3,5-bis(2,4-dimethylcyclohex-3-en-1-yl)polyheterocycle

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

30 September 2016 - 16 December 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Specific details on test material used for the study:

Sa 57

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Water samples were taken (at least in duplicate) from each concentration during the definitive test at 0, 24, 48, 72 and 96 h. 50 mL collected water samples were extracted twice with 25 mL hexane. The combined organic phases were dried with rotary evaporator, and then redissolved with 2.00 mL acetonitrile. The final solutions were analyzed by UPLC.

Test solutions

Vehicle:

no

Details on test solutions:

In the range-finding test, the test solution was prepared by directly adding appropriate amounts of test substance into dilution water and then facilitating its dispersion by stirring for 30 min. The 1.00 mg/L test solution was colourless and clear. The test solutions with concentration of 10.0 and 100 mg/L showed white turbidity.
In the limit test, a W AF (water accommodated fraction) with a nominal concentration of 110 mg/L was prepared by adding an 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 by Teflon stir bar using a stirring rate about 500 rpm at room temperature (about 23°C ~ 25°C) for 24 hours. At the end of the 24 h, 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 W AF of the test substance without further filtration to remove any remaining undissolved test substance.

Test organisms

Test organisms (species):

other: Gobiocypris rarus

Details on test organisms:

The test fish Rare minnow, Gobiocypris rarus (Batch No.: F20160830G), were bred by the test institute.
Fish to be used in the test were held for 7 days in water of the quality and temperature to be used in the test.
A photoperiod of 16 hours light, provided by overhead fluorescent tubes, and 8 hours dark was maintained. The water quality was same with the test water. The oxygen concentration was more than 80% of the air saturation value. And the temperature was controlled between 22.8°C and 23.2°C.
The fish were fed twice daily during the holding period on proprietary fish food. They were held without food for approximately 24 hours before being placed into the test vessels.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

166 mg (CaCO3)/L to 175 mg (CaCO3)/L

Test temperature:

23.0°C to 23 .1°C

pH:

7,3 - 7,84

Dissolved oxygen:

74% and 95% of the air saturation

Nominal and measured concentrations:

nominal: 110 mg/L WAF
measured: 2.71 mg/L
The analytical results showed that the concentration of the test substance was consistent in the test medium throughout the 96-hour test period (deviation within 20%). Thus a static procedure was reasonable.

Details on test conditions:

Static method was adopted in limit test. The concentration of 110 mg/L WAF was assigned in the limit test. One control group was also used in the test. Three replicates, each containing 3L of test solution, were assigned for each treatment group and control group, while the initial number of test fish was 7 for each replicate.
The test fish were randomly chosen and put in appropriate test solutions after the temperature had been adjusted to the required value. This was done within 30 min.
During the test, the following conditions were maintained:
-Light: 16 hours photoperiod daily;
-Temperature: 23.0°C to 23.1 °C;
-Oxygen concentration: 74% to 95% the air saturation; No aeration.
-Feeding: none.
At 24, 48, 72 and 96 h, the mortalities of the fish were recorded, and observations on individual behaviour were performed. Meanwhile, measurements of pH, dissolved oxygen and temperature were carried out and recorded daily.
Fish loading: 0.5 g to 0.6 g fish (wet weight) per litre of test medium.

Reference substance (positive control):

yes

Remarks:

Reference substance: Potassium dichromate, K2Cr201, CAS: 7778-50-9; Purity: 99.8%; Lot number: LVC0052; J&K Scientific Ltd.

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 110 mg/L

Nominal / measured:

nominal

Conc. based on:

other: water accommodated fraction (WAF)

Basis for effect:

mortality (fish)

Details on results:

During the test period, all fish in the control groups and treated groups were alive and appeared
normal.

Results with reference substance (positive control):

Reference substance: Potassium dichromate, K2Cr20 7, CAS: 7778-50-9; Purity: 99.8%; Lot number: L VC0052; J&K Scientific Ltd. The test with the reference substance is performed at least once with each batch of fish as a means of assuring that the laboratory test conditions are adequate and have not changed significantly. The resulting 24 h-LC50 should be in the range of200 mg/L to 400 mg/L. Otherwise, the test is regarded as invalid.
The recent results (Study No: S2016RI001-07) were LC50(24h) 325 mg/L (95% Confidence Limit 286~371 mg/L).

Reported statistics and error estimates:

Trimmed Spearman-Karber Method (Version 1.5, USEPA) can be used to calculate the LC50 and 95% confidence limits. Due to its low toxicity to fish in this study, only the LC50 and 95% confidence limits of the reference substance was calculated by this method.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The results show that under valid static test conditions, the 96 h-LC50 of Sa57 to fish (Rare minnow, Gobiocypris rarus) is greater than the nominal concentration of the 110 mg/L WAF, while the maximum concentration causing no mortality (96h-LC0) was equal to the nominal concentration of 110 mg/L WAF.

96 h-LC50 > 110 mg/L WAF;
96 h-LC0 = 110 mg/L WAF.

Executive summary:

This test was designed to determine the acute toxicity of test substance (Sa57) to fish (Gobiocypris rarus). The test fish were exposed for 96 hours to the test solution. The study comprised at least one range-finding test followed by a Limit Test to determine the 96-hour median lethal concentration (LC50) of the test substance.

During the test period, all fish in the control groups and treated groups were alive and appeared normal.

The results show that under valid static test conditions, the 96 h-LC50 of Sa57 to fish (Rare minnow, Gobiocypris rarus) is greater than the nominal concentration of the 110 mg/L WAF, while the maximum concentration causing no mortality (96h-LC0) was equal to the nominal concentration of 110 mg/L WAF.

 

96 h-LC50 > 110 mg/L WAF;

96 h-LC0 = 110 mg/L WAF.