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EC number: 943-043-9 | CAS number: 8015-91-6
- 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:
- 24/11/2014 - 16/12/2014
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Study according to international guideline (OECD guideline 201) under GLP. No deviations from guideline reported.
Cross-reference
- Reason / purpose for cross-reference:
- reference to same study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 015
- Report date:
- 2015
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
Test material
- Reference substance name:
- Essential oil of Cinnamon bark obtained from the bark of Cinnamomum zeylanicum, Lauraceae by distillation
- Cas Number:
- 8015-91-6
- IUPAC Name:
- Essential oil of Cinnamon bark obtained from the bark of Cinnamomum zeylanicum, Lauraceae by distillation
- Test material form:
- other: Liquid
- Details on test material:
- - Name of test material (as cited in study report): Cinnamon bark oil
- Physical state: liquid
1
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
Not relevant
Sampling and analysis
- Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: nominal Water accomodated fractions (WAF) of 20.0; 8.00; 3.20; 1.28 and 0.512 mg/L.
- Sampling method: The content of total organic carbon (TOC) was measured. Furthermore the total organic carbon in the test item itself was analysed in order to quantify the dissolved content of carbon in the test solution compared to the nominal loading rate. For the calculation of the content of test item in the test solution the blank value of the control was subtracted from the measured value at each loading rate. Analytical samples were taken from each loading rate and control at 0 d (initial value) and after 3 d from aged test solutions. All analytical samples taken were analysed by TOC analysis. These samples were taken from a separate vessel without algae.
Test solutions
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Based on a non-GLP range-finding test the GLP test was performed with the following loading rates (spaced by a geometric factor of 2.5): 20.0; 8.00; 3.20; 1.28 and 0.512 mg/L. Due to low water solubility of the test item two WAFs with nominal loading rates of 20.0 and 8.00 mg/L were prepared. 20.0 mg/L (S1) and 8.00 mg/L (S2) were prepared by weighing the required amount of test item on a weighing scoop and transfer it to a volumetric flask. Test medium was added up to the bench mark. These solutions were stirred by a magnetic stirrer in a closed vessel at room temperature in the dark over 23 hours to dissolve a maximum amount of the test item. Subsequently the undissolved test item was allowed to sediment and/or float for a period of one hour until the phases had separated. The test item solutions were withdrawn from the middle of the preparation vessel by suction (according to OECD Series on Testing and Assessment No. 23). Transfer of undissolved test item was avoided. This was checked by microscopic observation. The test item solutions 3.20, 1.28 and 0.512 mg/L were prepared by dilution of the withdrawn phase of 8.00 mg/L test item stock solution with test medium. Algae cells were added and the solutions were transferred into serum bottles which were filled up to the brim and sealed with crimp caps.
Deviation from protocol: The test item is classified as CMR substance as mentioned in the MSDS. Therefore as a deviation from the original study plan due to technical and security reasons the loading rates 3.20 and 1.28 were not prepared by direct weighting but by dilution of the WAF with 8 mg/l. For safe handling of this CMR substance, all WAF preparations need to be done in a fume cupboard. This has an impact for small amounts of test substances. In addition the maximum solvent volume was reduced to 1 L and 2 L because larger volumes could not be handled in the fume cupboard. Based on these facts the weighing procedure was performed as described in the deviation.
- Controls: yes, blanks
- Evidence of undissolved material (e.g. precipitate, surface film, etc): Transfer of undissolved test item was avoided. This was checked by microscopic observation.
Test organisms
- Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata Hindák
- Strain: SAG 61.81
- Source (laboratory, culture collection): MBM Sciencebridge, Hans-Adolf-Krebs-Weg 1, 37077 Göttingen, Germany.
- Method of cultivation: The algae are grown semi-continuously in sterile liquid cultures in the laboratory. Old medium is periodically replaced by fresh mineral solution in order to keep the algae in an exponential growth state. Stock cultures are also kept on slanted agar tubes and are ordered regularly from the culture collection. Culture conditions were as follows:
- Illumination: continuously (approx. 60 – 120 μE m^-2 s^-1)
- Temperature: 21 – 24 °C
- Culture flasks: 100 mL Erlenmeyer flasks
- CO2 supply: by shaking on a rotating shaker, approximately 105 rpm. Cells from this semi-continuous liquid stock culture will be used for the test.
Study design
- Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- No
Test conditions
- Hardness:
- No data
- Test temperature:
- 21.6 - 23.9°C
- pH:
- 7.40 - 9.33
- Dissolved oxygen:
- Not relevant
- Salinity:
- Not relevant
- Nominal and measured concentrations:
- Nominal (WAF, t=0h): 0.512, 1.28, 3.20, 8.00 and 20.00 mg/L.
Measured (TOC, control corrected, t=0h): 3.30, 6.30, 9.90. 15.9 and 16.9 mg/L
Measured (TOC, control corrected, t=72h): 1.87, 3.78, 6.64, 10.2 and 16.5 mg/L - Details on test conditions:
- TEST SYSTEM
- Test vessel: Serum bottle
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 120 mL serum bottles with crimp caps
- CO2-supply: by shaking on a rotating shaker, 105 rpm
- Initial cells density: 0.55 × 10^4 cells/mL
- Control end cells density: 34.01 x 10^4 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes, AAP medium (Annex 3 of OECD 201)
- Details: 300 mg NaHCO3 were added per 1 L test medium as CO2 supply for the algae in the closed test system.
