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EC number: - | CAS number: -
- 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:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- August 2019 - October 2019
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Reason / purpose for cross-reference:
- read-across source
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
- Version / remarks:
- OECD (2010)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- Chemical name: Substituted amino acid (2) solution FC-C 13587
Assay: 27.8% active; 72.6% water
Batch/lot No.: FC-C 13587
Appearance: Clear colorless solution
Solubility: Soluble in water at test concentrations
Volatile: Expected non-volatile
Expiry date
March 2020 (if stored at max -18°C) - Analytical monitoring:
- no
- Vehicle:
- no
- Test organisms (species):
- activated sludge of a predominantly domestic sewage
- Details on inoculum:
- Activated sludge
Unadapted secondary activated sludge was obtained (28-08-2019) 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 measured dry weight of the homogenized activated sludge was 3.4 g/L and was concentrated by settlement to 6.0 g/L dry weight. Subsequently, the concentrated activated sludge was diluted in the incubation vessels to an activated sludge dry weight concentration of 1.5 g/L. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 3 h
- Remarks on exposure duration:
- max value according to Guideline
- Test temperature:
- The temperature of the BOM thermostated vessel measured at the start, middle and end of the test varied from 19.1 to 19.6 °C. The temperature of the water bath, used for the incubation of the activated sludge, at the beginning, middle and end of the test was 20.2 °C, 20.2 °C and 20.3 °C, respectively. Temperatures measured in the Erlenmeyer flasks at the end of the incubation period varied from 20.0 to 20.4 °C.
- pH:
- The pH of the reaction mixtures after the incubation period ranged from 7.0 to 7.3
- Dissolved oxygen:
- is measured
- Salinity:
- Freshwater
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, 0.4 g CaCl2.2H2O, 0.2 g MgSO4.7H2O, 2.8 g K2HPO4. In the incubation vessels 1.6 mL and 6.4 mL of the synthetic sewage feed is added in a total volume of 50 and 200 mL, respectively. - Details on test conditions:
- 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. - Reference substance (positive control):
- yes
- Remarks:
- 3,5-Dichlorophenol (p.a. quality 99.1%) used as a reference substance in the activated sludge respiration inhibition test was purchased from Sigma Aldrich.
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- 591 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- inhibition of total respiration
- Remarks on result:
- other: 95% CL: 276 - 2933 mg/L
- Key result
- Duration:
- 3 h
- Dose descriptor:
- EC10
- Effect conc.:
- 164.3 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- act. ingr.
- Basis for effect:
- inhibition of total respiration
- Remarks on result:
- other: 95% CL: 76.7 - 815.4 mg/L
- Details on results:
- Test conditions
The pH of the reaction mixtures after the incubation period ranged from 7.0 to 7.3. The temperature of the BOM thermostated vessel measured at the start, middle and end of the test varied from 19.1 to 19.6 °C. The temperature of the water bath, used for the incubation of the activated sludge, at the beginning, middle and end of the test was 20.2 °C, 20.2 °C and 20.3 °C, respectively. Temperatures measured in the Erlenmeyer flasks at the end of the incubation period varied from 20.0 to 20.4 °C. These conditions allow respiration of the activated sludge used.
Validity of the test
The validity of the test is demonstrated although only two out of three validity criteria were met according the guideline. First, the prescribed oxygen uptake rate of >20 mg O2/g dry weight/hour for the control measurements was not met. The inoculum used was obtained from a treatment plant which is operated at a low organic sludge load and as a consequence has a low oxygen uptake rate. Operating at low organic sludge loads is nowadays very common. The average oxygen uptake rate for the control replicates was 12 mg O2/g dry weight/hour, which is lower than the prescribed rate. This does not influence the sensitivity of the activated sludge as shown by comparable EC50 values of the reference substance 3,5-dichlorophenol at oxygen uptake rates lower and higher than the prescribed rate (AkzoNobel 2012). The lower average oxygen uptake rate does therefore not compromise the test outcome in terms of robustness of the endpoints obtained for the test and reference substance and does not invalidate the test. Second, the coefficient of variation of the replicates of the control oxygen uptake rates was 17%, which shows a high precision and fulfills the maximum prescribed variation of <30%.
Third, the EC50 of the reference compound for activated sludge after 3 hours contact time is 8 mg/L, which is within the prescribed range of 2 to 25 mg/L (Table I and Figure 1).
Activated sludge respiration inhibition test
The calculated respiration rates and inhibition percentages of substituted amino acid (2) solution FC-C 13587 are presented in Table II and the results of the graphical probit analysis are shown in Figure 2. The inhibitory effect of substituted amino acid (2) solution FC-C 13587 at a particular concentration is expressed as a percentage of the two controls. From the results EC values were calculated. The EC10 of substituted amino acid (2) solution FC-C 13587 for activated sludge after 3 hours contact time is 591 mg/L with 95% confidence limits of 276 and 2933 mg/L. The EC20, EC50 and EC80 values could not be determined. Substituted amino acid (2) solution FC-C 13587 is therefore considered not harmful to activated sludge. - Results with reference substance (positive control):
- The EC50 of the reference compound for activated sludge after 3 hours contact time is 8 mg/L, which is within the prescribed range of 2 to 25 mg/L
- Reported statistics and error estimates:
- ToxRat Solutions GmbH 2001-2010. Toxicity Response Analysis and Testing Software version 2.10
- Validity criteria fulfilled:
- yes
- Conclusions:
- The test is valid as shown by the EC50 of the reference compound 3,5-dichlorophenol (8 mg/L) and the coefficient of variation of the replicates of the control oxygen uptake rates (16%). The average oxygen uptake rate for the control replicates is lower (13 mg O2/g dry weight/hour) than the prescribed rate (>20 mg O2/g dry weight /hour). This does not influence the sensitivity of the activated sludge as shown by comparable EC50 values of the reference substance 3,5-dichlorophenol at oxygen uptake rates lower and higher than the prescribed rate (AkzoNobel 2012). The lower average oxygen uptake rate does therefore not compromise the test outcome in terms of robustness of the endpoints obtained for the test and reference substance and does not invalidate the test.
