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EC number: 279-242-6 | CAS number: 79720-19-7
- 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:
- 2016-02-01 to 2016-02-19
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- 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
- Analytical monitoring:
- yes
- Details on sampling:
- Determination of the test item
Analytical evaluation of the concentrations of the test item was carried out via LC-MS/MS from freshly prepared media after 0 hours and old test media after 72 hours of exposure. Samples for the test item analysis at the beginning of the exposure were taken from additionally prepared replicates with algae. For the test item analysis after 72 hours, samples were taken from the test replicates. - Vehicle:
- no
- Details on test solutions:
- Preparation of the saturated solution
A dispersion with a nominal loading of 100 mg/L of the test item was prepared with dilution water one day prior to the start of the exposure. An appropriate amount of the test item was weighed out and transferred with an appropriate amount of dilution water into a measuring flask. This dispersion was stirred for 24 hours with approximately 1100 rpm at room temperature. Then, undissolved test item was removed by membrane filtration (membrane filter 0.45 µm, RC, MACHEREY-NAGEL). The filter was saturated in order to avoid adsorption during the filtration. The first 25 mL of filtrate was discarded. The filtration was interrupted for ca. 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 was discarded. The following filtrate, i.e. the saturated solution, was used as highest loading level in the test and for preparation of further loading rates by diluting with dilution water. During filtration, the filter was always kept covered.
Test loadings
5 loading rates of the test item in a geometric series with a separation factor of 2.5, prepared by diluting the saturated solution with dilution water, were tested as follows:
1.02 - 2.56 - 6.40 - 16.0 - 40.0 - 100 % of the saturated solution.
The loading levels are based on the results of a preliminary range finding test (non GLP)
Test concentrations (geometric mean measured)
19.7 – 51.0 – 129 – 330 – 869 – 2299 µg/L
Control
Six replicates (without test item) were tested under the same test conditions as the test replicates. - Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- TEST ORGANISM
- Common name: Desmodesmus subspicatus Chodat
- Strain: SAG 86.81
- 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 were prepared every month on Z-Agar. Light intensity amounted to 35 - 70 µE x m-2 x s-1 for 24 h per day. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- No
- Hardness:
- not specified
- Test temperature:
- min.: 21.0 max.: 22.0, mean value: 21.5 °C
- pH:
- Geometric mean measured test item concentration pH-value
[µg/L] Start; 0 hours End; 72 hours
2299 7.39 7.38
869 7.47 7.40
330 7.54 7.55
129 7.64 7.66
51.0 7.73 7.78
19.7 7.85 7.92
Control 7.90 8.13 - Dissolved oxygen:
- Not measured
- Salinity:
- Not measured, freshwater conditions
- Nominal and measured concentrations:
- See window "Any other information on materials and methods incl. tables"
- 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 2 - 5 x 10^3 cells/mL, Actual: mean 4847 cells/mL
- Control end cells density: Mean 1358509 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 x 6 H2O 12
CaCl2 x 2 H2O 18
MgSO4 x 7 H2O 15
KH2PO4 1.6
FeCl3 x 6 H2O 0.064
Na2EDTA x 2 H2O 0.1
H3BO3 0.185
MnCl2 x 4 H2O 0.415
ZnCl2 3 x 10^-3
Na2MoO4 x 2 H2O 7 x 10^-3
CoCl2 x 6 H2O 1.5 x 10^-3
CuCl2 x 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: 70.6 µE x m^-2 x s^-1,
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. - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 129 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 330 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 263 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 186 - 311
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 300 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 220 - 322
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 374 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 344 - 561
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 129 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 330 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 134 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Remarks on result:
- other: 116 - 160
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 163 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Remarks on result:
- other: 143 - 187
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 229 µg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- other: yield
- Remarks on result:
- other: 207 - 250
- 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):
- The toxicity of potassium dichromate (SIGMA, batch number MKBQ9179V, purity 100.0 %, CAS RN 7778-50-9) to the unicellular freshwater green alga Desmodesmus subspicatus was determined over a period of 72 hours from 2015-10-19 to 2015-10-22. The reference item toxicity is in the valid range following test facility SOPS.
