4,4-Dimethyl-2-undecyl-2-oxazoline

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-01-19 to 2010-02-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

ECVAM protocol validated in April 2007 (version 1.2 of September 2005, 1.8 of February 2009)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

equivalent or similar 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:

4,4-dimethyl-2-undecyl-4,5-dihydro-1,3-oxazole

EC Number:

686-853-5

Cas Number:

46921-17-9

Molecular formula:

C16H31NO

IUPAC Name:

4,4-dimethyl-2-undecyl-4,5-dihydro-1,3-oxazole

impurity 1

Reference substance name:

Unknown impurities

IUPAC Name:

Unknown impurities

Test material form:

liquid

Details on test material:

Other name of 4,4-Dimethyl-2-undecyl-2-oxazoline : cycloceramide

Specific details on test material used for the study:

Purity : 96,3 %

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Reconstructed human epidermis from SkinEthic Laboratories company

Source strain:

not specified

Details on animal used as source of test system:

Kit EPISKIN (reconstructed human epidermis) constituted:
- Type I collagen matrix surfaced with a type IV collagen film.
- stratified and differentiated epidermis derived from human keratinocytes, obtained after 13 days of culture.

Justification for test system used:

The in vitro test (EPISKIN model) is a reliable and relevant model for predicting skin irritation and can be used as an in vivo test replacement to distinguish between skin irritants and non-irritants according to EU classification standards.

Vehicle:

unchanged (no vehicle)

Details on test system:

- pH of the test material: not applicable because the test material is lipophilic
- Duration of exposure: 15 +/- 0.5 minutes
- Deposit quantity: 10µL
- Post exposure observation period: incubation; 42 +/- 1 hour / Coloration: 3 hours +/- 5 minutes
- Control group: positive control: PBS / negative control: SDS (CAS number 151-21-3) at 5% in distilled water
- vehicle: no
- removal of test substance: 25 mL of PBS
-criteria:
Viability < 50% : irritant
Viability > 50% and IL-1a > 60 pg/mL : irritant
Viability > 50% and IL-1a < 60 pg/mL : non irritant

Control samples:

yes, concurrent no treatment

yes, concurrent positive control

Amount/concentration applied:

MTT: final concentration of the test medium: 0.3 mg/mL.
Deposit of 10µL of test material or control (positive or negative) in 90µL of water.

Duration of treatment / exposure:

15 +/- 0.5 minutes

Duration of post-treatment incubation (if applicable):

Post-treatment incubation (CO2) : 42 +/- 1 hours

Number of replicates:

2 tests per item (positive control, negative control, test material) and 3 replicates per test

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Remarks:

MTT

Run / experiment:

1

Value:

ca. 58.9

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Remarks:

IL-1a concentration = 185,6 pg/mL

Results 2

Irritation / corrosion parameter:

% tissue viability

Remarks:

MTT

Run / experiment:

2

Value:

ca. 56.5

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Remarks:

IL-1a concentration = 173,1 pg/mL

Any other information on results incl. tables

Test material	Test	% relative viability (dosage of MTT)	final concentration of IL-1a
No vehicle	1	58.9 +/- 0.02	185,6 pg/mL
	2	56.5 +/- 0.00	173.1 pg/mL
	Average	57.70 +/- 1.70	179.35 +/- 8.84 pg/mL

Applicant's summary and conclusion

Conclusions:

According to the results obtained under the experimental conditions adopted, the test element called "CYCLOCERAMIDE (R)" applied on reconstituted human epidermis, can be qualified as skin irritant
(4,4-Dimethyl-2-undecyl-2-oxazoline = Cyclocéramide (R))

Executive summary:

objective: to predict and classify the cutaneous irritating potential of an ingredient, by evaluation of its effect on a human epidermis reconstructed model EPISKIN, in agreement with the standards of European Classification.

Conclusion: According to the results obtained under the experimental conditions adopted, the test element called "CYCLOCERAMIDE (R)" applied on reconstituted human epidermis, can be qualified as skin irritant.

Test material	Test	% relative viability (dosage of MTT)	final concentration of IL-1a
No vehicle	1	58.9 +/- 0.02	185,6 pg/mL
	2	56.5 +/- 0.00	173.1 pg/mL
	Average	57.70 +/- 1.70	179.35 +/- 8.84 pg/mL