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EC number: 473-310-0 | CAS number: 478385-88-5
- 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:
- 2019-12-16 to 2019-12-20
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Version / remarks:
- 2011
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- - Batch number: K51496539
- Analytical monitoring:
- yes
- Details on sampling:
- - Preparation of the samples: The sample and the control were diluted with acetonitrile containing 0.2% ortho phosphoric acid (factor 2) directly after sampling.
- Sample storage: All samples were stored at room temperature until start of analysis, if necessary. Prepared samples were stored at room temperature until analysis. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A saturated solution with a nominal loading rate of 100 mg test item/L was prepared once at room temperature 24 ± 1 hour prior to the start of the exposure. An appropriate amount of the test item was weighed out with an appropriate amount of dilution water. The saturated solution was stirred for 24 ± 1 hours (1100 rpm, room temperature) with a magnetic stirrer. Undissolved particles were removed by membrane filtration (membrane filter 0.45 µm). The filter was saturated in order to avoid adsorption during the filtration. The first 25 mL of the filtrate were discarded. The filtration was interrupted for 15 minutes to allow for adsorption and saturation of the filter material with dissolved test item. Thereafter, the filtration was continued. The next 25 mL were discarded. The following filtrate, i.e. the saturated solution, was used in the test. During filtration, the filter was always kept covered. The saturated solution was checked via laser beam (Tyndall effect) for undissolved test item. The Tyndall effect was negative.
- Differential loading: One limit loading rate was tested.
- Controls: Six replicates (without test item) were exposed under the same conditions as the test concentrations; Potassium dichromate was tested as a reference item
- Evidence of undissolved material: No. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Strain: CCAP 278/4 (axenic)
- Source: Culture Collection of Algae and Protozoa (CCAP), SAMS Research Services Ltd, Dunstaffnage Marine Laboratory Dunbeg, OBAN; Argyll PA37 1QA; Scotland, UK
- Age of inoculum: A four days old preculture, prepared in dilution water, was used as inoculum.
- Method of cultivation: Fresh stocks are prepared every month on Z-Agar. Light intensity amounts 2567 - 5130 lux for 24 hours per day. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 72 h
- Post exposure observation period:
- none
- Hardness:
- 32 mg CaCO3/L
- Test temperature:
- 21.8 °C
- pH:
- 7.88 -9.42
- Dissolved oxygen:
- not specified
- Salinity:
- not applicable
- Conductivity:
- 137 µS/cm
- Nominal and measured concentrations:
- nominal: 100 mg/L
measured: < LOQ - Details on test conditions:
- TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Type: closed, sealed with cotton wool plugs
- Material, size, fill volume: glass, 250 mL, 100 mL
- Aeration: passive; test containers were placed on a rotary shaker and oscillated at approximately 70 rpm
- Initial cells density: approximately 5 x 10^3 - 10^4 cells/mL in nominal, measured to be 5416 cells/mL
- No. of vessels per concentration: 6
- No. of vessels per control: 6
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Ultrapure water was used to prepare the dilution water.
- Conductivity: max. 0.1 µS/cm
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: 24 hours/day light
- Light intensity and quality: Approximately 4440 to 8880 lux, corresponding to 60 to 120 µE*m-2*s-1; light homogeneity within ± 15% over incubation area
EFFECT PARAMETERS MEASURED
- Determination of cell concentrations: counting chamber; fluorimeter
- Chlorophyll measurement: The cell density was measured daily via Chlorophyll a-fluorescence, excitation at 436 nm, emission at 685 nm. Dilution water was used as a background signal. No self-fluorescence was found in the preliminary range finding test at the saturated solution (100 mg/L).
- Microscopic evaluation: Microscopic evaluation of the cells at the start and the end of the incubation period was carried out. The cells were checked for unusual cell shapes, colour differences, differences in chloroplast morphology, flocculation, adherence of algae to test containers and agglutination of algae cells.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable (limit test)
- Range finding study: A non-GLP preliminary range finding test under static conditions over a period of 72 hours with headspace was conducted at the test facility with a saturated solution with a nominal loading rate of 100 mg/L. Out of a saturated solution two dilution levels were prepared with the concentrations 1.00 and 10.0% of the saturated solution. In the range finding test, two replicates per concentration and four for the control were tested. All concentration levels were visually clear throughout the exposure period.
