Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 2, 2013 - December 31, 2013
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
No European GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
1992
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
4-(4-butylphenyl)-4'-ethyl-3-fluoro-1,1'-biphenyl
EC Number:
617-357-9
Cas Number:
825633-75-8
Molecular formula:
C₂₄H₂₅F
IUPAC Name:
4-(4-butylphenyl)-4'-ethyl-3-fluoro-1,1'-biphenyl
Specific details on test material used for the study:

Purity: 99.92%

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Samples were taken (in duplicate) from each concentration during the Limit Test at 0, 24, 48, 72 and 96 h. On each occasion, one sample was analysed after certain pre-treatments; the remaining samples were retained in case further analysis would be required.

25.0 ml collected water samples were diluted to 50 ml with 90% acetonitrile solution (acetonitrile:water = 90:10 (V:V)), and filtrated with 0.22 µm millipore membrane (discarding the initial 10 ml filtrates), the the filtrates were analysed by UPLC-PDA.

Test solutions

Vehicle:
yes
Details on test solutions:
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
The test solution was prepared by directly adding appropriate amounts in dilution water and then facilitating its dispersion by ultrasonification for 30 min.

In the limit test, WAFs (water accommodated fractions) solutions with a nominal concentration of 100 mg/L was prepared by adding appropriate amount of the test substance to the test medium in glass aspirator bottles with appropriate size. The mixing aspirator bottle was stoppered and the aqueous test substance mixture was stirred for 24 hours on a magnetic stirplate and a teflon stirbar at room temperature . The vortex height was set at least 10% of the static liquid height. At the end of the 24h, stirring was stopped and the test substance was allowed to settle down. The mixture was allowed to settle for 1 hour at room temperature. Then the aqueous portion removed through the outlet of the aspirator stirring bottle was the WAFs solution of the test substance without further filtration to remove the remaining undissolved test substance.

VEHICLE:
Good quality tap water which had been dechlorinated for at least 24 hours was used. The total hardness of the test water was 122.2. mg (CaCO3)/L, oxygen concentration was about 98% of the air saturation value and pH was 7.55 at room temperature.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish
- Strain: Dania rerio
- Source: Guangzhou Hongfa Aquatic Animals Culture Company in China
- Length at study initiation: 2.86 cm (average length)
- Weight at study initiation: 0.298 g (average weight)
- Feeding during test: commercial fish diet during acclimatization

ACCLIMATION
- Acclimation period: 7 days
- 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:
122 to 125 mg (CaCO3)/L
Test temperature:
22.6 - 22.9°C
pH:
7.57 - 7.73
Dissolved oxygen:
88% - 91%
Salinity:
no data
Nominal and measured concentrations:
Nominal Concentration: 100 mg/L
Mean measured concentration: 1.03 mg/L
Details on test conditions:
TEST SYSTEM
- Type (delete if not applicable): open
- Aeration: no
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated drinking water
- Culture medium different from test medium: no
- Intervals of water quality measurement: daily

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


TEST CONCENTRATIONS
- Range finding study: Yes
- Test concentrations: 0, 1, 10 and 100 mg/L
Reference substance (positive control):
no
Remarks:
The test with the reference substance is performed at least once each batch of fish as a means of assuring that the lab test conditions are adequate and have not changed significantly.

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 (fish)
Remarks on result:
other: meas. conc. 1.03 mg/L
Details on results:
- 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
Results with reference substance (positive control):
- Results with reference substance valid? Yes (the 24h-LC50 of potassium dichromate should range 200 mg/L to 400 mg/L)
- LC50: 269 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
During the test period, all fish in the control groups and treated groups were alive and appeared normal. The results showed that under valid static test conditions, the 96 h-LC50 of the test material to fish (Zebrafish, Brachydanio rerio) is greater than the nominal concentration of 100 mg/L WAFs (measured concentration 1.03 mg/L), while the maximum concentration causing no mortality (96 h-LC0) is equal to the nominal concentration of 100 mg/L WAFs (measured concentration 1.03 mg/L).
Executive summary:

The test was performed according to OECD 203. During the test period, all fish in the control groups and treated groups were alive and appeared normal. The results showed that under valid static test conditions, the 96 h-LC50 of the test material to fish (Zebrafish, Brachydanio rerio) is greater than the nominal concentration of 100 mg/L WAFs (measured concentration 1.03 mg/L), while the maximum concentration causing no mortality (96 h-LC0) is equal to the nominal concentration of 100 mg/L WAFs (measured concentration 1.03 mg/L).