4-chloro-N-[2-methoxy-4-(morpholin-4-yl)-5-nitrophenyl]pyrimidin-2-amine

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:

23 APR 2021 to 10 MAY 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-chloro-N-[2-methoxy-4-(morpholin-4-yl)-5-nitrophenyl]pyrimidin-2-amine

Cas Number:

2270944-70-0

Molecular formula:

C15H16ClN5O4

IUPAC Name:

4-chloro-N-[2-methoxy-4-(morpholin-4-yl)-5-nitrophenyl]pyrimidin-2-amine

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: I20CD1048
- Expiration date of the lot/batch: 2022-0-22 (retest date)
- Purity test date: 2020-05-11 (certificate of analysis release date)
- Physical appearance: Light yellow to dark orange solid

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

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. 1.5 mL of volume was taken from the approximate centre of the test solutions. 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:

The batch of JNJ-73848242-AAA (T003901) tested was a light yellow to dark orange solid with a purity of 100.7% and was not completely soluble in test medium at the loading rate initially prepared. No correction was made for the purity/composition of the test item.
Preparation of test solutions started with a loading rate of 100 mg/L applying fifteen-minutes of ultrasonic waves followed by a three-day period of magnetic stirring to ensure maximum
dissolution of the test item 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 highest test concentration. Lower test concentrations were prepared by subsequent dilutions of the SS in test medium. 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: < 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: suspension 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

Test temperature:

19-21 °C

pH:

control (start/end): 7.8/8.1, 100 mg/L (start/end): 7.8/8.1

Dissolved oxygen:

control (start/end): 8.5/9.2 mg/L, 100 mg/L (start/end): 8.5/9.1

Nominal and measured concentrations:

Nominal: Solutions containing 0.10, 1.0, 10, and 100% of the SS prepared at a loading rate of 100 mg/L, measured: Highest test concentration: 0.39 mg/L (Geometric mean concentration). //

Combined limit/range finding test: Samples taken from the highest test concentration were analysed. The measured concentration at the start of the test was 0.39 mg/L. During the exposure period, the concentration remained stable, i.e. was at 100% of initial at the end of the test. Based on these results, the geometric mean concentration was calculated to be 0.39 mg/L for the 100% SS concentration, this was used to express effect parameters.

Details on test conditions:

TEST SYSTEM
- Test vessel: beaker, 60 mL, all-glass
- Aeration: not during the test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for highest, intermediate: 2
- No. of vessels per control (replicates): 4
- Test medium: The following salts (analytical grade) were added to tap water purified by Reverse Osmosis: CaCl2.2H2O: 211.5 mg/L, MgSO4.7H2O: 88.8 mg/L, NaHCO3: 46.7 mg/L, KCl: 4.2 mg/L

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: 10
- combined Limit/Range finding study
- Test concentrations: 0.10, 1.0, 10 and 100% of a SS prepared at 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.:

> 0.39 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- EC50 (24h): > 0.39 mg/L
- Immobility of control: none
- Other adverse effects control: no
- Abnormal responses: none

- Effect concentrations exceeding solubility of substance in test medium: yes

Results with reference substance (positive control):

expected sensitivity of test organism towards potassium dichromate

Reported statistics and error estimates:

No EC50 could be calculated because the test item proved to be not acutely toxic to Daphnia magna (EC50 > maximum concentration tested)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The objective of the study was to evaluate JNJ-73848242-AAA (T003901) for its ability to generate acute toxic effects on the mobility of Daphnia magna during an exposure period of 48 hours and, if possible, to determine the EC50 at 24 and 48 hours of exposure. A combined limit/range-finding test and saturation test were performed. The study met the acceptability criteria prescribed by the study plan and was considered valid. In conclusion, under the conditions of the present study with Daphnia magna, no immobility or other effects were recorded at any of the concentrations of JNJ-73848242-AAA (T003901) tested. The 48h-EC50 for Daphnia magna exposed to JNJ-73848242-AAA (T003901) was beyond the range of concentrations tested, i.e. exceeded a geometric mean measured concentration of 0.39 mg/L, which was considered the maximum soluble concentration in test medium. The results of the test can be considered reliable without restriction.