Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes
Specific details on test material used for the study:
Identification: Vinyl 2-ethylhexanoate
CAS Number: 94-04-2
EC Number: 202-297-4
Batch: PM9CEH01
Purity: 99.6%
Physical state / Appearance: Clear colorless liquid
Expiry date: 04 January 2022
Storage conditions: Room temperature in the dark over silica gel
Analytical monitoring:
yes
Test organisms (species):
Daphnia magna
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Details on test conditions:
Experimental Design and Study Conduct
Preliminary Media Preparation Trial
Preliminary solubility work conducted indicated that the test item was practically insoluble in water using traditional methods of preparation e.g. ultrasonication and high shear mixing.
Based on this information the test item was categorized as being a ‘difficult substance’ as defined by the OECD Guidance Document on Aqueous-Phase Aquatic Toxicity Testing of Difficult Test Chemicals (OECD 2019). Therefore a media preparation trial was conducted in order to determine the solubility of the test item under test conditions (see Annex 4).

Range-finding Test
The results obtained from the preliminary media preparation trial conducted indicated that a dissolved test item concentration of approximately 14 mg/L could be obtained using a saturated solution (shake flask) method of preparation.
The test concentrations to be used in the definitive test were determined by a preliminary range-finding test.
Due to the potentially volatile nature of the test item, testing was conducted in completely filled, sealed test vessels (120 mL glass) in order to minimize possible losses due to volatilization.
In the range-finding test Daphnia magna were exposed to a series of nominal test concentrations of 0.1, 1.0, 10 and 100% v/v saturated solution.
Nominal amounts of test item (56.5 mg) were separately dispensed in duplicate under the surface of 565 mL of test water in completely filled test vessels which were sealed immediately. The flasks were shaken at approximately 150 rpm at a temperature of 24 °C for 24 hours. After 24 hours, the contents of the flasks were pooled prior to the removal of any undissolved test item by filtration through a 0.2 μm Gelman Acrocap filter (first approximate 100 mL discarded in order to pre-condition the filter) to give a 100% v/v saturated solution. This saturated solution was then further diluted to give further stock solutions of 0.10, 1.0 and 10% v/v saturated solution of the test item.
Each prepared concentration was inverted several times to ensure adequate mixing and homogeneity.
In the range-finding test five daphnids were placed in each test and control vessel and maintained in a temperature controlled room maintaining the water temperature at 18 to 22 °C with a maximum deviation of ±1 °C and a photoperiod of 16 hours light and 8 hours darkness for a period of 48 hours with 20 minute dawn and dusk transition periods. Two replicate test and control vessels were prepared. Each test and control vessel was completely filled and sealed (120 mL glass) in order to minimize possible losses due to volatilization. After 24 and 48 hours the number of immobilized daphnids were recorded.
The control group was maintained under identical conditions but not exposed to the test item.
A sample of each test concentration was taken for immediate chemical analysis at 0 hours from the bulk test preparation and 48 hours from the pooled replicates in order to determine the stability of the test item under test conditions. A duplicate set of samples was taken on each occasion and stored frozen for further analysis if required.

Definitive Test
Based on the results of the range-finding test the following test concentrations were assigned to the definitive test: 4.5, 10, 22, 48 and 100% v/v saturated solution.
Nominal amounts of test item (56.5 mg) were separately dispensed in quadruplicate under the surface of 565 mL of test water in completely filled test vessels which were sealed immediately. The flasks were shaken at approximately 150 rpm at a temperature of 24 °C for 24 hours. After 24 hours, the contents of the flasks were pooled prior to the removal of any undissolved test item by filtration through a 0.2 μm Gelman Acrocap filter (first approximate 100 mL discarded in order to pre-condition the filter) to give a 100% v/v saturated solution.
A series of dilutions were made from this saturated solution to give further test concentrations of 4.5, 10, 22 and 48% v/v saturated solution.
Each prepared concentration was inverted several times to ensure adequate mixing and homogeneity.
The concentration and stability of the test item in the test preparations was verified by chemical analysis at 0, 24 and 48 hours (see Annex 5).

