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EC number: 608-554-0 | CAS number: 3090-56-0
- 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:
- 2016-05-17 to 2016-05-31
- 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
- Version / remarks:
- 2010
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
- Version / remarks:
- 2008
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 230-2015
- Expiration date of the lot/batch: 17 Nov 2017 - Analytical monitoring:
- no
- Details on sampling:
- No analytical analysis was done.
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Test substance preparations: The test substance was added in the required amounts according to the test concentrations directly to the test vessels with 234 mL deionized water. 16 mL synthetic medium were added.
- Reference substance preparations: 1000.0 mg of the reference substance was added to about 800 mL of deionized water and stirred at room temperature until the reference substance was completely dissolved. The pH value of the stock solution was measured and adjusted to 7.1 with 1 M sodium hydroxide solution. Following this the stock solution was made up to 1L with deionized water. The stock solution appeared yellowish and no undissolved reference substance was visible. Aliquots of the reference substance stock solution were dosed to the test vessels and made up with deionized water to a volume of 234 mL. 16 mL synthetic medium were added.
- Control preparations: To prepare the blank control assays 234 mL of deionized water and 16 mL synthetic medium were mixed. - Test organisms (species):
- activated sludge, domestic
- Details on inoculum:
- - Name and location of sewage treatment plant where inoculum was collected: aeration tank of the wastewater treatment plant of Mannheim, Germany
- Method of cultivation: No cultivation on site. The sludge was used on the day after collection.
- Preparation of inoculum for exposure: After arrival of the activated sludge suspension in the test facility the suspension was sieved with a fine woven mesh (mesh size about 1 mm). This suspension was pre-aerated over night at room temperature. At the next day the sludge suspension was washed once with drinking water and the suspension was adjusted to 3 g/L dw.
- Pretreatment: No pretreatment was done.
- Initial biomass concentration: 3 g/L dw - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Test temperature:
- 19.0 – 19.9 °C
- pH:
- 7.5
- Dissolved oxygen:
- start of measurement: > 7 mg/L
- Salinity:
- Not applicable
- Nominal and measured concentrations:
- 1000, 500, 250, 125, 62.5 mg/L as nominal concentration without correction of purity and blank controls
100, 10, 1 mg/L as nominal concentration based on reference substance - Details on test conditions:
- TEST SYSTEM
- Test vessel: Glas-beakers (nominal volume 1L)
- Fill volume: 500 mL
- Aeration: Oxygen concentration during aeration: > 2 mg/L; Oxygen concentration immediately before measurement: > 7 mg/L; Duration of the measurement of oxygen consumption: About 8 - 10 min
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- No. of vessels per reference control (replicates): 2
- Sludge concentration: 1.5 g/L dw
- Nutrients provided for bacteria: None
- Nitrification inhibitor used: None
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water
OTHER TEST CONDITIONS
- Adjustment of pH: An adjustment was not necessary.
EFFECT PARAMETERS MEASURED:
The consumption rate (mg O2/L*x minutes) of each test concentration and the blank controls were determined after 3 hours of exposure. The consumption rates (mg O2/L*h) were used to calculate the inhibition effects at particular test concentrations compared to the mean consumption rate of all controls.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: ≤ 3.2
- Range finding study: Not conducted.
- Test concentrations selection: According to the test guidelines, at least 5 concentrations in a geometric series with a separation factor of ≤ 3.2 should be used, preferably encompassing the range from 10 – 80 % inhibition. - Reference substance (positive control):
- yes
- Remarks:
- 3,5 dichlorophenol
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- < 62.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- 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
- Duration:
- 3 h
- Dose descriptor:
- other: EC20
- Effect conc.:
- 210 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Duration:
- 3 h
- Dose descriptor:
- other: EC80
- Effect conc.:
- > 1 000 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Details on results:
- - Any observations that might cause a difference between measured and nominal values: No observations reported.
- Effect concentrations exceeding solubility of substance in test medium: No.
- Adsorption: No.
- Blank controls oxygen uptake rate: 25 mg/g*h
- Coefficient of variation of oxygen uptake rate in control replicates: 2.2 % O2 consumption
- Outliers: The inhibition in the test assays TS4 – TS6 (test concentration 125 mg/L) was about 50 % and higher than the test concentration 250, 500 and 1000 mg/L. This is not possible. Therefore the test assays TS4 – TS6 were set as outliers. The statistical evaluation were done without the oxygen consumption values of the the test assays TS4 – TS6. - Results with reference substance (positive control):
- - Results with reference substance valid: yes
- Relevant effect levels: The EC50 of 3,5-dichlorophenol was 7.0 mg/L (OECD 209 range: 2-25 mg/L after 3 hours of incubation) - Reported statistics and error estimates:
- The consumption rates were used for the determination of the ECx by the probit method based on Finney with the software TOXRAT Professional 2.10. The effect concentrations were given with an accuracy of 2 significant digits.
