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EC number: - | 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
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:
- Experiment start date - 12 March 2010; Experiment completion date - 23 March 2010; Study completion date - 27 May 2010.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- Identification: FAT 40851/A TE
Batch Number: TZ 5891 / BOP 02-09
Purity: 69.9% all coloured components
Appearance: Orange powder
Expiry Date: July 31, 2014
Storage Conditions: At room temperature at about 20 °C - Analytical monitoring:
- yes
- Details on sampling:
- For measurement of the actual concentrations of the test item, duplicate samples were taken from the test media of all test concentrations at the start of the test (without algae) and at the end of the test (containing algae). At the same sampling times, duplicate samples were also taken from the control.
For the 72-hour stability samples, additional flasks containing the test medium with algae were incubated for each treatment under the test conditions. This was necessary as the volume of test solution of the treatment replicates (3 mL) was too small to perform the analyses.
All samples were stored deep-frozen (at about -20 °C) immediately after sampling until analysis. In a pre-test (non-GLP) the samples proved to be stable under these storage conditions. - Vehicle:
- no
- Details on test solutions:
- The test medium of the highest nominal concentration of 100 mg/L was prepared by dissolving 103.76 mg of the test item completely in 1038 mL of test water using ultrasonic treatment for 15 minutes and intense stirring for 15 minutes at room temperature. The test medium of the highest test concentration was diluted with test water to prepare the test media of the lower test concentrations. The test media were prepared just before the start of the test.
- Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) Nygaard et al (1986) recommended describing the taxa within the Genus Raphidocelis HINDAK as: Raphidocelis subcapitata (KORSIKOV) nov. comb.
Basionym: Ankistrodesmus subcapitatus KORSIKOV
Syn.: Kirchneriella subcapitata KORSIKOV
Syn.: Selenastrum capricornutum PRINTZ
Syn.: NIVA-CHL 1
∙ NYGAARD, G., KOMÁREK, J., KRISTIANSEN, J., SKULBERG O.M. (1986). Taxonomic designations of the bioassay alga NIVA-CHL 1 ("Selenastrum capricornutum") and some related strains. Opera Botanica 90:5-46
- Strain: Strain No. 61.81 SAG
- Source: supplied by the Collection of Algal Cultures (SAG, Institute for Plant Physiology, University of Göttingen, 37073 Göttingen / Germany).
- Method of cultivation: The algae were cultivated at Harlan Laboratories under standardized conditions according to the test guidelines.
ACCLIMATION
- Acclimation period: An inoculum culture was set up three days before the start of the exposure. The algae were cultivated under the test conditions and were kept in the exponential growth phase until inoculation of the test solutions.
- Culturing media and conditions: Same as test
- Any deformed or abnormal cells observed: None - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Hardness:
- The water hardness (calculated) of the test water was 0.24 mmol/L (= 24 mg/L as CaCO3).
- Test temperature:
- The test flasks were incubated in a temperature-controlled water bath at a temperature of 22 °C.
- pH:
- Start: 7.9 - 8.0, End: 7.9 - 8.2
- Dissolved oxygen:
- During exposure, the test solutions were continuously stirred by magnetic stirrers.
- Salinity:
- Reconstituted test water prepared according to the test guidelines was used for algal cultivation and testing (see Table M1 below)
- Nominal and measured concentrations:
- 0, 1.0, 3.2, 10, 32 and 100 mg/L nominal
The concentrations of the test item test substance were determined in the duplicate test medium samples from the nominal test concentrations of 3.2 to 100 mg/L. The samples from the nominal test concentration 1.0 mg/L were not analyzed, since this concentrations was below the NOEC determined in this test. From the control samples, one of the duplicate samples was analyzed from the corresponding sampling times.
The measured concentrations of the test item in the test media of the test concentrations of 3.2 to 100 mg/L were between 86 and 106% of the nominal values at the start and the end of the test. Thus, the correct dosing of the test item test substance at test start was confirmed. Furthermore, it has been demonstrated that the test item was stable during the test period of 72 hours under the conditions of the test, and the reported biological results are based on the nominal concentrations of the test item. - Details on test conditions:
- TEST SYSTEM
- Test vessel: 100 mL Erlenmeyer flasks
- Type: Each test flask was covered with a glass dish.
