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EC number: 229-175-3 | CAS number: 6422-83-9
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- Endpoint summary
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- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
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- Toxicological Summary
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Short-term toxicity to aquatic invertebrates
Administrative data
Link to relevant study record(s)
- Endpoint:
- short-term toxicity to aquatic invertebrates
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2003-04-23 to 2003-09-01
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.2 (Acute Toxicity for Daphnia)
- Version / remarks:
- December 1992
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
- Version / remarks:
- adopted April 4, 1984
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- yes
- Details on sampling:
- Frequency: One day before the start from the highest and lowest test group (check test system), at the start and the end of the exposure period from all test concentrations.
Volume: 10 ml from the approximate centre of the test vessels.
Storage: Not applicable, samples were analysed on the day of sampling.
- Vehicle:
- yes
- Remarks:
- DMSO
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solutions were prepared in DMSO at a factor of 10,000 higher than the target concentrations and dosed via a computer-controlled system consisting of 5 dispensers. Via this system the dosed volumes entered mixing flasks separately from the tap-water supply. The tap-water was supplied via a flow-meter with a flow rate of ca. 6 L/h.
- Controls: Tap-water without test substance but with the additive used in the treatment of the stocks, i.e. DMSO.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Dimethylsulfoxide (DMSO) - Test organisms (species):
- Daphnia magna
- Details on test organisms:
- TEST ORGANISM
- Common name: water flea
- Strain/clone: Daphnia magna (Crustacea, Cladocera) (Straus, 1820)
- Age of parental stock (mean and range, SD): Daphnids originated from a healthy stock, 2nd to 5th brood
- Feeding during test: No
- Food type: During breeding daily, a suspension of fresh water algae.
ACCLIMATION
- Acclimation period: With new-born animals, i.e. less than 3 days old, by placing about 250 of them into 10 litres of medium in an all-glass culture vessel.
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: fresh water algae
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no mortality > 20% observed
- Test type:
- flow-through
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 48 h
- Hardness:
- 250 mg/L CaCO3
- Test temperature:
- 18-22°C, constant within 2°C.
- pH:
- Between 6.0 and 8.5. Should not vary by more than 1 unit at the end of the test in any test solution.
- Dissolved oxygen:
- > 7 mg/l at the start of the test,
>= 5 mg/l at the end of the test. - Nominal and measured concentrations:
- nominal concentrations:
0, 0.1, 1, 10, 100 mg/L for the combined limit/range-finding test (static)
0, 0.1, 1.0, 10 mg/L for the additional range-finding test (flow-through)
0, 0.10, 0.22, 0.46, 1.0, 2.2 mg/L for the final flow-through study
Measured conentrations:
0, 0.035, 0.4, 5.0 mg/L for the additional range-finding test (flow-through)
0, 0.059, 0.085, 0.19, 0.31, 0.82 mg/L for the final flow-through study (mean measured) - Details on test conditions:
- TEST SYSTEM
- Test vessel:Stock solutions were prepared in DMSO at a factor of 10,000 higher than the target concentrations. The stocks were dosed via a computer-controlled system consisting of 5 dispensers (Gilson). Via this system the dosed volume entered a mixing flask separately from the tap-water supply. The tap-water was supplied via a flow meter with a flow rate of ca. 6 L/h. In the mixing flask the dosed volume and the tap-water were mixed under continuous stirring. The test system was checked daily.
- Material, size, headspace, fill volume: Stainless steel vessels with a volume of 1.5 litres containing 4 mesh containers of stainless steel.
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): The tap-water was supplied via a flow meter with a flow rate of ca. 6 I/h.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap Water
- Metals:
Selenium < 0.1 µg/L
Arsenic< 5.0 µg/L
Cadmium< 0.4 µg/L
Chromium <1.0 µg/L
Copper 65/32 µg/L
Mercury < 0.05 µg/L
Lead < 5.0 µg/L
Zinc 24/62 mg/L
Manganese < 0.010 mg/L
Calcium 12 mg/L
Magnesium 1.4 mg/L
Iron 0.061 mg/L
- Culture medium different from test medium: Medium used for culturing: M7, as prescribed by Dr. Elendt-Schneider (Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).
OTHER TEST CONDITIONS
- Photoperiod: 16 h daily
- Light intensity: 671 lux
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mobility
VEHICLE CONTROL PERFORMED: yes
RANGE-FINDING STUDY
- Test concentrations: nominal:
0, 0.1, 1, 10, 100 mg/L for the combined limit/range-finding test (static)
0, 0.1, 1.0, 10 mg/L for the additional range-finding test (flow-through)
- Results used to determine the conditions for the definitive study:
All daphnids exposed to the target concentration 10 mg/L became immobilised at the end of the test period, while 70% immobility was observed in the 1.0 mg/L target concentration. No immobility was observed in the 0.10 mg/L target concentration and the control. Analytical results showed that measured concentrations were at 30 to 60% of the target concentrations. The relatively low recoveries were not expected as the test solutions appeared to be completely dissolved and continuous renewal should avoid decrease of test concentrations by degradation. Analyses of a three-day old stock solution of 10 g/L used to dose the 1.0 mg/L target concentration showed that the stock was prepared correctly and was stable for at least three days. Consequently, it was decided to prepare stock solutions only once during the final test and to reduce the flow of the dilution water from 12 L/h to 6 L/h to prepare test solutions with actual exposure concentrations that were closer to the target concentrations. - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
- Duration:
- 24 h
- Dose descriptor:
- EC50
- Effect conc.:
- 712.5 µg/L
- Nominal / measured:
- meas. (arithm. mean)
- Remarks on result:
- other: 603.6 - 938.6 µg/l
- Duration:
- 48 h
- Dose descriptor:
- EC50
- Effect conc.:
- 591.4 µg/L
- Nominal / measured:
- meas. (arithm. mean)
- Remarks on result:
- other: 524.5 - 715.8 µg/l
- Results with reference substance (positive control):
- - Results with reference substance valid?
