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EC number: 947-299-2 | 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:
- 2017-10-03 to 2017-11-17
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Remarks:
- GLP study performed according to the OECD 201 guideline. All validity criteria were successful.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Deviations:
- no
- Principles of method if other than guideline:
- Not applicable.
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Signed on 2017-11-22
- Specific details on test material used for the study:
- None
- Analytical monitoring:
- yes
- Details on sampling:
- In order to verify maintenance of the WAFs during the ecotoxicological testing, chemical analyses of test item were performed in the test solutions (biotic and abiotic replicates):
- at the beginning of the test (T = 0h)
- at the end of the test (T = 72h)
Schedule of analysis at initial time T0h:
- WAF Control Abiotic
- WAF 100.0 mg/L Abiotic
Schedule analysis at final time T72h:
- WAF Control Biotic
- WAF 100.0 mg/L Biotic
- WAF Control Abiotic
- WAF 100.0 mg/L Abiotic - Vehicle:
- no
- Details on test solutions:
- TEST ITEM PREPARATION
the test item was not soluble in the test medium, a special solubilisation protocol was carried out before the experimentation, according to the OECD n°23 guidance document on aquatic toxicity testing of difficult substances and mixtures.
Once having tried different methods for the preparation of the WAFs in the preliminary study, the following protocol was selected for the final test: this method consisted in stirring the test item nominal concentration 100 mg/L in test medium during 24 hours, in the dark at 20 ± 2°C. The test solution (100 mg/L) was then filtered (cellulose acetate with a mesh size of 0.45 µm) to remove suspended particles: the solution obtained was named WAF (Water Accommodated Fraction).
TEST SOLUTIONS
Two non-GLP preliminary studies (with and without solvent) have been performed in order to select the method for the test solution preparation and the concentrations to be used in the final test. Based on these results, the final test was performed at the limit test item concentration of 100 mg/L, and without solvent.
For the non-GLP preliminary test with solvent, DMSO has been determined as the most suitable solvent to be used. For stock solution at 100 g test item /L DMSO, 4 loading rates 1; 10; 50 and 100 mg/L were prepared before test with algae. During this study, it has not been possible to solubilize the test item at the maximal solvent concentration recommended in the OECD 23 guideline (100 mg/L or 0.1 mL/L). The solvent concentrations used were above this limit. Therefore, this method could not be selected for the final test.
The final test contained:
- Control: 6 biotic replicates + 2 abiotic replicates
- Concentration 100 mg/L: 6 biotic replicates + 2 abiotic replicates - Test organisms (species):
- Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata
- Strain: chlorococcal order strain number CCAP 278.4
The algae cultures are stored in the laboratory on a solid medium (LC/agar Petri plates) and in a liquid medium with regular sub-culturing (every week). - Test type:
- static
- Water media type:
- freshwater
- Remarks:
- The medium was prepared from concentrated stock solutions in ultrapure water as described in the guideline OECD 201.
- Limit test:
- yes
- Total exposure duration:
- 72 h
- Remarks on exposure duration:
- None.
- Post exposure observation period:
- None.
- Hardness:
- No data.
- Test temperature:
- 22.8 °C – 23.3°C
- pH:
- 8.0 - 8.1
- Dissolved oxygen:
- No data.
- Salinity:
- No data.
- Conductivity:
- No data.
- Nominal and measured concentrations:
- Limit test: 100 mg test item /L (nominal, loading rate)
- Details on test conditions:
- TEST SYSTEM
In agreement with the study monitor and according to the results from the preliminary tests, this study was conducted with one concentration level: 100.0 mg/L: Limit test, WAF conditions.
A growing exponential phase inoculum of the freshwater unicellular alga Pseudokirchneriella subcapitata was added in order to obtain an initial concentration of 10^4 cells/mL at the start of the test: 100 mL of algal cultures grown in 250 mL erlenmeyer flasks which were placed in a cooling incubator at a temperature in the range 21 to 24°C controlled at ± 2°C and under a constant shaking fixed at 120 rpm and a continuous illumination comprised between 4 440 and 8 880 lux.
