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EC number: 202-815-9 | CAS number: 100-06-1
- 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:
- 2017-05-09 to 2017-05-23
- 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:
- adopted 2010
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
- Version / remarks:
- 2015
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- no
- Details on sampling:
- - Sampling method for measurement of the respiration rate: well-mixed sample of each incubation mixture poured into a 250 mL glass BOD bottle after three hours incubation time (not further aerated)
- Sample storage conditions before analysis: the dissolved oxygen (DO) concentration was measured subsequently after sampling - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: At the start of the test, 16 mL synthetic sewage feed were made up to 250 mL with deionized water and then 250 mL activated sludge inoculum with a sludge concentration of nominally 3 g/L (dry weight) were added to this first control vessel. Thereafter, in time intervals of about fifteen minutes (an arbitrary but convenient interval) the procedure was repeated, except that the 16 mL synthetic sewage feed were added to the appropriate amounts of reference item or test item in deionized water to obtain a volume of 250 mL, followed by the addition of 250 mL inoculum. The test item was mixed into the deionized water by intense stirring for 15 minutes at room temperature to finely disperse and dissolve the maximum practical amount of the test item. No emulsifiers or solvents were used. Then, 16 mL of synthetic sewage feed and 250 mL of the activated sludge inoculum were added.
- Controls: Negative control, containing activated sludge and synthetic medium but no test or reference item; positive control, containing 3.2, 10 and 32 mg/L 3,5-dichlorophenol, activated sludge and synthetic medium
- Evidence of undissolved material: The test media for the final test concentrations up to and including 100 mg/L appeared as clear solutions. For the higher loading rates the test item was not completely dissolved under the test conditions. - Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- - Name and location of sewage treatment plant where inoculum was collected: ARA Birs (Birsfelden / Switzerland); aeration stage
- Preparation of inoculum for exposure: washed three times by centrifugation, decantation of the supernatant liquid phase and resuspension of the solid material in chlorine free tap water, dried thereafter - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Post exposure observation period:
- Not performed.
- Hardness:
- Not measured.
- Test temperature:
- 20 - 22 ºC.
- pH:
- - Negative control: 7.3 - 8.0
- Test item: 7.3 - 8.1
- Positive control: 7.3 - 8.6 - Dissolved oxygen:
- - Negative control: 7.3 - 8.5 mg/L
- Test item: 6.9 - 8.4 mg/L
- Positive control: 7.2 - 8.6 mg/L - Salinity:
- Not measured
- Conductivity:
- Not measured
- Nominal and measured concentrations:
- - Nominal: 10, 32, 100, 320 and 1000 mg/L
- Measured: not performed - Details on test conditions:
- TEST SYSTEM
- Test vessel: 2-L beakers
- Fill volume: 500 mL
- Aeration: yes, maintained by magnetic stirrers
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 1 for every concentration in positive control, 6 replicates for blank control
- Sludge concentration (weight of dry solids per volume): 3 g dry material per liter
- Nutrients provided for bacteria: peptone, meat extract, urea, NaCl, CaCl2, MgSO4 , K2HPO4
- Nitrification inhibitor used: N-allylthiourea
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water, containing less than 1 mg/L DOC (dissolved organic carbon) and chlorine free tap water (for preparation of inoculum)
OTHER TEST CONDITIONS
- Adjustment of pH: prior the test from pH 6.9 to pH 7.5 with a sodium hydroxide solution
- Light intensity: normal laboratory lighting
EFFECT PARAMETERS MEASURED:
TEST CONCENTRATIONS
- Range finding study: yes
- Test concentrations: 10, 32, 100, 320 and 1000 mg/L, - Reference substance (positive control):
- yes
- Remarks:
- 3,5-dichlorophenol
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- 403 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks on result:
- other: 95% CI: 270-660 mg/L
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- 43 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks on result:
- other: 95% CI: 11-83 mg/L
- Duration:
- 3 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Details on results:
- VALIDITY OF THE TEST
- The specific respiration rate of the blank controls was in the range of 37 to 46 mg oxygen per gram dry weight of sludge per hour. (Validity criterion: at least 20 mg O2/gh.)
- The coefficient of variation of oxygen uptake rate in control replicates was 7 %. (Validity criterion: less than 30 %).
