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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:
- From 1996-03-01 to 1996-07-26
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Although a guideline study (OECD 201 & EU Method C.3) conducted according to the principles of Good Laboratory Practice, the purity, batch number and expiry date of the test substance was not supplied and its stability under the test conditions was not determined. Also, verification of exposure concentrations during the study was not undertaken and the test results are based on the nominal concentrations used to prepare the test media.
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Version / remarks:
- 1984
- Deviations:
- yes
- Remarks:
- temperature of test area slightly exceeded maximum
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Version / remarks:
- 1992
- Deviations:
- no
- GLP compliance:
- yes
- Specific details on test material used for the study:
- - Substance type: technical material
- Physical state: a fine cream-coloured cohesive powder
- Storage condition of test material: at ambient temperature
- Other: Test concentrations quoted in the report refer to the test material as received; no allowance has been made for a purity of less than 100% - Analytical monitoring:
- no
- Vehicle:
- yes
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test medium was prepared from an aqueous dispersion (nominally 111 mg/Litre) in which the test material, mixed with acetone, had been added to OECD medium; to aid dispersion it was subjected to twenty minutes of ultrasound treatment. This aqueous dilution was then shaken overnight in the dark before being centrifuged at 3000 rpm for 15 minutes to remove the bulk of the particulate material.
- Controls: algal suspension alone and a solvent control containing algal suspension and acetone.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.1 mL/L
Additional comment on medium preparation (information not presented in test report):
The test substance was multi-component and poorly soluble in water so the test media were prepared using techniques based on those employed for generating water accommodated fractions (WAFs). An auxiliary solvent was also used to aid dissolution and dispersion of the test substance. The physical methods employed were ultrasound and prolonged vigorous stirring. The formulation methods employed were designed to maximise the concentrations of the test substance in solution and therefore bioavailable to the test organisms. To avoid exposing the test organisms to large amounts of insoluble test material, the media used in the fish and Daphnia tests were left to equilibrate overnight before use and then the aqueous fraction was removed mid-vessel and used to fill the test vessels; for the algal study, the insoluble fraction of the test substance was removed by centrifugation. Although a concentrated stock was prepared for the fish test (to facilitate ultrasound treatment), the entire contents of the stock prepared for each batch of test medium was diluted to the required final volume, so the composition of the test mixture would not have been altered. Although the use of an auxiliary solvent may have an effect on the proportions of the different components dissolved, the amount used (0.1 mL/L) was very small and its impact is thought to have been negligible. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Source (laboratory, culture collection): Culture Collection of Algae and Protozoa (CCAP), Freshwater Biological Association, Ferry House, Ambleside, Cumbria, UK.
- Age of inoculum (at test initiation): 2 months
- Method of cultivation: Liquid cultures for the test inoculum were established by inoculation of OECD medium (50 or 100 mL) with cells aseptically removed from slope cultures. Subsequently, subcultures were prepared from these liquid cultures by the aseptic transfer of aliquots into fresh OECD medium, which was then incubated for several days, until the required cell density had been achieved.
ACCLIMATION
- Culturing media and conditions (same as test or not): Same as test - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 72 h
- Test temperature:
- 21.6 to 25.3 °C
- pH:
- 7.2 to 8.4
- Nominal and measured concentrations:
- Nominal: 100 mg/L
- Details on test conditions:
- TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): loosely plugged with non-absorbent cotton wool
- Material, size, headspace, fill volume: 250 mL conical flasks, filled with 90 mL test medium
- Initial cells density: 1E+04 cells/mL
- Control end cells density: OECD control: 65.8-115 E+04 cells/mL; solvent control: 58.3-123 cells/mL
- No. of organisms per vessel: 1E+04 cellls per mL
- No. of vessels per concentration (replicates): 7(6 were incubated, the other used for temperature and pH at start)
- No. of vessels per control (replicates): 7 (6 were incubated, the other used for temperature and pH at start)
- No. of vessels per vehicle control (replicates): 7 (6 were incubated, the other used for temperature and pH at start)
GROWTH MEDIUM
- Standard medium used: yes (OECD)
OTHER TEST CONDITIONS
- Sterile test conditions: yes (aseptic conditions)
- Adjustment of pH: no
- Photoperiod: Continuous illumination was supplied by four, horizontallymounted, three-foot 30 watt "cool- white" fluorescent tubes.
- Light intensity and quality: 7700 lux (measurements taken with a Radiospares luxmeter during the definitive test showed 1900 - 1950 lux when a
single tube was illuminated).
- Salinity (for marine algae): N/A
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Haemocytometer (Improved Neubauer).
TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0.1, 1, 10 and 100 mg/Litre
- Results used to determine the conditions for the definitive study: Several tests were conducted at nominal concentrations in the range 0.1 to 100 mg/L in which algal growth was adversely affected but the effects were neither concentration-related or reproducible. Following a test in which algae were exposed to centrifuged preparations containing test item it was concluded that the variability observed was due to particulate material in the test media. Therefore, 100mg/Litre chosen for definitive test. - Reference substance (positive control):
- no
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: The median effect concentration (ErC50 or EbC50) could not be identified but it must be at least equal to the limit of aqueous solubility of the test material.
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: The no-observed-effect concentration (NOEC) could not be identified but it must be at least equal to the limit of aqueous solubility of the test material.
- Details on results:
- - Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): None
- Any stimulation of growth found in any treatment: No - Reported statistics and error estimates:
- A statistical comparison of the average specific growth rates in control and test cultures was carried out using Dunnett's multicomparison test.
In this test, a multiple t-test compares each treatment with the control using a common estimate of experimental error, and then uses Dunnett's
t-Statistic tables to assess significance at the 95% level of probability. -
Calculation of growth rates Concentration-effect
relationships were calculated by comparing growth rates in control and
test cultures in the following way. The average specific growth rate
(u) for individual cultures was calculated from the following
relationship:
u = 1n (Nn) - 1n (N0)
tn- t0 Where 1n (Nn) = natural logarithm of cell number at time tn; 1n (N0) = natural logarithm of cell number at time t0; t0= nominal measurement at the beginning of the test; tn= time of nth measurements after the beginning of the test. The average specific growth rate of the test culture was expressed as a percentage of the solvent control, and subtracted from 100 to give the percentage effect. The ErC50 could not be calculated because no inhibition of algal growth occurred. -
Calculation of the areas under the growth curves Biomass
was calculated by determining the areas under the growth curves of
control and test cultures in the following way:
- Validity criteria fulfilled:
- yes
- Conclusions:
- Exposure of Selenastrum capricornutum to an aqueous mixture of test item, derived from a preparation at an initial nominal level of100 mg/Litre, did not result in a significant reduction of either the average specific growth rates or the mean biomass values compared to control cultures (p >0.05). Neither the median effect concentration (ErC50 or EbC50) nor the no-observed-effect concentrations could be identified but both must be at least equal to the limit of aqueous solubility of the test material.
- Executive summary:
A study was conducted to investigate the effects of test item on the growth of the unicellular green alga Selenastrum capricornutum over a 72 hour period. The study was conducted in mineral salts medium at temperatures in the range 21.6 to 25.3 °C in an illuminated orbital incubator using methods based on OECD Procedure 201 and the Annex to EC Directive 92/69/EEC, Part C3. The exposure concentration (100mg/Litre) was not verified during the limit test because attempts to develop an appropriate method of chemical analysis were unsuccessful. The formulation methods employed are considered to have given rise to the maximum attainable exposure concentration of test item under the conditions of the test. Exposure of Selenastrum capricornutum to an aqueous mixture of the test substance did not result in a significant reduction of either the average specific growth rates or the mean biomass values compared to solvent controls (p >0.05). Neither the median effect concentration (ErC50 or EbC50) nor the no-observed-effect concentrations could be identified but both must be at least equal to the limit of aqueous solubility of the test material.
Reference
Limit test - algal growth
The results of cell counts in control and test flasks are given in Table 1 below.
The average specific growth rates and biomass in control and test cultures are given in Table 2 below.
Exposure of Selenastrum capricornutum to an aqueous mixture of test item, derived from a preparation at an initial nominal concentration of 100 mg/Litre, did not result in a significant reduction of either the average specific growth rates or the mean biomass values compared to solvent control cultures (p > 0.05). Neither the median effect concentration (ErC50 or EbC50) nor the no-observed-effect concentrations could be identified but both must be at least equal to the limit of aqueous solubility of the test material.
Table 1.