The algal growth inhibition in relation to the control was evaluated over a period of 72 hours. The algal cells per unit volume were measured during the test at 24, 48 and 72 hours with a particle counter (Beckman Coulter Z2) and/ or under microscope with Malassez counting cell.
The pH of each test concentration was measured at the start and at the end of the test. The pH of the control medium had not increased by more than 1.5 units during the test.
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS.
The medium was prepared from concentrated stock solutions in ultrapure water as described in the guideline
Algal dilution water
Theoretical composition
NH4Cl 15 mg/L
MgCl2, 6H20 12 mg/L
CaCl2, 2H2O 18 mg/L
MgSO4,7H20 15 mg/L
KH2PO4 1.6 mg/L
FeCl3, 6H2O 0.08 mg/L
Na2EDTA, 2H2O 0.1 mg/L
H3BO3 0.185 mg/L
MnCl2, 4H2O 0.415 mg/L
ZnCl2 3 µg/L
CoCl2, 6 H20 1.5 µg/L
CuCl2, 2H2O 10-2 µg/L
Na2MoO4, 2H2O 7 µg/L
NaHCO3 50 mg/L
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: No
- Light intensity and quality: 6461 Lux
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Beckman Coulter Z2 and Malassez counting cell
TEST CONCENTRATIONS
- Test concentrations: control, 1, 10 and 100 mg/L with the same method of WAF preparation than the definitive test.
- Another non-GLP preliminary test with solvent conditions was also performed. Acetone has been determined as the most suitable solvent to be used. For stock solution at 100 g test item /L acetone, 4 loading rates 1; 10; 50 and 100 mg/L were prepared before acute immobilisation test with daphnids. During this study, it has not been possible to solubilize the test item at the maximal solvent concentration recommended in the OECD 23 guideline (100 mg/L or 0.1 mL/L). The solvent concentrations used were above this limit. Therefore, this method cannot be selected for the final test. - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- > 100 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- other: Yield inhibition
- Results with reference substance (positive control):
- The last algal growth inhibition test performed with the reference item K2Cr2O7 (September 18 2017) gave an ErC50(0-72h) = 1.11 mg/L (0.98 – 1.24), indicating that the algae sensitivity is correct (within the accepted interval of 0.65 to 1.73 mg/L fixed by the International Standard NF EN ISO 8692 from Mai 2012 as mentioned in the OECD Guideline) and in accordance with previous data obtained by the laboratory.
- Reported statistics and error estimates:
- Not applicable.
- Validity criteria fulfilled:
- yes
- Conclusions:
- The results obtained for the test item “TREE MOSS CONCRETE” on the toxicological endpoint “inhibition of growth rate” indicate an ErL50-(0-72h) higher than the highest tested loading rate, i.e. 100 mg/L, based on nominal test item loading rate.
- Executive summary:
The aim of this study was to determine the effects of the test item "TREE MOSS CONCRETE" on the growth of the unicellular green alga Pseudokirchneriella subcapitata, according to the OECD 201 guideline (March 2006, corrected July 2011). The algae were exposed to Water Accommodated Fractions (WAFs) of the test item, following the“Guidance document on aquatic toxicity testing of difficult substances and mixtures” (OECD No. 23).
Following the results of a preliminary study, a limit test at a nominal loading rate of 100 mg test item/L was performed, in order to demonstrate that the test item has no significant effect on the test organism at this test loading rate or that EL50 at 72h is higher than 100 mg/L (loading). The effects on the growth of the algae was determined by cell counting daily during 72 hours.
In order to check the stability of the WAFs in the test solutions, concentration of dissolved organic material in the control and the WAFs was checked by TOC analysis at start (t=0h) and at the end of the test (t=72h). These non-specific chemical analyses were performedby Carbon Analyzer.
