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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2014-10-14 to 2014-10-20
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
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
(2013)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayrisches Landesamt für Gesundheit und Lebensmittelsicherheit, München, Germany

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Methyl [3-(trimethoxysilyl)propyl]carbamate
- Physical state: liquid
- Stability under test conditions: stable
- Storage condition of test material: at RT, protect from light, avoid humidity

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: human-derived epidermal keratinocytes (EpiDerm)
Justification for test system used:
This test uses the EpiDerm(TM) reconstructed human epidermis model (MatTek) which consists of human keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely mimics the biochemical and physiological porperties of the upper part of the human, i.e. the epidermis.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm(TM) tissues were provided as kits (EPI-200-SIT, MatTek)

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1°C
- Temperature of post-treatment incubation: 37 ± 1°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: washed intensively with DPBS for 1 min
- Observable damage in the tissue due to washing: no
- Modifications to validated SOP: none

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1mg/mL
- Incubation time: 3 h
- Spectrophotometer: plate spectrophometer
- Wavelength: 550 nm


NUMBER OF REPLICATE TISSUES: 3


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

DECISION CRITERIA
- The test substance is considered to be irritant to skin if the viability after 15 minutes exposure and 42 hours post-exposure is less than or equal to 50%.
- The test substance is considered to be not irritant to skin if the viability after 15 minutes exposure 42 hours post-exposure is greater than 50% and the viability.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 30 µL

NEGATIVE CONTROL
- Amount applied: 30 µL DBPS

POSITIVE CONTROL
- Amount applied: 30 µL
- Concentration: 5% (v/v)
Duration of treatment / exposure:
60 min
Duration of post-treatment incubation (if applicable):
42 h
Number of replicates:
3

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
test group
Value:
96.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: no
- Direct-MTT reduction: no
- Colour interference with MTT: no

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

Any other information on results incl. tables

EXPERIMENTAL RESULTS:

 

Negative control

Positive control

Test item

Tissue sample

1

2

3

1

2

3

1

2

3

OD550

1.592

1.564

1.451

1.499

1.449

1.391

0.044

0.044

0.038

0.037

0.037

0.038

1.540

1.563

1.429

1.395

1.343

1.340

OD550(mean)

1.578

1.475

1.420

0.044

0.038

0.038

1.552

1.412

1.342

SD

0.080

0.003

0.107

OD550(mean values of replicates)

1.491

0.040

1.435

Viability (%)

100

2.7

96.2

Applicant's summary and conclusion

Interpretation of results:
other: CLP/EU GHS criteria are not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
In an in vitro skin irritation human skin model test (EpiDerm) according to OECD guideline 439 and in compliance with GLP, a cell viability of 96.2% was measured after 60 min exposure and 42 h post-exposure time when compared to the untreated control. Therefore, the test item should be classified as non-irritant to the skin.