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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-06-08 until 2017-08-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures
Version / remarks:
Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000
Deviations:
no
Qualifier:
according to guideline
Guideline:
other:
Version / remarks:
Series on Testing and Assessment No. 126, “Short Guidance in the Threshold Approach for Acute Fish Toxicity”, May 31, 2010
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: SANCO/3029/99 rev.4 11/07/00
Deviations:
no
Principles of method if other than guideline:
Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
2-({[3-(isocyanatomethyl)-3,5,5-trimethylcyclohexyl]carbamoyl}oxy)ethyl prop-2-enoate
EC Number:
815-462-6
Cas Number:
124451-79-2
Molecular formula:
C17H26N2O5
IUPAC Name:
2-({[3-(isocyanatomethyl)-3,5,5-trimethylcyclohexyl]carbamoyl}oxy)ethyl prop-2-enoate
Test material form:
liquid
Details on test material:
2-({[3-(isocyanatomethyl)-3,5,5-trimethylcyclohexyl]carbamoyl}oxy)ethyl prop-2-enoate from Evonik Resource Efficiency GmbH, Batch SB 4663

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
The samples were taken from the biological phase of the study. Duplicate samples from the freshly prepared test media of the only test concentration and the control were taken at the start of the test and at day 3. For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test medium renewals, duplicate samples from the aged test media of the only test concentration and the control were collected at day 1 and at the end of the test (after 96 hours) from the approximate centre of the aquaria

Test solutions

Vehicle:
no
Details on test solutions:
Due to the low water solubility of the test item Water accommodated Fractions (WAF) with nominal loading rate of 100 mg test item/L (corresponding to a
mean measured concentration of 42.3 mg test item/L)
and a control was tested.
The control has been made of reconstituted water (see composition below)
Reconstituted Water (ISO Medium):
In deionised water (conductivity <= 10 µScm-1) analytical grade salts were added to following nominal concentrations:
CaCl2 x 2H2O : 2.0 mmol/L (= 294.0 mg/L)
MgSO4 x 7H2O : 0.5 mmol/L (= 123.0 mg/L)
NaHCO3 : 0.75 mmol/L (= 65.0 mg/L)
KCl : 0.075 mmol/L (= 5.8 mg/L)
Water Hardness : 2.5 mmol/L (= 250.0 mg/L) as CaCO3
Alkalinity : 0.8 mmol/L
Ratio of Ca : Mg = 4 : 1 (based on molarity)
Na : K= 10 : 1 (based on molarity)

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Juveniles of Rainbow trout (Oncorhynchus mykiss)
The mean body length of the fish* in the test was 5.03 cm ± 0.2 cm (Mean ± SD), the mean body wet weight 1.07 g ± 0.14 g (Mean ± SD).
* 10 male and female fish from the test fish batch were measured before the start of the test.
The test fish were obtained from Forellenzuchtbetrieb Störk, 88348 Bad Saulgau, Germany.
The fish were obtained and held in the laboratory for at least 12 days before the start of the test. They were held in water of the same quality as used in the test for at least seven days immediately before testing.

Study design

Test type:
semi-static
Water media type:
other: reconstituted water nach ISO as described above
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
2.5 mmol/L (= 250.0 mg/L) as CaCO3
Test temperature:
13.8 to 16.0 °C

Temperature [°C] in the Test Media (presented are mean measured temperature during the study period)
Water temperature [°C]
Water temperature [°C]

[mg test item/L] 0 h 24 h 48 h 72 h 96 h
Control new 15.0 15.8 16.0 15.8 -
aged - 15.8 16.0 15.8 13.8
42.3 new 15.3 15.3 16.0 16.0 -
aged - 15.8 16.0 16.0 13.9
pH:
7.3 to 8.0