The table below summarizes the achieved results and the variation between T0 and T72 hours.
Nominal test item loading rate
mg “NPOC”/L in algae solutions
Variation between T0 and T72 hrs (%)
Results at T0 hour
Results at T72 hours
WAF Control Biotic
/
5.02
NA
WAF 100 mg/L Biotic
/
4.34
NA
WAF Control Abiotic
< 2 (0.56)*
3.02
NA
WAF 100 mg/L Abiotic
< 2 (0.85)*
3.26
NA
*for information
NA: not applicable
All measured values were lower than 2 mg/L (=LOQ) and are notified for information (in brackets). The slight increase after 72 hours of exposition was mainly due to the gaseous exchange.
TOC analyses indicated that organic compounds were found below the LOQ in the control and in the WAFs at 100 mg/L. Therefore, the maintenance of the WAFs throughout the test, cannot be accurately evaluated. Nevertheless, since WAFs are by definition a complex mixture and since the test item is an UVCB substance, the results are based on the nominal test loading rates.
The results of this study are summarized below:
- Test item: TREE MOSS CONCRETE
- Test system: Pseudokirchneriella subcapitata
- Exposure duration: 72 hours
- Test concentrations: Control - 100 mg/L
- Incubator: Temperature 22.8 – 23.3 °C ; shaking at 120 rpm; light intensity: 6461 Lux
- Measurement of cell growth: Beckman Coulter Z2 and Malassez counting cell
- Validity criteria: In the control. the increase of the biomass during 72 hours was determined to be higher than factor of 16.The coefficients of variation of daily section-by-section specific growth rateand ofaverage specific growth ratewere determined respectively to be less than 35% and 7% in the control. - The validity criteria were successfully fulfilled.
- Results: The results obtained for thetest item “TREE MOSS CONCRETE”on the toxicological endpoint “inhibition of growth rate” indicate an ErL50-(0-72h) higher than the highest tested loading rate, i.e. 100 mg/L, based on nominal test item loading rate.
The results obtained for the test item “TREE MOSS CONCRETE”on the toxicological endpoint “Yield inhibition” indicate anEyL50-(0-72h)higher than the highest tested loading rate, i.e. 100 mg/L, based on nominal test item loading rate.
Reference
Validity criteria for the test
|
Criteria for OECD 201 |
Validity criteria conformity |
Increase biomass during 72 hours |
Factor of at least 16 |
Yes (164.7) |
Daily section-by-section specific growth rate CV |
< 35% |
Yes (17.2%) |
Average specific growth rate CV during 72 hours |
< 7% |
Yes (1.0%) |
All the validity criteria were successfully validated.
Algal growth inhibition
Raw data,i.e. algal concentrations values expressed as cell number *104per mL of test solutions after 24, 48 and 72 hours of incubation are indicated in the following table.
Nominal test item loading rate |
[algae] *104/mL at 24 hours |
[algae] *104/mL at 48 hours |
[algae] *104/mL at 72 hours |
|||||||
Count. |
Average |
CV |
Count. |
Average |
CV |
Count. |
Average |
CV |
||
WAF Control |
a |
5,1 |
4.5 |
11.7 |
22,3 |
22.1 |
8.1 |
153,8 |
164.7 |
5.0 |
b |
4,5 |
21,4 |
162,4 |
|||||||
c |
4,3 |
21,1 |
169,1 |
|||||||
d |
4,0 |
24,3 |
157,2 |
|||||||
e |
5,2 |
23,8 |
172,8 |
|||||||
f |
4,0 |
19,5 |
173,1 |
|||||||
WAF 100 mg/L |
a |
4,4 |
4.0 |
16.5 |
26,5 |
22.8 |
20.7 |
148,7 |
149.3 |
20.7 |
b |
3,5 |
19,1 |
125,1 |
|||||||
c |
4,1 |
30,4 |
134,2 |
|||||||
d |
3,4 |
18,1 |
117,5 |
|||||||
e |
3,4 |
21,5 |
171,3 |
|||||||
f |
5,0 |
21,2 |
199,1 |
CV: Coefficient of variation (%)
The coefficients of variation for section-by-section specific growth rates in the control cultures are presented in the following table to calculate one of the validity criterion: average value of coefficient of variation (Average CV).
