Ethylene di(acetate)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 Jul - 21 Jul 2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control, 7.5, 15, 30, 60, 120 mg/L
- Sampling method: Samples of 1.4 mL were taken from mid-depth of all replicates and submitted for concentration verification at time 0-, 48- (seasoned and renewed), and 96-hours. Samples were submitted for analysis at time 24-hours due to complete mortality in both test vessels containing nominally 120 mg/L. The samples were combined with a nominal 0.14 mL of internal standard.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A stock solution of nominal 24 mg/mL was prepared in dilution water. Appropriate volume of stock solution was added to prepare the replicate exposure solutions at nominal concentrations of 7.5, 15, 30, 60 and 120 mg/L. Exposure solutions were stirred with a Teflon coated stir rod.
- Eluate: no
- Differential loading: no
- Controls: water control

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: fathead minnow
- Source: Obtained from Aquatic BioSystems, Inc., Fort Collins, CO, USA (received on 31 May 2000)
- Length at study initiation: 2.76 - 2.79 cm
- Weight at study initiation: 0.32 - 0.34 g
- Feeding during test: none

ACCLIMATION
- Acclimation period: at least 14 d
- Acclimation conditions: same as test
- Feeding frequency: twice daily until 24 h prior to test start

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

120 mg CaCO3/L

Test temperature:

20 - 21 °C

pH:

7.4 - 8.4

Dissolved oxygen:

6.5 - 9.1 mg/L

Nominal and measured concentrations:

Nominal: 0, 7.5, 15, 30, 60, 120 mg/L
Measured: 0, 6.1, 13.6, 28.5, 57.4, 115.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Pyrex glass 30.5-cm cuboidal chromatography jars
- Type: closed with glass lids and sealed with parafilm
- Material, fill volume: glass, fill volume: 20 L, headspace: 2700 cm³
- Renewal rate of test solution: after 48 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Biomass loading rate: 1 g wet weight/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Pumped from a large underground storage reservoir and passed through polypropylene filter tubes, through a series of activated carbon filter tubes and finally through another set of polypropylene filter tubes. The filtered water stream was then treated with sodium thiosulfate via a chemical injection system. The filtered-treated water was then tempered to 20 ± 2 °C by passage through a heat-exchange unit and distributed throughout the testing facility through stainless steel and PVC piping.
- Total organic carbon: 3.2 mg/L
- Metals: < detection limit
- Pesticides: < detection limit
- Chloride: 23.7 mg/L
- Alkalinity: 92.4 mg CaCO3/mL
- Volatile organics: < detection limit
- PCBs: < detection limit
- Conductivity: 290 µmhos/cm
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h dark (30 min transition period)

EFFECT PARAMETERS MEASURED: Mortality was observed every day. Behaviour and appearance were observed throughout the test.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

28.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

40.45 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 28.5 - 57.4 mg/L

Details on results:

- Behavioural abnormalities: No behavioural abnormalities were observed at concentrations of control, 7.5, 15 and 30 mg/L. At the highest test concentration the test organisms showed abnormal behaviour after 4 h. After 24 h the remaining test organisms at 60 mg/L were near death and showed abnormal behaviour.
- Mortality of control: < 10%
- Other adverse effects control: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: The replicates of nominal 7.5, 15 and 30 mg/L were clear and colourless throughout the test. At time 48 h, the nominal 60 mg/L replicate A solution was slightly cloudy. At time 24 h, the nominal 60 mg/L replicate B was very slightly cloudy. At time 48 h, the nominal 60 mg/L replicate B was slightly cloudy. At time 24 h, the nominal 120 mg/L solutions very slightly cloudy.

Reported statistics and error estimates:

Spearman-Karber with EC_LC50.SAS Statistical Program, Ver. 1, Sebaugh's Information Services, Inc, Lake Ozark, USA.

Sublethal observations / clinical signs:

Table 1: Percent mortality and analysed concentrations (mean from the geometric mean of replicates) throughout the study

Nominal exposure [mg/L]	Analyzed exposure [mg/L]	Number of exposed organisms	Percent mortality at each observation
			24 h	48 h	72 h	96 h
Control	not detected	14	0%	0%	7%	7%
7.5	6.1	14	0%	0%	0%	0%
15	13.6	14	0%	0%	0%	7%
30	28.5	14	0%	0%	0%	0%
60	57.4	14	79%	100%	100%	100%
120	115.0	14	100%	100%	100%	100%

Table 2: Results of concentration verification of exposure solutions

Nominal concentration	Analyzed solution concentrations [mg/L]
	Replicate	0 h	48 h (seasoned)	48 h (renewal)	96 h
Control	A	not detected	not detected	not detected	not detected
Control	B	not detected	not detected	not detected	not detected
7.5	A	8.19	4.20	7.64	5.49
7.5	B	7.50	3.57	7.26	6.49
15	A	16.11	10.80	14.37	12.55
15	B	15.99	12.04	15.49	12.63
30	A	31.23	25.68	30.38	27.74
30	B	30.27	25.05	33.03	25.99
60	A	61.36	52.38	not applicable	not applicable
60	B	63.48	53.25	not applicable	not applicable
120	A	114.69	110.61*	not applicable	not applicable
120	B	124.22	111.01*	not applicable	not applicable

* Sampled at time 24 h due to complete mortality.

Description of key information

LC50 (96 h): 40.45 mg/L for Pimephales promelas (OECD 203)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

40.45 mg/L

Additional information

Three experimental studies investigating the short-term toxicity of ethylene diacetate (CAS 111-55-7) to fish are available.

The key study was performed according to OECD 203 (GLP) with the fathead minnow Pimephales promelas (Hirsch, 2000). Test substance concentrations up to 120 mg/L (nominal) were tested in a semi-static exposure regime. The nominal concentrations were measured by GC/FID analysis. Significant effects were observed at the two highest test concentrations, leading to a LC50 (96 h) of 40.45 mg/L (meas. geom. mean. Deviations between nominal and measured concentrations were > 20% throughout the test and therefore, the LC50 is expressed based on measured concentrations.

Two further experimental results are available from literature (Dawson et al., 1977). These studies was not performed according to an internationally accepted guideline but can be compared to OECD 203. Since the documentation of the study design is insufficient, the results were not used for risk assessment and rated reliability 3. No information was provided on how many tests species were used and the number of replicates was not given. Nevertheless, the results are in the same range compared to the key study. The study resulted in a LC50 (96 h) of 90 mg/L for the bluegill sunfish Lepomis macrochirus and a LC50 of 78 mg/L for the tidewater silverside Menidia beryllina.