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Diss Factsheets

Ecotoxicological information

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:
2011
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

Constituent 1
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

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.