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Reaction mass ofDisodium [1-{[2-(hydroxy-kO)-3,5-dinitrophenyl]diazenyl-kN1}naphthalen-2-olato(2-)-kO][3-(hydroxy-kO)-4-{[2-(hydroxy-kO)naphthalen-1-yl]diazenyl-kN1}-7-nitronaphthalene-1-sulfonato(3-)]chromate(2-) andDisodium [1-{[2-(hydroxy-kO)-3,5-dinitrophenyl]diazenyl-kN2}naphthalen-2-olato(2-)-kO][3-(hydroxy-kO)-4-{[2-(hydroxy-kO)naphthalen-1-yl]diazenyl-kN1}-7-nitronaphthalene-1-sulfonato(3-)]chromate(2-)
EC number: 944-038-4 | 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
Toxicity to aquatic plants other than algae
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic plants other than algae
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- Experimental start date - 30 March 2016; Experimental completion date - 12 August 2016; Study completion date - 13 October 2016.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 221 (Lemna sp. Growth Inhibition Test)
- Version / remarks:
- March 2006
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- SOURCE OF TEST MATERIAL
- Source and batch No.of test material: FAT 20037 TE; batch no.: 1501016 (China)
- Expiration date of the batch: 19 May 2020
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Approximately 4 ºC in the dark
OTHER SPECIFICS:
- Physical state/Appearance: Black solid
- Purity: 45 % - Analytical monitoring:
- yes
- Details on sampling:
- Samples were taken from the control and each test group from the bulk test preparation on Day 0 and from the pooled replicates on Day 7 for quantitative analysis. Duplicate samples were taken at each occasion and stored frozen for further analysis if necessary.
- Vehicle:
- no
- Details on test solutions:
- A nominal amount of test item (100 mg) was dissolved in culture medium and the volume adjusted to 1 liter to give a 100 mg/L test concentration from which a series of dilutions was made to give further test concentrations of 10 and 1.0 mg/L.
Each of the prepared concentrations was inverted several times to ensure adequate mixing and homogeneity. - Test organisms (species):
- Lemna minor
- Details on test organisms:
- TEST ORGANISM
- Common name: Lemna minor
- Source: Canadian Phycological Culture Centre, University of Waterloo, Ontario, Canada.
- OTHER SPECIFICS: Cultures were maintained in the laboratory by the periodic replenishment of culture medium. The culture was maintained in the laboratory at a temperature of 24 ± 1 ºC under continuous illumination (intensity approximately 7000 lux) for at least 7 days prior to the start of the test. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 7 d
- Test temperature:
- 24 ± 1 ºC
- pH:
- Control: 6.7 - 10.2,
At 1 mg/L: 6.7 - 10
3.2 mg/L: 6.7 - 8.9
10 mg/L: 6.7 - 8.3
32 mg/L: 6.7 - 7.7
100 mg/L: 6.7 - 7.5 - Nominal and measured concentrations:
- 1.0, 3.2, 10, 32 and 100 mg/L (nominal)
- Details on test conditions:
- TEST SYSTEM
- Incubation chamber used: Yes
- Test vessel: Conical flasks
- Material, size, headspace, fill volume: Glass, 500 mL.
- No. of colonies per vessel: 3 colonies
- No. of fronds: 9 fronds
- No. of vessels per concentration (replicates): three
- No. of vessels per control (replicates): three
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of frond number: The number of fronds present in each test and control culture was recorded on Days 0, 2, 5 and 7 along with observations on frond size, appearance, root length and number of colonies present.
- Determination of biomass: [dry weight] The dry weight of the fronds in each control and treatment group was determined on Day 7. At the start of the test six replicate samples of fronds identical to those used to inoculate the test vessels were taken and the dry weight determined. At the end of the test the dry weight of colonies from each control and test vessel was determined by blotting the colonies dry and drying at 60 °C to constant weight.
OTHER TEST CONDITIONS
- Adjustment of pH: The pH of the prepared culture medium will be adjusted, if necessary, to 6.5 ± 0.2 with either 1M HCl or NaOH.
