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EC number: 218-780-8 | CAS number: 2234-16-4
- 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:
- key study
- Study period:
- From 26 March 2018 to 29 March 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 850.5400 (Algal Toxicity, Tiers I and II) (January 2012)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: 6.25, 12.5, 25, 50 and 100 % saturated solutions.
- Sampling method: Samples were taken from the test solutions at the applied concentration levels and from the control solution at the beginning and at the end of the experiment.
- Sample storage conditions before analysis: not required. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Because the Test Item was not readily soluble in water by direct addition, a test solution was prepared using a saturated solution method. The solution was prepared by dispersing/dissolving 100 mg/L Test Item into the test medium (OECD Medium) two days before the start of the experiment. This solution was shaken for approximately 24 hours at approximately 30°C (Day - 2) and then was equilibrated for approximately 24 hours at approximately 20°C (Day - 1). Any non-dissolved test materials were removed by filtration through a fine (0.22 μm) filter to give the 100% saturated solution. The analysis of the prepared solution showed a solution of 93.3 mg/L, so the method was considered valid for the study.
- Differential loading: nominal loading rate of 100 mg/L
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): no. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Specie: Pseudokirchneriella subcapitata
- Strain: 61.81 SAG (identical strains: CCAP 278/4; UTEX 1648; ATCC 22662)
- Source (laboratory, culture collection): The algae were supplied by the SAG: Collection of Algal Cultures, Inst. Plant Physiology, University of Göttingen, Germany.
- Method of cultivation: The algae were cultured under standarised conditions, according to OECD 201 in the Ecotoxicological Laboratory of Citoxlab Hungary Ltd. The pre-culture, which is intended to give a quantity of algae suitable for the inoculation of test cultures, was prepared with the OECD algal growth medium, incubated under the same conditions as the test and used when still exponentially growing (after an incubation period of 3 days). If the algal cultures contained deformed or abnormal cells, they were discarded. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 72 h
- Test temperature:
- 22.5 – 22.8 °C
- pH:
- 7.32 – 8.71
- Nominal and measured concentrations:
- Nominal concentrations: 0 (control), 6.25, 12.5, 25, 50 and 100% saturated solutions.
Measured concentrations: 0 (control), 4.59, 9.75, 19.6, 42.1 and 79.9 mg/L (geometric mean). - Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Material, size, headspace, fill volume: 250 mL Erlenmeyer flasks, continuously shaken by a laboratory orbital shaker to keep algae in suspension. The flasks were covered with air-permeable stoppers.
- Aeration: No
- Initial cells density: 1E+04 algal cells per mL test medium
- Control end cells density: The cell density in the control cultures increased by the factor of 69.0 within three days.
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS: Reconstituted algal growth medium (OECD medium, according to OECD 201) was used as dilution water for both the range finding and definitive tests.
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: No
- Photoperiod: Continuously illuminated.
- Light intensity and quality: 7594 lux (equivalent to ~103 μE/m2/s), which was ensured with fluorescent lamps (with a spectral range of 400-700 nm). The differences in light intensity between the test vessels did not exceed ± 15 % and therefore provided equal conditions for each test vessel.
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The cell numbers were determined at 24, 48 and 72 hours after starting the test by manual cell counting using a microscopic method with a counting chamber. Microscopic observation of the algal cells in test concentration level and in the control was performed (at 24h, 48h and 72h) to detect any abnormal appearance of the algae.
TEST CONCENTRATIONS
Range finding study: A concentration range-finding test was conducted to determine the approximate toxicity of the test item so that appropriate test concentrations could be selected for use in the definitive test.
- Test concentrations: 0, 0.1, 1, 10 and 100 mg/L (2 replicates per each test concentration and 3 replicates in the control group, 72 hours)
- Results used to determine the conditions for the definitive study: At the highest applied concentration (100% saturated solution) chuff cells were observed. In the other applied test concentrations all cells were normal. Because a toxic response was observed during the preliminary range-finding test, five test concentrations in a geometric series (factor 2.0) and one Control were tested in the main experiment. The concentrations in the definitive test were based on the results of the preliminary range-finding test and were: 6.25, 12.5, 25, 50 and 100 % saturated solutions. - Reference substance (positive control):
- yes
- Remarks:
- (Potassium dichromate is tested at least twice a year by CitoxLab)
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 14.51 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95% CL: 13.16 – 15.99 mg/L
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 4.59 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 9.21 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95% CL: 8.43 –10.06 mg/L
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 4.59 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 8.87 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Remarks on result:
- other: 95% CL: 8.19 – 9.60 mg/L
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 4.59 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield
- Details on results:
- - Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): Chuff cells were observed at the tested concentration of 9.75, 19.6, 42.1 and 79.9 mg/L. There was no morphological deviation observed in the tested concentrations of 4.59 mg/L and in the Control during the study, all these cells were normal.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no.
