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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22 March 2017 - 23 May 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
17 July 1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
31 May 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)
Version / remarks:
April 1996
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
TOXI-COOP ZRT., 8230 Balatonfüred, Arácsi út 97., (1045 Budapest, Berlini u. 47-49.), Hungary
Analytical monitoring:
yes
Remarks:
reverse phase HPLC method using UV detection
Details on sampling:
The test items content was determined in ISO Medium at the start and at the end of the test. Five parallel samples were taken from the 100 mg/L concentration level and from the control sample.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
The test solution used in the test was prepared by mechanical dispersion without the aid of any solubilising agents. An amount of 0.5 g test item was dissolved in 5000 mL dilution water (ISO Medium) in order to obtain the concentration of 100 mg/L. The test solution was freshly prepared just before introduction of the test animals (start of the experiment).

- Controls: The dilution water (ISO Medium without addition of the test item) was used as untreated control.
- 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: Zebrafish
- Source: Akvárium Magazin Kft. 1222-Budapest, Dévény u. 36, Hungary
- Age at study initiation: juveniles
- Length at study initiation: The body length of each fish were measured at the end of the test in order to check their compliance with the size range recommended for the species by the test guideline (OECD No. 203). The measured values were within the range of 2.50-2.90 cm.
- Weight at study initiation: At the start of the test (on day 0) 10 fish were introduced into aquariums, respectively. The body weight of fish was measured and recorded: first the weight of a vessel filled with water was determined. Then the fish were selected randomly and placed into this vessel. The weight of the group of fish was registered and the average body weight was calculated for 1 fish. The loading of the test aquariums was also calculated and was 0.64 g/L in the control and 0.48 g/L in the test item treated group.
- Method of breeding: Fish are bred in a well-known fish farm, under disease- and parasite-controlled conditions.
ACCLIMATION
- Acclimation period: at least 12 days before test initiation
- Acclimation conditions: same as test
- Type and amount of food during acclimation: fish were fed with appropriate, commercial diet for fish
- Feeding frequency during acclimation: at least three times per week until one day before the test start
- Health during acclimation: No mortality occurred in seven days before the start of the experiment, therefore the batch was considered to be acceptable for testing.

FEEDING DURING TEST
- No feeding during the exposure period.
Test type:
static
Water media type:
freshwater
Remarks:
ISO medium, prepared according to recommendation of Annex 2 of the OECD 203 guideline
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
No post exposure observations were performed.
Hardness:
249 mg/L (as CaCO3).
Test temperature:
21.7 - 22.4 °C
pH:
7.48 - 8.12
Dissolved oxygen:
62.9 – 94.9 % of the air saturation value
Salinity:
NA
Conductivity:
NA
Nominal and measured concentrations:
Nominal concentration: 100 mg/L (limit concentration)
Measured concentration: 100.8 mg/L at the start and 98.9 mg/L at the end of the test
Details on test conditions:
TEST SYSTEM
- Test vessel: aquariums
- Material, size, headspace, fill volume: glass, with approx. 6 litres capacity
- Aeration: Test solutions were not aerated during the test.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The ISO medium was prepared by adding 25 mL from each of four stock solutions to one litre deionised water proportionally.
Stock solution 1: CaCl2 x 2 H2O 11.76 g/L
Stock solution 2: MgSO4 x 7H2O 4.93 g/L
Stock solution 3: NaHCO3 2.59 g/L
Stock solution 4: KCl 0.23 g/L

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light: 8 h dark

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: Toxic effects were not observed during the preliminary test, therefore, a limit test was carried out with one treatment group at a concentration of 100 mg/L (limit concentration) and a concurrent control.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
Behavioural abnormalities:
- Observations on body length and weight: none
- Other biological observations: none
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: none
Reported statistics and error estimates:
Since a limit test was performed (and mortality has not occurred), no statistical analysis was necessary. The LC0, LC50, NOEC, LOEC and the LC100 values were determined directly from the raw data.

Table1: Results from Preliminary Range-Finding Test

 Nominal concentrations [mg/L]  Untreated control  100
 Number of treated animals  3
 Number of dead animals (at 96h)  0  0

Table 2: Summary of the biological endpoints in the main test

Parameter

Nominal concentration
[mg/L]

96-h LC50

> 100

96-h NOEC

100

96-h LC0

100

96-h LOEC

> 100

96-h LC100

> 100

Validity criteria fulfilled:
yes
Conclusions:
The test item had no toxic effect up to at least the limit concentration of 100 mg/L on Zebrafish. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.
Executive summary:

The purpose of this study was to evaluate the acute toxicity of the test item on Zebrafish (Brachydanio rerio). A limit test was performed in which the test animals were exposed to aqueous test media containing the test item for 96 hours at only one concentration (100 mg/L) plus a control in order to demonstrate that the test item is not toxic to Zebrafish up to at least this test concentration (i.e. limit concentration). Based on the results obtained during analytical method validation the test item was expected to be stable for the duration of 96 hours in ISO Medium, therefore, the experiment was carried out under static conditions.

For this purpose, young fish were exposed in a static test to aqueous test media containing the test item for 96 hours. The test was run according to OECD TG 203, EU method C.1 and EPA OPPTS 950.1075. Mortality and any sub-lethal effects were not observed during the 96-h exposure period neither in the treated nor in the control group. Accordingly, the 96-h NOEC was determined as 100 mg/L and the LOEC and the LC50 values were determined to be higher than 100 mg/L.

Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Please refer to section 13 for "Read-Across justification".
Reason / purpose:
read-across source
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality

Description of key information

The calcium salt of the target substance had no toxic effect up to at least the limit concentration of 100 mg/L on Zebrafish. Accordingly, the 96-h LC50 value was determined to be > 100 mg/L. The 96-h NOEC was determined to be 100 mg/L.

Key value for chemical safety assessment

Additional information

The short-term toxicity to fish was addressed with read-across approach to the calcium salt of the target substance.

The purpose of this study was to evaluate the acute toxicity of the analouge substance on Zebrafish (Brachydanio rerio). A limit test was performed in which the test animals were exposed to aqueous test media containing the test item for 96 hours at only one concentration (100 mg/L) plus a control in order to demonstrate that the test item is not toxic to Zebrafish up to at least this test concentration (i.e. limit concentration). Based on the results obtained during analytical method validation the test item was expected to be stable for the duration of 96 hours in ISO Medium, therefore, the experiment was carried out under static conditions.

For this purpose, young fish were exposed in a static test to aqueous test media containing the test item for 96 hours. The test was run according to OECD TG 203, EU method C.1 and EPA OPPTS 950.1075.Mortality and any sub-lethal effects were not observed during the 96-h exposure period neither in the treated nor in the control group. Accordingly, the 96-h NOEC was determined as 100 mg/L and the LOEC and the LC50 values were determined to be higher than 100 mg/L.