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EC number: 257-913-4 | CAS number: 52434-90-9
- 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
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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:
- 12 April 2022 to 13 April 2022
- 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))
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- no
- Vehicle:
- no
- Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- The source of the inoculum of activated sludge was Sewage Treatment Plant, Eurofins Advinus Agrosciences Services India Private Limited, Bengaluru, India.
The inoculum was collected on 13 April 2022 at 08:45 AM from near the exit of the aeration tank of well operated wastewater treatment plant receiving predominantly domestic sewage. The sludge was allowed to settle for 15 minutes, and upper layer of finer solids was decanted. The collected sludge was centrifuged for 10 minutes at 3000 rpm and supernatant liquid was discarded. Then sludge was suspended in chlorine free tap water with shaking and re-centrifuge to remove wash-water and was discarded. Washing and centrifuging procedure was repeated three times.
A sample of washed sludge was weighed in triplicate and dried at 150°C in oven for 1 hour. From the obtained result the quantity of sludge required to obtain an activated sludge concentration of 3 g/L was calculated. During range finding test, 48.0601 g sludge was suspended in distilled water to make 6 L and 47.3400 g sludge was suspended in distilled water to make 6 L during definitive test. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 3 h
- Test temperature:
- 21.4 - 21.8 °C
- pH:
- Refer to tables 1 and 2 in section "Any other information on results incl. tables"
- Dissolved oxygen:
- Refer to tables 1 and 2 in section "Any other information on results incl. tables"
- Nominal and measured concentrations:
- Nominal: 1000 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 1000 mL beakers
- Aeration: forced aeration of 0.8-0.9 L/min to keep dissolved oxygen concentration above 60 to 70% saturation for 3 hours.
- No. of vessels per concentration (replicates): 3
- No. of vessels per blank control (replicates): 2
- No. of vessels per abiotic control (replicates): abiotic control was not included in the definitive test since there was no oxygen uptake at the highest test item concentration of 1000 mg/L during range finding test.
- Nutrients provided for bacteria: synthetic sewage feed as per OECD Guideline 209.
- Nitrification inhibitor used: N-allylthiourea, 3 replicates per concentration and 2 replicates per blank control.
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: distilled water containing <1 mg/mL TOC.
TEST CONCENTRATIONS
A range finding test was performed to determine the range of concentrations of test item to be used in the definitive test. The range finding test was performed with test item concentration at 10, 100 and 1000 mg/L along with a blank control and one abiotic control at 1000 mg/L test item concentration. For each group, one replicate was used.
Based on the results of the range finding test, the definitive test was carried out at the limit concentration of 1000 mg/L, with two sets of mixture to determine the total, heterotrophic and nitrification respiration inhibition. Since there was no oxygen uptake in the abiotic control at the highest test item concentration of 1000 mg/L during the range finding test, an abiotic control was not included in the definitive test. - Reference substance (positive control):
- yes
- Remarks:
- 3,5-Dichlorophenol
- Duration:
- 3 h
- Dose descriptor:
- NOEC
- Effect conc.:
- > 1 000 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of heterotrophic respiration
- 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 heterotrophic respiration
- Key result
- Duration:
- 3 h
- Dose descriptor:
- NOEC
- Effect conc.:
- > 1 000 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
- Details on results:
- The pH, oxygen consumption rate and percent inhibition of total respiration and heterotrophic respirations are presented in tables 1 and 2 under "Any other information on results incl. tables".
The pH of the mixtures after 3 hours incubation ranged from 8.19 to 8.27.
No significant inhibition of AP 729 was observed in both total respiration and heterotrophic respiration. The nitrification respiration rate was found to be not significant (significance was –0.268%). Hence the inhibition of AP 729 due to nitrification respiration was not calculated. - Results with reference substance (positive control):
- The reference substance 3,5-Dichlorophenol was tested at concentrations of 2, 8 and 32 mg/L.
The calculated percent inhibition for total respiration was 39.2, 61.7 and 95.1 % and the heterotrophic respiration was 23.5, 61.8 and 93.5 % at the tested concentrations, respectively.
The EC50 value of 3,5-Dichlorophenol for total respiration inhibition is 3.68 mg/L with fiducial limits at 95% ranging from 2.041 to 17.588 mg/L.
The EC50 value of 3,5-Dichlorophenol for heterotrophic respiration inhibition is 5.29 mg/L with fiducial limits at 95% ranging from 4.263 to 6.559 mg/L. - Reported statistics and error estimates:
- The percentage inhibition of total respiration, heterotrophic respiration and nitrifying respiration was analyzed with the statistical method of Probit analysis.
The dose response curve was plotted and the slope of the curves and the median effective concentration (EC50) with 95% confidence limits as per maximum likelihood method of Finney (1978) was calculated using a validated computer program. - Validity criteria fulfilled:
- yes
- Conclusions:
- The EC50 and NOEC values for total respiration inhibition and heterotrophic respiration inhibition of AP 729 are >1000 mg/L.
- Executive summary:
The toxic effect of AP 729 on microorganisms (largely bacteria) from activated sludge of a well operated wastewater treatment plant receiving predominantly domestic sewage was studied according to OECD Guideline 209.