- Effect concentrations exceeding solubility of substance in test medium: no. - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
Potassium dichromate was tested at least twice a year to demonstrate satisfactory test conditions.
The date of the last study (Study Code: 17/367-022AL) with the reference item Potassium dichromate is (Batch Number: A0345704): 04-07/12/2017
The 72h ErC 50: 0.88 mg/L, (95 % confidence limits: 0.81 – 0.96 mg/L)
The 72h EbC 50: 0.63 mg/L, (95 % confidence limits: 0.58 – 0.69 mg/L)
The 72h EyC 50: 0.53 mg/L, (95 % confidence limits: 0.49 – 0.58 mg/L)
These values are within the range of laboratory ring test data. - Reported statistics and error estimates:
- The section-by-section specific growth rates in the control cultures were assessed (calculated as the specific growth rates for each day during the course of the test, days 0-1, 1-2 and 2-3) and to demonstrate exponential growth for the entire study period. The inhibition of alga growth was determined from the biomass (area under the growth curves, A), the average specific growth rate (r) and from the yield (y). Mean values and standard deviations were calculated for each concentration at the start, and at the end of the test using Excel for Windows software (Microsoft Co./One Microsoft Way/Redmond, WA 98052-6399).
The ErC10, 20, 50, EbC10, 20, 50 and EyC10, 20, 50 values of the Test Item and their confidence limits were calculated using Probit analysis by TOXSTAT software. Statistical comparisons of biomass, average specific growth rates and yield in controls and in the treated groups were carried out using analysis of variance (ANOVA) and Bonferroni t-Test (α = 0.05) by TOXSTAT software. For the determination of the LOEC and NOEC, the calculated mean biomass, growth rates and yield at the test concentrations were tested on significant differences to the control values by Bonferroni t-Test. - Validity criteria fulfilled:
- yes
- Remarks:
- (cell density in control increased by the factor of 69.0>16, mean coefficient of variation for section-by-section specific growth rates in control was 13.24%<35%, coefficient of variation of average specific growth rates in control was 1.3%<7%)
- Conclusions:
- The 72h-EC50 in green algae Pseudokirchneriella subcapitata was found to be 14.51 mg/L (measured, geometric mean; 95% CL: 13.16 – 15.99 mg/L), the EC10 was 6.70 mg/L (measured, geometric mean; 95% CL: 5.73 – 7.85 mg/L) and the NOEC was determined to be 4.59 mg/L, based on growth rate.
- Executive summary:
A Freshwater Alga Growth Inhibition Test was performed in accordance with OECD Guideline 201 and EU Method C.3, following GLP. The effect of test item was assessed on algal growth using the unicellular green alga Pseudokirchneriella subcapitata, over an exposure period of 72 hours in a static system. Based on the results of a preliminary range-finding test, algae cultures were exposed to 6.25, 12.5, 25, 50 and 100% saturated solutions of the test item (due to low water solubility) or a control. The test design included three replicates per test concentration and six replicates for the untreated control. Test concentrations were analytically determined at the beginning and at the end of the test by an HPLC-UV method. As the deviation from the initial values was greater than 20% from the nominal, measured (geometric mean) concentrations are reported: 4.59, 9.75, 19.6, 42.1 and 79.9 mg/L test item. Positive controls (postassium dichromate) are performed at least twice a year to ensure the quality of the results. All validity criteria were met. The 72h-EC50 in green algae Pseudokirchneriella subcapitata was found to be 14.51 mg/L (measured, geometric mean; 95% CL: 13.16 – 15.99 mg/L), the EC10 was 6.70 mg/L (measured, geometric mean; 95% CL: 5.73 – 7.85 mg/L) and the NOEC was determined to be 4.59 mg/L, both based on growth rate.
Reference
Table 2. Summary of results.
Parameter |
Growth rate (r) |
Yield (y) |
Biomass (b) |
EC10 |
6.70 |
5.38 |
4.96 |
95 % conf. limits |
5.73 – 7.85 |
4.75 –6.09 |
4.30 – 5.73 |
EC20 |
8.74 |
6.39 |
6.14 |
95 % conf. limits |
7.68 – 9.94 |
5.76 – 7.08 |
5.45 – 6.90 |
EC50 |
14.51 |
8.87 |
9.21 |
95 % conf. limits |
13.16 – 15.99 |
8.19 – 9.60 |
8.43 –10.06 |
NOEC |
4.59 |
4.59 |
4.59 |
LOEC |
9.75 |
9.75 |
9.75 |
Table 3. Calculation of the exposure concentrations.