- Material, size, headspace, fill volume: Since the test water was significantly colored by the test item, 17 mL of test solution were placed in 100 mL Erlenmeyer flasks achieving a reduced layer thickness of 0.9 cm and enhancing the light supply for the algae.
Aeration: During exposure, the test solutions were continuously stirred by magnetic stirrers.
- Initial cells density: 10'000 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Sterile purified water
OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: no
- Temperature: The test flasks were incubated in a temperature-controlled water bath at a temperature of 22 °C
- Photoperiod: 24 h / day
- Light intensity and quality: The test flasks were illuminated by fluorescent tubes (Philips TLD 36W-1/840), installed above the test flasks. The test flasks were positioned randomly and repositioned daily. The mean measured light intensity at the level of the test solutions was approximately 7960 Lux (range: 6970 to 8650 Lux, measured at nine places in the experimental area). The light intensity was within ± 15% from the average light intensity as recommended by the guideline. The shadowing effect of each test concentration was determined by measurement of the light intensity under each test solution: In the nominal test item concentration (mg/L) of 0, 1.0, 3.2, 10, 32, and 100 mg/L the Light intensity (Lux) was 8300, 8360, 8230, 7950, 7710, and 6990 respectively.
EFFECT PARAMETERS MEASURED :
- Determination of cell concentrations: The algal biomass in the samples was determined by measurement of the algal cell density using an electronic particle counter (Coulter Counter, Model ZM). The measurements were performed at least in duplicate.
- At the end of the test, a sample was taken from the control and from the test concentration of nominal 100 mg/L to determine a potential influence of the test item on the algal cells. The shape and size of the algal cells were visually inspected.
- The pH was measured and recorded in each treatment at the start and at the end of the test.
- The water temperature was measured and recorded daily in an Erlenmeyer flask filled with water and incubated under the same conditions as the test flasks.
- The appearance of the test media was also recorded daily.
TEST CONCENTRATIONS
- Range finding study: Yes
- Test concentrations: Based on the range-finding study - Reference substance (positive control):
- yes
- Remarks:
- potassium dichromate
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 48 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 10 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 32 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 32 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Details on results:
- - Exponential growth in the control (for algal test): yes
- The microscopic examination of the algal cells at the end of the test showed no difference between the algae growing at the nominal test concentration of 100 mg/L and the algal cells in the control. The shape and size of the algal cells were obviously not affected by the test item up to at least this concentration.
- In the control the biomass increased by a factor of 128 over 72 hours. The validity criterion of increase of biomass by at least a factor of 16 within three days was fulfilled. The mean coefficient of variation of the daily growth rates in the control (section-by-section growth rates) during 72 hours was 16%. According to the OECD test guideline, the mean coefficient of variation must not be higher than 35%. Thus, the validity criterion was fulfilled. The coefficient of variation of the average specific growth rates in the replicates of the control after 72 hours was 1.0%. According to the OECD test guideline, the coefficient of variation must not be higher than 7%. Thus, the validity criterion was fulfilled.
- No remarkable observations were made concerning the appearance of the test media. All test media were clear orange colored solutions throughout the test period.
- At the start of the test, the pH measured in the treatments was between 7.9 and 8.0. At the end of the test, pH values of 7.9 to 8.2 were measured. The water temperature during the test was maintained at 22 °C. - Results with reference substance (positive control):
- - Results with reference substance valid: Yes
- EC50: 0.75 mg/L (72h) (Sept 2009)
For evaluation of the algal quality and experimental conditions, potassium dichromate is tested as a positive control twice a year to demonstrate satisfactory test conditions. The result of the latest positive control test performed in September 2009 showed that the sensitivity of the test system was within the internal historical range (72-hour EC50 for the growth rate: 0.75 mg/L (study C63678), range of the 72-hour EC50 for the growth rate from 2000 to 2009: 0.71 - 1.7 mg/L). - Reported statistics and error estimates:
- The R² fit of the calibration curve used was 1.0000.
