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EC number: 204-442-7 | CAS number: 121-00-6
- 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:
- Experimental test result performed according to the guideline.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Principles of method if other than guideline:
- This study was designed to access the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test).
- GLP compliance:
- no
- Analytical monitoring:
- no
- Remarks:
- As chemical is stable
- Vehicle:
- no
- Details on test solutions:
- The test solution was prepared in aseptic condition. The test substance tert-butyl-4-methoxyphenol was prepared by adding 10 mg of test substance in 100 ml of BBM to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104 cells/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:
- 24 °C ±2°C.
- pH:
- 6.7 to 7.2
- Nominal and measured concentrations:
- 1.5, 2.7, 4.86, 8.748, 15.75 and 28.34 mg/L. All the six concentration were in geometric series spaced by a factor of 1.8.
- Details on test conditions:
- Light Quality White Fluoroscent Light
Light Intensity 1500Lux
Photoperiod 16 Hour Light Period : 8 Hour Dark Period
RPM Speed 120 Revolutions per minute
Water Deionized Waer
Test vessel size 100ml
Test solution volume 60ml
Dilution water Bold’s Basal Medium
Test concentration Six test concentration were: 1.5, 2.7, 4.86, 8.748, 15.75 and 28.34 mg/L
Replicates per concentration
Three replicates for Control
Two replicates for each test concentration - Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 9.05 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: calculation from equation
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 9.33 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: calculation from graph
- Details on results:
- The microscopic observations were also noted in each of the experimental flasks. All the cells appeared healthy, round and green throughout the test duration in the control while in the experimental flask following changes were observed with increase in test substance concentration.
Decrease in cell count
Discoloration of algal cells
Reduced cell size
Fine particles observed inside the cell - Validity criteria fulfilled:
- yes
- Conclusions:
- After 72 hours of exposure to test substance to various nominal test concentration, EC50 calculated from equation and graphically through probit analysis was observed to be 9.05 mg/L and 9.33 mg/L respectively.
- Executive summary:
This study was designed to access the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test). The test solution was prepared in aseptic condition. The test substance Tert-butyl-4-methoxyphenol was prepared by adding 10 mg of test substance in 100 ml of BBM to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment. The effect of the test substance on the green algae Chlorella vulgaris culture was observed at nominal test concentration of 1.5, 2.7, 4.86, 8.748, 15.75 and 28.34 mg/L. For the assessment of algal growth, the test was conducted in replicates. The control flask was maintained in triplicates as recommended in the OECD guideline and the test concentration were selected in geometric series which were maintained in duplicates. 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) were determined. The microscopic observations were also noted in each of the experimental flasks. All the cells appeared healthy, round and green throughout the test duration in the control while following changes were observed at the higher concentrations. Decreased in cell count and reduced cell size. After 72 hours of exposure to test substance to various nominal test concentration, EC50 calculated from equation and graphically through probit analysis was observed to be 9.05 mg/L and 9.33 mg/L respectively.
Reference
Table 1: Showing the average cell count using Haemocytometer of the experimental flasks at an equal interval of 24hrs, 48hrs and 72hrs
Experimental Flasks and test concentration |
24 Hours |
48 Hours |
72 Hours |
Control |
|||
Replicate 1 |
49.5 x 104 |
66 x 104 |
10.5 x 105 |
Replicate 2 |
46 x 104 |
59 x 104 |
10.55 x 105 |
Replicate 3 |
54.5 x 104 |
66.5 x 104 |
10.8 x 105 |
Test chemical |
|||
1.5 mg/l |
|||
Replicate 1 |
41.5 x 104 |
50.5 x 104 |
47 x 104 |
Replicate 2 |
44.5 x 104 |
48.5 x 104 |
59.5 x 104 |
2.7 mg/l |
|||
Replicate 1 |
56 x 104 |
43 x 104 |
44 x 104 |
Replicate 2 |
48 x 104 |
35.5 x 104 |
40 x 104 |
4.86 mg/l |
|||
Replicate 1 |
39 x 104 |
40 x 104 |
28 x 104 |
Replicate 2 |
36.5 x 104 |
43.5 x 104 |
30 x 104 |
8.75 mg/l |
|||
Replicate 1 |
32.5 x 104 |
37 x 104 |
9 x 104 |
Replicate 2 |
44 x 104 |
37 x 104 |
10.5 x 104 |
15.75 mg/l |
|||
Replicate 1 |
39.5 x 104 |
35.5 x 104 |
5 x 104 |
Replicate 2 |
44.5 x 104 |
34.5 x 104 |
