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EC number: 260-280-7 | CAS number: 56602-77-8
- 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:
- 23 February 2016 to 21 April 2016
- 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 (incl. QA statement)
- Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
The test material was sufficiently soluble in RO water to prepare an aqueous stock solution.
Therefore at test initiation 5 g (nominal) of the test material was weighed into a 2 L volumetric flask containing RO water. The pH of this solution was measured to be 11.87 and adjusted to 7.52 with concentrated HCL and 5N NaOH. - Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- - Preparation of inoculum for exposure:
A sample of activated sludge was obtained the day before the start of the test from Worlingworth Sewage Treatment Works (Suffolk, UK), which treats predominantly domestic waste. In the laboratory, the sample was maintained under aerobic conditions until required. The concentration of suspended solids in a homogenised sample was determined on the day of collection and immediately before the start of the test.
On the day of collection, an aliquot (10 mL) of the activated sludge was filtered through a dried and preweighed Whatman GF/C filter paper, which was then dried again at approximately 105 °C for at least one hour, allowed to cool in a desiccator and reweighed. The mixed liquor suspended solids (MLSS) content of the activated sludge was then calculated. Synthetic sewage (50 mL/L) was added to the stock of activated sludge and this was aerated overnight.
On the day of the test, the MLSS content of the sludge was determined (in triplicate) and adjusted to 4 g/L by the addition of tap water. The pH and temperature of the sludge were also measured. Aliquots (200 mL) were then added to each mixture to give a final MLSS concentration of 1.6 g/L in test vessels. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Test temperature:
- 20 ± 2 °C
- pH:
- Measurements of the pH of the aqueous stock solutions of the synthetic sewage, the test and reference substances and of the sample of activated sludge (4 g/L) before the start of the test are:
Synthetic sewage: 7.48
Test material stock solution: 7.52
Reference substance stock solution: 7.32
Activated sludge: 7.55 - Nominal and measured concentrations:
- Nominal concentrations: 10, 30, 100, 300 and 1 000 mg/L.
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 500 mL bottles fitted with an oxygen probe, a sinter to deliver air and a nitrogen gas nozzle.
- Material, size, headspace, fill volume: Mixtures were prepared to give a final volume of 500 mL.
- Aeration: The prepared mixtures were aerated and stirred for three hours in a thermostatically-controlled water bath, using an aerator connected to a laboratory supply of oil-free compressed air (one litre/minute).
- No. of vessels per concentration: Four replicates per concentration
- No. of vessels per control: Four replicates
- No. of vessels per reference control: One replicate per concentration
- Sludge concentration: 200 mL sludge inoculum was used per vessel.
- Nutrients provided for bacteria: Synthetic sewage feed for activated sludge was prepared by dissolving the following in 1 L of reverse-osmosis water:
16 g/L peptone
11 g/L meat extract
3 g/L urea
0.7 g/L sodium chloride
0.4 g/L calcium chloride dihydrate
0.2 g/L magnesium sulphate heptahydrate
2.8 g/L di-potassium hydrogen phosphate
The solution was stored in the fridge for up to 5 days before use. The pH of the solution was 7.5 ± 0.5.
16 mL of synthetic sewage was added to each vessel.
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water used to prepare solutions of test mixtures, synthetic sewage and the reference substance was reverse osmosis (RO) water.
OTHER TEST CONDITIONS
- Details on termination of incubation: Following the exposure period, the aeration was stopped and the headspace of each bottle was flushed with nitrogen during the oxygen measurement phase. The instrument measured the amount of oxygen in the mixtures at one minute intervals for at least 15 minutes.
EFFECT PARAMETERS MEASURED: Oxygen consumption
TEST CONCENTRATIONS
- Test concentrations: 10, 30, 100, 300 and 1 000 mg/L - Reference substance (positive control):
- yes
- Remarks:
- 3,5-dichlorophenol
- Key result
- Duration:
- 3 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Details on results:
- The mean specific respiration rate (Rs) of the control cultures incubated alongside the test mixtures was 22.9 mgO2/gh with a coefficient of variation (CV) of 23.2 %.
The mean respiration rates of samples of activated sludge exposed to the test material for three hours were significantly decreased at nominal concentrations of 300 and 1 000 mg/L, with respiration rates inhibited by 36 % and 52 %, respectively. The EC20 value was statistically determined to be 107 mg/L. Owing to the upper 95 % confidence limit being out of range, there was insufficient data for an EC50 value to be determined. However as there was 52 % inhibition the EC50 was estimated to be in the order of 1000 mg/L, the highest concentration tested. The no observed effect concentration (NOEC) for the test material was 100 mg/L. - Results with reference substance (positive control):
- The three-hour 50 % effect concentration (EC50) for 3,5-DCP was calculated to be 14.5 mg/L. These results show that the sample of activated sludge employed was sensitive to inhibition and that the test was valid.
