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EC number: 245-322-4 | CAS number: 22914-58-5
- 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
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- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
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- 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
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- Specific investigations
- Exposure related observations in humans
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- 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
- 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))
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- not required
- Details on sampling:
- Analytical monitoring not a requirement of the test guideline
- Vehicle:
- no
- Details on test solutions:
- The test item was dispersed directly in water.
Nominal amounts of test item (5.0, 16, 50, 160 and 500 mg (five replicates of each)) were each separately dispersed in approximately 200 mL of deionized reverse osmosis water and sealed with nescofilm prior to being subjected to ultrasonication for approximately 15 minutes followed by magnetic stirring for 24 hours, at room temperature, in order to maximize the dissolved test item concentration. All test vessels were shielded from the light during mixing.
Synthetic sewage (16 mL), activated sewage sludge (250 mL) and water were added to a final volume of 500 mL to give the required concentrations of 10, 32, 100, 320 and 1000 mg/L (five replicates of each). - Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- A mixed population of activated sewage sludge micro-organisms was obtained for the definitive test from the aeration stage of the Severn Trent Water Plc sewage treatment plant at Loughborough, Leicestershire, UK which treats predominantly domestic sewage.
The activated sewage sludge sample was maintained on continuous aeration in the laboratory at a temperature of approximately 21 ºC overnight prior to use in the test. On the day of collection the activated sewage sludge (14 liters) was fed synthetic sewage (700 mL). The pH of the sample on the day of the test was 7.3 measured using a Hach HQ40d Flexi handheld meter. Determination of the suspended solids level of the activated sewage sludge was carried out by filtering a sample (100 mL) of the activated sewage sludge by suction through a pre-weighed GF/A filter paper* using a Buchner funnel which was then rinsed three times with 10 mL of deionized reverse osmosis water and filtration continued for 3 minutes. The filter paper was then dried in an oven at approximately 105 ºC for at least 1-Hour and allowed to cool before weighing. This process was repeated until a constant weight was attained. The suspended solids concentration was equal to 3.0 g/L prior to use. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Remarks on exposure duration:
- Standard as per test guideline
- Hardness:
- Not specified
- Test temperature:
- 20 to 21°C
- pH:
- pH at the start and end of test in test vessels ranged from 7 to 7.9.
- Dissolved oxygen:
- The dissolved oxygen concentrations after 30 minutes contact time in all vessels were above 60% of the dissolved oxygen saturation level of 8.9 mg O2/L.
- Salinity:
- Not applicable
- Conductivity:
- Not specified
- Nominal and measured concentrations:
- Range-finder (nominal): 0, 10, 100 and 1000 mg/L
Definitive (nominal): 0, 10, 32, 100, 320 and 1000 mg/L - Details on test conditions:
- TEST SYSTEM
- Test vessel: 500 mL conical flask
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 500 mL (synthetic sewage, water and inoculum)
- Aeration: Yes
- No. of vessels per concentration (replicates): 5 replicates of each
- No. of vessels per control (replicates): 5
- No. of vessels per vehicle control (replicates): 6
- No. of vessels per abiotic control (replicates): NA
- Sludge concentration (weight of dry solids per volume): 3 g/L prior to use
- Nutrients provided for bacteria: Synthetic sewage meeting the OECD guideline requirements
- Nitrification inhibitor used (delete if not applicable): none
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The test water used for the range-finding and definitive tests was deionized reverse osmosis water containing less than 1 mg/L Total Organic Carbon (TOC).
- Particulate matter: None specified
OTHER TEST CONDITIONS
- Adjustment of pH: Yes adjusted as appropriate in a small number of replicates at varying concentrations at the start of study to get to the guideline specified pH values.
- Photoperiod: Continuous light
- Light intensity: Not specified
- Details on termination of incubation: As each vessel reached 3 hours contact time an aliquot was removed from the conical flask and poured into the measuring vessel (250 mL darkened glass Biological Oxygen Demand (BOD) bottle) and the rate of respiration measured using a Yellow Springs dissolved oxygen meter fitted with a BOD probe. The contents of the measuring vessel were stirred constantly by magnetic stirrer. The rate of respiration for each flask was measured over the linear portion of the oxygen consumption trace (where possible between 7 mg O2/L and 2 mg O2/L). In the case of a rapid oxygen consumption, measurements may have been outside this range but the oxygen consumption was always within the linear portion of the respiration curve. In the case of low oxygen consumption, the rate was determined over an approximate 10 minute period.
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : 3 hour calculation of percentage inhibition compared with controls.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: Standard as per OECD guidelines to enable determination of an appropriate EC50 and NOEC
- Justification for using fewer concentrations than requested by guideline: NA
- Range finding study
- Test concentrations: 0, 10, 100 and 1000 mg/L
- Results used to determine the conditions for the definitive study: Yes, results of the range-finder were used to determine the concentrations for the main phase of the study. - Reference substance (positive control):
- yes
- Remarks:
- 3,5-Dichlorophenol
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- 29 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- 360 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Duration:
- 3 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 32 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Details on results:
- No statistically significant toxic effects were shown at the test concentrations of 10 and 32 mg/L; however statistically significant toxic effects (P < 0.05) were shown at the test concentrations of 100, 320 and 1000 mg/L. Based on these results the NOEC after 3 hours exposure was 32 mg/L.
In some instances, the initial and final dissolved oxygen concentrations were outside those recommended in the test guidelines (7 mg O2/L and 2 mg O2/L respectively). This was considered to have had no adverse effect on the results of the study given that in all cases the oxygen consumption rate was determined over the linear portion of the oxygen consumption trace.