TEST MEDIUM / WATER PARAMETERS
- Culture medium different from test medium: No
- Intervals of water quality measurement: Measurements of pH-value were performed at t = 0d and t = 3d. The temperature was measured at day 0, 1, 2 and 3 within a separate vessel. The light intensity at 30 different places at test start.
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Photoperiod: continuously
- Light intensity and quality: 4700 – 7100 Lux
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: At defined dates (1, 2 and 3 days), the number of cells in each replicate was determined in duplicate. The determination was performed by means of a calibration curves, where cell numbers (y axis) were plotted versus fluorescence signals (x axis). To establish a calibration curve, the cell numbers were counted with a Neubauer chamber after preparation of a dilution series of a logarithmic growing Pseudokirchneriella subcapitata culture.
The daily fluorescence measurements were performed with a Tecan Reader (infinite 200Pro; serialnumber 1305001441) with an emission wavelength of 670 nm and evaluated with Tecan i-control (Software for Tecan Readers Tecan i-control, 1.10.4.0).
Samples for determination of biomass were taken using a syringe via the septum in the crimp cap. Withdrawn sample volume was replaced with medium which was added via the septum using another syringe.
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5
- Range finding study
- Test concentrations: 0.01, 0.1, 1, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
Results and discussion
Effect concentrationsopen allclose all
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- 8.51 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95%CL: 7.29 - 10.5 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EL10
- Effect conc.:
- 2.91 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95% CL: 2.12 - 3.56 mg/L
- Duration:
- 72 h
- Dose descriptor:
- NOELR
- Effect conc.:
- 3.2 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- LOELR
- Effect conc.:
- 8 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- 4.71 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95% CL: 4.06 - 5.40 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EL10
- Effect conc.:
- 0.945 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95% CL: 0.659 - 1.24 mg/L
- Duration:
- 72 h
- Dose descriptor:
- NOELR
- Effect conc.:
- 8 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- LOELR
- Effect conc.:
- 20 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Details on results:
- - Exponential growth in the control (for algal test): yes
- Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- EC50: ErC50: 0.84 mg/L, ErB50: 0.39 mg/L - Reported statistics and error estimates:
- The statistical evaluation for day 3 was performed for growth rate and yield. The NOEL and LOEL were determined by using a multiple comparison method (Welch Bonferroni-Holms corrected for growth rate and Jonckheere-Terpstra for yield). A test for normality of the data was performed by using the Shapiro-Wilk’s statistic and homogeneity of variance was performed according to Levene. The values for ErL10, ErL50 (growth rate) and EyL10, EyL50 (yield) were determined by probit analysis following the Gompertz distribution. The evaluation of data was performed by SAS® (2002-2010). Negative values were set to zero and values above 100 were set to 100.
Any other information on results incl. tables
Individual cell numbers (x 10^4)
Test Item Loading Rate [mg/L] | Cells/mL) | Yield | |||||||||||||||||||||||||||||||||||||||||
0d | 1d | 2d | 3d | 0d-3d | |||||||||||||||||||||||||||||||||||||||
Control | 0.61 | 1.48 | 5.30 | 27.76 | 27.15 | ||||||||||||||||||||||||||||||||||||||
0.52 | 1.60 | 7.23 | 38.56 | 38.04 | |||||||||||||||||||||||||||||||||||||||
0.58 | 1.53 | 7.27 | 39.13 | 38.55 | |||||||||||||||||||||||||||||||||||||||
0.55 | 1.48 | 6.18 | 34.65 | 34.10 | |||||||||||||||||||||||||||||||||||||||
0.49 | 0.98 | 6.67 | 30.59 | 30.10 | |||||||||||||||||||||||||||||||||||||||
0.57 | 1.32 | 4.65 | 33.37 | 32.80 | |||||||||||||||||||||||||||||||||||||||
