1,1-Bis(acryloyloxymethyl)ethyl isocyanate

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 - 09 Jun 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

Version / remarks:

2011

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Staatliches Gewerbeaufsichtsamt Hildesheim

Test material

Test material information

Constituent 1

Reference substance name:

2-isocyanato-2-methyl-3-(prop-2-enoyloxy)propyl prop-2-enoate

Cas Number:

886577-76-0

Molecular formula:

C11H13NO5

IUPAC Name:

2-isocyanato-2-methyl-3-(prop-2-enoyloxy)propyl prop-2-enoate

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

yes

Remarks:

TOC

Details on sampling:

- Concentrations: All test item concentrations and the control - Sampling method: according to DIN EN 1484 at the start and the end of the exposure

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION - Method: 20 mg test item/L were freshly prepared with dilution water. This stock solution was used to achieve the required test item concentrations.- Evidence of undissolved material: stock solution was agitated until the solution was visually clear

Test organisms

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM - Common name: green algae - Strain: Pseudokirchneriella subcapitata SAG 61.81 - Source: Sammlung von Algenkulturen (SAG) Pflanzenphysiologisches lnstitut der Universitat Gottingen Nikolausberger Weg 18, D-37073 Gottingen, Germany - Age of inoculum: A three days old preculture, was used as inoculum - Method of cultivation: Fresh stocks are prepared every month on Z-Agar. Light intensity amounted to 35-70 μE m-2 s-1 for 24 h per day

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test conditions

Hardness:

0.24 mmol Ca+mg/L

Test temperature:

22.0 - 23.0 °C (mean 22.5 °C)

pH:

7.93 - 8.95 (control)7.73 - 8.95 (test item concentration)

Nominal and measured concentrations:

Control, 0.200, 0.632, 2.00, 6.32 and 20.0 mg/L (nominal)

Details on test conditions:

TEST SYSTEMTest vessel: - Type: open - Material, size, headspace, fill volume: Sterile Erlenmeyer flasks (vol. 250 ml) with cotton wool plugs, 100 mL test volume - Aeration: Test containers were placed on a rotary shaker and oscillated at approximately 70 rpm - Initial cells density: approximately 5 x 10 E+03 - 10^4 cells/mL (corresponds to 6305 cells/mL) - Control end cells density: 1375608 cells/mL - No. of vessels per concentration (replicates): 3 - No. of vessels per control (replicates): 6 GROWTH MEDIUM - Standard medium used: yes TEST MEDIUM / WATER PARAMETERS - Source/preparation of dilution water: according to OECD guideline - Culture medium different from test medium: no - Intervals of water quality measurement: pH was measured at test start and test end, Temperature was measured continously and light intensity was measured prior to test start OTHER TEST CONDITIONS - Sterile test conditions: yes - Photoperiod: 24 hours/day light - Light intensity and quality: approx 4440 to 8880 lux, corresponding to 60 to 120 μE*m-2*s·-1, fluorescent tubes, OSRAM L 36 W/865, cool daylight, within ± 15 % over incubation area - Incubation: The flasks were positioned randomly and repositioned daily EFFECT PARAMETERS MEASURED: cell density via fluorescence - Determination of cell concentrations: Fluorometer: Microplate Reader Chameleon V (HIDEX) with Software Micro Win 4.41 (MIKROTEC LABORSYSTEME GMBH) - Chlorophyll measurement: measured daily via Chlorophyll a fluorescence, excitation at 436 nm, emission at 685 nm TEST CONCENTRATIONS - Spacing factor for test concentrations: dilution factor of √10Range finding study - Test concentrations: Control, 1.00, 10.0 and 100 mg/L - Results used to determine the conditions for the definitive study: The concentration range of the main test was based on the results of the range-finding test

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

6.47 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: 95% Confidence Intervals

Remarks:

6.34 - 6.60 mg/L

Details on results:

- Exponential growth in the control (for algal test): yes - Observation of abnormalities (for algal test): Microscopic evaluation of the cells at the start and end of exposure revealed no morphological abnormalities.

Results with reference substance (positive control):

- ErC50: 0.564 mg/L

Any other information on results incl. tables

Table 1: Biological Results: NOEC, LOEC, EC-values and 95% Confidence Intervals of the test item (0 - 72 hours) based on nominal test item concentrations [mg/L]

	Growth Rate Inhibition Nominal test item concentration [mg/L]
NOEC	0.632
LOEC	2.00
ErC10	2.82 (2.59 - 3.06)
ErC20	3.82 (3.61 - 4.02)
ErC50	6.47 (6.34 - 6.60)
	Yield Inhibition Nominal test item concentration [mg/L]
NOEC	0.632
LOEC	2.00
EyC10	1.65 (1.38 - 1.91)
EyC20	2.01 (1.78 - 2.29)
EyC50	2.99 (2.63 - 3.58)

Table 2: Analytical Results: TOC Content of the Test Media at the Start and the End of the Exposure

Nominal Test Item Concentration [mg/L]	TOC [mg C/L]
	Test start (without algae), 0 hours	Test end (without algae), 72 hours
20.0	11.6	12.3
6.32	2.54	5.45
2.00	< 2.00	2.63
0.632	< 2.00	< 2.00
0.200	< 2.00	< 2.00
Control	< 2.00	< 2.00

Limit of quantification = 2.00 mg C/L

Table 3: EC50-Values of the Reference Item based on nominal concentrations [mg/L], (0-72 hours)

	Current Study	Valid Range (average ± 3 x SD)
	Growth Rate inhibition
ErC50	0.564	0.797 ± 0.428
95 % confidence interval	0.546 - 0.582
	Yield inhibition
EyC50	0.322	0.415 ± 0.275
95 % confidence interval	0.239 - 0.342

SD = Standard deviation

Applicant's summary and conclusion

Validity criteria fulfilled:

yes