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EC number: 229-175-3 | CAS number: 6422-83-9
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- 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
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
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Toxicity to microorganisms
Administrative data
Link to relevant study record(s)
- Endpoint:
- activated sludge respiration inhibition testing
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2016-03-14 to 2016-12-08
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation))
- Version / remarks:
- 2010
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
A saturated solution was prepared: 180 mg/L were weighed out in ameasuring flask, filled up with demineralised water and stirred overnight. The saturated solution was filtered (45 µm). The concentration was determined by measurement of DOC (dissolved organic carbon) and was 43.7 mg/L. The DOC concentration was used to calculate the required volumes of saturated solution in the test vessels of the test item concentrations 0.1 mg/L (0.725 mL) and 0.56 mgIL (4.05 mL). The concentrations 3.2 mg/L, 18 mgIL and 100 mg/L was weighed out onto object slides and transferred with these into the Erlenmeyer flasks. 200 mL demineralised water was added and stirred overnight.
- Controls:
Test medium without test or reference item - Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- - Name and location of sewage treatment plant where inoculum was collected:
Municipal sewage treatment plant of 31137 Hildesheim, Germany
- Preparation of inoculum for exposure:
The sludge was washed twice with chlorine free tap water and adjusted to a dry sludge concentration of 3.0 g/L ± 10 %. The sludge was used within 24 h after sampling.
- Initial biomass concentration:
Dry sludge concentration: 3.05 g/L, corresponding to 1.53 g/L in the test vessel - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Hardness:
- n.a.
- Test temperature:
- 18 - 22°C
- pH:
- 7.5 ± 0.5
- Dissolved oxygen:
- > 60-70%
- Salinity:
- n.a.
- Conductivity:
- n.a.
- Nominal and measured concentrations:
- 0.1, 0.56, 3.2, 18 and 100 mg/L.
The concentration was determined by measurement of DOC (dissolved organic carbon). The DOC concentration was used to calculate the required volumes of saturated solution in the test vessels of the test item concentrations 0.1 mg/L and 0.56 mg/L - Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Material, size, headspace, fill volume: size: 1000 mL, fill volume: 500 mL
- Aeration: by shaking
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 6
- Sludge concentration (weight of dry solids per volume): 3.05 g/L, corresponding to 1.53 g/L in the test vessel
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: demineralised water
OTHER TEST CONDITIONS
- Adjustment of pH: not necessary
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Respiration Inhibition measured after 3h exposure
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 5.6
- Range finding study
- Test concentrations: 1st preliminary test: 10-100-1000 mg/L, 2nd preliminary test: 10 mg/L
- Results used to determine the conditions for the definitive study: Based on these results the definitive study was carried out with 5 concentrations within the range 0.1 - 100 mg/L in a geometrical series and with a dilution factor of 5.6 (above 100 mg/L, no increase in inhibition was determined in the 1st preliminary test). - Reference substance (positive control):
- yes
- Remarks:
- Copper (ll) sulphate pentahydrate p.a.
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- 1.93 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- inhibition of total respiration
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- 14.9 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- inhibition of total respiration
- Key result
- Duration:
- 3 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.56 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- inhibition of total respiration
- Results with reference substance (positive control):
- - Results with reference substance valid? yes
- Relevant effect levels: EC50 = 107 mg/L; 95% CI = 103-111 mg/L; recommended range of validity: 53-155 mg/L - Reported statistics and error estimates:
- One Way Analysis of Variance (ANOVA) and DUNNETT'S test was used for NOEC calculations. When running a One Way Analysis of Variance a Normality test and an Equal Variance test were done first. P-values for both Normality and Equal Variance test are 0.05. The EC-values of the test item and the reference item were calculated by sigmoidal dose-response regression and linear regression, respectively, using software GraphPadPrism.
- Validity criteria fulfilled:
- yes
- Conclusions:
- The present study was conducted according to OECD Test Guideline 209 in order to determine the respiration inhibition mediated by 2,4-Bismaleimidotoluene. Non-adapted activated sludge from a municipal sewage plant was incubated with 2,4-Bismaleimidotoluene for 3 h with the following concentrations: 0.1, 0.56, 3.2, 18 and 100 mg/L. The control was the test medium without test or reference item. Copper (ll) sulphate pentahydrate was used as reference substance and was incubated with 58, 100 and 180 mg/L. 2,4-Bismaleinimidotoluene is considered to be not toxic up to a concentration of 0.56 mg/L.