Mean | 0.55 | 1.40 | 6.22 | 34.01 | 33.46 | ||||||||||||||||||||||||||||||||||||||
0,512 | 0.67 | 1.36 | 6.02 | 32.12 | 31.45 | ||||||||||||||||||||||||||||||||||||||
0.64 | 1.30 | 7.89 | 34.68 | 34.04 | |||||||||||||||||||||||||||||||||||||||
0.62 | 1.58 | 6.04 | 39.31 | 38.69 | |||||||||||||||||||||||||||||||||||||||
Mean | 0.64 | 1.41 | 6.65 | 35.37 | 34.73 | ||||||||||||||||||||||||||||||||||||||
128 | 0.63 | 1.42 | 6.63 | 26.26 | 25.63 | ||||||||||||||||||||||||||||||||||||||
0.54 | 1.35 | 5.81 | 39.63 | 39.09 | |||||||||||||||||||||||||||||||||||||||
0.61 | 1.51 | 6.28 | -1.57 | -2.18 | |||||||||||||||||||||||||||||||||||||||
Mean | 0.59 | 1.43 | 6.24 | 21.44 | 20.85 | ||||||||||||||||||||||||||||||||||||||
320 | 0.54 | 0.85 | 3.19 | 35.41 | 34.87 | ||||||||||||||||||||||||||||||||||||||
0.61 | 0.85 | 4.25 | 19.20 | 18.59 | |||||||||||||||||||||||||||||||||||||||
0.59 | 0.86 | 4.20 | 33.52 | 32.93 | |||||||||||||||||||||||||||||||||||||||
Mean | 0.58 | 0.85 | 3.88 | 29.38 | 28.80 | ||||||||||||||||||||||||||||||||||||||
800 | 0.53 | 0.67 | 0.77 | 27.00 | 26.47 | ||||||||||||||||||||||||||||||||||||||
0.50 | 0.74 | 0.79 | 2.21 | 1.71 | |||||||||||||||||||||||||||||||||||||||
0.62 | 0.75 | 0.69 | 1.39 | 0.77 | |||||||||||||||||||||||||||||||||||||||
Mean | 0.55 | 0.72 | 0.75 | 10.20 | 9.65 | ||||||||||||||||||||||||||||||||||||||
200 | 0.48 | 0.22 | 0.15 | 1.15 | 0.67 | ||||||||||||||||||||||||||||||||||||||
0.56 | 0.44 | 0.24 | 0.19 | -0.37 | |||||||||||||||||||||||||||||||||||||||
0.48 | 0.29 | 0.12 | 0.13 | -0.35 | |||||||||||||||||||||||||||||||||||||||
Mean | 0.51 | 0.32 | 0.17 | 0.49 | -0.02 |
Control growth rates
Growth rates, µ [d-1] | |||||||||||||||||||||||||||||
0d – 1d | 0d - 2d | 0d - 3d | |||||||||||||||||||||||||||
Control | 0.88634 | 1.08100 |
1.27263 | ||||||||||||||||||||||||||
1.12393 | 131,608 | 143,538 | |||||||||||||||||||||||||||
0.96999 | 1.26424 | 1.40387 | |||||||||||||||||||||||||||
0.98988 | 1.20958 | 1.38104 | |||||||||||||||||||||||||||
0.69315 | 1.30548 | 1.37801 | |||||||||||||||||||||||||||
0.83975 | 1.04949 | 1.35659 | |||||||||||||||||||||||||||
Mean Std. Dev. CV [%] | 0.91717 | 1.20431 | 1.37125 | ||||||||||||||||||||||||||
0.14694 | 0.11447 | 0.05524 | |||||||||||||||||||||||||||
16 | 95 | 40 |
Control daily growth rates
Growth rates, µ [d-1] | |||||||||||||||||||||||||||||
0d – 1d | 1d – 2d | 2d – 3d | |||||||||||||||||||||||||||
Control | 0.88634 | 1.27566 | 1.65589 | ||||||||||||||||||||||||||
1.12393 | 1.50824 | 1.67398 | |||||||||||||||||||||||||||
0.96999 | 1.55849 | 1.68313 | |||||||||||||||||||||||||||
0.98988 | 1.42928 | 1.72398 | |||||||||||||||||||||||||||
0.69315 | 1.91782 | 1.52305 | |||||||||||||||||||||||||||
0.83975 | 1.25924 | 1.97079 | |||||||||||||||||||||||||||
Mean Std. Dev. CV [%] | 0.91717 | 1.49146 | 1.70514 | ||||||||||||||||||||||||||
0.14694 | 0.24121 | 0.14694 | |||||||||||||||||||||||||||
16 | 162 | 86 |
Coefficient of variation (CV) of control daily growth rates
Mean daily growth rates µ [d-1] | Standard deviation | CV [%] | |||||||||||||||||||||||||||||||||||
1.27263 | 0.38478 | 30 | |||||||||||||||||||||||||||||||||||
1.43538 | 0.28217 | 20 | |||||||||||||||||||||||||||||||||||
1.40387 | 0.38088 | 27 | |||||||||||||||||||||||||||||||||||
1.38105 | 0.36942 | 27 | |||||||||||||||||||||||||||||||||||
1.37801 | 0.62509 | 45 | |||||||||||||||||||||||||||||||||||
1.35659 | 0.57177 | 42 | |||||||||||||||||||||||||||||||||||
Mean CV | 32 |
Applicant's summary and conclusion
- Validity criteria fulfilled:
- yes
- Remarks:
- Control cell number increase and coefficients of variation meet validity criteria
- Conclusions:
- The acute toxicity (72h-ErL50) of Cinnamon bark oil towards Pseudokirchneriella subcapitata is 8.51 mg/L.
- Executive summary:
The acute toxicity of Cinnamon bark oil towards Pseudokirchneriella subcapitata was investigated according to OECD guideline 201 under GLP. Algal cells were exposed to nominal WAFs of 0.512, 1.28, 3.2, 8 and 20 mg/L and were observed for 72 hours. Based on the nominal loading rates the 72h-ErL50 and 72h-ErL10 were found to 8.51 and 2.91 mg/L respectively.
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