3-Isocyanatomethyl-3,5,5-trimethylcyclohexyl isocyanate, oligomers, 2-Hydroxy-1(2)- methylethyl carbamate blocked

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study without restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-(carbamoyloxy)propyl N-[(3-{[3,5-bis({5-[({[2-(carbamoyloxy)propoxy]carbonyl}amino)methyl]-1,3,3-trimethylcyclohexyl}methyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]methyl}-3,5,5-trimethylcyclohexyl)methyl]carbamate

EC Number:

921-910-2

IUPAC Name:

2-(carbamoyloxy)propyl N-[(3-{[3,5-bis({5-[({[2-(carbamoyloxy)propoxy]carbonyl}amino)methyl]-1,3,3-trimethylcyclohexyl}methyl)-2,4,6-trioxo-1,3,5-triazinan-1-yl]methyl}-3,5,5-trimethylcyclohexyl)methyl]carbamate

Test material form:

solid

Details on test material:

- Analytical purity: 83.9 g/100g
- Composition of test material, percentage of components: 0.2g/100g water, 13.4g/100g n-butyl acetate, 2.5g/100g 1,2-propanediol, monocarbamate
- Lot/batch No.: 389-48
- Expiration date of the lot/batch: 2019-05-23

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The definitive test was performed using the single filtrate of the test item loading rate of 120 mg/L plus
the control
The appropriate amount of the test item was weighed in a glass crystalliser, quantitatively transferred
into a volumetric flask and filled up with the corresponding volume of the AAP medium (Table 3)
[SOP/W/7].
The mixture was visually heterogeneous with undissolved particles. Therefore, the mixture and the
control (the AAP medium, 1500 mL) were mechanically shaken for 48 h (room temperature, 90
revolutions per minute) and then stored at room temperature for a settling phase (no shaking). After
24 h of settling phase, the mixture was visually heterogeneous, with undissolved particles. Therefore,
the mixture and the control were filtered through a conditioned nitrocellulose membrane filter (0.45 Sm,
Millipore), [SOP/W/37]. This is a WAF-approach followed by filtering, which results in a water soluble
fraction [9]. Each filtrate was visually homogeneous and transparent.
At exposure initiation, four samples (without algae) of the filtrate of the test item loading rate of
120 mg/L and the filtrate of the control were collected. One of them was transferred for chemical
analyses at exposure initiation, whereas the others were stored under test conditions and transferred
for chemical analyses one after the other every 24 h of exposure (24, 48 and 72 h old). [SOP/W/83].
200 ml of each filtrate was used for the saturation of the inner walls of the test vessels and then
discarded.
An additional replicate without the algae was used as a background for spectrophotometric
measurements.
The cell density in the four-day-old algal pre-culture was determined by counting the number of cells in
the Bürker chamber under a microscope. It was 2.9 x 106 cells/mL. Each filtrate was inoculated with
algal pre-culture to obtain the initial algal density of 0.01 x 106 cells/mL. Each treatment was split up
into six replicates.

Test organisms

Test organisms (species):

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

Details on test organisms:

TEST ORGANISM
- Common name: green algae
- Strain: SAG 61.81
- Source (laboratory, culture collection): Algae Collection University of Göttingen, Germany, cultivated at the performing institute.
- Method of cultivation: Algae were transferred from agar bevels to the fresh medium (for Pseudokirchneriella subcapitata AAP medium is recommended, Appendix 2) in 250 mL Erlenmeyer flask and incubated in 21 – 24 C in constant illumination (pre-culture) [1], [SOP/W/65]. The algae culture was renewed (inoculated to a new medium) twice a week in sterile conditions [SOP/W/56, SOP/W/70].
A pre-culture with cell density of 1 x 104 per mL was renewed three days before the definitive test. The pre-culture was used for inoculation of the test concentrations and the control.

ACCLIMATION
- Culturing media and conditions (same as test or not): yes
- Any deformed or abnormal cells observed: no

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Test conditions

Hardness:

20.3 mg/L as
CaCO3

Test temperature:

21.7 – 22.2°C

pH:

pH of the control: 7.45 – 8.95

Conductivity:

94.5 μS/cm

Details on test conditions:

TEST SYSTEM
- Test vessel: glass Erlenmeyer flasks 250 mL
- Type (delete if not applicable): closed with air permeable stoppers
- Material, size, headspace, fill volume: 100 mL
- Aeration: none
- Initial cells density: 10E+04 cells/mL
- Control end cells density:
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 2

GROWTH MEDIUM
- Standard medium used: yes

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16:8 light:dark
- Light intensity and quality:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: spectrophotometric measurement

TEST CONCENTRATIONS
- Range finding study: yes

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

EL50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

EL10

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Applicant's summary and conclusion

Validity criteria fulfilled:

yes