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EC number: 946-342-2 | 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 algae and cyanobacteria
Administrative data
Link to relevant study record(s)
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 06 March 2017 to 20 April 2017
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Version / remarks:
- 2011
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- 2009
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: All test concentrations and the control (containing algae).
- Sampling method: Duplicate samples of the freshly prepared test media (containing algae) were taken. Samples were also taken after 6, 24 and 48 hours and at the end of the test (after the 72 hour test period).
- Sample storage conditions before analysis: All samples were diluted by a factor of two with acetonitrile and were stored in a freezer (≤ -20 °C), protected from light until analysis was performed. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The highest test item concentration of nominal 100 mg test item/L was prepared by mixing 93.2 mg test item into 932 mL test water. The solution was carefully shaken for 1 hour to dissolve as much of the test item as possible. After cessation of shaking and a following period (30 min) of settling to allow phase separation, the aqueous phase, i.e. the water accommodated fraction, was drawn off carefully and used as the test medium of the corresponding nominal test concentration. Adequate volumes of this water accommodated fraction were mixed into test water to obtain the desired dilutions. The test media were prepared just before introduction of the algae (= start of the test).
- Controls: Dilution water control - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Strain: KORSHIKOV, Strain No. 61.81 SAG
- Age of inoculum: Exponentially growing pre-culture
- Method of cultivation: The algae were cultivated in the laboratories under standardised conditions according to the test guidelines.
ACCLIMATION
- Acclimation period: 3 days
- Culturing media and conditions: Same as test - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Hardness:
- 0.24 mmol/L (24 mg/L) as CaCO3
- Test temperature:
- 22.2 to 23.0 °C
- pH:
- 8.1 to 8.2 at test start
9.7 to 10.1 at test end - Nominal and measured concentrations:
- Water accommodated fractions of the oversaturated stock solution of nominal 100 mg/L and dilutions of 1:3, 1:9, 1:27 and 1:81 were tested, corresponding to initial mean measured test concentrations of 7.94, 2.38, 0.761, 0.250 and 0.0586 mg test item/L.
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 50 mL Erlenmeyer flasks.
- Type: Closed
- Material, size, headspace, fill volume: 50 mL test volume.
- Initial cells density: 5000 algal cells per mL test medium.
- Control end cells density: 664910 cells/mL.
- No. of vessels per concentration: 3 (plus 3 additional replicates of each test item concentration for analytical dose verification after 6, 24 and 48 hours and 1 additional replicate of each test concentration without algae to provide a "blank" for the spectrophotometric measurements).
- No. of vessels per control: 6 (plus 3 additional replicates of each test item concentration for analytical dose verification after 6, 24 and 48 hours and 1 additional replicate of each test concentration without algae to provide a "blank" for the spectrophotometric measurements).
GROWTH MEDIUM
- Standard medium used: Yes, OECD Medium
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Deionised water
- Intervals of water quality measurement: Temperature and appearance of the test item in the test media was recorded at the start of the test and after 24, 48 and 72 hours of exposure. pH was measured at the start and end of the test.
OTHER TEST CONDITIONS
- Sterile test conditions: No.
- Adjustment of pH: The pH was adjusted with 1 M HCl to pH 8.1.
- Photoperiod: Continuous illumination.
- Light intensity and quality: 6250 to 7170 lux (mean 6688 lux).
EFFECT PARAMETERS MEASURED :
- Determination of cell concentrations: Spectrophotometer, cell density was measured after 24, 48 and 72 hours of exposure.
- Other: Algal cells were examined microscopically after 72 hours of exposure.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3
- Range finding study: Yes - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 7.94 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- > 7.94 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 7.94 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Remarks on result:
- other: 95% confidence interval: 4.03 to >7.94 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 1.75 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 2.38 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks:
- yield
- Details on results:
- - Exponential growth in the control (for algal test): Yes
- Observation of abnormalities (for algal test): The microscopic examination of the shape of the algal cells after 72 hours of test duration did not show any difference between the algae that had been growing up to the time weighted average test concentration of 7.94 mg test item/L and the algal cells in the control. Thus, the shape of the algal cells was not obviously affected up to this test concentration, the highest concentration tested.
- Colour differences: In the highest test concentration of time weighted average test concentrations of 5.95 mg test item/L, the test item caused slightly yellowish coloration only at test start. - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- EC50: 0.549 mg/L for yield, 1.12 mg/L for growth rate and 0.567 mg/L for biomass, after 72 hours exposure.
