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EC number: 274-700-1 | CAS number: 70616-89-6
- 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:
- 2016
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)
- Version / remarks:
- published in O.J. L 54/1 (2016)
- Deviations:
- yes
- Remarks:
- During the measuring of oxygen consumption of abiotic control the temperature was out of prescribed range 0.1°C. This increase of temperature was minimal and lasted only in last minute of the test. The deviation has not an influence to the study results.
- 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 activated biological sludge containing mixed culture of microorganisms obtained from the second step of sewage treatment plant of Pardubice was used for the testing. The wastewater processed by the sewage treatment plant is predominantly municipal.The activated sludge was collected two days before the day of testing. After the sample collection the sludge was washed with potable water for 30 minutes and subsequently decanted for 30 minutes. This procedure was repeated three times in total. Further the sludge was fed with the 100 mL synthetic sewage feed per 2 L of diluted sludge suspension at permanent aeration till the next day. The next day the sludge was washed with potable water and decanted according to the procedures described for the first day. This procedure was repeated three times in total again.The suspended solids concentration was determined from 10 mL of sludge suspension after the last 30-minutes sedimentation. The sludge was fed with the 100 mL synthetic sewage feed per 2 L of diluted sludge suspension at permanent aeration till day of the test.Before the test the sludge was suspended in water up to concentration about 3000 mg of suspended solids per litre. The pH adjustment to 6.0 was carried out. In that way modified sludge suspension was aerated until the use.The activated sludge obtained is in conformity with the recommendations of the test guidelines.
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 3 h
- Test temperature:
- 20 ± 2°C
- pH:
- 7.08 - 7.98
- Dissolved oxygen:
- 5.92 - 9.09 mg/L at the start of the test
- Nominal and measured concentrations:
- Nominal: 10, 100 and 1000 mg/L
- Details on test conditions:
- SOLUTIONS AND CHEMICALSThe stock solution of synthetic sewage feed:Peptone: 16 gmeat extract: 11 gurea: 3 gNaCl: 0.7 gCaCl2·2H2O: 0.4 gMgSO4·7H2O: 0.2 gK2HPO4: 2.8 gdeionized water up to volume of 1000 mLThe pH value of this solution is: 7.0 - 8.0.The synthetic sewage feed for sludge adaptation was always prepared fresh on the day of the use.The solutions for dissolution and pH adjustmentH2SO4 0.5 mol/LNaOH 1.0 mol/LSterilizing solutionMercury chloride (HgCl2), of concentration of 5 g/L. 1 mL of this solution per 500 mL of reaction mixture was used for elimination of microbial activity.Specific inhibitor of nitrificationN-allylthiourea (ATU), of concentration of 2.5 g/L. 2.32 mL of this solution per 500 mL of reaction mixture was used for inhibition of nitrification in a activated sludge.INSTRUMENTS AND EQUIPMENToxygen vessels (BOD bottles) with volume of about 280 mL for measurement of oxygen concentration graduated cylinders of volume 1000 mL analytical balance AG135 (Mettler Toledo)analytical balance Kern 870-15oximeter WTW Inolab OXI 730 with membrane probe CellOx 325 and with printer stirring adapter for electrodepH meter WTW- pH 539 with pH-electrode WTW SenTix 81 with temperature probemagnetic stirrersaerated apparatus with glass ends (pressured air is passed through filter for odour and oil removal)air-conditioned room, free of dust, with automatic temperature recordingcommon laboratory equipment (pipettes, beakers, volumetric cylinders, volumetric flasks and further ones) PREPARATION OF SOLUTIONS FOR THE TEST Test substance The test substance is soluble in deionized water in concentrations needed for the test. The stock solution was prepared by dissolution of 5.0 g of the test substance in 1000 mL deionized water on the day of testing. The concentrations 10, 100 and three replicates of 1000 mg/L (with pH adjustment) and three replicates of 1000 mg/L (without pH adjustment) of the test substance were tested in the preliminary test. Reference substance The stock solution of the reference substance was prepared by dissolution of 0.1 g of 3,5-dichlorophenol in 200 mL deionized water. The ultrasound for 60 minutes to accelerate the dissolution was used. The resulting solution has a concentration of 500 mg/L. The stock solution was prepared on the day of testing. The concentration range used was 5 - 20 mg/L. It was calculated by geometric progression with factor of 2.0.TEST CONDITIONSTemperature: 20 ± 2°CExposition time: 3 hours Lighting: daily lightStirring and aeration: with filtered pressured air The pH value of reaction mixtures during the preliminary test: approx. 