- Photoperiod: Continious
- Light intensity and quality: 7000 lux.
RANGE-FINDING STUDY
- Test concentrations: nominal test concentrations of 1.0, 10 and 100 mg/L for a period of 7 days
- Results used to determine the conditions for the definitive study: The results showed no significant effect on growth at the test concentration of 1.0 mg/L. However, growth was observed to be reduced at 10 and 100 mg/L. Based on this information test concentrations of 1.0, 3.2, 10, 32 and 100 mg/L were selected for the definitive test. - Reference substance (positive control):
- yes
- Remarks:
- A positive control (Envigo Study Number MM01PC) used 3,5-dichlorophenol as the reference item at concentrations of 0.625, 1.25, 2.5, 5.0 and 10 mg/L. Exposure conditions for the positive control were similar to those in the definitive test.
- Key result
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- 92 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Duration:
- 7 d
- Dose descriptor:
- EC50
- Effect conc.:
- 5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Duration:
- 7 d
- Dose descriptor:
- NOEC
- Effect conc.:
- 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- yield
- Details on results:
- - Any visual signs of phytotoxicity (abnormalities):
- Decrease in frond size: No
- Necrosis / chlorosis: No
- Sinking of fronds: On Day 7 - fronds lying very close together, loss of buoyancy and some fronds were submerged. - Results with reference substance (positive control):
- The EC50 values for 3,5-dichlorophenol based on average specific growth rate, EC50 (frond number): 3.4 mg/L and EC50 (dry weight): 3.0 mg/L
The EC50 values based on yield, EC50 (frond number): 1.8 mg/L and EC50 (dry weight): 1.4 mg/L - Validity criteria fulfilled:
- yes
- Conclusions:
- The EC50 (frond number) based on the average specific growth rate was determined to be >100 mg/L.
- Executive summary:
A study was performed according to OECD test guideline 221 in a GLP certified laboratory, to assess the effect of the test item on the growth of the freshwater plant Lemna minor. Following a preliminary range-finding test, Lemna minor was exposed to an aqueous solution of the test item at concentrations of 1.0, 3.2, 10, 32 and 100 mg/L (three replicate flasks per concentration) for a period of 7 days, under constant illumination at a temperature of 24 ± 1 °C. The number of fronds in each control and treatment group was recorded on Days 0, 2, 5 and 7 along with observations on plant development. Analytical verification of concentration was conducted using HPLC method. Samples were drawn at the start and end of study for this purpose. Chemical analysis of the test preparations on Day 0 (fresh media) showed measured test concentrations to range from 91 % to 99 % of nominal. A concentration dependent decline in measured concentrations was observed at 72 hours in the range of 18 % of nominal at 1.0 mg/L, through to 99 % of nominal at 100 mg/L. Based on the findings of the study, the EC50 values for the test substance based on average specific growth rate were, EC50 (frond number): >100 mg/L, EC50 (dry weight): 92 mg/L, while the EC50 values based on yield were, EC50 (frond number): 10 mg/L, EC50 (dry weight): 5 mg/L.
Reference
Verification of Test Concentrations
Chemical analysis of the test preparations on Day 0 (fresh media) showed measured test concentrations to range from 91 % to 99 % of nominal. A concentration dependent decline in measured concentrations was observed at 72 hours in the range of 18 % of nominal at 1.0 mg/L, through to 99 % of nominal at 100 mg/L. Post-study stability work conducted at concentrations of 1.0 and 100 mg/L showed that the test item was stable under test conditions for a 7-Day period. As such the decline in measured concentrations observed in the definitive test was considered to be due to adsorption of the test item to the biomass present. It can therefore be considered that the lemna were exposed to nominal concentrations of the test item and as such the results have been calculated based on nominal test concentrations only.
Validation Criteria
The doubling time of the control cultures was 1.70 days in line with the OECD Guideline that states the doubling time should be less than 2.5 days:
Mean frond number in control cultures at day 0: 9
Mean frond number in control cultures at day 7: 117
Growth Data Based on Frond Number
Average Specific Growth Rate
ErC10 (frond number) = 1.9 mg/L
ErC20 (frond number) = 6.5 mg/L
ErC50 (frond number) = >100 mg/L
* Where: ErCx = the test concentration that reduced average specific growth rate by x%.
