Essential oil of Cistus ladaniferus L (Cistaceae) obtained from stems and leaves by distillation

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Deviations:

yes

Remarks:

minor deviations considered to have no impact on the conclusion of the study for more information see Details on study design

GLP compliance:

yes (incl. QA statement)

Remarks:

23 October 2015

Test material

Test material information

Constituent 1

Reference substance name:

Pin-2(3)-ene

EC Number:

201-291-9

EC Name:

Pin-2(3)-ene

Cas Number:

80-56-8

Molecular formula:

C10H16

IUPAC Name:

2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

Constituent 2

Reference substance name:

2,2,6-trimethylcyclohexan-1-one

EC Number:

219-309-9

EC Name:

2,2,6-trimethylcyclohexan-1-one

Cas Number:

2408-37-9

Molecular formula:

C9H16O

IUPAC Name:

2,2,6-trimethylcyclohexanone

Constituent 3

Reference substance name:

(1α,3α,5α)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptan-3-ol

EC Number:

216-813-0

EC Name:

(1α,3α,5α)-6,6-dimethyl-2-methylenebicyclo[3.1.1]heptan-3-ol

Cas Number:

1674-08-4

Molecular formula:

C10H16O

IUPAC Name:

6,6-dimethyl-2-methylenebicyclo[3.1.1]heptan-3-ol

Constituent 4

Reference substance name:

Endo-1,7,7-trimethylbicyclo[2.2.1]hept-2-yl acetate

EC Number:

200-964-4

EC Name:

Endo-1,7,7-trimethylbicyclo[2.2.1]hept-2-yl acetate

Cas Number:

76-49-3

Molecular formula:

C12H20O2

IUPAC Name:

1,7,7-trimethylbicyclo[2.2.1]hept-2-yl acetate

Constituent 5

Reference substance name:

[1aR-(1aα,7α,7aβ,7bα)]-1a,2,3,5,6,7,7a,7b-octahydro-1,1,4,7-tetramethyl-1H-cycloprop[e]azulene

EC Number:

244-565-3

EC Name:

[1aR-(1aα,7α,7aβ,7bα)]-1a,2,3,5,6,7,7a,7b-octahydro-1,1,4,7-tetramethyl-1H-cycloprop[e]azulene

Cas Number:

21747-46-6

Molecular formula:

C15H24

IUPAC Name:

(1aR,7R,7aS,7bR)-1,1,4,7-tetramethyl-1a,2,3,5,6,7,7a,7b-octahydro-1H-cyclopropa[e]azulene

Constituent 6

Reference substance name:

1aR,4R,4aS,7R,7aS,7bS)-1,1,4,7-tetramethyl-2,3,4a,5,6,7,7a,7b-octahydro-1aH-cyclopropa[e]azulen-4-ol

Cas Number:

577-27-5

Molecular formula:

C15H26O

IUPAC Name:

1aR,4R,4aS,7R,7aS,7bS)-1,1,4,7-tetramethyl-2,3,4a,5,6,7,7a,7b-octahydro-1aH-cyclopropa[e]azulen-4-ol

Constituent 7

Reference substance name:

[1aR-(1aα,4β,4aβ,7α,7aβ,7bα)]-decahydro-1,1,4,7-tetramethyl-1H-cycloprop[e]azulen-4-ol

EC Number:

209-003-3

EC Name:

[1aR-(1aα,4β,4aβ,7α,7aβ,7bα)]-decahydro-1,1,4,7-tetramethyl-1H-cycloprop[e]azulen-4-ol

Cas Number:

552-02-3

Molecular formula:

C15H26O

IUPAC Name:

(1aR,4S,4aS,7R,7aS,7bS)-1,1,4,7-tetramethyl-2,3,4a,5,6,7,7a,7b-octahydro-1aH-cyclopropa[e]azulen-4-ol

Test material form:

liquid

Details on test material:

Name Cistus Organic Oil
Batch no. 1215/2
CAS No. 89997-74-0
EINECS-No. 289-711-7
Purity 100%wt (UVCB substance)
Appearance: orange-yellow mobile liquid
Production date 23. Dec. 2015
Expiry date 22. Dec. 2017
Storage: Room Temperature (20 ± 5°C)

Test animals / tissue source

Species:

human

Strain:

other: reconstructed human Cornea-like Epithelium

Details on test animals or tissues and environmental conditions:

EpiOcular TM tissue Model OCL-212 supplied by MatTek

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

The test item was applied as supplied at the dose of 50 µl

Duration of treatment / exposure:

29 minutes

Duration of post- treatment incubation (in vitro):

A 12 -minute post exposure immersion period at room temperature and 2 hours and 5 minutes post exposure incubation at standard culture conditions.

Number of animals or in vitro replicates:

4 replicates.

Details on study design:

Deviations from the study plan:
- Incubator temperature: The temperature in the incubator reached 38.4°C which is +0.4°C higher than the 37+/-1°C as initially scheduled ==> Considering the results obtained, this deviation is considered to have no impact on the conclusion of the study
- Test system: RhCE tissue used was OCL-212 instead of OCL-200 ==> This reference corresponds to a 12-tissue kit whereas OCL-200 refers to individual tissues or 24-tissue kit. ==> This deviation is considered to have no impact on the conclusion of the study
- Treatment of the results: the true viability % of the tissues treated with the test item was calculated by subtracting the mean viability percents of the test item and the NSC living control instead of their ODs as scheduled in the study plan ==> This deviation is considered to have no impact on the conclusion of the study.
- In the first run, the mean tissue viability of the tissues treated with the positive control substance was slightly > 50% (50.58%), instead of <50% as initially scheduled in the study plan. ==> Considering the results obtained, this deviation is considered as without impact on the conclusion of the study.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

other: mean percent tissue viability

Run / experiment:

first run

Value:

63.14

Negative controls validity:

valid

Positive controls validity:

not valid

Remarks:

percent tissue viability = 50.58%

Results 2

Irritation parameter:

other: mean percent tissue viability

Run / experiment:

Second run

Value:

48.72

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

mean percent tissue viability : 40.30%

In vivo

Irritant / corrosive response data:

Cistus oil is potentially irritating or corrosive to the eye.

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:

other: other experiment to class in category 1 or 2 need to be performed

Conclusions:

Cistus oil is potentially irritating or corrosive to the eye. It has to be identified as potentially requiring classification and labelling according to UN GHS category 2 or category 1.

Executive summary:

In a GLP study according to OECD 492 guideline the irritant and corrosive potential of Cistus oil was evaluated. The method is based on viability of Reconstructed Human Cornea-like Epithelium-(RhCE) when exposed to the test substance. In a first run, the mean tissue viability of the tissues treated with the positive control substance was slightly > 50% (50.58%), instead of < 50%. The positive control was not validated therefore a second run was performed. The mean percent tissue viability of the RhCE replicates treated with the test item was 48.72% versus 40.30% in the positive control (Methyl acetate). In conclusion, under experimental conditions adopted and in accordance with the Regulation EC N° 1272/2008 the test item has to be identified as potentially requiring classification and labelling according to UN GHS category 2 or category 1.