Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
December 5, 2013 - March 27, 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

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
Test material form:
solid
Specific details on test material used for the study:
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test medium (reconstituted water and test item) was prepared freshly. Therefore, the calibrated flask with test medium was treated in an ultrasonic device for 1 hour. Subsequently, the preparation was stirred with a magnetic stirrer for further 23 hours. Afterwards, the formulation was passed through a membrane filter with a pore size of 0.2 µm. The filtrate was used for the study.

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
yes
Details on test solutions:
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test medium (reconstituted water and test item) was prepared freshly. Therefore, the calibrated flask with test medium was treated in an ultrasonic device for 1 hour. Subsequently, the preparation was stirred with a magnetic stirrer for further 23 hours. Afterwards, the formulation was passed through a membrane filter with a pore size of 0.2 µm. The filtrate was used for the study.

VEHICLE:
For the reconstituted water the following chemicals (analytical grade) were dissolved in fully demineralized water:

Macro nutrients mg/L
CaCl2 x 2H2O 294.00
MgSO4 x 7H2O 123.25
NaHCO3 64.75
KCl 5.75

Before the start of the experimental part the reconstituted water for fish was prepared. After preparation the water was aerated for 24 hours before using it.
The proportion of Ca to Mg ions was 4:1 and that of Na to K ions 10:1. Hardness: about 250 mg/L CaCO3.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish
- Strain: Dania rerio
- Source: Breeder is Institute of Toxicology, Merck KGaA
- Length at study initiation: 2.0 + 1.0 cm
- Weight at study initiation: Body weight per group: 5.7 g (control group) and 5.8 g (test item group)
- Feeding during test: commercial fish diet during acclimatization

ACCLIMATION
- Acclimation period: The fish were acclimatized to the conditions of the laboratory for more than 14 days and were held in reconstituted water for at least 12 days before testing.
- Acclimation conditions (same as test or not): same
- Health during acclimation (any mortality observed): no

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 250 mg/L CaCO3
Test temperature:
19.8 -20.6°C
pH:
7.26 - 7.73
Dissolved oxygen:
72.8 - 93.1%
Salinity:
no data
Nominal and measured concentrations:
Nominal Concentration: 100 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 10L Glass Aquariums
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: Full glas folled with 10L test medium
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): static
- 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: reconstituted watr according OECD 203
- Total organic carbon: no data
- Particulate matter: no data
- Metals: no data
- Pesticides:no data
- Chlorine: no data
- Alkalinity: no data
- 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: 12hrs light / 12hrs dark
- Light intensity: no data

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality, clinical symptoms

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

Results and discussion

Effect concentrations
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: > 0.00022 mg/L
Details on results:
The 96 hour LC50 for the test material to Zebrafish could not be quantified due to the absence of toxicity at the highest test concentration achievable in water. An aqueous filtrate of 0.1 g/L nominally revealed no aquatic toxicity in the test system.


The 96h LC50 exceeded the water solubility of 0.00022 mg/L (nominal >100 mg/L) and, thus, could not be determined in this study.


- Behavioural abnormalities: no
- Observations on body length and weight: no
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- 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:
Under the conditions of the present study, an aqueous solution of 100 mg/L of the test item revealed no aquatic toxicity in the test system.The 96h EC50was >0.00022 mg/L (nominal >100 mg/L) and, thus, could not be determined in this study.
Executive summary:

The test was performed according to OECD 203. Under the conditions of the present study, an aqueous solution of 100 mg/L of the test item revealed no aquatic toxicity in the test system.The 96h EC50was >0.00022 mg/L (nominal >100 mg/L) and, thus, could not be determined in this study.