2-Propenoic acid, 2-[2-(ethenyloxy)ethoxy]ethyl ester

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between May 12th, 2006 and July 31st, 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of inspection: April 2005 Date of Signature: November 2005

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
The following concentrations of 2-(2-Vinyloxyethoxy) ethyl acrylate were tested: nominal 25, 50 and 100 mg/L. Additionally, a biological control was tested in parallel (test water without test item).

- Sampling method:
For the determination of the actual test item concentrations, the following samples were taken:
Just before the start of the test:
- duplicate samples from each test medium (without daphnids)
- duplicate samples from the control (without daphnids)
After 24 and 48 hours:
(stability samples) - duplicate samples from each test medium
- duplicate samples from the control:
For the 24-hour and 48-hour stability samples additional flasks with adequate volumes of the freshly prepared test media of all test concentrations and the control were incubated during the test period under the same conditions as in the actual test (but without daphnids).

- Sample storage conditions before analysis:
tested immediately after sampling

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)

The test medium of the highest nominal concentration of 100 mg/L was prepared by dissolving 60.9 mg of test item completely in 600 mL of test water using ultrasonic treatment for 10 minutes and intense stirring for 10 minutes at room temperature. Adequate volumes of this test medium were diluted with test water to prepare the test media with the lower test item concentrations. The test media were prepared just before introduction of the daphnids (i.e., start of the test).

- Eluate:
Not applicable

- Controls:

For evaluation of the quality of the daphnia clone and the experimental conditions, potassium dichromate is tested as a positive control at least once a year. The latest result of the positive control test in September 2005 (48-hour EC50: 0.81 mg/L, RCC Study No. A28102) showed that the toxic performance was valid and within the historical range of the RCC laboratory (from 1996 to 2005: 48-hour EC50: 0.53-1.1 mg/L).

- Chemical name of vehicle :
Not applicable

- Concentration of vehicle in test medium:
Not applicable

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name:
Water Flea

- Source:
originally supplied by the University of Sheffield/UK in 1992 bred at RCC since then.

- Age at study initiation:
Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were 6 to 24 hours old.

- Feeding during test:
Received no food during exposure

ACCLIMATION
- Acclimation period:
Not stated

- Acclimation conditions:
No acclimitisation reported.

- Type and amount of food:
Each culture was fed daily with a suspension of algae (Chlorella sp.).
The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

- Health during acclimation:
Not applicable

Test Water:
The reconstituted water used for both the range-finding and definitive tests was the same as that used to maintain the stock animals.


Reconstituted Water
i) Stock Solutions
a) CaCl2.2H2O 2.0 mmol/L (= 294 mg/L)
b) MgSO4.7H2O 0.5 mmol/L (= 123 mg/L)
c) NaHCO3 0.75 mmol/L (= 65 mg/L)
d) KCl 0.075 mmol/L (= 5.8 mg/L)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Observations were taken at 24 and 48 hours

Hardness:

The reconstituted water had an approximate theoretical total hardness of 2.5 mmol/L (= 250 mg/L as CaCO3)

Test temperature:

At the start and at the end of the test, the water temperature were determined in each test concentration and the control.
Water temperature: 19 - 20°C during the test period (Table 3). The test was performed in a temperature-controlled room (room temperature continuously monitored).

pH:

The pH was measured at the start of the test, after 24 hours and at the end of the test. The pH values were calculated in the range of 7.8 to 8.0.

Dissolved oxygen:

At the start and at the end of the test, the dissolved oxygen concentration was determined in each test concentration and the control.
At the beginning, after 24 hours, and the end of the test period, the dissolved oxygen concentrations in the test media and the control were at least 8.8 +/- 0.2 O2 mg/L,

Salinity:

freshwater used.

Nominal and measured concentrations:

The nominal test item concentrations tested were 6.3, 12.5, 25, 50 and 100 mg/L. Additionally, a control was tested in parallel.

