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EC number: 207-396-6 | CAS number: 467-63-0
- 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
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- Nanomaterial pour density
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- 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 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
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- Experimental test result performed using standard OECD test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- GLP compliance:
- no
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: 0, 0.2, 0.54, 1.46, 3.94 and 10.64 mg/L
- Sample storage conditions before analysis: Test samples were analysed immediately after sampling - Vehicle:
- no
- Details on test solutions:
- The test solution was prepared by dissolving 1000 mg of test chemical in 1000 ml of OECD media, and allowed for stirring for 96 hours, which was then filtered and the final stock solution obtained was 1000 mg/L, verified analytically by UV-Vis Spectrophotometer. The final solubility value obtained after analytical detection is 13.71 mg/L. To have a better growth and visibility of cells, the initial cell count of the culture was kept 10,000 cells/mL.
- Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: green alga
- Length: 8 – 14 μm
- Weight: 2 - 3 μm
- Source: Sterile, unicellular, suspension cultures of algae were obtained from Denmark Technical University, Denmark and are maintained at Unique Ecotox Research Laboratory, Nagpur.
- Method of cultivation: OECD medium
ACCLIMATION
- Culturing media and conditions (same as test or not): The medium to be used for the growth of algae was OECD medium.
- Any deformed or abnormal cells observed: no - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Test temperature:
- 21 to 24 ± 2°C
- Nominal and measured concentrations:
- Test chemical concentrations used for the study were 0, 0.2, 0.54, 1.46, 3.94 and 10.64 mg/L (factor of 2.7), respectively.
- Details on test conditions:
- TEST SYSTEM
- Test vessel: Conical flasks
- Material, size, headspace, fill volume: 100 ml conical flasks filled with 60 ml was used for the study.
- Initial cells density: 10000cells/ml
- No. of organisms per vessel: 10000cells/ml
- No. of vessels per concentration (replicates): Three replicates for each test concentration
- No. of vessels per control (replicates): Three replicates for Control
GROWTH MEDIUM
- Standard medium used: yes, OECD medium was used as a test medium in the study.
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Photoperiod: 16 Hour Light Period : 8 Hour Dark Period
- Light intensity and quality: continuous, uniform fluorescent illumination(6000-10000Lux)
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Cell counts were measured using microscope.
- Chlorophyll measurement: No data
- Other: The cultures were observed daily with the help of a microscope to verify a normal and healthy appearance of the algal culture and also to observe any abnormal appearance of the algae (as may be caused by the exposure of the test item). Apart from this, the cell count of each test vessel was also noted with the help of a microscope and haemocytometer.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: All the six concentrations were in geometric series spaced by a factor of 2.7.
- Test concentrations: test concentration were: 0, 0.2, 0.54, 1.46, 3.94 and 10.64 mg/L (Nominal concentrations)
- Results used to determine the conditions for the definitive study: Mortality of test organisms
Other:
Incubation :
1. The temperature of the orbital shaking incubator was kept constant throughout the period of exposure of the experiment. The temperature was maintained at 22 ° C±2°C.
2. The test vessels were incubated with a continuous, uniform fluorescent illumination (1500Lux).
3. The pH of the control cultures needs to be noted during the study and the pH of the control medium should not increase by more than 1.5 units during the test.
4. The orbital shaking incubator was set at a speed of 120 revolutions per minute throughout the study period. This is to provide constant shaking to the algal cells to keep them in suspension and to ensure that they do not settle down on the bottom of the test vessel.
5. Study duration : The experimental phase of the study was lasted for a period of 72 hours. - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate (K2Cr2O7) was used as a reference substance.
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 0.52 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 0.47 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- The microscopic observations were also noted in each of the experimental flasks. All the cells appeared healthy, sickle shaped and green throughout the test duration in the control.
- Results with reference substance (positive control):
- - Results with reference substance valid?
- EC50: 0.868 mg/l - Reported statistics and error estimates:
- To obtain a quantitative concentration-response relationship by regression analysis, a linearizing transformation of the response data into probit was performed. Using the same, effective concentration (EC) was determined.
- Validity criteria fulfilled:
- yes
- Remarks:
- * The average increase in cell count in control group is >16 fold. * The coefficent of variation between replicates was <7% (i.e., reported to be 1.31%) * coefficent of variation in the control % CV of Average specific <35% (i.e., reported to be 4.44%).
