Ethyl acetoacetate

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:

2009-11-17 to 2009-11-19

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
no data

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The limit concentration and the control were analytically verified via LC-MS/MS after 0 hours (fresh media) and after 48 hours (old media).
- Sampling method: For the old media, separate replicates without test organisms were prepared and incubated under test conditions. Recoveries of the test item should be within ± 20 % of the nominal value throughout exposure.
- Sample storage conditions before analysis:All samples were stored at room temperature, if necessary.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution (100 mg/L test item were weighed out) was freshly prepared with dilution water prior to application. Dispersion treatment was agitation.
- Eluate: Dilution water
- Differential loading: 1000 mg/L
- Controls: Dilution water without test item tested under the same conditions as the test group.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): None

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Feeding during test: No feeding
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strengt max. 20 µExm-2 x s-1


ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 255 (mg CaCO3/L)

Test temperature:

22.0 °C

pH:

Water Quality Parameters at the Beginning of the Test (0 hours, Fresh Media / measured in one additional replicate of the limit concentration and the control):
- Nominal Test Item Concentration 100 mg/L: pH-Value = 7.72
- Control: pH-value = 7.79

Water Quality Parameters at the End of the Test (48 hours, Old Media / measured in all replicates):
- Nominal Test Item Concentration 100 mg/L: pH-Values = 7.56-7.61
- Control: pH-value = 7.51-7.62

Dissolved oxygen:

Water Quality Parameters at the Beginning of the Test (0 hours, Fresh Media / measured in one additional replicate of the limit concentration and the control):
- Nominal Test Item Concentration 100 mg/L: Dissolved O2-Concentration = 8.32 mg/L
- Control: Dissolved O2-Concentration = 8.10 mg/L

Water Quality Parameters at the End of the Test (48 hours, Old Media / measured in all replicates):
- Nominal Test Item Concentration 100 mg/L: Dissolved O2-Concentration = 7.90-8.04 mg/L
- Control: Dissolved O2-Concentration = 7.92-8.09 mg/L

Salinity:

Freshwater

Nominal and measured concentrations:

Nominal Test Item Concentrations of 100 mg/L corresponds to measured concentrations (mean value of 2 injections - dilution factor taken into account) of 90.7 mg/L (recovery rate 91%). and 101 mg/L (recovery rate 100%). In the control samples, the measured concentrations were below the limit of quantification of the analytical method.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
Type: closed with parafilm
Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
Aeration: No
- Renewal rate of test solution (frequency/flow rate): One application at test start, static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Acc. to Directive 92/69/EEC Method C.2, Annex 1
- Conductivity: Dilution water at test start: 640 µS/cm
- Culture medium different from test medium: Elendt M4 acc. to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L
- Intervals of water quality measurement: At test start and test end

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1

TEST CONCENTRATIONS
- Spacing factor for test concentrations: None, limit test
- Range finding study: Test concentrations: 10 - 100 mg/L

IMMOBILIZATION RATES OF THE PRELIMINARY RANGE-FINDING TEST [%] (n = 20, divided into 2 replicates with 10 daphnids each):
- At concentrations of 100 and 10 mg/L nominal as well as in the control samples, no immobility was noted.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: None
- Mortality of control: 0%
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.67 (CI 1.63 - 1.71) mg/L

Reported statistics and error estimates:

The effects of the limit concentration of 1000 mg/L were determined directly from the observation data.

Percentage of Daphnids Incapable of Swimming after 24 and 48 h of Exposure:

(n = 20, divided into 4 replicates with 5 daphnids each)

IMMOBILIZATION [%]
Nominal Test Item Concentration [mg/L]	24 h					48 h
	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
100	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

 

Validity criteria fulfilled:

yes

Conclusions:

At the limit concentration 100 mg/L (nominal) of the test item Ethyl acetoacetate, no effect was observed. The EC50 (48h) on Daphnia Magna was found to be > 100 mg/l.

Executive summary:

In the acute immobilization test to Daphnia magna (STRAUS) the effects of the limit concentration 100 mg/L of the test item EAA were determined according to OECD 202 (2004). The limit test was conducted 2009 under static conditions over a period of 48 hours with analytical monitoring. 20 test organisms were exposed to the limit concentration and control.

At the limit concentration 100 mg/L (nominal) of the test item Ethyl acetoacetate, no effect was observed.

Description of key information

In the acute immobilization test to Daphnia magna (STRAUS) the effects of the limit concentration 100 mg/L of the test item EAA were determined according to OECD 202 (2004). The limit test was conducted 2009 under static conditions over a period of 48 hours with analytical monitoring. 20 test organisms were exposed to the limit concentration and control. At the limit concentration 100 mg/L (nominal) of the test item Ethyl acetoacetate, no effect was observed.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L