The test item had a statistically significant inhibitory effect on the growth of the algae (average growth rate and yield) after the test period of 72 hours at the two highest test concentrations of 32 mg/L and 100 mg/L (results of Williams tests, one-sided, α = 0.05, Table 2 and Table 3). Thus, this concentration was determined to be the 72-hour LOEC.
The 72-hour NOEC was determined to be 10 mg/L, since up to and including this test concentration, the growth rate and yield of the algae after 72 hours were not statistically significantly lower than in the control. - Validity criteria fulfilled:
- yes
- Conclusions:
- The test substance is considered not acute toxic to aquatic freshwater green algae.
- Executive summary:
The influence of the test item test substance on the growth of a freshwater green algal species (Pseudokirchneriella subcapitata) was investigated in a 72-hour static test under GLP according to OECD Guideline 201 (2006), the EU Commission Directive 92/69/EEC, C.3 (1992) and the Commission Regulation (EC) No 440/2008, C.3.
The nominal concentrations of the test item of 1.0, 3.2, 10, 32 and 100 mg/L were tested in parallel with a control. The measured concentrations of the test item in the test media of the test concentrations of 3.2 to 100 mg/L were between 86 and 106% of the nominal values at the start and the end of the test. Thus, the correct dosing of the test item test substance at test start was confirmed. Furthermore, it has been demonstrated that the test item was stable during the test period of 72 hours under the conditions of the test, and the reported biological results are based on the nominal concentrations of the test item.
The test item had a statistically significant inhibitory effect on the growth of the algae (average growth rate and yield) after the test period of 72 hours at the two highest test concentrations of 32 mg/L and 100 mg/L (results of Williams tests, one-sided, α = 0.05). Thus, this concentration was determined to be the 72-hour LOEC. The 72-hour NOEC was determined to be 10 mg/L, since up to and including this test concentration, the growth rate and yield of the algae after 72 hours were not statistically significantly lower than in the control. As not 50% effect level was reached, no EC50 was established.
The microscopic examination of the algal cells at the end of the test showed no difference between the algae growing at the nominal test concentration of 100 mg/L and the algal cells in the control. The shape and size of the algal cells were obviously not affected by the test item up to and including this concentration.
In the control the biomass increased by a factor of 128 over 72 hours. The validity criterion of increase of biomass by at least a factor of 16 within three days was fulfilled. The mean coefficient of variation of the daily growth rates in the control (section-by-section growth rates, during 72 hours was 16%. According to the OECD test guideline, the mean coefficient of variation must not be higher than 35%. Thus, the validity criterion was fulfilled. The coefficient of variation of the average specific growth rates in the replicates of the control after 72 hours was 1.0%. According to the OECD test guideline, the coefficient of variation must not be higher than 7%. Thus, the validity criterion was fulfilled. No remarkable observations were made concerning the appearance of the test media. All test media were clear orange coloured solutions throughout the test period. At the start of the test, the pH measured in the treatments was between 7.9 and 8.0. At the end of the test, pH values of 7.9 to 8.2 were measured. The water temperature during the test was maintained at 22 °C.
In conclusion the test item test substance is considered not acute toxic to aquatic freshwater green algae.
Reference
The tabulated values represent rounded results obtained by calculation using the exact raw data.
