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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:
1998-07-27 to 1998-07-31
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Guideline study. The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Nominal loading rates: 0 (Control) and 100 mg/L

- Sampling method: Duplicate samples of freshly-prepared test media from treated and control media were taken at the start. For determination of the stability of the test substance under the test conditions, samples of the test media were taken in duplicate from the treated medium and the control on Day 2 (after 48 hours) and Day 4 (after 96 hours).

The samples were taken by pipette from the approximate centre of the aquaria without further mixing of the media so that the samples only contained dissolved and homogeneously dispersed test substance.

- Sample storage conditions before analysis: Samples were stored refrigerated prior to analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: The test medium was prepared by weighing 300 mg of the test substance into a beaker. Approximately 150 mL of dilution water was added and the suspension was ultrasonically treated for 5 minutes and then stirred. The contents of the beaker were then poured into the test aquarium. The beaker was rinsed a further three times with 150 mL of water and the water added to the aquarium. The aquarium was then topped up to 3 L with dilution water and then stirred for 5 minutes. The test medium was prepared approximately 2 hours before introducing the fish.

- Controls: Dilution water

- Evidence of undissolved material (e.g. precipitate, surface film, etc): The test substance was not fully soluble.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM

- Common name: Zebrafish

- Source: City Zoo, Darmstadt, Germany.

- Length at study initiation (length definition, mean, range and SD): 3.3 +/-0.2 cm

- Weight at study initiation (mean and range, SD): 0.32 +/-0.03 g

- Feeding during test: none

ACCLIMATION

- Acclimation period: 2 weeks

- Acclimation conditions (same as test or not): yes

- Type and amount of food: Tetra Min, TETRA-Werke, Melle, Germany

- Health during acclimation (any mortality observed): 0.7% in 5 weeks prior to test
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
250 mg/L as CaCO3
Test temperature:
22ºC
pH:
7.7-9.7
Dissolved oxygen:
≥7.8 mg/L
Salinity:
not applicable
Nominal and measured concentrations:
Nominal loading rates: 0(Control) and 100 mg/L.

Measured concentrations in the fresh treated test medium were between 65 and 66% of the nominal 100 mg/L loading rate. Over the course of the test the concentration increased to between 80 and 84% of nominal.

The results have been reported and interpreted with reference to nominal loading rates. It is not possible to distinguish between concentrations of the parent substance and its hydrolysis products.
Details on test conditions:
TEST SYSTEM

- Test vessel: aquaria

- Type (delete if not applicable): open

- Material, size, headspace, fill volume: glass, 5 L containing 3 L of test medium.

- Aeration: yes

- Renewal rate of test solution (frequency/flow rate): daily

- No. of organisms per vessel: 7

- No. of vessels per concentration (replicates): 1

- No. of vessels per control (replicates): 1

- Biomass loading rate: 0.75 g/L

TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: Reconstituted freshwater prepared by adding analytical grade salts to deionised water.

- Alkalinity: 0.8 mmol/L

- Ca/mg ratio: 4:1

- Culture medium different from test medium: no

- Intervals of water quality measurement: daily

OTHER TEST CONDITIONS

- Adjustment of pH: no

- Photoperiod: 16 h light, 8 h dark

- Light intensity: 290-550 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality after 2, 24, 48, 72 and 96 h.

TEST CONCENTRATIONS

- Spacing factor for test loading rates: Not applicable - limit test

- Range finding study: yes but results not reported
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
but exposure was to test substance and its hydrolysis products
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
>= 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
but exposure was to test substance and its hydrolysis products
Basis for effect:
mortality (fish)
Details on results:
- Mortality of control: 0

- Effect concentrations exceeding solubility of substance in test medium: The test substance was not fully soluble at all the tested loading rates.
Reported statistics and error estimates:
There were no effects on mortality or other symptoms of toxicity during the test. Statistical analysis of the results was therefore not required.
Sublethal observations / clinical signs:

Table 1. Results of analysis of test media

 

Nominal loading rate (mg/L)

Age of test medium (h)

Measured test substance concentration (mg/L)

Measured concentration as percentage of nominal

0 (Control)

0

<LOD

-

0 (Control)

48

<LOD

-

0 (Control)

96

<LOD

-

100 mg/L

0

65.5

65

100 mg/L

48

79.5

80

100 mg/L

96

84.4

84

100 mg/L

0

65.8

66

100 mg/L

48

80.3

80

100 mg/L

96

84.0

84

 

 Table 2. Test results

 

Nominal loading rate (mg/L)

Percentage mortality after 96 hours

0 (Control)

0

100 (Control)

0

 

Validity criteria fulfilled:
yes
Conclusions:
A 96 hour LL50 value of >100 mg/L and NOELR of ≥100 mg/L have been determined for the effects of the test substance on mortality of Brachydanio rerio. The results are expressed in terms of nominal loading rates of the substance. The test substance hydrolyses in water and it is therefore likely that the test organisms were exposed to a mixture of parent substance and hydrolysis products. The test media contained dissolved and dispersed material.

Description of key information

A 96 hour LL50 value of >100 mg/L and NOELR of ≥100 mg/L (nominal concentration) (highest concentration tested) have been determined for the effects of the test substance on mortality of Brachydanio rerio.

Key value for chemical safety assessment

Additional information

A 96 hour LL50 value of >100 mg/L and NOELR of ≥100 mg/L have been determined for the effects of the test substance on mortality of Brachydanio rerio (highest concentration tested), in compliance with OECD guideline 203. The results are expressed in terms of nominal loading rates of the substance. The test substance hydrolyses in water and it is therefore likely that the test organisms were predominantly exposed to the hydrolysis products. The test media contained dissolved and dispersed material.