Nominal concentrations (% saturated) |
Measured concentrations |
Geometric mean |
|
0h |
72h |
||
Control |
n.d. |
n.d. |
--- |
6.25 |
5.91 |
3.57 |
4.59 |
12.5 |
11.8 |
8.05 |
9.75 |
25 |
23.6 |
16.3 |
19.6 |
50 |
47.3 |
37.4 |
42.1 |
100 |
93.3 |
68.4 |
79.9 |
Table 4. Growth Rates (µ) and Percentage Inhibition of µ during the Test Period.
Concentrations |
Growth ratemand % inhibition ofm |
||||||
Nominal (% saturated) |
Geometric Mean Measured (mg/L) |
0–24 h |
0–48 h |
0–72 h |
|||
m |
% |
m |
% |
m |
% |
||
Control |
n.d. |
0.0589 |
0.0 |
0.0611 |
0.0 |
0.0588 |
0.0 |
6.25 |
4.59 |
0.0569 |
3.4 |
0.0601 |
1.6 |
0.0586 |
0.3 |
12.5 |
9.75 |
0.0458 |
22.2 |
0.0424* |
30.6 |
0.0450* |
23.5 |
25 |
19.6 |
0.0193* |
67.3 |
0.0144* |
76.4 |
0.0096* |
83.6 |
50 |
42.1 |
0.0096* |
83.6 |
0.0096* |
84.2 |
0.0064* |
89.1 |
100 |
79.9 |
0.0000* |
100.0 |
0.0000* |
100.0 |
0.0000* |
100.0 |
*statistically significantly different compared to the control values (Bonferroni t-Test; α = 0.05).
Table 5. Area under the Growth Curves (A) and Percentage Inhibition of A during the Test Period
Concentration |
Growth ratemand % inhibition ofm |
||||||
Nominal (% saturated) |
Geometric Mean Measured (mg/L) |
0–24 h |
0–48 h |
0–72 h |
|||
m |
% |
m |
% |
m |
% |
||
Control |
n.d |
38.0 |
0.0 |
290.0 |
0.0 |
1320.0 |
0.0 |
6.25 |
4.59 |
36.0 |
5.3 |
276.0 |
4.8 |
1284.0 |
2.7 |
12.5 |
9.75 |
24.0 |
36.8 |
128.0* |
55.9 |
504.0* |
61.8 |
25 |
19.6 |
8.0* |
78.9 |
28.0* |
90.3 |
52.0* |
96.1 |
50 |
42.1 |
4.0* |
89.5 |
16.0* |
94.5 |
32.0* |
97.6 |
100 |
79.9 |
0.0* |
100.0 |
0.0* |
100.0 |
0.0* |
100.0 |
*statistically significantly different compared to the control values (Bonferroni t-Test;a= 0.05).
Table 6. Yield (Y) and Percentage Inhibition of Y during the Test Period
Concentration |
Yield (Y) and Percentage Inhibition |
||||
Nominal (% saturated) |
Geometric Mean Measured (mg/L) |
0–72 h |
|||
Y |
% |
||||
Control |
n.d |
68.0 |
0.0 |
||
6.25 |
4.59 |
67.0 |
1.5 |
||
12.5 |
9.75 |
24.7* |
63.7 |
||
25 |
19.6 |
1.0* |
98.5 |
||
50 |
42.1 |
0.7* |
99.0 |
||
100 |
79.9 |
0.0* |
100.0 |
||
*statistically significantly different compared to the control values (Bonferroni t-Test;a= 0.05).
