1-[(tert-butoxycarbonyl)amino]cyclobutane-1-carboxylic acid

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:

27 Oct 2020 to 18 Dec 2020

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:

1-[(tert-butoxycarbonyl)amino]cyclobutane-1-carboxylic acid

EC Number:

627-199-2

Cas Number:

120728-10-1

Molecular formula:

C10H17NO4

IUPAC Name:

1-[(tert-butoxycarbonyl)amino]cyclobutane-1-carboxylic acid

Test material form:

solid: particulate/powder

Details on test material:

Physical state: powder
Appearance: white powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- lot/batch number of test material: AC07N1912008
- Expiry date: 02 January 2021 (retest date)
- Physical Description: Almost white powder
- Purity: 98.9%
- Purity correction factor: not 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 a twenty-five-minute period of ultrasonic waves 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:

20-21°C

pH:

At t=0h: 6.4* - 7.7
At t=48h: 7.7 - 8.0
*The pH 6.4, measured at the start of the test for test concentration 100 mg/L, was within the range of 6 to 9. There was not more than 1.5 units difference with the pH of test concentration 100 mg/L measured at the end of the test and also not between all test concentrations at the start of the test. There was 1.6 units difference between this pH 6.4 and the pH measured at the end of the test for the control and test concentration 0.10 mg/L. However, as no immobility was observed in the control and at any of the test concentrations throughout the exposure period, this variance in pH was assumed to have no effect on the validity and integrity of the test.

Dissolved oxygen:

At t-0h: 8.4 - 8.8
At t-48h: 7.7 - 8.0

Salinity:

Not relevant

Nominal and measured concentrations:

nominal: 0.10, 1.0, 10 and 100 mg/L

Samples taken from the control and the highest test concentration were analysed. The analysed concentrations of the first batch of samples were considered not reliable, since the mean recovery of the high QC sample was outside the criterion of 70-110%, i.e. 123%.
The measured concentration in the reserve sample of the highest test concentration taken at the start of the test was 107 mg/L. At the end of the exposure period, the measured concentration was at 97% relative to the initial concentration.

Based on these results, effect parameters were expressed as analytically confirmed nominal concentrations.

Please refer to the tables below under 'Any other information on materials and methods incl. tables'

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 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 and was monitored after 24 and 48 hours of continuous treatment with the test substance.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Range finding study
- Test concentrations: 0.10, 1.0, 10 and 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.:

> 100 mg/L

Nominal / measured:

nominal

Details on results:

- Immobility of control: none
- Other adverse effects control: no
- Abnormal responses: none
- 24h-EC50: > 100 mg/L

Results with reference substance (positive control):

- Results with reference substance valid? yes
- The batch of Daphnia magna tested, showed expected sensitivity to Potassium dichromate based on the range specified in ISO International Standard 6341, October 2012

Reported statistics and error estimates:

No EC50 could be calculated because the test item proved to be non-toxic (EC50 > maximum concentration tested)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Toxicity of T003661 to Daphnia magna was tested according to OECD 202 guideline using test concentrations of 0.10, 1.0, 10 and 100 mg/L during 48 hours. In conclusion, the 48h-EC50 for Daphnia magna exposed to JNJ-63543883-AAA (T003661) was beyond the range of concentrations tested, i.e. EC50 > 100 mg/L..