- The 3-hour EC50 of the reference item 3,5-dichlorophenol was 9 mg/L. (Validity criterion:
2 to 25 mg/L).
The test is regarded as valid.
DOSE-RESPONSE RELATIONSHIP
A clear concentration effect relationship on the inhibition of the respiration rate was observed. - Results with reference substance (positive control):
- - Results with reference substance: valid
- Relevant effect levels: 3-hour EC50 = 9 mg/L (the 95 % confidence limits could not be calculated due to mathematical reasons) - Reported statistics and error estimates:
- The percentage inhibition of the respiration was plotted against logarithm of the test substance concentration using suitable software ToxRat® Professional. The NOEC was derived by per-vessel statistics using the Williams t-test (one sided smaller, α = 0.05).
The 3-hour ECx values and their 95 % confidence limits were calculated, as far as possible, by Probit Analysis. The 3-hour EC50 of the reference item 3,5-dichlorophenol was calculated by Probit Analysis, too.
All test results were based on nominal concentrations of the test and reference item. - Validity criteria fulfilled:
- yes
- Conclusions:
- The test item had a clear concentration-effect relationship on the inhibition of the respiration rate of activated sludge after the incubation period of three hours. The 3-hour EC50 value was 403 mg/L.
- Executive summary:
The inhibitory effect of the test item on the respiration rate of aerobic wastewater micro-organisms of activated sludge was investigated under GLP-conditions in a 3-hour respiration inhibition test according to the OECD Guidelines for Testing of Chemicals, No. 209, adopted 2010 and to the Commission Regulation (EU) No. 2016/266, C.11. (2015). Based on the results of range-finding test a definitive test with the following nominal test item concentrations was performed: 10, 32, 100, 320 and 1000 mg/L, with three replicates per concentration. As a positive control, the reference item 3,5-dichlorophenol was tested under identical test conditions, at concentrations of 3.2, 10 and 32 mg/L (all single replicates). Six blank controls, containing an equal volume of activated sludge and synthetic medium but no test or reference item, were included in the definitive test. The vessels were prepared in the following order: two blank controls, the reference item group (with ascending concentrations), two further blank controls, the test item group (with ascending concentrations) and finally again two blank control vessels. A clear concentration-effect relationship on the inhibition of the respiration rate was observed. The 3-hour NOEC was 10 mg/L and the EC50 value resulted in 403 mg/L (95% CI: 270 - 660).
The results of the blank and positive control treatments confirmed the suitability of the activated sludge and the method used. All the validity criteria were fulfilled.
Reference
Table 1: Influence of the test item and 3,5-Dichlorophenol (Reference Item) on the Oxygen Consumption of Activated Sludge.
Vessel no. |
Nominal concentration of test chemical |
Oxygen consumption rate (R) |
Specific respiration rate (Rs) |
Inhibition |
pH values |
Oxygenconcentration [mg O2/L] |
|||
[mg/L] |
[mg O2 /Lh] |
[mg O2 /gh] |
% |
start* |
end* |
start* |
end* |
||
Control |
|||||||||
1 |
0 |
66.93 |
44.62 |
|
|
7.6 |
8.0 |
8.5 |
7.5 |
2 |
0 |
63.87 |
42.58 |
|
|
7.4 |
7.9 |
8.5 |
7.3 |
6 |
0 |
68.67 |
45.78 |
|
|
7.3 |
7.8 |
8.3 |
7.3 |
7 |
0 |
68.40 |
45.60 |
|
|
7.3 |
8.0 |
8.4 |
7.3 |
23 |
0 |
56.07 |
37.38 |
|
|
7.3 |
7.9 |
8.2 |
7.4 |
24 |
0 |
68.25 |
45.50 |
|
|
7.3 |
7.8 |
8.2 |
7.3 |
Mean |
65.36 |
43.58 |
|
|
|
|
