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

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17th December 2018 to 8th January 2019

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 211 (Daphnia magna Reproduction Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: China Environmental Press, Guidelines for the Testing of Chemicals no. 211 Daphnia Reproduction Test (version 2. 2013)

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: EC, DG Health & Consumer Protection Residues Guidance for generation and reporting methods of analysis in support of pre-registration data requirements for Annex II and Annex III of 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 10 mg/L was prepared. A weight of 0.02g of test item was added to 2L water using a stirring rate of 450-470 rpm for 22-24 hours. Solutions were filtrated with 0.45 μm membrane.

150, 75, 37.5, 18.8 and 9.3mL saturated solutions were pipetted and diluted respectively with 1350, 1425, 1463, 1481 and 1491 mL test water to get 10, 5, 2.5, 1.25, and 0.625 % saturated concentration

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 C20181218.

Study design

Test type:

semi-static

Water media type:

freshwater

Total exposure duration:

21 d

Post exposure observation period:

Daily from Day 7

Test conditions

Hardness:

> 140 mg/L (as CaCO3)

Test temperature:

20 ± 2°C

pH:

6.0 to 9.0

Dissolved oxygen:

>3 mg/L

Nominal and measured concentrations:

Measured concentrations: the arithmetic mean concentrations in fresh media were 0.0533, 0.113, 0.226, 0.455 and 0.876 mg/L. The arithmetic mean concentrations in old media were 0.0550, 0.108, 0.219, 0.472 and 0.877 mg/L. The overall arithmetic mean concentrations were 0.0542, 0.111, 0.223, 0.461 and 0.877 mg/L.

Details on test conditions:

A saturated solution (SS) at nominal concentration of 10 mg/L was prepared. A weight of 0.02g of test item was added to 2L water using a stirring rate of 450-470 rpm for 22-24 hours. Solutions were filtrated with 0.45 μm membrane. 150, 75, 37.5, 18.8 and 9.3mL saturated solutions were pipetted and diluted respectively with 1350, 1425, 1463, 1481 and 1491 mL test water to get 10, 5, 2.5, 1.25, and 0.625 % saturated concentration.

Test solutions were replaced every 48 hours. After the test solution was changed, the daphnia were transferred to different chambers with freshly prepared solutions. A concurrent control group was conducted in parallel.
Control: 10 daphnids in dilution water (without test item) were tested under the same test conditions as test treatment.

The offspring produced by each parent were removed and counted daily. Mortality among the parent animals were recorded daily. The reproductive outputs per surviving parent animals were recorded.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.111 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality: Number of living offspring produced per surviving parental animal {for Daphnia magna, TG 211}

Effect concentrations 2

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.223 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality: Number of living offspring produced per surviving parental animal {for Daphnia magna, TG 211}

Effect concentrations 3

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.877 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Body Length

Effect concentrations 4

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.233 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: First brood time

Effect concentrations 5

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

0.464 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: First brood time

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The total number of juveniles per parent, body length and first brood time were analysed by a 2-sided T-test using a 5% significance level to obtain the No Observed Effect Concentration (NOEC) and the Lowest Observed Effect Concentration (LOEC)
The 21d NOEC and LOEC for each parameter are as follows:
Total no of juveniles per parent: NOEC 0.111 mg/L LOEC 0.223 mg/L
Body length: NOEC 0.877 mg/L
First brood time: NOEC 0.233 mg/L LOEC 0.464 mg/L

Executive summary:

A reproduction test with E Stage 3 intermediate was conducted with Daphnia magna over a 21-day exposure period. The definitive test was conducted with five concentrations of with arithmetic mean concentrations of 0.0542, 0.111, 0.223, 0.461 and 0.877 mg/L., concurrently with a blank control group. 10 animals were individually held at each test concentration and control. The semi-static test was performed with renewal of the test solutions every 48 hours.

During the test, no deaths or visible toxic signs were observed in control group and lower test groups; 4 deaths were observed in the 10% SS. There is no difference in the total no of juveniles between 0.625, 1.25 and 2.5 % SS and the control. There is significant difference between 5 and 10% SS and the control. There is no difference in body length between the test groups and the control group. There is no difference in the first brood time between 0.625, 1.25 and 2.5 % SS and the control. There is significant difference between 5 and 10% SS and the control. 

Measured concentrations: the arithmetic mean concentrations in fresh media were 0.0533, 0.113, 0.226, 0.455 and 0.876 mg/L. The arithmetic mean concentrations in old media were 0.0550, 0.108, 0.219, 0.472 and 0.877 mg/L. The overall arithmetic mean concentrations were 0.0542, 0.111, 0.223, 0.461 and 0.877 mg/L.

The total number of juveniles per parent, body length and first brood time were analysed by a 2-sided T-test using a 5% significance level to obtain the No Observed Effect Concentration (NOEC) and the Lowest Observed Effect Concentration (LOEC)
The 21d NOEC and LOEC for each parameter are as follows:
Total no of juveniles per parent: NOEC 0.111 mg/L LOEC 0.223 mg/L
Body length: NOEC 0.877 mg/L
First brood time: NOEC 0.233 mg/L LOEC 0.464 mg/L