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EC number: 939-290-7 | CAS number: 68607-20-5
- 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
- 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:
- From 23 April, 2012 to 26 April, 2012
- 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
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- The test substance was added to the incubation vessels using a stock solution of approximately 1.0 g/L. The stock solution of of the test substance was prepared by dissolving 0.80 g in 800 mL deionized water.
pH stock solution of the test substance = 5.0 - Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- - Activated sludge
Unadapted secondary activated sludge was obtained (24-04-2012) from the WWTP Nieuwgraaf in Duiven. The WWTP Duiven is an activated sludge plant treating predominantly domestic wastewater. Prior to use the activated sludge was homogenized with a syringe. The dry weight of the homogenized activated sludge was determined and subsequently concentrated by settlement to the required dry weight concentration. The dry weight of the activated sludge in the incubation vessels was 1.5 g/L. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Test temperature:
- Temperature varied from 19.8 to 20.4°C.
- pH:
- The pH of the reaction mixtures after the incubation period ranged from 7.2 to 7.4.
- Dissolved oxygen:
- aerobic during test
- Details on test conditions:
- - Deionized water
Deionized water containing no more than 0.01 mg/L Cu and 1 mg/L non- purgeable organic carbon was prepared in a water purification system.
- Test conditions
The test was performed in 300 mL Erlenmeyer flasks with a total working volume of 50 mL. The homogenized activated sludge was incubated in a shaking water bath (100 rpm, 20°C) for 3 hours with various concentrations of the test compound and synthetic sewage.
- Synthetic sewage feed and stocks
The synthetic sewage feed was made by dissolving the following amounts of substances in 1 liter of deionized water: 16 g peptone, 11 g meat extract, 3 g urea, 0.7 g NaCl, 0.3 g CaCl2, 0.2 g MgSO4.7H2O, 2.8 g K2HPO4. In the incubation vessels 1.6 mL of the synthetic sewage feed is added in a total volume of 50 mL. The test substance and and 3,5-dichlorophenol were added to the incubation vessels using stock solutions both of approximately 1.0 g/L. The stock solution of the test substance was prepared by dissolving 0.80 g the test substance in 800 mL deionized water. The stock solution of 3,5-dichlorophenol was prepared by dissolving 0.1030 g 3,5-dichlorophenol in approximately 40 mL of deionized water with the addition of a few drops of 1 M NaOH. Finally the pH was corrected with 2M H2SO4 to approximately pH 7 and the solution was diluted further to 100 mL with deionized water.
-Test procedures
The activated sludge respiration inhibition test was performed according to the study plan. The study plan was developed from an OECD Test Guideline (OECD, 2010). In this test the inhibition of oxygen uptake by micro-organisms oxidizing organic carbon was not separately expressed from the respiration by micro- organisms oxidizing ammonium. The total respiration rate of activated sludge fed with a standard amount of synthetic sewage was measured. The respiration rate of the same activated sludge in the presence of various concentrations of the test substance under otherwise identical conditions was also measured. The inhibitory effect of the test substance at a particular concentration was expressed as a percentage of the mean respiration rates of the controls. Some aspects of the test are listed below:
• Prior to use the activated sludge was homogenized with a syringe.
• The activated sludge with test substances was incubated in a shaking bath (100 rpm/min) for 3 hours.
• The test substance and the reference compound (3,5-dichlorophenol) concentrations were spaced by a factor two.
• An abiotic control was not performed because neither reduction of oxygen by the test substance nor formation of oxygen due to chemical breakdown of the test substance is expected.
• No specific analyses were performed.
- Determination of respiration rates, pH, dry weight and temperature
The respiration rates of the activated sludge were measured in a Biological Oxygen Monitor (BOM) YSI 5300. The BOM consisted of a thermostated vessel with a magnetic stirrer and an oxygen electrode. The electrode to measure the oxygen depletion tightly closed the vessel. The volume of the vessel was 10 mL and the temperature in the vessel was 20°C. The pH was measured using an EUTECH pH meter. The temperature was measured with a Keithley 871A digital thermometer. The dry weight of the inoculum was determined by filtrating 50 mL of the activated sludge over a preweighed 12 m cellulose nitrate filter. This filter was dried for minimal 1.5 hours at 104 ± 5 °C and weighed after cooling. Dry weight was calculated by subtracting the weight of the filters and dividing the difference by the filtered volume.
- Calculation of test results
The respiration rate was calculated from the linear part of the respiration curve as mg O2/L/min. Divided by the activated sludge dry weight concentration in the incubation vessels a respiration rate (activity) in mg O2/g/min was calculated.
In order to calculate the inhibitory effect of a test substance at a particular concentration, the respiration rate was expressed as a percentage of the mean of two controls of the respiration rate:
(1 – 2Rs / (Rc1 + Rc2)) x 100 = percentage inhibition
Rs = oxygen-consumption rate at tested concentration of test substance.
Rc = oxygen-consumption rate of controls.
The EC values were computed from the best-fitted line (least square method) through the points given by the probit of the percentage inhibition and the logarithm of the concentration of the substance. All computations were performed with the program TOXCALCTM(Tidepool Scientific Software 1994- 1999). - Reference substance (positive control):
- yes
- Remarks:
- 3,5-dichlorophenol
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- 8 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC50
- Effect conc.:
- 35 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks:
- respiration rate
- Remarks on result:
- other: with 95% confidence limits of 2 and 163 mg/L
- Details on results:
- Test conditions
The pH of the reaction mixtures after the incubation period ranged from 7.2 to 7.4. Temperature varied from 19.8 to 20.4°C. These conditions allow
respiration of the activated sludge used.
