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EC number: 931-714-9 | CAS number: 35255-57-3
- 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:
- 2016-02-23 to 2016-02-26
- 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:
- (2006)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- (2008)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: 6.25 - 12.50 - 25.0 - 50.0 - 100 mg/L
- Sampling method: The concentrations of the test item were analytically verified from all test concentrations and the control after 0 hours (test start) and 72 hours (test end). Samples at test start were taken from an additional replicate without algae. At end of the test samples were taken directly from the pooled test replicates.
- Sample storage conditions before analysis:All samples were stored at room temperature before preparation and before analysis, if necessary. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Stock solution: A stock solution of 100 mg/L was freshly prepared with dilution water.
Media preparation: direct addition of the test item and shaking until the test item was completely dissolved.
Test concentrations: 5 concentrations were tested in a geometrical series with a dilution factor of 2: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L. - Test organisms (species):
- Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Pseudokirchneriella subcapitata
- Strain: HINDÁK, SAG 61.81
- Source (laboratory, culture collection): SAG Pflanzenphysiologisches Institut der Universitaet Goettingen, Nikolausberger Weg 18, D-37073 Goettingen, Germany
- Age of inoculum (at test initiation): 4 days
- Method of cultivation: Fresh stocks are prepared every month on Z-Agar. Light intensity amounted to 35-70 µE·m-2·s-1 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:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- No
- Hardness:
- 0.24 mmol Ca+Mg/l
- Test temperature:
- Nominal range: 21 - 24 °C, controlled at ± 2 °C
- Dissolved oxygen:
- Not measured
- Salinity:
- Not measured, freshwater conditions
- Nominal and measured concentrations:
- Nominal: 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L.
- Details on test conditions:
- TEST SYSTEM
- Test vessel: 250 mL Erlenmeyer flasks
- Type (delete if not applicable): open, cotton wool plugs
- Material, size, headspace, fill volume: sterile 250 mL Erlenmeyer flasks, test volume 100 mL
- Aeration: Test containers were placed on a rotary shaker and oscillated at appr. 70 rpm
- Type of flow-through (e.g. peristaltic or proportional diluter): None
- Renewal rate of test solution (frequency/flow rate): One application at test start
- Initial cells density: Nominal: approximately 5 x E03 - E04 cells/mL, Actual: mean 6305 cells/mL
- Control end cells density: Mean 1592351 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:
Composition of Dilution Water
according to the guideline
Component Concentration [mg/L]
NH4Cl 15
MgCl2 6 H2O 12
CaCl2 2 H2O 18
MgSO4 7 H2O 15
KH2PO4 1.6
FeCl3 6 H2O 0.064
Na2EDTA 2 H2O 0.1
H3BO3 0.185
MnCl2 4 H2O 0.415
ZnCl2 3 x 10-3
Na2MoO4 2 H2O 7 x 10-3
CoCl2 6 H2O 1.5 x 10-3
CuCl2 2 H2O 1 x 10-5
NaHCO3 50
pH 8.1 0.2
OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: no
- Photoperiod: 24 h
- Light intensity and quality: 60 - 120 µE x m-2 x s-1, mean value: 75.3
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter] fluorimeter, Cell density and self fluorescence was measured daily via Chlorophyll-a- fluorescence, excitation at 436 nm, emission at 685 nm. Dilution water was used as background signal.
TEST CONCENTRATIONS
Results of the Range Finding Test (0-72 hours)
Nominal Test Item Concentration [mg/L] Growth Rate Inhibition [%] Yield Inhibition [%]
100 12 45
10.0 -1 -5
1.00 -8 -56
- Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 45.7 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 12.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 25.7 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Details on results:
- - Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test):
- Unusual cell shape: No
- Colour differences: No
- Flocculation: No
- Adherence to test vessels: No
- Aggregation of algal cells: No
- Other: No
- Any observations (e.g. precipitation) 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):
- - Results with reference substance valid? Yes
EC50-Values of the Reference Item based on nominal concentrations [mg/L], (0-72 hours)
Current Study Valid Range (average ± 3 x SD)
Growth Rate inhibition
ErC50 0.564 0.797 ± 0.428
95 % confidence interval 0.546 - 0.582
Yield inhibition
EyC50 0.322 0.415 ± 0.275
95 % confidence interval 0.239 - 0.342
SD = Standard deviation - Reported statistics and error estimates:
- NOEC/LOEC was determined by calculation of statistical significance of growth rate and yield. As standard One Way Analysis of Variance (ANOVA) and DUNNETT’s test were used for NOEC/LOEC calculations. When running a One Way Analysis of Variance a Normality test and an Equal Variance test were done first. The SHAPIRO-WILK-Test was used to test for normally distributed populations. The Levene median test was used for equal variance. P-values for both Normality and Equal Variance tests are 0.05. The -value (acceptable probability of incorrectly concluding that there is a difference) is =0.05. Detection of statistically significant differences was considered negligible when inhibition values were ≤ 10 %. These findings were classified as “biologically not significant”. Following the recommendations of the UBA (HEGER ET AL. 1998), inhibitions > 10 % are significant.