The dissolved oxygen concentrations after 30 minutes contact time in all vessels were above 60% of the dissolved oxygen saturation level of 8.9 mg O2/L.
The specific respiration rate of the controls was 29.78 mg oxygen per gram dry weight of sludge per hour and the coefficient of variation of oxygen uptake in the control vessels was 7.2%. - Results with reference substance (positive control):
- - Results with reference substance valid?
- Relevant effect levels: 3 hour EC50 7 mg/L. The EC50 (3-Hour contact time) for 3,5-dichlorophenol should lie within the range 2 to 25 mg/L for total respiration. - Reported statistics and error estimates:
- The percentage inhibition values were plotted against concentration, a line fitted using the Xlfit software package (IDBS) and the EC10, EC20, EC50 and EC80 values for the reference item and the EC10, EC20 and EC50 values for the test item determined from the equation for the fitted line. The EC80 value for the test item was determined by inspection of the inhibition of respiration rate data.
The 95% confidence limits were calculated for the reference item EC50 value using the method of Litchfield and Wilcoxon (Litchfield and Wilcoxon, 1949).
In order to determine the NOEC, one way analysis of variance incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981) and Dunnett's multiple comparison procedure for comparing several treatments with a control (Dunnett, 1955) was carried out on the oxygen consumption data after 3 hours to determine any statistically significant differences between the test and control groups. All statistical analyses were performed using the SAS computer software package (SAS, 1999 - 2001). - Validity criteria fulfilled:
- yes
- Conclusions:
- The effect of the test item on the respiration of activated sewage sludge micro-organisms gave a 3-Hour EC50 value of 360 mg/L. The NOEC after 3 hours exposure was 32 mg/L.
Reference
Inhibition of respiration rates:
|
Test substance |
3,5-DCP |
||
ECx mg/L (3 hours) |
95% confidence limits (mg/L) |
ECx mg/L (3 hours) |
95% confidence limits (mg/L) |
|
EC10 |
29 |
- |
1.4 |
- |
EC20 |
64 |
- |
2.1 |
- |
EC50 |
360 |
- |
7 |
5.3 – 9.2 |
EC80 |
> 1000 |
- |
2 |
- |
NOEC |
32 |
- |
- |
- |
Oxygen consumption rates and percentage inhibition values after 3 hours contact time in the definitive test
Nominal conc. (mg/L) |
Initial O2 reading (mgO2/L) |
Measurement period (minutes) |
Final O2 reading (mgO2/L) |
O2 Consumption rates (mgO2/L) |
Inhibition (%) |
|
Control |
R1 |
3.6 |
2 |
2 |
48 |
- |
R2 |
3.4 |
2 |
2 |
42 |
- |
|
R3 |
3.8 |
2 |
2.3 |
45 |
- |
|
R4 |
2.3 |
1 |
1.5 |
48 |
- |
|
R5 |
3.9 |
3 |
1.9 |
40 |
- |
|
R6 |
3.6 |
2 |
2.1 |
45 |
- |
|
Test substance |
10 R1 |
3.6 |
2 |
2.2 |
42 |
6 |
10 R2 |
4.0 |
2 |
2.5 |
45 |
[1] |
|
10 R3 |
4.3 |
3 |
2.1 |
44 |
1 |
|
10 R4 |
4.5 |
4 |
1.7 |
42 |
6 |
|
10 R5 |
4.6 |
2 |
3.1 |
45 |
[1] |
|
32 R1 |
4.1 |
4 |
1.5 |
39 |
13 |
|
32 R2 |
4.0 |
3 |
2 |
40 |
10 |
|
32 R3 |
3.8 |
3 |
1.8 |
40 |
10 |
|
32 R4 |
4.4 |
4 |
1.9 |
37.5 |
16 |
|
32 R5 |
3.3 |
2 |
1.9 |
42 |
6 |
|
100 R1 |
4.7 |
5 |
2 |
32.4 |
27 |
|
100 R2 |
5.4 |
7 |
2 |
29.14 |
35 |
|
100 R3 |
4.3 |
4 |
1.9 |
36 |
19 |
|
100 R4 |
4.6 |
6 |
1.6 |
30 |
33 |
|
100 R5 |
4.4 |
4 |
2 |
36 |
19 |
|
320 R1 |
5.8 |
9 |
1.8 |
26.67 |
40 |
|
320 R2 |
4.9 |
6 |
1.9 |
30 |
33 |
|
320 R3 |
6.1 |
10 |
2.4 |
22.2 |
50 |
|
320 R4 |
6.3 |
10 |
3.4 |
17.4 |
61 |
|
320 R5 |
5.9 |
9 |
3 |
19.33 |
57 |
|
1000 R1 |
6.5 |
10 |
3.6 |
17.4 |
61 |
|
1000 R2 |
6.6 |
10 |
4.1 |
15 |
66 |
|
1000 R3 |
6.4 |
10 |
4.1 |
13.8 |
69 |
|
1000 R4 |
5.8 |
10 |
2.9 |
17.4 |
61 |
|
1000 R5 |
6.7 |
10 |
4.5 |
13.2 |
70 |
|
3,5-DCP |
3.2 |
4.5 |
5 |
1.9 |
31.2 |
30 |
10 |
6.2 |
9 |
3.5 |
18 |
60 |
|
32 |
7.3 |
10 |
6.3 |
6 |
87 |
Description of key information
The effect of the test item on the respiration of activated sewage sludge micro-organisms gave a 3-Hour EC50 value of 360 mg/L. The NOEC after 3 hours exposure was 32 mg/L.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 360 mg/L
- EC10 or NOEC for microorganisms:
- 32 mg/L
Additional information
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