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Short-term toxicity to fish

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short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-09-25 to 2015-10-24
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
according to
OECD Guideline 203 (Fish, Acute Toxicity Test)
according to
EU Method C.1 (Acute Toxicity for Fish)
according to
other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, december 14, 2000
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
- Source and lot/batch No.of test material:
* Identification: JNJ-16479489-AAA (T001159)
* Batch: M14IB5308
* WIL Research test item number: 206568/B
- Expiration date of the lot/batch: 2015-11-21 (retest date)
- Purity test date: 2015-04-23

- Storage condition of test material: At room temperature
- Solubility and stability of the test substance in the solvent/vehicle: solubility in water: 0.26 g/L
Analytical monitoring:
Details on sampling:
- Sampling method: samples were taken from all test concentrations with surviving fish and the control before the start of the test and after 96 hours. 2.0 mL of volume was taken.
- Sample storage conditions before analysis: stored in a freezer until analysis. Addionally, reserve samples of 2.0 mL were taken for possible analysis.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: preparation of test solutions started with a loading rate of 100 mg/L applying 2 days of magnetic stirring in the dark to reach the maximum solubility of the test item in the test medium. The resulting aqueous mixture was filtered through a preconditioned 0.45 µm membrane filter (Whatman). The obtain filtrate (the saturated solution – SS) was used as the highest test concentration. The lower test concentrations were prepared by subsequent dilutions of the SS in test medium. The final test solutions were all clear and colourless.
- Controls: yes
- Evidence of undissolved material (e.g. precipitate, surface film, etc): final test solutions were clear and colourless
Test organisms (species):
Cyprinus carpio
Details on test organisms:
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands
- Length at study initiation (length definition, mean, range and SD): range-finding test: 2.2 ± 0.1 cm, final test: 1.9 ± 0.2 cm
- Weight at study initiation (mean and range, SD): range-finding test: 0.22 ± 0.02g, final test 0.24 ± 0.06 g
- Feeding during test: no

- Acclimation period: at least 12 days after delivery
- Acclimation conditions (same as test or not): yes
- Type and amount of food: pelleted fish food
- Feeding frequency: daily
- Health during acclimation (any mortality observed): mortality less than 5% 7 days before test
Test type:
Water media type:
Limit test:
Total exposure duration:
96 h
not reported
Test temperature:
Dissolved oxygen:
not relevant
not indicated
Nominal and measured concentrations:
- Range-finding test
* Nominal concentrations (% of a SS prepared at a loading rate of 100 mg/L): 10, 100
* Measured concentrations (mg/L) at t= 0h: 10.5, 142
* Measured concentrations (mg/L) at t= 18h: 10.8, 104
* Measured concentrations (mg/L) at t= 96h: 10.7, 105

- Final test
* Nominal concentrations (% of a SS prepared at a loading rate of 100 mg/L): 0, 32, 56, 100
* Measured concentrations (mg/L) at t= 0h: 0.00021, 37.4, 66.0, 119
* Corrected concentration (mg/L) at t= 0h: 0.00016, 29.8, 52.7, 94.9
* Measured concentrations (mg/L) at t= 18h: x, x, x, 121
* Corrected concentration (mg/L) at t= 18h: x, x, x, 96.2
* Measured concentrations (mg/L) at t= 96h: 0.00026, 38.4, 67.3, x
* Corrected concentration (mg/L) at t= 96h: 0.00020, 30.6, 53.6, x
Details on test conditions:
- Test vessel: beaker
- Material, size, headspace, fill volume: 6L, all-glass filled with 5L test solution
- Aeration: not during the test
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): not reported
- No. of vessels per control (replicates): not reported
- Biomass loading rate: 0.34 g fish/L

- Adjustment of pH: 7.7 ± 0.3
- Photoperiod: 16h light daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Fish were considered to be dead when no reaction was observed after touching the caudal peduncle and visible breathing movements were absent. In addition, fish that were convulsing or showing other severe forms of distress not considered transient in nature and likely to become more severe before the exposure is terminated, will be sacrificed for humane reasons. These fish will be treated as having died in the test, based on OECD guideline on humane endpoint (ENV/JM/MONO/(2000)7).

- Spacing factor for test concentrations: x1.8
- Range finding study: combined limit/range finding study was performed
- Test concentrations: 10, 18, 32, 56 and 100 % of a SS prepared at a loading rate of 100 mg/L
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
Key result
96 h
Dose descriptor:
Effect conc.:
55 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Remarks on result:
other: 95% C.L.: 44-72 mg/L
Details on results:
- Mortality of control: none
- Other adverse effects control: no
- Abnormal responses: none
Results with reference substance (positive control):
- Results with reference substance valid? yes
- EC50/LC50: 96h-LC50 = 0.32 mg/L.
Reported statistics and error estimates:
The LC50-values were calculated from the weibits of the percentages of affected fish and the logarithms of the corresponding nominal test item concentrations using linear maximum likelihood regression.
Validity criteria fulfilled:
The acute toxicity of T001159 to Carp was determined in a 96-hour static test according to the OECD guideline 203.
Under the conditions of the present study, The 96-h LC50 was 55 mg/L (95% confidence interval between 44 and 72 mg/L) based on analytically confirmed nominal concentrations and was already reached after 48 hours of exposure. The results of the test can be considered reliable without restriction.

Description of key information

The study of Bouwman (2016), investigating the acute toxicity of T001159

in carp according to OECD guideline 203, was considered as the key study for endpoint coverage. An LC50 of 55 mg/L was found after an exposure of 96 hours in a static test in freshwater. Based on the results it can be considered that T001159 is harmful to fish.

Key value for chemical safety assessment

Additional information

One key study of Bouwman (2016) was available on the acute toxicity of T001159 to carp. Carp was exposed for 96 hours to T001159 under static conditions. The nominal test item concentrations were 10, 18, 32, 56 and 100 mg/L and in parallel the control. In the analyzed test samples from the start of the test medium renewal periods the measured test item concentrations ranged between 88 and 100% of the nominal values. As the mean measured concentrations showed that the test levels have been satisfactorily maintained over the exposure period (ie. >80% of the nominal concentration), the nominal concentrations were used to express toxicity. Based on the nominal test concentrations a 96 -h LC50 of 55 mg/L for T001159 (with 95% confidence limits of 44 and 72 mg/L) was obtained.

Based on the results of this study, it can be considered that T001159 is harmful to fish.