A range finding test was performed to determine the range of concentrations of test item to be used in the definitive test. Based on the results of the range finding test, the tested concentration of AP 729 in the definitive test was at the limit concentration of 1000 mg/L in triplicate and in two sets, along with blank control (without test item) and reference substance (3,5-Dichlorophenol) at concentrations of 2, 8 and 32 mg/L. One set of all test groups was used to determine total respiration and another set of all test groups added with N-allylthiourea at 11.6 mg/L was used to determine the heterotrophic respiration and the nitrification potential of the sludge.
The test item mixtures and the reference substance mixtures were prepared by adding the required quantity of test item and reference substance directly into the test vessels to obtain different nominal concentrations, respectively.
Blank control, test mixtures and reference substance mixtures were aerated for the exposure time of 3 hours at a temperature between 21.4 and 21.8°C. The oxygen consumption rate in the mixture was measured for 10 minutes using dissolved oxygen electrode and the respiration rate was calculated.
No significant inhibition of AP 729 was observed in both total respiration and heterotrophic respiration.
Moreover, the nitrification respiration rate was found to be not significant: hence the inhibition of AP 729 due to nitrification respiration was not calculated.The validity criteria of the test were fulfilled as:
- The blank controls oxygen uptake rate was 20.146 mg oxygen per one gram of activated sludge (dry weight of suspended solids) in an hour, which is more than the required 20 mg/g. The coefficient of variation of oxygen uptake rate in control replicates was 0.52 % which is within allowed 30% at the end of test.
- The EC50 of 3,5-Dichlorophenol for total and heterotrophic respiration was 3.68 and 5.29 mg/L, which is in the range 2 to 25 mg/L for total respiration and 5 to 40 mg/L for heterotrophic respiration, respectively.
Therefore, the EC50 and NOEC (No Observed Effect Concentration) value for total respiration inhibition and heterotrophic respiration inhibition of AP 729 are higher than 1000 mg/L.
Reference
The pH, oxygen consumption rate and percent inhibition of total respiration and heterotrophic respirations of the definitive test are presented in the tables 1 and 2, respectively.
Group No. | Description (mg/L) | pH of mixture at the end of incubation | Oxygen consumption rate (mg/L min) | Oxygen consumption rate (mg/L h) | Specific respiration rate a (mg/g h) | Percent inhibition, total respiration |
G1 | Blank control at start of test | 8.20 | 0.5055 | 30.330 | 20.220 | - |
Test Item | ||||||
G2 | 1000 | 8.27 | 0.5073 | 30.438 | 20.292 | -0.7 |
G3 | 1000 | 8.26 | 0.5091 | 30.546 | 20.364 | -1.1 |
G4 | 1000 | 8.25 | 0.5082 | 30.492 | 20.328 | -0.9 |
Reference Item | ||||||
G5 | 2 | 8.24 | 0.3064 | 18.384 | 12.256 | 39.2 |
G6 | 8 | 8.21 | 0.1927 | 11.562 | 7.708 | 61.7 |
G7 | 32 | 8.23 | 0.0245 | 1.470 | 0.980 | 95.1 |
G8 | Blank control at end of test | 8.25 | 0.5018 | 30.108 | 20.072 | - |
Group No. | Description (mg/L) | pH of mixture at the end of incubation | Oxygen consumption rate (mg/L min) | Oxygen consumption rate (mg/L h) | Specific respiration rate a (mg/g h) | Percent inhibition, heterotrophic respiration |
G1N | Blank control at start of test | 8.19 | 0.5045 | 30.270 | 20.18 | - |
Test Item | ||||||
G2N | 1000 | 8.20 | 0.5082 | 30.492 | 20.328 | -0.6 |
G3N | 1000 | 8.23 | 0.5055 | 30.330 | 20.220 | -0.1 |
G4N | 1000 | 8.25 | 0.5082 | 30.492 | 20.328 | -0.6 |
Reference Item | ||||||
G5N | 2 | 8.27 | 0.3864 | 23.184 | 15.456 | 23.5 |
G6N | 8 | 8.22 | 0.1927 | 11.562 | 7.708 | 61.8 |
G7N | 32 | 8.24 | 0.0327 | 1.962 | 1.308 | 93.5 |
G8N | Blank control at end of test | 8.26 | 0.5055 | 30.330 | 20.220 | - |
Note: a: oxygen consumed per g dry weight of sludge per hour.
Description of key information
The toxic effects of AP 729 on microorganisms from activated sludge of a well operated wastewater treatment plant receiving predominantly domestic sewage were studied according to OECD Guideline 209.
The tested concentration of AP 729 was at the limit concentration of 1000 mg/L in triplicate and in two sets, along with blank control and reference substance (3,5-Dichlorophenol). One set of all test groups was used to determine total respiration and another set of all test groups added with N-allylthiourea at 11.6 mg/L was used to determine the heterotrophic respiration and the nitrification potential of the sludge.
Blank control, test mixtures and reference substance mixtures were aerated for the exposure time of 3 hours. No significant inhibition of AP 729 was observed in both total respiration and heterotrophic respiration.
Therefore, the EC50 and NOEC value for total respiration inhibition and heterotrophic respiration inhibition of AP 729 are >1000 mg/L.
Key value for chemical safety assessment
- EC50 for microorganisms:
- 1 000 mg/L
- EC10 or NOEC for microorganisms:
- 1 000 mg/L
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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