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EC number: 482-150-0 | CAS number: -
- 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
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- The experimental study was conducted between 21 March 2014 and 1 May 2014
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 014
- Report date:
- 2014
Materials and methods
Test guidelineopen allclose all
- 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
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Magenta T-43
- IUPAC Name:
- Magenta T-43
- Test material form:
- other: solid
- Details on test material:
- Identification: Magenta T-43
Physical state/Appearance: Dark red solid
Storage Conditions: Room temperature in the dark
Constituent 1
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- Range-finding Test:
A sample of each test concentration was taken for chemical analysis at 0 and 72 hours in order to determine the stability of the test item under test conditions. All samples were stored frozen prior to analysis.
Definitive Test:
Samples were taken from the control and the 100 mg/L test group from the bulk test preparation at 0 hours and from the pooled replicates at 72 hours for quantitative analysis. All samples were stored frozen prior to analysis. Duplicate samples were taken at 0 and 72 hours and stored frozen for further analysis if necessary.
Test solutions
- Vehicle:
- no
- Details on test solutions:
- Range-finding Tests:
The test concentration to be used in the definitive test was determined by preliminary range-finding tests. The initial range-finding test was conducted by exposing Pseudokirchnerella subcapitata cells to a series of nominal test concentrations of 0.10, 1.0 and 100 mg/L for a period of 72 hours.
The test was conducted in 250 mL glass conical flasks each containing 100 mL of test preparation and plugged with polyurethane foam bungs to reduce evaporation. Two replicate flasks were used for each control and test concentration. The test item was dissolved directly in culture medium.
An amount of test item (50 mg) was dissolved in culture medium with the aid of ultrasonication for approximately 20 minutes and the volume adjusted to 500 mL to give a 100 mg/L stock solution. A series of dilutions was made from this stock solution to give further stock solutions of 10, 1.0 and 0.10 mgL. An aliquot (450 mL) of each of the stock solutions was separately inoculated with algal suspension (4.4 mL) to give the required test concentrations of 0.10, 1.0, 10 and 100 mg/L.
The stock solutions and each of the prepared concentrations were inverted several times to ensure adequate mixing and homogeneity.
The control group was maintained under identical conditions but not exposed to the test item.
Given that the test item formed colored test solutions it was considered appropriate to conduct spectrophotometer measurements on the stock solutions in order to determine whether significant adsorption of light at the wavelengths required for photosynthesis occurred (460 and 665 nm).
At the start of the initial range-finding test a sample of each test and control culture was removed and the cell density determined using a Coulter® Multisizer Particle Counter. The flasks were then plugged with polyurethane foam bungs and incubated (INFORS Multitron® Version 2 incubator) at 24 ± 1 °C under continuous illumination (intensity approximately 7000 lux) provided by warm white lighting (380-730 nm) and constantly shaken at approximately 150 rpm for 72 hours.
After 72 hours the cell density of each flask was determined using a Coulter® Multisizer Particle Counter.
A sample of each test concentration was taken for chemical analysis at 0 and 72 hours in order to determine the stability of the test item under test conditions. All samples were stored frozen prior to analysis.
The results of the initial range-finding test showed that significant inhibition of growth occured at 100 mg/L where significant light absorbance at 460 nm had been observed. It was therefore considered appropriate to conduct a second range-finding test following the modifications recommended in the OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures with regards to colored test substances.
The test was conducted in 250 mL glass conical flasks each containing 25 mL of test preparation and plugged with polyurethane foam bungs to reduce evaporation. Two replicate flasks were used for each control and test concentration. The test item was dissolved directly in culture medium.
A nominal amount of test item (50 mg) was dissolved in culture medium with the aid of ultrasonication for approximately 1 minute and the volume adjusted to 500 mL to give a 100 mg/L stock solution. A series of dilutions was made from this stock solution to give further stock solutions of 10 and 1.0 mg/L. An aliquot (450 mL) of each of the stock solutions was separately innoculated with algal suspension (3.8 mL) to give the required test concentrations of 1.0, 10 and 100 mg/L.
The stock solutions and each of the prepared concentrations were inversted several times to ensure adequate mixing and homogeneity.
The control group was maintained under identical conditions but not exposed to the test item.
At the start of the second range-finding test a sample of each test and control culture was removed and the cell density determined using a Coulter® Multisizer Particle Counter. The flasks were then plugged with polyurethane foam bungs and incubated (INFORS Multitron® Version 2 incubator) at 24 ± 1°C under continuous illumination (intensity approximately 10000 lux) provided by warm white lighting (380-730 nm) and constantly shaken at approximately 150 rpm for 72 hours.
