2-Propenoic acid, 2-isocyanatoethyl ester

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The experimental start and completion dates of the study were 4 June 2007 and 16 April 2008, respectively.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

from MHRA (UK GLP Monitoring authority)

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

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: At each concentration, the test substance (85.4, 187.8, 414, 910 or 2000 mg) was dissolved in diluent water (5 l). The contents of each flask were shaken vigorously before being poured into a test vessel (glass aquarium) containing diluent water (15 litres).
- other: Following a review of the data , the results of the prelimenary tests where the fish had been placed in the test vessels either before or immediately after the test substance was added were considered unrepresentative because of the highly unstable nature of AOI following its addition to water. Consequently, it was decided to follow the guidance given in OECD Monograph Number 23 for the testing of unstable compounds. As AOI had a half-life of less than one hour, the test substance was allowed to degrade after its addition to water before the fish were added.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: The fish were reared at Huntingdon Life Sciences (Eye Research Centre) from eggs and milt supplied by a commercial fish farm in the UK on 21 December 2007
- Age at study initiation:about 3 month
- Length at study initiation: 5.81 cm
- Weight at study initiation: 2.04 g
- Method of breeding: The fertilised eggs and hatched fry were held in an aerated supply of diluent water under flow-through conditions until use.


ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions: The fertilised eggs and hatched fry were held in an aerated supply of diluent water under flow-through conditions until use. During the 14-day period immediately before the definitive test, temperatures remained within the range 11.8 to 14.7 °C, pH values within the range 7.69 to 7.95, dissolved oxygen concentrations within the range 92 to 102 % air saturation value (ASV) and total hardness within the range 164 to 170 mg/l as CaCO3.
- Type and amount of food: Commercial fish food (TROUW (UK) Ltd; Nutra Fry 02) equivalent to between 1 and 1.5 % of the total wet-weight of fish in the holding tank during the 14 days before the start of the definitive test.
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): No medication was given during the holding period, and mortalities were recorded as 3.5% in the 14 days before the definitive test.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

154 - 162 mg/L as CaCO3

Test temperature:

12.1 - 15.2 °C

pH:

7.11 - 8.16

Dissolved oxygen:

91 - 106 % os saturation

Nominal and measured concentrations:

Nominal concentrations: 4.27, 9.39, 20.7, 45.5 and 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, fill volume: glass aquarium, contained 20 litres of medium to a depth of 18 cm.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.71 g bodyweight/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to guideline (dechlorinated tap water, hardness 154 mg/l as CaCO3)


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light: 8 hours dark

EFFECT PARAMETERS MEASURED: In addition to observations on mortality at approximately 2, 4, 24, 48, 72 and 96 hours,
subjective assessments were also made on the incidence and type of any sub-lethal effects compared with control fish.

TEST CONCENTRATIONS
- Range finding study: The study comprised of five range finding tests, two trials and a definitive test with five test concentrations and a dilution medium control group.

- Test concentrations: 0.1, 1.0, 10, 50 and 100 mg/L in differend range finding set ups. AOI was very unstable in water and that it was not possible to achieve the nominal concentration of the parent material. The second trial was conducted to determine if it was possible to prepare a solvent stock solution of AOI at 50 mg/l for use in a toxicity test. The data showed that this was feasible.

- Results used to determine the conditions for the definitive study:
In the first range finding test, AOI was dissolved in dimethyl formamide (DMF) before its addition to water and nominal concentrations of 0.1, 1 and 10 mg/l were employed; no mortality or adverse effects on the fish were observed after 96 hours. The second range finding test was conducted without the use of solvent so that fish could be exposed at nominally 100 mg/L. The fish were placed in the test vessel before the test substance was added and they died within 15 minutes of adding the test substance. Following the death of the fish at 100 mg/l, the third range finding test employed nominal concentrations of 1, 10 and 50 mg/l. At each concentration, the test
substance was dissolved in DMF and immediately following its addition to water, the fish were added. In the fourth and fifth range finding tests, fish were exposed to AOI at nominal concentrations of 100 and 10 mg/l, respectively, following the direct addition of AOI to
diluent water. The fish were placed in the test media between 40 and 60 minutes after dosing and all fish died within two hours at 100 mg/l but no mortality occurred at 10 mg/l after 72 hours.
Based on the results of the first, fourth and fifth range finding tests, the definitive test was conducted at nominal concentrations of 4.27, 9.39, 20.7, 45.5 and 100 mg/l.Because of the highly unstable nature of AOI following its addition to water. Consequently, it was decided to follow the guidance given in OECD Monograph Number 23 for the testing of unstable compounds (see also test substance preparation).

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

30.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

20.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Any other information on results incl. tables

Sublethal observations / clinical signs:

The 96-hour LC50 value for AOI with rainbow trout was 30.7 mg/l, with 95% confidence

limits of 20.7 and 45.5 mg/l. The “no observed effect concentration” was 20.7 mg/l.