Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 11, 2009 - June 19, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2009
Report Date:
2009

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Version / remarks:
Council Regulation (EC) No. 440/2008 laying down test methods pursuant to Regulation (EC) No. 1907/2006 of the European Parliament and the council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Sampling and analysis

Analytical monitoring:
no
Remarks:
The test item concentration was not quantified at the start and the end of this study. Because of the low calculated water solubility (0.0002214 µg/L), the compound cannot be detected with standard analytical methods.

Test solutions

Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Before exposure of the fish, the test medium (reconstituted water and test material) was freshly prepared. The calibrated flask with test material and vehicle (reconstituted water) was treated in an ultrasonic device for 1 hour. Subsequently, the preparation was aerated and stirred with a magnetic stirrer for further 23 hours. After 24 hours the formulation was given through a nutsch filter (pore size 10 - 16 µm). The filtrate was used for the study.
- Controls: reconstituted water
- Chemical name of vehicle: OECD 203 Medium

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish (Danio rerio)
- Strain: West-Aquarium
- Source: Institute of Toxicology, Merck KGaA
- Age at study initiation (mean and range, SD):
- Length at study initiation: 2.0 ± 1.0 cm
- Weight at study initiation: Body weight per group: 7.1 g (control group) and 7.3 g (test material group)


ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food during acclimation:
- Feeding frequency during acclimation: During the acclimatization, until one day before the experimental part, the fish were fed with flakes (Tetra Min® diet; Tetra GmbH, Melle). Furthermore, the fish were fed with daphnia, or artemia, or frozen mosquitolarva daily.
- Health during acclimation (any mortality observed): no mortality observed


FEEDING DURING TEST
The zebrafish were not fed during the study.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
no

Test conditions

Hardness:
about 267 mg/L CaCO3
Test temperature:
23 to 24°C
pH:
7.07 - 7.95
Dissolved oxygen:
61.3 - 92.3%
Salinity:
no data
Conductivity:
no data
Nominal and measured concentrations:
Nominal Concentration: 100 mg/L
Details on test conditions:

TEST SYSTEM
- Test vessel: 10L glass aquarium
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass, 10L, no headspace, about 10L
- Aeration: none
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water (OECD 203 Medium)
- 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: 12 hours light - 12 hours dark regime
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality and Clinical Symptoms (daily)

TEST CONCENTRATIONS
- Range finding study:
- Test concentrations: 0 mg/L and 100 mg/L (nominal)
- Results used to determine the conditions for the definitive study: no mortality observed
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: > 0.0002214 µg/L
Key result
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: > 0.0002214 µg/L
Key result
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: > 0.0002214 µg/L
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
Remarks on result:
other: > 0.0002214 µg/L
Details on results:
A saturated aqueous solution of Art. 281322 of nominal 100 mg/L revealed no aquatic toxicity in the test system. The 96 h LC50 value to zebrafish exceeded the maximal solubility of 0.0002214 µg/L of the test material in reconstituted water and, thus, could not be determined in this test.


The 96h EC50 exceeded the water solubility of 0.0002214 µg/L (nominal >100 mg/L) and, thus, could not be determined in this study.


- Behavioural abnormalities: no
- 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: no
- Effect concentrations exceeding solubility of substance in test medium: yes

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
An aqueous solution of the test item with a nominal concentration of 100 mg/L revealed no aquatic toxicity in the test system. The 96 h LC50 value for zebrafish exceeded the maximal solubility of the test material in reconstituted water and, thus, could not be determined in this test.
Executive summary:

This study was performed according to OECD 203 under GLP conditions. An aqueous solution of the test item with a nominal concentration of 100 mg/L revealed no aquatic toxicity in the test system. The 96 h LC50value for zebrafish exceeded the maximal solubility of the test material in reconstituted water and, thus, could not be determined in this test.