2-Propenoic acid, 2-isocyanatoethyl ester

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

23 Feb 2011 – 21 Mar 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

adopted 2008

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

482-140-6

EC Name:

-

Cas Number:

13641-96-8

Molecular formula:

Hill formula: C6 H7 N O3 CAS formula: C6 H7 N O3

IUPAC Name:

2-isocyanatoethyl prop-2-enoate

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sampling method: All test solutions were taken from all test solutions at the beginning (0 d), immediately prior to the first renewal of the test solution (1 d) and weekly thereafter (10, 21 d).The highest and lowest test conditions were analyzed after pre-treatments.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test solution of AOI (100 mg/L) was prepared by directly adding 0.010 g of the test item into 100 mL test water. Ultrasonication for 30 min was used to facilitate the dispersion of the test substance. Other test solutions with lower concentrations were prepared by diluting the solution of 100 mg/L.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Age at study initiation (mean and range, SD): The test species were 6 to 24 hours old at the start of the test.
- Source: All organisms used for a particular test originated from cultures established from the same healthy stock of daphnia.
- Amount: 0.2 – 0.4 mg C/Daphnia
- Frequency: Once every two days.

ACCLIMATION
- Acclimation conditions (same as test or not): The stock animals were maintained in culture conditions (light, temperature, and medium) similar to those to be used in the test.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Test conditions

Hardness:

see Table 1 in "Any other information on results incl. tables"

Test temperature:

20 ± 1 °C

pH:

7.4 – 7.5

Dissolved oxygen:

85 – 98%

Nominal and measured concentrations:

Nominal: 0.1, 0.3, 0.9, 27, 8.1 mg/L
Measured: 0.0773 (nominal 0.1 mg/L), 5.47 (nominal 8.1 mg/L) mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass beakers
- Fill volume: 50 mL
- Aeration: None
- Renewal rate of test solution (frequency/flow rate): The test medium was renwed every 24 hours.
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10
- Biomass loading rate: 50 mL/daphnid

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water which had been dechlorinated for at least 24 h was used.
- Culture medium different from test medium: No
- Intervals of water quality measurement: Besides, oxygen concentration, temperatue, hardness and pH values were measured once a week, in the control(s) and in the highest test substance concentration.

OTHER TEST CONDITIONS
- Photoperiod: 16-hour light and 8-hour dark cycle daily
- Light intensity: Light intensity: 1000 to 1500 lux
- Temperature: 20 ± 1 °C
- Oxygen concentration: 85 – 98%

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Observations: The number of living offspring was counted for each vessel daily. The offspring produced by each parent animal were preferably removed and counted daily from the appearance of the first brood to prevent them consuming food intended for the adult. Time to production of first brood; number of broods per animal.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: The following nominal concentrations of AOI were tested in range-finding test: 0.1 ,1 ,10 ,100 mg/L.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

> 8.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 2

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

2.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 3

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Mortality of control: During the test, all animals in the control were alive and the mean number of live offspring produced per parent animal surviving at the end of the test was >60. The coefficient of variation around the mean number of living offspring produced per parent animal in the control(s) was ≤ 25%.
- Other: Compared with the reproduction of the controls, no significant differences were found in 0.1, 0.3 and 0.9 mg/L treatments (p<0.05). However, reproductions of 2.7 and 8.1 mg/L treatments were inhibited significantly by 10.8% and 21.84% respectively (p<0.05).

Results with reference substance (positive control):

No reference substance was used

Reported statistics and error estimates:

Effect concentrations were calculated using One-Way ANOVA

Any other information on results incl. tables

Table 1: Stability test results of AOI in test media

 

Nominal Concentration (mg/L)	Actual concentration (mg/L(TOC))	Analyzed concentration (mg/L(TOC))
		Day 0	Day 1		Day 10		Day 21
		fresh	aged	fresh	aged	fresh	aged
0	-	0.692	0.747	0.811	0.699	0.733	0.595
0.1	0.0773	0.797	0.790	0.800	0.805	0.797	0.796
8.1	5.47	6.22	6.33	5.89	6.14	6.22	6.31

 

 

Table 2: Daphnia adult mortality

 

Nominal concentration (mg/L)	Exposure time (d)
	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
	No. of dead adults
0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
0.1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
0.3	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
0.9	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2.7	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
8.1	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0	2	3	1	0	0

 

 

Table 3: Reproduction inhibition effects

 

Nominal con. (mg/L)	Initial No. of adults	21 d
		Mean fecundity (No. of live offspring per adult)	Inhibition rate
0	10	61	-
0.1	10	59	3.26%
0.3	10	61	1.14%
0.9	10	57	6.51%
2.7	10	55	10.8%
8.1	10	56	8.31%
LOEC (by One-Way ANOVA)		2.7 mg/L
NOEC (by One-Way ANOVA)		0.9 mg/L
EC50		> 8.1 mg/L (nominal concentration)

 

 

Table 4: Validity criteria for OECD 211

 

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality of the parent animals in the control (femaleDaphnia) does not exceed 20% at the end of the test.	During the test, all animals in the control were alive.	yes
The mean number of living offspring produced per parent animalsurvivingat the end of the test is ≥ 60 in the control.	The mean number of live offspring produced per parent animal surviving at the end of the test was 61.	yes

Note: The same validity criterion (20%) can be used for accidental and inadvertent parental mortality for the controls as well as for each of the test concentrations.

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

See Table 4 in "Any other information on results incl. tables"

Conclusions:

Under semi-static conditions of 24 h-renewal, the effect on the reproductive output of Daphnia magna exposed to the test substance for 21 days was assessed according to OCED 211. Under valid semi-static test conditions (24h-renewal), the LOEC is 2.7 mg/L (nominal concentration). And thus the NOEC is 0.9 mg/L (nominal concentration). The EC50 is greater than 8.1 mg/L (nominal concentration).