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 0.685 0.671 ± 0.401
95 % confidence interval 0.603 - 0.794
Yield Inhibition
EyC50 0.362 0.320 ± 0.139
95 % confidence interval 0.328 - 0.398
SD = Standard deviation - Reported statistics and error estimates:
- EC-values and statistical analyses
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 of the EC10-, EC20- and EC50-values was carried out using standard procedures.
NOEC- and LOEC-values
The NOEC / LOEC was determined by calculation of statistical significance of growth rate and yield. The following statistical tests were conducted:
Growth rate and yield: Shapiro-Wilk’s test on normal distribution was done with a significance level 0.01.
Growth rate and yield: Levene’s test on variance homogeneity was done with a significance level 0.01.
Growth rate and yield: Monotonicity of concentration/response was done by trend analysis by contrasts (significance level 0.05).
Growth rate: Step-down Jonckheere-Terpstra’s test on significant differences was done with a significance level 0.05.
Yield: Multiple Sequentially-rejective Welsh-t-test after Bonferroni-Holm was done with a significance level 0.05.
Software
The data for the tables in this report were computer-generated and have been rounded for presentation from the fully derived data. Consequently, if calculated manually based on the given data minor deviations may occur from these figures.
Calculations were carried out using software
- Excel, MICROSOFT CORPORATION
- SigmaPlot, SPSS INC.
- ToxRat, ToxRat Solutions GmbH - Validity criteria fulfilled:
- yes
- Conclusions:
- In this study, the substance was found to inhibit the growth of the freshwater green alga Desmodesmus subspicatus after 72 hours with the following effect values (based on geometric mean measured test item concentrations): The EC50-values with 95 % confidence intervals for both inhibition of growth rate (ErC50) and yield (EyC50) after 72 hours were 374 (344 – 561) and 229 (207 – 250) µg/L. The NOEC-values for inhibition of growth rate and yield after 72 hours were for both 129 µg/L.
- Executive summary:
Summary
The toxicity of the substance to the unicellular freshwater green alga Desmodesmus subspicatus was determined according to the principles of OECD 201 (2011) and Council Regulation (EC) No. 761/2009/Method C.3 (2009) at the test facility from 2016-02-01 to 2016-02-19 with the definitive exposure phase from 2016-02-16 to 2016-02-19.The aim of the study was to assess the effects on growth rate and yield over a period of 72 hours.
The study was conducted under static conditions with an initial cell density of 4847 cells/mL. Based on a preliminary range finding test a saturated solution with a nominal loading of 100 mg test item/L was prepared in dilution water. The dispersion was stirred for 24 hours with approximately 1100 rpm at room temperature. At the end of the stirring period, undissolved test item was removed by membrane filtration (membrane filter 0.45 µm, RC, Macherey-Nagel). The saturated solution and further five dilution levels out of the saturated solution were tested in a geometrical series with a dilution factor of 2.5: 1.02 - 2.56 - 6.40 - 16.0 - 40.0 - 100 % of the saturated solution corresponding to geometric mean measured concentration levels of: 19.7 – 51.0 – 129 – 330 – 869 – 2299 µg/L.Three replicates were tested for each test item concentration and six replicates for the control. Environmental conditions were determined to be within the acceptable limits.
All test concentrations of the test item and the control were analytically verified by LC-MS/MS analysis at the start of the exposure (0 hours) and the end of the exposure (72 hours).
The measured concentrations of the test item at the start of the exposure were in the range of 21.8 to 2564 µg/L. At the end of the exposure, they were in the range of 17.8 to 2062 µg/L.
