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EC number: 257-827-7 | CAS number: 52301-70-9
- 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:
- April 14th 27th to 18th, 2020
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Version / remarks:
- 2011
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- Council Regulation (EC) No. 266/2016
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- All concentration levels and the control were analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (72 hours) with algae.
At the start, samples were taken from one additional replicate of each test item concentration and the control and at the end samples were taken from pooled replicates. - Vehicle:
- no
- Details on test solutions:
- Preparation of stock solution: 126 mg test item/l (corresponding to 100 mg a.s./l) were freshly prepared with dilution water. The dispersion was agitated until the solution was visually clear. The stock solution was checked via laser beam (Tyndall effect) for undissolved test item. The Tyndall effect was negative. Preparation of test item solutions: One limit concentration was tested with a nominal concentration of 126 mg test item/l.
- Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Synonyms Selenastrum capricornutum; Ankistrodesmus subcapitata; Raphidocelis subcapitata; Ankistrodesmus bibraianus (Experimental Phycology and Culture Collection of Algae at the University of Goettingen 2014)
- Strain: HINDÁK CCAP 278/4 (axenic)
- Source (laboratory, culture collection): Culture Collection of Algae and Protozoa (CCAP), SAMS Research Services Ltd, Dunstaffnage Marine Laboratory Dunbeg, OBAN; Argyll PA37 1QA; Scotland, UK
- Cultivation at test facility: Fresh stocks are prepared every month on Z-Agar. Light intensity amounts 2567 – 5130 lux for 24 hours per day.
- Culture medium: Nutrient medium Z according to LÜTTGE et al. (1994) - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 72 h
- Hardness:
- 34 mg CaCO3/L
- Test temperature:
- min: 21.0 °C; max: 23 °C; mean value: 22.0 °C
- pH:
- 7.92-8.90 (control)
- Conductivity:
- 136 μS/cm
- Nominal and measured concentrations:
- 126 mg/l (nominal)
- Details on test conditions:
- TEST SYSTEM
- Test vessel: Sterile glass Erlenmeyer flasks, volume: 250 mL, sealed with cotton wool plugs.
- Type: closed
- Fill volume: 100 ml
- Agitation: Test containers were placed on a rotary shaker and oscillated at approximately 70 rpm.
- Initial cells density: Nominal: approximately 5 x 103 - 104 cells/mL; Actual: 5416 cells/mL
- Control end cells density: 1420729 cells/ml (mean, 72hrs)
- No. of vessels per concentration (replicates): six
- No. of vessels per control (replicates): six
- Application: carried out by adding appropriate volumes of the algae inoculum to the test item solutions.
- Preculture: a three days old preculture, prepared in dilution water, was used as inoculum.
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERSv
- Source/preparation of dilution water: OECD TG 201 medium. Ultrapure water was used to prepare the dilution water (conductivity max. 0.1 μS/cm). This medium has a nominal hardness of 0.24 mmol Ca+Mg/L.Hardness, conductivity, TOC, acidity and alkalinity are determined
- Total organic carbon: < LOQ (2.00 mg C/l)
- Acidity: 0.6 mmol/l
- Alkalinity: 0.8 mmol/l
- Intervals of water quality measurement: The pH-values at the start of exposure were measured in one additional replicate of the limit concentration and the control. At the end of exposure, the pH-values were measured from pooled samples of the limit concentration and the control. The room temperature was measured continuously. Light intensity was measured prior to the start of exposure.
