Silybum marianum, 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:

06 September 2016 - 09 September 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 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Silybum marianum, ext.

EC Number:

283-298-7

EC Name:

Silybum marianum, ext.

Cas Number:

84604-20-6

Molecular formula:

Not available - UVCB substance

IUPAC Name:

Extract of Chardon marie without support

Test material form:

liquid

Details on test material:

Name: Extract of Chardon marie without support
Batch/Lot number: VDO06205A
Appearance: Light yellow brown cloudy oil
Purity: Considered as 100%
Retest date: 13 July 2019
Storage conditions: Refrigerated
Safety precautions: Routine safety precautions (lab coat, gloves, safety glasses, face mask) for unknown materials were applied to assure personnel health and safety.


Name: Extract of Chardon marie without support
Batch/Lot number: ABT01059A
Appearance: Brown/orange turbid oil
Purity: Considered as 100%
Retest date: 19 July 2018
Storage conditions: Refrigerated
Safety precautions: Routine safety precautions (lab coat, gloves, safety glasses, face mask) for unknown materials were applied to assure personnel health and safety.

Reference: MPB-693061 / MPF-00024688
Batch number: VDO06126E
Provider: Pierre Fabre
Reference: GD 1508
Certificate of analysis: Yes
MSDS: Yes
Type: Raw material
Form: Liquid
Nature: Complex mixture
Colour: Yellowish
Storage conditions: Stored at room temperature
Requested test conditions: To be tested at 100 % (as is)
Sent quantity: 10 g
Expiry / Retest date: 27 November 2016

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Foreskin

Source strain:

not specified

Details on animal used as source of test system:

Information not available

Justification for test system used:

Recommended in the OECD test guidelines

Vehicle:

unchanged (no vehicle)

Details on test system:

The evaluation of the in vitro cutaneous tolerance was performed on reconstructed human epidermis SkinEthicTM RHE model (0.5 cm2) provided by SkinEthic Laboratories.
Every unit of the reconstructed epidermis consists of a stratified and differentiated epidermis obtained from human keratinocytes. Cells are grown on inert polycarbonate filter on chemically defined medium, for 17 days.
Reconstructed epidermis was maintained in agar medium supplied by SkinEthic Laboratories during the transport. On receipt of the epidermis kits on day 17, the colour of the agar medium was checked. The epidermises were transferred onto 2 ml of growth medium, and incubated at least 2 hours (37°C, 5% CO2)

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Test material: 16 μl
Negative control: 16 μl of PBS
Positive control: 16 μl of SDS at 5 % in water

Duration of treatment / exposure:

42 min ± 1 min at room temperature

Duration of post-treatment incubation (if applicable):

Recovery period at 37±3 °C, 5±1% CO2: 42 hours ± 1 hour

Number of replicates:

3 replicates of the test item, negative controll and positive control.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean

Value:

103.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Remarks:

standard deviation 6.2%

Other effects / acceptance of results:

Solubility test
The requested test conditions specify that the test item has to be tested at 100%, thus, no solubility assay was performed.

Condition of test item for exposure
The requested test conditions specify that the test item has to be tested at 100%, thus, no dilution was made. The test item was tested neat and, at this step, weighing data of the test item were documented.
The application of test item was achieved with a positive displacement pipette and the solutions were spread uniformly over the surface of the epidermises by the application of a nylon mesh.

MTT-interaction of the test item
No direct interaction of the test substance with the solution of MTT was observed.

Colouring potential of the test item
The test item did not color the tissues nor perturb the OD measurements during the MTT assay.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The viability mean of the tissues treated with the test item MPB-693061 / MPF-00024688 at 100 % was measured to be higher than 50% by in vitro evaluation of acute skin irritation using the reconstructed human epidermis SkinEthicTM RHE model, therefore the test item MPB-693061 / MPF-00024688 is classified as potentially non-irritant.

Executive summary:

The skin irritation potential was tested in vitro using the reconstructed human epidermis model SkinEthicTM RHE which in its overall design closely mimics the biochemical and physiological properties of the upper parts of the human skin.

 

For that purpose, cell viability of the epidermis was measured by enzymatic conversion of the vital MTT dye into a blue formazan salt that is quantitatively measured after extraction from tissues after exposure of the epidermises to the test item for a standardized

time period.

 

Relative viability was evaluated by comparison to a negative control and expressed as a percentage (%). Irritant test substances were identified by their ability to decrease cell viability below defined threshold levels.

 

This method was granted regulatory approval for skin irritation testing: OECD (2015),Test No. 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method, OECD Guidelines for the Testing of Chemicals, Section 4, OECD Publishing, Paris.

The viability mean of the tissues treated with the test item MPB-693061 / MPF-00024688 at 100 % was measured to be higher than 50% byin vitroevaluation of acute skin irritation using the reconstructed human epidermis SkinEthicTM RHE model, therefore the test item MPB-693061 / MPF-00024688 is classified as potentially non-irritant.