LUMIPOD

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08/11/2011 - 22/11/2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

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

Constituent 1

Reference substance name:

L-Leucine, reaction products with 1,4:3,6-dianhydro-D-glucitol, iso-Pr alc. and octanoyl chloride

Molecular formula:

Not applicable

IUPAC Name:

L-Leucine, reaction products with 1,4:3,6-dianhydro-D-glucitol, iso-Pr alc. and octanoyl chloride

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from a single donor

Source strain:

other: PK2KTU06

Vehicle:

not specified

Details on test system:

Test system:
The present study has been performed using SkinEthic Reconstituted Human Epidermal model (RHE) from Skin Ethic laboratories (Lyon, France), which is validated for skin irritation testing.
The statement regarding validity of in vitro tests for skin irritation testing from the ECVAM Scientific Advisory Committee (ESAC) is joined in annex 1.
Epidermal tissues were received on November 15, 2011, by Elodie Bombard, at day 18 of the differentiation process (air-lifted cultures).
Epidermal tissue viability and responsiveness were ensured by the supplier, as well as expiry dates.
The technical and quality data sheet from SkinEthic Laboratories presented in annex 2 confirmed the good shape and viability of the epidermis. Before the beginning of the experiments, the Study Director checked all documents and the integrity of tissues and culture media (maintenance medium and growth medium).

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

16 + 0.5 µl

Duration of treatment / exposure:

42min (+-1min) at room temperature.

Duration of post-treatment incubation (if applicable):

RHE were then incubated for 42h (+-1h) at 37°C, 5% CO, for recovery.

Number of replicates:

3 replicates by test item and reference

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Mean of three runs with two replicates each

Value:

ca. 1

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Results

1. MTT interference

These data were presented in the specific study reports.

 

2. Acceptance criteria and test validity

Negative control (NC) acceptance criteria: The NC data meet the acceptance criteria if the mean OD value of the 3 tissues is >= 1.2 at 570 nm. The standard deviation value is considered as valid if it is =< 18%.

Positive control (PC) acceptance criteria: The PC data meet the acceptance criteria if the mean viability, expressed as %of the NC, is < 40 % and the standard deviation value is =< 18%.

Batch acceptance criteria: All test items data from one batch are considered as valid if both negative and positive controls data fulfill the above criteria requirements.

Acceptance criteria	Standard deviation value	Validity of the test
NC acceptance criteria	o.7%	Yes
PC acceptance criteria	0.2 %	Yes
Batch acceptance criteria	/	Yes

3. MU viability testing

A MTT assay was performed to measure the viability of RHE after a 42 min exposure to the test and reference items and a 42h recovery period.

The mean OD of the three tissues exposed to the negative control (PBS) was set to represent icio % of tissue viability and the results were expressed as a percentage of the negative control viability.

According to the 4^th revised edition of the Globally Harmonized System of Classification and Labelling of Chemicals (61-15; United Nation (New York and Geneva, 2011)), the prediction model is as follows:

Criteria for in vitro interpretation	Classification GHS United Nation
Mean tissues viability is =< 50 %	Category 2, irritant (I)
Mean tissues viability is > 50 %	No Category, non irritant (NI)

 

Applicant's summary and conclusion

Conclusions:

The mean viability of the tissues treated with the test item is 1%. According to the prediction model described in the 4" revised edition of the GHS, the test item is irritant.