1-amino-N-[6-cyano-5-(trifluoromethyl)-3-pyridyl]cyclobutanecarboxamide

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 28 OCT 2022 TO 09 NOV 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:

1-amino-N-[6-cyano-5-(trifluoromethyl)-3- pyridyl]cyclobutanecarboxamide

Cas Number:

1950587-17-3

Molecular formula:

C12H11F3N4O

IUPAC Name:

1-amino-N-[6-cyano-5-(trifluoromethyl)-3- pyridyl]cyclobutanecarboxamide

Test material form:

solid: particulate/powder

Details on test material:

- Name of test material: JNJ-63632283-AAA (T003665)
- Physical state: solid (powder)
- Appearance: white powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: 22BN0551
- Purity: 98.3%
- Physical appearance: off-white powder
- Expiration date of the lot/batch: 2023-04-28 (retest date)
- Purity test date: 2022-05-13 (certificate of analysis release date)
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature
- 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:

Sampling method: Samples for possible analysis were taken from all test concentrations and the control at t=0 and t=48h. 1.8 mL of volume was taken from the approximate centre of the test vessels. Samples were stored in a freezer (≤-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.

Additionally, reserve samples of 1.8 mL were taken for possible analysis.

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 the test medium. Thereafter, the aqueous Saturated Solution (SS) was collected by filtration through a 0.45 µm membrane filter (RC55, Whatman) and 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 test solutions were 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:

no

Total exposure duration:

48 h

Test conditions

Hardness:

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

Test temperature:

20-21°C (requirement: 18-22°C, constant within ±1°C)

pH:

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

Dissolved oxygen:

At t=0: 8.4-8.5 mg/L
At t=48h: 9.0-9.1 mg/L

Salinity:

Not relevant

Nominal and measured concentrations:

Final test:
Nominal concentrations: 0 (blank), 0, 10, 18, 32, 56 and 100% of a SS prepared at 100 mg/L
Measured concentrations (mg/L) at t=0h: Measured concentrations (mg/L) at t=48h:
The geometric mean concentrations over the 48h test period, were 7.8, 14, 25, 43 and 78 mg/L at the loading rates of 10, 18, 32, 56 and 100% of the 100 mg/L SS, respectively.

LOQ is 0.1 mg/L
* Result of repeated analysis within the system stability on 09 Nov 2022 after storage on room temperature with minimal dilution since the response of initial analysis was outside the calibrated range

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 / WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium
- Culture medium different from test medium: Culture medium had extra trace elements, macro nutrients and vitamins added as specified in Medium M7.
OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: daily photoperiod of 16h
EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Immobility (including mortality), defined as an organism's inability to swim within 15 seconds after gentle agitation of the test container. This parameter was recorded at 24h and 48h.
RANGE-FINDING STUDY
- Test concentrations: 1.0, 10 and 100% of a SS prepared at 100 mg/L
- 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.:

44 mg/L

95% CI:

>= 38 - <= 51

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

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

The 24h-EC50 value was determined to be 62 mg/L (54-70 mg/L), based on geometric mean measured concentrations.

Results with reference substance (positive control):

No immobility was observed in the control throughout the test. At the end of the test, a dose-related increase of immobility was observed at 100% of the SS prepared at a loading rate of 100 mg/L reaching 100% immobility. Therefore, the expected EC50 was between 10 and 100 mg/L.

Reported statistics and error estimates:

The 24 and 48h-EC50-values were calculated from the probits of the percentages of affected daphnids and the logarithms of the corresponding test material concentrations (geometric mean) using the maximum likelihood estimation method.

ToxRat Professional (ToxRat Solutions®, GmbH, Germany) was used for the analysis.

Parameters of the Probit analysis at 24h:
Computation runs: 9
Slope b: 10.37143
Intercept a: -18.58707
Variance of b: 5.06098
Goodness of Fit Chi²: 0.00018
Degrees of freedom: 3.00000
p(Chi²): 1.00000
Log LC50: 1.79214
SE Log LC50: 0.02683
g-Criterion: 0.18074
F: 355636.06300
p(F) (df: 1;4): <0.001

Parameters of the Probit analysis at 48h:
Computation runs: 9
Slope b: 7.84576
Intercept a: -12.88668
Variance of b: 2.79160
Goodness of Fit Chi²: 1.56448
Degrees of freedom: 3.00000
p(Chi²): 0.66800
Log LC50: 1.64250
SE Log LC50: 0.02966
g-Criterion: 0.17421
F: 42.28300
p(F) (df: 1;4): 0.00700

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of JNJ-63632283-AAA (T003665) to Daphnia was determined in a 48 hour static test according to OECD guideline 202. The 48h-EC50 was 44 mg/L based on geometric mean concentrations (95% confidence interval between 38 and 51 mg/L).