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EC number: 700-774-6 | CAS number: -
Toxicity to aquatic algae and cyanobacteria
Administrative data
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- May to October 2011
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: the study was performed according to OECD guideline and GLP
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 011
- Report date:
- 2011
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- yes
- Remarks:
- The concentration of NaHCO3 was increased to be twice of that indicated in the Guideline 201. This concentration has been found to be optimal.
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Absorbent N grade A
- IUPAC Name:
- Absorbent N grade A
- Details on test material:
- - Name of test material (as cited in study report): Absorbent N grade A
- Substance type: UVCB
- Physical state: Brown liquid
- Stability under test conditions: months
- Storage condition of test material: room temperature, in the dark
Constituent 1
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: 1, 2, 4, 8, 10 mg/l
- Sampling method: data not available
- Sample storage conditions before analysis: data not available
Test solutions
- Vehicle:
- no
- Details on test solutions:
- The test item was introduced into the test solutions using aqueous extracts („WAFs“) from the test item as follows: First, suspensions of the test item prepared in ultrapure water were shaken on a shaking machine for 24 h at ≈130 rpm at room temperature, respectively. During that time an equilibration between the test item and the water was considered to be achieved. After 24h, shaking was stopped, and the suspensions were filtered through glass fibre filter. The eluates („WAFs“) were used directly without any further dilution steps. The eluates were used for the test. By the addition of the "intermediate dilution" and the algal inoculation the final nominal concen¬trations (loadings, see below) were achieved. For the inoculated test solutions with the test item, as well as those without any test item, 7 parallels (main test; 5 parallels within the screening) were incubated under constant light. As reagents’ Blank without algal inoculation three test solutions were prepared as described above containing a 1:10 diluted inter¬mediate dilution instead of the algal pre-culture. These blanks were incubated under the same conditions under constant light as the test solution to be tested. The measured turbidity in those test solutions was subtracted from those with the algal inoculation. In the same way, seven control solutions without any test item but with algal inoculation were prepared within the main test as well as one test solution without any test item and without algal inoculation.
Test organisms
- Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- TEST ORGANISM
- Common name: green alga
- Strain: Desmodesmus subspicatus CHODAT
- Source (laboratory, culture collection): Sammlung von Algenkulturen, Pflanzen¬physiologi¬sches Institut der Universität Göttingen [SAG], Strain-No. 86.81
- Age of inoculum (at test initiation): 3 days
- Method of cultivation: The algae are cultivated under aseptic conditions.
ACCLIMATION
- Acclimation period:
- Culturing media and conditions (same as test or not): same as test
- Any deformed or abnormal cells observed: data not available
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- none
Test conditions
- Hardness:
- data not available
- Test temperature:
- The temperature was within a range of 23.8 to 24.6°C
- pH:
- Test Solution / Concentration Test Item pH at t0 pH at t72h
Control with algal inoculation 8.34 10.75
Control without algal inoculation 8.33 8.27
1 mg/L with algal inoculation 8.32 10.59
1 mg/L without algal inoculation 8.32 8.17
2 mg/L with algal inoculation 8.31 9.65
2 mg/L without algal inoculation 8.31 8.18
4 mg/L with algal inoculation 8.31 8.48
4 mg/L without algal inoculation 8.32 8.17
8 mg/L with algal inoculation 8.32 8.40
8 mg/L without algal inoculation 8.31 8.23
10 mg/L with algal inoculation 8.32 8.34
10 mg/L without algal inoculation 8.31 8.25 - Dissolved oxygen:
- data not available
- Salinity:
- not applicable (freshwater)
- Nominal and measured concentrations:
- Mean Concentrations in the Treatment over the whole Incubation Time [geometric Mean]
Nominal Conc. Mean Measured Concentrations (mg/L)
1 1.25
2 1.73
4 4.28
8 8.33
10 11.17 - Details on test conditions:
- TEST SYSTEM
- Test vessel: 50 mL-glass-flasks with crimp-on caps, heat sterilized
- Material, size, headspace, fill volume: filled with 50 mL
- Aeration: In order to assure the input of CO2, the test solutions were stirred by means of a magnetic stirrer for the duration of 15 minutes per hour
- Initial cells density: 2.64*10E3 cells/mL
- Control end cells density: 7.58*10E5 cells/mL
- No. of vessels per concentration (replicates): 7
- No. of vessels per control (replicates): 7
GROWTH MEDIUM
- Standard medium used: yes
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: 24/24h under constant light
- Light intensity and quality: illumination rate of >=120 μE/m2s ([~8000 Lux]
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: The increase in biomass was determined using a spectrophotometer after 22.5 h, 46.75 h and 72 h (main test). As a measure for the biomass the absorbance of the test solutions measured at 578 nm was used.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study