- Relevant effect levels:
The 24h-EC50 was 1.2 mg/L with a 95% confidence interval between 1.0 and 1.5 mg/L.
The 48h-EC50 was 0.60 mg/L with a 95% confidence interval between 0.52 and 0.76 mg/L. - Reported statistics and error estimates:
- The EC50 at the various observation times was determined using the maximum likelihood estimation method with the probits of the percentages of immobilised daphnids as function of the logarithms of the corresponding concentrations (Finney, D.J., 1971: Probit analysis, Cambridge University Press, Cambridge, U.K., 3rd edition)
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of the present study 2,4-Bismaleimidotoluene did not induce acute immobilisation of Daphnia magna at or below target concentrations of 1.0 mg/L, corresponding to a mean exposure concentration of 0.31 mg/L under flow-through test conditions (NOEC).
The 24h-EC50 for Daphnia was 0.71 mg/L with a 95% confidence interval between 0.60 and 0.94 mg/L (based on mean exposure concentrations).
The 48h-EC50 for Daphnia was 0.59 mg/L with a 95% confidence interval between 0.52 and 0.72 mg/L (based on mean exposure concentrations).
Reference
Table 1: Incidence of immobility in the additional range-finding test (flow-through):
Concentration Test substance; Target/Acutal (mg/L) |
Vessel Number |
Number Daphnia exposed |
Response at 24 h |
Response at 48 h |
||
number |
% |
number |
% |
|||
DMSO-control |
A |
5 |
0 |
0 |
0 |
0 |
B |
5 |
0 |
|
0 |
|
|
0.1/0.035 |
A |
5 |
0 |
0 |
0 |
0 |
B |
5 |
0 |
|
0 |
|
|
1.0/0.40 |
A |
5 |
2 |
20 |
3 |
70 |
B |
5 |
0 |
|
4 |
|
|
10/5.0 |
A |
5 |
4 |
70 |
5 |
100 |
B |
5 |
3 |
|
5 |
|
Table 2: Measured and average measured concentrations per test group (final test)
Concentration Target (mg/L) |
Measured T=0 (mg/L) |
Measured T=48 h (mg/L) |
Mean Measured (mg/L) |
0.10 |
0.054 |
0.064 |
0.059 |
0.22 |
0.074 |
0.096 |
0.085 |
0.46 |
0.188 |
0.193 |
0.19 |
1.0 |
0.277 |
0.343 |
0.31 |
2.2 |
0.755 |
0.883 |
0.82 |
Table 3:Acute immobilisation of Daphnia after 24 and 48 hours in the final EC50 study.
Concentration Test substance; Target/Mean (mg/L) |
Vessel Number |
Number Daphnia exposed |
Response at 24 h |
Response at 48 h |
||
number |
% |
number |
% |
|||
DMSO-control |
A |
5 |
0 |
0 |
0 |
0 |
B |
5 |
0 |
0 |
|||
C |
5 |
0 |
0 |
|||
D |
5 |
0 |
0 |
|||
0.10/0.059 |
A |
5 |
0 |
0 |
0 |
0 |
B |
5 |
0 |
0 |
|||
C |
5 |
0 |
0 |
|||
D |
5 |
0 |
0 |
|||
0.22/0.085 |
A |
5 |
0 |
0 |
0 |
0 |
B |
5 |
0 |
0 |
|||
C |
5 |
0 |
0 |
|||
D |
5 |
0 |
0 |
|||
0.46/0.19 |
A |
5 |
0 |
0 |
0 |
0 |
B |
5 |
0 |
0 |
|||
C |
5 |
0 |
0 |
|||
D |
5 |
0 |
0 |
|||
1.0/0.31 |
A |
5 |
0 |
0 |
0 |
0 |
B |
5 |
0 |
0 |
|||
C |
5 |
0 |
0 |
|||
D |
5 |
0 |
0 |
|||
2.2/0.82 |
A |
5 |
4 |
63 |
5 |
84 |
B |
4* |
3 |
3 |
|||
C |
5 |
4 |
5 |
|||
D |
5 |
1 |
3 |
*Inadvertently 4 instead of 5 Daphnids were exposed
Description of key information
- short-term toxicity to aquatic invertebrates: according to OECD TG 202 (adopted April 4th, 1984); RL 1; GLP; test with Daphnia magna 48 h EC50 = 591.4 µg/L
Key value for chemical safety assessment
Fresh water invertebrates
Fresh water invertebrates
- Effect concentration:
- 591.4 µg/L
Additional information
The 48-hr-acute toxicity of 2,4-Bismaleimidotoluene to Daphnia magna was studied under flow-through conditions according to OECD Guideline 202 (2004). Daphnids were exposed to control and test chemical at nominal concentrations of 0.10, 0.22, 0.46, 1.0 and 2.2 mg a.i./L (corresponding to 0.059, 0.085, 0.19, 0.31 and 0.82 mg/L mean measured concentrations) in tap water for 48 h. Mortality/immobilization was observed daily, pH was recorded at the beginning and at the end of the test and temperature was measured continuously. 2,4-Bismaleimidotoluene did not induce acute lmmobilisation of Daphnia magna at or below nominal concentrations of 1.0 mg/l, corresponding to a mean exposure concentration of 0.31 mg/l under flow-through test conditions (NOEC). The 48-hour EC50 was determined to be 0.59 mg a.i./L (95% confidence limits: 0.52 and 0.72 mg/L).
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