(1R)-2-[5-(2-halo-3-methoxyphenyl)-3-[2-halo-6-(trihalomethyl)benzyl]- 4-alkyl-2,6-dioxoheterocyclyl]-1-phenylethanaminium salicylate

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:

4th November 2018 to 9th November 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: MEP, P.R. China, the Guidelines for the Testing of Chemicals, Effects on Biotic Systems (the Second Edition), No. 203 "Fish Acute Toxicity Test". (2013.8)

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: EC, DG Health and Consumer Protection: Residues: Guidance for generating and reporting methods of analysis in support of pre-registration data reqs for Annex II (part A sec 4) & Annex III (Part A sec 5) of Dir 91/414 SANCO/3029/99 rev 4 11/07/00

GLP compliance:

yes

Test material

Test material information

Sampling and analysis

Analytical monitoring:

yes

Remarks:

HPLC-MS/MS

Details on sampling:

Analysis frequency: in 0-hour fresh solution and 48-hour old solution

Test solutions

Vehicle:

no

Details on test solutions:

A saturated solution (SS) at nominal concentration of 100 mg/L was prepared. A weight of 1.0006 and 1.0005g of test item was added to 10L water using a stirring rate of 460-490 rpm for 24 hours. Solutions were filtrated with 0.45 μm membrane.

2.5, 1.5, 0.625 and 0.313mL saturated solutions were pipetted and diluted respectively with 2.5, 3.75, 4.375 and 4.687mL test water to get 2.5, 12.5 and 6.25 % saturated concentration

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Zebra fish (Danio rerio) were propagated in the facility. No sign of disease or large mortalities
Acclimatation: Fish held under test conditions
Temperature: 23 ± 2°C
Light: 12 hours light / 12 hours darkness
Feeding: Brine shrimp twice daily on weekdays and once at weekend until 24 hours before test start
Body length: 2.1 - 2.9cm
Weight: 0.1808 - 0.3103g

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

3, 24, 48, 72 and 96 hours

Test conditions

Hardness:

10 to 250 mg/L (as CaCO3)

Test temperature:

23 ± 2°C

pH:

6.0 to 8.5

Dissolved oxygen:

Not less than 60% of air saturation value (ASV)

Nominal and measured concentrations:

Measured concentrations: the mean concentrations in fresh media were 1.66, 3.09, 6.37, 13.1 and 29.5 mg/L. The mean concentrations in old media were 1.38, 2.71, 5.63, 11.1 and
26.0mg/L. The overall mean concentrations were 1.52, 2.90, 6.00, 12.1 and 27.8mg/L.

Details on test conditions:

A saturated solution (SS) at nominal concentration of 100 mg/L was prepared. A weight of 1.0006 and 1.0005g of test item was added to 10L water using a stirring rate of 460-490 rpm for 24 hours. Solutions were filtrated with 0.45 μm membrane. 2.5, 1.5, 0.625 and 0.313mL saturated solutions were pipetted and diluted respectively with 2.5, 3.75, 4.375 and 4.687mL test water to get 2.5, 12.5 and 6.25 % saturated concentration

Test solutions were replaced every 48 hours, After the test solution was changed, the fish were transferred to freshly prepared solutions. A concurrent control group was conducted in parallel.
Control: 7 fish in dilution water (without test item) were tested under the same test conditions as test treatment

Observations were made 3, 24, 48, 72 and 96 hours after exposure. Fish are considered dead if there is no visible movement and if touching the caudal peduncle produces no reaction.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.43 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 96 hour LC50 of E stage 3 intermediate to Zebra fish was 3.43 mg/L, 95% confidence limits 2.7 - 4.33 mg/L

Executive summary:

A fish acute toxicity study of  E stage 3 intermediate was conducted with Zebra fish (Danio rerio) over a 96 h exposure period. 

Based on the results of the preliminary test, the definitive test was carried out at a saturated concentration of 100 mg/L, 50% saturated concentration, 25% saturated concentration, 12.5%  saturated concentration and 6.25% saturated concentration. The actual concentration of test solutions was measured by HPLC/MS/MS method. A concurrent blank control was performed in parallel. 7 fish were used for the test and control group. 

Based on the results of the preliminary test, a semi-static, renewed at 48 h with solution was used in the definitive test. Dissolved oxygen, pH and temperature were measured daily during the 96 h exposure period in the control and test solution. Temperature (in the water bath) was measured continuously by an automatic monitoring sytem.

During the 96 h exposure period no death or visible toxicity signs were observed in the control group and test group of 6.25%  saturated concentration. 2 fish died in test group 12.5% saturated concentration and 7 fish died in other test groups. 

The concentrations of the test system were determined in samples of test media by HPLC/MS/MS method. The measured concentrations were as follows:

The mean concentrations in fresh media were 1.66, 3.09, 6.37, 13.1 and 29.5 mg/L.

The mean concentrations in old media were 1.38, 2.71, 5.63, 11.1 and 26.0mg/L.

The overall mean concentrations were 1.52, 2.90, 6.00, 12.1 and 27.8mg/L.

Based on the results, the 96 hour LC₅₀ of E stage 3 intermediate to Zebra fish was 3.43 mg/L, 95% confidence limits 2.7 - 4.33 mg/L