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EC number: 233-069-2 | CAS number: 10028-15-6
- 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
![](https://echa.europa.eu./o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_ecotoxicological-information.png)
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:
- supporting study
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- unsuitable test system
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- The unicellular green alga Dunaliella tertiolecta was used as model organism to study the efficacy of ozone as potential ballast water treatment . Individual ozone treatment consisted of exposing algal cells to ozone over 5, 10, 15 and 20 min exposure time periods, after which algal cell viability was determined first counting the number of remaining cells using a hemocytometer followed by vital staining techniques to evaluate the cell viability of the algal cells.
- GLP compliance:
- no
- Analytical monitoring:
- no
- Vehicle:
- no
- Test organisms (species):
- Dunaliella tertiolecta
- Details on test organisms:
- D. tertiolecta (Strain # CCMP1320), was cultured to stationary phase in gently aerated artificial seawater (ASW) enriched with Guillard’s f/2 mediumin 2000 ml glass flasks.
- Test type:
- static
- Remarks on exposure duration:
- 5, 10, 15 and 20 min exposure time periods.
- Post exposure observation period:
- No
- Hardness:
- no data
- Test temperature:
- no data
- pH:
- no data
- Dissolved oxygen:
- no data
- Salinity:
- no data
- Conductivity:
- no data
- Nominal and measured concentrations:
- ozone concentrations in each experimental bottle were maintained at 100 ppm
- Details on test conditions:
- 2 L polypropylene vessels; 3 vessels/treatment (experimental and control)/exposure period.
Exposure times 5, 10, 15 and 20 min. Exposure concentration 100 ppm.
During ozone treatment, the experimental or control cultures were stirred using a magnetic stirrer (70 rpm).
Triplicate controls consisted of culturing D. tertiolecta under identical culture conditions and over a similar time period to cultures used in experimental treatment. These cultures were then transferred to 2-l polypropylene bottles for a time period identical to a given experimental exposure period but not exposed to ozone. - Reference substance (positive control):
- no
- Duration:
- 5 min
- Dose descriptor:
- LOEC
- Effect conc.:
- 100 other: ppm
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: mortality
- Details on results:
- Mortality
5 - 10 min: 4%
10 - 20 min: up to 40%
All individual factor experimental treatments caused significantly higher (P < 0.05) levels of mortality in algae than control treatments at all exposure time periods tested. The results are also preseented in figure 1, see below. - Results with reference substance (positive control):
- n/a
- Reported statistics and error estimates:
- Statistical differences in mean percent mortality levels of algal cells in experimental and control treatments for all exposure time periods were determined using an analysis of variance (ANOVA) followed, where appropriate, by Tukey’s HSD post hoc comparison tests.
- Validity criteria fulfilled:
- not applicable
- Remarks:
- study was not performed according to a guideline.
- Conclusions:
- LOEC (5 min) ≤ 100 ppm
- Executive summary:
The unicellular green alga Dunaliella tertiolecta were among other experiments used to study the efficacy of sonication as well as the advanced oxidants, hydrogen peroxide and ozone, as potential ballast water treatments. Combined rather than individual treatments consistently yielded the highest levels of mortality in algal cells (100% over a 2 min exposure) and in brine shrimp (100% and 95% for larvae and adults, respectively, over a 2 min exposure). Single ozone exposure to the alga Dunaliella tertiolecta lead to a mortality of 4% after 5 minutes, which differed significantly from the control. Therefore, it can be concluded that a LOEC is probably below or equals 100 ppm.
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- unsuitable test system
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Cultures of 3 marine algae species were treated with ozone for a short period. Subsequently the growth of the algae was measured during a 5 day period whereafter the algae were reinoculated in untreated water. Hereafter the growth was again measured during a period of 4 or 6 day in two of the three species.
- GLP compliance:
- no
- Analytical monitoring:
- yes
- Details on sampling:
- The sample size varied from 25 to 40 mL, depending upon the exposure time and consequently the total residual oxidants (TRO) levels.
- Vehicle:
- no
- Details on test solutions:
- Prior to initiating the experiments, ozone flow rates were evaluated in test flasks, which contained only filtered seawater, and the rate was adjusted to provide a maximum TRO value of approximately 0.90 mg/L.
- Test organisms (species):
- other: see details on test organisms
- Details on test organisms:
- Nannochloropsis oculata, Isochrysis galbana and Chaetoceros gracilis. All organisms were grown in 50 L cilinder of filtered and autoclaved seawater to produce a concentration of 0.79 x 10E6 cells/mL (Nannochloropsis oculata), 0. 32 x 10E6 cells/mL (Isochrysis galbana) or 0. 31 x 10E6 cells/mL (Chaetoceros gracilis).
