Registration Dossier

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report Date:

Materials and methods

Test guideline
according to
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. certificate)

Test material

Test material form:
solid: crystalline

Sampling and analysis

Analytical monitoring:

Test solutions

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 Cyclopropyl-enolether-acetal has a very low solubility in aquatic solutions so its concentration cannot be determined by UHPLC from aquatic solutions directly. The concentration of Cyclopropyl-enolether-acetal was determined only in the saturated stock solution by a non-validated HPLC method using a previous liquid/liquid extraction. The other test concentrations of the test item were calculated from the measured stock solution concentration. At the end of the study the concentrations of the test item in test vessels were not measured.

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:
Water media type:
Limit test:
Total exposure duration:
48 h

Test conditions

Test temperature:
The temperature inside the thermostat at the test vessels was measured and registered continuously by Extech SD200 thermometer and datalogger. The test vessels were covered with plastic lids and were kept in thermostat at 21 ± 1 °C with a 16/8 hours light/dark cycle.
The pH and dissolved oxygen were measured at the beginning and at the end of the test. At the beginning of the test the pH and the dissolved oxygen were measured from the remaining test solutions of the concurrent test vessels at each dilution levels. At the end of the test the pH and the dissolved oxygen were measured in each test vessel.
Dissolved oxygen:
There was not any significant change in the pH and oxygen concentration during the test.
Nominal and measured concentrations:
The stock solution was prepared by measuring 200.12 mg of test item into 2 000 ml Elendt M4 growth medium. After 15 minutes ultrasonication and two hours stirring the suspension was filtered through 0.2 μm WhatmannTM ME24 filter. The concentration of the filtrate (stock solution) was 0.12182 mg/L.
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 measured and calculated concentrations were applied: 0.122; 0.081; 0.061; 0.041 and 0.031 mg/L.
Details on test conditions:
Test procedure: The test was maintained under static conditions for a period of 48 hours.
At the start of the study each test vessels contained 50 ml of test solutions and five young daphnids. In the test four replicates total of 20 animals at each test concentration level were investigated. The test vessels were covered with plastic lids and were kept in thermostat at 21 ± 1 °C with a 16/8 hours light/dark cycle.
Each control vessels containing five young daphnids in Elendt M4 medium were prepared in four replicates and kept under the same static conditions. The number of immobilised and abnormally behaved daphnids was recorded after 24 and 48h. A daphnia is considered to be immobile if it is not able to swim within 15 seconds after a gentle agitation of the test vessel. The daphnids were not fed during the test.

Results and discussion

Effect concentrations
48 h
Dose descriptor:
Remarks on result:
not determinable
Details on results:
According to the test results Cyclopropyl-enolether-acetal has no toxic effect on Daphnia immobilisation even at its saturated concentration (0.122 mg/L).

Applicant's summary and conclusion

Validity criteria fulfilled:
In this Daphnia sp., Acute Immobilisation Test for Cyclopropyl-enolether-acetal the results indicated that the test item is not hazardous to the aquatic environment [48 hr EC50 (for Crustacea) > saturated concentration of Cyclopropyl-enolether-acetal (0.122 mg/L)