Dodecanoic acid, 3-[[2-(2-hydroxyethoxy)ethyl]imino]-2,2-dimethylpropyl ester

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-03-31 to 2009-07-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD Guidelines for Testing of Chemicals, Section 4, No. 431, “In Vitro Skin Corrosion: Human Skin Model”

Version / remarks:

2004

Deviations:

no

Test guideline 2

Guideline:

other: Commission Regulation (EC) No 440/2008, Annex Part B, B.40Bis: “In Vitro Skin Corrosion: Human Skin Model Test”, Official Journal of the European Union No. L142

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-{[2-(2-hydroxyethoxy)ethyl]imino}-2,2-dimethylpropyl dodecanoate

EC Number:

700-067-2

Cas Number:

931419-77-1

Molecular formula:

C21H41NO4

IUPAC Name:

3-{[2-(2-hydroxyethoxy)ethyl]imino}-2,2-dimethylpropyl dodecanoate

Test material form:

liquid

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

The EPISKIN model has been validated for corrosivity testing in an international trial, it is considered to be suitable for this study

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: The EPISKIN Standard Model™ from Skinethic (France) is a three-dimensional human skin model comprising a reconstructed epidermis with a functional stratum corneum.
- Tissue batch number: 09-EKIN-013

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: Each skin unit was rinsed to remove the test item by use of PBS solution (0.9 % NaCl) and returned to the plate in the culture medium.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/mL
- Incubation time: 3 hours
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA

- Barrier function: IC 50 = 2.1 mg/mL
- Morphology: differentiated epidermis consisting of a basal layser, several spinous and granular layers and a thick stratum corneum
- Contamination: no
- Reproducibility: proven

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE - not used

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA
If the mean value is >35% and the variability is less that 50% = Non Corrosive
If the mean value is <35% and the variability is less that 50% = Corrosive

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 50 µL

NEGATIVE CONTROL
- Amount(s) applied: 50 µL
- Concentration: 9 g/L saline

POSITIVE CONTROL
- Amount(s) applied: 50 µL
- Concentration:

Duration of treatment / exposure:

4 hours at room temperature (18-28°C)

Duration of post-treatment incubation (if applicable):

none

Number of replicates:

3 disks

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1st experiment

Value:

148

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: The test to verify if the test item was capable of inducing the colour reaction produced a visible blue precipitation, which indicates that residual test item within the rinsed skin disk could have resulted in a higher OD value
- Colour interference with MTT: not tested

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In this in vitro EPISKIN model test with Sika Hardener LG, the results indicate that the test item is not skin corrosive.

Executive summary:

Disks of EPISKIN were treated with positive control, negative control or test item and incubated for 4 hours. After rinsing, viability of each disk was assessed by use of an MTT method. Viability below 35% of the negative control is considered to indicate a corrosive effect. The positive and negative control results were found to meet the acceptability criteria. The positive control result showed zero viability and the negative control result was at least greater then the OD of the extraction solution alone. The test item did not show a reduced cell viability in comparison to the negative control. In this in vitro EPISKIN model test with Sika Hardener LG results indicated that the test item is not a skin corrosive.