EC10-, EC20- and EC50-values of the growth rate and yield inhibition after 72 hours were calculated by sigmoidal dose-response regression. Calculation of the confidence intervals
EC10-, EC20- and EC50-values were carried out using standard procedures. - Validity criteria fulfilled:
- yes
- Conclusions:
- In this study, the test substance was found to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours with the following effect values (based on nominal test item concentrations): The EC50-values with 95 % confidence intervals for inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were 45.7 (43.2 – 47.9) mg/L and 26.1 (22.6 – 31.7) mg/L, respectively. The NOEC-values for both inhibition of growth rate and yield after 72 hours were 12.5 mg/L. All effect level are based on nominal concentration which had been analytically verified.
- Executive summary:
The toxicity of the test substance to the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 and Council Regulation (EC) No. 761/2009/Method C.3.The aim of the study was the determination of NOEC, LOEC, EC10-, EC20- and EC50-values of growth rate and yield over a period of 72 hours.
The study was conducted under static conditions with an initial cell density of 6305 cells/mL. A stock solution of 100 mg/L was prepared by direct addition of the test item to the dilution water and shaken until the test item was completely dissolved. Five concentrations were tested in a geometrical series with a dilution factor of 2.0 (nominal): 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L. Three replicates were tested for each test item concentration and six replicates for the control. The environmental conditionswere within the acceptable limits.
The concentrations of the test subsance were determined in the fresh media (0 h) and old media (72 h) of all tested concentration levels and the control via LC-MS/MS.
The measured concentrations in the fresh media (0 h) were in the range of 111 to 118% of the nominal values at tested concentration levels. The measured concentrations in the old media (72 h) were in the range of 106 to 120% of the nominal values of all concentrations. Therefore, all effect values given are based on the nominal test item concentrations. In this study, the test substance was found to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours with the following effect values (based on nominal test item concentrations): The EC50-values with 95 % confidence intervals for inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were 45.7 (43.2 – 47.9) mg/L and 26.1 (22.6 – 31.7) mg/L, respectively. The NOEC-values for both inhibition of growth rate and yield after 72 hours were 12.5 mg/L.
Reference
Cell densities
Nominal test item concentration |
Replicate |
Cell density [cells/mL] |
|||
[mg/L] |
No. |
0 hours |
24 hours |
48 hours |
72 hours |
100 |
1 |
6305 |
5242 |
5620 |
4526 |
2 |
6305 |
5682 |
5902 |
6412 |
|
3 |
6305 |
5327 |
6302 |
4574 |
|
Mean |
6305 |
5417 |
5941 |
5171 |
|
50.0 |
1 |
6305 |
10438 |
23955 |
76118 |
2 |
6305 |
11393 |
25181 |
70370 |
|
3 |
6305 |
10942 |
22875 |
43975 |
|
Mean |
6305 |
10924 |
24004 |
63488 |
|
25.0 |
1 |
6305 |
14260 |
76718 |
715064 |
2 |
6305 |
17215 |
127094 |
1084155 |
|
3 |
6305 |
15565 |
82689 |
838113 |
|
Mean |
6305 |
15680 |
95500 |
879111 |
|
12.5 |
1 |
6305 |
16896 |
142925 |
1092415 |
2 |
6305 |
18937 |
171369 |
1530995 |
|
3 |
6305 |
18263 |
134434 |
1160071 |
|
Mean |
6305 |
18032 |
149576 |
1261160 |
|
6.25 |
1 |
6305 |
17747 |
152092 |
1176703 |
2 |
6305 |
22655 |
204520 |
1560651 |
|
3 |
6305 |
26946 |
234571 |
1637356 |
|
Mean |
6305 |
22449 |
197061 |
1458237 |
|
Control |
1 |
6305 |
28186 |
229091 |
1505934 |
2 |
6305 |
27639 |
245740 |
1751356 |
|
3 |
6305 |
24217 |
205761 |
1567670 |
|
4 |
6305 |
22892 |
164440 |
1353990 |
|
5 |
6305 |
29145 |
248587 |
1594338 |
|
6 |
6305 |
30777 |
224022 |
1780815 |
|
Mean |
6305 |
27143 |
219607 |
1592351 |
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 |
Replicate |