Cell numbers in individual control and test cultures
Nominal test material conc. (mg/L) |
Flask number |
Cell numbers per mL (x104) |
||
24 hours |
48 hours |
72 hours |
||
0 (OECD control) |
1 |
6.38 |
33.3 |
86.3 |
2 |
4.88 |
36.0 |
71.4 |
|
3 |
5.13 |
29.5 |
74.1 |
|
4 |
7.25 |
38.6 |
115 |
|
5 |
5.50 |
32.3 |
65.8 |
|
6 |
4.75 |
29.0 |
76.4 |
|
|
||||
0 (solvent control) |
8 |
6.00 |
31.4 |
71.3 |
9 |
6.75 |
25.6 |
58.5 |
|
10 |
3.63 |
27.8 |
60.3 |
|
11 |
5.00 |
36.8 |
123 |
|
12 |
5.00 |
23.9 |
58.6 |
|
13 |
5.63 |
31.4 |
87.3 |
|
|
||||
100 |
15 |
5.25 |
41.1 |
106 |
16 |
4.38 |
25.9 |
59.1 |
|
17 |
5.25 |
24.6 |
54.8 |
|
18 |
5.00 |
33.1 |
97.5 |
|
19 |
5.00 |
21.5 |
49.4 |
|
20 |
4.50 |
22.8 |
51.0 |
Table 2.
Average specific growth rates and biomass (0-72 hours) in control and test cultures
Nominal test material conc. (mg/L) |
Flask number |
Average specific growth rate x10-2 |
Mean x10-2 |
Biomass x10-4 |
Mean x10-4 |
0 (OECD control) |
1 |
6.191 |
6.088 |
1928 |
1848 |
2 |
5.928 |
1778 |
|||
3 |
5.980 |
1660 |
|||
4 |
6.590 |
2420 |
|||
5 |
5.815 |
1637 |
|||
6 |
6.022 |
1667 |
|||
|
|||||
0 (solvent control) |
8 |
5.926 |
5.969 |
1693 |
1694 |
9 |
5.651 |
1418 |
|||
10 |
5.694 |
1418 |
|||
11 |
6.684 |
2419 |
|||
12 |
5.654 |
1337 |
|||
13 |
6.207 |
1876 |
|||
|
|||||
100 |
15 |
6.477 |
5.824 (2%) |
2324 |
1569 (7%) |
16 |
5.666 |
1376 |
|||
17 |
5.561 |
1314 |
|||
18 |
6.361 |
2024 |
|||
19 |
5.417 |
1169 |
|||
20 |
5.461 |
1207 |
( ) : Percentage inhibition
Limit test - test environment
The temperature of the incubator ranged from 21.6 to 25.3°C during the test; although the temperature of the test area slightly exceeded that recommended by the test guidelines (maximum value 25°C), it is not considered to be significant nor to have affected the integrity of the test.
The temperature of the contents of control and test flasks ranged, at the start of the test, from 22.3 to 22.7°C and after 72 hours, from 23.7 to 24.0°C.
The pH of the contents of the flasks measured at the start of the test ranged betwen 7.8 and 8.4; after 72 hours pH ranged from 7.2 to 7.6.
On the day of preparation, the test medium was colourless with particulate material visible on its surface and on the base of the preparation flask. After centrifugation, the medium was colourless with a few particles of material on its surface.
Calculation of results
The effects of test item on the growth of Selenastrum cultures were determined by examining the growth rates of cultures (the rate of change in cell number with time) and biomass (the actual number of cells per volume of medium) as follows. Calculations were based on the the values given in Table 2 above, (quoted to three significant figures) without adjustment for the constant factor of 104.
A = N1– N0 X t1 + N1+ N2– 2N0 X (t2–t1)
2 2
+ Na-1+ Nn– 2N0 X (tn–tn-1)
2
Where:
A = area;
N0= nominal number of cells/ml at t0;
N1= measured number of cells at t1;
N2= measured number of cells at t2;
Nn = measured number of cells at timetn;
T1= time of first measurement after start of test;
t2= time of second measurement after start of test;
tn= time of nthmeasurement after start of test.
The area under the growth curve (biomass) at the test concentration was expressed as a percentage of the solvent control and subtracted from 100 to give the percentage inhibition. The EC50 based on biomass (EbC50) could not be calculated because insufficient inhibition of algal growth was noted.
A statistical comparison of the area under the growth curve in the solvent control and test groups was again carried out using Dunnett’s multicomparison test.
Description of key information
A study was conducted to investigate the effects of test item on the growth of the unicellular green algaSelenastrum capricornutumover a 72 hour period using methods based on OECD Procedure 201 and the Annex to EC Directive 92/69/EEC, Part C3. Exposure of Selenastrum capricornutumto an aqueous mixture of the test substance did not result in a significant reduction of either the average specific growth rates or the mean biomass values compared to solvent controls (p>0.05). Neither the median effect concentration (ErC50 or EbC50) nor the no-observed-effect concentrations could be identified but both must be at least equal to the limit of aqueous solubility of the test material.
Key value for chemical safety assessment
Additional information
Neither the median effect concentration (ErC50 or EbC50) nor the no-observed-effect concentrations could be identified but both must be at least equal to the limit of aqueous solubility of the test material.
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