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EC number: 458-680-3 | CAS number: 797751-44-1 WASOX-VMAC2
- 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:
- February 15 - March 3, 2005
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- According to EU Method C.3, with GLP.
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- GLP compliance:
- yes
- Analytical monitoring:
- no
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
The test substance was added drop by drop to the nutrient medium and was dissolved by stirring for about one hour, followed by filtration of the solution (cellulose acetate filter, 0.2 µm, Sartorius). The solution was filtered because in a previous experiment without filtering, "particles" were formed during the incubation period, interfering with the counting of the algae.
90 mL of the filtrate were combined with 10 mL of the algal inoculum to give the highest test concentration. Aliquots of the filtrate were further diluted with nutrient medium to obtain lower concentrations.
The preparations were made freshly before the start of the exposure.
The test was performed without adjustment of the pH of the test substance cultures.
- Controls:
One negative control culture (reference substance: nutrient medium) was set up concurrently. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: green algae (Selenastrum capricornutum)
- Strain: Selenastrum capricornutum ATCC (American Type Culture Collection) 22662
- Source (laboratory, culture collection): obtained directly from ATCC, 12301 Parklawn Drive, Rockville, Maryland 20852, USA.
- Method of cultivation: Stock cultures, initially cultivated from deep frozen algae, are continuously maintained in 250 mL conical flasks (Erlenmeyer), each containing 100 mL nutrient medium, at a temperature of 23 ± 2°C under permanent light with an intensity of at least 6000 lux. In about weekly intervals 1 mL of the stock culture is diluted 100-fold with nutrient medium for precultivation and incubation is continued.
Precultures were prepared by incubating a new stock culture for three days. At the start of the experiment the cell density was determined and adjusted to about 105 algae/mL by diluting with nutrient medium.
ACCLIMATION
- Culturing media and conditions (same as test or not):
Concentrated nutrient medium:
Stock solution 1 100 mL
Stock solution 2 10 mL
Stock solution 3 10 mL
Stock solution 4 10 mL
sterile deionised water ad 1000 mL
Storage: Refrigerator.
Nutrient medium:
As nutrient medium for the stock cultures, for precultivation and for the test media a tenfold dilution of the "concentrated nutrient medium" with sterile deionised water was used. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Test temperature:
- All the culture and the blank were incubated at 22 ± 1 ºC
- pH:
- The pH was between 7.4 and 7.6 at the start of the incubation in the test cultures and it was 7.2 in the control cultures.
After 72 hours of incubation the pH was 8.3 in the test cultures and it was 8.4 in the control cultures.
The pH in the control cultures changed by 1.2 units during the 72 hours of incubation. - Nominal and measured concentrations:
- Nominal concentrations: 100.0, 50.0, 25.0, 12.5, and 6.3 mg test substance/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 250 mL conical flasks (erlenmeyer)
- Initial cells density: 10 exp.5 algae/mL
- Control end cells density: At incubation time of 72 hours aliquots were taken from each flask (i.e. from each replicate) and the cell densities of the algae were determined by a Coulter Counter
- No. of organisms per vessel: 10 exp4 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
GROWTH MEDIUM
Nutrient medium: Sterile deionised water and sterile concentrated nutrient medium 9+1 (v/v).
Concentrated nutrient medium:
Stock solution 1 100 mL
Stock solution 2 10 mL
Stock solution 3 10 mL
Stock solution 4 10 mL
sterile deionised water ad 1000 mL
OTHER TEST CONDITIONS
- Photoperiod: permanent light
- Light intensity and quality: at least 6000 lux (wavelength 400 - 700 nm)
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: cell counts by a Coulter Counter
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Range finding study
- Test concentrations: As no data were available on algal toxicity, the highest test substance concentration in this study was 100 mg/L, the same as to be used for a limit test. The next four concentrations were each about one half of the previous one. - Reference substance (positive control):
- no
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- dissolved
- Basis for effect:
- cell number
- Remarks on result:
- other: Based on the area under the growth curve
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- dissolved
- Basis for effect:
- growth rate
- Remarks on result:
- other: Based on the average growth rates
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- cell number
- Remarks on result:
- other: Based on the area under the growth curve
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: Based on the average growth rates
- Details on results:
- - Exponential growth in the control (for algal test): The increase in cell density in the control cultures during the 72 hours incubation period was about 82-fold, corresponding to about 6.4 generations and showing suitable growth conditions for the algae.
- Reported statistics and error estimates:
- Based on the areas under the growth curves as well as on the average growth rates two "lowest observed effective concentrations" (LOECs) are calculated by comparison of the data of the three replicates of each test substance culture with the negative control (analysis of variance, followed by the Scheffé-test, P = 0.05). The "no observed effect concentrations" (NOECs) are then derived from these results (highest concentration with no statistically significant difference to the control).
- Validity criteria fulfilled:
- yes
- Conclusions:
- Two (identical) NOEC (0-72h)-values and two (identical) EC50(0-72h)-values were derived:
The NOEC (0-72h) based on the area under the growth curves was 100 mg/mL and the NOEC (0-72h) based on the average growth rates was 100 mg/mL.
