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EC number: 218-254-8 | CAS number: 2094-98-6
- 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
- Endpoint:
- toxicity to aquatic algae and cyanobacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 03.-07.12.18
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
- Version / remarks:
- 2006
- Deviations:
- yes
- Remarks:
- uncritical: stock solutions were autoclaved for 20 minutes, pre-culture was prepared 5 days before test start (validity criteria were observed)
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- 1,1'-azobis(1-cyclohexanecarbonitrile)
- EC Number:
- 218-254-8
- EC Name:
- 1,1'-azobis(1-cyclohexanecarbonitrile)
- Cas Number:
- 2094-98-6
- Molecular formula:
- C14H20N4
- IUPAC Name:
- 1,1'-azobis(1-cyclohexanecarbonitrile)
- Test material form:
- solid: granular
Constituent 1
Sampling and analysis
- Analytical monitoring:
- yes
Test solutions
- Vehicle:
- no
Test organisms
- Test organisms (species):
- Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)
- Details on test organisms:
- TEST ORGANISM
- Common name: Green Algae - Chlorophytes
- Strain: 86.81
- Source (laboratory, culture collection): MBM Sciencebridge GmbH (Institut für Pflanzenphysiologie of Universität Göttingen), Jan 2006
- Age of inoculum (at test initiation):
- Method of cultivation:
ACCLIMATION
- Acclimation period:
- Culturing media and conditions (same as test or not):
- Any deformed or abnormal cells observed:
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
Test conditions
- Test temperature:
- 21.7 – 22.8 °C
- pH:
- 7.6-8.3
- Nominal and measured concentrations:
- nominal concentration: 1 / 3.2 / 10 / 32 / 100 mg/L
geometric mean: 0.03 / 0.06 / 0.21 / 0.71 / 1.98 mg/L - Details on test conditions:
- TEST SYSTEM
- Test vessel: glass flasks
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 45 ±1 mL
- Aeration:
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- Initial cells density: 2.4E3
- Control end cells density: 4.2E5
- No. of organisms per vessel:
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
- No. of vessels per vehicle control (replicates): -
GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used:
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Conductivity:
- Culture medium different from test medium:
- Intervals of water quality measurement:
OTHER TEST CONDITIONS
- Sterile test conditions: yes/no
- Adjustment of pH:
- Photoperiod:
- Light intensity and quality: 5000 lux
- Salinity (for marine algae):
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: electronic particle counter (Beckman Coulter Z2)
- Chlorophyll measurement: -
- Other: -
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.2
- Justification for using less concentrations than requested by guideline: -
- Range finding study
- Test concentrations: not stated (non-GLP pre-test)
- Results used to determine the conditions for the definitive study: yes - Reference substance (positive control):
- yes
- Remarks:
- K2Cr2O7 (CAS No. 7778-50-9)
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- NOEC
- Effect conc.:
- 0.02 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOEC
- Effect conc.:
- 0.06 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC10
- Effect conc.:
- 0.95 mg/L
- Nominal / measured:
- meas. (geom. mean)
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Key result
- Duration:
- 72 h
- Dose descriptor:
- EC50
- Effect conc.:
- > 1.95 mg/L
- Nominal / measured:
- meas. (geom. mean)
- 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): no
- Unusual cell shape: no
- Colour differences: no
- Flocculation: no
- Adherence to test vessels: no
- Aggregation of algal cells: no
- Other:
- Any stimulation of growth found in any treatment: yes (@ nominal concentration: 1 mg/L)
- 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):
- 72h EC50 0.79 mg/L
Any other information on results incl. tables
calculated cell numbers
nominal concentration mg/L | replicate | cell number/mL | |||
0 h | 24 h | 48 h | 72 h | ||
Blank control | 1 | 2380 | 8200 | 33120 | 379600 |
2 | 2380 | 5380 | 36020 | 420680 | |
3 | 2380 | 8120 | 45520 | 380580 | |
4 | 2380 | 9220 | 55620 | 414420 | |
5 | 2380 | 9980 | 68280 | 459980 | |
6 | 2380 | 9700 | 70740 | 459360 | |
mean | 2380 | 8433 | 51550 | 419103 | |
SD | 0 | 1679 | 16012 | 35666 | |
