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Administrative data

Description of key information

SPE1415 was tested in two in vitro assays for skin effects and gave negative results for both corrosion and irritation. It was also tested in two in vitro assays for eye effects. One of the studies (BCOP) gave an inconclusive finding however the other study (RhCE) gave a positive finding.




Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29/4/15 to 5/5/15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
GLP compliance:
yes (incl. certificate)
Species:
other: EpiSkin Reconstructed Human Epidermis Model Kit
Strain:
not specified
Details on test animals and environmental conditions:
Test System: EPISKIN Reconstructed Human Epidermis Model Kit
Supplier: SkinEthic Laboratories, Lyon, France
Date Received: 8th April 2015
EpiSkin Tissues (0.38 cm^2) lot number: 15-EKIN-017
Maintenance Medium lot number: 15-MAIN3-017
Assay Medium lot number: 15-ESSC-017
Type of coverage:
not specified
Preparation of test site:
not specified
Vehicle:
unchanged (no vehicle)
Controls:
not required
Duration of treatment / exposure:
15 minutes
Observation period:
42 hours
Number of animals:
Not applicable
Irritation / corrosion parameter:
other:
Value:
98.3
Remarks on result:
other:
Remarks:
Basis: other: Prediction. Time point: 42 hours. Reversibility: no data. (migrated information)
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The substance was non-irritant to the skin in an in vitro assay
Executive summary:

Introduction

The purpose of this test was to evaluate the skin irritation potential of the test item using the EPISKINTM reconstructed human epidermis model after a treatment period of 15 minutes followed by a post-exposure incubation period of 42 hours. The principle of the assay was based on the measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to the test item by means of the colorimetric MTT reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt (3-[4,5-dimethylthiazol-2 -yl]-2,5-diphenyl-tetrazolium bromide) to a blue formazan salt (within the mitochondria of viable cells) in the test item treated tissues relative to the negative controls.

Method

Triplicate tissues were treated with discs of the test item for an exposure period of 15 minutes. At the end of the exposure period the test item discs were removed from each tissue and each tissue was rinsed before incubating for 42 hours. At the end of the post-exposure incubation period each tissue was taken for MTT-loading. The maintenance medium from beneath each tissue was transferred to pre-labeled micro tubes and stored in a freezer for possible inflammatory mediator determination. After MTT-loading a total biopsy of each epidermis was made and placed into micro tubes containing acidified isopropanol for extraction of formazan crystals out of the MTT-loaded tissues.

At the end of the formazan extraction period each tube was mixed thoroughly and duplicate 200 µL samples were transferred to the appropriate wells of a pre-labeled 96-well plate. The optical density was measured at 562 nm.

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).

Results

The relative mean viability of the test item treated tissues was 98.3% after the 15-Minute exposure period and 42 hours post-exposure incubation period.

Quality criteria: The quality criteria required for acceptance of results in the test were satisfied.

Conclusion

The test item was classified as non-irritant. The following classification criteria apply:

EU DSD and CLP Not classified for Irritation.

UN GHS Not classified for Irritation (category 3 can not be determined).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18/11/15 to 20/11/15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
other: OECD Guideline for Testing of Items 492: Reconstructed human Cornea-like Epithelium (RhCE) test method for identifying items not requiring classification and labelling for eye irritation or serious eye damage
GLP compliance:
yes (incl. certificate)
Species:
other: EpiOcular cornea epithelial model
Strain:
other: Not applicable
Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
A disc of the substance was applied to the tissue culture surface
Duration of treatment / exposure:
6 hours
Observation period (in vivo):
Not applicable
Number of animals or in vitro replicates:
Not applicable
Irritation parameter:
other: Relative mean viability (%)
Basis:
other: Epiocular tissue
Remarks on result:
other: The treated tissues had a relative mean viability of 4.2% following an exposure period of 6 hours
Irritant / corrosive response data:
The treated tissues had a relative mean viability of 4.2% following an exposure period of 6 hours. A score of less than 60% gives a prediction of irritant.
Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The substance gave a response in an RHE assay which predicts as irritant.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

On the basis of the available studies there is no justification for classification for skin effects. However on the basis of the positive result in one in vitro assay, the substance is classified for eye irritancy effects (Eye Irritant, Category 2).