The EbC50 (0-72 h) based on the areas under the growth curves was > 100 mg/mL and the ErC50 (0-72 h), based on the average growth rates, was > 100 mg/mL
The concentrations are nominal test substance concentrations. - Executive summary:
An Alga Growth Inhibition test was performed to determine the possible effects of the test substance on the growth of an unicellular green algae, Selenastrum capricomutum (According to EU Method C). Five concentrations (100.0, 50.0, 25.0, 12.5, 6.3 mg of test substance per liter of nutrient medium) were tested. A negative control (nutrient medium only) was included in the test. There were 3 replicates for each test and control culture.
In each vessel the gel density of the algae was determined 24, 48 and 78 hours after the onset of incubation. The pH was measured at the beginning and at the end of the incubation period. During the 72 hours of incubation, the cell density in the control cultures was increased by a factor of about 82 and the pH changed by 1.2 units (from 7.2 to 8.3). No chemical analysis of the actual test substance concentration was performed, neither at the start nor at the end of the exposure.
Two identical NOEC (0-72h) values and two identical EC50(0-72h) values were derived:
The NOEC (0-72h) based on the area under the growth curves was 100 mg/mL and the NOEC (0-72h) based on the average growth rates was 100 mg/mL.
The EbC50(0-72 h) based on the areas under the growth curves was > 100 mg/ml and the ErC50(0-72 h), based on the average growth rates, was > 100 mg/mL .
Reference
Mean areas under the growth curves, average specific growth rates and percent inhibition.
Negative values indicate enhanced growth compared to the control cultures.
concentration of test substance (mg/L) |
mean area under growth curve |
average specific growth rate (µ x 10-3) 0-72 h |
percent inhibition |
|
area |
growth rates |
|||
0 (control) |
14411 |
60.4 |
0.0 |
0.0 |
6.3 |
13800 |
60.0 |
4.2 |
0.7 |
12.5 |
15635 |
61.8 |
-8.5 |
-2.3 |
25.0 |
17031 |
63.0 |
-18.2 |
-4.2 |
50.0 |
15422 |
60.6 |
-7.0 |
-0.4 |
100.0 |
16654 |
61.6 |
-15.6 |
-1.9 |
µ: Specificgrowth rate
Cell densities and mean areas under the growth curves.
Concentration of test substance (mg/L) |
cell density |
Mean cell density (cells x 103/ml |
Mean area under growth curve |
||
(cells x 103/mL |
|||||
replicate 1 |
replicate 2 |
replicate 3 |
|||
Incubation time: 24 h |
0-24 h |
||||
0 (control) |
46 |
58 |
45 |
50 |
480 |
6.3 |
47 |
46 |
49 |
47 |
450 |
12.5 |
52 |
51 |
43 |
49 |
464 |
25.0 |
52 |
46 |
53 |
50 |
485 |
50.0 |
55 |
48 |
35 |
46 |
430 |
100.0 |
63 |
47 |
50 |
53 |
521 |
Incubation time: 48 h |
24-48 h |
||||
0 (control) |
139 |
218 |
146 |
168 |
2373 |
6.3 |
177 |
170 |
182 |
177 |
2448 |
12.5 |
201 |
208 |
146 |
185 |
2564 |
25.0 |
241 |
156 |
229 |
209 |
2868 |
50.0 |
259 |
212 |
118 |
196 |
2665 |
100.0 |
320 |
183 |
207 |
237 |
3241 |
Incubation time: 72 h |
48-72 h |
||||
0 (control) |
619 |
1204 |
623 |
815 |
11558 |
6.3 |
730 |
797 |
729 |
752 |
10902 |
12.5 |
1042 |
1027 |
588 |
886 |
12608 |
25.0 |
982 |
704 |
1168 |
951 |
13678 |
50.0 |
1093 |
1027 |
433 |
851 |
12327 |
100.0 |
1106 |
719 |
749 |
858 |
12892 |
Description of key information
An Alga Growth Inhibition test was performed to determine the possible effects of the test substance on the growth of an unicellular green algae, Selenastrum capricomutum (According to EU Method C). The NOEC (0-72h) based on the area under the growth curves was 100 mg/mL and the NOEC (0-72h) based on the average growth rates was 100 mg/mL.
Key value for chemical safety assessment
- EC10 or NOEC for freshwater algae:
- 100 mg/L
Additional information
Five concentrations (100.0, 50.0, 25.0, 12.5, 6.3 mg of test substance per liter of nutrient medium) were tested. A negative control (nutrient medium only) was included in the test. There were 3 replicates for each test and control culture.
In each vessel the gel density of the algae was determined 24, 48 and 78 hours after the onset of incubation. The pH was measured at the beginning and at the end of the incubation period. During the 72 hours of incubation, the cell density in the control cultures was increased by a factor of about 82 and the pH changed by 1.2 units (from 7.2 to 8.3). No chemical analysis of the actual test substance concentration was performed, neither at the start nor at the end of the exposure.
Two identical NOEC (0-72h) values and two identical EC50(0-72h) values were derived:
The NOEC (0-72h) based on the area under the growth curves was 100 mg/mL and the NOEC (0-72h) based on the average growth rates was 100 mg/mL.
The EbC50(0-72 h) based on the areas under the growth curves was > 100 mg/mland theErC50(0-72 h), based on the average growth rates, was > 100 mg/mL.
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