- Test concentrations: 10; 100; 1000 mg/L
- Results used to determine the conditions for the definitive study: At all three test concentrations no algae growth was observed. - Reference substance (positive control):
- not specified
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 1.202 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC20
- Effect conc.:
- 1.993 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- 5.246 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- <= 1 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- < 1 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): data not available - Results with reference substance (positive control):
- not applicable
- Reported statistics and error estimates:
- not applicable
Any other information on results incl. tables
Cell Numbers
Time t0 |
Control |
Nominal Concentrations |
|||||
1 mg/L |
2 mg/L |
4 mg/L |
8 mg/L |
10 mg/L |
|||
|
|
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
|
|
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
|
|
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
|
|
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
|
|
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
|
|
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
|
|
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
Mean Values: |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
2.64*103 |
|
Time t26.25h |
Control |
Nominal Concentrations |
|||||
1 mg/L |
2 mg/L |
4 mg/L |
8 mg/L |
10 mg/L |
|||
|
|
6.08*104 |
2.08*104 |
1.24*104 |
2.33*103 |
1.73*103 |
5.41*103 |
|
|
3.46*104 |
2.47*104 |
1.24*104 |
4.17*103 |
1.73*103 |
5.41*103 |
|
|
3.07*104 |
3.07*104 |
1.05*104 |
4.17*103 |
1.73*103 |
3.55*103 |
|
|
3.07*104 |
1.88*104 |
1.24*104 |
4.17*103 |
1.73*103 |
5.41*103 |
|
|
2.87*104 |
2.67*104 |
8.55*103 |
2.33*103 |
1.73*103 |
3.55*103 |
|
|
4.26*104 |
2.67*104 |
1.24*104 |
6.03*103 |
1.73*103 |
3.55*103 |
|
|
2.67*104 |
1.88*104 |
8.55*103 |
6.03*103 |
1.73*103 |
5.41*103 |
Mean Values: |
3.64*104 |
2.39*104 |
1.10*104 |
4.18*103 |
1.73*103 |
4.61*103 |
|
Time t50.75h |
Control |
Nominal Concentrations |
|||||
1 mg/L |
2 mg/L |
4 mg/L |
8 mg/L |
10 mg/L |
|||
|
|
2.26*105 |
1.48*105 |
7.31*104 |
2.01*104 |
6.03*103 |
2.94*10 |
|
|
2.09*105 |
1.46*105 |
7.72*104 |
1.43*104 |
6.03*103 |
2.94*103 |
|
|
1.98*105 |
1.39*105 |
7.51*104 |
1.62*104 |
6.03*103 |
4.79*103 |
|
|
1.98*105 |
1.48*105 |
6.49*104 |
1.43*104 |
2.33*103 |
4.79*103 |
|
|
1.96*105 |
1.42*105 |
7.51*104 |
1.62*104 |
4.17*103 |
4.79*103 |
|
|
2.01*105 |
1.42*105 |
6.70*104 |
1.43*104 |
2.33*103 |
2.94*103 |
|
|
1.94*105 |
1.50*105 |
6.49*104 |
2.80*104 |
2.33*103 |
2.94*103 |
Mean Values: |
2.03*105 |
1.45*105 |
7.10*104 |
1.77*104 |
4.18*103 |
3.73*103 |
|
Time t72h |
Control |
Nominal Concentrations |
|||||
1 mg/L |
2 mg/L |
4 mg/L |
8 mg/L |
10 mg/L |
|||
|
|
7.12*105 |
5.76*105 |
2.88*105 |
6.15*104 |
1.56*104 |
1.69*104 |
|
|
7.37*105 |
5.43*105 |
3.03*105 |
5.14*104 |
1.95*104 |
2.67*104 |
|
|
7.46*105 |
5.56*105 |
3.10*105 |
5.54*104 |
1.56*104 |
2.87*104 |
|
|
7.59*105 |
6.05*105 |
2.77*105 |
6.15*104 |
1.56*104 |
1.49*104 |
|
|
7.72*105 |
5.65*105 |
3.31*105 |
6.15*104 |
1.37*104 |
2.08*104 |
|
|
7.86*105 |
5.78*105 |
3.01*105 |
4.13*104 |
1.75*104 |
1.88*104 |
|
|
7.92*105 |
6.20*105 |
2.86*105 |
4.13*104 |
2.14*104 |
1.30*104 |
Mean Values: |
7.58*105 |
5.78*105 |
2.99*105 |
5.34*104 |
1.70*104 |
2.00*104 |
|
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Remarks:
- The test is considered valid as all conditions for validity were met
- Conclusions:
- On the Basis of the nominal Concentrations tested (in mg/l)
Yield LOEC ≤ 1.000
NOEC < 1.000
Growth rate LOEC ≤ 1.000
NOEC < 1.000 - Executive summary:
The toxicity of“Absorbent N grade A“towards algae was testedaccording to OECD-Guideline No. 201, in the Version dated 23-March 2006. The toxic effect was investigated by determination of the inhibition of the growth rate of the algae and the yield during the exposure period of 72 h. As the test item is partially poorly water-soluble, it was tested using aqueous extracts (“WAFs”) from up to 1000 mg test item/L (final concentration). The main test was performed using “WAFs” from 1 to 10 mg/L of the test item. As a conclusion of the analytical part of this study, recoveries of the concentrations determined after 72h were all at ≥ 80% of those determined at time t0, and furthermore,the measured concentrations of the treatments did not decrease more than 20 % from the nominal values. Based on this, the reported effect concentrations refer to the nominal concentrations tested.From the results of the main test, toxic effects were determined with the following values:
On the Basis of the nominal Concentrations tested
Yield (0 - 72 h)
EC10 0.697
95%-CL lower 0.657
upper 0.735
EC20 0.933
95%-CL lower 0.893
upper 0.971
EC50 1.631
95%-CL lower 1.590
upper 1.672
Yield LOEC ≤ 1.000
NOEC < 1.000
Growth rate (0 - 72 h)
EC10 1.202
95%-CL lower 0.993
upper 1.405
EC20 1.993
95%-CL lower 1.740
upper 2.233
EC50 5.246
95%-CL lower 4.889
upper 5.633
Growth rate LOEC ≤ 1.000
NOEC < 1.000
The test is considered valid as all conditions for validity were met.
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