Growth rate |
Inhibition of growth rate |
Yield |
Inhibition of yield |
||
[mg/L] |
No. |
[d-1] |
[%] |
[cells/mL] |
[%] |
||
100 |
1 |
|
-0.111 |
100 |
|
-1779 |
100 |
2 |
|
0.006 |
100 |
|
107 |
100 |
|
3 |
|
-0.107 |
100 |
|
-1731 |
100 |
|
Mean |
(+) |
-0.071 |
100 |
(+) |
-1134 |
100 |
|
50.0 |
1 |
|
0.830 |
55 |
|
69813 |
96 |
2 |
|
0.804 |
56 |
|
64065 |
96 |
|
3 |
|
0.647 |
65 |
|
37670 |
98 |
|
Mean |
(+) |
0.761 |
59 |
(+) |
57183 |
96 |
|
25.0 |
1 |
|
1.58 |
14 |
|
708759 |
55 |
2 |
|
1.72 |
7 |
|
1077850 |
32 |
|
3 |
|
1.63 |
12 |
|
831808 |
48 |
|
Mean |
(+) |
1.64 |
11 |
(+) |
872806 |
45 |
|
12.5 |
1 |
|
1.72 |
7 |
|
1086110 |
32 |
2 |
|
1.83 |
1 |
|
1524690 |
4 |
|
3 |
|
1.74 |
6 |
|
1153766 |
27 |
|
Mean |
(-) |
1.76 |
4 |
(-) |
1254855 |
21 |
|
6.25 |
1 |
|
1.74 |
5 |
|
1170398 |
26 |
2 |
|
1.84 |
0 |
|
1554346 |
2 |
|
3 |
|
1.85 |
-1 |
|
1631051 |
-3 |
|
Mean |
(-) |
1.81 |
2 |
(-) |
1451932 |
8 |
|
Control |
1 |
|
1.83 |
|
|
1499629 |
|
2 |
|
1.88 |
|
|
1745051 |
|
|
3 |
|
1.84 |
|
|
1561365 |
|
|
4 |
|
1.79 |
|
|
1347685 |
|
|
5 |
|
1.84 |
|
|
1588033 |
|
|
6 |
|
1.88 |
|
|
1774510 |
|
|
Mean |
|
1.84 |
|
|
1586046 |
|
Section-by-Section and Average Specific Growth Rates of the Control Group (0 - 72 hours)
|
Replicate No. |
Specific growth rate [d-1] |
Mean (0 - 72 hours) |
SD ± |
CV |
Mean CV [%] |
||
section-by-section |
||||||||
0 - 24 hours |
24 - 48 hours |
48 - 72 hours |
||||||
Control |
1 |
1.50 |
2.10 |
1.88 |
1.83 |
0.303 |
16.6 |
19.0 |
2 |
1.48 |
2.19 |
1.96 |
1.88 |
0.362 |
19.3 |
||
3 |
1.35 |
2.14 |
2.03 |
1.84 |
0.430 |
23.4 |
||
4 |
1.29 |
1.97 |
2.11 |
1.79 |
0.439 |
24.5 |
||
5 |
1.53 |
2.14 |
1.86 |
1.84 |
0.307 |
16.6 |
||
6 |
1.59 |
1.99 |
2.07 |
1.88 |
0.260 |
13.8 |
||
|
|
|
Mean |
1.84 |
|
|
||
|
|
|
SD ± |
0.03 |
|
|||
|
|
|
CV [%] |
1.83 |
|
SD = Standard deviation CV = Coefficient of variation
Description of key information
The toxicity of the test substanceto the unicellular freshwater green alga Pseudokirchneriella subcapitata was determined according to the principles of OECD 201 andCouncil Regulation (EC) No. 761/2009/Method C.3.The aim of the study was the determination of NOEC, LOEC, EC10-, EC20- and EC50-values of growth rate and yield over a period of 72 hours.
The study was conducted under static conditions with an initial cell density of 6305 cells/mL. A stock solution of 100 mg/L was prepared by direct addition of the test item to the dilution water and shaken until the test item was completely dissolved. Five concentrations were tested in a geometrical series with a dilution factor of 2.0 (nominal): 6.25 - 12.5 - 25.0 - 50.0 - 100 mg/L. Three replicates were tested for each test item concentration and six replicates for the control. The environmental conditionswere within the acceptable limits.
The concentrations of the test
subsance were determined in the fresh media (0 h) and old media (72 h)
of all tested concentration levels and the control via LC-MS/MS.
The measured concentrations in the fresh media (0 h) were in the range
of 111 to 118% of the nominal values at tested concentration levels. The
measured concentrations in the old media (72 h) were in the range of 106
to 120% of the nominal values of all concentrations. Therefore, all
effect values given are based on the nominal test item concentrations.In
this study, the test substance was found to inhibit the growth of the
freshwater green alga Pseudokirchneriella subcapitata after 72 hours
with the following effect values (based on nominal test item
concentrations): The EC50-values with 95 % confidence intervals for
inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were
45.7 (43.2 – 47.9) mg/L and 26.1 (22.6 – 31.7) mg/L, respectively. The
NOEC-values for both inhibition of growth rate and yield after 72 hours
were 12.5 mg/L.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 45.7 mg/L
- EC10 or NOEC for freshwater algae:
- 12.5 mg/L
Additional information
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.