After 72 hours the cell density of each flask was determine using a Coulter® Multisizer Particle Counter.
Definitive Tests
Based on the result of the second range-finding test a "limit-test" was conducted at a concentration of 100 mg/L under modified test conditions to confirm that no effect on algal growth was observed.
Experimental Preparation
An amount of test item (50 mg) was dissolved in culture medium with the aid of ultrasonication for approximately 3 minutes and the volume adjustedto 500 mL to give a 100 mg/L stock solution. This stock solution was inoculated with algal suspension (3.0 mL) to give the required test concentration of 100 mg/L.
The stock solution and the prepared concentration were inverted several times to ensure adequate mixing and homogeneity.
The concentration and stability of the test item in the test preparations were verified by chemical analysis at 0 and 72 hours.
Test organisms
- Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- The test was carried out using Pseudokirchneriella subcapitata strain CCAP 278/4. Liquid cultures of Pseudokirchneriella subcapitata were obtained from the Culture Collection of Algae and Protozoa (CCAP), SAMS Research Services Ltd, Scottish Marine Institute, Oban, Argyll, Scotland. Master cultures were maintained in the laboratory by the periodic replenishment of culture medium. The master cultures were maintained in the laboratory under
constant aeration and constant illumination at 21 ± 1 °C.
Prior to the start of the test sufficient master culture was added to approximately 100 mL volumes of culture media contained in conical flasks to give an initial cell density of approximately 1 E03 cells/mL. The flasks were plugged with polyurethane foam stoppers and kept under constant agitation by orbital shaker (100 - 150 rpm) and constant illumination at 24 ± 1 °C until the algal cell density was approximately 1 E04 - 1 E05 cells/mL.
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 72 h
Test conditions
- Test temperature:
- 24 ± 1 °C
- pH:
- Control; 0h; pH 7.9
100 mg/L; 0h; pH 8.0
Control;72 h; pH 7.7
100 mg/L; 72 h; pH 7.9 - Nominal and measured concentrations:
- Range-finding test: 0.1, 1.0, 10, 100 mg/L (nominal)
Definitive test: 100 mg/L (nominal); 97 to 104 mg/L (measured) - Details on test conditions:
- Definitive Test:
Exposure Conditions
As in the range-finding tests 250 mL glass conical flasks were used. Six flasks each containing 25 mL of test preparation were used for the control and 100 mg/L treatment group.
The control group was maintained under identical conditions but not exposed to the test item.
Pre-culture conditions gave an algal suspension in log phase growth characterized by a cell density of 8.42 E05 cells per mL. Inoculation of 500 mL of test medium with 3.0 mL of this algal suspension gave an initial nominal cell density of 5 E03 cells per mL and had no significant dilution effect on the final test concentration.
The flasks were plugged with polyurethane foam bungs and incubated (INFORS Multitron® Version 2 incubator) at 24 ± 1 °C under continuous illumination (intensity approximately 10000 lux) provided by warm white lighting (380 - 730 nm) and constantly shaken at approximately 150 rpm for 72 hours.
Evaluations
Test Organism Observations
Samples were taken at 0, 24, 48 and 72 hours and the cell densities determined using a Coulter® Multisizer Particle Counter. To determine the potential effect of the test item on the appearance of algal cells, a sample was removed from each test and control culture (replicates pooled) at the end of the test. The shape and size of the algal cells was inspected microscopically and any abnormalities recorded.
Water Quality Criteria
The pH of the control and 100 mg/L test preparation was determined at initiation of the test and after 72 hours exposure. The pH was measured using a Hach HQ30d Flexi handheld meter. The temperature within the incubator was recorded daily. - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: growth rate and yield
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 100 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: growth rate and yield
- Details on results:
- Range Finding Test:
The cell densities and percentage inhibition of growth values from the exposure of Pseudokirchneriella subcapitata to the test item during the range-finding tests are given in Table 1 and Table 2.
The results of the initial range-finding test performed using standard conditions showed no effect on growth at the test concentrations of 0.10, 1.0 and 10 mg/L. However, growth was observed to be reduced at 100 mg/L. The results of the second range-finding test showed no effect on growth at any of the test concentrations employed.
During the initial range-finding test, spectrophotometer measurements taken at the wavelengths required for photosynthetic growth (460 and 665 nm) showed that the most significant adsorption occurred at 460 nm at 100 mg/L (see table 3). The use of a modified algal inhibition test with increased light intensity and decreased test sample volume overcame this adsorption effect resulting in no significant effect on the algal cells present.
