Oils, cinnamon

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18/09/2014 - 29/05/2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study according to international guideline (OECD guideline 202) under GLP. No deviations from guideline.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Principles of method if other than guideline:

Not relevant

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

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

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: water accomodated fractions (WAF) with nominal loading rates of 0.291, 0.64, 1.41, 3.10, 6.82 and 15.0 mg/l
- Sampling method: Analytical samples were taken from each test item concentration and control at 0 hours (initial value) and after 48 hours from aged test solutions in the main test.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the low water solubility of the test item, test item solutions (≥ 1.00 mg/L) were prepared by directly weighing of each “loading rate” (concentration) (15.0, 6.82, 3.10, 1.41 mg/L) into test medium. 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 at least one hour or until the phases have 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. To prepare test solutions with test item concentrations of 0.640 and 0.291 mg/L, adequate aliquots of the 15 mg/L withdrawn phase were diluted with test medium.
- 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):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus, Clone V
- Source: Umweltbundesamt in Berlin/Germany
- Age at study initiation (mean and range, SD): Less than 24 hours
- Method of breeding: Daphnia magna was reared as single culture where one daphnid is kept per 100 mL Elendt M4 medium. The pH-value of the aerated water was within a range of 6.0 – 9.0. The dissolved oxygen was above 60 % saturation and the total hardness 140 - 250 mg/L (as CaCO3). The animals were fed with single cell green algae (Desmodesmus subspicatus, formerly Scenedesmus subspicatus) at least three times a week. The daphnids were reared at a temperature of 20 ± 2 °C in a climatic chamber with 16 hours of illumination and 8 hours of darkness. The medium was changed three times per week. A pipette was used to separate the young daphnids from the adults.
- Feeding during test: No

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

Not relevant

Test conditions

Hardness:

14°dH corresponding to 250 mg/L CaCO3

Test temperature:

20.3 – 21.7 °C

pH:

7.28 – 7.76

Dissolved oxygen:

8.0 mg/L ± 0.4 mg/L

Salinity:

Not relevant

Nominal and measured concentrations:

Nominal: 0.291, 0.64, 1.41, 3.10, 6.82 and 15.0 mg/l (WAF)
Measured (t=0h, fresh): < LoQ, Measured (t=48h, aged):

Details on test conditions:

TEST SYSTEM
- Test vessel: Bottle
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: Glass, 100 ml, filled with 100 ml test medium and minimum headspace
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Elendt M4 medium
- Culture medium different from test medium: No
- Intervals of water quality measurement: The test temperature and the pH-value as well as the oxygen concentration of the test media were measured at all concentrations at t = 0 and 24 h from fresh test solutions and at 24 and 48 h from aged test solutions in one additional replicate per loading rate and control in the non-GLP range finding test and the main test.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: No exposure to light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): immobility (at 24 and 28 hours)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Range finding study
- Test concentrations: 1.0, 10.0 and 100 mg/l (WAF)
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

14.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL = 13.0 - 15.6 mg/l

Details on results:

- Mortality of control: 0%

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: 100%
- EC50/LC50: 1 mg/l < 24h-EC50 < 2.0 mg/l

Reported statistics and error estimates:

The 24 h and 48 h EL50 are the estimated loading rate where 50 % of the daphnids were immobilised after 24 and 48 hours, respectively. As the immobility at the highest loading rate was only at 25 % after 24 hours, no statistical evaluation was indicated. For the evaluation of the 48 h EL50, probit analysis following Gompertz procedure was used. The evaluation of data was performed by using SAS® Proprietary Software Version 9.3. The NOEL was established based on the highest loading rate at which the immobilisation is not higher than the allowed control immobilisation (=< 10 % immobilisation).

Any other information on results incl. tables

Immobilisation after 24 hours

Test item loading rate (mg/l)	Control	0.291	0.640	1.41	3.10	6.82	15.0
	Immobilised daphnids after 24h
Group 1	0	0	0	0	0	0	2
Group 2	0	0	0	0	0	0	1
Group 3	0	0	0	0	0	0	1
Group 4	0	0	0	0	0	0	1
Sum	0	0	0	0	0	0	5
Percentage (%)	0	0	0	0	0	0	25

Immobilisation after 48 hours

Test item loading rate (mg/l)	Control	0.291	0.640	1.41	3.10	6.82	15.0
	Immobilised daphnids after 48h
Group 1	0	0	0	0	0	1	3
Group 2	0	0	0	0	0	0	3
Group 3	0	0	0	0	0	1	3
Group 4	0	0	0	0	0	0	2
Sum	0	0	0	0	0	2	11
Percentage (%)	0	0	0	0	0	10	55

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Less than 10% mortality in controls, Oxygen concentration above 3 mg/l

Conclusions:

The acute toxicity (48h-EL50) of Cinnamon bark oil towards Daphnia magna is 14.2 mg/l.

Executive summary:

The acute toxicity of Cinnamon bark oil towards Dahnia magna was investigated according to OECD guideline 202 under GLP. Daphnids were exposed to WAFs with nominal loading rates of 0.291, 0.640, 1.41, 3.10, 6.82 and 15.0 mg/l and were observed for 48 hours. Based on nominal loading rates, the 24h-EL50 and 48h-EL50 were found to be >15.0 and 14.2 mg/l respectively.