Dissolved oxygen:
The dissolved oxygen concentration in the test media did not fall below 88 % of air saturation value during the test
Conductivity:
In deionised water (conductivity <= 10 µScm-1)
Nominal and measured concentrations:
Nominal conentration: 42.3 mg test item/L
The analytical method is not specific to the analyte. Carbon in the test medium and controls was present in insignificant amounts and was not subtracted from
the samples
Details on test conditions:
Type and Size of test vessel: 12 litre glass aquaria with 10 litre test medium
Identification: Each test unit was uniquely identified with serial number and treatment.
Test Environment: Controlled environmental room
Measurement of pH-Values, Dissolved Oxygen and Water Temperature: The water temperature, pH-values and the dissolved oxygen concentrations were determined daily in the freshly prepared and aged test media of each treatment group.
Water Temperature: 13.8 to 16.0 °C
pH-Values of the Test Water: 7.3 to 8.0
Dissolved Oxygen Concentrations: 88 to 100 % of the air saturation value
Light Regime: 16 h light : 8 h dark; 30 min dawn/dusk period was provided
Light Intensity: 270 to 360 lux
Aeration of the Test Water: The test media were slightly aerated during the test.
Recording: Test conditions were recorded with suitable instruments and documented in the raw data.

Control: In the control, test water was used without addition of the test item.
Dosage of Test Item: Before test start and at each test medium renewal, supersaturated stock suspensions of the test item were prepared by suspending test item in test water at concentrations of 100 mg test item/L. The suspensions were carefully stirred in the dark for approximately 48 hours to dissolve as much test item as possible. After stirring followed a settling period for one hour to allow phase separation. Then, the test solutions were extracted from below the water surface. The test media were prepared just before introduction of the fish (= start of the test) and before each test medium renewal.

Introduction of Fish: At the start of the test 7 fish were introduced into each aquarium in a random order.
Number of Replicates: The test was performed with one replicate per treatment group.
Fish per Replicate: 7
Loading: < 1 g fish/L test water
Exposure Time: 96 hours
Test Procedure: A semi static test with daily test medium renewal was chosen to keep the concentrations of the test item in the test media as constant as possible during the test period of 96 hours. During this semi static test the surviving fish were placed in a clean aquarium with freshly prepared test medium of the corresponding concentration every day. The test media were slightly aerated.
Feeding: None

Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Key result
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 42.3 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
>= 42.3 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
The toxic effect of the test item to Rainbow Trout (Oncorhynchus mykiss) was assessed in a semi-static limit test. Based on the test results the 96-hour LC50 was determined to be
> 42.3 mg test item/L based on mean measured concentrations. The NOEC was determined to be ≥ 42.3 mg test item/L also based on mean measured concentration.
Results with reference substance (positive control):
No positive control has been used

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
In this study, the test substance was found to have no effect on the vitality of Rainbow Trout (Oncorhynchus mykiss) after 96 hours at a nominal test item concentration of 42.3 mg/L.
In conclusion, the 96-hour LC50 was determined to be > 42.3 mg test item/L based on mean measured concentrations. The NOEC was determined to be ≥ 42.3 mg test item/L also based on mean measured concentration.



Executive summary:

The acute toxicity of the test substance to fish (Rainbow Trout:Oncorhynchus mykiss)was assessed in a semi-static limittestaccording to OECD Guideline for Testing of Chemicals, Section 2, No. 203, "Fish, Acute Toxicity Test", adopted July 17, 1992 and OECD Series on Testing and Assessment, No. 23, "Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures", December 15, 2000 from 2017-06-23 to 2017-06-29 at the test facility. Measured endpoints were mortality and sublethal effects. A threshold approach was applied.

An acute toxicity test under semi-static conditions with the nominal threshold concentration of 42.3 mg/L was carried out. The selection of the test concentration was based on the derivation of a threshold concentration from the results of a daphnia toxicity test (48-h ErC50 = 45.8 mg test item/L; WAF with a loading rate of nominal 100 mg/L (provided by the sponsor) and an algae acute toxicity test (72-h ErC50: > 45.5 mg test item/L; WAF with a loading rate of nominal 100 mg/L. The duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control. The water quality parameters (temperature, pH-value and oxygen saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits.

If no mortality occurred in the limit test using the threshold concentration, the threshold concentration might be used as a surrogate LC50 value in the further hazard or risk assessment.

The quantification of the test item and / or its degradation products in the test samples was performed by analyzing total organic carbon (TOC).

In the freshly prepared test media at test start and at renewal of the test media, 42 % of the nominal test concentration was found (average of duplicate samples). 42 % of the nominal concentration was also determined in the aged test media after 24 and 96 hours (average of duplicate samples). All reported results refer to mean measured concentrations.