Based on this information a single test concentration of six replicates, of 100 mg/L under modified test conditions was selected for the definitive test. This experimental design conforms to a "limit test" to confirm that at the maximum test concentration given in the OECD/EC Test Guidelines, no effect on growth was observed.
Chemical analysis ofthe 10 and 100 mg/L test preparations taken from the initial range-finding test at 0 and 72 hours showed measured test concentrations torange from 96% to 98% of nominal indicating that the test item was stable under test conditions.
Definitive Test:
Verification of Test Concentrations:
Analysis of the test preparations at 0 and 72 hours showed measured test concentrations to range from 97 to 104 mg/L and so the results are based on nominal test concentrations only.
Growth Data:
Cell density values determined at each sampling time and pH values at 0 and 72 hours are given in Table 4. Daily specific growth rates for the control cultures are given in Table 5. Growth rate and yield values for the control and test cultures after 72 hours and percentage inhibition values are given in Table 6.
From the data given in Table 4 and Table 6, it is clear that the growth rate (r) and yield (y) of Pseudokirchneriella subcapitata (CCAP 278/4) were not affected by the presence of the test item at a nominal test concentration of 100 mg/L over the 72-Hour exposure period. It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.
Accordingly the following results were determined from the data:
Inhibition of Growth Rate:
ErC10 (0 - 72 h) : >100 mg/L
ErC20 (0 - 72 h) :>100 mg/L
ErC50 (0 - 72 h) :>100 mg/L
where ErCx is the test concentration that reduced growth rate by x%.
Statistical analysis of the growth rate data was carried out for the control and 100 mg/L test group using a Student's t-test incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981). There were no statistically significant differences (P≥0.05), between the control and 100 mg/L test group and therefore the "No Observed Effect Concentration" (NOEC) based on growth rate was 100 mg/L.
Inhibition of Yield:
EyC10 (0 - 72 h) : >100 mg/L
EyC20 (0 - 72 h) :>100 mg/L
EyC50 (0 - 72 h) :>100 mg/L
Where:
EyCx is the test concentration that reduced yield by x%.
There were no statistically significant differences (P≥0.05), between the control and 100 mg/L test group and therefore the "No Observed Effect Concentration" (NOEC) based on yield was 100 mg/L. - Results with reference substance (positive control):
- A positive control used potassium dichromate as the reference item at concentrations of 0.25, 0.50, 1.0, 2.0 and 4.0 mg/L.
Exposure conditions and data evaluation for the positive control were similar to those in the definitive test.
Exposure of Pseudokirchneriella subcapitata (CCAP 278/4) to the reference item gave the following results:
ErC50 (0 - 72 h): 1.1 mg/L; 95% confidence limits 0.91 -1.2 mg/L
EyC50 (0 - 72 h): 0.51 mg/L; 95% confidence limits 0.45 - 0.59 mg/L
No Observed Effect Concentration (NOEC) based on growth rate: 0.25 mg/L
No Observed Effect Concentration (NOEC) based on yield: 0.25 mg/L
Lowest Observed Effect Concentration (LOEC) based on growth rate: 0.50 mg/L
Lowest Observed Effect Concentration (LOEC) based on yield: 0.50 mg/L
The results from the positive control with potassium dichromate were within the normal ranges for this reference item. - Reported statistics and error estimates:
- Statistical analysis of the growth rate and yield data was carried out for the control and 100 mg/L test group using a Student's t-test incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981).
Any other information on results incl. tables
Validation Criteria
The following data show that the cell concentration of the control cultures increased by a factor of 110 after 72 hours. This increase was in line with the OECD Guideline that states the enhancement must be at least by a factor of 16 after 72 hours.
Mean cell density of control at 0 hours: 3.68 E03 cells per mL
Mean cell density of control at 72 hours: 4.05 E05 cells per mL
The mean coefficient of variation for section by section specific growth rate for the control cultures was 26% and hence satisfied the validation criterion given in the OECD Guideline which states the mean must not exceed 35%.
The coefficient of variation for average specific growth rate for the control cultures over the test period (0 - 72 h) was 2% and hence satisfied the validation criterion given in the OECD Guideline which states that this must not exceed 7%.
Observations on Cultures
All test and control cultures were inspected microscopically at 72 hours. There were no abnormalities detected in any of the control or test cultures.