- Test type:
- flow-through
- Water media type:
- saltwater
- Limit test:
- no
- Post exposure observation period:
- The cultures were grown for 5 d after treatment.
- Hardness:
- no data
- Test temperature:
- 25 °C
- pH:
- no data
- Dissolved oxygen:
- no data
- Salinity:
- no data
- Nominal and measured concentrations:
- N. oculata
treatment measured TRO
duration (min) (mg/L)
0 0
0.5 0.006
0.75 0.006
1.0 0.014
1.5 0.202
2.0 0.481
I. galbana
treatment measured TRO
duration (min) (mg/L)
0 0
0.5 0
1.0 0.03
1.5 0.08
2.0 0.21
3.0 0.48
C. gracilis.
treatment measured TRO
duration (min) (mg/L)
0 0
0.5 0.01
1.0 0.03
1.5 0.05
2.0 0.10
3.0 0.25 - Details on test conditions:
- - Six treatments, each having a different period of ozone exposure, were utilized for each experiment.
- Four replicates per treatment were used.
- Algal populations were exposed to ozone at the beginning of each experiment. Ozone exposure was carried out in 2-L Pyrex® flasks, which contained a known concentration of algae in approximately 1.8 L of pre-filtered and autoclaved seawater. A small portion of ozone was diverted via norprene tubing from a working ozone generator to the flasks. An adjustable acrylic air flow meter was used to regulate ozone flow, and a fine pore air stone (2.5 X 1.5 cm) was used to diffuse the ozone into the flasks
- Twenty-four 2-L flasks were filled from the stock culture, each with approximately 1.8 L of algae. After treatment the cultures were grown for 5 d with a 24-h photoperiod using flourescent light banks designed to provide approximately 4000 lux. Light aeration was provided for each flask and the temperature was maintained at 25 degrees centigrade.
- A 5 mL sample was taken daily fromeach flask to determine algal concentrations. Cell counts were determined by measuring the absorbance of each sample at a wavelength of 440 nm (N. oculata) or 750 nm (I. galbana and C. gracilis).
- Initial cell densities were 0. 79 x 10E6 cells/mL (Nannochloropsis oculata), 0. 32 x 10E6 cells/mL (Isochrysis galbana) or 0. 31 x 10E6 cells/mL (Chaetoceros gracilis). Four flask were used for each treatment period. - Reference substance (positive control):
- no
- Duration:
- 0.5 min
- Dose descriptor:
- LOEC
- Effect conc.:
- <= 0.06 mg/L
- Nominal / measured:
- meas. (initial)
- Conc. based on:
- other: TRO
- Basis for effect:
- cell number
- Remarks on result:
- other: measured one day after exposure
- Details on results:
- The effects of ozone treatment on three species of phytoplankton were determined by measuring the cell counts of the algae each day for 4-5 d following the day of exposure.
Cell counts for all treatments of N. oculata were significantly lower than that of the control for the first 2 d (Table 1). On d-3, the 0.5 min treatment, which consisted of cultures treated with 0.05 mg/L TRO, was not significantly different from the control. By d-4, those treated at 0.12 mg/L TRO, were not significantly lower in cell count than the control. Additionally, algae treated with 0.24 mg/L TRO had similar cell counts as the control on d-5.
Growth of ozone treated I. galbana was measured daily for 4 d (Table 2). On d-1, only the treatment which produced a TRO level of 0.05 mg/L was not significantly different from the control. Cultures that had TRO levels of 0.23 mg/L showed no significant difference in cell count on d-2, and by d-4, cultures treated with TRO at 0.34 mg/L were similar in cell count to the control, as well.
The growth response of C. gracilis is presented in Table 3. Cell counts for all the treatments were significantly lower than the control through d-2. On d-3, only the 0.5 min treatment (TRO = 0.06 mg/L) was not significantly lower. On d-4, the treatment at an initial TRO of 0.21 mg/L also was not significantly different from the control, and by d-5, the 1.5 min treatment (TRO = 0.31 mg/L) had cell counts similar to the control. - Results with reference substance (positive control):
- n/a
- Reported statistics and error estimates:
- Two-way analysis of variance was used to determine significant differences (P <0.05) in algal growth. Daily concentrations of algae exposed to varying levels of ozone were compared to the control using Dunnett's t-test.
- Validity criteria fulfilled:
- not applicable
- Remarks:
- no guideline study
- Conclusions:
- The effects of ozone exposure varied among the different species of algae, with C. gracilis appearing the most sensitive and I. galbana the most resistant. Reinocculation showed no residual effects of the treatment. The treatment was limited to an exposure to total residual oxidants varying from species to species, with an approximate range of 0.5 to 3 min. The most sensitive LOEC was determined with 0.06 TRO/L for 0.5 min of treatment, measured one day after exposition for C. gracilis.
