12-oxahexadecan-16-olide

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

12-oxahexadecan-16-olide

EC Number:

229-755-6

EC Name:

12-oxahexadecan-16-olide

Cas Number:

6707-60-4

Molecular formula:

C15H28O3

IUPAC Name:

1,6-dioxacycloheptadecan-7-one

Test material form:

liquid

Specific details on test material used for the study:

name of testing material : Cervolide
Batch:VE00445215
Expiry date: 07 June 2018

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: SkinEthic Laboratories, Lyon, France.

Details on animal used as source of test system:

This model is a three-dimensional human epidermis model, which consists of adult human-derived epidermal keratinocytes which have been seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. The keratinocytes were cultured for 13 days, which results in a highly differentiated and stratified epidermis model comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.

Justification for test system used:

In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimize the need of in vivo testing. One of the validated in vitro skin irritation tests is the EPISKIN test, which is recommended in international guidelines (e.g. OECD and EC).

Vehicle:

unchanged (no vehicle)

Details on test system:

On the day of receipt the tissues were transferred to 12-well plates and pre-incubated with pre-warmed Maintenance Medium for 3.5 hours at 37°C. Maintenance medium and Assay medium were supplied by Skinethic Laboratories, Lyon, France.MTT concentrate (Sigma Aldrich, Zwijndrecht, The Netherlands; 3 mg/ml in PBS) diluted (10x) in Assay medium (final concentration 0.3 mg/ml).

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

undiluted (25 µl) directly

Duration of treatment / exposure:

15 ± 0.5 minutes

Duration of post-treatment incubation (if applicable):

After a 42 hour post-incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

83

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

The positive control had a mean cell viability of 8.8% after 15 ± 0.5 minutes exposure. The absolute mean OD₅₇₀(optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was less than 17%, indicating that the test system functioned properly.

Skin irritation is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with CERVOLIDE compared to the negative control tissues was 83%. Since the mean relative tissue viability for CERVOLIDE was above 50% after 15 ± 0.5 minutes treatment CERVOLIDE is considered to be non-irritant.

In conclusion, CERVOLIDE is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with CERVOLIDE compared to the negative control tissues was 83%. Since the mean relative tissue viability for CERVOLIDE was above 50% after 15 ± 0.5 minutes treatment CERVOLIDE is considered to be non-irritant.

Executive summary:

The objective of this study was to evaluate CERVOLIDE for its ability to induce skin irritation on a human three dimensional epidermal model (EPISKIN Small model (EPISKIN-SM^TM)). The possible skin irritation potential of CERVOLIDE was tested through topical application for 15 minutes.  The study procedures described in this report were based on the most recent OECD and EC guidelines.

The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with CERVOLIDE compared to the negative control tissues was 83%. Since the mean relative tissue viability for CERVOLIDE was above 50% after 15 ± 0.5 minutes treatment CERVOLIDE is considered to be non-irritant.

In conclusion, CERVOLIDE is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report.