Details on test conditions:

TEST SYSTEM
- Test vessel:
The test was performed in 100 mL glass beakers filled with 50 mL of test medium. The beakers were covered with glass plates to reduce the loss of water and to avoid the entry of dust into the solutions. The test vessels were labeled with the RCC study number and all necessary additional information to ensure unmistakable identification.

- Type (delete if not applicable):
closed

- Aeration:
The test water was aerated prior to the start of the study until oxygen saturation was reached. During the test period, the test water was not aerated.

- Type of flow-through (e.g. peristaltic or proportional diluter):
None.

- Renewal rate of test solution (frequency/flow rate):
None.

- No. of organisms per vessel:
not applicable

- No. of vessels per concentration (replicates):
Four test vessels were used for each test and control group.

- No. of vessels per control (replicates):
At each test concentration and for the control, 20 daphnids were used divided into four replicates of five daphnids each.

- No. of vessels per vehicle control (replicates):
Duplicate test vessels were used for each test and control group.

- Biomass loading rate:
Not recorded.

- Source/preparation of dilution water:
Reconstituted Water
i) Stock Solutions
a) CaCl2.2H2O 2.0 mmol/L (= 294 mg/L)
b) MgSO4.7H2O 0.5 mmol/L (= 123 mg/L)
c) NaHCO3 0.75 mmol/L (= 65 mg/L)
d) KCl 0.075 mmol/L (= 5.8 mg/L)

Water Hardness : 2.5 mmol/L (= 250 mg/L as CaCO3)
Alkalinity : 0.8 mmol/L

- Alkalinity:
Alkalinity : 0.8 mmol/L

- Ca/mg ratio:
Ratio of Ca : Mg = 4 : 1 (based on molarity)

Na : K = 10 : 1 (based on molarity)

- Conductivity:
Not recorded

- Culture medium different from test medium:
The reconstituted water used for both the range-finding and definitive tests was the same as that used to maintain the stock animals.

- Intervals of water quality measurement:
Not recorded

OTHER TEST CONDITIONS

- Photoperiod:
A 16-hour light to 8-hour dark photoperiod (with a 30 minute transition period).

- Light intensity:
light period was between approximately 470 and 640 Lux.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Any immobilisation or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure.

TEST CONCENTRATIONS:

- Spacing factor for test concentrations:
The following concentrations of 2-(2-Vinyloxyethoxy) ethyl acrylate were tested: nominal 6.3, 12.5, 25, 50 and 100 mg/L.

- Justification for using less concentrations than requested by guideline:
Not applicable

- Range finding study:

A pre experiment using a non GLP method was undertaken.

A static test without test medium renewal was chosen since in a pre-experiment (without GLP) the test item concentration in the test medium was constant during the test period of 48 hours.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

96 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

55 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL 40 - 77mg/L

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: None stated
- Observations on body length and weight: Not stated in report
- Other biological observations: None stated
- Mortality of control: None
- Other adverse effects control: None
- Abnormal responses: none stated
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none reported

The biological results are listed in Table 1.

During the first 24 hours of the test no immobility of the test organisms was observed in the control and up to and including the test item concentration of 50 mg/L. At the highest test concentration of 100 mg/L, 55% of the test organisms were found to be immobile.

The 24-hour EC50 of the test item was calculated to be 96 mg/L (95% confidence limits not calculable). The 24-hour EC0 was 50 mg/L, the 24-hour EC100 was > 100 mg/L.

After 48 hours of exposure no immobility of the test organisms was observed in the control and up to and including the test item concentration of 25 mg/L. At the concentration of 50 mg/L, 35% of the daphnids were found to be immobile. At the highest test concentration of 100 mg/L all test organisms were immobile after the exposure period of 48 hours. The 48-hour EC50 was calculated to be 55 mg/L with 95% confidence limits from 40 to 77 mg/L.

