2-[[(butylamino)carbonyl]oxy]ethyl acrylate

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:

18 May - 18 Sep 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

adopted 17th July 1992

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

No 440/2008 of 30 May 2008

GLP compliance:

yes (incl. QA statement)

Remarks:

Swiss Federal Office of Public Health

Analytical monitoring:

yes

Details on sampling:

The test concentrations were determined by HPLC prior to the addition of the fish to the test solutions and after 2, 24, 48, 72 and 96 h of exposure.

Vehicle:

no

Details on test solutions:

Since the test item is water soluble, the test concentrations were prepared by dilutions of a stock solution in natural water.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: Commercial supplier (Aquarium Stern, 4310 Rheinfelden).
- Length at study: 1.6 - 2.0 cm
- Weight at study: 26 - 164 mg

ACCLIMATION
- Acclimation period: Fish were gradually acclimatized to the test conditions and were held for at least an additional 12 days prior to testing
- Acclimation conditions: Water of similar temperature (23±2.0 °C) and quality used in the test
- Health/mortality: No mortality was observed

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Total Hardness: 206 - 242 mg/L
CO3 Hardness: 186 - 217 mg/L
Residual Hardness: 20 - 30 mg/L

Test temperature:

22 °C

pH:

6.6 - 8.6

Dissolved oxygen:

6.7 - 7.2 mg/L

Nominal and measured concentrations:

Test concentrations nominal: 0, 0.625, 1.25, 2.5, 5.0 and 10 mg/L
Test concentraions measured (geometric mean): 0, 0.182, 0.573, 1.6, 4.1 and 8.45 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria of 2 l (14.5 x 14.0 x 9.5 cm) filled with 1.0 liter of water.
- Aeration: Yes
- No. of organisms per vessel: 7 fish
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- Biomass loading rate: 0.385 -0.717 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Natural water of the town of Schlieren
- Nitrate: 7.3 mg/L
- Chloride: 16.1 mg/L
- Sulfate: 18.6 mg/L
- Conductivity: 441 - 631 µS/cm
- Culture medium different from test medium: No

OTHER TEST CONDITIONS
- Photoperiod: 16 h photoperiod a day
- Light: White fluorescent tubes

EFFECT PARAMETERS MEASURED (with observation intervals) :
- Oxygen, pH, temperature: Prior to the addition of the fish to the test solutions and after 2, 24, 48, 72 and 96 h of exposure.
- Observations of the fish: Mortality of the fish was recorded after 2, 24, 48, 72 and 96 h of exposure.

RANGE FINDER
- Test concentrations: 6.0 mg/L (limit test)
- Results used to determine the conditions for the definitive study: In this limit-test, three fish died after 72 h and three further fish died after 96 h of exposure, yielding a mortality rate of 86% after 96 h. Consequently, a full test with 5 concentrations was performed.

Reference substance (positive control):

not specified

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.52 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% confidence limits: 1.91–3.31 mg/L

Details on results:

- At the nominal concentrations of 10.0 and 5.00 mg/L all 7 fish died within 24 and 48 h of exposure, respectively.
- LC100(96h): 5.00 mg/L nominal concentration, which corresponds to 4.10 mg/L measured concentration
- No mortality was observed at all other nominal concentrations and in the blank control after 96 h of exposure.
- NOEC (96h): 2.50 mg/L nominal, which corresponds to 1.60 mg/L measured concentration
- Behavioural abnormalities: None

Sublethal observations / clinical signs:

The analytical results indicated that the concentrations were decreasing over time. Therefore, the statistical estimation of the effective concentrations was based on the geometric mean of the measured concentrations over the whole relevant test period.

Table 1: Analytical determination of the test concentrations by HPLC (mg/L):

Nominal concentration (mg/L)	0h	2h	24h	48h	72h	96h	GM 0-96h
0.625	0.593	0.562	0.475	0.342	0.068	0.010	0.182
1.25	1.54	1.50	1.45	1.11	0.05	0.20	0.573
2.50	2.05	2.12	1.89	1.47	1.12	1.12	1.60
5.00	4.5	4.63	3.93	3.46	*	*	4.10
10.0	8.67	8.80	7.90	*	*	*	8.45

* All fish died, no further observations

Table 2: Cumulative number of dead fish

Nominal concentration (mg/L)	24h	48h	72h	96h
0 (blank)	0	0	0	0
0.625	0	0	0	0
1.25	0	0	0	0
2.50	0	0	0	0
5.00	0	7	*	*
10.0	7	*	*	*

*All fish died, no further observations

Description of key information

LC50 (96 h) = 2.52 mg/L (measured geometric mean, OECD 203, D. rerio)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

2.52 mg/L

Additional information

There is one key GLP study available investigating the acute lethal toxicity of the substance to fish according to the OECD guideline 203.

In a static test D. rerio were exposed to the nominal test item concentrations 0.625, 1.25, 2.50, 5.00 and 10.0 mg/L, which were derived from a previous limit test where 86% mortality had been observed at 6.00 mg/L. The test concentrations were measured by HPLC at the beginning of the test and after 2, 24, 48, 72 and 96 h of exposure. Since the test substance is water soluble, the test concentrations were prepared by serial dilutions of a stock solution in natural water.

The analytical results indicated that the concentrations of the test substance decreased over time. Therefore, the statistical estimation of the effect concentrations was based on the geometric mean of the measured concentrations over the entire tested time period.

All 7 fish died within 24 and 48 h at the nominal concentrations of 10.0 and 5.0 mg/L, respectively. No mortality was observed at all other nominal concentrations and in the blank control after 96 h of exposure. Based on these data, the calculated LC50 (96 h) was 2.52 mg/L with respect to the measured concentrations. The NOEC (96 h) was 2.50 mg/L (nominal), which corresponds to a measured concentration of 1.60 mg/L. The LC100 (96 h) was 5.00 mg/L (nominal), which corresponds to a measured concentration of 4.10 mg/L.