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EC number: 286-249-8 | CAS number: 85203-56-1 Extractives and their physically modified derivatives such as tinctures, concretes, absolutes, essential oils, oleoresins, terpenes, terpene-free fractions, distillates, residues, etc., obtained from Eucalyptus maculata citriodora, Myrtaceae.
- 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:
- 14/1/2013 - 16/1/2013
- 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.
- Reason / purpose for cross-reference:
- reference to same study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 201 (Alga, Growth Inhibition Test)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method C.3 (Algal Inhibition test)
- Deviations:
- no
- Principles of method if other than guideline:
- Not relevant
- GLP compliance:
- yes (incl. QA statement)
- Specific details on test material used for the study:
- Details on properties of test surrogate or analogue material (migrated information):
Not relevant - Analytical monitoring:
- yes
- Details on sampling:
- - Concentrations: 6.25, 12.5, 25, 50 and 100 mg/L
- Sampling method: Samples were taken from the uninoculated control and the 6.25, 12.5, 25, 50 and 100 mg/L loading rate WAF test groups at 0 and 72 hours for this analysis.
- Sample storage conditions before analysis: stored at approximately -20ºC for further analysis if necessary. - Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Amounts of test item (72.5, 70, 140, 120 and 230 mg) were each separately added to the surface of 11.6, 5.6, 5.6, 2.4 and 2.3 liters of culture medium to give the 6.25, 12.5, 25, 50 and 100 mg/L loading rates respectively. After the addition of the test item, the culture medium was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 47 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aqueous phase or WAF was removed by mid-depth siphoning (the first 75-100 mL discarded) to give the 6.25, 12.5, 25, 50 and 100 mg/L loading rate WAFs. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The test item was weighed out under non-actinic light, and the preparations were shielded from light during the mixing period due to the light-sensitive nature of the test item. An aliquot (1 liter) of each of the loading rate WAFs was separately inoculated with algal suspension (7.4 mL) to give the required test concentrations of 6.25, 12.5, 25, 50 and 100 mg/L loading rate WAF.
- Controls: Yes, blanks
- Evidence of undissolved material (e.g. precipitate, surface film, etc): At both the start and end of the mixing period, and following a 1-hour standing period, the 6.25 and 12.5 mg/L loading rate WAFs were observed to be clear colorless water columns. The 25, 50 and 100 mg/L loading rate WAFs were observed to be clear colorless water columns with globules of test item on the surface. Microscopic observations of the WAFs after siphoning showed no microscopic particles to be present. At the start of the test all control and test cultures were observed to be clear colorless solutions. After the 72-Hour test period all control and the 6.25 and 12.5 mg/L loading rate WAF test preparations were observed to be pale green dispersions. The 25, 50 and 100 mg/L loading rate WAF test preparations were observed to be clear colorless solutions. - Test organisms (species):
- Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
- Details on test organisms:
- TEST ORGANISM
- Common name: Pseudokirchnerella subcapitata
- Strain: CCAP 278/4
- Source (laboratory, culture collection): Culture Collection of Algae and Protozoa (CCAP), SAMS Research Services Ltd, Scottish Marine Institute, Oban, Argyll, Scotland. Master cultures were maintained in the laboratory by the periodic replenishment of culture medium.
- Method of cultivation: The master cultures were maintained in the laboratory under constant aeration and illumination at 21 ± 1ºC.
