Reaction product of a ((2S)-2, 6-diaminohexanoic acid hydrochloride salt, phosphonic acid and methanal) with a 1,2-disubstituted alkane and sodium hydroxide.

Skin irritation / corrosion

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

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)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

other: Light brown solid

Details on test material:

- Name of test material (as cited in study report): SPE1415
- Physical state: Light brown solid
- Analytical purity: Not supplied
- Lot/batch No.: 9
- Expiration date of the lot/batch: 20th February 2016
- Storage condition of test material: Room temperature in the dark

Test animals

Species:

other: EpiSkin Reconstructed Human Epidermis Model Kit

Strain:

not specified

Details on test animals or test system 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

Test system

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

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: other: Relative mean tissue viability

Value:

98.3

Remarks on result:

other:

Remarks:

Basis: other: Prediction. Time point: 42 hours. Reversibility: no data. (migrated information)

Applicant's summary and conclusion

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).