- Conclusions:
- Based on the growth inhibition of green alga Pseudokirchneriella subcapitata by the test chemical, the 72 hr median effect concentration (ErC50) value was determined to be 0.52 mg/l (nominal conc.) and 0.47 mg/l (measured conc.), respectively.
- Executive summary:
A freshwater algal growth inhibition test was conducted for 72 hrs for assessing the effect of test chemical on green algae Pseudokirchneriella subcapitata. The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. OECD medium composed of macronutrients, micronutrients, alkaline EDTA solution and iron solution was used as a growth medium. The test solution was prepared by dissolving 1000 mg of test chemical in 1000 ml of OECD media, and allowed for stirring for 96 hours, which was then filtered and the final stock solution obtained was 1000 mg/L, verified analytically by UV-Vis Spectrophotometer. The final solubility value obtained after analytical detection is 13.71 mg/L. To have a better growth and visibility of cells, the initial cell count of the culture was kept 10,000 cells/mL. Further, exposure concentrations of 0, 0.2, 0.54, 1.46, 3.94 and 10.64 mg/L (factor of 2.7), respectively was from the stock test concentrations. Test vessel were placed in orbital shaking incubator for 72 hrs at a temperature of 21 to 24 ± 2°C and with a continuous uniform illumination of 6000 -10000 lux light intensity, respectively. The speed of the orbital shaking incubator was set at a 120 revolutions per minute throughout the study period. The cultures were counted and observed daily with the help of a microscope. Potassium dichromate (K2Cr2O7) was used as a reference substance. The 72 hr EC50 value of the reference substance was determined to be 0.868 mg/l. The biomass in the control vessel have increased exponentially by a factor of 16, the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% (i.e., reported to be 1.31%) and the mean coefficient of variation of specific growth rate was not exceeded 35% (i.e., reported to be 4.44%), respectively thus fulfilling the validity criterion. As the concentration of the test chemical being tested has not been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on measured concentration. On the basis of growth rate of the test organism, the 72 hr median effect concentration (ErC50) value was determined to be 0.52 mg/l (nominal conc.) and 0.47 mg/l (measured conc.), respectively. Thus, test chemical was considered as toxic to aquatic algae at environmental relevant concentrations and hence, considered to be classified in 'aquatic acute/chronic category 1' as per the CLP classification criteria.
Reference
Sr. No |
Concentrations mg/L |
Wavelength (nm) |
Absorbance |
Temperature (°C) |
1 |
blank |
595 |
0.00 |
25°C |
2 |
0.10 |
595 |
0.01 |
25°C |
3 |
0.25 |
595 |
0.03 |
25°C |
4 |
0.50 |
595 |
0.09 |
25°C |
5 |
1.00 |
595 |
0.21 |
25°C |
6 |
2.00 |
595 |
0.39 | 25°C |
7 |
4.00 |
595 |
0.76 | 25°C |
8 |
6.00 |
595 |
1.20 |
25°C |
9 |
8.00 |
595 |
1.54 |
25°C |
10 |
10.00 |
595 |
1.81 |
25°C |
The absorbance and concentrations were recorded at595 nm.
0 Hours | 0 Hours | 72 Hours | 72 Hours | ||
---|---|---|---|---|---|
SR. No | concentrations (mg/L) | Absorbance (mean) | Analytical concentrations | Absorbance (mean) | Analytical concentrations |
1 | control | 0.00 | 0.00 | 0.00 | 0.00 |
2 | 0.2 | 0.04 | 0.22 | 0.02 | 0.13 |
3 | 0.54 | 0.11 | 0.59 | 0.07 | 0.37 |
4 | 1.46 | 0.29 | 1.54 | 0.23 | 1.23 |
5 | 3.94 | 0.79 | 4.17 | 0.48 | 2.54 |
6 | 10.64 | 1.11* | 11.72 | 0.64* | 6.8 |
Note: Dilutions were made in case of*marked absorbance. Concentration values are recorded accordingly. Average of 0thand 3rdday ANDT for each concentration is given below: |
|||||
Sr. No. | Nominal Concentration (mg/L) | Measured Concentration in mg/L (calculated as average of Analytical determination readings of 0thand 3rdDay) | |||
1 | 0.2 | 0.18 | |||
2 | 0.54 | 0.48 | |||
3 | 1.46 | 1.39 | |||
4 | 3.94 | 3.36 | |||
5 | 10.64 | 9.26 | |||
The test concentrations were measured and found not to remain within 80 – 120 % of nominal. Hence, the ErC 50 was expressed based on measured concentrations.
