trans-1-[4-(4-Propylcyclohexyl)-1-cyclohexen-1-yl]-4- (trifluoromethyl)-benzene

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Dec 12, 2015 - Mar 11, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

no OECD GLP
The study was conducted for a non-EU country.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Remarks:

Guidelines of Good Laboratory Practices on Chemical Testing (HJIT 155-2004, MEP, China)

Analytical monitoring:

yes

Details on sampling:

The concentrations of the test solutions were measured at both the start and the end of the exposureas weil as before and after each renewal. Samples of 20 mL were taken from treatment groups directly.They were injected to analyze concentration after the test solution samples were added 20 mL Hexane, extracted and taken Hexane phase to the sample bottles.

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Preparation: Prepare a solution/medium of nominal 100 mg/L and filter it to remove undissolved test material.
The test substance was weighed and dissolved into the test water with a constant volume of 5 L with 4
times to prepare the turbid liquid of 100 mg/L (nominal concentration) during the 96h test period (see
the table from below). The mixture solution was stirred for about 48 hours followed by filtrating with
a 0.45 μm membrane filter to prepare the test solution for the treated group. 5 L water were added into
the test vessel of the blank group.

- Preparation frequency: Every 24 hours.
- Storage: Test solutions were prepared just before use.

Test organisms (species):

other: Rare minnow (Gobiocypris rarus)

Details on test organisms:

TEST ORGANISM
Species: Rare minnow (Gobiocypris rarus)
Supplier: Wuchang district of wuhan city Jiahaibao fish shop
Generation No: FRA000-20151107-01
Sensitivity: LC50(24 h) of potassium dichromate was 345.635 mg/L
Terminal Body Length: 2.803 - 2.916 cm


ACCLIMATION
The fish were taken from the flowing water breeding system (which is part of the testing facility)
followed by a 7-day acclimatization period in the semi-static test system.
During the acclimatization period, light was set as 14 hours photoperiod daily. Temperature, pH
and dissolved oxygen concentration of the water were measured and recorded daily.
The fish were fed daily until 24 hours before the test started. Fish were not
selected for testing were returned to the fish breeding system.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

75 mg/L (CaCO3)

Test temperature:

23.6-24.4 °C

pH:

7.40 - 7.62

Dissolved oxygen:

81.5 - 99.7 %

Nominal and measured concentrations:

During the test, the dissolved test material concentration analytically were determined in the freshly prepared solution/medium (t=0h, 24h, 48 h, 72 h) and in the aged medium/solution before renewal (t=24h*, 48h*, 72h*, 96h*). The stability of the test material was determined by dividing the concentration t=24h by the concentration t=0h.
The ratio of the concentrations before renewal comparing with the concentration of the initial test solution were 60%~300. Therefore, the geometric average of the measured concentrations was used in this study.

Details on test conditions:

TEST SYSTEM
- 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: 0.641 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water

OTHER TEST CONDITIONS
- Photoperiod: 14 h light/10 h dark


EFFECT PARAMETERS MEASURED:
mortality, inactivity, abnormal swimming pattern, other abnormal behaviour recorded at 3, 6, 24, 48, 72 and 96 hours

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 100 mg/L (nominal)

Reference substance (positive control):

yes

Remarks:

potassium dichromate

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)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.004 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Results with reference substance (positive control):

LC50(24 h) of potassium dichromate was 345.635 mg/L

Validity criteria fulfilled:

yes

Conclusions:

There was no acutely toxic effect at maximum dissolved concentration of test substance, and the LC50 of 96 h of test substance on Gobiocypris rarus was greater than 0.004 mg/L based on the geometric average of the measured concentrations under the test conditions in this study.

Executive summary:

To evaluate the short-term toxicity effects of the test substance on Gobiocypris rarus with the determination of a 96hr LC50 in a semi-static aquatic system.

According to the result of the preliminary test, one treatment group of saturated solution (with the nominal concentration of 100 mg/L which was filtrated to remove undissolved test material by filter film of superthin glass fiber with 0.45 μm pore size) and one blank group (fish breeding water) were designed in the definitive test. Each group contained ten fish without replicates. The duration of the test was 96 hours. The test fish were exposed in closed bottles filled with the test solution under the semi-static condition. The test solution was renewed every 24 hours. Testing was performed under the condition of 14 hours photoperiod daily, 23.6~24.4 °C water temperature, pH of 7.40~ 7.62, 81.5%~99.7% air saturation value of dissolved oxygen concentration and without feeding. The symptom of fish were observed at 3, 6, 24, 48, 72 and 96 hours after the beginning of the exposure. The concentrations of the test substance were measured by GC-MS analysis at both the statt and the end of the exposure as weil as before and after each renewal.

At the end of the test, the cumulative mortalities were 0% and visible abnormalities were not found in either the blank group or the treated group. Initial analytically determined concentrations ranged from 0.001 to 0.013 mg/L, concentrations in the aged media ranged from 0.003 to 0.009 mg/L. The LC50 values were estimated based on the geometric average of the measured concentrations. The geometric average of the measured concentrations during test period of 96 hours was 0.004 mg/L in the treated group. The LC50(96h) value of the test material on Gobiocypris rarus was greater than 0.004 mg/L based on the measured concentrations (geometric average).

There was no acutely toxic effect at maximum dissolved concentration of test substance, and the LC50 of 96 h of test substance on Gobiocypris rarus was greater than 0.004 mg/L based on the geometric average of the measured concentrations under the test conditions in this study.

Description of key information

OECD 203: EC50 (96 h) > 100 mg/L (nominal) corresponding to 0.004 mg/L (measured geometric mean)