- Other: Test performed in November/December 2016, project number 110664210. - Reported statistics and error estimates:
- Based on the calculated cell densities, the 72 hour median effect concentration for growth rate (ErC50) and yield (EyC50), the corresponding 20% and 10% effect concentrations (EC20 and EC10, respectively) and where possible their 95 %-confidence limits were calculated with a 3-parametric normal distribution function. For the determination of the 72 hours LOEC and the 72 hours NOEC, the calculated growth rates and yields at each test concentration were tested for significant differences compared to the control values by Williams t-test (yield) and Step-down Jonckheere Terpstra test (growth rate). The software used to perform the statistical analysis was ToxRat Professional, Version 3.2.1, ToxRat Solutions GmbH.
- Validity criteria fulfilled:
- yes
- Remarks:
- In the controls, the 72-hour cell density increase was 133.0-fold (must be ≥ 16), the coefficient of variation (CV) of sectional (daily) growth rates was 13.1% (must be ≤ 35%) and the CV of average growth between replicates was 3.3% (must be ≤ 7%).
- Conclusions:
- The 72-hour EC50 and EC10 values for growth rate were determined to be > 7.94 mg test item/L, based on initial mean measured concentrations. The 72-hour EC50 and EC10 values for yield were calculated to be >7.94 and 1.75 mg test item/L, respectively. The 72-hour NOEC was determined to be 2.38 mg test item/L for both growth rate and yield.
- Executive summary:
The influence of the test item on the growth of the freshwater green algae (Pseudokirchneriella subcapitata) was assessed in a 72 -hour static test according to OECD Guideline 201 and EU Method C.4. Water accommodated fractions of the oversaturated stock solution of nominal 100 mg/L test item and dilutions of 1:3, 1:9, 1:27 and 1:81 were tested, corresponding to initial mean measured test concentrations of 7.94, 2.38, 0.761, 0.250 and 0.0586 mg test item/L. The test was conducted under closed conditions with a dilution water control and additional replicates of the test item solutions and control for analytical dose verification and spectrophotometric blanks. The test item and its hydrolysis product were quantified by LC-MS/MS. The 72-hour EC50 and EC10 values for growth rate were determined to be > 7.94 mg test item/L. The 72-hour EC50 and EC10 values for yield were calculated to be >7.94 and 1.75 mg test item/L, respectively. The 72-hour NOEC was determined to be 2.38 mg test item/L for both growth rate and yield. The test item concentrations decrease significantly within 24 hours, whereas the concentrations of the hydrolysis product increase significantly over all test concentrations. Since the biological inhibition was observed mainly in the first 24 hours of the test, it is suggested the inhibition is caused mainly by the test item rather than the hydrolysis products. This study is considered to be reliable without restriction (Klimisch 1) as it was conducted according to guideline with no limitations in experimental design or reporting.
Reference
Table 1. Mean Algal Cell Densities during the Test Period of 72 Hours
Initial mean measured concentration (mg test item/L) | Mean density of algal cells (10000/mL) | ||
24 hours | 48 hours | 72 hours | |
Control | 2.360 (SD 0.435) | 14.313 (SD 4.105) | 66.491 (SD 10.600) |
0.0586 | 2.133 (SD 0.151) | 17.210 (SD 2.115) | 79.849 (SD 4.827) |
0.250 | 1.277 (SD 0.087) | 14.495 (SD 2.282) | 75.529 (SD 13.200) |
0.761 | 1.881 (SD 0.231) | 13.921 (SD 3.613) | 67.116 (SD 11.819) |
2.38 | 1.478 (SD 0.349) | 10.789 (SD 1.675) | 61.830 (SD 1.805) |
7.94 | 1.378 (SD 0.151) | 8.440 (SD 1.455) | 48.757 (SD 8.061) |
SD: Standard Deviation
Table 2. Yield and Percentage Inhibition during the Test Period of 72 Hours
Initial mean measured concentration (mg test item/L) | 24 hours | 48 hours | 72 hours | |||
Mean yield (10000 cells/mL) | Inhibition (%) | Mean yield (10000 cells/mL) | Inhibition (%) | Mean yield (10000 cells/mL) | Inhibition (%) | |