ACCLIMATION
- Culturing media and conditions (same as test or not): Prior to the start of the test sufficient master culture was added to approximately 100 mL volumes of culture media contained in conical flasks to give an initial cell density of approximately 103 cells/mL. The flasks were plugged with polyurethane foam stoppers and kept under constant agitation by orbital shaker (100 – 150 rpm) and constant illumination at 24 ± 1ºC until the algal cell density was approximately 10^4 – 10^5 cells/mL. - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 72 h
- Post exposure observation period:
- Not relevant
- Hardness:
- No data
- Test temperature:
- 24 +/- 1ºC
- pH:
- 7.3 - 9.6 (maximum increase of 1.4 during test; maximum decrease of 0.5 during test)
- Dissolved oxygen:
- Not relevant
- Salinity:
- Not relevant
- Nominal and measured concentrations:
- Nominal: 6.25, 12.5, 25, 50 and 100 mg/L
Measured (t=0h):Measured (t=72h): 1.55, 9.36, 3.57, 17.83 and 18.07 mg/L - Details on test conditions:
- TEST SYSTEM
- Test vessel: conical flask
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: glass, 300 mL volume filled with 300 mL test medium
- Initial cells density: 5.3*10^3 cell/mL
- Control end cells density: 3.36x10^5
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes
- Detailed composition if non-standard medium was used:
NaNO3 25.5 mg/L
MgCl2.6H2O 12.164 mg/L
CaCl2.2H2O 4.41 mg/L
MgSO4.7H2O 14.7 mg/L
K2HPO4 1.044 mg/L
NaHCO3 15.0 mg/L
H3BO3 0.1855 mg/L
MnCl2.4H2O 0.415 mg/L
ZnCl2 0.00327 mg/L
FeCl3.6H2O 0.159 mg/L
CoCl2.6H2O 0.00143 mg/L
Na2MoO4.2H2O 0.00726 mg/L
CuCl2.2H2O 0.000012 mg/L
Na2EDTA.2H2O 0.30 mg/L
The culture medium was prepared using reverse osmosis purified deionized water* and the pH adjusted to 7.5 ± 0.1 with 0.1N NaOH or HCl.
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reverse osmosis purified deionized water*
- Culture medium different from test medium: No
- Intervals of water quality measurement: The pH of the control and each test preparation was determined at initiation of the test and after 72 hours exposure. The pH was measured using a WTW pH 320 pH meter. The temperature within the incubator was recorded daily.
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: pH adjusted to 7.5 ± 0.1 with 0.1N NaOH or HCl.
- Photoperiod: continuous
- Light intensity and quality: warm white lighting (380 – 730 nm) at 7000 lux
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: electronic particle counter
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study
- Test concentrations:10 and 100 mg/L - Reference substance (positive control):
- yes
- Remarks:
- Potassium dichromate
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- 18 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Remarks on result:
- other: 95%CI: 17 - 20 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EL10
- Effect conc.:
- 12 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- NOELR
- Effect conc.:
- 12.5 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- growth rate
- Duration:
- 72 h
- Dose descriptor:
- EL50
- Effect conc.:
- 13 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Remarks on result:
- other: 95%CI: 13 - 13 mg/L
- Duration:
- 72 h
- Dose descriptor:
- EL10
- Effect conc.:
- 11 mg/L
- Nominal / measured:
- nominal
- Conc. based on:
- test mat.
- Basis for effect:
- biomass
- Duration:
- 72 h
- Dose descriptor:
- NOELR
- Effect conc.:
- 12.5 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
- Observation of abnormalities (for algal test): All test and control cultures were inspected microscopically at 72 hours. After 72 hours there were no abnormalities detected in the control or test cultures at 6.25 loading rate WAF, however cell debris was observed to be present in the test cultures at 12.5 and 25 mg/L loading rate WAF and few to no cells were observed at 50 and 100 mg/L loading rate WAF.
- Any stimulation of growth found in any treatment: yes, in two of the 6.25 mg/L vessels.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: At both the start and end of the mixing period, and following a 1-hour standing period, the 6.25 and 12.5 mg/L loading rate WAFs were observed to be clear colorless water columns. The 25, 50 and 100 mg/L loading rate WAFs were observed to be clear colorless water columns with globules of test item on the surface. Microscopic observations of the WAFs after siphoning showed no microscopic particles to be present. At the start of the test all control and test cultures were observed to be clear colorless solutions. After the 72-Hour test period all control and the 6.25 and 12.5 mg/L loading rate WAF test preparations were observed to be pale green dispersions. The 25, 50 and 100 mg/L loading rate WAF test preparations were observed to be clear colorless solutions. - Results with reference substance (positive control):
- - Results with reference substance valid? Yes
- ErC50: 1.2 mg/L
- EbC50: 0.52 mg/L - Reported statistics and error estimates:
- One way analysis of variance incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf 1981) and Dunnett's multiple comparison procedure for comparing several treatments with a control (Dunnett 1955) was carried out on the growth rate and yield data after 72 hours for the control and all test loading rates to determine any statistically significant differences between the test and control groups. All statistical analyses were performed using the SAS computer software package (SAS 1999 - 2001).