7 – 8 TEST PROCEDUREExperimental designThe respiration rates of the activated sludge in standard nutrient medium were measured in the presence of three concentrations of the test substance after contact time of 3 hours. The preliminary test was performed using 3 concentrations 10, 100 and 1000 mg/L. Further two blank controls, the three abiotic controls with the highest concentration of the test substance and two beakers with specific nitrification inhibitor to determine the nitrification potential of sludge were included.3,5-dichlorophenol was used as reference substance. Three concentrations in geometric progression with a factor of 2.0 (5, 10 and 20 mg/L) were selected.The inhibition effects of the test and reference substance at each concentration were expressed as the percentage of respiration rate from the average of control respiration rates determined at the beginning and the end of the test.The low level of inhibition of oxygen consumption by the test substance in the preliminary test demonstrated that a definitive test is unnecessary. Preparing of the test mixturesTest/reference substance100 mL of deionized water, the defined dose of the test or reference substance, 16 mL of the synthetic sewage feed and 250 mL of the prepared the sludge suspension were introduced into each test container. The volume was replenished up to 500 mL with deionized water.Blank controlThe blank controls contained only 16 mL of synthetic sewage feed and 250 ml of the sludge suspension and replenished up to 500 mL with deionized water.Abiotic controlThe check of the abiotic decomposition was prepared at the highest concentration of the test substance, 16 mL synthetic sewage feed without the sludge suspension and with addition of 1 mL of sterilizing solution (mercury chloride solution). The volume was replenished up to 500 mL with deionized water.Nitrification potential of sludgeThe mixtures to determine the nitrification potential of sludge included 16 mL of the synthetic sewage feed, 250 mL the sludge suspension and 2.32 mL of specific inhibitor of nitrification (N-allylthiourea solution). The volume was replenished up to 500 mL with deionized water.The activated sludge concentration in all test, reference and blank (but not abiotic control) mixtures is nominally 1.5 g/L of suspended solid.The test mixtures were prepared in turns in 15 minutes time intervals and aerated/incubated at 20 °C ± 2 °C for 3 hours. The aeration was carried out by means of glass pipettes.The pH adjustment of the test mixtures in the preliminary test was not carried out.MeasurementThe pH was determined in the reaction mixtures at the beginning of the test.The measurement of the oxygen content was performed in aliquots of the reaction mixtures after 3-hour exposure period. Bottles with narrow neck and volume of about 280 mL were filled with the sample and positioned on a magnetic stirrer. Then, the oxygen electrode (equipped with a magnetic stirring wheel) was inserted in a way that no air remained in the bottle and that the electrode sealed the bottleneck to avoid contact of the sample with the atmosphere. The decline of oxygen concentration was measured and recorded in 30-seconds intervals for 11 minutes. The pH was determined in the remaining part of the reaction mixture.
- Reference substance (positive control):
- yes
- Remarks:
- 3,5-dichlorophenol (CAS No.: 591-35-5)
- 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:
- see Table 1
- Results with reference substance (positive control):
- 3,5-dichlorophenol: EC50 = 7.9 mg/L
- Validity criteria fulfilled:
- yes
- Conclusions:
- The highest inhibition the respiration rate of the activated sludge caused by the test substance Reactive Orange 13 was 11.7 % in the preliminary test. This low level of inhibition of oxygen consumption by the test substance in the preliminary test demonstrated that a definitive test is unnecessary. The result could be used as final one, because preliminary test design includes three replicates required by guidelines. Exact value of EC50 could not be calculated, the value of EC50 is given in the form of a range:EC50 > 1 000 mg/LThe abiotic decomposition was not detected even at the highest concentration of the test substance. The nitrification respiration rate 14.6 % of the total respiration rate in the blank controls was detected in the preliminary test. In the case of observing the inhibitory effect of the test substance on the respiration of activated sludge it would be necessary to take into account the activity of nitrifying. In this case, the inhibitory effect of the test substance is only 11.7 %. This low level of inhibition demonstrated that further experiments are not needed.
- Executive summary:
The influence of the test substance, Reactive Orange 13, on the respiration rate of activated sludge was investigated after a contact time of 3 hours. Test was performed according to OECD Test Guideline No. 209 – Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation). Adopted July 22, 2010
Method C.11 – Activated Sludge Respiration Inhibition Test (Carbon and Ammonium Oxidation), Council Regulation (EU) No. 2016/266, published in O.J. L 54/1, 2016.
The preliminary test was performed using 3 concentrations 10, 100 and 1000 mg/L. Further two blank controls, the three abiotic controls with the highest concentration of the test substance was included and two beakers with specific nitrification inhibitor to determine the nitrification potential of sludge.
3,5-dichlorophenol was used as reference substance. Three concentrations in geometric progression with a factor of 2.0 (5, 10 and 20 mg/L) were selected.
The abiotic decomposition was not detected even at the highest concentration of the test substance.
The nitrification respiration rate 14.6 % of the total respiration rate in the blank controls was detected in the preliminary test. In the case of observing the inhibitory effect of the test substance on the respiration of activated sludge it would be necessary to take into account the activity of nitrifying. In this case, the inhibitory effect of the test substance is only 11.7 %. This low level of inhibition demonstrated that further experiments are not needed.
