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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

Administrative data

short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-01-13 to 2017-02-20
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report Date:

Materials and methods

Test guidelineopen allclose all
according to
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
according to
EU Method C.2 (Acute Toxicity for Daphnia)
GLP compliance:
yes (incl. certificate)

Test material

Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study reports): T000836 (JNJ-16250351-AAH)
- Physical state: solid
- Appearance: white powder
Specific details on test material used for the study:
- Source and lot/batch No.of test material: I16DB2126
- Expiration date of the lot/batch: 2018-04-27 (retest date)
- Purity test date: 2016-05-27

- Storage condition of test material: At room temerature
- Stability under test conditions: not indicated
- Solubility and stability of the test substance in the solvent/vehicle: solubility in water: >500 g/L

Sampling and analysis

Analytical monitoring:
Details on sampling:
- Sampling method: samples for possible analysis were taken from all test concentrations and the control at t=0 and t=48h. 2.4 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. Addionally, reserve samples of 2.4 mL were taken for possible analysis

Test solutions

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 130 mg/L (100 mg base/L) applying 1 hour of magnetic stirring at room temperature to ensure maximum dissolution of the test item in test medium. This resulted in a clear and colourless solution with a few very finely dispersed undissolved particles that were not considered as disruptive to the test system and therefore not removed. The resulting solution was used as the highest test concentration. Lower test concentrations were prepared by subsequent dilutions 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:
- 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: M7. After 7 days of cultivation half of the medium is renewed twice a week.
- Feeding during test: no

- Acclimation period: not relevant

Study design

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

Test conditions

180 mg/L CaCO3
Test temperature:
19.5 - 21.6 °C
At t= 0h: 7.9 - 8.0
At t= 48h: 7.8 - 7.9
Dissolved oxygen:
At t= 0h: 9.5 - 9.9
At t= 48h: 9.1 - 9.7
not relevant
Nominal and measured concentrations:
Range finder:
Nominal concentrations (mg/L): 0.13, 1.3, 13 and 130 (0.10, 1.0, 10 and 100 mg base/L)

Final test
Nominal concentrations tested (mg/L): 6.0, 13.0, 28.6, 59.8 and 130 (4.6, 10, 22, 46 and 100 mg base/L)
Measured base concentrations (mg/L) at t=0h: 4.6, 10.5, 23.2, 48.2 and 101
Measured base concentrations (mg/L) at t=48h: 3.35, 7.70, 17.2, 35.2 and 74.6

Measured concentrations were in agreement (100-106%) with nominal at the study start. Measured concentrations decreased to 73-74% of initial at the end of the test. Based on these results, the average exposure concentrations were calculated. Average base concentration: 3.9, 9.0, 20, 41 and 87 mg base/L. Average test item concentration: 5.1, 12, 26, 53 and 113 mg/L.
Details on test conditions:
- Test vessel: beaker
- Material, size, headspace, fill volume: 100 mL, glass beaker filled with 80mL
- Aeration: not during the test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): four replicates
- No. of vessels per control (replicates): four replicates

- 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.

- Spacing factor for test concentrations: x 2.17
- Range finding study: combined limit/range finding study was performed
- Test concentrations:
Limit/range: 0.13, 1.3, 13 and 130 (0.10, 1.0, 10 and 100 mg base/L)
Final test: 6.0, 13.0, 28.6, 59.8 and 130 (4.6, 10, 22, 46 and 100 mg base/L)
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
potassium dichromate

Results and discussion

Effect concentrations
Key result
48 h
Dose descriptor:
Effect conc.:
11 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
Details on results:
- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: none
- 48h-EC50 = 11 mg/L based on average exposure concentrations. This corresponds with 8.5 mg base/L.
Results with reference substance (positive control):
- Results with reference substance valid? yes
- 24h-EC50 = 0.91 mg/L (with a 95% C.L. between 0.77 and 1.1 mg/L)
- 48h-EC50 = 0.58 mg/L (with a 95% C.L. between 0.49 and 0.67 mg/L)
Th 48h-EC50 was within the expected range of 0.28 to 0.9 mg/L
The sensitivity of the daphnia was within the range determined with the historical data collected at Charles River Den Bosch.
Reported statistics and error estimates:
Average exposure concentrations were calculated as the geometric means of the concentrations of JNJ-16250351-AAH (T000836) measured in the samples taken at the start (t=0) and at the end (t=48h) of the test.
- The 24h-EC50 was calculated from the geometric mean of the two concentrations with no and complete effects, respectively.
- The 48h-EC50-value was calculated from the weibits of the percentages of affected daphnids and the logarithms of the corresponding average measured test item concentrations using the maximum likelihood estimation method.
The 48h calculations were performed with ToxRat Professional v. 3.2.1. (ToxRat Solutions® GmbH, Germany).

Applicant's summary and conclusion

Validity criteria fulfilled:
The acute toxicity of JNJ-16250351-AAH (T000836) to Daphnia magna was determined in a 48-hour static test, according to the OECD guideline 202.
Analyses showed that measured concentrations (based on the base content) were in agreement with nominal at the study start (100-106% relative to nominal). Measured concentrations decreased to 73-74% of initial at the end of the test. Based on these results, the average exposure concentrations were calculated. The range tested based on average measured concentrations was 3.9, 9.0, 20, 41 and 87 mg base/L. This corresponded with 5.1, 12, 26, 53 and 113 mg/L based on the whole test item.
The 48h-EC50 was 11 mg/L based on average exposure concentrations (95% confidence interval between 9.1 and 12 mg/L). This corresponded with 8.5 mg base/L (95% confidence interval between 7.0 and 9.2 mg base/L). The results of the test can be considered reliable without restriction.