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EC number: 603-094-7 | CAS number: 125904-11-2
- 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
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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
- Study period:
- from November 16, 2004 to November 23, 2004
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Study conducted according to OECD Guideline 201 and mostly in accordance with GLP standards.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 850.5400 (Algal Toxicity, Tiers I and II) (January 2012)
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- GLP compliance:
- yes
- Remarks:
- with the following exceptions: The characterization and the stability of the test substance under conditions of storage at the test site were not determined in compliance with GLP Standards.
- Analytical monitoring:
- yes
- Details on sampling:
- - Sampling method: Samples of the test solutions were collected at approximately 0 and 96 hours to measure the concentrations of the test substance. Samples at test initiation were collected from the individual batches of test solution prepared for each treatment and control group prior to addition of the algae. At exposure termination, samples were collected from the pooled replicated from each treatment and control group.
- Sample storage conditions before analysis: All samples were collected in glass vials and were processed on the day of collection and analysed as soon as possible. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:A primary stock solution was prepared by dissolving DBS in freshwater algal medium at a nominal concentration of 10 mg/L. The stock was inverted at least 20 times, sonicated and stirred on a magnetic stir plate with a Teflon® coated stir bar for approximately 24 hours to mix and appeared clear and colorless. The primary stock was proportionally diluted with freshwater algal medium to prepare the four additional test solutions at nominal concentrations of 0.63, 1.3,2.5 and 5.0 mg/L. Test concentrations were not corrected for percent active ingredient in the test substance.
- Controls: A negative control was performed
- Evidence of undissolved material: All test solutions appeared clear and colorless - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Freshwater green alga, Selenastrum capricornutum
- Strain:Printz (UTCC 37)
- Source (laboratory, culture collection): University of Toronto Culture Collection
- Age of inoculum : cultures had been actively growing in culture medium for at least two weeks prior to test initiation
- Method of cultivation: The algal cells were cultured and tested in freshwater algal medium. The culture was last transferred to fresh medium four days prior to test initiation. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 96 h
- Hardness:
- not measured
- Test temperature:
- Temperatures ranged from 23.8 to 25.2°C and were within the 24 ± 2°C range established for the test. The minimum and maximum temperatures measured continuously throughout the study were 23 and 24°C, respectively.
- pH:
- The pH of the test solutions at test initiation was 8.0 and at exposure termination ranged from 8.4 to 9.4. The pH tended to increase relative to increases in algal densities, which is typical for tests conducted with Selenastrum capricornutum.
- Dissolved oxygen:
- not measured
- Salinity:
- NA
- Nominal and measured concentrations:
- Nominal concentrations selected for use in this study were 0.63, 1.3, 2.5, 5.0 and 10 mg/L. Samples collected on Day 0 had recoveries of 62, 59, 63, 63 and 67% of nominal concentrations, respectively. Recoveries in all samples collected at 96 hours, were
- Details on test conditions:
- TEST SYSTEM
- Test vessel: sterile, 250-mL Erlenmeyer flasks plugged with foam stoppers containing 100 mL of test or control mediu
- Type (delete if not applicable): closed
- Initial cells density: 10000 cells/mL
- Control end cells density: 3,195,748 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3
GROWTH MEDIUM
Stock nutrient solutions were prepared by adding reagent-grade chemicals to purified Wildlife International, Ltd. well water. The test medium then was prepared by adding appropriate volumes of the stock nutrient solutions to purified well water, NANOpure® water, (NH4C1 15 mg/L, MgC12.6H2O 12 mg/L, CaC12.2H2O 18 mg/L, MgSO4.7H2O 15 mg/L, KH2PO4 1.6 mg/L, FeC13.6H2O 0.08 mg/L, Na2EDTA•2H2O 0.1 mg/L, H3BO3 0.185 mg/L, MnC12.4H20 0.415 mg/L ZnC12 3 µg/L, CoC12.6H2O 1.5 µg/L, CuC12.2H2O 0.01 µg/L, Na2MoO4.2H2O 7 µg/L and
NaHCO3 50 mg/L.)