- Validity criteria fulfilled:
- yes
- Remarks:
- > 16-fold increase in controls, mean of coefficients of variation < 35%, specific coefficient of variation < 7%
- Conclusions:
- The ecotoxicity (72h-ErL10 and 72h-ErL50) of Eucalyptus citriodora towards Pseudokirchnerella subcapitata is 12 and 18 mg/L respectively.
- Executive summary:
The ecotoxicity of Eucalyptus citriodora towards Pseudokirchnerella subcapitata was investigated according to OECD guideline 201 under GLP. Algal cells were exposed to WAFs with nominal loading rates of 6.25, 12.5, 25, 50 and 100 mg/L and were observed for 72 hours. The 72h-ErL10 and 72h-ErL50 were found to be 12 and 18 mg/L respectively based on nominal concentrations.
Reference
Cell densities and pH-values:
Nominal Loading Rate (mg/L) |
pH |
Cell Densities*(cells per mL) |
pH |
||||
0 h |
0 h |
24 h |
46 h |
72 h |
72 h |
||
Control
|
R1 |
8.6 |
2.91E+03 |
2.33E+04 |
1.04E+05 |
4.56E+05 |
9.6 |
R2 |
2.54E+03 |
2.40E+04 |
1.14E+05 |
4.74E+05 |
|||
R3 |
3.89E+03 |
2.61E+04 |
7.55E+04 |
2.56E+05 |
|||
R4 |
3.46E+03 |
2.41E+04 |
1.22E+05 |
4.85E+05 |
|||
R5 |
4.21E+03 |
2.21E+04 |
9.06E+04 |
3.66E+05 |
|||
R6 |
5.71E+03 |
2.47E+04 |
9.28E+04 |
3.57E+05 |
|||
Mean |
3.79E+03 |
2.41E+04 |
9.97E+04 |
3.99E+05 |
|||
6.25
|
R1 |
8.1 |
2.48E+03 |
2.35E+04 |
1.18E+05 |
1.13E+06 |
9.5 |
R2 |
2.64E+03 |
2.04E+04 |
1.01E+05 |
3.61E+05 |
|||
R3 |
2.37E+03 |
1.93E+04 |
8.15E+04 |
3.24E+05 |
|||
Mean |
2.50E+03 |
2.10E+04 |
1.02E+05 |
6.04E+05 |
|||
12.5
|
R1 |
8.0 |
2.53E+03 |
1.88E+04 |
4.74E+04 |
1.36E+05 |
9.1 |
R2 |
3.23E+03 |
1.82E+04 |
6.79E+04 |
3.32E+05 |
|||
R3 |
5.12E+03 |
1.87E+04 |
6.52E+04 |
53.10E+05 |
|||
Mean |
3.63E+03 |
1.85E+04 |
6.02E+04 |
2.59E+05 |
|||
25
|
R1 |
8.0 |
3.94E+03 |
1.16E+04 |
1.17E+04 |
1.90E+04 |
7.9 |
R2 |
2.34E+03 |
6.19E+03 |
6.24E+03 |
7.23E+03 |
|||
R3 |
5.01E+03 |
7.17E+03 |
5.70E+03 |
7.44E+03 |
|||
Mean |
3.76E+03 |
8.33E+03 |
7.87E+03 |
1.12E+04 |
|||
50
|
R1 |
8.0 |
3.30E+03 |
9.57E+03 |
1.06E+04 |
9.76E+03 |
7.9 |
R2 |
2.96E+03 |
5.81E+03 |
7.17E+03 |
3.42E+03 |
|||
R3 |
4.11E+03 |
6.08E+03 |
7.74E+03 |
4.08E+03 |
|||
Mean |
3.46E+03 |
7.15E+03 |
8.49E+03 |
5.75E+03 |
|||
100
|
R1 |
7.8 |
5.79E+03 |
8.59E+03 |
6.13E+03 |
4.82E+03 |
7.3 |
R2 |
4.10E+03 |
7.97E+03 |
6.70E+03 |
5.04E+03 |
|||
R3 |
3.58E+03 |
7.42E+03 |
6.24E+03 |
5.14E+03 |
|||
Mean |
4.49E+03 |
7.99E+03 |
6.36E+03 |
5.00E+03 |
R1- R6= Replicates 1 to 6
Daily specific growth rates in controls:
Daily Specific Growth Rate (cells/mL/hour) |
||||
Day 0 - 1 |
Day 1 - 2 |
Day 2 - 3 |
||
Control |
R1 |
0.064 |
0.062 |
0.062 |
R2 |
0.065 |
0.065 |
0.059 |
|
R3 |
0.069 |
0.044 |
0.051 |
|
R4 |
0.066 |
0.067 |
0.058 |
|
R5 |
0.062 |
0.059 |
0.058 |
|
R6 |
0.067 |
0.055 |
0.056 |
|
Mean |
0.066 |
0.059 |
0.057 |
Nominal Loading Rate |
Growth Rate (cells/mL/hour) |
Yield (cells/mL) |
|||
0 – 72 h |
% Inhibition |
0 – 72 h |
% Inhibition* |
||
Control
|
R1 |
0.063 |
|
4.53E+05 |
|
R2 |
0.063 |
|
4.71E+05 |
|
|
R3 |
0.055 |
|
2.52E+05 |
|
|
R4 |
0.064 |
- |
4.81E+05 |
- |
|
R5 |
0.060 |
|
3.61E+05 |
|
|
R6 |
0.059 |
|
3.52E+05 |
|
|
Mean |
0.061 |
|
3.95E+05 |
|
|
SD |
0.003 |
|
8.94E+04 |
|
|
1.0
|
R1 |
0.075 |
[23] |
1.12E+06 |
|
R2 |
0.059 |
3 |
3.58E+05 |
|
|
R3 |
0.058 |
5 |
3.22E+05 |
|
|
Mean |
0.064 |
[5] |
6.01E+05 |
[52] |
|
SD |
0.010 |
|
4.52E+05 |
|
|
3.2
|
R1 |
0.046 |
25 |
1.34E+05 |
|
R2 |
0.058 |
5 |
3.29E+05 |
|
|
R3 |
0.057 |
7 |
3.05E+05 |
|
|
Mean |
0.054 |
12 |
2.56E+05 |
35 |
|
SD |
0.007 |
|
1.06E+05 |
|
|
10
|
R1 |
0.019 |
69 |
1.51E+04 |
|
R2 |
0.005 |
92 |
4.90E+03 |
|
|
R3 |
0.006 |
90 |
2.43E+03 |
|
|
Mean |
0.010 |
84 |
7.47E+03 |
98 |
|
SD |
0.008 |
|
6.71E+03 |
|
|
32
|
R1 |
0.009 |
85 |
6.46E+03 |
|
R2 |
-0.005 |
108 |
4.64E+02 |
|
|
R3 |
-0.003 |
105 |
-3.20E+01 |
|
|
Mean |
0.000 |
99 |
2.30E+03 |
99 |
|
SD |
0.008 |
3.62E+03 |
|
||
100
|
R1 |
-0.001 |
102 |
-9.76E+02 |
|
R2 |
0.000 |
100 |
9.44E+02 |
|
|
R3 |
0.000 |
100 |
1.55E+03 |
|
|
Mean |
0.000 |
101 |
5.07E+02 |
100 |
|
SD |
0.001 |
|
1.32E+03 |
|
R1– R6= Replicates 1 to 6
SD= Standard Deviation
[Increase in growth as compared to controls]
Growth curves are shown below.
Description of key information
The 72h-ErL10 and 72h-ErL50 are 12 and 18 mg/L respectively in an OECD guideline 201 test with Pseudokirchnerella subcapitata under GLP (reliability 1).
Key value for chemical safety assessment
- EC50 for freshwater algae:
- 18 mg/L
- EC10 or NOEC for freshwater algae:
- 12 mg/L
Additional information
The ecotoxicity of Eucalyptus citriodora towards Pseudokirchnerella subcapitata was investigated according to OECD guideline 201 under GLP. Algal cells were exposed to WAFs with nominal loading rates of 6.25, 12.5, 25, 50 and 100 mg/L and were observed for 72 hours. The 72h-ErL10 and 72h-ErL50 were found to be 12 and 18 mg/L respectively based on nominal concentrations.
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