Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018
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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
adopted 28. July 2015
Deviations:
not specified
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
adopted 06. Jul. 2012
Deviations:
not specified
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material: 1,3,2-dioxathiane 2,2-dioxide
- IUPAC name: 1,3,2-Dioxathiane 2,2-dioxide
- Molecular formula: C3H6O4S
- Molecular weight: 138.14 g/mol
- Substance type: Organic
- Physical state: Solid

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
other: human-derived epidermal keratinocytes
Cell source:
other: EpiDermTM tissues were procured from MatTek In Vitro Life Science Laboratories, Brati-slava.
Details on animal used as source of test system:
The test system is a commercially available EpiDermTM-Kit, procured by MatTek.
The EpiDermTM tissues are cultured on specially prepared cell culture inserts.
Justification for test system used:
The test consists of a topical exposure of the neat test item to a human reconstructed epi-dermis model followed by a cell viability test. Cell viability is measured by dehydrogenase conversion of MTT 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide, present in cell mitochondria, into a blue formazan salt that is quantitatively measured after extraction from tissues. The percentage reduction of cell viability in comparison of untreated negative controls is used to predict the skin irritation potential.
Vehicle:
unchanged (no vehicle)
Details on test system:
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1 °C
- Temperature of post-treatment incubation (if applicable): 37 ± 1 °C
REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: 1 (rinsing thoroughly with DPBS)
- Observable damage in the tissue due to washing: Blotted with sterile cellulose tissue
DYE BINDING METHOD
- Dye used in the dye-binding assay: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (=MTT)
- Spectrophotometer: Plate spectrophotometer
- Wavelength: 570 nm
NUMBER OF INDEPENDENT TESTING RUNS / EXPERIMENTS TO DERIVE FINAL PREDICTION: 3 (Pre - Tests, Pre-Incubation of Tissues and Treatment)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The tissues were wetted with 25 µL DPBS buffer and spreading it to match the tissue size. The following amounts were applied to the tissues:
1 replicate: 26.4 mg
2 replicate: 26.7 mg
3 replicate: 26.7 mg

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL
- Concentration: DPBS buffer

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL
- Concentration: 5% SDS solution
Duration of treatment / exposure:
44 hours and 10 minutes at 37 ± 1°C and 5.0 ± 0.5% CO2
Duration of post-treatment incubation (if applicable):
43 hours and 35 minutes at 37 ± 1°C and 5.0 ± 0.5% CO2

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
ca. 65.3
Vehicle controls validity:
not valid
Negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test item 1,3,2-Dioxathiane, 2,2-dioxide (EC:214-022-5) is considered non-irritant to skin in the Reconstructed human Epidermis (RhE) Test Method.
Executive summary:

One valid experiment was performed.

Three tissues of the human skin model EpiDermTMwere treated withthe test itemfor 60 minutes.

The test item was applied directly to each tissue and spread to match the tissue size (0.63  cm2; as indicated by the supplier).

DPBS-buffer was used as negative control and 5% SDS solution was used as positive control.

After treatment with the negative control, the mean absorbance value was within the required acceptability criterion of 0.8 <= mean OD <= 2.8, OD was 1.6.

The positive control showed clear irritating effects. The mean value of relative tissue viability was reduced to 3.7% (required:<=20%).

The variation within the tissue replicates of negative control, positive control and test item was acceptable (required: <= 18%).

 

After the treatment with the test item, the mean value of relative tissue viability was reduced to 65.3%. This value is above the threshold for skin irritation potential (50%). Test items that induce values above the threshold of 50% are considered non-irritant to skin.