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Wildlife International, Ltd. well water
- Pesticides, organics and metals:
In µg/L: Aldrin < 0.0020, Heptachlor Epoxide < 0.0020, Alpha BHC < 0.0020, Malathion < 0.39, Beta BHC < 0.012, Merphos < 0.39, Bolstar < 0.39, Methoxychlor < 0.020, Chlordane < 0.068, Methyl Parathion < 0.39, Coumaphos < 0.62, Mevinphos < 0.39, Delta BHC < 0.029, Mirex < 0.0098, Demeton-O < 0.39, Naled < 0.58, Demeton-S < 0.39, o,p-DDD < 0.0039, Diazinon < 0.39, o,p-DDE < 0.0039, Dichlorvos < 0.39, o,p-DDT < 0.0039, Dieldrin < 0.0049, p,p-DDD < 0.0039, Disulfoton < 0.39, p,p-DDE < 0.0039, Dursban (Chlorpyrifos) < 0.39, p,p-DDT < 0.0039, Endosulfan I < 0.0039, PCB-1016 < 0.20, Endosulfan II < 0.0049, PCB-1221 < 0.39, Endosulfan Sulfate ,< 0.0088, PCB-1232 < 0.098, Endrin <0.0039, PCB-1242 < 0.20, EPN < 0.78, PCB-1248 < 0.29, Ethion < 0.39, PCB-1254 < 0.20, Ethoprop < 0.39, PCB-1260
< 0.29, Ethyl Parathion < 0.39, Phorate < 0.39, Famphur < 0.48, Ronnel < 0.39, Fensulfothion < 0.87, Stirophos < 0.39, Fenthion < 0.39, Telodrin < 0.0020, Gamma BHC – Lindane < 0.0020, Tokuthion < 0.39, Guthion (Azinphos-methyl) < 0.78, Toxaphene < 0.29, HCB < 0.0020, Trichloronate < 0.39, Heptachlor < 0.0020 and Trithion < 0.39
In mg/L: Aluminum <0.0413, Antimony < 0.0085, Arsenic <0.0049, Barium <0.005, Beryllium <0.00034, Bromide <2.0, Cadmium <0.00087, Calcium 36.2, Chloride 3.4, Chromium < 0.0022, Cobalt <0.0016, Copper <0.0021, Fluoride 0.58, Iron <0.0453, Lead <0.0093, Magnesium 13.7, Manganese <0.005, Mercury <0.00016, Nickel <0.0038, Nitrate Nitrogen <0.40, Potassium 6.20, Selenium <0.0047, Silver < 0.0018, Sodium 19.3, Sulfate 4.7, Thallium <0.0089, Vanadium <0.0017, and Zinc <0.0041
- Culture medium different from test medium:
- Intervals of water quality measurement:
OTHER TEST CONDITIONS
- Sterile test conditions: yes, the medium was sterilized by filtration (0.22 µm) prior to use.
- Adjustment of pH: ). The pH of the medium was adjusted to 7.9 using 0.1N NaOH
- Photoperiod: 24 hours/day
- Light intensity and quality: Cool-white fluorescent lighting at an intensity of 4300 ± 10% lux. Light intensity was measured at five locations surrounding the test flasks on the shaker table at test initiation. Light intensity was measured using a SPER Scientific 840006C light meter. The light intensity ranged from 4100 to 4620 lux, which was within the desired range of 4300 ± 10% lux
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: The concentration of algal cells was verified using a haemacytometer and microscope, and 1.0 mL of the inoculum was added to each test chamber to achieve a nominal concentration of approximately 10,000 cells/mL at test initiation.
TEST CONCENTRATIONS
- Range finding study: A range finding toxicity study was performed, the results were not presented in the reporting of the study.
- Test concentrations: Nominal test concentrations selected were 0.63, 1.3, 2.5, 5.0 and 10 milligrams DBS per liter (mg/L).- Reference substance (positive control):
- not required
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 6.2 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: cell density
- Remarks on result:
- other: 5.4-7.0 mg/L (95% CL)
- Duration:
- 96 h
- Dose descriptor:
- EC50
- Effect conc.:
- 7.3 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: cell density
- Remarks on result:
- other: 6.5-8.2 mg/L (95% CL)
- Duration:
- 72 h
- Dose descriptor:
- other: NOAEC
- Effect conc.:
- 0.63 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: cell density
- Duration:
- 96 h
- Dose descriptor:
- other: NOAEC
- Effect conc.:
- 2.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: cell density
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 6.1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 5.4-6.9 mg/L (95% CL)
- Duration:
- 96 h
- Dose descriptor:
- EC50
- Effect conc.:
- 6.9 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 6.2-7.7 mg/L (95% CL)
- Duration:
- 72 h
- Dose descriptor:
- other: NOAEC
- Effect conc.:
- 0.63 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 96 h
- Dose descriptor:
- other: NOAEC
- Effect conc.:
- 2.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 9.1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 8.5-9.7 mg/L (95% CL)
- Duration:
- 96 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: CL not calculable
- Duration:
- 72 h
- Dose descriptor:
- other: NOAEC
- Effect conc.:
- 2.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 96 h
- Dose descriptor:
- other: NOAEC
- Effect conc.:
- 5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- - Exponential growth in the control (for algal test): YES
- Observation of abnormalities (for algal test): some enlarged cells were noted for the 10 mg/L treatment group
- Any stimulation of growth found in any treatment: NO.
