2-Pyridinecarbonitrile, 5-amino-3-(trifluoromethyl)-

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 Jan 2021 to 26 Feb 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

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

5-amino-3-(trifluoromethyl)pyridine-2-carbonitrile

EC Number:

815-137-9

Cas Number:

573762-62-6

Molecular formula:

C7H4F3N3

IUPAC Name:

5-amino-3-(trifluoromethyl)pyridine-2-carbonitrile

Details on test material:

- Appearence : Slightly yellowish brown fine powder
- CAS Number: 573762-62-6
- Molecular formula: C7H4F3N3
- Molecular weight: 187.12

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- lot/batch number of test material: BI03K2005005
- Expiry date: 02 June 2021 (retest date)
- Physical Description: Almost white to yellow powder (determined by Charles River Den Bosch)
- Purity: 99.7%
- Purity correction factor: no correction factor required
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. 2.0 mL of volume was taken from the approximate centre of the test vessels. Samples were stored in a freezer (≤-15°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 the highest concentration of 100 mg/L applying one hour of ultrasonic treatment to accelerate dissolution of the test item in medium. lower test concentrations were prepared by subsequent dilutions of the highest concentration 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:

18-22°C, constant within ±1°C

pH:

At t=0h: 7.6 - 7.7
At t=48h: 8.0

Dissolved oxygen:

At t-0h: 9.4 - 9.9 mg/L
At t-48h: 9.0 - 9.6 mg/L

Salinity:

Not relevant

Nominal and measured concentrations:

final test:
- nominal concentrations: 0 (control),10, 18, 32, 56 and 100 mg/L
- measured concentrations: t-0h: nd, 9.30, 18.3, 31.8, 58.8, 102 mg/L
- measured concentrations: t-48h: nd, 9.91, 17.7, 31.8, 60.1, 98.3 mg/L

The measured concentrations were in agreement with the nominal concentrations, i.e. varied between 93 and 107% relative to the nominal values at the start and the end of the 48-hour exposure period. No test item was detected in the samples taken from the control solution. Based on these results, effect parameters were expressed as analytically confirmed nominal concentrations

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 (range-finding study)
- Spacing factor for test concentrations: 10
- Range finding study
- Test concentrations: 0.10, 1.0 and 10 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.:

79 mg/L

95% CI:

>= 73 - <= 86

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

not specified

Details on results:

- Immobility of control: none
- Other adverse effects control: no
- Abnormal responses: none
- EC50-24h: 79 mg/L

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: no
- Limit test:
- EC50/LC50: 48-h EC50 = 0.48 mg/L. The sensitivity of the daphnids was within the range determined with the historical data collected at Charles River Laboratoire Den Bosch.

Reported statistics and error estimates:

The EC50 could not be determined using a regression method. Instead, the 24 and 48h-EC50-values were calculated by applying the Spearman-Karber procedure (non-linear; without trimming) on the percentages of affected daphnids and the logarithms of the corresponding nominal test item concentrations.ToxRat Professional v 3.2.1 (ToxRat Solutions® GmbH, Germany) was used to perform the analysis

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of T003630 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-61232574-AAA (T003630) was 79 mg/L based on analytically confirmed nominal concentrations (95% confidence interval between 73 and 86 mg/L). The results of the test can be considered reliable without restriction.