- Reported statistics and error estimates:
- The statistical methods used in determining EC values are detailed in the statistical analysis report. The no observed effect concentration (NOEC) is the highest concentration tested that gave no statistically significant level of inhibition.
- Validity criteria fulfilled:
- yes
- Conclusions:
- The mean respiration rates of samples of activated sludge exposed to the test material for three hours were significantly decreased at nominal concentrations of 300 and 1000 mg/L, with respiration rates inhibited by 36 % and 52 %, respectively. The EC20 value was statistically determined to be 107 mg/L. Owing to the upper 95 % confidence limit being out of range, there was insufficient data for an EC50 value to be determined. However as there was 52 % inhibition the EC50 was estimated to be in the order of 1000 mg/L, the highest concentration tested. The no observed effect concentration (NOEC) for the test material was 100 mg/L.
- Executive summary:
The effect of the test material on the respiration rate of activated sludge was investigated in a study which was conducted under GLP conditions and in accordance with the standardised guideline OECD 209.
During the study samples of activated sludge (suspended solids 1.6 g/L), fed with synthetic sewage, were exposed to dilutions of the test material for three hours. Their rates of oxygen consumption were determined using oxygen electrodes and were compared with those of four replicate controls that contained activated sludge and synthetic sewage alone.
The nominal test material concentrations used in the study were 10, 30, 100, 300 and 1000 mg/L; four replicates were prepared for each concentration including the control. Single replicates of the reference inhibitor 3,5-dichlorophenol (3,5-DCP) were used at 3, 10 and 32 mg/L, as a positive control.
The mean specific respiration rate (Rs) of the control cultures incubated alongside the test mixtures was 22.9 mgO2/gh with a coefficient of variation (CV) of 23.2 %. The three-hour 50 % effect concentration (EC50) for 3,5-DCP was calculated to be 14.5 mg/L. These results show that the sample of activated sludge employed was sensitive to inhibition and that the test was valid.
The mean respiration rates of samples of activated sludge exposed to test material for three hours were significantly decreased at nominal concentrations of 300 and 1000 mg/L, with respiration rates inhibited by 36 % and 52 %, respectively. The EC20 value was statistically determined to be 107 mg/L. Owing to the upper 95 % confidence limit being out of range, there was insufficient data for an EC50 value to be determined. However as there was 52 % inhibition the EC50 was estimated to be in the order of 1000 mg/L, the highest concentration tested. The no observed effect concentration (NOEC) for the test material was 100 mg/L.
Reference
Description of key information
3-hour NOEC 100 mg/L, OECD 209, K Hillier
Key value for chemical safety assessment
- EC10 or NOEC for microorganisms:
- 100 mg/L
Additional information
The effect of the test material on the respiration rate of activated sludge was investigated in a study which was conducted under GLP conditions and in accordance with the standardised guideline OECD 209.
During the study samples of activated sludge (suspended solids 1.6 g/L), fed with synthetic sewage, were exposed to dilutions of the test material for three hours. Their rates of oxygen consumption were determined using oxygen electrodes and were compared with those of four replicate controls that contained activated sludge and synthetic sewage alone.
The nominal test material concentrations used in the study were 10, 30, 100, 300 and 1000 mg/L; four replicates were prepared for each concentration including the control. Single replicates of the reference inhibitor 3,5-dichlorophenol (3,5-DCP) were used at 3, 10 and 32 mg/L, as a positive control.
The mean specific respiration rate (Rs) of the control cultures incubated alongside the test mixtures was 22.9 mgO2/gh with a coefficient of variation (CV) of 23.2 %. The three-hour 50 % effect concentration (EC50) for 3,5-DCP was calculated to be 14.5 mg/L. These results show that the sample of activated sludge employed was sensitive to inhibition and that the test was valid.
The mean respiration rates of samples of activated sludge exposed to test material for three hours were significantly decreased at nominal concentrations of 300 and 1000 mg/L, with respiration rates inhibited by 36 % and 52 %, respectively. The EC20 value was statistically determined to be 107 mg/L. Owing to the upper 95 % confidence limit being out of range, there was insufficient data for an EC50 value to be determined. However as there was 52 % inhibition the EC50 was estimated to be in the order of 1000 mg/L, the highest concentration tested. The no observed effect concentration (NOEC) for the test material was 100 mg/L.
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