Nominal concentrations selected for use in this study were 0.63,1.3, 2.5, 5.0 and 10 mg/L. Samples collected on Day 0 had recoveries of62, 59,63,63 and 67% of nominal concentrations, respectively. Recoveries in all samples collected at 96 hours, were
- Effect concentrations exceeding solubility of substance in test medium: YES - This is not discussed in the present report; however, the dominant (85%) dibromostyrene constituent has a water solubility of 6.91 mg/L according to the key study on solubility (endpoint summary 4.8). Therefore, some of the reported EC50s for the present study exceed this level. This may be because the ECs are reported in terms of nominal concentrations, while actual measured concentrations in the present study were much less.- Reported statistics and error estimates:
- Please refer to subheading Statistical analyses under section Any other information on materials and methods incl. tables.
Table 1
Mean Cell Density and Percent InhibitionNominal
24 Hours
48 Hours
72 Hours
96 Hours
Test Concentration (mg/L)
MeanCell Density(cells/mL)
Percent Inhibition1,2
Mean Cell Density (cell s/mL
Percent Inhibition1,2
Mean Cell Density (cells/mL)
Percent Inhibition1,2
Mean Cell Density (cells/mL)
Percent Inhibition1,2
Negative Control
17,157
117,044
757,937
3,195,748
0.63
15,737
8.3
103,376
12
700,128
7.6
3,146,431
1.5
1.3
16,675
2.8
100,729
14
631,686
17
2,940,067
8.0
2.5
14,996
13
101,600
13
579,896*
23
2,962,938
7.3
5.0
11,443
33
84,895
27
487,148*
36
2,692,040
16
10
14,046
18
12,862
89
58,219*
92
529,676*
83
1Calculations were performed using SAS Version 8.02. Manual calculations may differ slightly.2Percent inhibition was calculated relative to the negative control replicates.
* Statistically significant difference (p<0.05) at 72 and 96 hours from the negative control replicates using Dunnett's test.
Table 2
Mean Area Under the Growth Curve (Biomass) and Percent InhibitionNominal
0 - 24 Hours
0 - 48 Hours
0 - 72 Hours
0 - 96 Hours
Test Concentration
(mg/L)
Mean
Area1
Percent
Inhibition1,2
Mean
Area1
Percent
inhibition1,2
Mean
Area1
Percent
Inhibition1,2
Mean
Area1
Percent
Inhibition1,2
Negative Control
85,880
1,456,288
11,716,064
58,920,292
0.63
68,848
20
1,258,212
14
10,660,264
9.0
56,578,968
4.0
1.3
80,104
6.7
1,248,956
14
9,797,940
16
52,418,980
11
2.5
59,956
30
1,219,108
16
9,157,056*
22
51,431,068
13
5.0
22,296
`74
938,348
36
7,562,868*
35
45,473,124*
23
10
48,552
43
131,448
91
744,424*
94
7,559,168*
87
1Calculations were performed using SAS Version 8.02. Manual calculations may differ slightly.
2Percent inhibition was calculated relative to the negative control replicates.
* Statistically significant difference (p<0.05) at 72 and 96 hours from the negative replicates using Dunnett's test.
