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EC number: 260-124-8 | CAS number: 56358-09-9
- 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
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:
- 12-28.11.2012
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- PREPARATION OF TEST SOLUTION (especially for difficult test substances)
The test substance is non-soluble in water in concentrations needed for the test. For this reason direct dosing of the test substance was used. The substance was weighed on a slide and immersed into 100 mL of deionised water in the reaction container. - Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- - Name and location of sewage treatment plant where inoculum was collected: The activated biological sludge containing a mixed culture of microorganisms was obtained from the second step of the sewage treatment plant of Pardubice. The wastewater processed by the sewage treatment plant is predominantly municipal.
- Preparation of inoculum for exposure: The activated sludge was collected two days before the day of testing. After the sample collection the sludge was washed with potable water for 0.5 hours and subsequently decanted for 0.5 hours. This procedure was repeated three times in total. 50 mL of cultivation medium was added per 2 L of diluted sludge suspension at permanent aeration till the day of test.
- Pre-treatment: The pH was adjusted to 6.0 and the sludge suspension was aerated until use.
- Initial biomass concentration: The dry weight was determined from 10 mL of sludge suspension after 0.5 hours of sedimentation. Before the test the sludge was suspended in water up to a concentration of about 4000 mg of sludge dry weight per litre. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Test temperature:
- 20 ± 2 °C
- pH:
- approx. 6 – 8
- Nominal and measured concentrations:
- - Nominal concentrations: 100, 180, 320, 580 and 1000 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: Aerated apparatus with glass ends
- Type (delete if not applicable): Closed
- Material, size, headspace, fill volume: 500 mL; defined dose of the test or reference substance, 100 mL of deionised water, 16 mL of the synthetic nutrient medium and 200 mL of the prepared sludge suspension were introduced into each test container. The volume was replenished up to 500 mL with deionised water.
- Aeration: Yes, continuous aeration by filtered pressured air (passed through a filter for odour and oil removal)
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2
- No. of vessels per abiotic control (replicates): 1; performed at the highest concentration of the test substance and nutrient medium without sludge and with addition of 1 mL of 5 g/L mercury chloride solution.
- Sludge concentration (weight of dry solids per volume): Sludge concentration 18 030 mg/L
- Nutrients provided for bacteria: Synthetic sewage. Synthetic nutrient medium was always prepared fresh on the day of use. The following components were made up to 1 L with deionised water: peptone 16 g, meat extract 11 g, urea ((NH2)2CO) 3 g, sodium chloride (NaCl) 0.7 g, calcium chloride dehydrate (CaCl2.2H2O) 0.4 g, magnesium sulphate heptahydrate (MgSO4.7H2O) 0.2 g and anhydrous dipotassium hydrogen phosphate (K2HPO4) 2.8 g.
- Biomass loading rate: 4000 mg/L: 222 mL/L
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Deionised water
OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Daily
- Light intensity: Not specified
EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
The measurement of oxygen content was performed in aliquots of the reaction mixtures after the 3 hour incubation period. Bottles with narrow necks and a volume of about 280 mL (BOD bottles) were filled with the sample and positioned on a magnetic stirrer. Then, the oxygen electrode (equipped with a magnetic stirring wheel) was inserted in a way that no air remained in the bottle and that the electrode sealed the bottleneck to avoid contact of the sample with the atmosphere. The decline of oxygen concentration was measured and recorded at 10-minute intervals (respiration rate of activated sludge).
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study: Yes
- Test concentrations: 46, 100, 220, 480 and 1000 mg/L
- Results used to determine the conditions for the definitive study: Yes - Reference substance (positive control):
- yes
- Remarks:
- 3,5-dichlorophenol
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 1 000 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Details on results:
- In the main test, the test substance caused no inhibition of respiration. Abiotic decomposition was not detected even at the highest concentration of the test substance.
VALIDITY CRITERIA
- The respiration rates of two control experiments (at the beginning and at the end of the test) should not differ from each other more than 15 %.
The difference of the respiration rates of control experiments at the beginning and the end of the main test from each other was:
± 0.35 mg/L/h = ± 1.04 % (if average value of two controls 33.35 mg/L/h = 100 %)
± 0.69 mg/L/h = ± 2.10 % (if the value of lower control 33.00 mg/L/h =100 %)
- The EC50 value of the reference substance should be in the range of 5 to 30 mg/L. The EC50 value was 8.1 mg/L (95 % confidence interval: 6.9 to 9.2 mg/L)
The criteria of test validity were fulfilled. - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- Relevant effect levels: EC50 = 8.1 mg/L (95 % confidence interval 6.9 to 9.2 mg/L) - Reported statistics and error estimates:
- The values of EC50, EC20 and EC80 for the reference substance were calculated from the equation of regression line prepared by commercial software (Microsoft® Excel 2000).