All effect values given are based on the geometric mean measured test item concentrations.NOEC, LOEC, EC - values of the test item (0-72 hours)
based on the geometric mean measured test item concentration [µg/L]
Growth Rate Inhibition
geometric mean measured test item concentration [µg/L]
NOEC
129
LOEC
330
ErC10
263 (186 – 311)
ErC20
300 (220 – 322)
ErC50
374 (344 – 561)
Inhibition of Yield
geometric mean measured test item concentration [µg/L]
NOEC
129
LOEC
330
EyC10
134 (116 – 160)
EyC20
163 (143 – 187)
EyC50
229 (207 – 250)
Reference
Cell Densities
Geometric mean measured test item concentration |
Replicate |
Cell density [cells/mL] |
|||
[µg/L] |
No. |
0 hours |
24 hours |
48 hours |
72 hours |
2299 |
1 |
4847 |
10658 |
684* |
684* |
2 |
4847 |
10802 |
1763 |
684* |
|
3 |
4847 |
13260 |
1768 |
684* |
|
Mean |
4847 |
11573 |
1405 |
684* |
|
869 |
1 |
4847 |
20110 |
6107 |
684* |
2 |
4847 |
22323 |
4111 |
4313 |
|
3 |
4847 |
21073 |
7016 |
2081 |
|
Mean |
4847 |
21169 |
5745 |
2359 |
|
330 |
1 |
4847 |
25017 |
37149 |
137665 |
2 |
4847 |
30063 |
46756 |
237535 |
|
3 |
4847 |
29005 |
75032 |
386295 |
|
Mean |
4847 |
28028 |
52979 |
253832 |
|
129 |
1 |
4847 |
39747 |
235510 |
1172711 |
2 |
4847 |
35442 |
254396 |
1288837 |
|
3 |
4847 |
36841 |
237704 |
1257159 |
|
Mean |
4847 |
37343 |
242537 |
1239569 |
|
51.0 |
1 |
4847 |
43076 |
305522 |
1352239 |
2 |
4847 |
39155 |
338955 |
1449315 |
|
3 |
4847 |
43744 |
310814 |
1379899 |
|
Mean |
4847 |
41992 |
318430 |
1393818 |
|
19.7 |
1 |
4847 |
40921 |
323081 |
1374175 |
2 |
4847 |
42417 |
291812 |
1320490 |
|
3 |
4847 |
39699 |
312497 |
1294321 |
|
Mean |
4847 |
41012 |
309130 |
1329662 |
|
Control |
1 |
4847 |
34576 |
297248 |
1276377 |
2 |
4847 |
34672 |
288955 |
1503625 |
|
3 |
4847 |
36211 |
292899 |
1653617 |
|
4 |
4847 |
38482 |
317279 |
1264592 |
|
5 |
4847 |
44793 |
254310 |
1238326 |
|
6 |
4847 |
30934 |
291225 |
1214514 |
|
Mean |
4847 |
36611 |
290319 |
1358509 |
*)
values below the limit of quantification (1368 cells/mL) are given as ½
limit of quantification
Evaluation after 72 hours
Statistically significant differences of growth rates and yield compared to
control values are marked (+), not significant differences are marked (-).