- 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:
- NOELR
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EL10
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOELR
- Effect conc.:
- < 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- EL10
- Effect conc.:
- < 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- LOELR
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EL20
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOELR
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- EL20
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Details on results:
- - Exponential growth in the control: yes, increase by 244-fold (specific growth rate 1.83 day^- 1)
- Observation of abnormalities: no
- Unusual cell shape: no
- Colour differences: no
- Flocculation: no
- Adherence to test vessels: no
- Aggregation of algal cells: no
- 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 - Results with reference substance (positive control):
- ErC50 = 0.940 mg/L (nominal), 95% confidence interval: 0.732 - 1.33
EyC50 = 0.660 mg/L (nominal), 95% confidence interval: 0.393 - 0.700 - Reported statistics and error estimates:
- EL-values of the growth rate and yield inhibition were estimated empirically based on the results of the single treatment level (limit test design). NOEL/LOEL were determined by calculation of statistical significance of growth rate and yield. As a standard, a t-test was used for NOEL/LOEL calculations. When running a t-test a Normality test and an Equal Variance test were done first. The Shapiro-Wilk-Test was used to test for normal distribution of data. The Brown-Forsythe test was done for analysis of variance homogeneity. P-values for both Normality and Equal Variance tests are 0.05. The α-value (acceptable probability of incorrectly concluding that there is a difference) is α=0.05.
- Validity criteria fulfilled:
- yes
- Conclusions:
- In a limit test according to OECD Guideline 201 and EU Method C.3 with the unicellular freshwater green alga Pseudokirchneriella subcapitata, the 72-hour NOEC (growth rate) of the test item was determined to be 100 mg/L in nominal. The 72-hour ErL10 and ErL50 for the growth rate were >100 mg/L (nominal). The 72-hour EyL50 based on the yield was determined to be >100 mg/L (nominal). The 72-hour NOEC (yield) and EyL10 (yield) were found to be below 100 mg/L (nominal).
- Executive summary:
The toxicity of the test item to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined in a limit-test according to the principles of OECD 201 and Council Regulation (EC) No. 266/2016/Method C.3. The study was conducted over a period of 72 hours under static conditions with an initial cell density of 5416 cells/mL. A saturated solution with a nominal loading rate of 100 mg test item/L was prepared once at room temperature 24 ± 1 hour prior to the start of the exposure. The saturated solution was stirred for 24 ± 1 hours (1100 rpm, room temperature) with a magnetic stirrer. Undissolved particles were removed by membrane filtration. The saturated solution was checked via laser beam (Tyndall effect) for undissolved test item. The Tyndall effect was negative and no undissolved test material was hence detected. Six replicates were tested for the limit loading and six for the control. The test media were clear throughout the test period. The environmental conditions were within the acceptable limits and all validity criteria of the test guidelines were fulfilled. The limit loading rate and the control were analytically verified by HPLC-DAD at the start and the end of the exposure. The measured concentrations of the saturated solution of the test item in the fresh and in the old media were below the Limit of Quantity (LOQ). All effect values given are thus based on the nominal loading of the test item. In result, the 72-hour NOEL for the growth rate was determined to be 100 mg/L. The differences of growth rate were calculated to be statistically significant, but considered biologically not relevant since the inhibition was below 5%. Consequently, the 72-hour ErL10 and ErL50 were determined to be >100 mg/L. The 72-hour NOEL (yield) and EyL10 (yield) were found to be <100 mg/L. The 72-hour EyL50 was determined to be >100 mg/L.
Reference
Table 1: Cell densities.
Nominal test item loading [mg/L] | Replicate No. | Cell denisty [cells/mL] | |||
0 hours | 24 hours | 48 hours | 72 hours | ||
100 mg/L | 1 | 5416 | 16320 | 129219 | 969007 |
2 | 5416 | 17676 | 134321 | 1057848 | |
3 | 5416 | 19046 | 139257 | 1089636 | |
4 | 5416 | 16113 | 152284 | 1072842 | |
5 | 5416 | 17513 | 152936 | 1173950 | |
6 | 5416 | 17878 | 149168 | 1172928 | |
Mean | 5416 | 17424 | 142864 | 1089369 | |
Control | 1 | 5416 | 23131 | 1740.34 | 1324712 |
2 | 5416 | 21210 | 162915 | 1263473 | |
3 | 5416 | 22301 | 171619 | 1314733 | |
4 | 5416 | 22841 | 180849 | 1374853 | |
5 | 5416 | 20499 | 175884 | 1284502 | |
6 | 5416 | 23525 | 173956 | 1363023 | |
Mean | 5416 | 22251 | 173210 | 1320883 |
Table 2: Evaluation after 72 hours. Statistically significant differences
of growth rates and yield compared to control values are marked (+),
not significant differences are marked (-).
