Commiphora myrrha, ext.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The experiment was performed between 19 December 2017 and 21 December 2017,

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study performed according to OECD Guideline 492 without any deviation.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

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)

Version / remarks:

Adopted 28 July 2015

Deviations:

yes

Remarks:

Difference of viability between the 2 tissues treated with the positive control was 31.46%, instead of 20%. Considering the fact that these values are conformed to our historical data, this deviation is without impact on the conclusion of the study.

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Remarks:

27 April 2017

Test material

Test material information

Constituent 1

Reference substance name:

Commiphora myrrha, ext.

EC Number:

284-510-0

EC Name:

Commiphora myrrha, ext.

Cas Number:

84929-26-0

Molecular formula:

Not relevant, UVCB substance

IUPAC Name:

Commiphora myrrha, ext.

Test material form:

liquid

Details on test material:

Batch No.: 0021835
Date received: 17 October 2017
Storage: room temperature, darkness
Form: liquid
Colour: amber-yellow to green-yellow
Production date: June 2017
Best before date: October 2019

Specific details on test material used for the study:

SURCE OF THE TEST MATERIAL
Batch No.: 0021835
Date received: 17 October 2017
Storage: room temperature, darkness
Form: liquid
Colour: amber-yellow to green-yellow
Production date: June 2017
Best before date: October 2019

Test animals / tissue source

Species:

other: reconstructed human cornea-like epithelium tissues

Details on test animals or tissues and environmental conditions:

The 0.60 cm² Reconstructed human Cornea-like Epithelia (EpiOcularTM OCL-212-ver2.0, supplied by MatTek Corporation, batch No. 27018) were received on 19 December 2017.
The same day, the tissues in their well shipping container were equilibrated at room temperature for 15 minutes. Then the inserts (filter + epithelium) were gently removed from the agarose while
avoiding leaving agarose on the polycarbonate filter. They were placed in 6 wells culture plate which had been previously filled with 1 mL of 37°C pre-warmed assay medium (MatTek Corporation, batch No. 121817ISA) and incubated during 19 hours and 55 minutes at standard culture conditions.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 μL
- Concentration (if solution): Undiluted

Duration of treatment / exposure:

30 minutes at 37 °C, 5% CO2, 95% humidity (standard culture conditions).

Duration of post- treatment incubation (in vitro):

After 30 minutes of exposure, extensive rinsing with DPBS at room temperature, a 12 minutes post-exposure immersion period at room temperature and a 2 hours and 03 minutes post-exposure incubation at standard culture conditions.

Number of animals or in vitro replicates:

Test item, negative and positive controls were applied on duplicate tissues (2 living RhCE tissue replicates)

Details on study design:

The 0.60 cm² Reconstructed human Cornea-like Epithelia (EpiOcularTM OCL-212-ver2.0, supplied by MatTek Corporation, batch No. 27018) were received on 19 December 2017.
The same day, the tissues in their well shipping container were equilibrated at room temperature for 15 minutes. Then the inserts (filter + epithelium) were gently removed from the agarose while
avoiding leaving agarose on the polycarbonate filter. They were placed in 6 wells culture plate which had been previously filled with 1 mL of 37°C pre-warmed assay medium (MatTek Corporation, batch No. 121817ISA) and incubated during 19 hours and 55 minutes at standard culture conditions.

Results and discussion

In vitro

Results

Irritation parameter:

other: %mean viability

Run / experiment:

Main test - 30 minutes

Value:

97.37

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

MEAN PERCENT TISSUE VIABILITIES
- The mean corrected percent tissue viability of the RhCE replicates treated with the test item was 97.37 %, versus 18.36% in the positive control (Methyl acetate).
ACCEPTANCE OF RESULTS:
- The mean percent tissue viabilities obtained with the positive control and negative controls are within the range of historical data and therefore validate the experiment.

The difference of viability between the 2 tissues treated with the positive control was 31.46%, instead of 20% at the maximum as initially scheduled.
Considering the results obtained and the fact that these values are conformed to our historical data, this deviation is considered as without impact on the conclusion of the
study.

Any other information on results incl. tables

Table 7.3.2/1: Assessment of the eye irritation potential individual and average values of OD after 30 minutes exposure

 

Items	Tissues	OD	Mean OD/disc#	Mean OD / product	Viability%	Mean viability%	Difference of viability%	Conclusion
Negative control	1	0.920	0.950	0.951	99.95	100.00	0.11	-
		0.952
		0.978
	2	0.933	0.951		100.05
		0.954
		0.967
Positive control	3	0.023	0.025	0.175	2.63	18.36	31.46	UN GHS category 2 or 1
		0.026
		0.027
	4	0.318	0.324		34.09
		0.321
		0.334
Test item	5	0.907	0.945	0.926	99.42	97.37	4.10	-
		0.974
		0.955
	6	0.897	0.906		95.32
		0.916
		0.905

#: mean of 3 values (triplicate of the same extract)

OD: optical density

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the experimental conditions adopted and in accordance with the Regulation EC No. 1272/2008, the test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle does not require classification for eye irritation or serious eye damage. It corresponds to the UN GHS No Category.
No hazard statement and the signal word are required.

Executive summary:

The aim of the study was to evaluate the eye hazard potential of the test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle after topical administration on in vitro reconstructed human cornea-like epithelium tissues (EpiOcularTM tissue model).

The test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle was applied as supplied, at the dose of 50 μL, to 2 living DPBS pre-treated RhCE (EpiOcularTM tissue model) during 30 minutes at 37°C, 5% CO2, 95% humidity (standard culture conditions). The exposure period was followed by extensive rinsing with DPBS at room temperature, a 12 minutes post-exposure immersion period at room temperature and a 2 hours post-exposure incubation at standard culture conditions. The tissue viability was measured by performing an MTT assay. The experimental protocol was established in accordance with O.E.C.D. Test Guideline No. 492 adopted 28 July 2015.

The mean percent tissue viability of the RhCE replicates treated with the test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle was 97.37%, versus 18.36% in the positive control (Methyl acetate). In conclusion, under the experimental conditions adopted and in accordance with the Regulation EC No. 1272/2008, the test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle does not require classification for eye irritation or serious eye damage. It corresponds to the UN GHS No Category.

No hazard statement and the signal word are required.