The 48-hour EC0 and the 48-hour NOEC (highest concentration tested without toxic effects after 48 hours) of 2-(2-Vinyloxyethoxy) ethyl acrylate were both 25 mg/L, since no immobilization was observed in the test organisms exposed to the test item at levels of up to and including this test concentration. The 48-hour EC100 was 100 mg/L.

No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions throughout the entire test duration.

Results with reference substance (positive control):

- Results with reference substance valid?
Yes

- Mortality:
No mortalities recorded.

- EC50/LC50:
After 48 hours of exposure no immobility of the test organisms was observed in the control.

Reported statistics and error estimates:

None stated in report

Table 1:

Nominal test item concentration	No. of daphnids tested	Immobilized daphnids after 24 hours		Immobilized after 48 hours
		No.	%	No.	%
control	20	0	0	0	0
6.3	20	0	0	0	0
12.5	20	0	0	0	0
25	20	0	0	0	0
50	20	0	0	7	35
100	20	11	55	20	100

Validity criteria fulfilled:

yes

Conclusions:

After 48 hours of exposure no immobility of the test organisms was observed in the control and up to and including the test item concentration of 25 mg/L. At the concentration of 50 mg/L, 35% of the daphnids were found to be immobile. At the highest test concentration of 100 mg/L all test organisms were immobile after the exposure period of 48 hours. The test gave an EC50 value of 55mg/l and an NOEC value of 25 mg/l

Executive summary:

The acute toxicity of the test item 2-(2-Vinyloxyethoxy) ethyl acrylate to Daphnia magna was determined in a 48-hour static test according to the EU Commission Directive 92/69/EEC, Part C.2 (1992), and the OECD Guideline for Testing of Chemicals, No. 202 (2004). The nominal test item concentrations tested were 6.3, 12.5, 25, 50 and 100 mg/L. Additionally, a control was tested in parallel. The analytically determined concentrations of the test item in the analyzed test media from the sampling times at the start, after 24 hours and at the end of the test were between 93 and 101% of the nominal values. Thus, the test item 2-(2-Vinyloxyethoxy) ethyl acrylate was stable during the test period of 48 hours under the conditions of the test, and the reported biological results are based on the nominal concentrations of the test item.

The biological test results:

− 24-hour EC50: 96 mg/L (95% confidence limits not calculable)

− 24-hour EC0: 50 mg/L

− 24-hour EC100: > 100 mg/L

− 48-hour EC50: 55 mg/L (95% confidence limits: 40 - 77 mg/L)

− 48-hour EC0 and

48-hour NOEC: 25 mg/L

− 48-hour EC100: 100 mg/L

Description of key information

The 48-hour EC50 was calculated to be 55 mg/L The 48-hour EC0 and the 48-hour NOEC of 2-(2-Vinyloxyethoxy) ethyl acrylate were both 25 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

55 mg/L

Additional information

The acute toxicity of the test item 2-(2-Vinyloxyethoxy) ethyl acrylate to Daphnia magna was determined in a 48-hour static test according to the EU Commission Directive 92/69/EEC, Part C.2 (1992), and the OECD Guideline for Testing of Chemicals, No. 202 (2004).

The nominal test item concentrations tested were 6.3, 12.5, 25, 50 and 100 mg/L. Additionally, a control was tested in parallel. The analytically determined concentrations of the test item in the analyzed test media from the sampling times at the start, after 24 hours and at the end of the test were between 93 and 101% of the nominal values. Thus, the test item 2-(2-Vinyloxyethoxy) ethyl acrylate was stable during the test period of 48 hours under the conditions of the test, and the reported biological results are based on the nominal concentrations of the test item.

The biological test results:

− 24-hour EC50: 96 mg/L (95% confidence limits not calculable)

− 24-hour EC0: 50 mg/L

− 24-hour EC100: > 100 mg/L

− 48-hour EC50: 55 mg/L (95% confidence limits: 40 - 77 mg/L)

− 48-hour EC0 and 48-hour NOEC: 25 mg/L

− 48-hour EC100: 100 mg/L