Test Concentration(mg/L) | Experimental Flasks | pH | Temperature °C | ||
---|---|---|---|---|---|
0 Hours | 72 Hours | 0 Hours | 72 Hours | ||
control | R1 | 7.32 | 7.95 | 25 | 25 |
control | R2 | 7.36 | 7.98 | 25 | 25 |
control | R3 | 7.37 | 7.92 | 25 | 25 |
0.2 | R1 | 7.46 | 7.97 | 25 | 25 |
0.2 | R2 | 7.40 | 7.92 | 25 | 25 |
0.2 | R3 | 7.39 | 7.82 | 25 | 25 |
0.54 | R1 | 7.36 | 7.89 | 25 | 25 |
0.54 | R2 | 7.37 | 7.90 | 25 | 25 |
0.54 | R3 | 7.39 | 7.98 | 25 | 25 |
1.46 | R1 | 7.39 | 7.94 | 25 | 25 |
1.46 | R2 | 7.38 | 7.93 | 25 | 25 |
1.46 | R3 | 7.37 | 7.92 | 25 | 25 |
3.94 | R1 | 7.30 | 7.99 | 25 | 25 |
3.94 | R2 | 7.31 | 7.86 | 25 | 25 |
3.94 | R3 | 7.34 | 7.89 | 25 | 25 |
10.64 | R1 | 7.38 | 7.92 | 25 | 25 |
10.64 | R2 | 7.36 | 7.86 | 25 | 25 |
10.64 | R3 | 7.33 | 7.89 | 25 | 25 |
The pH was measured at the beginning of the test and after 72 hr of exposure. The pH of the control medium did not increase by more than 1.5 units during the test.
Experimental Flasks and Test Concentration(mg/L) |
0 Hr Cell
Count
|
24 Hr
Count
Cell |
48 Hr
Cell Count
|
72 Hr
Cell Count
|
Avg Specific Growth Rate (µ) | Mean Avg Specific Growth Rate (µ) | Percent Inhibition(%) |
---|---|---|---|---|---|---|---|
control | 10000 | 45000 | 180000 | 490000 | 1.30 | 1.30 | - |
control | 10000 | 40000 | 160000 | 475000 | 1.29 | ||
control | 10000 | 40000 | 160000 | 525000 | 1.32 | ||
0.2 | 10000 | 35000 | 85000 | 100000 | 0.77 | 0.76 | 41.90 |
0.2 | 10000 | 40000 | 80000 | 95000 | 0.75 | ||
0.2 | 10000 | 30000 | 80000 | 95000 | 0.75 | ||
0.54 | 10000 | 30000 | 60000 | 65000 | 0.62 | 0.60 | 54.17 |
0.54 | 10000 | 30000 | 50000 | 60000 | 0.60 | ||
0.54 | 10000 | 30000 | 50000 | 55000 | 0.57 | ||
1.46 | 10000 | 30000 | 45000 | 55000 | 0.57 | 0.57 | 56.34 |
1.46 | 10000 | 25000 | 40000 | 55000 | 0.57 | ||
1.46 | 10000 | 25000 | 40000 | 55000 | 0.57 | ||
3.94 | 10000 | 20000 | 35000 | 50000 | 0.54 | 0.49 | 62.59 |
3.94 | 10000 | 20000 | 35000 | 40000 | 0.46 | ||
3.94 | 10000 | 20000 | 30000 | 40000 | 0.46 | ||
10.64 | 10000 | 10000 | 15000 | 20000 | 0.23 | 0.23 | 82.25 |
10.64 | 10000 | 10000 | 15000 | 20000 | 0.23 | ||
10.64 | 10000 | 15000 | 20000 | 20000 | 0.23 |
Description of key information
A freshwater algal growth inhibition test was conducted for 72 hrs for assessing the effect of test chemical on green algae Pseudokirchneriella subcapitata. The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. OECD medium composed of macronutrients, micronutrients, alkaline EDTA solution and iron solution was used as a growth medium. The test solution was prepared by dissolving 1000 mg of test chemical in 1000 ml of OECD media, and allowed for stirring for 96 hours, which was then filtered and the final stock solution obtained was 1000 mg/L, verified analytically by UV-Vis Spectrophotometer. The final solubility value obtained after analytical detection is 13.71 mg/L. To have a better growth and visibility of cells, the initial cell count of the culture was kept 10,000 cells/mL. Further, exposure concentrations of 0, 0.2, 0.54, 1.46, 3.94 and 10.64 mg/L (factor of 2.7), respectively was from the stock test concentrations. Test vessel were placed in orbital shaking incubator for 72 hrs at a temperature of 21 to 24 ± 2°C and with a continuous uniform illumination of 6000 -10000 lux light intensity, respectively. The speed of the orbital shaking incubator was set at a 120 revolutions per minute throughout the study period. The cultures were counted and observed daily with the help of a microscope. Potassium dichromate (K2Cr2O7) was used as a reference substance. The 72 hr EC50 value of the reference substance