- Validity criteria fulfilled:
- yes
Reference
Measured data of oxygen content of blank control and reference substance assays at the start and end of the evaluation time
Assay idenification |
Blank Control |
Reference Substance |
||||||||||
Replicate |
1 |
2 |
3 |
4 |
5 |
6 |
1 |
2 |
3 |
4 |
5 |
6 |
O2 conc. Start [mg/L] |
8.4 |
7.5 |
7.5 |
8.0 |
8.0 |
8.0 |
7.6 |
8.0 |
9.0 |
8.9 |
8.9 |
9.1 |
O2 conc. End [mg/L] |
4.5 |
3.9 |
3.9 |
4.4 |
4.4 |
4.5 |
4.0 |
4.5 |
7.7 |
7.5 |
8.6 |
8.8 |
O2 consumption rate [mg/L] |
3.9 |
3.6 |
3.6 |
3.6 |
3.6 |
3.5 |
3.6 |
3.5 |
1.3 |
1.4 |
0.3 |
0.3 |
O2 consumption in minutes |
6.2 |
6.0 |
5.9 |
5.9 |
6.0 |
5.9 |
5.9 |
6.0 |
6.0 |
6.0 |
6.0 |
5.9 |
O2 consumption rate [mg/L×h], RT |
38 |
36 |
37 |
37 |
36 |
36 |
37 |
35 |
13 |
14 |
3 |
3 |
Specific O2 consumption rate [mg/g×h] |
25 |
24 |
25 |
25 |
24 |
24 |
25 |
23 |
9 |
9 |
2 |
2 |
Calculation of inhibition respiration [%], IT |
- |
-1 |
5 |
65 |
62 |
92 |
92 |
The respiration rate was calculated in mg O2/L×h. The oxygen consumption was
measured over a period of 8-10 minutes. 5.9 - 6.2 minutes from the plotted track within a linear range of the graph was used for the determination of the respiration rate.
Measured data of oxygen content of the test assays at the begin and end of the
evaluation time
Assay idenification |
Test Assay |
||||||||||||||
Replicate |
1 |
2 |
3 |
4* |
5* |
6* |
7 |
8 |
9 |
10 |
11 |
12 |
13 |
14 |
15 |
O2 conc. Start [mg/L] |
8.0 |
7.8 |
7.5 |
8.5 |
8.5 |
8.2 |
7.5 |
7.5 |
7.7 |
7.9 |
7.5 |
8.0 |
7.7 |
7.5 |
7.3 |
O2 conc. End [mg/L] |
5.0 |
4.5 |
4.4 |
6.5 |
6.6 |
6.3 |
4.5 |
4.6 |
4.5 |
5.3 |
4.8 |
5.4 |
5.2 |
4.9 |
4.7 |
O2 consumption rate [mg/L] |
6.2 |
6.0 |
5.9 |
6.5 |
6.1 |
6.1 |
6.0 |
6.0 |
6.1 |
6.1 |
6.1 |
6.0 |
6.0 |
6.0 |
6.0 |
O2 consumption in minutes |
3.0 |
3.3 |
3.1 |
2.0 |
1.9 |
1.9 |
3.0 |
2.9 |
3.2 |
2.6 |
2.7 |
2.6 |
2.5 |
2.6 |
2.6 |
O2 consumption rate [mg/L×h], RT |
29 |
33 |
32 |
18 |
19 |
19 |
30 |
29 |
31 |
26 |
27 |
26 |
25 |
26 |
26 |
Specific O2 consumption rate [mg/g×h] |
19 |
22 |
21 |
12 |
13 |
13 |
20 |
19 |
21 |
17 |
18 |
17 |
17 |
17 |
17 |
Calculation of inhibition respiration [%], IT |
21 |
10 |
13 |
51 |
48 |
48 |
18 |
21 |
15 |
29 |
26 |
29 |
32 |
29 |
29 |
The respiration rate was calculated in mg O2/L×h. The oxygen consumption was
measured over a period of 8-10 minutes. 6 - 6.1 minutes from the plotted track within a linear range of the graph was used for the determination of the respiration rate.
Equation1: IT=(1-(RT/RTB)*100
For calculating the inhibition the results of the fractions in eq.1 were rounded to
two decimal places.
IT= inhibition of the total respiration [%]
RT = oxygen consumption in the test assay [mg O2/L×h]
RTB= mean value oxygen consumption of all blank controls [mg O2/L×h]
Description of key information
The toxic effects of the test substance on the oxygen consumption rate of aerobic microorganisms (activated sludge) after short-term exposure of 3 h were determined according to the OECD 209 (2010). The EC50 was determined to be > 1000 mg/L (nominal), the EC20 was determinded to be 210mg/L (nominal)
and the EC10 was determined to be < 62.5 mg/L(nominal).Key value for chemical safety assessment
- EC50 for microorganisms:
- 1 000 mg/L
Additional information
The study according to OECD 209 (2010) was conducted to determine the toxic effects of the test substance on the oxygen consumption rate of aerobic microorganisms (activated sludge). The exposure duration was 3 h and the test concentrations were 1000, 500, 250, 125, 62.5 mg/L (nominal, without correction of the purity). No measurements of the test concentrations were done. The activated sludge was derived from a municipal waste treatment plant. The sludge concentration was 1.5 g/L and a synthetic medium according to the guideline was used. A reference substance (3,5-dichlorphenol) was used as positive control. The EC50 value of the reference substance 3,5-dichlorphenol was determined to be 7.0 mg/L, which was within the acceptable limits (2 to 25 mg/L), as specified in OECD 209 Guideline. In addition the coefficient of variation of the six replicates of blank control was determined to be 2.2 % O2 consumption. The mean oxygen uptake of the blank controls was determined to be 25 mg/g*h. Based on these results it can be stated that the test was valid. For the test substance the EC50 was determined to be > 1000 mg/L (nominal), the EC20 was determinded to be 210mg/L (nominal) and the EC10 was determined to be < 62.5 mg/L (nominal).
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