[No public or meaningful name is available]

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07 December 2017 to 15 December 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

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:

Cetyl Acetate and Stearyl Acetate and Oleyl Acetate and Acetylated Lanolin Alcohol

Molecular formula:

C18H36O2-C20H38O2-C20H40O2-C29H48O2

IUPAC Name:

Cetyl Acetate and Stearyl Acetate and Oleyl Acetate and Acetylated Lanolin Alcohol

Test material form:

other: paste

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

other: reconstructed human skin

Cell source:

other: EpiDerm TM (EPI-200-MatTek Corporation)

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm TM (EPI-200-MatTek Corporation)
- Tissue batch number(s): (Lot 25864)
- Delivery date: 12 December 2017
- Date of initiation of testing: no data

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2)
- Temperature of post-treatment incubation: 42 hour post-dose incubation at 37°C, 5% CO2

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: not indicated (according to MatTEek SOP)

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE: no data (according to MatTEek SOP)
- MTT concentration: not specified
- Incubation time: no data
- Spectrophotometer: not identified measured at 570 nm
- Wavelength: 570 nm
- Filter: none
- Linear OD range of spectrophotometer: no data

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: yes (1.601 ± 0.007 (mean and SD of MTT value of 3 tissues exposed to H2O) )
- Barrier function: yes (ET 50% of tissues exposed to 100 µL Triton X-100 1% (n=3): 5.84 h)
- Morphology: yes ( Histological examination demonstrates human epidermis-like structure: including multiple layers (at least 4) of viable epithelial cells (basal layer, stratum spinosum, stratum granulosum) which are present under multilayered stratum corneum ; Tissue thickness: 128 µm )
- Contamination: No contamination reported
- Reproducibility: yes (The mean OD of the 6 wells containing extraction solvent alone (blanks) should be ≤ 0.1., but was 0.149 evaluated not to have affected the measurements, based upon historical system data for blanks; < 0.244)

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µl

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl PBS

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µl SDS
- Concentration (if solution): 5% in water

Duration of treatment / exposure:

60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2)

Duration of post-treatment incubation (if applicable):

42 hour post-dose incubation at 37°C, 5% CO2

Number of replicates:

3/treatment

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Value:

106.9

Negative controls validity:

valid

Positive controls validity:

valid

Remarks:

mean viability 4.5%

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

No colour interference or interference with MTT reduction as tested in a pre-test

Any other information on results incl. tables

Name	Code	Mean of OD	SD of OD	Mean of viability (%)	SD of viability (%)	CV %	Classification
DPBS	NC	1.768	0.210	100.000	11.904	11.904	Non-Irritant
SDS 5%	PC	0.080	0.016	4.535	0.909	20.046	Irritant
Substance	TA2	1.889	0.142	106.873	8.008	7.493	Non-Irritant

 

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The substance is non-irritant in the EpiDermTM model

Executive summary:

The substance was tested for skin irritation in the EpiDermTM model according to OECD 439. 30 uL of the substance, positive control (5% SDS) and negative control (PBS) was applied to the tissue for 60 minutes (25 minutes at room temperature and 35 minutes at 37°C, 5% CO2) and thereafter incubated for 42 hour post-dose at 37°C, 5% CO2. The substance did not interfere by colour or by direct MTT reduction. Tissue viability was 106.9% of negative controls, while validity criteria were evaluated to be met. The substance is concluded to be non-irritant.