The confidence intervals of EC50, EC20 and EC80 for the reference substance were calculated by the software QC.Expert 2.5 © 1998-2000 (product of TriloByte Ltd., Czech Republic).
The test substance caused no inhibition of respiration rate therefore it was not possible to calculate the EC50 value. - Validity criteria fulfilled:
- yes
- Conclusions:
- Under the conditions of this study, the EC50 was determined to be >1000 mg/L.
- Executive summary:
The influence of the test substance, Solvent Red 19E, on the respiration rate of activated sludge was investigated after a contact time of 3 hours under GLP conditions.
The test was performed according to Method C.11 - Activated Sludge Respiration Inhibition Test, Council Regulation (EC) No. 440/2008, published in O.J. L 142, 2008.
Following a preliminary test using 5 concentrations of the test substance in the range 46 to 1000 mg/L, in the main test, 5 concentrations of the test substance in geometric progression with factor of 1.8 were used to investigate the effects of the test substance. Concentrations were 100, 180, 320, 580 and 1000 mg/L. Activated sludge from a municipal sewage treatment plant was exposed to the test material along with two control experiments (one at the beginning and one at the end of the test) an abiotic control and 3,5-dichlorophenol as a reference substance.
In the main test, the test substance caused no inhibition of respiration. Abiotic decomposition was not detected even at the highest concentration of the test substance.
The test met all the validity criteria as the respiration rate of the controls did not differ too greatly and the EC50 of the reference substance was in the appropriate range.
Under the conditions of this study, the EC50 was determined to be >1000 mg/L. All tested concentrations in the main test caused no significant inhibition of respiration rate. Therefore it was not possible to calculate the EC50 value from available data.
Reference
Table 1: Calculation of Inhibition in the Main Test
Substance |
Concentration (mg/L) |
P1 (mg/L) |
P2 (mg/L) |
T1 (minutes) |
T2 (minutes) |
R (mg/L/h) |
Inhibition (%) |
Control |
0 |
6.46 |
2.61 |
3.0 |
10.0 |
33.00 |
0.0 |
3,5-DCP |
20.0 |
9.08 |
8.46 |
1.0 |
10.0 |
4.13 |
87.6 |
14.0 |
8.43 |
7.20 |
0.5 |
10.0 |
7.77 |
76.7 |
|
10.0 |
8.54 |
6.56 |
1.0 |
10.0 |
13.20 |
60.4 |
|
7.3 |
8.09 |
5.01 |
0.5 |
10.0 |
19.45 |
41.7 |
|
5.2 |
8.28 |
4.7 |
0.5 |
10.0 |
22.61 |
32.2 |
|
Test Substance |
100 |
6.42 |
2.65 |
3.0 |
10.0 |
32.31 |
3.1 |
180 |
6.37 |
2.77 |
2.5 |
9.0 |
33.23 |
0.3 |
|
320 |
6.25 |
2.54 |
2.0 |
8.5 |
34.43 |
-.3. |
|
580 |
6.40 |
2.50 |
0.5 |
7.0 |
36.00 |
-8.0 |
|
1000 |
6.32 |
2.67 |
2.0 |
8.5 |
33.69 |
-1.0 |
|
Control |
0 |
627 |
2.62 |
3.0 |
9.5 |
33.69 |
0.0 |
Abiotic Control |
1000 |
8.79 |
8.74 |
0.5 |
10.0 |
0.32 |
- |
3,5-DCP = 3,5-dichlorophenol
Average R value of the controls = 33.35 mg/L/h (SD = 0.49 mg/L/h)
Description of key information
The EC50 was determined to be >1000 mg/L.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 1 000 mg/L
Additional information
The influence of the test substance, Solvent Red 19E, on the respiration rate of activated sludge was investigated after a contact time of 3 hours under GLP conditions.
The test was performed according to Method C.11 - Activated Sludge Respiration Inhibition Test, Council Regulation (EC) No. 440/2008, published in O.J. L 142, 2008.
Following a preliminary test using 5 concentrations of the test substance in the range 46 to 1000 mg/L, in the main test, 5 concentrations of the test substance in geometric progression with factor of 1.8 were used to investigate the effects of the test substance. Concentrations were 100, 180, 320, 580 and 1000 mg/L. Activated sludge from a municipal sewage treatment plant was exposed to the test material along with two control experiments (one at the beginning and one at the end of the test) an abiotic control and 3,5-dichlorophenol as a reference substance.
In the main test, the test substance caused no inhibition of respiration. Abiotic decomposition was not detected even at the highest concentration of the test substance.
The test met all the validity criteria as the respiration rate of the controls did not differ too greatly and the EC50 of the reference substance was in the appropriate range.
Under the conditions of this study, the EC50 was determined to be >1000 mg/L. All tested concentrations in the main test caused no significant inhibition of respiration rate. Therefore it was not possible to calculate the EC50 value from available data.
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