Validity of the test
The validity of the test is demonstrated by three criteria. First, the coefficient of variation of the replicates of the control oxygen uptake rates was 10%, which fulfils the maximum prescribed variation of <30%. Secondly, the prescribed average oxygen uptake rate of > 20 mg O2/g dry weight/hour for the control measurements was achieved (30 mg O2/g dry weight/hour). The third criterion was met as shown by the EC50 of the reference compound of 15 mg/L, which is within the prescribed range of 2 to 25 mg/L.
Activated sludge respiration inhibition test
The inhibitory effect of the test substance at a particular concentration is expressed as a percentage of the two controls. From the results EC values were calculated. The EC50 of the test substance for activated sludge after 3 hours contact time is 35 mg/L with 95% confidence limits of 2 and 163 mg/L. The EC10, EC20 and EC80 are 8, 14 and 88 mg/L, respectively. The test substance therefore considered harmful to activated sludge.
Please refer to tables below. - Results with reference substance (positive control):
- The EC50 of the reference compound was 15 mg/L at a contact time of 3 hours, respectively. These EC50 values is within the prescribed range of 2 to 25 mg/L. Refer to the attachment for figure 1
- Validity criteria fulfilled:
- yes
- Conclusions:
- Under the study conditions, the 3 h EC50 of the test substance was determined to be 35 mg/L and the EC10, EC20 and EC80 were 8, 14 and 88 mg/L, respectively. The test substance is therefore considered harmful to activated sludge.
- Executive summary:
A study was conducted to determine the effect of the test substance, C16-18 ADBAC (98.2% active), on the respiration of activated sludge according to OECD 209 Guideline, in compliance with GLP. The toxicity to activated sludge was determined at a contact time of 3 h, using various concentrations (i.e., 2.5, 5, 10, 20 and 40 mg/L) of the test substance. No analytical determination was performed for the test substance. The total respiration rate of activated sludge fed with a standard amount of synthetic sewage was measured. The inhibitory effect of the test substance at a particular concentration was expressed as a percentage of the mean respiration rates of the controls. From the results EC values were calculated. Under the study conditions, the 3-h EC50 of the test substance for activated sludge was determined to be 35 mg/L with 95% confidence limits of 2 and 163 mg/L. The EC10, EC20 and EC80 were 8, 14 and 88 mg/L (nominal), respectively. The test substance is therefore considered harmful to activated sludge (Geerts, 2012).
Reference
TableI Respiration rates of theactivated sludge,inhibition percentages, pH and temperature at various concentrations of 3,5-dichlorophenol after 3 hours contact time.
Concentration (mg/L) |
Activity(mgO2/g/min) |
Inhibition (%) |
pH at start test |
pHatend test |
Temperature at end test (°C) |
Control |
0.5222 |
- |
7.0 |
7.2 |
19.8 |
Control |
0.5222 |
- |
7.1 |
7.2 |
20.0 |
Control* |
0.4556 |
- |
7.1 |
7.2 |
20.2 |
Control* |
0.4556 |
- |
7.1 |
7.3 |
20.4 |
2.5 |
0.5000 |
4 |
7.1 |
7.3 |
20.1 |
5.0* |
0.3778 |
17 |
7.1 |
7.2 |
20.0 |
10.0 |
0.3444 |
34 |
7.1 |
7.3 |
20.0 |
20.0 |
0.1500 |
71 |
7.0 |
7.3 |
20.2 |
40.0 |
0.1143 |
78 |
7.0 |
7.3 |
20.4 |
Concentrationstock solution3,5-dichlorophenol= 999mg/L
pHstock solution3,5-dichlorophenol=7.1
* = corresponding control and reference concentration
TableII Respiration rates of theactivated sludge, inhibition percentages, pHand temperature at various concentrations of the test substance after 3hours contact time.
Concentration (mg/L) |
Activity(mgO2/g/min) |
Inhibition (%) |
pH at start test |
pHatend test |
Temperature end test (°C) |
Control |
0.4556 |
- |
7.1 |
7.3 |
20.0 |
Control |
0.6133 |
- |
7.1 |
7.3 |
20.2 |
10 |
0.4810 |
10 |
7.1 |
7.3 |
20.2 |
20 |
0.3500 |
35 |
7.1 |
7.2 |
20.3 |
40 |
0.2296 |
57 |
7.1 |
7.3 |
20.1 |
80 |
0.1037 |
81 |
7.1 |
7.4 |
20.0 |
160 |
0.0556 |
90 |
7.1 |
7.4 |
20.1 |
Concentration stock solution the test substance = 1g/L;
pH stock solution of the test substance =5.0
Description of key information
The EC50 and EC10 values of the test substance for toxicity to microorganisms were determined to be 35 and 8 mg/L (nominal).
Key value for chemical safety assessment
- EC50 for microorganisms:
- 35 mg/L
- EC10 or NOEC for microorganisms:
- 8 mg/L
Additional information
A study was conducted to determine the effect of the test substance, C16-18 ADBAC (98.2% active), on the respiration of activated sludge according to OECD 209 Guideline, in compliance with GLP. The toxicity to activated sludge was determined at a contact time of 3 h, using various concentrations (i.e., 2.5, 5, 10, 20 and 40 mg/L) of the test substance. No analytical determination was performed for the test substance. The total respiration rate of activated sludge fed with a standard amount of synthetic sewage was measured. The inhibitory effect of the test substance at a particular concentration was expressed as a percentage of the mean respiration rates of the controls. From the results EC values were calculated. Under the study conditions, the 3-h EC50 of the test substance for activated sludge was determined to be 35 mg/L with 95% confidence limits of 2 and 163 mg/L. The EC10, EC20 and EC80 were 8, 14 and 88 mg/L (nominal), respectively. The test substance is therefore considered harmful to activated sludge (Geerts, 2012).
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