Registration Dossier

Ecotoxicological information

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:
2016-04-04 to 2016-05-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study reports): JNJ-117676-AAA (T000263)
- Physical state: solid (powder)
- Appearance: White, slight beige amorphous, crystalline, micropowder
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: I15BB0491
- Expiration date of the lot/batch: 2021-02-04 (retest date)
- Purity test date: 2015-02-06

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature

OTHER SPECIFICS:
- Name of test material (as cited in study reports): JNJ-117676-AAA (T000263)
- Physical state: solid (powder)
- Appearance: white to slightly grey powder

specific details on test material used for the study:
- Analytical purity: 99.8% (base titration: >=98.0%)

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Sampling method: samples for possible analysis were taken form all test concentrations and the control at t=0 and t=48h. 3.0 mL of volume was taken from the approximate centre of the test vessels. At the end of the exposure period, the replicates were pooled at each concentration before sampling
- Sample storage conditions before analysis: the samples were stored in a freezer. Additionally, reserve samples of 3.0 mL were taken for possible analysis

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Preparation of test solutions started with the highest test concentration of 100 mg/L applying a 10-minute treatment period with ultrasonic waves followed by 17-26 minutes of magnetic stirring to accelerate the dissolution of the test item in test medium. After adjusting the pH from 9.5 (combined limit/range-finding test) and 9.6 (final test) to 8.5 with 1M HCl (Merck, Darmstadt, Germany), the 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): final test solutions were clear and colourless

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): <24h (from parental daphnids of more than 2 weeks old)
- Method of breeding: 250 newborn daphnids (i.e. less than 3 days old) are placed into 5 litres 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 test: no

ACCLIMATION
- Acclimation period: max. 4 weeks
- Acclimation conditions (same as test or not): yes
- Type and amount of food: fresh water algae.
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h

Test conditions

Hardness:
180 mg/L as CaCO3
Test temperature:
20-21°C
pH:
At t= 0h: 8.1 - 8.4
At t= 48h: 8.2
Dissolved oxygen:
At t= 0h: 8.7 - 9.3 mg/L
At t= 48h: 8.3 - 9.0 mg/L
Salinity:
not relevant
Nominal and measured concentrations:
Final test
Nominal concentrations: 0, 10, 18, 32, 56, 100 mg/L
Measured concentrations at t=0h: 0.00097, 10.7, 19.4, 34.6, 6.5, 109 mg/L
Measured concentrations at t=48h: 0.00097, 10.6, 19.2, 33.3, 60.3, 109 mg//L
Details on test conditions:
TEST SYSTEM
- Test vessel: beaker
- Material, size, headspace, fill volume: 100 mL, glass beaker filled with 80mL
- Aeration: not during the test
- Type of flow-through (e.g. peristaltic or proportional diluter): no flow-through
- Renewal rate of test solution (frequency/flow rate): no renewal, static system
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

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: x1.8
- Range finding study
- Test concentrations: 0.1, 1.0, 10 and 100 mg/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.:
42 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks on result:
other: 95% C.L.: 38 to 46 mg/L
Details on results:
- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: none
Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
Results with reference substance (positive control):
- Results with reference substance valid? yes
- EC50/LC50: 48h-EC50 = 0.29 mg/L (with a 95% C.L. interval between 0.26 and 0.31 mg/L). The sensitivity of the daphnia was within the range determined with the historical data collected at WIL Research Europe.
Reported statistics and error estimates:
The 48h-EC50 value was calculated from the probits of the percentages of affected daphnids and the logarithms of the corresponding nominal concentrations of the test item using the maximum likelihood estimation method.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of T000263 to Daphnia magna was determined in a 48-hour static test according to the OECD guideline 202.
Based on nominal test concentrations, the 48-h EC50 was determined to be 42 mg T000263/L (95% C.L. 38 to 46 mg/L). The results of the test can be considered reliable without restriction.