3-ethyl 5-methyl 4-(2-chlorophenyl)-1,4-dihydro-2-[2-(1,3-dihydro-1,3-dioxo-(2H)isoindol-2-yl)-ethoxymethyl]-6-methyl-3,5-pyridinedicarboxylate

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:

48 h

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

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-ethyl 5-methyl 4-(2-chlorophenyl)-1,4-dihydro-2-[2-(1,3-dihydro-1,3-dioxo-(2H)isoindol-2-yl)-ethoxymethyl]-6-methyl-3,5-pyridinedicarboxylate

EC Number:

413-410-3

EC Name:

3-ethyl 5-methyl 4-(2-chlorophenyl)-1,4-dihydro-2-[2-(1,3-dihydro-1,3-dioxo-(2H)isoindol-2-yl)-ethoxymethyl]-6-methyl-3,5-pyridinedicarboxylate

Cas Number:

88150-62-3

Molecular formula:

C28H27ClN2O7

IUPAC Name:

3-ethyl 5-methyl 4-(2-chlorophenyl)-2-{[2-(1,3-dioxo-2,3-dihydro-1H-isoindol-2-yl)ethoxy]methyl}-6-methyl-1,4-dihydropyridine-3,5-dicarboxylate

Test material form:

solid: crystalline

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

yes

Details on test solutions:

The test organism was bred and held in Elendt M4 medium which was also used for preparing of test item solutions. The stock solution was prepared by measuring 200.31 mg of test item into 2 000 ml Elendt M4 growth medium. After one hour stirring and ultrasonication the suspension was filtered through 0.2 μm WhatmannTM ME24 filter. The concentration of the filtrate (stock solution) was 0.01378 mg/L.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Test species: Daphnia magna Straus
Age: < 24 hours neonates
Origin: National Institute of Environmental Health
Date of arriving: 16 May 2013
Culture conditions: The test species is maintained in Elendt M4 medium with culture conditions at 21±1 °C and a light/dark cycle of 16/8 hours with light intensities of less than 1000 Lux. The cultures are fed with centrifugalized green alga (Pseudokirchneriella subcapitata) cells daily at weekdays.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

At each concentration level four replicates were prepared. Each test vessel contained 5 daphnids in 50 ml test solution. The test vessels were covered with plastic lids and were kept in thermostat at 21 ± 1 °C and 16/8 hours light/dark cycle. The controls were kept under the same conditions in Elendt M4 medium.

Nominal and measured concentrations:

The toxic property of test item was investigated at five concentrations which were prepared by different dilutions of stock solution of test item. Elendt M4 medium was used for dilution of stock solution. In the test the following calculated concentrations were applied: 0.0138; 0.0092; 0.0069; 0.0046 and 0.0034 mg/L.

Details on test conditions:

Selection of young daphnids: On the first day of the test, all of juvenile daphnids were removed from the selected breeding cultures by sieving. Juvenile daphnids were disposed. Next day the neonates (which are younger than 24 hours) were separated from the parental culture by filtering and were transferred into test vessels.
Preparation of stock solution: The preparation of stock and test solutions was based on the preliminary non GLP range finding test [9]. As the Range Finding Test was not performed in compliance with the GLP-Regulations it is excluded from the Statement of Compliance in the final report, but the raw data of this test are archived under the study code of present study. For the stock solution 200.31 mg of test item was measured into 2 000 ml Elendt M4 medium. After one hour stirring and ultrasonication the suspension was filtered through 0.2 μm WhatmannTM ME24 filter. The concentration of the filtrate (stock solution) was 0.01378 mg/L.
Preparation of test solutions: Five dilution levels were prepared by mixing appropriate volume of Elendt M4 medium and stock solution of the test item. The maximum separation factor between of test vessels was 1.50. The following table shows the preparation of the different dilution levels. The volume of the prepared test solutions was 300 ml.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

48 h

Remarks on result:

not determinable because of methodological limitations

Details on results:

The evaluation of the toxic effects of the test item was based on the following calculated concentrations 0.0138; 0.0092; 0.0069; 0.0046 and 0.0034 mg/L.
The effective concentrations (ECx) were calculated with Probit analysis using linear max. Likelihood regression. The EC10, EC20 and EC50 were calculated for immobility at both 24 h and 48 h period. As the Amlodipine Phthaloyl has no toxic effect on Daphnia immobilisation even at its saturated concentration (0.0138 mg/L) the values of NOEC and LOEC were not calculated.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Amlodipine Phthaloyl has no toxic effect on Daphnia immobilisation even at its saturated concentration.
In this Daphnia sp., Acute Immobilisation Test for Amlodipine Phthaloyl the results indicated that the test item is not hazardous to the aquatic environment