities:No significant differences were noted between the control and treatment groups.
- Type and number of developmental effects: No significant differences were noted between the control and treatment groups.
- Type and magnitude of hormonal changes: Not determined
- Other biological observations: None
- Effect concentrations exceeding solubility of substance in test medium: No
- Incidents in the course of the test which might have influenced the results: No
Reported statistics and error estimates:
The following parameters were available for statistical analysis; cummulative mortality, healthy number of larvae at the end of the test, time to start of hatching and end of hatching (90 % hatched), numbers of larvae hatching each day, length and weight of survivors at test end.

Analysis of variance procedures were used for calculating the variations. Multiple comparisons between the results at the individual concentrations and those for the controls were made.

Table 1         Mean measured concentrations

 

Nominal concentration

(mg/L)

Geometric mean measured concentration

(mg/L)

Component 1

Component 2

Component 3

Component 4

Test item

% of nominal

Control

nd

nd

nd

nd

nd

nd

Solvent control

nd

nd

nd

nd

nd

nd

0.65

0.0582

0.171

0.202

0.105

0.537

82.5

1.3

0.116

0.402

0.476

0.249

1.24

95.4

2.6

0.226

0.721

0.868

0.430

2.25

86.3

nd: not detected

 

Table 2       Number of larvae hatched each day

 

Nominal concentration

(mg/L)

Mean measured concentrations

(mg/L)

Rep.

Initial egg number

Day 1 pre-hatch

Day 2 pre-hatch

Day 3 pre-hatch

Day 0 post-hatch

Day 1 post-hatch

Control

nd

1

30

0

3

13

28

30

2

30

0

1

13

29

30

Solvent control

nd

1

30

0

1

19

29

30

2

30

0

2

10

29

30

0.65

0.537

1

30

0

1

14

30

30

2

30

0

1

15

30

30

1.3

1.24

1

30

0

1

21

30

30

2

30

0

1

24

28

30

2.6

2.25

1

30

0

2

15

30

30

2

30

0

1

15

30

30

nd: not detected

 

Table 3       Cummulative mortality

 

Test phase

Nominal concentration

(mg/L)

Mortality and symptoms

Control

Solvent control

0.65

1.3

2.6

Measured concentration

(mg/L)

nd

nd

0.537

1.24

2.25

Initial numbers

1

2

1

2

1

2

1

2

1

2

Day 1 pre-hatch

30

5

A

6

A

7

D

5

A

6

A

5

A

6

D

6

D

7

A

9

A

Day 2 pre-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

6

D

6

D

7

A

9

A

Day 3 pre-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

D

6

D

7

A

9

A

Day 0 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 1 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 2 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 3 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 4 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 5 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 6 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 7 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 8 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 9 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 10 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 11 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 12 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 13 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 14 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 15 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 16 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 17 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 18 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 19 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 20 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 21 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 22 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 23 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 24 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 25 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 26 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 27 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 28 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 29 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Day 30 post-hatch

30

5

A

6

A

7

A

5

A

6

A

6

A

7

A

6

A

7

A

9

A

Survived fish

30

25

24

23

25

24

24

23

24

23

21

Mortality (%)

30

16.7

20.0

23.3

16.7

20.0

20.0

23.3

20.0

23.3

30.0

Survival rate (%)

30

83.3

80.0

76.7

83.3

80.0

80.0

76.7

80.0

76.7

70.0

P0.05(2-side)

-

-

0.795

0.500

0.500

0.126

Table 4       Weight of surviving animals at the end of the test

 

Nominal concentration

(mg/L)

Measured concentration

(mg/L)

Replicate

Surviving numbers

Total body weight

(mg)

Mean body weight

(mg)

Mean ± SD

(mg)

P0.05

(2-side)

Control

nd

1

25

58.3

2.33

2.37 ± 0.057

-

2

24

57.9

2.41

Solvent control

nd

1

23

53.4

2.32

2.34 ± 0.021

0.39

2

25

58.8

2.35

0.65

0.536

1

24

57.4

2.39

2.37 ± 0.032

0.96

2

24

56.3

2.35

1.3

1.24

1

23

51.7

2.25

2.23 ± 0.022

0.24

2

24

53.2

2.22

2.6

2.25

1

23

48.8

2.12

2.14 ± 0.025

0.061

2

21

45.3

2.16

 

Validity criteria fulfilled:
yes
Conclusions:
There were no significant differences between the control vessels and the treatment groups. The following parameters were observed; mortality and sub-lethal effects (including: number of larvae hatched; time to hatching; number of healthy larvae; length and weight of larvae). The NOEC was therefore estimated as 2.25 mg/L, the highest concentration tested.
Executive summary:

OECD 210 (2013) - The 28–day chronic toxicity of Tetraesters of pentaerythritol with 2-methylpropanoic acid and 3,5,5-trimethyl-hexanoic acid to early life stage of zebra fish (Danio rerio) was studied under semi-static conditions.  Fertilized eggs/embryos, 30 per replicate vessel, of zebra fish were exposed to control, solvent control (acetone, 0.01 % v/v), and test chemical geometric mean measured concentrations of 0.537, 1.24 and 2.25 mg/L.  The test system was maintained at 24.2 – 25.6 ºC and a pH of 7.87 to 8.16. 

 

There were no treatment related effects observed during the test.

 

The 28-day NOEC value, based on mortality/sublethal effects, was 2.25 mg/L, the highest concentration tested.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for early life toxicity study with fish.

Description of key information

Mortality and sub-lethal effects: NOEC = 2.25 mg/L; OECD 210; Hairong, Y. (2013)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
2.25 mg/L

Additional information

OECD 210 (2013) - The 28–day chronic toxicity of Tetraesters of pentaerythritol with 2-methylpropanoic acid and 3,5,5-trimethyl-hexanoic acid to early life stage of zebra fish (Danio rerio) was studied under semi-static conditions.  Fertilized eggs/embryos, 30 per replicate vessel, of zebra fish were exposed to control, solvent control (acetone, 0.01 % v/v), and test chemical geometric mean measured concentrations of 0.537, 1.24 and 2.25 mg/L.  The test system was maintained at 24.2 – 25.6 ºC and a pH of 7.87 to 8.16. 

 

There were no treatment related effects observed during the test.

 

The 28-day NOEC value, based on mortality/sublethal effects, was 2.25 mg/L, the highest concentration tested.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for early life toxicity study with fish.

This toxicity study is classified as acceptable and satisfies the guideline requirement for early life toxicity study with fish.

Categories Display