Control | 1.860 (SD 0.435) | - | 13.813 (SD 4.105) | - | 65.991 (SD 10.600) | - |
0.0586 | 1.633 (SD 0.151) | 12.2 | 16.710 (SD 2.115) | -21.0 | 76.349 (SD 4.827) | -20.2 |
0.250 | 0.777 (SD 0.087) | 58.2* | 13.995 (SD 2.282) | -1.3 | 75.029 (SD 13.200) | -13.7 |
0.761 | 1.381 (SD 0.231) | 25.7* | 13.421 (SD 3.613) | 2.8 | 66.616 (SD 11.819) | -0.9 |
2.38 | 0.978 (SD 0.349) | 47.4* | 10.289 (SD 1.675) | 25.5 | 61.330 (SD 1.805) | 7.1 |
7.94 | 0.878 (SD 0.151) | 52.8* | 7.940 (SD 1.455) | 42.5* | 48.257 (SD 8.061) | 26.9* |
SD: Standard Deviation
Negative values in ‘% inhibition’ indicate an increase in growth relative to that of the control
* Mean value significantly different from the control (tested with Williams t-test, α = 0.05, one-sided)
Table 3. Growth Rate and Percentage Inhibition during the Test Period of 72 Hours
Initial mean measured concentration (mg test item/L) | 24 hours | 48 hours | 72 hours | |||
Mean growth rate (1/day) | Inhibition (%) | Mean growth rate (1/day) | Inhibition (%) | Mean growth rate (1/day) | Inhibition (%) | |
Control | 1.536 (SD 0.195) | - | 1.661 (SD 0.139) | - | 1.626 (SD 0.054) | - |
0.0586 | 1.449 (SD 0.071) | 5.7 | 1.767 (SD 0.062) | -6.4 | 1.691 (SD 0.021) | -3.9 |
0.250 | 0.936 (SD 0.067) | 39.1* | 1.679 (SD 0.082) | -1.1 | 1.669 (SD 0.060) | -2.6 |
0.761 | 1.320 (SD 0.121) | 14.1* | 1.653 (SD 0.124) | 0.5 | 1.630 (SD 0.056) | -0.2 |
2.38 | 1.063 (SD 0.257) | 30.8* | 1.532 (SD 0.080) | 7.8 | 1.606 (SD 0.010) | 1.3 |
7.94 | 1.010 (SD 0.110) | 34.3* | 1.408 (SD 0.091) | 15.2* | 1.523 (SD 0.058) | 6.3* |
SD: Standard Deviation
Negative values in ‘% inhibition’ indicate an increase in growth relative to that of the control
* Mean value significantly different from the control (tested with Step-down Jonckheere-Terpstra test, α = 0.05, one-sided)
Description of key information
The 72-hour EC50 and EC10 values for growth rate were determined to be > 7.94 mg test item/L, based on initial mean measured concentrations. The 72-hour EC50 and EC10 values for yield were calculated to be >7.94 and 1.75 mg test item/L, respectively. The 72-hour NOEC was determined to be 2.38 mg test item/L for both growth rate and yield.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 7.94 mg/L
- EC10 or NOEC for freshwater algae:
- 7.94 mg/L
Additional information
The influence of the test item on the growth of the freshwater green algae (Pseudokirchneriella subcapitata) was assessed in a 72-hour static test according to OECD Guideline 201 and EU Method C.4 (2017). Water accommodated fractions of the oversaturated stock solution of nominal 100 mg/L test item and dilutions of 1:3, 1:9, 1:27 and 1:81 were tested, corresponding to initial mean measured test concentrations of 7.94, 2.38, 0.761, 0.250 and 0.0586 mg test item/L. The test was conducted under closed conditions with a dilution water control and additional replicates of the test item solutions and control for analytical dose verification and spectrophotometric blanks. The test item and its hydrolysis product were quantified by LC-MS/MS. The 72-hour EC50 and EC10 values for growth rate were determined to be > 7.94 mg test item/L. The 72-hour EC50 and EC10 values for yield were calculated to be >7.94 and 1.75 mg test item/L, respectively. The 72-hour NOEC was determined to be 2.38 mg test item/L for both growth rate and yield. The test item concentrations decrease significantly within 24 hours, whereas the concentrations of the hydrolysis product increase significantly over all test concentrations. Since the biological inhibition was observed mainly in the first 24 hours of the test, it is suggested the inhibition is caused mainly by the test item rather than the hydrolysis products. This study is considered to be reliable without restriction (Klimisch 1) as it was conducted according to guideline with no limitations in experimental design or reporting.
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