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EC number: 200-796-1 | CAS number: 73-24-5
- 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:
- 2018-09-17 to 2018-09-20
- 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)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- - Sampling method: At the start of the exposure (0 hours), samples of the fresh media were taken after preparation of each concentration and analyzed. At the end of the exposure (72 hours), samples of the old media were taken from the test vessels for analyses.
- Sample storage conditions before analysis: If necessary, all original samples were stored at room temperature before preparation. Prepared samples were stored in an autosampler at room temperature until analysis. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A stock solution with a nominal concentration of 100 mg/L of the test item was freshly prepared prior to the start of the exposure. An appropriate amount of the test item was weighed out and transferred with an appropriate amount of dilution water into the glass flask. This dispersion was treated with ultrasound for 30 minutes at room temperature. Care was taken to avoid bacterial contamination. The test solutions were prepared by diluting the stock solution with dilution water.
- Controls: Six replicates (without test item) were tested under the same test conditions as the test item replicates. Potassium dichromate was tested twice per year as a reference item.
- Evidence of undissolved material: No - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Strain: HINDAK, CCAP 278/4
- Synonyms: Selenastrum capricornutum; Ankistrodesmus subcapitata; Raphidocelis subcapitata; Ankistrodesmus bibraianus
- Source: Culture Collection of Algae and Protozoa (CCAP), SAMS Research Services Ltd
Dunstaffnage Marine Laboratory, Dunbeg, OBAN; Argyll PA37 1QA; Scotland, UK
- Method of cultivation: Fresh stocks are prepared every month on Z-Agar. Light intensity amounted to 2590 - 5180 lux per day. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- No
- Hardness:
- Nominal hardness of medium: 0.24 mmol Ca+Mg/L.
- Test temperature:
- 21 - 24°C ± 2°C
- pH:
- 7.74 - 9.31
- Dissolved oxygen:
- Not measured
- Salinity:
- Not applicable
- Conductivity:
- Not measured
- Nominal and measured concentrations:
- Nominal: 0.768,1.92, 4.80, 12.0, 30.0 mg/L
Mean measured: 2.06, 4.71, 12.1, 28.9 mg/L (fresh media, 0 hours); 1.68, 4.84, 12.0, 29.9 mg/L (old media, 72 hours) - Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Material, size, headspace, fill volume: Glass, nominal volume: 250 mL, fill volume: 100 mL, sealed with cotton wool plugs
- Aeration: Test containers were placed on a rotary shaker and oscillated at approximately 70 rpm.
- Initial cells density: 6442 cells/mL
- Control end cells density: 2084121 cells/mL (mean)
- No. of vessels per concentration: 3
- No. of vessels per blank control: 6
GROWTH MEDIUM
- Standard medium used: Yes
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Ultra-pure water used for preparation of test medium
OTHER TEST CONDITIONS
- Sterile test conditions: Yes
- Adjustment of pH: No
- Photoperiod: Continuously illuminated
- Light intensity: 2590 - 5180 lux
- Light homogeneity (target): Within ± 15% over incubation area
EFFECT PARAMETERS MEASURED
- Determination of cell concentrations: Chlorophyll a-fluorescence
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5
- Range finding study: Non-GLP preliminary test performed
- Test concentrations: 1.00, 10.0, 100 mg/L.
- Results used to determine the conditions for the definitive study: Yes - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 4.33 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95% confidence interval: 3.57 - 4.56 mg/L
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 2.82 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95% confidence interval: 2.20 - 3.70
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 3.29 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95% confidence interval: 2.58 - 3.92 mg/L
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 1.92 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 4.8 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 3.93 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95% confidence interval: 2.23-6.65 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 3.63 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95% confidence interval: 2.23-6.65 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 3.63 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95% confidence intervals: 2.11 -6.45 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 3.43 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95% confidence interval: 2.02 - 6.40 mg/L
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 1.92 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 4.8 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Details on results:
- - Exponential growth in the control (for algal test): Yes
- Observation of abnormalities: No
- Unusual cell shape: No
- Colour differences: No
- Flocculation: No
- Adherence to test vessels: No
- Aggregation of algal cells: No
- Any stimulation of growth found in any treatment: No
- Any observations that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- EC50: 1.09 mg/L (growth rate), 0.548 mg/L (yield) - Reported statistics and error estimates:
- Shapiro-Wilk's test on normal distribution was calculated with a significance level of 0.01.