Table 1: Biomass of Algae
Nominal test item concentration [mg/L] |
Rep. no. |
Biomass of algae* |
||
24 hours |
48 hours |
72 hours |
||
Control |
1 |
9.0 |
44.0 |
298.9 |
2 |
10.4 |
53.5 |
321.9 |
|
3 |
10.5 |
64.6 |
299.0 |
|
4 |
8.3 |
44.6 |
306.4 |
|
5 |
9.5 |
58.7 |
287.3 |
|
6 |
8.3 |
52.2 |
329.0 |
|
Mean |
9.3 |
52.9 |
307.1 |
|
SD |
1.0 |
8.0 |
15.6 |
|
1.0 |
1 |
9.2 |
53.1 |
245.7 |
2 |
9.5 |
43.8 |
341.1 |
|
3 |
8.2 |
53.4 |
321.4 |
|
Mean |
8.9 |
50.1 |
302.7 |
|
SD |
0.7 |
5.5 |
50.4 |
|
3.2 |
1 |
9.4 |
50.6 |
236.9 |
2 |
9.9 |
53.2 |
299.6 |
|
3 |
8.3 |
59.6 |
289.8 |
|
Mean |
9.2 |
54.5 |
275.4 |
|
SD |
0.8 |
4.6 |
33.7 |
|
10 |
1 |
9.8 |
44.3 |
238.6 |
2 |
7.4 |
41.4 |
354.8 |
|
3 |
7.8 |
48.4 |
287.7 |
|
Mean |
8.4 |
44.7 |
293.7 |
|
SD |
1.3 |
3.5 |
58.3 |
|
32 |
1 |
6.0 |
35.3 |
226.4 |
2 |
5.4 |
33.3 |
312.8 |
|
3 |
6.3 |
33.8 |
233.3 |
|
Mean |
5.9 |
34.1 |
257.5 |
|
SD |
0.5 |
1.1 |
48.0 |
|
100 |
1 |
2.8 |
16.9 |
225.7 |
2 |
2.5 |
16.3 |
234.7 |
|
3 |
2.2 |
16.6 |
227.0 |
|
Mean |
2.5 |
16.6 |
229.1 |
|
SD |
0.3 |
0.3 |
4.9 |
SD: Standard deviation
* The biomass was determined by fluorescence measurement (at least duplicate measurements per replicate) and is given as relative fluorescence units ( ∙ 10³). At the start of the test, the initial cell density was 10’000 algal cells/mL, corresponding to 2.40 x 10³ relative fluorescence units.
Table 2: Average Growth Rates (µ)
Nominal |
Average growth rate µ [day-1] and inhibition of µ (Ir) |
|||||
0-24 h |
0-48 h |
0-72 h |
||||
µ |
Ir[%] |
µ |
Ir[%] |
µ |
Ir[%] |
|
Control |
1.35 |
0.0 |
1.54 |
0.0 |
1.62 |
0.0 |
1.0 |
1.31 |
3.0 |
1.52 |
1.6 |
1.61 |
0.5 |
3.2 |
1.34 |
1.0 |
1.56 |
-1.2 |
1.58 |
2.3 |
10 |
1.24 |
8.3 |
1.46* |
5.2 |
1.60 |
1.2 |
32 |
0.90* |
33.8 |
1.33* |
14.0 |
1.55* |
3.8 |
100 |
0.04* |
97.3 |
0.97* |
37.3 |
1.52* |
6.0 |
*: mean value statistically significantly lower than in the control (according to Williams-test, one-sided, α= 0.05)
---: not analyzed
Table 3: Yield(Y)
Nominal |
Yield Y ( ∙ 10³) and inhibition of Y (Iy) |
|||||
0-24 h |
0-48 h |
0-72 h |
||||
Y |
Iy [%] |
Y |
Iy [%] |
Y |
Iy [%] |
|
Control |
6.9 |
0.0 |
50.5 |
0.0 |
304.7 |
0.0 |
1.0 |
6.5 |
5.7 |
47.7 |
5.6 |
300.3 |
1.4 |
3.2 |
6.8 |
2.0 |
52.1 |
-3.0 |
273.0 |
10.4 |
10 |
6.0 |
14.0 |
42.3* |
16.3 |
291.3 |
4.4 |
32 |
3.5* |
49.6 |
31.7* |
37.2 |
255.1* |
16.3 |
100 |
0.1* |
98.5 |
14.2* |
71.9 |
226.7* |
25.6 |
*: mean value statistically significantly lower than in the solvent control (according to Williams-test, one-sided,α= 0.05)
---: not analyzed
Table 4: Section-by-Section Growth Rates
Nominal [mg/L] |
Section-by-section growth rates [day-1] |
|||||
0-24 h |
24-48 h |
48-72 h |
||||
µ |
Ir [%] |
µ |
Ir [%] |
µ |
Ir [%] |
|
Control |
1.35 |
0.0 |
1.73 |
0.0 |
1.77 |
0.0 |
1.0 |
1.31 |
3.0 |
1.72 |
0.5 |
1.79 |
-1.5 |
3.2 |
1.34 |
1.0 |
1.78 |
-2.8 |
1.62 |
8.4 |
10 |
1.24 |
8.3 |
1.68 |
2.8 |
1.87 |
-5.9 |
32 |
0.90 |
33.8 |
1.76 |
-1.6 |
2.01 |
-13.8 |
100 |
0.04 |
97.3 |
1.90 |
-9.6 |
2.63 |
-48.6 |
---: not analyzed
Table 5: pH Values in theTreatments
Nominal |
pH values |
|
Start |
End |
|
Control |
8.0 |
7.9 |
1.0 |
8.0 |
8.0 |
3.2 |
8.0 |
8.2 |
10 |
7.9 |
8.2 |
32 |
7.9 |
8.2 |
100 |
7.9 |
8.1 |
Description of key information