Control replicates |
Specific growth rate |
|||||||||||||
0-24h |
Average |
CV (%) |
24 -48h |
Average |
CV (%) |
48 -72h |
Average |
CV (%) |
0 - 72h |
Average |
CV per replicate (%) |
Average CV |
||
WAF Control |
a |
1,629 |
1,502 |
7,7 |
1,475 |
1,589 |
7,2 |
1,931 |
2,012 |
5,6 |
1,679 |
1,701 |
13,8 |
17.2% |
b |
1,504 |
1,559 |
2,027 |
1,697 |
16,9 |
|||||||||
c |
1,459 |
1,591 |
2,081 |
1,710 |
19,2 |
|||||||||
d |
1,386 |
1,804 |
1,867 |
1,686 |
15,5 |
|||||||||
e |
1,649 |
1,521 |
1,982 |
1,717 |
13,9 |
|||||||||
f |
1,386 |
1,584 |
2,183 |
1,718 |
24,2 |
CV: Coefficient of variation (%)
Chemical analysis of algal solutions
The table below summarizes the results achieved and the variation between T0 and T72 hours.
Nominal test item loading rate |
mg “NPOC”/L in algae solutions |
Variation between T0 and T72 hrs (%) |
|
Results at T0 hour |
Results at T72 hours |
||
WAF Control Biotic |
/ |
5.02 |
NA |
WAF 100 mg/L Biotic |
/ |
4.34 |
NA |
WAF Control Abiotic |
< 2 (0.56)* |
3.02 |
NA |
WAF 100 mg/L Abiotic |
< 2 (0.85)* |
3.26 |
NA |
*(for information)
pH values and observations
Nominal test item loading rate (mg/L) |
pHat the start (biotic replicates) |
pHafter 72 hours (biotic replicates) |
WAF Control |
8.0 |
7.4 |
WAF 100 |
7.9 |
7.4 |
Description of key information
The results obtained for the test item on the toxicological endpoint “inhibition of growth rate” indicate an ErL50-(0-72h) higher than the highest tested loading rate, i.e. 100 mg/L, based on nominal test item loading rate.
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 100 mg/L
Additional information
The results of this study are summarized below:
- Test item: TREE MOSS CONCRETE
- Test system: Pseudokirchneriella subcapitata
- Exposure duration: 72 hours
- Test concentrations: Control - 100 mg/L (limit test)
- Incubator: Temperature 22.8 – 23.3 °C ; shaking at 120 rpm; light intensity: 6461 Lux
- Measurement of cell growth: Beckman Coulter Z2 and Malassez counting cell
Results: The results obtained for the registered substance on the toxicological endpoint “inhibition of growth rate” indicate an ErL50-(0-72h)higher than the highest tested loading rate,i.e. 100 mg/L, based on nominal test item loading rate.
The results obtained for thetest item “TREE MOSS CONCRETE” on the toxicological endpoint “Yield inhibition” indicate an EyL50-(0-72h) higher than the highest tested loading rate, i.e. 100 mg/L, based on nominal test item loading rate.
Validity criteria: In the control, the increase of the biomass during 72 hours was determined to be higher than factor of 16.The coefficients of variation ofdaily section-by-section specific growth rateand ofaverage specific growth ratewere determined respectively to be less than 35% and 7% in the control. The validity criteria were successfully fulfilled.
This study respected the requirements of the test guideline. The study was considered acceptable for that endpoint.
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