Water Quality Criteria
The pH values of the control and 100 mg/L test preparation are given in Table 4. Temperature was maintained at 24 ± 1 °C throughout the test.
The pH value of the control cultures (see Table 4) was observed to range from pH 7.9 at 0 hours to pH 7.7 at 72 hours. The pH deviation in the control cultures was less than 1.5 pH units after 72 hours and therefore was within the limits given in the Test Guidelines.
Observations on Test Item Solubility
At the start of the test all control cultures were observed to be clear colorless solutions whilst the 100 mg/L test cultures were observed to be bright pink solutions. After the 72-Hour test period all control cultures were observed to be green dispersions whilst the 100 mg/L test cultures were observed to be bright pink solutions.
Table 1: Cell Densities and Percentage Inhibition of Growth from the Initial Range-finding Test
Nominal Concentration (mg/L) |
Cell Densities* (cells per mL) |
Inhibition Values (%) |
||
0 Hours |
72 Hours |
Growth Rate |
Yield |
|
Control R1 R2 Mean |
2.75E+03 |
4.79E+05 |
- |
- |
2.83E+03 |
9.30E+05 |
|||
2.79E+03 |
7.05E+05 |
|||
0.10 R1 R2 Mean |
3.15E+03 |
7.87E+05 |
0 |
[21] |
3.38E+03 |
9.20E+05 |
|||
3.26E+03 |
8.53E+05 |
|||
1.0 R1 R2 Mean |
2.26E+03 |
9.30E+05 |
[90 |
[52] |
2.94E+03 |
1.20E+06 |
|||
2.60E+03 |
1.07E+06 |
|||
10 R1 R2 Mean |
2.26E+03 |
8.28E+05 |
0 |
6 |
2.88E+03 |
5.01E+05 |
|||
2.57E+03 |
6.64E+05 |
|||
100 R1 R2 Mean |
2.64E+03 |
1.63E+05 |
25 |
75 |
2.69E+03 |
1.88E+05 |
|||
2.66E+03 |
1.75E+05 |
*Cell densities represent the mean number of cells per mL calculated from the mean of the cell counts from 3 counts for each of the replicate flasks.
R1 and R2 = Replicates 1 an 2
[Increase in growth compared to controls]
Table 2: Cell Densities and Percentage Inhibition of Growth from the Second Range-finding Test
Nominal Concentration (mg/L) |
Cell Densities* (cells per mL) |
Inhibition Values (%) |
||
0 Hours |
72 Hours |
Growth Rate |
Yield |
|
Control R1 R2 Mean |
6.34E+03 |
1.12E+06 |
- |
- |
6.83E+03 |
6.67E+05 |
|||
6.59E+03 |
8.93E+05 |
|||
1.0 R1 R2 Mean |
7.04E+03 |
7.99E+06 |
[3] |
[13] |
5.81E+03 |
1.22E+06 |
|||
6.42E+03 |
1.01E+06 |
|||
10 R1 R2 Mean |
5.54E+03 |
8.14E+05 |
[6] |
[13] |
6.16E+03 |
1.20E+06 |
|||
5.85E+03 |
1.01E+06 |
|||
100 R1 R2 Mean |
5.05E+03 |
1.03E+06 |
[6] |
[16] |
6.16E+03 |
1.04E+06 |
|||
5.60E+03 |
1.03E+06 |
*Cell densities represent the mean number of cells per mL calculated from the mean of the cell counts from 3 counts for each of the replicate flasks.