- Executive summary:
The effects of ozone treatment on three unicellular algal species, Isochris galbana, Nannochloropsis oculata and Chaetoceros gracilis were studied. I. galbana and N. oculata were found to be resistant to total residual oxidants (TRO), while C. gracilis was relatively sensitive to TRO. I. galbana remained in log-phase growth after treatment with 0.05 to 0.50 mg/L and N. oculata exhibited strong growth after exposure to TRO levels up to 0.67 mg/L. TRO levels of 0.92 and 0.90 mg/L reduced cell counts of N. oculata and I.galbana. C. gracilis initialy showed reductions of cell count at TRO levels above 0.06 mg/L. Log-phase growth was resumed 2d after exposure to TRO as high as 0.31 mg/L.
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- supporting study
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- unsuitable test system
- Qualifier:
- no guideline followed
- Principles of method if other than guideline:
- Samples of swamp water were ozonized for one-half hour and then examined microscopically for percentage removal of living organisms and organic matter.
- GLP compliance:
- no
- Analytical monitoring:
- no
- Details on sampling:
- n/a
- Vehicle:
- no
- Details on test solutions:
- n/a
- Test organisms (species):
- other: see details on test organisms
- Details on test organisms:
- Diatomaceae, Cyanophyceae, Chlorophyceae, Fungi, Protozoa, Rotifera, Crustacea
- Test type:
- flow-through
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 30 min
- Post exposure observation period:
- 0 and 24 hours after treatment samples were examined.
- Hardness:
- no data
- Test temperature:
- no data
- pH:
- no data
- Dissolved oxygen:
- no data
- Salinity:
- no data
- Conductivity:
- no data
- Nominal and measured concentrations:
- Nominal: air with a low ozone concentration (0.2 to 0.3 g per cubic meter) was bubbled through the water sample at a rate of 1 L/min.
- Details on test conditions:
- Ozone containing air was drawn at a rate of 1 L/min through a bottle containing 0.5 L of swamp water. The percentage removal of living organisms and organic matter was examined microscopically.
- Reference substance (positive control):
- no
- Duration:
- 30 min
- Dose descriptor:
- other: percentage of mortality
- Effect conc.:
- other: 0.2 to 0.3 g per cubic meter) lead into the sample water for 30 min with a rate of 1 L/min
- Nominal / measured:
- estimated
- Conc. based on:
- test mat.
- Basis for effect:
- other: removal of organisms
- Details on results:
- Average percentage removal of organisms immediately after treatment:
Diatomaceae 48.0%
Cyanophyceae 55.0%
Chlorophyceae 64.0%
Fungi 46.2%
Protozoa 41.5%
Rotifera 32.8%
Crustacea 30.0% - Validity criteria fulfilled:
- not applicable
- Remarks:
- no guideline study
- Conclusions:
- Ozone affects a broad range of aquatic organisms amongst which are Cyanophycea and Chlorophycea. No observations of effects on algae after 24 hours are reported. Insufficient data are presented to draw firm conclusions.
- Executive summary:
Certain forms of algae are very readily removed from water by direct oxidation while certain other forms are entirely unaffected even with protracted periods of contact with the ozone. In addition to this it has been noted that there is an increase in the odor arising from direct ozonization of algae that can only be removed after complete oxidation of the organic content of the water.
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Data waiving:
- study technically not feasible
- Justification for data waiving:
- other:
- Justification for type of information:
- Ozone is unstable and evaporates from the water phase, hence a flow-through study design would be necessary to maintain stable ozone test concentrations. However, the standard testing methods for algae are limited to static designs (OECD guideline 201: Freshwater Alga and Cyanobacteria, Growth Inhibition Test). A flow-through test design for algal toxicity tests has not yet been developed to date. Therefore, a full-scale guideline compliant algal growth inhibition test (72 h exposure) appears to be technically infeasible. In view of the very short half-life of ozone, the existing supportive studies (although of reliability), along with the fish toxicity data, are considered to be sufficient for ecotoxicological hazard characterisation of ozone.
Referenceopen allclose all
Table 1: Cell counts of Nannochloropsis oculata (x106cells/mL) exposed to varying levels of ozone. Values represent mean (of four replicates. Values within a column having a * are significantly different from the control, using the Dunnett's t-test (P < 0.05).
Missing values (-) were below detectable limits.