Nominal test item loading [mg/L] | Replicate No. | Growth rate [d-1] | Growth rate inhibition [%] | Yield [cells/mL] | Inhibition of yield [%] |
100 | 1 | 1.73 | 6 | 963591 | 27 |
2 | 1.76 | 4 | 1052432 | 20 | |
3 | 1.77 | 3 | 1084220 | 18 | |
4 | 1.76 | 4 | 1067426 | 19 | |
5 | 1.79 | 2 | 1168534 | 11 | |
6 | 1.79 | 2 | 1167512 | 11 | |
Mean | (+)* 1.77 | 4 | (+) 1083953 | 18 | |
Control | 1 | 1.83 | 1319296 | ||
2 | 1.82 | 1258057 | |||
3 | 1.83 | 1309317 | |||
4 | 1.85 | 1369437 | |||
5 | 1.82 | 1279086 | |||
6 | 1.84 | 1357607 | |||
Mean | 1.83 | 1315467 |
* = statistically significant but biologically not relevant (inhibition < 5%)
Table 3: Section-by-Section and Average Specific Growth Rates of the Control Group (0 -72 hours).
Replicate No. | Specific growth rate [d1] section-by-section | Mean (0 - 72 hours) | SD± | CV [%] | Mean CV [%] | |||
0 - 24 hours | 24 - 48 hours | 48 - 72 hours | ||||||
Control | 1 | 1.45 | 2.02 | 2.03 | 1.83 | 0.33 | 18 | 20 |
2 | 1.37 | 2.04 | 2.05 | 1.82 | 0.392 | 21.6 | ||
3 | 1.42 | 2.04 | 2.04 | 1.83 | 0.360 | 19.7 | ||
4 | 1.44 | 2.07 | 2.03 | 1.85 | 0.353 | 19.1 | ||
5 | 1.33 | 2.15 | 1.99 | 1.82 | 0.434 | 23.8 | ||
6 | 1.47 | 2 | 2.06 | 1.84 | 0.325 | 17.6 | ||
Mean | 1.83 | |||||||
SD ± | 0.01 | |||||||
CV [%] | 0.6 |
SD = Standard deviation
CV = Coefficient of variation
Table 4: Analytical results.
Sampling date | Fresh Media 0 hours | Old Media 72 hours |
Nominal loading of the test item in the saturated solution [%] | ||
Meas. Conc. [µg/L] | Meas. Conc. [µg/L] | |
100 | < LOQ | < LOQ |
QC | 35.8 | 34.0 |
Control | < LOQ | < LOQ |
Meas. conc. = measured concentration of the test item, dilution factors taken into account
LOQ = limit of quantification of the analytical method (30 µg test item/L)
QC = quality control (30 µg test item/L)
Table 5: Validity criteria
Validity Criterion | Required | This study |
Increase of the cell growth in the control cultures | Exponentially, > 16-fold corresponding to a specific growth rate of 0.92 day-1 | 244-fold (specific growth rate 1.83 day-1) |
Mean coefficients of variation for section-by-section specific growth rates (days 0-1, 1-2 and 2-3) in the control cultures |
< 35% | 20.00% |
Coefficient of variation of average specific growth rates during the whole test period in replicate control cultures |
< 7% | 0.60% |
Description of key information
In a GLP-compliant limit test according to OECD Guideline 201 and EU Method C.3 with the unicellular freshwater green alga Pseudokirchneriella subcapitata, the 72-hour NOEL (growth rate) of the test item was determined to be 100 mg/L in nominal. The 72-hour ErL10 and ErL50 were >100 mg/L in nominal (reference 6.1.5-1).
Key value for chemical safety assessment
Additional information
Key information
The toxicity of the test item to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined in a limit-test according to the principles of OECD 201 and Council Regulation (EC) No. 266/2016/Method C.3. The study was conducted over a period of 72 hours under static conditions with an initial cell density of 5416 cells/mL. A saturated solution with a nominal loading rate of 100 mg test item/L was prepared once at room temperature 24 ± 1 hour prior to the start of the exposure. The saturated solution was stirred for 24 ± 1 hours (1100 rpm, room temperature) with a magnetic stirrer. Undissolved particles were removed by membrane filtration. The saturated solution was checked via laser beam (Tyndall effect) for undissolved test item. The Tyndall effect was negative and no undissolved test material was hence detected. Six replicates were tested for the limit loading and six for the control. The test media were clear throughout the test period. The environmental conditions were within the acceptable limits and all validity criteria of the test guidelines were fulfilled. The limit loading rate and the control were analytically verified by HPLC-DAD at the start and the end of the exposure. The measured concentrations of the saturated solution of the test item in the fresh and in the old media were below the Limit of Quantity (LOQ). All effect values given are thus based on the nominal loading of the test item. In result, the 72-hour NOEL for the growth rate was determined to be 100 mg/L. The 72-hour ErL10 and ErL50 were >100 mg/L. The 72-hour NOEL (yield) and EyL10 (yield) were found to be below 100 mg/L. The 72-hour EyL50 (yield) was determined to be >100 mg/L (reference 6.1.5 -1).
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