- Executive summary:
In order to predict effects of chemicals in the environment and in biological waste water treatment plants, an activated sludge respiration inhibition test was performed. The toxicity to activated sludge was determined in accordance with OECD Test Guideline 209, and in compliance with the OECD principles of Good Laboratory Practice.
The toxicity of substituted amino acid (2) solution FC-C 13587 to activated sludge was determined at a contact time of 3 hours, using various concentrations of the test substance. The inhibitory effect of substituted amino acid (2) solution FC-C 13587 at a particular concentration is expressed as a percentage of the two controls. From the results EC values were calculated. The EC10 of substituted amino acid (2) solution FC-C 13587 for activated sludge after 3 hours contact time is 591 mg/L with 95% confidence limits of 276 and 2933 mg/L. The EC20, EC50 and EC80 of substituted amino acid (2) solution FC-C 13587 for activated sludge could not be determined. Substituted amino acid (2) solution FC-C 13587 is therefore considered not harmful to activated sludge.
Reference
Table I Respiration rates of the activated sludge, inhibition percentages, pH and temperature at various concentrations of 3,5-dichlorophenol after 3 hours contact time.
Concentration (mg/L) |
Activity (mg O2/g/min) |
Inhibition (%) |
pH at start test |
pH at end test |
Temperature at end test (°C) |
Control |
0.2667 |
- |
7.1 |
7.2 |
20.2 |
Control |
0.1944 |
- |
7.0 |
7.2 |
20.2 |
Control* |
0.1905 |
- |
7.1 |
7.2 |
20.2 |
Control* |
0.1852 |
- |
7.1 |
7.2 |
20.3 |
1.27* |
0.1667 |
11 |
7.1 |
7.2 |
20.2 |
2.54 |
0.1481 |
36 |
7.1 |
7.2 |
20.2 |
5.09 |
0.1567 |
32 |
7.1 |
7.2 |
20.1 |
10.18 |
0.1037 |
55 |
7.1 |
7.2 |
20.1 |
20.36 |
0.0630 |
73 |
7.0 |
7.2 |
20.2 |
40.71 |
0.0333 |
86 |
7.0 |
7.3 |
20.0 |
pH stock solution 3,5-dichlorophenol = 6.7
pH synthetic sewage = 7.1
* = corresponding control and reference concentration
Table II Respiration rates of the activated sludge, inhibition percentages, pH and temperature at various concentrations of sodium hydrogen substituted amino acid (2) solution FC-C 13587 after 3 hours contact time.
Concentration (mg/L) |
Activity (mg O2/g/min) |
Inhibition (%) |
pH at start test |
pH at end test |
Temperature end test (°C) |
Control |
0.1833 |
- |
7.1 |
7.2 |
20.3 |
Control |
0.1905 |
- |
7.1 |
7.2 |
20.2 |
Control* |
0.1905 |
- |
7.1 |
7.2 |
20.2 |
Control* |
0.1852 |
- |
7.1 |
7.2 |
20.3 |
20.0 |
0.2083 |
-11 |
7.2 |
7.2 |
20.3 |
40.0 |
0.1917 |
-3 |
7.2 |
7.2 |
20.2 |
80.0 |
0.2083 |
-11 |
7.2 |
7.2 |
20.2 |
160.0 |
0.1963 |
-5 |
7.2 |
7.2 |
20.4 |
319.9 |
0.2000 |
-7 |
7.2 |
7.2 |
20.4 |
639.9#* |
0.1667 |
11 |
6.9 |
7.1 |
20.3 |
1279.9#* |
0.1556 |
17 |
6.7 |
7.0 |
20.2 |
Concentration stock solution sodium hydrogen substituted amino acid (2) solution FC-C 13587 = 1 g/L, pH stocksolution used = 3.5
# Concentration stock solution sodium hydrogen substituted amino acid (2) solution
FC-C 13587 = 2 g/L, pH stocksolution used = 3.4
* = corresponding control and test substance concentration
Description of key information
The toxicity of substituted amino acid (2) solution FC-C 13587 to activated sludge was determined according to OECD 209 under GLP conditions at a contact time of 3 hours, using various concentrations of the test substance. The inhibitory effect of substituted amino acid (2) solution FC-C 13587 at a particular concentration is expressed as a percentage of the two controls. From the results EC values were calculated. The EC10 of substituted amino acid (2) solution FC-C 13587 for activated sludge after 3 hours contact time is 164.3 mg a.i./L with 95% confidence limits of 76.7 and 815.4 mg/L. The EC20, EC50 and EC80 of substituted amino acid (2) solution FC-C 13587 for activated sludge could not be determined. Substituted amino acid (2) solution FC-C 13587 is therefore considered not harmful to activated sludge.
Key value for chemical safety assessment
- EC10 or NOEC for microorganisms:
- 164.3 mg/L
Additional information
Good quality data obtained with other types of microbial inhibition test methods, degradation- or sewage treatment simulation tests, can be also used to meet the REACH requirements
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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