|
|
|
Standard Deveation |
4.89 |
3.26 |
|
|
|
|
|
|
|
Coefficient Variation |
7.48% |
7.48% |
|
|
|
|
|
|
|
Test item |
Mean |
||||||||
8 |
10 |
64.80 |
43.20 |
0.9 |
0.4 |
7.3 |
8.0 |
8.4 |
7.5 |
9 |
10 |
67.80 |
45.20 |
-3.7 |
7.4 |
7.7 |
8.3 |
6.9 |
|
10 |
10 |
62.64 |
41.76 |
4.2 |
7.3 |
8.0 |
8.4 |
7.6 |
|
11 |
32 |
52.91 |
35.27 |
19.1 |
13.4 |
7.3 |
8.0 |
8.4 |
7.5 |
12 |
32 |
61.20 |
40.80 |
6.4 |
7.3 |
7.9 |
8.4 |
7.4 |
|
13 |
32 |
55.64 |
37.09 |
14.9 |
7.4 |
7.9 |
8.3 |
7.4 |
|
14 |
100 |
54.00 |
36.00 |
17.4 |
18.9 |
7.5 |
7.9 |
8.3 |
6.9 |
15 |
100 |
52.50 |
35.00 |
19.7 |
7.4 |
8.0 |
8.3 |
7.5 |
|
16 |
100 |
52.47 |
34.98 |
19.7 |
7.3 |
8.0 |
8.2 |
7.0 |
|
17 |
320 |
37.13 |
24.76 |
43.2 |
42.0 |
7.3 |
8.0 |
8.4 |
8.1 |
18 |
320 |
36.57 |
24.38 |
44.1 |
7.3 |
8.0 |
8.2 |
7.9 |
|
19 |
320 |
40.05 |
26.70 |
38.7 |
7.4 |
8.0 |
8.2 |
7.6 |
|
20 |
1000 |
17.95 |
11.96 |
72.5 |
72.6 |
7.4 |
8.0 |
8.2 |
8.3 |
21 |
1000 |
17.51 |
11.67 |
73.2 |
7.4 |
8.1 |
8.2 |
8.4 |
|
22 |
1000 |
18.27 |
12.18 |
72.0 |
7.3 |
8.1 |
8.2 |
8.1 |
|
Reference item |
|||||||||
3 |
3.2 |
55.65 |
37.10 |
14.9 |
|
7.5 |
7.9 |
8.6 |
7.2 |
4 |
10 |
28.20 |
18.80 |
56.9 |
7.3 |
7.9 |
8.5 |
7.8 |
|
5 |
32 |
8.62 |
5.75 |
86.8 |
7.5 |
8.0 |
8.6 |
8.2 |
*: Start and end of the 3-hour incubation period.
- % inhibition: Negative value due to increased oxygen consumption rate relative to the control.
n.a.: Not applicable
Table 2: The 3-hour NOEC and ECX-values and their 95 % confidence limits
Endpoint after 3 Hours |
[mg/L] |
NOEC |
10 |
EC10 95 % confidence limits |
43 11 – 83 |
EC20 95 % confidence limits |
93 37 – 152 |
EC50 95 % confidence limits |
403 270 - 660 |
EC80 |
>1000 |
Description of key information
The toxicity of the test item to microorganisms was determined in a test according to OECD guideline 209 and Commission Regulation (EU) No. 2016/266, C.11.
The test determined an EC50 value of 403 mg/L and a NOEC of 10 mg/L.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 403 mg/L
- EC10 or NOEC for microorganisms:
- 43 mg/L
Additional information
The inhibitory effect of the test item on the respiration rate of aerobic wastewater microorganisms of activated sludge was investigated under GLP-conditions in a 3-hour respiration inhibition test according to the OECD Guidelines for Testing of Chemicals, No. 209, adopted 2010 and to the Commission Regulation (EU) No. 2016/266, C.11. (2015). Based on the results of range-finding test a definitive test with the following nominal test item concentrations was performed: 10, 32, 100, 320 and 1000 mg/L, with three replicates per concentration. As a positive control, the reference item 3,5-dichlorophenol was tested under identical test conditions, at concentrations of 3.2, 10 and 32 mg/L (all single replicates). Six blank controls, containing an equal volume of activated sludge and synthetic medium but no test or reference item, were included in the definitive test. The vessels were prepared in the following order: two blank controls, the reference item group (with ascending concentrations), two further blank controls, the test item group (with ascending concentrations) and finally again two blank control vessels. A clear concentration-effect relationship on the inhibition of the respiration rate was observed. The 3-hour NOEC was 10 mg/L and the EC50 value resulted in 403 mg/L (95% CI: 270 - 660). The results of the blank and positive control treatments confirmed the suitability of the activated sludge and the method used. All validity criteria were fulfilled.
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