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EC number: 224-924-0 | CAS number: 4553-89-3
- 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 using standard OECD test guidelines
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- GLP compliance:
- no
- Analytical monitoring:
- no
- Vehicle:
- yes
- Details on test solutions:
- The test solution was prepared in aseptic condition. The test item disodium 4,4’-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate was prepared by adding 140 mg of test item in 350 ml of BBM to get the final concentration of 400 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 10(4)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
- Hardness:
- No data
- Test temperature:
- 24 °C ±2°C.
- pH:
- 6.7 to 7.4
- Dissolved oxygen:
- No data
- Salinity:
- No data
- Conductivity:
- No data
- Nominal and measured concentrations:
- 12.5 mg/l, 25 mg/l, 50 mg/l, 100 mg/l, 200 mg/l and 400mg/l. All the six concentration were in geometric series spaced by a factor of 2.
- 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:
Replicates per concentration
Three replicates for Control
Two replicates for each test concentration - Reference substance (positive control):
- not specified
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 379.668 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: calculated graphically through probit analysis.
- Details on results:
- The microscopic observations were also noted in each of the test vessel. All the cells appeared healthy, round and green throughout the study duration and no significant changes were observed up to the concentration of 50 mg/l whereas, black dotted rings were observed inside the algal cells in concentrations of 100mg/l, 200mg/l and 400mg/l. EC50 was found to be 379.668 mg/l graphically through probit analysis
- Validity criteria fulfilled:
- yes
- Conclusions:
- After 72 hours of exposure to various nominal test concentrations of disodium 4,4’-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate (CAS No. 4553-89-3) to freshwater green alga Chlorella vulgaris, EC50 was determined to be 379.668 mg/l graphically through probit analysis.
- Executive summary:
The effect of test item disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate, CAS No. 4553-89-3 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 concentrations chosen for the study were 12.5mg/l, 25mg/l, 50 mg/l, 100mg/l, 200 mg/l and 400mg/l. The test concentrations were prepared using stock solution of the test item using mineral media. 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 379.668 mg/l.
Based on the above LC50 value it can be concluded that the target substance 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate is non toxic to green algae and thus can not be classified under Aquatic toxic category.
Reference
Table 1: Showing the average cell count using Haemocytometer of the experimental flasks at an equal interval of 24hrs, 48hrs and 72hrs
Test vessels and test concentration |
24 Hours |
48 Hours |
72 Hours |
Control |
|||
Replicate 1 |
41.5 x 104 |
70 x 104 |
89 x 104 |
Replicate 2 |
49 x 104 |
65 x 104 |
86 x 104 |
Replicate 3 |
44.5 x 104 |
71.5 x 104 |
91.5x 104 |
CAS No. 4553-89-3 |
|||
12.5mg/l |
|||
Replicate 1 |
36.5 x 104 |
43.5 x 104 |
55.5 x 104 |
Replicate 2 |
41.5 x 104 |
46 x 104 |
58.5 x 104 |
25mg/l |
|||
Replicate 1 |
35 x 104 |
51.5 x 104 |
32 x 104 |
Replicate 2 |
34 x 104 |
35.5 x 104 |
35 x 104 |
50mg/l |
|||
Replicate 1 |
39 x 104 |
