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Ecotoxicological information

Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 16 Dec. 2009 to 09 Jan. 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
This study was performed according to OECD Guideline 203 with GLP statement. All validity criteria were fulfilled and no deviations were observed. Therefore, this study is considered reliable without restriction.
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
17th July 1992
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
Samples were taken for analysis from the freshly prepared solutions on the first and third day and also from these solutions after their replacement with freshly prepared solutions ("expired" solutions; after exposure approximately 24h).
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 0.300, 0.405, 0.548, 0.740 and 1.000 g of the test substance were added into the test water directly and then the respective bottles sealed. The test substance was dispensed in 10 L of water with the aid of propeller stirring at approximately 400 rpm for a period of 24h at 23 +/- 2°C and appeared clear and colorless after mixing. After 24h the stirring was stopped to give the test solution.
- Eluate: test water
- Control: yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: zebra fish (Brachydanio rerio)
- Strain: no data
- Source: Qianmin Fishery of Yuhong District in Shenyang
- Age at study initiation (mean and range, SD): juvenile
- Length at study initiation (length definition, mean, range and SD): 2-3 cm
- Weight at study initiation (mean and range, SD): no data
- Method of breeding: no data
- Feeding during test: no

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: no data
- Feeding frequency: daily until 24h before the test was started.
- Health during acclimation (any mortality observed): there were not any signs of disease and mortalities were less than 5% during this acclimatization period.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Remarks on exposure duration:
none
Post exposure observation period:
None
Hardness:
126 mg/L as CaCO3
Test temperature:
21.2-21.6°C
pH:
7.46-7.68
Dissolved oxygen:
7.35-8.48 mg/L
>60% of the air saturation value throughout the test
Salinity:
Not applicable
Conductivity:
No data
Nominal and measured concentrations:
- Nominal concentrations: 30.0, 40.5, 54.8, 74.0 and 100.0 mg/L
- Measured concentrations: 32.0, 41.4, 57.2, 77.8 and 111.7 mg/L in the freshly prepared solutions and 32.1, 36.9, 53.9, 74.2 and 101.2 mg/L in expired solutions. See table 6.1.1/1 in "Any other information on results incl. tables".
Details on test conditions:
TEST SYSTEM
- Test vessel: glass jars
- Type (delete if not applicable): no data
- Material, size, headspace, fill volume: diameter = 20 cm; high = 40 cm; 12.3-12.5 L full of water
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): renewal of the test solutions every 24h.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): not applicable
- Biomass loading rate: < 1.0 g fish/L water

TEST MEDIUM / WATER PARAMETERS
The water used for the aquatic was natural water treated with an activated carbon water filter then aerated for 24h. The physicochemical measurements of the water were: pH was in the range of 6.0 to 8.5, the water hardness was 126 mg/L as CaCO3.

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: 16h light photoperiod daily
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The fish were observed daily. The toxic signs and deaths were recorded at 3, 6, 24, 48, 72 and 96h after exposure. The criterion for death was no opercula movement during a 15 second period of observation.

TEST CONCENTRATIONS
- Range finding study: yes
- Test concentrations: no data
- Results used to determine the conditions for the definitive study: no data
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
53.7 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% CL: 43.7-65.9 mg/L
Duration:
96 h
Dose descriptor:
other: The highest concentration witout observed effects
Effect conc.:
32 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
See table 6.1.1/2 in "Any other information on results incl. tables".
During the 96h exposure period, the fish treater with the test concentration at 40.5 mg/L (geometric mean measured concentration is 39.1 mg/L), and higher concentrations showed visible toxic signs, e.g., lethargy, slowly swimming, and loss of equilibrium.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
The LC50 and 95% confidence limit values were calculated by the Probit method using the geometric mean measured concentration values.

