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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:
06 August 2019 - 18 September 2019
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:
adopted 18 June 2019
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
May 30, 2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)
Version / remarks:
October 2016
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
Batch No.: I1900071-19158-AKM
Purity: 99.8 % (aldimine group contents via acid titration)
Expiry Date: April 25, 2022
Analytical monitoring:
yes
Details on sampling:
For determination of the test item concentration samples were taken from the test solution (5×1 mL) and control (3×1 mL) at the start and at the end of the test.
Vehicle:
no
Details on test solutions:
The test solution used in the test was prepared by mechanical dispersion without using of any solubilising agent. An amount of 0.5093 g test item was dissolved in 5093 mL dilution water (ISO medium) in order to obtain the concentration of 100 mg/L. After the formulation procedure the test animals were immediately introduced into test solution.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish
- Source: Akvárium magazin Kft. (Pasaréti Gyula) 1222 Budapest, Dévény u. 36.
- Age at study initiation: juveniles
- Length at study initiation: 1.4 -1.90 cm
- Weight at study initiation: 0.10 - 0.17 g

ACCLIMATION
- Acclimation period: 9 days
- Acclimation conditions: same as test conditions
- Type of food during acclimation: 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 observed

FEEDING DURING TEST
The fish were not fed during the test.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
249 mg/L (as CaCO3)
Test temperature:
21.1 - 21.6 °C
pH:
7.55 - 9.61
Dissolved oxygen:
81.2 - 86.0 %
Nominal and measured concentrations:
Nominal concentration: 100 mg/L
Measured concentration: 100 mg/L (0h), 98.6 mg/L (96h)
Details on test conditions:
TEST SYSTEM
- Test vessel: aquarium
- Type: open
- Material, fill volume: glass, 5L
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.20 g/L in the control and 0.34 g/L in the test item treated group

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: ISO medium according to OECD 203

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light : 8 h dark
- Light intensity: 707 lux

EFFECT PARAMETERS MEASURED
As a limit test was performed and no toxicity was expected observation of fish was sufficient at least once daily. Fish were observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality.

TEST CONCENTRATIONS
Range finding study :
- Test concentrations: 100 mg/L
- Results used to determine the conditions for the definitive study: yes, no mortality was oberved.
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
Details on results:
Validity of the Study
- No mortality was observed in the control group during the test.
- The dissolved oxygen concentration in the test solutions did not fall below 60 % of air saturation value during the study.
- Test concentration was analytically determined.
All acceptance criteria were met therefore the study was considered to be valid.
Reported statistics and error estimates:
Statistical analysis was not necessary; endpoints were determined directly from the raw data.
Validity criteria fulfilled:
yes
Conclusions:
In the 96-hour static acute toxicity test on Zebrafish (Danio rerio) the test item had no toxic effect on fish at the limit concentration of 100 mg/L. 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 is to evaluate the acute toxicity of the test item on Zebrafish (Danio rerio; formerly known as 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 the limit test concentration of 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). The test method of application and the test species Zebrafish (Danio rerio) are recommended by the test guidelines. Fish were exposed to aqueous test media containing the test item for 96 hours in a static test. Ten fish were exposed to the test concentration or run as control (no replicates were used). Fish were observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen and temperature were carried out daily. Concentrations were determined from the treatment and control group sampled at the start (0 hour) and at the end of exposure (96 hours) using HPLC-UV method. The mean measured concentration of the test item was 100 mg/L at the start and 98.6 mg/L at the end of the test. The measured test item concentration remained within ± 20 % of the nominal concentration over the test period of 96 hours, therefore the biological results are based on the nominal concentration. All validity criteria were met and therefore the study was considered to be valid. Mortality and any sub-lethal effects were not observed during the 96-h exposure period either in the treated or in the control group. Accordingly, the 96-h NOEC was determined to be 100 mg/L. The LOEC and the LC50 values were determined to be higher than 100 mg/L.

Description of key information

In the 96-hour static acute toxicity test on Zebrafish (Danio rerio) the test item had no toxic effect on fish at the limit concentration of 100 mg/L. 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 purpose of this study is to evaluate the acute toxicity of the test item on Zebrafish (Danio rerio; formerly known as 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 the limit test concentration of 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). The test method of application and the test species Zebrafish (Danio rerio) are recommended by the test guidelines. Fish were exposed to aqueous test media containing the test item for 96 hours in a static test. Ten fish were exposed to the test concentration or run as control (no replicates were used). Fish were observed at approximately 24, 48, 72 and 96 hours after start of the test for signs of intoxication and mortality. Measurements of pH, dissolved oxygen and temperature were carried out daily. Concentrations were determined from the treatment and control group sampled at the start (0 hour) and at the end of exposure (96 hours) using HPLC-UV method. The mean measured concentration of the test item was 100 mg/L at the start and 98.6 mg/L at the end of the test. The measured test item concentration remained within ± 20 % of the nominal concentration over the test period of 96 hours, therefore the biological results are based on the nominal concentration. All validity criteria were met and therefore the study was considered to be valid. Mortality and any sub-lethal effects were not observed during the 96-h exposure period either in the treated or in the control group. Accordingly, the 96-h NOEC was determined to be 100 mg/L. The LOEC and the LC50 values were determined to be higher than 100 mg/L.