Levene's test on variance homogeneity was calculated with a significance level of 0.01.
Monotonicity of Concentration/Response was calculated by Trend Analysis by Contrasts (significance level 0.05) for yield.
Monotonicity of Concentration/Response was calculated by non-parametric trend analysis by contrasts (significance level 0.05) for growth rate.
SD Jonckheere-Terpsta test procedure was done with a significance level 0.05. - Validity criteria fulfilled:
- yes
- Conclusions:
- In a growth inhibition test with Raphidocelis subcapitata according to OECD TG 201, EU Method C.3 and GLP principles, the 72-hour ErC50 was 4.33 mg/L and the EyC50 was 3.93 mg/L (nominal). The 72-hour ErC10 and EyC10 was determined to be 2.82 mg/L and 3.43 mg/L (nominal), respectively.
- Executive summary:
The toxicity of the test item to the unicellular freshwater green alga Raphidocelis subcapitata was determined according to OECD TG 201, EU Method C.3 and the principles of GLP. The study was conducted under static conditions with an initial cell density of 6442 cells/mL. Five concentrations were tested in a geometrical series with factor of 2.5: 0.768, 1.92, 4.80, 12.0, 30.0 mg/L. Three replicates were tested for each test item concentration and six replicates for the control. The test item concentrations in the test media were confirmed by HPLC-DAD analysis to be within ± 20% of the nominal concentrations. Therefore, all effect values are based on the nominal test item concentrations. In result, the 72-hour ErC50 was 4.33 mg/L and the EyC50 was 3.93 mg/L. The 72-hour ErC10 and EyC10 was determined to be 2.82 mg/L and 3.43 mg/L (nominal), respectively. All validity criteria of the guidelines were fulfilled.
Reference
Table 1: Cell Densities
Nominal test item concentration[mg/L] | Replicate | Cell density [cells/mL] | |||
No. | 0 hours | 24 hours | 48 hours | 72 hours | |
30 | 1 | 6442 | 8224 | 9376 | 7509 |
2 | 6442 | 9811 | 10409 | 7284 | |
3 | 6442 | 7842 | 10824 | 7909 | |
Mean | 6442 | 8626 | 10203 | 7567 | |
12 | 1 | 6442 | 11573 | 14426 | 11501 |
2 | 6442 | 9495 | 14262 | 11258 | |
3 | 6442 | 7256 | 10992 | 11501 | |
Mean | 6442 | 9441 | 13227 | 11420 | |
4.8 | 1 | 6442 | 15270 | 33819 | 48545 |
2 | 6442 | 16799 | 42724 | 75540 | |
3 | 6442 | 13686 | 30547 | 49670 | |
Mean | 6442 | 15252 | 35697 | 57918 | |
1.92 | 1 | 6442 | 25880 | 244618 | 2035762 |
2 | 6442 | 20220 | 200966 | 2030648 | |
3 | 6442 | 26401 | 190162 | 2017242 | |
Mean | 6442 | 24167 | 211915 | 2027884 | |
0.768 | 1 | 6442 | 27481 | 221451 | 2003986 |
2 | 6442 | 23591 | 259295 | 1909998 | |
3 | 6442 | 25676 | 256564 | 2084617 | |
Mean | 6442 | 25583 | 245770 | 1999534 | |
Control | 1 | 6442 | 24224 | 201641 | 2118181 |
2 | 6442 | 30912 | 249365 | 1772964 | |
3 | 6442 | 27652 | 288737 | 2046436 | |
4 | 6442 | 27834 | 300832 | 2057061 | |
5 | 6442 | 24388 | 292927 | 2132877 | |
6 | 6442 | 20250 | 209565 | 2377205 | |
Mean | 6442 | 25877 | 257178 | 2084121 |
Table 2: Evaluation after 72 hours. Statistically significant differences of growth rates and yield compared to control values are marked (+), not significant differences are marked (-).