Nannochloropsis oculata |
||||||
|
Cell counts (x 106/ml) |
|||||
|
|
|||||
Day after treatment |
d0 |
d1 |
d2 |
d3 |
d4 |
d5 |
TRO in mg/ L (treatment duration in minutes) |
|
|
|
|
|
|
Control |
0.79 |
2.00 |
3.86 |
9.35 |
15.40 |
19.20 |
0.05 (0.5) |
0.79 |
1.81* |
3.42* |
8.72 |
14.46 |
18.13 |
0.12 (0.75) |
0.79 |
1.66* |
3.11* |
7.66* |
13.61 |
18.21 |
0.24 (1) |
0.79 |
1.42* |
2.50* |
6.22* |
12.09* |
15.98 |
0.67 (1.5) |
0.79 |
1.11* |
1.60* |
4.00* |
8.14* |
13.11* |
0.92 (2) |
0.79 |
0.74* |
0.47* |
1.16* |
1.74* |
4.44* |
Pooled Standard Error |
|
0.037 |
0.094 |
0.026 |
0.077 |
0.013 |
|
|
|
|
|
|
|
Table 2: Cell counts of Isochrysis galbana (x106cells/mL) exposed to varying levels of ozone. Values represent mean (of four replicates. Values within a column having a * are significantly different from the control, using the Dunnett's t-test (P < 0.05).
Missing values (-) were below detectable limits.
Isochrysis galbana |
|||||
|
Cell counts (x 106/ml) |
||||
|
|
||||
Day after treatment |
d0 |
d1 |
d2 |
d3 |
d4 |
TRO in mg/ L (treatment duration in minutes) |
|
|
|
|
|
Control |
0.32 |
1.22 |
2.71 |
3.65 |
5.10 |
0.05 (0.5) |
0.32 |
1.16 |
2.82 |
3.74 |
5.22 |
0.23 (1) |
0.32 |
1.03* |
2.63 |
3.40 |
4.96 |
0.34 (1.5) |
0.32 |
0.71* |
2.27* |
3.22* |
4.85 |
0.50 (2) |
0.32 |
0.52* |
1.95* |
2.95* |
4.45* |
0.90 (3) |
0.32 |
0.0.22* |
1.01* |
1.95* |
3.47* |
Pooled Standard Error |
|
0.047 |
0.110 |
0.095 |
0.121 |
|
|
|
|
|
|
Table 3: Cell counts of Chaetoceros gracilis (x106cells/mL) exposed to varying levels of ozone. Values represent mean (of four replicates. Values within a column having a * are significantly different from the control, using the Dunnett's t-test (P < 0.05).
Missing values (-) were below detectable limits.
Chaetoceros gracilis |
||||||
|
Cell counts (x 106/ml) |
|||||
|
|
|||||
Day after treatment |
d0 |
d1 |
d2 |
d3 |
d4 |
d5 |
TRO in mg/ L (treatment duration in minutes) |
|
|
|
|
|
|
Control |
0.31 |
1.14 |
3.74 |
4.54 |
5.41 |
5.77 |
0.06 (0.5) |
0.31 |
0.51* |
1.76* |
4.10 |
5.12 |
5.91 |
0.21 (1) |
0.31 |
0.04* |
0.50* |
2.52* |
4.26 |
4.65 |
0.31 (1.5) |
0.31 |
0.01* |
0.11* |
0.89* |
3.50* |
4.71 |
0.56 (2) |
0.31 |
- |
- |
0.13* |
1.55* |
3.32* |
0.92 (3) |
0.31 |
- |
- |
- |
0.38* |
2.03* |
Pooled Standard Error |
|
0.048 |
0.125 |
0.356 |
0.347 |
0.369 |
Description of key information
Ozone is unlikely to be found in the water phase, as it evaporates immediately from the water phase in to the air or it reacts with other molecules, hence being not stable in water. Therefore, a flow-through study design would be necessary to keep up a defined concentration of ozone. Standard testing methods for algae are limited to static designs. Based on the toxicity in fish (see section 6.1), ozone is already classified in category 1 for chronic aquatic toxicity, which is the most stringent classification for this end-point. Therefore, additional testing of toxicity to aquatic algae or cyanobacteria would not be of added value. Available data from non-standardized tests in scientific publications are presented, showing that ozone at concentrations of 60 µg/L results in significant toxicity after an exposure for between seconds to minutes. A classification to category 1 for acute and chronic aquatic toxicity is reasonable.
Due to the unstable characteristics of ozone, additional testing of toxicity to aquatic algae or cyanobacteria would not be of added value. Available data from non-standardized tests in scientific publications are presented, showing that ozone in concentrations in 60 µg/L already displays significant toxicity after exposure in the range of seconds to minutes. A classification to category 1 for acute and chronic aquatic toxicity is reasonable.
Classification proposal (REGULATION (EC) No 1272/2008 and amendments): Acute and chronic aquatic toxicity category 1.
Key value for chemical safety assessment
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
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