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Ecotoxicological information

Short-term toxicity to aquatic invertebrates

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short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-10-06 to 2017-11-29
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
according to guideline
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
according to guideline
EU Method C.2 (Acute Toxicity for Daphnia)
according to guideline
other: Guidance document on aquatic toxicity testing of difficult items and mixtures, OECD series on testing and assessment number 23, December 14, 2000
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
- Source and lot/batch No.of test material: M16LB4523
- Expiration date of the lot/batch:2018-02-09 (retest date)
- Purity test date: 2017-04-19 (certificate of analysis release date)

- Storage condition of test material: at room temperature
- Stability under test conditions: not indicated
- Solubility and stability of the test substance in the solvent/vehicle: not indicated
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. 2.0 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 2.0 mL were taken for possible analysis.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to an expected low water solubility of the test item, saturated solutions (SS) were prepared. The preparation of test solutions started with a loading rate of 100 mg/L applying three days of magnetic stirring at room temperature to ensure maximum dissolution of the test item in test medium and full degradation of the test item to its metabolite. This resulted in a clear and colourless dispersion that contained undissolved material. The obtained mixture was filtered through a 0.45 µm membrane filter (Whatman; RC55) to remove the undissolved fraction. pH of the obtained SS was set from 4.3 to 6.8 in the combined limit/range-finding test, from 4.4 to 6.8 in the first full test and from 4.5 to 8.2 in the limit test with 1 M NaOH (Merck, Darmstadt, Germany) and used as the highest test concentration. Lower test concentrations were prepared by subsequent dilutions of the SS in test medium.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): final test solutions were clear and colourless
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): < 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 period: not relevant
Test type:
Water media type:
Limit test:
Total exposure duration:
48 h
180 mg/L as CaCO3
Test temperature:
At t=0h: 8.0-8.1
At t=48h: 8.0 - 8.2
Dissolved oxygen:
At t=0h: 9.0-9.5
At t=48h: 9.3-9.4
not relevant
Nominal and measured concentrations:
Range finder:
Nominal concentrations: 1.00, 10 and 100% of SS (1% of SS not analysed)
Measured concentrations (mg/L) at t=0h: 9.59, 88.8
Measured concentrations (mg/L) at t=48h: 9.60, 88.0

Final test 1:
Nominal concentrations: Saturated solution prepared at the regulatory limit concentration of 100 mg/L
Measured concentrations (mg/L) at t=0h: 0.027, 89.1
Measured concentrations (mg/L) at t=48h: n.d., 87.1

Final test 2:
Nominal concentrations: Saturated solution prepared at the regulatory limit concentration of 100 mg/L
Measured concentrations (mg/L) at t=0h: n.d., 102
Measured concentrations (mg/L) at t=48h: n.d., 92.1
Details on test conditions:
- 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
- No. of vessels per control (replicates): 4
- Test medium: adjusted ISO medium

- 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: x100
- Range finding study
- Test concentrations: 1, 10 and 100% of a SS prepared at a loading rate of 100 mg/L (setup: combined range finder/limit test)
- Results used to determine the conditions for the definitive study: yes
Reference substance (positive control):
Potassium dichromate
Key result
48 h
Dose descriptor:
Effect conc.:
> 100 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
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
- Mortality: no
- EC50/LC50: 48-h EC50 = 0.41 mg/L. The sensitivity of the daphnids was within the range determined with the historical data collected at Charles River Laboratories Den Bosch.
Reported statistics and error estimates:
The 24 / 48-h EC50 could not be calculated because the observed effects were below 50%
Validity criteria fulfilled:
The acute toxicity of 4-nitrobenzene-1-sulfonylchloride to Daphnia magna was determined in a 48 hour static test according to OECD guideline 202. The 48-h EC50 exceeded the analytically confirmed nominal concentration of 100 mg/L. The results of the test can be considered reliable without restriction.

Description of key information

The study of Tobor-Kaplon (2018), investigating the acute toxicity of 4-nitrobenzene-1-sulfonylchloride in Daphnia magna according to OECD guideline 202, was considered as the key study for endpoint coverage. An EC50 >100 mg/L (based on analytically confirmed nominal test concentrations) was found after an exposure of 48 hours in a static test in freshwater. Based on the results it can be concluded that 4-nitrobenzene-1-sulfonylchloride is not harmful to aquatic invertebrates.

Key value for chemical safety assessment

Additional information