Statistical analysis of the average specific growth rate data was carried out for the control and all test concentrations using one way analysis of variance incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf 1981) and Dunnett's multiple comparison procedure for comparing several treatments with a control (Dunnett 1955). There were no statistically significant differences between the control and 1.0 mg/L test concentration (P≥0.05), however all other test concentrations were significantly different ( P<0.05) and, therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of average specific growth rates calculated from frond numbers was 1.0 mg/L. Correspondingly the “Lowest Observed Effect Concentration” (LOEC) was determined to be 3.2 mg/L.
Yield
EyC10 (frond number) = 1.1 mg/L
EyC20 (frond number) = 1.6 mg/L
EyC50 (frond number) = 10 mg/L
Where: EyCx = the test concentration that reduced yield by x%.
Statistical analysis of the yield data was carried out. There were no statistically significant differences (P≥0.05), between the control and 1.0 mg/L test concentration (P≥0.05), however all other test concentrations were significantly different (P<0.05) and, therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of yield calculated from frond numbers was 1.0 mg/L. Correspondingly the “Lowest Observed Effect Concentration” (LOEC) was determined to be 3.2 mg/L.
Growth Data Based on Dry Weight
Average Specific Growth Rate
ErC10 (dry weight) = 0.67 mg/L
ErC20 (dry weight) = 4.1 mg/L
ErC50 (dry weight) = 92 mg/L
Where: ErCx = the test concentration that reduced average specific growth rate by x%.
Statistical analysis of the average specific growth rate data was carried out. There were no statistically significant differences (P≥0.05), between the control and 1.0 mg/L test concentration (P≥0.05), however all other test concentrations were significantly different (P<0.05) and, therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of average specific growth rate calculated from dry weight was 1.0 mg/L. Correspondingly the “Lowest Observed Effect Concentration” (LOEC) was determined to be 3.2 mg/L.
Yield
EyC10 (dry weight) = 1.1 mg/L
EyC20 (dry weight) = 1.4 mg/L
EyC50 (dry weight) = 5.0 mg/L
Where: EyCx = the test concentration that reduced yield by x%.
Statistical analysis of the yield data was carried out. There were no statistically significant differences (P≥0.05), between the control and 1.0 mg/L test concentration (P≥0.05), however all other test concentrations were significantly different (P<0.05) and, therefore the "No Observed Effect Concentration" (NOEC) in terms of inhibition of yield calculated from dry weight was 1.0 mg/L. Correspondingly the “Lowest Observed Effect Concentration” (LOEC) was determined to be 3.2 mg/L.
Description of key information
A study was performed according to OECD test guideline 221 in a GLP certified laboratory, to assess the effect of the test item on the growth of the freshwater plant Lemna minor. Following a preliminary range-finding test, Lemna minor was exposed to an aqueous solution of the test item at concentrations of 1.0, 3.2, 10, 32 and 100 mg/L (three replicate flasks per concentration) for a period of 7 days, under constant illumination at a temperature of 24 ± 1 °C. The number of fronds in each control and treatment group was recorded on Days 0, 2, 5 and 7 along with observations on plant development. Analytical verification of concentration was conducted using HPLC method. Samples were drawn at the start and end of study for this purpose. Chemical analysis of the test preparations on Day 0 (fresh media) showed measured test concentrations to range from 91% to 99% of nominal. A concentration dependent decline in measured concentrations was observed at 72 hours in the range of 18 % of nominal at 1.0 mg/L, through to 99 % of nominal at 100 mg/L. Based on the findings of the study, the EC50 values for the test substance based on average specific growth rate were, EC50 (frond number): >100 mg/L, EC50 (dry weight): 92 mg/L, while the EC50 values based on yield were, EC50 (frond number): 10 mg/L, EC50 (dry weight): 5 mg/L.
Key value for chemical safety assessment
- EC50 for freshwater plants:
- 100 mg/L
Additional information
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