Table3
Mean Growth Rate and Percent InhibitionNominal
0 - 24 Hours 0 - 48 Hours
0 - 72 Hours
0 - 96 Hours
Test Concentration
(mg/L)
Mean
Growth Rate1
Percent
Inhibition1,2
Mean
Growth Rate1
Percent
Inhibition1,2
Mean
Growth Rate1
Percent
Inhibition1,2
Mean
Growth Rate1
Percent
Inhibition1,2
Negative Control
0.0223
0.0512
0.0600
0.0600
0.63
0.0187
16
0.0485
5.3
0.0590
1.7
0.0599
0.11
1.3
0.0213
4.5
0.0480
6.2
0.0575
4.1
0.0592
1.3
2.5
0.0168
25
0.0483
5.8
0.0564
6.0
0.0592
1.3
5.0
0.0068
69
0.0445
13
0.0538*
10
0.0582
2.9
10
0.0140
37
0.0052
90
0.0237*
61
0.0412*
31
1Calculations were performed using SAS Version 8.02. Manual calculations may differ slightly.
2Percent inhibition was calculated relative to the negative control replicates.
* Statistically significant difference (p<0.05) at 72 and 96 hours from the negative control replicates using Dunnett's test.
Table 4
EC50, EbC50 and ErC50 Values Over the 96-Hour Exposure PeriodCell Density
Area Under the Growth Curve
(Biomass)
Growth Rate
Time
EC50
(mg/L)
95% Confidence
Interval
(mg/L)
EbC50
(mg/L)
95% Confidence
Interval
(mg/L)
ErC50
(mg/L)
95% Confidence
Interval
(mg/L)
24 Hours
>10
--1
>10
--1
>10
--1
48 Hours
6.7
6.1 and 7.3
6.3
4.3 and 7.6
7.1
6.7 and 7.6
72 Hours
6.2
5.4 and 7.0
6.1
5.4 and 6.9
9.1
8.5 and 9.7
96 Hours
7.3
6.5 and 8.2
6.9
6.2 and 7.7
>10
--1
' 95% confidence limits could not be calculated with the data obtained.
- Validity criteria fulfilled:
- yes
- Remarks:
- OECD 201: The cell concentration in the control cultures should have increased by a factor of at least 16 within three days; Disappearance of the test substance from the water into the biomass does not necessarily invalidate the test.
- Conclusions:
- Selenastrum capricornutum were exposed to five concentrations of DBS and evaluated for effects on cell density, area under the growth curve (biomass) and growth rate. Biomass was the most sensitive endpoint, as defined by the lowest 96-hour EC50 value. The 72-hour EbC50, based on biomass, was 6.1 mg/L, while the calculated EC50 and ErC50 values were 6.2 and 9.1 mg/L, respectively. The 96-hour EbC50, based on biomass, was 6.9 mg/L, while the calculated EC50 and ErC50 values were 7.3 and > 1 0 mg/L, respectively. The 72-hour NOAEC based on cell density and biomass was 0.63 mg/L. The 96-hour NOAEC, based on cell density and biomass was 2.5 mg/L.
- Executive summary:
In a GLP compliant study conducted in accordance with OECD 201, Selenastrum capricornutum were exposed to five concentrations of DBS and evaluated for effects on cell density, area under the growth curve (biomass) and growth rate. Biomass was the most sensitive endpoint, as defined by the lowest 96-hour EC50 value. The 72-hour EbC50, based on biomass, was 6.1 mg/L, while the calculated EC50 and ErC50 values were 6.2 and 9.1 mg/L, respectively. The 96-hour EbC50, based on biomass, was 6.9 mg/L, while the calculated EC50 and ErC50 values were 7.3 and > 1 0 mg/L, respectively. The 72-hour NOAEC based on cell density and biomass was 0.63 mg/L. The 96-hour NOAEC, based on cell density and biomass was 2.5 mg/L.
Reference
Description of key information
Selenastrum capricornutum were exposed to five concentrations of DBS and evaluated for effects on cell density, area under the growth curve (biomass) and growth rate. Biomass was the most sensitive endpoint, as defined by the lowest 96-hour EC50 value. The 72-hour EbC50, based on biomass, was 6.1 mg/L, while the calculated EC50 (cell density) and ErC50 (growth rates) values were 6.2 and 9.1 mg/L, respectively. The 96-hour EbC50, based on biomass, was 6.9 mg/L, while the calculated EC50 (cell density) and ErC50 (growth rate) values were 7.3 and > 1 0 mg/L, respectively. The 72-hour NOAEC based on cell density and biomass was 0.63 mg/L. The 96-hour NOAEC, based on cell density and biomass was 2.5 mg/L.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 10 mg/L
Additional information
The OECD 201, GLP compliant study by Desjardins D et al (2005) was considered adequate and reliable to fulfil the data requirement as a key study. The study was assigned a reliability score of 1.
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