43.5x 104 |
29.5 x 104 |
Replicate 2 |
35.5 x 104 |
41 x 104 |
30 x 104 |
100mg/l |
|||
Replicate 1 |
41 x 104 |
43.5 x 104 |
28.5 x 104 |
Replicate 2 |
42.5 x 104 |
40 x 104 |
22 x 104 |
200mg/l |
|||
Replicate 1 |
26 x 104 |
20.5 x 104 |
16.5 x 104 |
Replicate 2 |
27 x 104 |
23 x 104 |
14.5 x 104 |
400mg/l |
|||
Replicate 1 |
16 x 104 |
12 x 104 |
7 x 104 |
Replicate 2 |
12 x 104 |
9.5 x 104 |
9 x 104 |
Table 2: Showing the values of average specific growth rate and percentage inhibition after an interval of 72 hours
|
CONTROL |
12.5mg/l |
25mg/l |
50mg/l |
100mg/l |
200mg/l |
400mg/l |
||||||||
Average Specific Growth rate (µ ) |
R1 |
1.496 |
R1 |
1.339 |
R1 |
1.155 |
R1 |
1.128 |
R1 |
1.117 |
R1 |
0.934 |
R1 |
0.649 |
|
R2 |
1.485 |
R2 |
1.356 |
R2 |
1.185 |
R2 |
1.134 |
R2 |
1.030 |
R2 |
0.891 |
R2 |
0.732 |
|
|
R3 |
1.505 |
|
|||||||||||||
Mean of Avg. Specific growth rate |
1.495 |
1.348 |
1.170 |
1.131 |
1.073 |
0.913 |
0.691
|
||||||||
Percentage Inhibition (%I) |
_ |
9.891 |
21.752 |
24.377 |
28.218 |
38.955 |
53.826 |
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 |
6.5 |
7.1 |
Replicate 2 |
6.6 |
7.0 |
Replicate 3 |
6.6 |
7.0 |
CAS No. 4553-89-3 |
||
12.5 mg/l |
||
Replicate 1 |
6.5 |
6.9 |
Replicate 2 |
6.5 |
6.8 |
25 mg/l |
||
Replicate 1 |
6.5 |
6.8 |
Replicate 2 |
6.5 |
6.7 |
50 mg/l |
||
Replicate 1 |
6.5 |
6.7 |
Replicate 2 |
6.5 |
6.7 |
100 mg/l |
||
Replicate 1 |
6.5 |
6.7 |
Replicate 2 |
6.5 |
6.6 |
200 mg/l |
||
Replicate 1 |
6.5 |
6.6 |
Replicate 2 |
6.5 |
6.6 |
400 mg/l |
||
Replicate 1 |
6.5 |
6.6 |
Replicate 2 |
6.5 |
6.6 |
Description of key information
After 72 hours of exposure to various nominal test concentrations of disodium 4,4’-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate (CAS No. 4553-89-3) to freshwater green alga Chlorella vulgaris, EC50 was determined to be 379.668 mg/l graphically through probit analysis.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 379.668 mg/L
Additional information
Available Experimental and predicted data of the target substance Disodium 4,4’-(2,4-dihydroxy-5- hydroxymethyl-1,3-phenylene bis-azo) di- (naphthalene-1-sulfonate) for the Short term toxicity to aquatic invertebrate are summarized as follows:
For key study, the effect of test item disodium 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate, CAS No. 4553-89-3 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 concentrations chosen for the study were 12.5mg/l, 25mg/l, 50 mg/l, 100mg/l, 200 mg/l and 400mg/l. The test concentrations were prepared using stock solution of the test item using mineral media. 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 379.668 mg/l.
Supporting above experimental study, toxicity to aquatic algae and cyanobacteria of Disodium 4,4’-(2,4-dihydroxy-5- hydroxymethyl-1,3-phenylene bis-azo) di- (naphthalene-1-sulfonate) was estimated using QSAR Toolbox version 3.3. The Effective concentration (LC50) of the substance Disodium 4,4’-(2,4-dihydroxy-5- hydroxymethyl-1,3-phenylene bis-azo) di- (naphthalene-1-sulfonate) in algae Pseudokirchneriella subcapitata in a 72 hour study is estimated to be 155.21 mg/L. based on the effects on population.
Based on the above LC50 values it can be concluded that the target substance 4,4'-[[2,4-dihydroxy-5-(hydroxymethyl)-1,3-phenylene]bis(azo)]bisnaphthalene-1-sulphonate is not likely to be toxic to algae and thus cannot be classified under Aquatic toxic category as per CLP criteria.
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