1-[2-[2-methoxy-4-(morpholin-4-yl)-5-nitroanilino]pyrimidin-4-yl]-3-phenyl-1H-pyrazole-4-carbaldehyde

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:

12 May 2021 to 21 Oct 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2022

Report date:

2022

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

April 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

Version / remarks:

February 2019

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: Guidance Document on the Use of the Harmonised System for the Classification of Chemicals which are Hazardous for the Aquatic Environment, OECD series on testing and assessment number 27

Version / remarks:

July 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1-[2-[2-methoxy-4-(morpholin-4-yl)-5-nitroanilino]pyrimidin-4-yl]-3-phenyl-1H-pyrazole-4-carbaldehyde

Cas Number:

1903010-39-8

Molecular formula:

C25H23N7O5

IUPAC Name:

1-[2-[2-methoxy-4-(morpholin-4-yl)-5-nitroanilino]pyrimidin-4-yl]-3-phenyl-1H-pyrazole-4-carbaldehyde

Test material form:

solid: particulate/powder

Details on test material:

- Physical state: solid
- Appearance: orange powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch (Lot) Number: I20ED2016
- Purity/Composition: 101.5%
- Purity test date : 2020-07-27
- Expiry date: 2022-06-06 (retest date)
- Physical Description: Orange powder

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 item in the solvent/vehicle: not indicated

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 from the approximate centre of the test vessels. 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:

yes

Remarks:

acetone

Details on test solutions:

Preparation of Test Solutions
Method: the batch tested was an orange powder with a purity of 101.5% and was visually completely soluble in test medium at the concentrations tested. No correction was made for the purity/composition of the test item. Due to the low solubility of the test item in water, stock solutions were prepared in acetone.
Preparation of stock solutions, i.e. test item in acetone, started with the highest concentration of 0.010 mg/mL. No other treatment than vigorous shaking was needed to completely dissolve the test item in acetone. Lower stock concentrations, i.e. 0.0010 and 0.00010 mg/mL, were prepared by subsequent dilutions of the highest concentration in acetone.
Subsequently, 100 μL of each stock solution were added to respectively 1.0 L test medium, whilst stirring, to obtain the according final test concentrations. The mixtures were magnetically stirred for a period of approximately 15 minutes to ensure a homogenous distribution of the test item in the test solutions. For the solvent control, 100 μL acetone was added to 1.0 L test medium, whilst stirring.
- Evidence of undissolved material: 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
- Straus, 1820; at least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions.
- 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, constant within ±1°C

pH:

At t=0h: 7.5 - 7.6
At t=48h: 8.1 - 8.2

Dissolved oxygen:

At t-0h: 8.2 - 8.8
At t-48h: 8.6 - 8.7

Salinity:

Not relevant

Nominal and measured concentrations:

combined limit/range-finding test=
- nominal: 0.010, 0.10, 1.0 µg/L
- controls: blank-control and solvent-control
- measured concentrations (for 1.0 µg/L): t=0h: 1.44, t=48h: 0.50 (below LOQ of 1.0 µg/L, hence half of the LOQ was used). Geometric mean: 0.85 µg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 60 mL, all glass
- Material, size, headspace, fill volume: filled with 50 mL of test solution
- Aeration: not applied
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for highest concentration, 2 for intermediate concentrations
- 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 (even if they can still move their antennae) and was monitored after 24 and 48 hours of continuous treatment with the test substance.

TEST CONCENTRATIONS
- combined range-finder/limit test
- Test concentrations: 0.010, 0.10, 1.0 µg/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.:

> 0.85 µg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Details on results:

- Immobility of control: none
- Other adverse effects control: no
- Abnormal responses: none
- EC50 (24h) > 0.85 µg/L

Results with reference substance (positive control):

The batch of Daphnia magna tested showed expected sensitivity to Potassium dichromate (Batch K50664264, Merck KGaA, Darmstadt, Germany), based on the historical range of reference tests performed by the Test Facility in the last ten years.

Reported statistics and error estimates:

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

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of T003902 to Daphnia was determined in a 48 hour static test according to OECD guideline 202. In conclusion, the 48h-EC50 for Daphnia magna exposed to JNJ-73848255-AAA (T003902) exceeded the maximum solubility of the test item in test medium, i.e. exceeded a geometric mean concentration of 0.85 µg/L. The results of the test can be considered reliable without restriction.