(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 aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

4th November 2018 to 7th November 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation 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. 202 "Daphnia sp., Acute Immobilisation 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:

yes

Remarks:

acetonitrile

Details on test solutions:

Method of preparation of stock solution:
A stock solution of nominal concentration 2000 mg/L was prepared to dilute test concentrations. A weight of 0.20400 g test item was dissolved in acetonitrile to 100 mL

Method of preparation of test solutions:
The stock solution was used to prepare the working standard solutions with acetonitrile of 0.002, 0.004, 0.02, 0.04, 0.1, 0.2, and 0.4 mg/L

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia of stock culture were originally obtained from the College of Applied Chemistry of China Agricultural University The test Daphnia were derived from a same healthy pre-cultured population (i.e. showing no signs of stress such as high mortality, presence of males and ephippia, delay in the production of the first brood, discoloured animals, etc.). The pre-cultured animals were maintained in culture conditions (light, temperature, and medium) similar to those to be used in the test. The test animals were less than 24 hours old and not the first brood progeny. The batch used for this definitive test was Daphnia C20181105.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

24hr, 48 hr

Test conditions

Hardness:

160 mg/L (as CaCO3)

Test temperature:

18-22°C, for each test constant within ± 1 °C

pH:

6-9

Dissolved oxygen:

not less than 3 mg/L during the test period

Nominal and measured concentrations:

Based on a preliminary test, the test concentrations were used as follows:

Nominal amount of test item 1.0006g was weighed and made up to 10L with test water to give 100 mg/L test solution.
2.5, 1.25, 0.625 and 0.313 L of saturated solution was pipetted into 2.5, 3.75, 4.375 and 4.687 test water to get 50%, 25% 12.5% and 6.25% saturated concentrations.

Measured concentrations: the determined concentrations in fresh media were 1.67, 3.33, 6.67, 13.1 and 29.5 mg/L. The determined concentrations in old media were 1.54, 2.93, 5.95, 11.1 and 26.6 mg/L. The overall determined concentrations were 1.61, 3.13, 6.31, 12.1 and 28.1 mg/L.

Details on test conditions:

The test solution of 100 mg/L was prepared by dissolving the test item in test water by magnetic stirring for 24 hours, and other solutions were diluted from the test solution of 100 mg/L. The test animals were Daphnia magna and 5 animals were used for each group. The solutions were not changed. The details were as following: The overall determined concentrations were 1.61, 3.13, 6.31, 12.1 and 28.1 mg/L. Test solutions were added into 4 test vessels respectively, and each vessel contained about 100 mL of the solution.5 daphnids were placed into each vessel. The test solutions were not renewed during the test.
Observation intervals: 24 and 48 hours after exposure.

Each test vessel was checked for immobilised daphnids. In addition to immobility, any abnormal behaviour or appearance was recorded. Those animals that are not able to swim within 15 seconds, after gentle agitation of the test vessel were considered to be immobilised (even if they can still move their antennae).

Reference substance (positive control):

yes

Remarks:

Potassium Dichromate

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.5 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

other: Immobilisation

Reported statistics and error estimates:

Trimmed Spearman-Karber (TSK) method (EPA, VERSION 1.5)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

According to the results of test and based on the arithmetic mean concentrations of fresh and old solutions, the 48-hour ECs, of E Stage 3 intermediate to Daphnia magna was calculated using Trimmed Spearman-Karber (TSK) method (EPA, VERSION 1.5) to be 3.5mg/L, and 95% confidence limits were 2.93 ~ 4.17 mg/L

Executive summary:

An Acute Immobilisation study of E Stage 3 intermediate was conducted with Daphnia magna over a 48-hour exposure period. The definitive test was carried out with overall determined concentrations of 1.61, 3.13, 6.31, 12.1 and 28.1 mg/L. A blank control was performed concurrently. 4 replicates were used for the control and test concentrations with 5 daphnids for each replicate. The test was performed with a static water system and the test solutions were not renewed during the test.

The  concentration  of  test  item  was  determined  in  samples  of test  media  by HPLC-MS/MS method.   

According to the results of test and based on the arithmetic mean concentrations of fresh and old solutions, the 48-hour EC₅₀, of E Stage 3 intermediate to Daphnia magna was calculated by the Trimmed  Spearman-Karber (TSK) method (EPA, VERSION 1.5)  to be 3.5 mg/L, and 95% confidence limits were 2.93 ~ 4.17 mg/L.