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EC number: 271-524-7 | CAS number: 68583-95-9 This substance is identified in the Colour Index by Colour Index Constitution Number, C.I. 16035:1.
- 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
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Justification for type of information:
- Data is from experimental study report.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Principles of method if other than guideline:
- Short term toxicity study was carried out according to the OECD guideline.
- GLP compliance:
- no
- Analytical monitoring:
- no
- Details on sampling:
- No data available
- Vehicle:
- not specified
- Details on test solutions:
- The test solution was prepared in aseptic condition. The test chemical was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring for 24 hours and filter it to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial of the culture was kept 1 X 10E4cells/ml. Care was taken to have a homogeneous solution for the experiment.
- Test organisms (species):
- Chlorella vulgaris
- Details on test organisms:
- The fresh water green alga Chlorella vulgaris, was used as the test organism. Sterile, unicellular, liquid cultures of algae. The culture was examined under the microscope to confirm that it was unicellular, healthy and not contaminated. The medium to be used for the growth of algae was Bold’s Basal Medium (BBM). It is a medium composed of macronutrients, micronutrients, alkaline EDTA solution and Iron solution. Stock solution of each of these was prepared separately and then a complete medium was prepared and sterilized. De-ionized water was used to prepare the BBM.
- Test type:
- static
- Water media type:
- freshwater
- Total exposure duration:
- 72 h
- Test temperature:
- 22 °C ±2°C.
- pH:
- 6.7 to 7.4
- Nominal and measured concentrations:
- 6.25mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L, 200mg/L. All the six concentration were in geometric series spaced by a factor of 2.
- Details on test conditions:
- TEST SYSTEM
- Test vessel: Conical flask
- Type (delete if not applicable): open / closed: No data
- Material, size, headspace, fill volume: 100 ml of conical flasks filled with 60ml of test solution
- Aeration: no data
- Initial cells density: 38.787 x104 cells/mL)
- Control end cells density: No data
- No. of organisms per vessel: No data
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 3
- No. of vessels per vehicle control (replicates): 3
GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used: No data
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: Yes
- Photoperiod: 16 Hour Light Period : 8 Hour Dark Period
- Light intensity and quality: White Fluorescent Light, 1500Lux
- Salinity (for marine algae): No data
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: spectrophotometer
TEST CONCENTRATIONS
- Spacing factor for test concentrations: No data
- Justification for using less concentrations than requested by guideline: No data
- Range finding study: No data
- Test concentrations: 6.25mg/l,12.5mg/l,25mg/l,50mg/l,100mg/l,200mg/l
- Results used to determine the conditions for the definitive study: Cell growth inhibition - Reference substance (positive control):
- not specified
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 136.57 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: calculated from equation through probit analysis
- Details on results:
- The microscopic observations were noted down in each of the control vessel. All the cells appeared healthy, round and green throughout the study duration in the control. Also, the drift in pH in the control vessels did not increase by >1.5 units when observed on 72 hours as compared to 0 hours. The average pH drift observed in the control vessels was 0.1 units.
- 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
- Conclusions:
- After 72 hours of exposure to test chemical to various nominal test concentration, EC50 calculated from equation through probit analysis was determine to be 136.57 mg/L.
- Executive summary:
The effect of test chemical was studied on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga, growth inhibition test. The test concentration chosen for the study were 6.25mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L, 200mg/L. The test solution was prepared in aseptic condition. The test chemical was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring for 24 hours and filter it to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial of the culture was kept 1 X 104cells/ml.
The microscopic observations were noted down in each of the control vessel. All the cells appeared healthy, round and green throughout the study duration in the control. Also, the drift in pH in the control vessels did not increase by >1.5 units when observed on 72 hours as compared to 0 hours. The average pH drift observed in the control vessels was 0.1 units.The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of the test item. EC50 calculated graphically through probit analysis was observed to be 136.57 mg/L.
Based on the EC50, it can be concluded that the chemical was nontoxic and can be consider to be not classified as toxic as per the CLP classification criteria.