1 | 1 | 2380 | 10600 | 77440 | 492680 |
2 | 2380 | 10680 | 73500 | 477380 | |
3 | 2380 | 9580 | 66620 | 454700 | |
mean | 2380 | 10287 | 72520 | 474920 | |
SD | 0 | 613 | 5476 | 19109 | |
3.2 | 1 | 2380 | 9380 | 48140 | 399620 |
2 | 2380 | 8300 | 41340 | 324500 | |
3 | 2380 | 8460 | 33400 | 372520 | |
mean | 2380 | 8713 | 40960 | 365547 | |
SD | 0 | 583 | 7377 | 38042 | |
10 | 1 | 2380 | 5120 | 29980 | 338340 |
2 | 2380 | 8960 | 38140 | 332040 | |
3 | 2380 | 10760 | 58280 | 342060 | |
mean | 2380 | 8280 | 42133 | 337480 | |
SD | 0 | 2881 | 14566 | 5065 | |
32 | 1 | 2380 | 11360 | 60860 | 330560 |
2 | 2380 | 12180 | 73140 | 348840 | |
3 | 2380 | 12400 | 76740 | 362880 | |
mean | 2380 | 11980 | 70247 | 347427 | |
SD | 0 | 548 | 8326 | 16206 | |
100 | 1 | 2380 | 11260 | 34640 | 37140 |
2 | 2380 | 10720 | 42400 | 39740 | |
3 | 2380 | 10360 | 35320 | 35620 | |
mean | 2380 | 10780 | 37453 | 37500 | |
SD | 0 | 453 | 4297 | 2083 |
growth rate
nominal conc. in mg/L | replicate | growth rate [day-1] | ||
24 h | 48 h | 72 h | ||
Blank control | 1 | 1.24 | 1.32 | 1.69 |
2 | 0.82 | 1.36 | 1.72 | |
3 | 1.23 | 1.48 | 1.69 | |
4 | 1.35 | 1.58 | 1.72 | |
5 | 1.43 | 1.68 | 1.75 | |
6 | 1.41 | 1.70 | 1.75 | |
mean | 1.25 | 1.52 | 1.72 | |
SD | 0.23 | 0.16 | 0.03 | |
1 | 1 | 1.49 | 1.74 | 1.78 |
2 | 1.50 | 1.72 | 1.77 | |
3 | 1.39 | 1.67 | 1.75 | |
mean | 1.46 | 1.71 | 1.77 | |
SD | 0.06 | 0.04 | 0.01 | |
3.2 | 1 | 1.37 | 1.50 | 1.71 |
2 | 1.25 | 1.43 | 1.64 | |
3 | 1.27 | 1.32 | 1.68 | |
mean | 1.30 | 1.42 | 1.68 | |
SD | 0.07 | 0.09 | 0.04 | |
10 | 1 | 0.77 | 1.27 | 1.65 |
2 | 1.33 | 1.39 | 1.65 | |
3 | 1.51 | 1.60 | 1.66 | |
mean | 1.20 | 1.42 | 1.65 | |
SD | 0.39 | 0.17 | 0.01 | |
32 | 1 | 1.56 | 1.62 | 1.64 |
2 | 1.63 | 1.71 | 1.66 | |
3 | 1.65 | 1.74 | 1.68 | |
mean | 1.62 | 1.69 | 1.66 | |
SD | 0.05 | 0.06 | 0.02 | |
100 | 1 | 1.55 | 1.34 | 0.92 |
2 | 1.51 | 1.44 | 0.94 | |
3 | 1.47 | 1.35 | 0.90 | |
mean | 1.51 | 1.38 | 0.92 | |
SD | 0.04 | 0.06 | 0.02 |
daily growth rates
growth rates | days 0-1 | days 1-2 | days 2-3 | CV of sectional daily growth rates | days 0-3 |
Replicate 1 | 1.237 | 1.396 | 2.439 | 39 % | 1.691 |
Replicate 2 | 0.816 | 1.901 | 2.458 | 48 % | 1.725 |
Replicate 3 | 1.227 | 1.724 | 2.124 | 27 % | 1.692 |
Replicate 4 | 1.354 | 1.797 | 2.008 | 19 % | 1.720 |
Replicate 5 | 1.433 | 1.923 | 1.908 | 16 % | 1.755 |
Replicate 6 | 1.402 | 1.987 | 1.871 | 18 % | 1.754 |
mean | 1.245 | 1.788 | 2.135 | 28 % | 1.723 |
SD | 0.277 | 0.214 | 0.259 | 0.028 | |
CV | 18 % | 12 % | 12 % | 2 % |
Validity criteria
Parameter | Validity criteria | observed value | assessment |
increase factor biomass | factor 16 in 72 h | 176 | valid |
mean coefficient of variation of daily growth rates | max 35 % | 28 % | valid |
Coefficient of variation of average growth rate during the whole test period | max 7 % | 2 % | valid |
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Executive summary:
The study was performed using 5 concentrations ranging from 1 to 100 mg/L (nominal). Incubation time (test system Desmodesmus subspicatus)was 72 hours. The cell concentration of each replicate was determined by measuring the cell numbers every 24 hours with an electronic particle counter. Growth rate µ and the yield were determined from the cell number at the respective observation times.
Significant inhibition of algal growth was observed at the following concentrations:
3.2 – 100 mg/L (nominal concentrations)
At the beginning and at the end of the test, the content of the test item in the test solutions was determined using HPLC-UV-determination. Because of the poor solubility of the test item the measured concentrations lay between 1 % and 4 % of the nominal concentration during the test. Therefore, the determination of the results was based on the geometric mean of the measured concentrations.
The 72h-EC50values of potassium dichromate (K2Cr2O7, CAS No. 7778-50-9) were determined in a separate reference test. The values lay within the range of the laboratory (growth rate 0.73 - 1.10 mg/L, yield 0.21 – 0.66 mg/L).
The following results for the test item 1,1'-Azobis(cyclohexane-1-carbonitrile) were determined:
Table 3‑a Results of the test item
Endpoint
NOEC
LOEC
EC10
EC50
Growth Rate
0.02 mg/L
0.06 mg/L
0.95 mg/L
> 1.85 mg/L
Yield
0.02 mg/L
0.06 mg/L
0.34 mg/L
0.94 mg/L
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