Geometric mean measured test item concentration |
Replicate |
Growth rate |
Inhibition of growth rate |
Yield |
Inhibition of yield |
||
[µg/L] |
No. |
[d-1] |
[%] |
[cells/mL] |
[%] |
||
2299 |
1 |
|
-0.653* |
100 |
|
-4163* |
100 |
2 |
|
-0.653* |
100 |
|
-4163* |
100 |
|
3 |
|
-0.653* |
100 |
|
-4163* |
100 |
|
Mean |
(+) |
-0.653* |
100 |
(+) |
-4163* |
100 |
|
869 |
1 |
|
-0.653* |
100 |
|
-4163* |
100 |
2 |
|
-0,039* |
100 |
|
-534* |
100 |
|
3 |
|
-0,282* |
100 |
|
-2766* |
100 |
|
Mean |
(+) |
-0.324* |
100 |
(+) |
-2487* |
100 |
|
330 |
1 |
|
1.12 |
41 |
|
132818 |
90 |
2 |
|
1.30 |
31 |
|
232688 |
83 |
|
3 |
|
1.46 |
22 |
|
381448 |
72 |
|
Mean |
(+) |
1.29 |
31 |
(+) |
248985 |
82 |
|
129 |
1 |
|
1.83 |
2 |
|
1167864 |
14 |
2 |
|
1.86 |
1 |
|
1283990 |
5 |
|
3 |
|
1.85 |
1 |
|
1252312 |
7 |
|
Mean |
(-) |
1.85 |
2 |
(-) |
1234722 |
9 |
|
51.0 |
1 |
|
1.88 |
0 |
|
1347392 |
0 |
2 |
|
1.90 |
-1 |
|
1444468 |
-7 |
|
3 |
|
1.88 |
0 |
|
1375052 |
-2 |
|
Mean |
(-) |
1.89 |
-1 |
(-) |
1388971 |
-3 |
|
19.7 |
1 |
|
1.88 |
0 |
|
1369328 |
-1 |
2 |
|
1.87 |
0 |
|
1315643 |
3 |
|
3 |
|
1.86 |
1 |
|
1289474 |
5 |
|
Mean |
(-) |
1.87 |
0 |
(-) |
1324815 |
2 |
|
Control |
1 |
|
1.86 |
|
|
1271530 |
|
2 |
|
1.91 |
|
|
1498778 |
|
|
3 |
|
1.94 |
|
|
1648770 |
|
|
4 |
|
1.86 |
|
|
1259745 |
|
|
5 |
|
1.85 |
|
|
1233479 |
|
|
6 |
|
1.84 |
|
|
1209667 |
|
|
Mean |
|
1.88 |
|
|
1353662 |
|
Negative values = growth stimulation
*) calculated values, considered to be 0.
Description of key information
OECD 201 (Desmodesmus subspicatus; static; mean measured; RL 1):
NOEC (72 h; growth rate): 129 µg/L;
LOEC (72 h; growth rate): 330 µg/L;
EC10 (72 h; growth rate): 263 µg/L (95% CI: 186 - 311);
EC50 (72 h; growth rate): 374 µg/L (95% CI: 344 - 561).
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 374 µg/L
- EC10 or NOEC for freshwater algae:
- 263 µg/L
Additional information
The toxicity of the substance to the unicellular freshwater green alga Desmodesmus subspicatus was determined according to the principles of OECD 201 (2011) and Council Regulation (EC) No. 761/2009/Method C.3 (2009) compliant with GLP (RL1; Noack, 2016).
The study was conducted under static conditions with an initial cell density of 4847 cells/mL. Based on a preliminary range finding test a saturated solution with a nominal loading of 100 mg test item/L was prepared in dilution water. After stirring for 24 hours at room temperature undissolved test item was removed by membrane filtration. The following 6 concentration levels of the test item were applied, given in % of the saturated solution (diluted with test water): 1.02 - 2.56 - 6.40 - 16.0 - 40.0 - 100 %. These concentration levels corresponded to the following geometric mean measured concentrations: 19.7 – 51.0 – 129 – 330 – 869 – 2299 µg/L. Three replicates were tested for each test item concentration and six replicates for the control. Environmental conditions were determined to be within the acceptable limits.
All test concentrations of the test item and the control were analytically verified by LC-MS/MS analysis at the start of the exposure (0 hours) and the end of the exposure (72 hours). The measured concentrations of the test item at the start of the exposure were in the range of 21.8 to 2564 µg/L. At the end of the exposure, they were in the range of 17.8 to 2062 µg/L. pH values within treatment groups were between 7.38 and 7.92, pH for the controls was between 7.9 and 8.13.
All effect values given are based on the geometric mean measured test item concentrations:
NOEC (72 h; growth rate): 129 µg/L;
LOEC (72 h; growth rate): 330 µg/L;
EC10 (72 h; growth rate): 263 µg/L (95% CI: 186 - 311);
EC50 (72 h; growth rate): 374 µg/L (95% CI: 344 - 561).
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