The prescribed validity criteria in the test were fulfilled. Since all criteria of acceptability were met, this study is considered to be valid.
The highest inhibition the respiration rate of the activated sludge caused by the test substance, Reactive Orange 13, was 11.7 % in the preliminary test. This low level of inhibition of oxygen consumption by the test substance in the preliminary test demonstrated that a definitive test is unnecessary. The result could be used as final one, because preliminary test design includes three replicates required by guidelines. Exact value of EC50 could not be calculated, the value of EC50 is given in the form of a range:
EC50 > 1000 mg/L
- Endpoint:
- toxicity to microorganisms, other
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1983
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with national standard methods with acceptable restrictions
- Justification for type of information:
- None
- Qualifier:
- according to guideline
- Guideline:
- other: Internal standard method
- Principles of method if other than guideline:
- None
- GLP compliance:
- no
- Specific details on test material used for the study:
- None
- Analytical monitoring:
- not specified
- Details on sampling:
- None
- Vehicle:
- not specified
- Details on test solutions:
- None
- Test organisms (species):
- other: not specified
- Details on inoculum:
- None
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 21 d
- Post exposure observation period:
- None
- Hardness:
- None
- Test temperature:
- None
- pH:
- None
- Dissolved oxygen:
- None
- Salinity:
- None
- Conductivity:
- None
- Nominal and measured concentrations:
- None
- Details on test conditions:
- None
- Reference substance (positive control):
- not specified
- Key result
- Duration:
- 21 d
- Dose descriptor:
- IC50
- Effect conc.:
- > 300 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth inhibition
- Details on results:
- None
- Results with reference substance (positive control):
- None
- Reported statistics and error estimates:
- None
- Validity criteria fulfilled:
- no
- Conclusions:
- The IC50 for FAT 40170/B was found to be greater than 300 mg/L.
- Executive summary:
The toxicity of FAT 40170/B to activated sludge was determined in a 21 days respiration inhibition test according to an internal SOP. Based on the findings of the study, the IC50 of the test substance FAT 40170/B is greater than 300 mg/L.
- Endpoint:
- toxicity to microorganisms, other
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Study period:
- 1983
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- test procedure in accordance with national standard methods with acceptable restrictions
- Justification for type of information:
- None
- Qualifier:
- according to guideline
- Guideline:
- other: Internal standard method
- Principles of method if other than guideline:
- None
- GLP compliance:
- no
- Specific details on test material used for the study:
- None
- Analytical monitoring:
- not specified
- Details on sampling:
- None
- Vehicle:
- not specified
- Details on test solutions:
- None
- Test organisms (species):
- other: not specified
- Details on inoculum:
- None
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 21 d
- Post exposure observation period:
- None
- Hardness:
- None
- Test temperature:
- None
- pH:
- None
- Dissolved oxygen:
- None
- Salinity:
- None
- Conductivity:
- None
- Nominal and measured concentrations:
- None
- Details on test conditions:
- None
- Reference substance (positive control):
- not specified
- Key result
- Duration:
- 21 d
- Dose descriptor:
- IC50
- Effect conc.:
- > 300 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth inhibition
- Details on results:
- None
- Results with reference substance (positive control):
- None
- Reported statistics and error estimates:
- None
- Validity criteria fulfilled:
- no
- Conclusions:
- The IC50 for FAT 40170/A was found to be greater than 300 mg/L.
- Executive summary:
The toxicity of FAT 40170/A to activated sludge was determined in a 21 days respiration inhibition test according to an internal SOP. Based on the findings of the study, the IC50 of the test substance FAT 40170/A is greater than 300 mg/L.
Referenceopen allclose all
Table 1: Evaluation of inhibition of respiration rates
Container number | Label | Concentration mg/L | Total inhibition |
1 | K1 | 0 | - |
2 | Z1 | 10 | - 8.3 |
3 | Z2 | 100 | - 7.6 |
4 | Z3 | 1000 | - 1.6 |
5 | Z4 | 1000 | 11.7 |
6 | Z5 | 1000 | 8.5 |
7 | R1 | 5 | 26.3 |
8 | R2 | 10 | 67.3 |
9 | R3 | 20 | 86.5 |
10 | Zab1 | 1000 | - |
11 | Zab2 | 1000 | - |
12 | Zab3 | 1000 | - |
13 | KN1 | 0 | - |
14 | KN2 | 0 | - |
15 | K2 | 0 | - |
16 | Z6 | 1000 | 20.9 |
17 | Z7 | 1000 | 23.4 |
18 | Z8 | 1000 | 19.2 |
K1, K2 – control experiment
Z1 – Z5 – test substance with pH adjustment
Z6 – Z8 – test substance without pH adjustment
R1 – R3 – reference substance
Zab1 – Zab3 – test substance – abiotic decomposition
KN1 – KN2 – nitrification potential of sludge
None
None
Description of key information
Key value for chemical safety assessment
- EC50 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.
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