[No public or meaningful name is available]

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:

FROM 21 OCT 2022 TO 28 OCT 2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2023

Report date:

2023

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

Adopted April 13, 2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Guidance Document on Aqueous-phase Aquatic Toxicity Testing of Difficult Test Chemicals, OECD series on testing and assessment number 23 (2nd edition)

Version / remarks:

Adopted February 08, 2019

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-[(1-[[6-cyano-5-(trifluoromethyl)pyridin-3-yl]carbamoyl]cyclobutyl)amino]-2-fluoro-N-methylbenzamide

Cas Number:

1950587-20-8

Molecular formula:

C20H17F4N5O2

IUPAC Name:

4-[(1-[[6-cyano-5-(trifluoromethyl)pyridin-3-yl]carbamoyl]cyclobutyl)amino]-2-fluoro-N-methylbenzamide

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: 22CN0611
- Expiration date of the lot/batch: 2023-05-11 (retest date)
- Purity: 99.9%
- Purity test date: 2022-05-31 (certificate of analysis release date)
- Physical appearance: brown powder

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- Stability under storage conditions: not indicated
- Stability under test conditions: not indicated
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: not indicated
- Treatment of test material prior to testing: no treatment

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples for possible analysis were taken from the limit concentration and the control according to the schedule below.

Frequency at t=0 h and t=48 h.
Volume 2.0 mL from the approximate center of the test solutions.
Storage Samples were stored in a freezer (set to maintain -20°C) until analysis at the analytical laboratory of the Test Facility.

At the end of the exposure period, the replicates were pooled at each concentration before sampling.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Preparation of test solutions started with a loading rate of 100 mg/L applying a three-day period of magnetic stirring to ensure maximum dissolution of the test material in medium. Thereafter, the aqueous Saturated Solution (SS) was collected by filtration through a 0.45 µm membrane filter (RC55, Whatman) and used as the limit test concentration.
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): The test solution was clear and colorless at the end of the preparation procedure. Any residual volumes were discarded.

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:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

The hardness of test medium expressed as CaCO3: 180 mg/L

Test temperature:

18-22°C

pH:

At t=0:h 7.8-8.0
At t=48h: 7.8-7.9

Dissolved oxygen:

At t=0h: 8.3 mg/L
At t=48h: 8.7-8.8 mg/L

Salinity:

not relevant

Nominal and measured concentrations:

Combined limit/range finder test
Nominal concentrations: 0 (blank) and 100% of a SS prepared at 100 mg/L
Measured concentrations (mg/L) at t=0h: Measured concentrations (mg/L) at t=48h:
LOQ is 0.07 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 60 mL, glass beaker filled with 50 mL
- Aeration: not during the test
- No. of organisms per vessel: 5
- 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: not applicable (limit test)
- Test concentrations: 100% of a SS prepared at 100 mg/L

RANGE-FINDING STUDY
- Test concentrations: 0, 10 and 100% of a SS prepared at 100 mg/L
- Results used to determine the conditions for the definitive study: no immobility was observed in the control and the test concentrations throughout the exposure period

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.48 mg/L

Nominal / measured:

meas. (geom. mean)

Remarks on result:

not determinable

Details on results:

- Immobility of control: none
- Other adverse effects control: no
- Abnormal responses: none

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: The batch of Daphnia magna tested, showed expected sensitivity to Potassium dichromate

Reported statistics and error estimates:

the 24 and 48-h EC50 could not be calculated because no adverse effects were observed in this study

Any other information on results incl. tables

Samples taken from the limit concentration and the control were analyzed. The measured concentration at the start of the test was 0.52 mg/L, for the limit test concentration. During the exposure period, the concentrations remained stable, i.e., were at 86% relative to initial at the end of the test.

Based on these results, the geometric mean concentration of the limit test concentration was calculated to be 0.48 mg/L and used to express effect parameters.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, the 48h-EC50 for Daphnia magna exposed to JNJ-63632335-AAA (T003671) was beyond the range of concentrations tested, i.e., exceeded a geometric mean concentration of 0.48 mg/L.