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

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

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

Specific details on test material used for the study:

- Name of test material (as cited in study report):
ZQ542226, Batch: 389-48, Sample No: 3, Date of manufacture:
23 May 2016, Expiry date: 23 May 2019.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Analyses were performed at exposure initiation in samples of freshly prepared solutions, at the first
renewal in samples of 24 hour old solutions and at exposure termination in samples of 96 hour old
solutions.

Test solutions

Vehicle:

no

Details on test solutions:

The definitive test was performed with the filtrate of a loading of 120 mg/L and the filtrate of the
control. The test was performed under a semi-static test design (i.e. with daily renewals) and with a
WAF (Water Accommodated Fraction) approach.
The weighed amount of the test item was added to the test medium (analytical balance, Ohaus,
PA214CM/1) [SOP/W/7]. The resulting mixture of a loading of 120 mg/L was heterogeneous with
visible non-dissolved particles. Therefore, the mixture of a loading of 120 mg/L and, separately, the
control, were mixed for the next 48 hours at room temperature using a mechanic shaker (at 90
rounds per minute). Next, the mixture of a loading of 120 mg/L and, separately, the control, were
incubated at room temperature for 24 hours without shaking. Next, each of the mixtures of a loading
of 120 mg/L and the control was separated by filtration through a conditioned nitrocellulose
membrane filter (Millipore 0.45 Vm HAWG, Merck) under vacuum [SOP/W/37]. The collected filtrates
were visually homogeneous, transparent and used for the exposure as water soluble fraction WSF
[8].
For the filtration, the conditioning was done by pre-filtering a small volume of each test item loading
or the control to saturate the filter (and disposing it) before filtering the volumes for exposure.
Ten fish were introduced into every glass test vessel with 4 L of the filtrate of a loading of 120 mg/L
and the filtrate of the control, respectively. One replicate of every filtrate was used for exposure (i.e. a
total of 2 test vessels). Daily renewals of every filtrate were done during the 96 hour exposure. On
every renewal, all fish were transferred from the old into the respective freshly prepared filtrate.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The test organism was the zebrafish (Danio rerio Hamilton-Buchanan). Adult fish originated from the
laboratory zebrafish breeder named “the International Institute of Molecular and Cell Biology in
Warsaw”. They were admitted, quarantined and kept in the Laboratory of Aquatic Toxicology, Institute
of Industrial Organic Chemistry, Branch Pszczyna.
In the definitive test, the fish were approximately 15 months old, healthy and in good physiological
condition. The health of fish and the hygiene in the animal facility is monitored daily on a regular
basis, according to the animal monitoring and veterinary care programme [SPO/W/88, SOP/W/89].
Before exposure initiation in the definitive test, the fish adaptation to the test conditions lasted for 12
days [1], [SOP/W/23].

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

239 mg/L as CaCO3

Test temperature:

22.3 - 23.2 °C

pH:

7.56 – 7.75

Dissolved oxygen:

92 - 98% of air saturation value

Conductivity:

553 µS/cm

Nominal and measured concentrations:

- Nominal loading rates: 0 (control), 120 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass vessels
- Type: Closed; vessels were covered with glass lids in order to minimise evaporation and to prevent accidental loss of fish (due to jumping out) and accidental contamination. The bottom of glass vessels with fish was coloured but the walls were transparent to facilitate observations during exposure.
- Fill volume: 4 L
- Aeration: Continuous; the gentle bubbling aeration was supplied with a glass tube to the bottom and served also as the orientation reference for the fish.
- Renewal rate of test solution: Daily; on every renewal all alive fish were transferred from the spent solution to the respective freshly prepared solution using a fish-net catcher.
- No. of organisms per vessel: 10
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- Fish loading: 0.28 g/L
- Feeding during test: no


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituent water (ISO 6341 : 1982) was prepared based on deionized water by adding appropriate amounts of stock solutions of reagent grade chemicals. See 'Details test organism' for the preparation method. The composition is tabulated in 'Any other information on materials and methods incl. tables'.
- Culture medium different from test medium: No
- Intervals of water quality measurement: Twice daily (before and after renewal)

OTHER TEST CONDITIONS
- Photoperiod: 16 h light: 8 h darkness
- Light type: fluorescent

EFFECT PARAMETERS MEASURED
During the test observations for mortality and for symptoms of intoxication were conducted and recorded after 3, 6, 24, 48, 72 and 96 h of exposure. These were observations for loss of equilibrium, changes in swimming behaviour, respiration or pigmentation. The fish are considered dead if there is no reaction to external stimuli (touching of the caudal peduncle).

OTHER MEASUREMENTS
In both tests temperature was continuously recorded using an electronic device with a sensor submerged in the glass vessel containing the control (HI 141 thermologger, Hanna Instruments). The pH values and dissolved oxygen concentrations were measured in every freshly prepared test item loading rate (i.e. filtrate) and the control at exposure initiation and on every renewal as well as in every spent test item loading rate and the control on every renewal and at exposure termination in each glass vessel with fish (inoLAB Level 3 pH-Oxi-meter, WTW, Germany). In the test at exposure initiation conductivity was measured in the control (Multi 3430, WTW, Germany and the total hardness was determined by complexometric titration with EDTANa2 (ethylenediamine-tetraacetic acid disodium salt) (digital titration apparatus Solarus, Hirschmann Laborgeräte GmbH&Co., Germany.

PRELIMINARY NON-GLP TEST
- Test loading rates: 0 (control), 5 (diluted), 10 and 100 mg/L.
- No. of organisms per vessel: 5
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- Results used to determine the conditions for the definitive study: 40% mortality was observed after 96 exposure in 100 mg/L loading rate treatment. The test item loading rate used in the definitive test was determined on the basis of the preliminary test results.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes