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EC number: 848-536-1 | CAS number: 28911-49-1
- 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:
- 2020-06-08 to 2020-06-11
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Version / remarks:
- Annex 5 corrected 28 July 2011
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- 01 March 2016
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- - Sampling method: For determination of the test item concentrations in the test solutions, all three test vessels were analysed at the start and at the end of the study. In case of control sample, all six test vessels were analysed at the start and at the end of the study.
- Sample storage conditions before analysis: no - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test solutions used in the test was prepared by mechanical dispersion without using of any solubilising agent.
For preparation of test item solutions a stock solution of 100 mg/L (nominal concentration) was first prepared by dissolving an amount of 0.0303 g test item in 303 mL OECD medium. The test solutions of subsequent lower concentrations were prepared by appropriate dilution of this stock solution.
- Controls: negative (blank) control, positive control
- Evidence of undissolved material: no - Test organisms (species):
- Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Strain: 61.81 SAG (identical strains: CCAP 278/4; UTEX 1648; ATCC 22662)
- Source: The algae were supplied by the SAG: Collection of Algal Cultures, Inst. Plant Physiology, University of Göttingen, Untere Karspüle 2, D-37073 Göttingen, Germany.
- Age of inoculum: 3 days (exponentially growth)
- Method of cultivation: The stock cultures are small algal cultures that are planted on agar. These are transferred to fresh medium at least once every two months under standardised conditions according to the test guidelines. The pre-culture was prepared with OECD Medium, incubated under the conditions of the test and used when still exponentially growing, normally after an incubation period of 2-4 days. (The pre-culture was incubated for three days at this test.) - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- None
- Hardness:
- not specified
- Test temperature:
- 22.7 – 23.2 °C
- pH:
- 7.81 – 9.18
- Dissolved oxygen:
- not specified
- Salinity:
- not relevant
- Conductivity:
- not specified
- Nominal and measured concentrations:
- nominal: 0 (blank control), 0.26, 0.64, 1.6, 4.0 and 10 mg/L
measured: 0.18, 0.52, 1.4, 3.7 and 9.3 mg/L (geometric mean) - Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Type: closed with permeable stoppers
- Material, size, fill volume: glass, ~250 mL nominal volume,100 mL fill volume
- Aeration: agitated by using an orbital shaker
- Initial cells density: 10^4 cells/mL in each test flask
- Control end cells density: 10^4 cells/mL
- No. of vessels per concentration: 3
- No. of vessels per control: 6
GROWTH MEDIUM
- Standard medium used: yes (OECD Medium)
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted algal growth medium (OECD medium, according to OECD 201) prepared with deionised water
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: continous illumination
- Light intensity and quality: 7112 lux, wavelength range: 400 – 700 nm
DETERMINATION OF CELL CONCENTRATION
- Determination of cell concentrations: counting chamber
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5
- Range finding study: performed
- Test concentrations: 0 (control), 0.1, 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 1.92 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 0.74 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.52 mg/L
- Nominal / measured:
- meas. (arithm. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 0.95 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 0.43 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.52 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 1.03 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 1.4 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 0.57 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 0.95 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Details on results:
- - Exponential growth in the control: yes
- Observation of abnormalities: no
- Unusual cell shape: No morphological abnormalities were observed in the control and in the concentrations of 0.18, 0.52 mg/L (measured) during the test.
Swollen cells, thin cells or pinches on cells were observed in the concentrations of 1.4, 3.7 and 9.3 mg/L (measured).
- Colour differences: no
- Flocculation: no
- Adherence to test vessels: no
- Aggregation of algal cells: no
- Other: none
- Any stimulation of growth found in any treatment: no
- Any observations that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no
The results of the statistical evaluations showed that the 0 - 72 h average specific growth rate and yield were statistically significantly different from the untreated control at concentrations of 1.4, 3.7 and 9.3 mg/L (measured) and not significantly different at concentrations of 0.18, 0.52 mg/L (measured).
The cell density in the control has increased from nominal N = 1 × 104 cells/mL at the start of the test (0 hours) to 74.67× 104 cells at the end of the test (72 h). - Results with reference substance (positive control):
- 72h ErC50: 0.52 mg/L (95 % confidence limits: 0.46 – 0.58 mg/L)
72h EyC50: 0.31 mg/L (95 % confidence limits: 0.28 – 0.35 mg/L) - Reported statistics and error estimates:
- For determination of the ECx values Probit analysis by SPSS software was performed.
For the determination of the LOEC and NOEC, the calculated mean growth rates and yield at the test concentrations were tested on significant differences to the control values using analysis of variance (ANOVA) and Dunnett’s test (Dunnett T3) by SPSS software. - Validity criteria fulfilled:
- yes
- Conclusions:
- In an algae growth inhibition test according to OECD TG 201 with Raphidocelis subcapitata the 72-hour EC50 and EC10 for the growth rate was determined to be 1.92 and 0.74 mg/L, respectively. The 72-hour EC50 and EC10 based on biomass was found to be 0.95 and 0.43 mg/L, respectively. The 72-hour NOEC was 0.52 mg/L both for the growth rate and biomass.
- Executive summary:
To determine the effect of the test item on the growth of a unicellular green algal species Raphidocelis subcapitata a growth inhibition test according to OECD TG 201 and EU Method C.3. was carried out in compliance with GLP principles. Exponentially growing cultures of Raphidocelis subcapitata were exposed for 72 hours to nominal test item concentration of 0.26, 0.64, 1.6, 4.0 and 10 mg/L over several generations under defined conditions. An untreated control group (OECD Test Medium without test item) was tested parallel with the treated cultures. The test design included three replicates at each test concentration and six replicates for the untreated control. The alga cell concentration was approximately 104 cells/mL at the start of the test in all of the test cultures. The algal cell concentration was determined in 24-hour intervals by manual cell counting using a microscope. The nominal test item concentrations were analytically verified by using a HPLC-UV method. The measured concentrations deviated more than 20 % from the nominal at the two lowest concentrations during the experiment therefore the geometric mean of the measured concentrations were calculated to determine exposure concentrations. The corresponding calculated geometric mean concentrations were the followings: 0.18, 0.52, 1.4, 3.7 and 9.3 mg/L. Biological results and endpoints are thus based on the measured geometric mean concentrations. All validity criteria of the guidelines were met. As a result, the 72-hour EC50 and EC10 for the growth rate was determined to be 1.92 and 0.74 mg/L, respectively. The 72-hour EC50 and EC10 based on biomass was found to be 0.95 and 0.43 mg/L, respectively. The 72-hour NOEC was 0.52 mg/L both for the growth rate and biomass.
Reference
Table 1: Algal cell number (× 104cell/mL) during the test period of 72 hours and Yield
Concentration |
|
Number of Cells (× 104/mL) |
Yield |
||||
nominal |
measured |
0 h |
24 h |
48 h |
72 h |
||
Control |
Not detected |
|
1 |
2 |
17 |
77 |
76 |
|
1 |
3 |
14 |
72 |
71 |
||
|
1 |
3 |
16 |
71 |
70 |
||
|
1 |
2 |
18 |
83 |
82 |
||
|
1 |
2 |
16 |
67 |
66 |
||
|
1 |
2 |
19 |
78 |
77 |
||
Mean |
1.00 |
2.33 |
16.67 |
74.67 |
73.7 |
||
SD |
0.00 |
0.52 |
1.75 |
5.75 |
5.75 |
||
0.26 |
0.18 |
|
1 |
2 |
18 |
74 |
73 |
|
1 |
2 |
20 |
78 |
77 |
||
|
1 |
2 |
14 |
69 |
68 |
||
Mean |
1.00 |
2.00 |
17.33 |
73.67 |
72.7 |
||
SD |
0.00 |
0.00 |
3.06 |
4.51 |
4.51 |
||
0.64 |
0.52 |
|
1 |
2 |
17 |
67 |
66 |
|
1 |
3 |
17 |
74 |
73 |
||
|
1 |
3 |
17 |
71 |
70 |
||
Mean |
1.00 |
2.67 |
17.00 |
70.67 |
69.7 |
||
SD |
0.00 |
0.58 |
0.00 |
3.51 |
3.51 |
||
1.6 |
1.4 |
|
1 |
2 |
11 |
13 |
12 |
|
1 |
2 |
10 |
13 |
12 |
||
|
1 |
2 |
12 |
15 |
14 |
||
Mean |
1.00 |
2.00 |
11.00 |
13.67 |
12.7 |
||
SD |
0.00 |
0.00 |
1.00 |
1.15 |
1.15 |
||
4.0 |
3.7 |
|
1 |
2 |
3 |
4 |
3 |
|
1 |
2 |
2 |
2 |
1 |
||
|
1 |
2 |
4 |
3 |
2 |
||
Mean |
1.00 |
2.00 |
3.00 |
3.00 |
2.0 |
||
SD |
0.00 |
0.00 |
1.00 |
1.00 |
1.00 |
||
10 |
9.3 |
|
1 |
1 |
1 |
1 |
0 |
|
1 |
1 |
1 |
1 |
0 |
||
|
1 |
1 |
1 |
1 |
0 |
||
Mean |
1.00 |
1.00 |
1.00 |
1.00 |
0.0 |
||
SD |
0.00 |
0.00 |
0.00 |
0.00 |
0.00 |
Table 2: The specific Growth Rates during the test period of 72 hours
Concentration |
|
Growth Rates (µ) |
|||
nominal |
measured |
0 – 24 h |
0 – 48 h |
0 – 72 h |
|
Control |
Not detected |
|
0.0289 |
0.0590 |
0.0603 |
|
0.0458 |
0.0550 |
0.0594 |
||
|
0.0458 |
0.0578 |
0.0592 |
||
|
0.0289 |
0.0602 |
0.0614 |
||
|
0.0289 |
0.0578 |
0.0584 |
||
|
0.0289 |
0.0613 |
0.0605 |
||
Mean |
0.0345 |
0.0585 |
0.0599 |
||
SD |
0.0087 |
0.0022 |
0.0011 |
||
0.26 |
0.18 |
|
0.0289 |
0.0602 |
0.0598 |
|
0.0289 |
0.0624 |
0.0605 |
||
|
0.0289 |
0.0550 |
0.0588 |
||
Mean |
0.0289 |
0.0592 |
0.0597 |
||
SD |
0.0000 |
0.0038 |
0.0009 |
||
0.64 |
0.52 |
|
0.0289 |
0.0590 |
0.0584 |
|
0.0458 |
0.0590 |
0.0598 |
||
|
0.0458 |
0.0590 |
0.0592 |
||
Mean |
0.0401 |
0.0590 |
0.0591 |
||
SD |
0.0098 |
0.0000 |
0.0007 |
||
1.6 |
1.4 |
|
0.0289 |
0.0500 |
0.0356 |
|
0.0289 |
0.0480 |
0.0356 |
||
|
0.0289 |
0.0518 |
0.0376 |
||
Mean |
0.0289 |
0.0499 |
0.0363 |
||
SD |
0.0000 |
0.0019 |
0.0011 |
||
4.0 |
3.7 |
|
0.0289 |
0.0229 |
0.0193 |
|
0.0289 |
0.0144 |
0.0096 |
||
|
0.0289 |
0.0289 |
0.0153 |
||
Mean |
0.0289 |
0.0221 |
0.0147 |
||
SD |
0.0000 |
0.0073 |
0.0048 |
||
10 |
9.3 |
|
0.0000 |
0.0000 |
0.0000 |
|
0.0000 |
0.0000 |
0.0000 |
||
|
0.0000 |
0.0000 |
0.0000 |
||
Mean |
0.0000 |
0.0000 |
0.0000 |
||
SD |
0.0000 |
0.0000 |
0.0000 |
Table 3: Growth Rates (µ) and percentage inhibition of µ during the test period
Concentration |
Growth Rates (µ)and % Inhibition ofµ |
||||||
nominal |
measured |
0–24 h |
0–48 h |
0–72 h |
|||
µ |
% |
µ |
% |
µ |
% |
||
Control |
Not detected |
0.0345 |
– |
0.0585 |
– |
0.0599 |
– |
0.26 |
0.18 |
0.0289 |
16.3 |
0.0592 |
-1.2 |
0.0597 |
0.3 |
0.64 |
0.52 |
0.0401 |
-16.3 |
0.0590 |
-0.9 |
0.0591 |
1.2 |
1.6 |
1.4 |
0.0289 |
16.3 |
0.0499 |
14.7 |
0.0363 |
39.4 |
4.0 |
3.7 |
0.0289 |
16.3 |
0.0221 |
62.3 |
0.0147 |
75.4 |
10 |
9.3 |
0.0000 |
100.0 |
0.0000 |
100.0 |
0.0000 |
100.0 |
Table 4: Morphological changes of algal cells observed after 24, 48 and 72 hours of exposure
Concentration |
Morphological changes |
|||
nominal |
measured |
24 h |
48 h |
72 h |
Control |
Not detected |
- normal cells |
- normal cells |
- normal cells |
0.26 |
0.18 |
- normal cells |
- normal cells |
- normal cells |
0.64 |
0.52 |
- normal cells |
- normal cells |
- normal cells |
1.6 |
1.4 |
- normal cells |
- normal cells |
- swollen cells |
4.0 |
3.7 |
- normal cells |
- swollen cells |
- swollen cells |
10 |
9.3 |
- swollen cells |
- swollen cells |
- swollen cells |
Table 5: Concentration of Test Item measured in the Test Solutions
Nominal concentration mg/L |
Mean of the measured concentrations |
|||
Start(June 08, 2020) |
End(June 11, 2020) |
|||
Concentration (mg/L) |
% of the nominal |
Concentration (mg/L) |
% of the nominal |
|
Control |
not detected |
- |
not detected |
- |
0.26 |
0.230 |
89 |
0.140 |
54 |
0.64 |
0.597 |
93 |
0.453 |
71 |
1.6 |
1.477 |
92 |
1.381 |
86 |
4 |
3.702 |
93 |
3.654 |
91 |
10 |
9.274 |
93 |
9.353 |
94 |
Description of key information
In an algae growth inhibition test according to OECD TG 201 with Raphidocelis subcapitata the 72-hour EC50 and EC10 for the growth rate was determined to be 1.92 and 0.74 mg/L, respectively. The 72-hour EC50 and EC10 based on biomass was found to be 0.95 and 0.43 mg/L, respectively. The 72-hour NOEC was 0.52 mg/L both for the growth rate and biomass.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 1.92 mg/L
- EC10 or NOEC for freshwater algae:
- 0.74 mg/L
Additional information
To determine the effect of the test item on the growth of a unicellular green algal species Raphidocelis subcapitata a growth inhibition test according to OECD TG 201 and EU Method C.3. was carried out in compliance with GLP principles. Exponentially growing cultures of Raphidocelis subcapitata were exposed for 72 hours to nominal test item concentration of 0.26, 0.64, 1.6, 4.0 and 10 mg/L over several generations under defined conditions. An untreated control group (OECD Test Medium without test item) was tested parallel with the treated cultures. The test design included three replicates at each test concentration and six replicates for the untreated control. The alga cell concentration was approximately 104cells/mL at the start of the test in all of the test cultures. The algal cell concentration was determined in 24-hour intervals by manual cell counting using a microscope. The nominal test item concentrations were analytically verified by using a HPLC-UV method. The measured concentrations deviated more than 20 % from the nominal at the two lowest concentrations during the experiment therefore the geometric mean of the measured concentrations were calculated to determine exposure concentrations. The corresponding calculated geometric mean concentrations were the followings: 0.18, 0.52, 1.4, 3.7 and 9.3 mg/L. Biological results and endpoints are thus based on the measured geometric mean concentrations. All validity criteria of the guidelines were met. As a result, the 72-hour EC50 and EC10 for the growth rate was determined to be 1.92 and 0.74 mg/L, respectively. The 72-hour EC50 and EC10 based on biomass was found to be 0.95 and 0.43 mg/L, respectively. The 72-hour NOEC was 0.52 mg/L both for the growth rate and biomass.
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