R1 and R2 = Replicates 1 an 2
[Increase in growth compared to controls]
Table 3: Spectrophotometer Measurements from the Initial Range-Finding Test
Stock Concentration (mg/L) |
Absorbance |
|
460 nm |
665 nm |
|
0.10 |
0.003 |
0.000 |
1.0 |
0.028 |
-0.001 |
10 |
0.273 |
0.000 |
100 |
2.644 |
0.000 |
Table 4: Cell Densities and pH Values in the Definitive Test
Nominal Concentration (mg/L) |
pH |
Cell Densities* (cells per mL) |
pH |
|||
0h |
0h |
24h |
48h |
72h |
72h |
|
Control R1 R2 R3 R4 R5 R6 Mean |
7.9 |
3.81E+03 |
2.04E+04 |
1.33E+05 |
3.95E+05 |
7.7 |
3.81E+03 |
1.93E+04 |
1.34E+05 |
4.05E+05 |
|||
3.61E+03 |
1.86E+04 |
1.37E+05 |
4.55E+05 |
|||
3.05E+03 |
2.14E+04 |
1.39E+05 |
4.38E+05 |
|||
4.17E+03 |
2.17E+04 |
1.16E+05 |
3.40E+05 |
|||
3.64E+03 |
2.10E+04 |
1.28E+05 |
3.98E+05 |
|||
3.68E+03 |
2.04E+04 |
1.31E+05 |
4.05E+05 |
|||
100 R1 R2 R3 R4 R5 R6 Mean |
8.0 |
4.28E+03 |
2.52E+04 |
1.60E+05 |
5.65E+05 |
7.9 |
3.99E+03 |
2.18E+04 |
1.45E+05 |
4.91E+05 |
|||
4.99E+03 |
2.27E+04 |
1.36E+05 |
3.92E+05 |
|||
4.90E+03 |
2.23E+04 |
1.24E+05 |
4.23E+05 |
|||
4.46E+03 |
2.25E+04 |
1.56E+05 |
4.74E+05 |
|||
4.40E+03 |
2.53E+04 |
1.62E+05 |
5.15E+05 |
|||
4.50E+03 |
2.33E+04 |
1.47E+05 |
4.77E+05 |
Table 5: Daily Specific Growth Rates for the Control Cultures in the Definitive Test
|
Daily Specific Growth Rate (cells/mL/hour) |
||
|
Day 0-1 |
Day 1-2 |
Day 2-3 |
Control R1 |
0.059 |
0.078 |
0.045 |
R2 |
0.056 |
0.081 |
0.046 |
R3 |
0.055 |
0.083 |
0.050 |
R4 |
0.061 |
0.078 |
0.048 |
R5 |
0.061 |
0.070 |
0.045 |
R6 |
0.060 |
0.075 |
0.047 |
Mean |
0.059 |
0.078 |
0.047 |
R1-R6 = Replicates
Table 6: Inhibition of Growth Rate and Yield in the Definitive Test
Nominal Concentration (mg/L) |
Growth Rate (cells/mL/hour) |
Yield (cells/mL) |
||
|
0-72 h |
% inhibition |
0-72 h |
% Inhibition |
Control R1 |
0.061 |
|
3.91E+05 |
|
R2 |
0.061 |
|
4.01E+05 |
|
R3 |
0.063 |
|
4.51E+05 |
|
R4 |
0.062 |
|
4.35E+05 |
|
R5 |
0.059 |
|
3.36E+05 |
|
R6 |
0.061 |
|
3.94E+05 |
|
Mean |
0.061 |
|
4.01E+05 |
|
SD |
0.001 |
|
4.02E+04 |
|
100 R1 |
0.066 |
[8] |
5.61E+05 |
|
R2 |
0.064 |
[5] |
4.87E+05 |
|
R3 |
0.061 |
0 |
3.87E+05 |
|
R4 |
0.062 |
[2] |
4.19E+05 |
|
R5 |
0.063 |
[3] |
4.70E+05 |
|
R6 |
0.064 |
[5] |
5.11E+05 |
|
Mean |
0.063 |
[4] |
4.72E+05 |
[18] |
SD |
0.002 |
|
6.28E+04 |
|
*In accordance with the OECD test guideline only the mean value for yield is calculated
R1-R6 = Replicates 1 to 6
SD = Standard Deviation
[Increase in growth as compared to controls]
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Conclusions:
- The effect of the test item on the growth of Pseudokirchneriella subcapitata has been investigated and gave EC50 values of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.
- Executive summary:
Introduction
A study was performed to assess the effect of the test item on the growth of the green alga Pseudokirchneriella subcapitata. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (2006) No 201, "Freshwater Alga and Cyanobacteria, Growth Inhibition Test" referenced as Method C.3 of Commission Regulation (EC) No 761/2009.
Methods
Following preliminary range-finding tests, Pseudokirchneriella subcapitata was exposed to an aqueous solution of the test item at a concentration of 100 mg/L (six replicate flasks) for 72 hours, under constant illumination and shaking at a temperature of 24 ± 1 °C.
Samples of the algal populations were removed daily and cell concentrations determined for each control and treatment group, using a Coulter® Multisizer Particle Counter.
Results
Analysis of the test preparations at 0 and 72 hours showed measured test concentrations to range from 97% to 104% of nominal and so the results are based on nominal test concentrations only.
Exposure of Pseudokirchneriella subcapitata to the test item gave EC50 values of greater than 100 mg/L. The No Observed Effect Concentration was 100 mg/L.
It was considered unnecessary and unrealistic to test at concentrations in excess of 100 mg/L.
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