Table 6.1.1/2: Measured concentrations

Nominal concentrations (mg/L)

Measured concentrations (mg/L)

Mean fresh

Mean expired

Overall mean

0-24h

72-96h

Fresh

Expired

Fresh

Expired

Control

30.0

40.5

54.8

74.0

100.0

ND

30.4

41.9

57.2

81.6

111.7

ND

31.1

35.8

55.7

80.9

101.2

ND

33.6

40.9

57.2

74.2

-

ND

33.2

38.0

52.1

68.1

-

ND

32.0

41.4

57.2

77.8

111.7

ND

32.1

36.9

53.9

74.2

101.2

ND

32.0

39.1

55.5

76.0

106.3

Table 6.1.1/1: Cumulative mortality

Nominal concentrations (mg/L)

Geometric mean measured concentrations (mg/L)

Initial population

Time (h)

Cumulative mortality

Abnormalities

Control

-

7

3

6

24

48

72

96

0

0

0

0

0

0

No observed toxic signs

30.0

32.0

7

3

6

24

48

72

96

0

0

0

0

0

0

No observed toxic signs

40.5

39.1

7

3

6

24

48

72

96

0

0

0

1

1

2

Lethargy; slowly swimming; irreversible effect; visible damage

54.8

55.5

7

3

6

24

48

72

96

0

0

0

2

4

4

Lethargy; slowly swimming; irreversible effect; visible damage

74.0

76.0

7

3

6

24

48

72

96

0

0

0

4

6

6

Lethargy; slowly swimming; irreversible effect; visible damage

100.0

106.3

7

3

6

24

48

72

96

0

0

1

7

7

7

Lethargy; slowly swimming; irreversible effect; visible damage

Validity criteria fulfilled:
yes
Conclusions:
96h-LC50 = 53.7 mg/L (95% CL = 43.7 - 65.9 mg/L), based on geometric mean measured concentrations.
Executive summary:

This study was performed according to OECD Guideline 203 with GLP statement, to assess the 96h acute toxicity of the substance to zebra fish (Brachydanio rerio) under semi-static conditions.

Based on the results of a preliminary test, a definitive test was conducted with nominal concentrations of 30.0, 40.5, 54.8, 74.0 and 100.0 mg/L. A concurrent water control group was performed in parallel. The measured test concentrations were determined by GC system. The mean concentrations, calculated using the geometric means of the fresh and expired solutions were 32.0, 39.1, 55.5, 76.0 and 106.3 mg/L which is 97 -107% of the nominal concentrations, and the concentrations of expired solutions were 89 -101% of initial concentrations.

All validity criteria were fulfilled.

The results showed that the 96h-LC50 of the substance to Brachydanio rerio, based on geometric mean measured concentrations in the fresh and expired solutions, was 53.7 mg/L (Probit method) and the 95% confidence limit was 43.7 - 65.9 mg/L. The highest concentration without observed effect after 96 hours was 32.0 mg/L (geometric mean measured concentration).

Description of key information

OECD Guideline 203, GLP, key study, validity 1:
96h-LC50 = 53.7 mg/L (95% CL: 43.7 - 65.9 mg/L)

Key value for chemical safety assessment

LC50 for freshwater fish:
53.7 mg/L

Additional information

One valid key study is available to assess the 96h acute toxicity of the substance to zebra fish (Brachydanio rerio). This study was performed according to OECD Guideline 203 with GLP statement, under semi-static conditions.

The definitive test was conducted with nominal concentrations of 30.0, 40.5, 54.8, 74.0 and 100.0 mg/L. A concurrent water control group was performed in parallel. The measured test concentrations were determined by GC system. The mean concentrations, calculated using the geometric means of the fresh and expired solutions were 32.0, 39.1, 55.5, 76.0 and 106.3 mg/L which is 97 -107% of the nominal concentrations, and the concentrations of expired solutions were 89 -101% of initial concentrations.

All validity criteria were fulfilled.

The results showed that the 96h-LC50 of the substance to Brachydanio rerio, based on geometric mean measured concentrations in the fresh and expired solutions, was 53.7 mg/L (Probit method) and the 95% confidence limit was 43.7 - 65.9 mg/L. The highest concentration without observed effect after 96 hours was 32.0 mg/L (geometric mean measured concentration).