Commiphora myrrha, ext.

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The experiment was performed between 09 November 2017 and 16 November 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted in compliance with OECD Guideline No. 439 without any deviation.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Version / remarks:

Version 28 July 2015

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

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:

SOURCE OF 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

In vitro test system

Test system:

human skin model

Source species:

other:

Cell type:

other: reconstructed epidermis

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- The 0.50 cm² reconstructed epidermises (Episkin SA, RHE/S/17 Batch No. 17-RHE-116) were received on 13 November 2017. The same day, the insert (filter + epidermis) was gently removed
from the agarose while avoiding leaving agarose on the polycarbonate filter. The inserts were placed in 6 wells culture plate which had been previously filled with 1 mL of growth medium (Episkin SA, batch No. 17 SGM 033) during 2 hours and 05 minutes. Then just before treatment, the inserts were placed in 24 wells culture plate which had been previously filled with 300 μL of maintenance medium (Episkin SA, batch No. 17 SMM 022). The two killed Reconstructed Human epidermis control tissue models, supplied by Episkin (Batch No. 17-RHE-021) were defrost on 21 February 2017, re-freeze on 21 February 2017 and defrost to be used on 28 February 2017.

TREATMENT
- The test item was applied, as supplied, at the dose of 16 µL, to the epidermal surface of 3 living human skin models and 2 killed Reconstructed Human epidermis (for non-specific MTT reduction control) during 42 minutes at room temperature. To ensure a good contact with the epidermises, during all the treatment period, the test item was recovered with a nylon mesh provided by Episkin SA.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: Test item was applied for 42 minutes at room temperature
- Temperature of post-treatment incubation: 41 hours and 50 minutes post-treatment incubation period in fresh medium at 37 °C, 5% CO2

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 16 µL
- Concentration (if solution): Undiluted

Duration of treatment / exposure:

42 minutes at room temperature.

Duration of post-treatment incubation (if applicable):

41 hours and 50 minutes post-incubation period at 37 °C, 5% CO2

Number of replicates:

3 living human skin models and 2 killed Reconstructed Human epidermis (for non-specific MTT reduction control)

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Main test (42 minutes)

Value:

92.5

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:

VIABILITY
- The mean corrected percent viability of the treated tissues was 92.5%), versus 1.2% in the positive control (5% Sodium Dodecyl Sulfate).

Any other information on results incl. tables

Table 7.3.1/1: Assessment of the skin irritation - individual and average values of OD after 42 minutes exposure

 

Items	Skin	OD	Mean OD/disc#	Mean OD/product	Viability%	Mean viability%	SD	Conclusion
Negative control	1	0.958	1.073	1.001	107.2	100.0	8.7	-
		1.129
		1.133
	2	1.009	1.1026		102.5
		1.018
		1.052
	3	0.885	0.904		90.3
		0.912
		0.917
Positive control	4	0.010	0.010	0.012	1.0	1.2	0.2	Irritant
		0.011
		0.011
	5	0.011	0.011		1.1
		0.011
		0.011
	6	0.014	0.014		1.4
		0.015
		0.014
Test item	7	1.012	1.062	0.926	106.1	92.5	16.7	Non irritant
		1.128
		1.046
	8	0.771	0.739		73.8
		0.733
		0.713
	9	1.065	0.976		97.5
		0.765
		1.098

 

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

OD: optical density

Acceptance criteria: SD ≤ 18%

Negative control: OD value of the 3 replicates in the range ≥ 0.8 and ≤ 3.0.

The optical density was measured after a 1:2 dilution of the formazan extracts in isopropanol; the acceptability criteria should be in the range ≥ 0.4 and ≤1.5 for the negative control.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under these experimental conditions and in accordance with the Regulation (CE) No.1272/2008 and in absence of information on a skin corrosion, the test item has not to be classified.

Executive summary:

The aim was to evaluate the possible irritating effects of the test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle after topical application on in vitro human reconstructed epidermis (SkinEthic RHE® model).

The test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle was applied as supplied at the dose of 16 μL, to 3 living Reconstructed Human epidermis (SkinEthic RHE® model) during 42 minutes. The application was followed by a rinse with 25 mL of DPBS and a 41 hours and 50 minutes post-incubation period at 37°C, 5% CO2. Cell viability was then measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that was quantitatively measured after extraction from tissues. The experimental protocol was established in accordance with O.E.C.D. Test Guideline No. 439 adopted 28 July 2015 and the Test method B.46 of Council regulation No. 761/2009 dated 23 July 2009 (EU Journal L220) - ATP Council regulation No. 440/2008 of 30

May 2008 (E.U. Journal L142).

The mean percent viability of the treated tissues was 92.5% versus 1.2% in the positive control (5% Sodium Dodecyl Sulfate).

In accordance with the Regulation EC No. 1272/2008, the test item HUILE ESSENTIELLE MYRRHE SOMALIE 100% pure et naturelle has to be considered as Non-irritant to skin. It corresponds to UN GHS No Category.

No hazard statement or signal word is required.