8.5 x 104 |
28.34 mg/l |
|||
Replicate 1 |
46.5 x 104 |
38.5 x 104 |
3 x 104 |
Replicate 2 |
41 x 104 |
33 x 104 |
2.5 x 104 |
Table 2 : Showing the values of average specific growth rate and percentage inhibition after an interval of 72 hours
|
CONTROL |
1.5 mg/l |
2.7 mg/l |
4.86 mg/l |
8.75 mg/l |
15.75 mg/l |
28.34 mg/l |
||||||||
Average Specific Growth rate (µ ) |
R1 |
1.551 |
R1 |
1.283 |
R1 |
1.261 |
R1 |
1.110 |
R1 |
0.732 |
R1 |
0.536 |
R1 |
0.366 |
|
R2 |
1.552 |
R2 |
1.361 |
R2 |
1.229 |
R2 |
1.33 |
R2 |
0.783 |
R2 |
0.713 |
R2 |
0.305 |
|
|
R3 |
1.560 |
|
|||||||||||||
Mean of Avg. Specific growth rate |
1.554 |
1.322 |
1.245 |
1.122 |
0.758 |
0.624 |
0.335 |
||||||||
Percentage Inhibition (%I) |
_ |
14.938 |
19.901 |
27.829 |
51.246 |
59.811 |
78.403 |
Description of key information
This study was designed to access the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test). The test solution was prepared in aseptic condition. The test substance Tert-butyl-4-methoxyphenol was prepared by adding 10 mg of test substance in 100 ml of BBM to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment. The effect of the test substance on the green algae Chlorella vulgaris culture was observed at nominal test concentration of 1.5, 2.7, 4.86, 8.748, 15.75 and 28.34 mg/L. For the assessment of algal growth, the test was conducted in replicates. The control flask was maintained in triplicates as recommended in the OECD guideline and the test concentration were selected in geometric series which were maintained in duplicates. 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) were determined. The microscopic observations were also noted in each of the experimental flasks. All the cells appeared healthy, round and green throughout the test duration in the control while following changes were observed at the higher concentrations. Decreased in cell count and reduced cell size. After 72 hours of exposure to test substance to various nominal test concentration, EC50 calculated from equation and graphically through probit analysis was observed to be 9.05 mg/L and 9.33 mg/L respectively.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 9.33 mg/L
Additional information
Based on the various data for the test chemical, studies were reviewed for the toxicity evaluation on the growth of algae and cyanobacteria end point which are summarized as below:
This study was designed to access the toxic effects of the test compound on the green alga Chlorella vulgaris. Test was conducted in compliance with the OECD guideline 201 (Alga, Growth Inhibition Test). The test solution was prepared in aseptic condition. The test substance Tert-butyl-4-methoxyphenol was prepared by adding 10 mg of test substance in 100 ml of BBM to get the final concentration of 100 mg/L. The remaining test solutions were prepared by dilution from the above stock solution. To have a better growth and visibility of cells, the initial cell density of the culture was kept 1 X 104cells/ml. Care was taken to have a homogeneous solution for the experiment. The effect of the test substance on the green algae Chlorella vulgaris culture was observed at nominal test concentration of 1.5, 2.7, 4.86, 8.748, 15.75 and 28.34 mg/L. For the assessment of algal growth, the test was conducted in replicates. The control flask was maintained in triplicates as recommended in the OECD guideline and the test concentration were selected in geometric series which were maintained in duplicates. 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) were determined. The microscopic observations were also noted in each of the experimental flasks. All the cells appeared healthy, round and green throughout the test duration in the control while following changes were observed at the higher concentrations. Decreased in cell count and reduced cell size. After 72 hours of exposure to test substance to various nominal test concentration, EC50 calculated from equation and graphically through probit analysis was observed to be 9.05 mg/L and 9.33 mg/L respectively.
Determination of short term toxicity of test chemical on the growth of aquatic algae and cyanobacteria. Test conducted for 72 hrs. Effect were observed on the basis of growth rate inhibition and area under the growth curve. The 72 hour EC50 value of test chemical in green algae was determine to be 1.9 mg/L based on its effect on the growth rate (area under the growth curve) and 5.2 on the basis of growth rate. The NOEC was observed at 0.25 mg/l. Thus based on the EC50 value, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.
Thus based on the above effects, chemical consider to be toxic and classified as aquatic chronic 2 as per the CLP classification criteria.
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