Registration Dossier

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
not specified
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
not specified
Qualifier:
according to
Guideline:
other: MatTek Protocol for: INVITRO EpiDermTM SKIN IRRITATION TEST For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm, Model EPI-200-SIT, Rev. 26/3/2012,1-37
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Details on test animals and environmental conditions:
The reconstructed human epidermal model EpiDerm™ (EPI-200, MatTek, Ashland, USA) consists of normal human-derived epidermal keratinocytes, which have been cultured to form a multilayered highly differentiated model of the human epidermis. The EpiDerm™ System is manufactured according to defined quality assurance procedures.
The EpiDerm™ tissues (surface 0.63 cm²) are cultured on specially prepared cell culture inserts and shipped as kits, containing tissues on shipping agarose together with the necessary amount of culture media

Test system

Type of coverage:
other: topical
Preparation of test site:
not specified
Vehicle:
not specified
Details on study design:
The test consists of a topical exposure of the neat test chemical to a reconstructed human epidermis (RhE) model followed by a cell viability test. Cell viability is measured by dehydrogenase conversion of MTT [(3-4,5-dimethyl thiazole 2-yl) 2,5-diphenyltetrazolium-bromide], present in cell mitochondria, into a blue formazan salt that is quantitatively measured after extraction from tissues. The reduction of the viability of tissues exposed to chemicals in comparison to negative controls (treated with PBS) is used to predict the skin irritation potential. Classification of skin irritation potential is performed according to criteria. The viability of tissues is measured as optical density (OD) of the formazan extracts using a spectrophotometer at 570 nm. Relative cell viability is calculated for each tissue as % of the mean OD value of the negative control tissues.

Results and discussion

In vivo

Irritant / corrosive response data:
Under the described experimental design (see any other information on results part), average viability of tissues treated by the test substance was 102.7 % of negative control average value, i.e. viability was > 50 %. The effect of the test substance was negative in EpiDermTM model.
According to the classification criteria, the test substance is considered to have no category in regard to skin irritation.

Any other information on results incl. tables

In vitro alternatives that have been validated and accepted may also be used to help in classification decisions making (see Regulation (EC) 1272/2008, 3.2. Skin corrosion/ irritation, 3.2.2. Classification criteria for substances).

For further classification the relative cell viability is calculated for each tissue as % of the mean of the negative control tissues viability, which is set at 100 %.

The cut-off values for the prediction of irritation are given below; these values are stated in EU Method B.46., part 2.2. INTERPRETATION OF RESULTS:

The test substance is considered to be irritant to skin in accordance with UN GHS 1 category 2:

(i) if the tissue viability after exposure and post-treatment incubation is less than or equal (≤) to 50 %.

The test substance is considered to have no category:

(ii) if the tissue viability after exposure and post-treatment incubation is more than (>) 50 %.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
the test substance is considered to have no category in regard to skin irritation.
Executive summary:

Test substance, 2-methoxy-5-nitrophenyl acetate, was assayed for the in vitro skin irritation in human epidermal model EpiDermTM. The test was performed according to the Method B.46. In vitro skin irritation: Reconstructed human epidermis model test and Protocol for: In Vitro EpiDermTM Skin Irritation Test For use with MatTek Corporation’s Reconstructed Human Epidermal Model EPI-200-SIT (see par. 1.4, (1), (3)). After pre-incubation of tissues, 25 mg of the test was placed directly atop to the previously moistened tissue so it covered all tissue surfaces. Length of exposition was 60 minutes. Three tissues were used for every concentration and controls. After removal of the test substance from tissues, tissues were post incubated for 42 hours due to leave of damage reparation. Three hours incubation with MTT and two hours extraction period with shaking followed then. Optical density (OD570) of isopropyl alcohol extracts was measured on a spectrophotometer. Relative cell viability was calculated for each tissue as % of the mean viability of the negative control tissues. Under the above-described experimental design, average viability of tissues treated by the test substance was 102.7 %, i.e.viability was > 50 %. The effect of the test substance was negative in EpiDermTM model (the tissue was not damaged). According to the classification criteria given in chapter 3.9., the test substance is considered to have no category in regard to skin irritation.