OTHER TEST CONDITIONS
- Sterile test conditions: yes; sterile glass flasks were used
- Adjustment of pH: no
- Photoperiod: 24 hours/day light
- Light intensity (target): approximately 4440 to 8880 lux, corresponding to 60 to 120 μE*m-2*s-1
- Light homogeneity (target): within ±15 % over incubation area
EFFECT PARAMETERS MEASURED
- 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 concentration of 126 mg test item/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
- Range finding study: A non-GLP preliminary range finding test under static conditions over a period of 72 hours was conducted at the test facility. A stock solution with a concentration of 126 mg test item/l was freshly prepared with dilution water and agitated until the solution was visually clear. Out of this stock solution two concentration levels were prepared (by dilution) with the concentrations 1.26 and 12.6 mg/l. The stability was assessed in an additional stability test with the Limit test concentration under test conditions with algae. All concentration levels were clear throughout the exposure period
- Results used to determine the conditions for the definitive study: The dilution levels are based on the results of a preliminary range finding test (non-GLP). Growth rate inhibition was 0 % in all tested concentrations. - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate (SIGMA ALDRICH, batch number MKBV0900V, purity 99.0 %, CAS RN 7778-50-9)
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 126 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 126 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- Microscopic evaluation of the cells at the start and end of exposure revealed no morphological abnormalities. The measured concentration of the test item remained stable within ± 20 % of the nominal measured concentration over the test interval at 88 % of the nominal concentration of the test item. All effect values are given based on the nominal test item concentrations. An aged sample and a freshly prepared calibration standard were measured via LC-Q-TOF high resolution-MS for confirmation of the analyte identity. No degradation products were found. It is noteworthy that on basis of the measured accurate masses, only the molecular formula (C40H40N12O22S6) was predicted in the LC-Q-TOF. The chemical structure of the compounds has not been determined. Besides these two isomers, no significant metabolites have been identified by comparison of freshly prepared and aged samples. In comparison to control samples, no additional peaks were visible. The environmental conditions (pH-value, room temperature, light intensity) were determined to be within the acceptable limits in the definitive study. The test media were clear throughout exposure period (possible turbidity related to algae growth not taken into account).
- Results with reference substance (positive control):
- ErC50 (72 hrs) = 0.940 mg/l; 95 % CL: 0.732-1.33 mg/l
Valid range: 0.758 ± 0.535
EyC50 (72 hrs) = 0.660 mg/l; 95 % CL: 0.393-0.700 mg/l
Valid range: 0.414 ± 0.335 - Reported statistics and error estimates:
- The program states no significant difference for the limit concentration compared to the control for growth rate inhibition.
- Validity criteria fulfilled:
- yes
- Remarks:
- Increase of cell growth in control cultures: exponentially ≥16-fold; Mean CVs for section-by-section specific growth rates(days 0-1,1-2,2-3) in ctrl. cultures ≤35 %; CV of average specific growth rates during whole test period in replicate ctrl. cultures
- Conclusions:
- ErC50 (72h) > 126 mg/L (nominal)
NOEC (72h, growth rate) = 126 mg/l (nominal) - Executive summary:
The toxicity of the test item to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 and Council Regulation (EC) No. 266/2016/Method C.3. The aim of the study was the determination of the effects of growth rate and yield over a period of 72 hours in a limit test. The study was conducted under static conditions with an initial cell density of 5416 cells/ml. A stock solution with a nominal test item concentration of 126 mg/l (corresponding to 100 mg a.s./l) was freshly prepared with dilution water. The dispersion was agitated until the solution was visually clear. The stock solution was checked via laser beam (Tyndall effect) for undissolved test item (formation of an emulsion). The Tyndall effect was negative. Six replicates were tested for the limit concentration and six for the control. The test media were clear throughout the test period. The environmental conditions were within the acceptable limits.
The concentration of the test item was analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (72 hours) in the limit concentration and in the control.The measured concentration of the test item was 96 % at the start of the exposure (0 hours) and 88 % at the end of the exposure (72 hours) of the nominal concentration of the test item during the definitive test.
The test item was found not to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours for growth rate. Effect values were determined as follows (based on nominal test item concentration): The EC50-values for both inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were > 126 mg/l. The NOEC-values for both inhibition of growth rate and yield after 72 hours were 126 mg/l.