The following EC values and NOEC were determined:
EC10 = 1.93 mg/L; 95 % CI = 1.62-2.30
EC50 = 14.9 mg/L; 95 % CI = 13.1-17.1
NOEC = 0.56 mg/L: 95 % CI = -----
Reference
Validity of the study:
The study is valid as:
- the mean specific oxygen uptake rate of the control replicates was 21. 7 mg 02/g·h (validity
criterion: should be > 20 mg 02/g·h)
- the coefficient of variation of the oxygen uptake rates in the control replicates was 4.2 %
(validity criterion: < 30 %)
- the EC50 of the reference item was 107 mg/L and in the range of 53 - 155 mg/L
Table 1: Measured Oxygen uptake rate data
Treatment (mg a.i./L) [measured conc. used] |
Exposure time [3 h] |
||
Oxygen uptake rate [mg O2/L ×h] |
% Inhibition |
||
Control (dilution water only) |
Replicate #1 |
33.6 |
- |
Replicate #2 |
34.4 |
- |
|
Replicate #3 |
33.2 |
- |
|
Replicate #4 |
31.6 |
- |
|
Replicate #5 |
31.2 |
- |
|
Replicate #6 |
31.2 |
- |
|
0.1 |
Replicate #1 |
33.2 |
-2 |
Replicate #2 |
33.2 |
-2 |
|
Replicate #3 |
33.2 |
-2 |
|
Replicate #4 |
33.6 |
-3 |
|
Replicate #5 |
32.4 |
0 |
|
0.56 |
Replicate #1 |
32.4 |
0 |
Replicate #2 |
32.4 |
0 |
|
Replicate #3 |
30.8 |
5 |
|
Replicate #4 |
29.6 |
9 |
|
Replicate #5 |
31.2 |
4 |
|
3.2 |
Replicate #1 |
25.6 |
21 |
Replicate #2 |
26.0 |
20 |
|
Replicate #3 |
27.6 |
15 |
|
Replicate #4 |
28.0 |
14 |
|
Replicate #5 |
28.0 |
14 |
|
18 |
Replicate #1 |
14.8 |
54 |
Replicate #2 |
15.2 |
53 |
|
Replicate #3 |
14.8 |
54 |
|
Replicate #4 |
15.2 |
53 |
|
Replicate #5 |
14.8 |
54 |
|
100 |
Replicate #1 |
10.8 |
67 |
Replicate #2 |
10.4 |
68 |
|
Replicate #3 |
10.8 |
67 |
|
Replicate #4 |
10.4 |
68 |
|
Replicate #5 |
11.2 |
66 |
Description of key information
- Toxicity to microorganisms study; according to OECD 209; RL 1; GLP; EC50 = 14.9 mg/L, EC10 = 1.93 mg/L and NOEC = 0.56 mg/L.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 14.9 mg/L
- EC10 or NOEC for microorganisms:
- 1.93 mg/L
Additional information
The toxicity of 2,4-Bismaleimidotoluene to microorganisms was investigated in an activated sludge respiration inhibition test according to OECD guideline 209 (2010), respiration inhibition test using an inoculum obtained from non-adapted, activated sludge from a predominantly domestic sewage treatment plant over 48 h. The test was carried out under static conditions with the test item concentrations 0.1 -0.56 - 3.2 - 18 - 100 mg/L. The respiration rates of the control, reference and test item replicates were measured after a contact time of three hours, and the inhibitory effects of the test and reference item were determined in comparison to the control respiration rates. In order to check the activity of the test system and the test conditions a reference test was carried out with copper (ll) sulphate pentahydrate as reference item and the reference toxicity was determined.
The EC 50-value for the reference item was 107 mg/L. The mean inhibition of respiration for the test item replicates ranged from -2 % to 67 %. The EC10 was determined to be 1.93 mg/L, the EC50 was 14.9 mg/L and the NOEC was determined to be 0.56 mg/L.
Since, the coefficient of variation of the respiration rates of the control was < 30% and the oxygen uptakes rates of the control replicates were > 20 mg O2/g*h and the EC50-value of the reference was in the range of 53-155 mg/L the validity criteria were fulfilled. The test item had no inhibitory effect on oxygen uptake of sludge microorganisms up to a concentration of 0.56 mg/L, thus, the NOEC of 2,4-Bismaleimidotoluene is 0.56 mg/L. The determined EC50 is 14.9 mg/L (95 % CI = 13.1 -17.1) and the determined EC10 is 1.93 mg/L (95 % CI = 1.62 -2.30).
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