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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 July - 09 August 2017
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:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Guidance document on aquatic toxicity testing of difficult items and mixtures, OECD series on testing and assessment
Version / remarks:
Document number 23, December 14, 2000.
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
The characterization and stability of the test item have been performed according to cGMP Guidelines

Test material

Constituent 1
Chemical structure
Reference substance name:
1-((2-chlorophenyl)-N-methylcarbonimidoyl)cyclopentan-1-ol
EC Number:
641-894-8
Cas Number:
6740-87-0
Molecular formula:
C13H16ClNO
IUPAC Name:
1-((2-chlorophenyl)-N-methylcarbonimidoyl)cyclopentan-1-ol
Test material form:
solid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Analytical purity: 99.5% (based on assay UPLC)
- Source and lot/batch No.of test material: I17BB0589
- Expiration date of the lot/batch: 12 February 2018 (retest date)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In refrigerator (2-8°C) in container flushed with argon
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: solubility in water: not available

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Sampling method: Samples for possible analysis were taken from all test concentrations and the control at t=0 and t=48h. 2.0 mL of volume was taken. In addition, the filter containing the undissolved residue was kept for possible analysis. At the end of the exposure period, the replicates were pooled at each concentration before sampling.
- Sample storage conditions before analysis: The samples were stored in a freezer. Additionally, reserve samples of 2.0 mL were taken for possible analysis.

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to an expected low water solubility of the test item, saturated solutions (SS) were prepared. The preparation of test solutions started with a loading rate of 100 mg/L applying three days of magnetic stirring at room temperature to ensure maximum dissolution of the test item in test medium. This resulted in a clear and colourless dispersion that contained undissolved floating material and precipitate. The obtained mixture was filtered through a 0.45 µm membrane filter (Whatman; RC55) to remove the undissolved fraction. The filter was pre-conditioned with a small volume of test solution that was discarded. The resulting SS was used as the highest test concentration. Lower test concentrations were prepared by subsequent dilutions of the SS in test medium..
- Evidence of undissolved material (e.g. precipitate, surface film, etc): All final test solutions were clear and colourless

Test organisms

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water-flea
- Source: in-house laboratory culture with a known history
- Age at study initiation (mean and range, SD): < 24 h (from parental daphnids of more than 2 weeks old, maximum age 4 weeks)
- Method of breeding: 250 newborn daphnids (i.e. less than 3 days old) are placed into 5 liters of medium in an all-glass culture vessel. Medium is M7. After 7 days of cultivation half of the medium is renewed twice a week.
- Feeding during cultivation: fresh water algae, daily
- Feeding during test: no

ACCLIMATION
- Acclimation period: not relevant

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h

Test conditions

Hardness:
180 mg/L as CaCO3
Test temperature:
18-22°C
pH:
at t=0h: 8.0-8.7
at t=48h: 8.2-9.0
Dissolved oxygen:
at t=0h: 8.4
at t=48h: 9.0
Salinity:
not relevant
Conductivity:
not specified
Nominal and measured concentrations:
* Range finder test:
Nominal concentrations: 10 and 100% SS,
Measured concentrations (mg/L) at t=0h: 3.66, 35.6
Measured concentrations (mg/L) at t=48h: 2.31, 22.6

* Final test:
Nominal concentrations: 0, 10, 18, 32, 56 and 100% of a SS prepared at 100 mg/L
Measured concentrations (mg/L) at t=0h: n.d., 3.3, 6.22, 10.9, 19.1 and 34.4
Meausred concentrations (mg/L) at t=48h: n.d., 2.18, 4.07, 6.93, 12.6 and 22.3
n.d. = not detected
Details on test conditions:
TEST SYSTEM
- Test vessel: all glass vessel
- Material, size, headspace, fill volume: 60 mL, all glass filled with 50 mL
- Aeration: No aeration of the test solutions
- No. of organisms per vessel: 5 per replicate of each concentration
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Test medium: adjusted ISO medium

OTHER TEST CONDITIONS
- Photoperiod: 16h light daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Immobility was defined as those organisms which are not able to swim within 15 seconds after gentle agitation of the test container and was monitored after 24 and 48 hours of continuous treatment with the test substance.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: x 1.8
- Range finding study
- Test concentrations: 1 and 10% of a SS prepared at a loading rate of 100 mg/L (setup: combined range finder/limit test)
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
yes
Remarks:
Potassium dichromate

Results and discussion

Effect concentrations
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
21 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% C.L.: 16-28 mg/L
Details on results:
- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: none
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Mortality: no
- EC50/LC50: 48-h EC50 = 0.59 mg/L. The sensitivity of the daphnids was within the range determined with the historical data collected at Charles River Den Bosch.
Reported statistics and error estimates:
The 24-EC50 could not be calculated because the observed effects were below 50%.
After 48 hours of exposure, there was no concentration between the highest concentration (A) at
which 0% immobility occurred and the lowest concentration (B) at which 100% immobility occurred.
The 48h-EC50 was calculated as (AB)½, where A and B were set as limits of the 95% confidence interval.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of T003641 to Daphnia magna was determined in a 48 hour static test according to OECD guideline 202. Based on the geometric means of the concentrations, the 48-h EC50 of JNJ54433041-AAA (T00361) was determined to be 21 mg T003641/L with a 95% confidence interval ranging from 16 to 28 mg/L. The results of the test can be considered reliable without restriction.

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