The EC50 and the NOEC for growth rate were >100 and 10 mg/L, respectively.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 100 mg/L
- EC10 or NOEC for freshwater algae:
- 10 mg/L
Additional information
The influence of the test item test substance on the growth of a freshwater green algal species (Pseudokirchneriella subcapitata) was investigated in a 72-hour static test under GLP according to OECD Guideline 201 (2006), the EU Commission Directive 92/69/EEC, C.3 (1992) and the Commission Regulation (EC) No 440/2008, C.3. The nominal concentrations of the test item of 1.0, 3.2, 10, 32 and 100 mg/L were tested in parallel with a control. The measured concentrations of the test item in the test media of the test concentrations of 3.2 to 100 mg/L were between 86 and 106 % of the nominal values at the start and the end of the test. Thus, the correct dosing of the test item test substance at test start was confirmed. Furthermore, it has been demonstrated that the test item was stable during the test period of 72 hours under the conditions of the test, and the reported biological results are based on the nominal concentrations of the test item. The test item had a statistically significant inhibitory effect on the growth of the algae (average growth rate and yield) after the test period of 72 hours at the two highest test concentrations of 32 mg/L and 100 mg/L (results of Williams tests, one-sided, α = 0.05). Thus, this concentration was determined to be the 72-hour LOEC. The 72-hour NOEC was determined to be 10 mg/L, since up to and including this test concentration, the growth rate and yield of the algae after 72 hours were not statistically significantly lower than in the control. As 50 % effect level was not reached, no EC50 was established. The microscopic examination of the algal cells at the end of the test showed no difference between the algae growing at the nominal test concentration of 100 mg/L and the algal cells in the control. The shape and size of the algal cells were obviously not affected by the test item up to and including this concentration. In the control the biomass increased by a factor of 128 over 72 hours. The validity criterion of increase of biomass by at least a factor of 16 within three days was fulfilled. The mean coefficient of variation of the daily growth rates in the control (section-by-section growth rates, during 72 hours was 16 %. According to the OECD test guideline, the mean coefficient of variation must not be higher than 35 %. Thus, the validity criterion was fulfilled. The coefficient of variation of the average specific growth rates in the replicates of the control after 72 hours was 1.0 %. According to the OECD test guideline, the coefficient of variation must not be higher than 7 %. Thus, the validity criterion was fulfilled. No remarkable observations were made concerning the appearance of the test media. All test media were clear orange coloured solutions throughout the test period. At the start of the test, the pH measured in the treatments was between 7.9 and 8.0. At the end of the test, pH values of 7.9 to 8.2 were measured. The water temperature during the test was maintained at 22 °C. In conclusion the test item test substance is considered not acute toxic to aquatic freshwater green algae.
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