Nominal test item concentration [mg/L] | Replicate No. | Growth Rate [d-1] | Inhibition of Growth Rate [%] | Yield [cells/mL] | Inhibition of Yield [%] |
30.0 | 1 | 0.051 | 97 | 1067 | 100 |
2 | 0.041 | 98 | 842 | 100 | |
3 | 0.068 | 96 | 1467 | 100 | |
Mean | (+) 0.053 | 97 | 1125 | 100 | |
12.0 | 1 | 0.193 | 90 | 5059 | 100 |
2 | 0.186 | 90 | 4816 | 100 | |
3 | 0.193 | 90 | 5059 | 100 | |
Mean | (+) 0.191 | 90 | (+) 4978 | 100 | |
4.80 | 1 | 0.67 | 65 | 42103 | 98 |
2 | 0.82 | 57 | 69098 | 97 | |
3 | 0.68 | 65 | 43228 | 98 | |
Mean | (+) 0.73 | 62 | (+) 51476 | 98 | |
1.92 | 1 | 1.92 | 0 | 2029320 | 2 |
2 | 1.92 | 0 | 2024206 | 3 | |
3 | 1.92 | 1 | 2010800 | 3 | |
Mean | (-) 1.92 | 0 | (-) 2021442 | 3 | |
0.768 | 1 | 1.91 | 1 | 1997544 | 4 |
2 | 1.9 | 1 | 1903556 | 8 | |
3 | 1.93 | 0 | 2078175 | 0 | |
Mean | (-) 1.91 | 1 | (-)1993992 | 4 | |
Control | 1 | 1.93 | 2111739 | ||
2 | 1.87 | 1766522 | |||
3 | 1.92 | 2039994 | |||
4 | 1.92 | 2050619 | |||
5 | 1.93 | 2126435 | |||
6 | 1.97 | 2370763 | |||
Mean | 1.93 | 2077679 |
Table 3: Measured Concentration of the test item in the Definitive Test with Algae
Sampling date | Fresh Media, 0 hours | Old Media, 72 hours | ||
Nominal test item concentration [mg/L] | Meas. conc. [mg/L] | % | Meas. conce. [mg/L] | % |
30 | 28.9 | 96 | 29.9 | 100 |
12 | 12.1 | 101 | 12 | 100 |
4.8 | 4.71 | 98 | 4.84 | 101 |
1.92 | 2.06 | 107 | 1.68 * | 88 |
0.768 | < LOQ | < LOQ | ||
Control | < LOQ | < LOQ |
Meas. conc. =
measured concentration of the test item, dilution factors taken into
account
% =
percentage of the nominal measured concentration of the test item
LOQ = limit of quantification (1.80 mg/L of the test item)
*) = > 70% LOQ
Description of key information
In a growth inhibition test with Raphidocelis subcapitata according to OECD TG 201, the 72-hour ErC50 was 4.33 mg/L and the EyC50 was 3.93 mg/L (nominal). The 72-hour ErC10 and EyC10 was determined to be 2.82 mg/L and 3.43 mg/L (nominal), respectively (reference 6.1.5-1).
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 4.33 mg/L
- EC10 or NOEC for freshwater algae:
- 2.82 mg/L
Additional information
The toxicity of the test item to the unicellular freshwater green alga Raphidocelis subcapitata was determined according to OECD TG 201, EU Method C.3 and the principles of GLP. The study was conducted under static conditions with an initial cell density of 6442 cells/mL. Five concentrations were tested in a geometrical series with factor of 2.5: 0.768, 1.92, 4.80, 12.0, 30.0 mg/L. Three replicates were tested for each test item concentration and six replicates for the control. The test item concentrations in the test media were confirmed by HPLC-DAD analysis to be within ± 20% of the nominal concentrations. Therefore, all effect values are based on the nominal test item concentrations. In result, the 72-hour ErC50 was 4.33 mg/L and the EyC50 was 3.93 mg/L. The 72-hour ErC10 and EyC10 was determined to be 2.82 mg/L and 3.43 mg/L (nominal), respectively. All validity criteria of the guidelines were fulfilled (reference 6.1.5-1).
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