Reference
Cell Densities
nominal test item concentration (mg/l) | Replicate no. | Cell density [cells/ml] | |||
0 hrs | 24 hrs | 48 hrs | 72 hrs | ||
126 | 1 | 5416 | 26124 | 221039 | 1369712 |
2 | 5416 | 26387 | 213883 | 1463616 | |
3 | 5416 | 24292 | 202366 | 145973 | |
4 | 5416 | 30420 | 212384 | 1345421 | |
5 | 5416 | 27485 | 229140 | 1673670 | |
6 | 5416 | 24628 | 196952 | 1260279 | |
Mean | 5416 | 26556 | 212627 | 1428112 | |
Control | 1 | 5416 | 31233 | 240219 | 1579717 |
2 | 5416 | 28799 | 217846 | 1477928 | |
3 | 5416 | 26086 | 213474 | 1331888 | |
4 | 5416 | 30839 | 228672 | 1277074 | |
5 | 5416 | 29454 | 229918 | 1436111 | |
6 | 5416 | 27351 | 222120 | 1421654 | |
Mean | 5416 | 28960 | 225375 | 1420729 |
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 concentration (mg/l) | Replicate no. | Growth rate (d-1) | Inhibition of Growth rate (%) | Yiels (cells/ml) | Inhibition of yield (%) |
126 | 1 | 1.84 | 1 | 1364296 | 4 |
2 | 1.87 | -1 | 1458200 | -3 | |
3 | 1.87 | 0 | 1450557 | -2 | |
4 | 1.84 | 1 | 1340005 | 5 | |
5 | 1.91 | -3 | 1668254 | -18 | |
6 | 1.82 | 2 | 1254863 | 11 | |
Mean | (-) 1.86 | 0 | (-) 1422696 | -1 | |
Control | 1 | 1.89 | 1574301 | ||
2 | 1.87 | 1472512 | |||
3 | 1.84 | 1326472 | |||
4 | 1.82 | 1271658 | |||
5 | 1.86 | 1430695 | |||
6 | 1.86 | 1416238 | |||
Mean | 1.86 | 1415313 |
Negative values = growth stimulation
Section-by-Section and Average Specific Growth Rates of the Control Group (0 - 72 hours)
Replicate no | Specific growth rate (d-1) section-by-section | Mean (0-72 hrs) | SD ± | CV (%) | Mean CV (%) | |||
0-24 hrs | 24-48 hrs | 48-72 hrs | ||||||
Control | 1 | 1.75 | 2.04 | 1.88 | 1.89 | 0.144 | 7.62 | 10.5 |
2 | 1.67 | 2.02 | 1.92 | 1.87 | 0.181 | 9.65 | ||
3 | 1.57 | 2.10 | 1.83 | 1.84 | 0.265 | 14.4 | ||
4 | 1.74 | 2.00 | 1.72 | 1.82 | 0.158 | 8.70 | ||
5 | 1.69 | 2.06 | 1.83 | 1.86 | 0.182 | 9.80 | ||
6 | 1.62 | 2.09 | 1.86 | 1.86 | 0.238 | 12.8 | ||
Mean | 1.86 | |||||||
SD ± | 0.10 | |||||||
CV (%) | 5.51 |
Environmental Conditions
nominal test item concentration (mg/l) | pH values | |
Start 0 hrs | End 72 hrs | |
126 | 7.84 | 8.81 |
Control | 7.92 | 8.90 |
Room temperature (°C): min: 21.0 °C; max: 23 °C; |
||
Light density (lux) n=14 mean value: 5031; range of the measured values: 4517-5339; equalling -10.2 to 7.32 % CV (%):5.26 | ||
Measured Concentrations and Percent of Nominal Concentration of the Test Item conc during the Definitive Test
Sampling date | Fresh media 0 hrs |
Old media 72 hrs |
||
nominal test item concentration (mg/l) | Test item concentration | |||
Meas. conc. [mg/L] |
% | Meas. conc. [mg/L] |
% of nominal | |
126 | 120 | 96 | 111 | 88 |
QC | 0.978 | 96 | 0.941 | 92 |
Control | < LOQ | < LOQ |
Meas. conc. = measured concentration of the test item, dilution factors taken into account
% = percent of the nominal test item concentration
LOQ = limit of quantification (1.00 mg/l of the test item)
QC = Analytical quality control (1.00 mg/l of the test item)
Description of key information
ErC50 (72h) > 126 mg/L (nominal)
NOEC (72h, growth rate) = 126 mg/l (nominal)
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 126 mg/L
- EC10 or NOEC for freshwater algae:
- 126 mg/L
Additional information
The toxicity of the test item to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 and Council Regulation (EC) No. 266/2016/Method C.3. The study was conducted for 72 hours under static conditions with an initial cell density of 5416 cells/ml. A stock solution with a nominal test item concentration of 126 mg/l (corresponding to 100 mg a.s./l) was freshly prepared with dilution water. Six replicates were tested for the limit concentration and six for the control. The test media were clear throughout the test period. The environmental conditions were within the acceptable limits. The concentration of the test item was analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (72 hours) in the limit concentration and in the control.The measured concentration of the test item was 96 % at the start of the exposure (0 hours) and 88 % at the end of the exposure (72 hours) of the nominal concentration of the test item during the definitive test. The test item was found not to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours for growth rate. Effect values were determined as follows (based on nominal test item concentration): The EC50-values for both inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were >126 mg/l. The NOEC-values for both inhibition of growth rate and yield after 72 hours were 126 mg/l.
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