Exposure Conditions
Due to the potentially volatile nature of the test item, testing was conducted in completely filled, stoppered test vessels (120 mL glass flasks) in order to minimize possible losses due to volatilization.
At the start of the test five daphnids were placed in each test and control vessel at random, in the test preparations. Four replicate test and control vessels were prepared. Each test and control vessel was completely filled and stoppered in order to minimize possible losses due to volatilization. and maintained in a temperature controlled room maintaining the water temperature at 18 to 22 °C with a maximum deviation of ±1 °C and a photoperiod of 16 hours light (between 200 and 1200 Lux) and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified, received no food during exposure and the test vessels were not aerated.
The control group was maintained under identical conditions but not exposed to the test item.
Semi-static test conditions were employed in the test in an effort to maintain dissolved test item concentrations. For the test media renewal at 24 hours, the test concentrations were freshly prepared and the daphnids transferred by wide bore pipette from the 24-Hour old test media into the fresh test media.
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
ca. 1.2 mg/L
95% CI:
1 - 1.5
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mobility
Validity criteria fulfilled:
yes
Conclusions:
Exposure of Daphnia magna to the test item gave the following results based on the geometric mean measured test concentration:
Time Point EC50 95% Confidence No Observed Effect Lowest Observed
(Hours) (mg/L) Limits (mg/L) Concentration (NOEC) Effect Concentration
(mg/L) (LOEC) (mg/L)
48 1.2 1.0 - 1.5 0.36 0.97
Executive summary:

Introduction
A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.


Methods
Preliminary solubility work conducted indicated that it was not possible to obtain a testable solution of the test item using traditional methods of preparation e.g. ultrasonication and high shear mixing.
A preliminary media preparation trial indicated that a dissolved test item concentration of approximately 13 mg/L was obtained from a saturated solution (shake flask) method of preparation indicating this to be the limit of water solubility of this item under test conditions.
Due to the potentially volatile nature of the test item, testing was conducted in completely filled and sealed test vessels in order to minimize possible losses due to volatilization.
Following a preliminary range-finding test, 20 daphnids (4 replicates of 5 animals) were exposed to an aqueous solution of the test item at nominal concentrations of 4.5, 10, 22, 48 and 100% v/v saturated solution for 48 hours at a temperature of approximately 20 °C to 21 °C under semi-static test conditions. The test item solutions were prepared by employing a shake flask method of preparation, this method of preparation was undertaken due to the lack of test item found during a traditional saturated solution approach (due to the potential volatility of the test item). The test item solutions were prepared in four replicates by shaking in excess (100 mg/L) of the test item in test water at approximately 150 rpm, at a temperature of 24 °C for 24 hours. After the shaking period the contents of the replicate flasks were pooled and any undissolved test item was removed by filtration through a 0.2 μm Gelman Acrocap filter (first approximate 100 mL was discarded in order to pre-condition the filter) to give a 100% v/v saturated solutions of the test item. This saturated solution was then further diluted to give the 48, 22, 10 and 4.5% v/v saturated solutions of the test item.
Immobilization and any adverse reactions to exposure were recorded after 24 and 48 hours.


Results
Analysis of the fresh test preparations at 0 and 24 hours showed measured test concentrations to range from 0.44 to 14 mg/L. Measured test concentrations of the aged test preparations at 24 and 48 hours showed measured test concentrations to range from 0.27 to 12 mg/L. As not all test concentrations were maintained within ± 20% of the measured initial test concentration it was considered appropriate to calculate the results based on the geometric mean measured test concentration in order to give a “worst case” analysis of the data. The geometric mean measured test concentrations were determined to be 0.36, 0.97, 2.3, 5.3 and 12 mg/L.
Exposure of Daphnia magna to the test item gave the following results based on the geometric mean measured test concentration:

Description of key information

The EC50 for freshwater daphnia data is 1.2 mg/L. 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Effect concentration:
1.2 mg/L

Additional information