Table 7. Influence of Test Item on the Growth ofPseudokirchneriella subcapitata
Parameter |
Growth rate (r) |
Yield (y) |
Biomass (b) |
EC10 |
6.70 |
5.38 |
4.96 |
95 % conf. limits |
5.73 – 7.85 |
4.75 –6.09 |
4.30 – 5.73 |
EC20 |
8.74 |
6.39 |
6.14 |
95 % conf. limits |
7.68 – 9.94 |
5.76 – 7.08 |
5.45 – 6.90 |
EC50 |
14.51 |
8.87 |
9.21 |
95 % conf. limits |
13.16 – 15.99 |
8.19 – 9.60 |
8.43 –10.06 |
NOEC |
4.59 |
4.59 |
4.59 |
LOEC |
9.75 |
9.75 |
9.75 |
Table 8. pH-Values in the Test Media at the Start and End of the Test
Concentration |
pH-values |
||
Nominal |
Measured |
Start |
End |
Control |
n.d. |
7.38 |
8.71 |
8.64 |
|||
8.65 |
|||
8.70 |
|||
8.63 |
|||
8.67 |
|||
6.25 |
4.59 |
7.38 |
8.52 |
8.50 |
|||
8.62 |
|||
12.5 |
9.75 |
7.37 |
8.07 |
8.05 |
|||
7.95 |
|||
25 |
19.6 |
7.34 |
7.85 |
7.86 |
|||
7.87 |
|||
50 |
42.1 |
7.32 |
7.86 |
7.85 |
|||
7.88 |
|||
100 |
79.9 |
7.34 |
7.82 |
7.84 |
|||
7.82 |
Table 9. Temperature in the Climate Chamber and in the Test Media during the Test
Parameter |
Exposure Time |
||||
Day 0 |
Day 1 |
Day 2 |
Day 3 |
||
temperature (ºC) measured in flask |
22.7 |
22.8 |
22.6 |
22.5 |
|
min / max temperature (ºC) |
min |
22.0 |
22.1 |
22.0 |
22.1 |
max |
23.1 |
23.3 |
22.9 |
23.0 |
Table 10. Cell Number (x 104cell/mL) determined in the Main Experiment
Concentration |
|
Number of cells |
||||
Nominal |
Measured |
|
0 h |
24 h |
48 h |
72 h |
mg/L |
mg/L |
|
||||
Control |
n.d. |
|
1 |
5 |
22 |
73 |
|
1 |
4 |
19 |
63 |
||
|
1 |
5 |
18 |
72 |
||
|
1 |
4 |
19 |
71 |
||
|
1 |
3 |
16 |
68 |
||
|
1 |
4 |
19 |
67 |
||
Mean |
1.00 |
4.17 |
18.83 |
69.00 |
||
SD |
0.0 |
0.8 |
1.9 |
3.7 |
||
6.25 |
4.59 |
|
1 |
4 |
17 |
70 |
|
1 |
3 |
16 |
66 |
||
|
1 |
5 |
21 |
68 |
||
Mean |
1.00 |
4.00 |
18.00 |
68.00 |
||
SD |
0.0 |
1.0 |
2.6 |
2.0 |
||
12.5 |
9.75 |
|
1 |
3 |
7 |
22 |
|
1 |
3 |
8 |
26 |
||
|
1 |
3 |
8 |
29 |
||
Mean |
1.00 |
3.00 |
7.67 |
25.67 |
||
SD |
0.0 |
0.0 |
0.6 |
3.5 |
||
25 |
19.6 |
|
1 |
2 |
2 |
2 |
|
1 |
1 |
2 |
2 |
||
|
1 |
2 |
2 |
2 |
||
Mean |
1.00 |
1.67 |
2.00 |
2.00 |
||
SD |
0.0 |
0.6 |
0.0 |
0.0 |
||
50 |
42.1 |
|
1 |
2 |
2 |
2 |
|
1 |
1 |
2 |
2 |
||
|
1 |
1 |
1 |
1 |
||
Mean |
1.00 |
1.33 |
1.67 |
1.67 |
||
SD |
0.0 |
0.6 |
0.6 |
0.6 |
||
100 |
79.9 |
|
1 |
1 |
1 |
1 |
|
1 |
1 |
1 |
1 |
||
|
1 |
1 |
1 |
1 |
||
Mean |
1.00 |
1.00 |
1.00 |
1.00 |
||
SD |
0.0 |
0.0 |
0.0 |
0.0 |
Description of key information
Key study. Method according to OECD 201 and EU Method C.3. GLP study. The 72h-EC50 in green algae Pseudokirchneriella subcapitata was found to be 14.51 mg/L (measured, geometric mean; 95% CL: 13.16 – 15.99 mg/L), the EC10 was 6.70 mg/L (measured, geometric mean; 95% CL: 5.73 – 7.85 mg/L) and the NOEC was determined to be 4.59 mg/L, based on growth rate.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 14.51 mg/L
- EC10 or NOEC for freshwater algae:
- 4.59 mg/L
Additional information
A Freshwater Alga Growth Inhibition Test was performed in accordance with OECD Guideline 201 and EU Method C.3, following GLP. Based on the results of a preliminary range-finding test, algae cultures of Pseudokirchneriella subcapitata were exposed to 6.25, 12.5, 25, 50 and 100% saturated solutions of the test item or a control, over an exposure period of 72 hours in a static system. The corresponding measured (geometric mean) concentrations were 4.59, 9.75, 19.6, 42.1 and 79.9 mg/L, analytically determined at the beginning and at the end of the test. Positive controls (postassium dichromate) are performed at least twice a year to ensure the quality of the results. All validity criteria were met. The 72h-EC50 in green algae was found to be 14.51 mg/L (measured, geometric mean; 95% CL: 13.16 – 15.99 mg/L), the EC10 was 6.70 mg/L (measured, geometric mean; 95% CL: 5.73 – 7.85 mg/L) and the NOEC was determined to be 4.59 mg/L, based on growth rate.
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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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