Reference
Table 1: Showing the average cell count using Haemocytometer of the test vessels at an equal interval of 24hrs, 48hrs and 72hrs
Test vessels and test concentration |
24 Hours |
48 Hours |
72 Hours |
Control |
|||
Replicate 1 |
3.72 × 104 |
5.32 × 104 |
6.92 × 104 |
Replicate 2 |
4.08 × 104 |
5.68 × 104 |
7.28 × 104 |
Replicate 3 |
3.96 × 104 |
5.56 × 104 |
7.96 × 104 |
CAS No. 68653 -95 -9 |
|||
6.25mg/l |
|||
Replicate 1 |
27600 |
30000 |
34400 |
Replicate 2 |
29200 |
27600 |
34000 |
12.5mg/l |
|||
Replicate 1 |
26800 |
26800 |
33200 |
Replicate 2 |
25200 |
26000 |
31600 |
25mg/l |
|||
Replicate 1 |
24400 |
25200 |
30800 |
Replicate 2 |
23600 |
25200 |
30000 |
50mg/l |
|||
Replicate 1 |
24400 |
25200 |
30000 |
Replicate 2 |
23200 |
24400 |
30000 |
100mg/l |
|||
Replicate 1 |
22800 |
22800 |
27600 |
Replicate 2 |
22000 |
21200 |
28000 |
200mg/l |
|||
Replicate 1 |
19600 |
20800 |
27200 |
Replicate 2 |
18800 |
19600 |
25200 |
Table 2: Showing the values of average specific growth rate and percentage inhibition after an interval of 72 hours
|
CONTROL |
6.25mg/l |
12.5mg/l |
25mg/l |
50mg/l |
100mg/l |
200mg/l |
||||||||
Average Specific Growth rate (µ ) |
R1 |
0.645 |
R1 |
0.412 |
R1 |
0.400 |
R1 |
0.375 |
R1 |
0.366 |
R1 |
0.338 |
R1 |
0.334 |
|
R2 |
0.662 |
R2 |
0.408 |
R2 |
0.384 |
R2 |
0.366 |
R2 |
0.366 |
R2 |
0.343 |
R2 |
0.308 |
|
|
R3 |
0.691 |
|
|||||||||||||
Mean of Avg. Specific growth rate |
0.666 |
0.410 |
0.392 |
0.371 |
0.366 |
0.341 |
0.321 |
||||||||
Percentage Inhibition (%I) |
_ |
38.457 |
41.178 |
44.356 |
45.014 |
48.827 |
51.829 |
Table 3: Depicting pH values at 0 Hours and after 72 Hours of test item exposure to algae
Test vessels and test concentration |
0 Hours |
72 Hours |
CONTROL |
||
Replicate 1 |
6.15 |
6.30 |
Replicate 2 |
6.25 |
6.34 |
Replicate 3 |
6.35 |
6.40 |
Average |
6.25 |
6.35 |
CAS No. 68583 -95 -9 |
||
6.25mg/l |
||
Replicate 1 |
6.35 |
6.31 |
Replicate 2 |
6.40 |
6.29 |
12.5mg/l |
||
Replicate 1 |
6.50 |
6.50 |
Replicate 2 |
6.45 |
6.52 |
25mg/l |
||
Replicate 1 |
6.49 |
6.53 |
Replicate 2 |
6.47 |
6.52 |
50mg/l |
||
Replicate 1 |
6.48 |
6.55 |
Replicate 2 |
6.50 |
6.53 |
100mg/l |
||
Replicate 1 |
6.42 |
6.50 |
Replicate 2 |
6.45 |
6.52 |
200mg/l |
||
Replicate 1 |
6.49 |
6.56 |
Replicate 2 |
6.50 |
6.57 |
Description of key information
After 72 hours of exposure to test chemical to various nominal test concentration, EC50 calculated from equation through probit analysis was determine to be 136.57 mg/L.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 136.57 mg/L
Additional information
The effect of test chemical was studied on the growth of fresh water green alga Chlorella vulgaris. The study was conducted following OECD guideline 201- Alga, growth inhibition test. The test concentration chosen for the study were 6.25mg/L, 12.5mg/L, 25mg/L, 50mg/L, 100mg/L, 200mg/L. The test solution was prepared in aseptic condition. The test chemical was prepared by adding 50 mg of test item in 250 ml of BBM to get the final concentration of 200 mg/L. This stock solution was kept for stirring for 24 hours and filter it to obtain a homogenous solution for the experiment. The test concentrations were chosen according to the available data of the test item. The concentrations chosen were set up to the water solubility limit. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial of the culture was kept 1 X 104cells/ml.
The microscopic observations were noted down in each of the control vessel. All the cells appeared healthy, round and green throughout the study duration in the control. Also, the drift in pH in the control vessels did not increase by >1.5 units when observed on 72 hours as compared to 0 hours. The average pH drift observed in the control vessels was 0.1 units.The green alga was exposed to the test concentration for a period of 72 hours to observe average specific growth rate and % growth inhibition under the effect of the test item. EC50 calculated graphically through probit analysis was observed to be 136.57 mg/L.
Based on the EC50, it can be concluded that the chemical was nontoxic and can be consider to be not classified as toxic as per the CLP classification criteria.
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