was determined to be 0.868 mg/l. The biomass in the control vessel have increased exponentially by a factor of 16, the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% (i.e., reported to be 1.31%) and the mean coefficient of variation of specific growth rate was not exceeded 35% (i.e., reported to be 4.44%), respectively thus fulfilling the validity criterion. As the concentration of the test chemical being tested has not been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on measured concentration. On the basis of growth rate of the test organism, the 72 hr median effect concentration (ErC50) value was determined to be 0.52 mg/l (nominal conc.) and 0.47 mg/l (measured conc.), respectively. Thus, test chemical was considered as toxic to aquatic algae at environmental relevant concentrations and hence, considered to be classified in 'aquatic acute/chronic category 1' as per the CLP classification criteria.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 0.47 mg/L
Additional information
A freshwater algal growth inhibition test was conducted for 72 hrs for assessing the effect of test chemical on green algae Pseudokirchneriella subcapitata. The test was performed in accordance to OECD guideline No. 201 – Alga growth inhibition test under static condition. Initial cell density of the culture was kept at 10000 cells/ml. OECD medium composed of macronutrients, micronutrients, alkaline EDTA solution and iron solution was used as a growth medium. The test solution was prepared by dissolving 1000 mg of test chemical in 1000 ml of OECD media, and allowed for stirring for 96 hours, which was then filtered and the final stock solution obtained was 1000 mg/L, verified analytically by UV-Vis Spectrophotometer. The final solubility value obtained after analytical detection is 13.71 mg/L. To have a better growth and visibility of cells, the initial cell count of the culture was kept 10,000 cells/mL. Further, exposure concentrations of 0, 0.2, 0.54, 1.46, 3.94 and 10.64 mg/L (factor of 2.7), respectively was from the stock test concentrations. Test vessel were placed in orbital shaking incubator for 72 hrs at a temperature of 21 to 24 ± 2°C and with a continuous uniform illumination of 6000 -10000 lux light intensity, respectively. The speed of the orbital shaking incubator was set at a 120 revolutions per minute throughout the study period. The cultures were counted and observed daily with the help of a microscope. Potassium dichromate (K2Cr2O7) was used as a reference substance. The 72 hr EC50 value of the reference substance was determined to be 0.868 mg/l. The biomass in the control vessel have increased exponentially by a factor of 16, the coefficient of variation of average specific growth rates during the whole test period in replicate control cultures must not exceed 7% (i.e., reported to be 1.31%) and the mean coefficient of variation of specific growth rate was not exceeded 35% (i.e., reported to be 4.44%), respectively thus fulfilling the validity criterion. As the concentration of the test chemical being tested has not been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on measured concentration. On the basis of growth rate of the test organism, the 72 hr median effect concentration (ErC50) value was determined to be 0.52 mg/l (nominal conc.) and 0.47 mg/l (measured conc.), respectively. Thus, test chemical was considered as toxic to aquatic algae at environmental relevant concentrations and hence, considered to be classified in 'aquatic acute/chronic category 1' as per the CLP classification criteria.
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