tert-Butyl 4-({6-[7-cyclopentyl-6-(dimethylcarbamoyl)-7H-pyrrolo[2,3-d]pyrimidin-2-yl]amino}pyridin-3-yl)piperazine-1-carboxylate

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Feb - June 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Singular samples for possible analysis were taken from all test concentrations and the control according to the schedule.
Frequency at t=0 h and t=48 h
Volume 2.0 ml from the approximate centre of the test vessels
Storage Samples were stored in a freezer until analysis.
At the end of the exposure period, the replicates were pooled at each concentration before sampling.

Details on test solutions:

LEE011-A3 Solutions containing 1.0, 10 and 100% of the WSF prepared at a loading rate 100 mg/l.
Controls Test medium without test substance or other additives.

The standard test procedures required generation of test solutions, which should contain completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that disturb the test system should be prevented (e.g. film of the test
substance on the water surface). No correction was made for the purity/composition of the test substance.
The batch of LEE011-A3 tested was a beige powder and was not completely soluble in test medium at the loading rate initially prepared.
Preparation of test solutions started with a loading rate of 100 mg/l applying a 2-day period of magnetic stirring followed by a filtration through a 0.45 μm membrane filter (rc55, Whatman). The obtained Water Soluble Fraction (WSF) was used as the highest test concentration. The lower test
concentrations were prepared by subsequent dilutions of the WSF in test medium. The final test solutions were all clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

18-22°C

pH:

6.0-8.5

Dissolved oxygen:

>=3 mg/l

Nominal and measured concentrations:

The actual test concentration at the start of the test was 0.24 mg/l for the WSF prepared at 100 mg/l and the concentration remained stable during the test (98% initial). This concentration was considered to be maximum soluble concentration in test medium.

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.24 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks:

and mortality

Results with reference substance (positive control):

The reference test was carried out to check the sensitivity of the test system as used by WIL Research Europe. Daphnia were exposed for a maximum of 48 hours to K2Cr2O7 concentrations of 0.10, 0.18, 0.32, 0.56, 1.0 and 1.8 mg/l and to a control. Twenty daphnids were exposed per concentration.
The actual responses in this reference test with K2Cr2O7 are within the ranges of the expected
responses at the different concentrations, i.e. the 48h-EC50 was between 0.3 and 1.0 mg/l. Hence, the
daphnia were rather sensitive and the 48h-EC50 was just within the range determined with the
historical data collected at WIL Research Europe.
The 24h-EC50 was 0.60 mg/l with a 95% confidence interval between 0.52 and 0.69 mg/l.
The 48h-EC50 was 0.29 mg/l with a 95% confidence interval between 0.25 and 0.34 mg/l.

Validity criteria fulfilled:

yes

Conclusions:

The 48h-EC50 was beyond the range tested, e.g. exceeded the maximum soluble concentration in the test medium, i.e. 48h-EC50 >0.24 mg/l.

Description of key information

The 48h-EC50 was beyond the range tested, e.g. exceeded the maximum soluble concentration in the test medium, i.e. 48h-EC50 >0.24 mg/l.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.24 mg/L