Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
2020-06-26
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
2019-07-31
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: New guidance document on an integrated approach on testing and assessment (IATA) for skin corrosion and irritation, Series on Testing and Assessment No. 203
Version / remarks:
2014-07-11
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Landesamt für Umwelt, Rheinland Pfalz, Germany
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
The reconstructed human epidermis (RhE) model closely mimics the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis and has been validated, considered scientifically valid and adopted as OECD test guideline. The test is part of a Turnkey Testing Strategy to assess the skin corrosion and irritation potential of the test material including transport classification.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Three-dimensional human epidermis model (EpiDerm 200, MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia)
- Tissue Lot number: 30892

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: At room temperature for 25 minutes, then in the incubator at 37°C for 35 minutes.
- Temperature of post-treatment incubation: 37°C for 24±2 hours, then 37°C for another 18 ± 2 hours

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: The tissues were washed with sterile PBS to remove residual test material after start of application.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: Sunrise Absorbance Reader
- Wavelength: 570 nm
- Filter: filter wavelength 570 nm without reference filter

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: See section "ACCEPTANCE CRITERIA" in "Any other information on materials and methods incl. tables".
- Barrier function: See section "ACCEPTANCE CRITERIA" in "Any other information on materials and methods incl. tables".

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- freeze killed tissues: No
The test substance was added to 0.9 mL MTT solution. The mixture was incubated in the dark at about 37°C for 3 hours. A negative control (deionized water) was tested concurrently. If the color of the MTT solution or (in case of water-insoluble test substances) the border to the water-phase turned blue / purple, the test substance was presumed to reduce MTT directly. In case of direct MTT reduction, three freeze-killed control tissues (KC) were treated additionally with each the test article and the negative control in the same way as described for the test item. Based on the result of the pretest, it was judged that application of killed control tissues is not necessary.

PREDICTION MODEL / DECISION CRITERIA
See section "EVALUATION OF RESULTS" in "Any other information on materials and methods incl. tables".
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 30 µL of undiluted liquid test substance

NEGATIVE CONTROL
- Amount applied: 30 µL

POSITIVE CONTROL
- Amount applied: 30 µL
- Concentration: 5% SDS
Duration of treatment / exposure:
25 minutes at room temperature and 35 minutes at 37°C
Duration of post-treatment incubation (if applicable):
42 hours
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean of 3 tissues
Value:
4.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
tissue 1
Value:
5.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
tissue 2
Value:
4.4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
tissue 3
Value:
4.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Direct MTT reduction: No direct MTT reduction occured.
- Colour interference with MTT: No colour interference occured.

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.

The relative mean viability of the tissues treated with the test substance determined after an exposure period of 1 hour with an about 42-hour post-incubation was 4.7%. The variability between the results of the tissues is within the acceptance range.

The tissues treated with the positive control 5% SDS showed a relative mean viability of 2.5%, reflecting the expected sensitivity of the tissues.

The mean OD570 of the negative control (PBS) fulfills the acceptance criteria and demonstrates the validity of the assay.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2021
Report date:
2021

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
2019-06-18
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: New guidance document on an integrated approach on testing and assessment (IATA) for skin corrosion and irritation, Series on Testing and Assessment No. 203
Version / remarks:
2014-07-11
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Landesamt für Umwelt, Rheinland Pfalz, Germany

Test material

1
Reference substance name:
Santalol oil: fermentation products of glucose with santalene synthase modified Rhodobacter sphaeroides, distilled, oxidised
Cas Number:
2576531-09-2
IUPAC Name:
Santalol oil: fermentation products of glucose with santalene synthase modified Rhodobacter sphaeroides, distilled, oxidised

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
The reconstructed human epidermis (RhE) model closely mimics the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis and has been validated, considered scientifically valid and adopted as OECD test guideline. The test is part of a Turnkey Testing Strategy to assess the skin corrosion and irritation potential of the test material including transport classification.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Three-dimensional human epidermis model (EpiDerm 200, MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia)
- Tissue batch number: 30892

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: for 3 minutes at room temperature or for 1 hour in the incubator at 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: The tissues were washed with sterile PBS to remove residual test material

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: Sunrise Absorbance Reader
- Wavelength: 570 nm
- Filter: filter wavelength 570 nm without reference filter

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: See section "ACCEPTANCE CRITERIA" in "Any other information on materials and methods incl. tables".
- Barrier function: See section "ACCEPTANCE CRITERIA" in "Any other information on materials and methods incl. tables".

NUMBER OF REPLICATE TISSUES: 2

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- freeze- killed tissues: No
The test substance was added to 0.9 mL MTT solution. The mixture was incubated in the dark at about 37°C for 3 hours. A negative control (deionized water) was tested concurrently. If the color of the MTT solution or (in case of water-insoluble test substances) the border to the water-phase turned blue / purple, the test substance was presumed to reduce MTT directly. In case of direct MTT reduction, three freeze-killed control tissues (KC) were treated additionally with each the test article and the negative control in the same way as described for the test item. Based on the result of the pretest, it was judged that application of killed control tissues is not necessary.

PREDICTION MODEL / DECISION CRITERIA
See section "EVALUATION OF RESULTS" in "Any other information on materials and methods incl. tables".
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 50 µL of undiluted liquid test substance

NEGATIVE CONTROL
- Amount applied: 50 µL

POSITIVE CONTROL
- Amount applied: 50 µL
- Concentration: 8 N potassium hydroxide solution
Duration of treatment / exposure:
3 minutes at room temperature or 1 hour in the incubator at 37°C
Number of replicates:
2

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean of 2 tissues, exposure period of 3 min
Value:
107.9
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
tissue 1, exposure period of 3 min
Value:
107.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
tissue 2, exposure period of 3 min
Value:
108.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: no indication of skin corrosion
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean of 2 tissues, exposure period of 1 hour
Value:
122.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: no indication of skin corrosion
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
tissue 1, exposure period of 1 hour
Value:
123.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: no indication of skin corrosion
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
tissue 2, exposure period of 1 hour
Value:
121.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: no indication of skin corrosion
Other effects / acceptance of results:
OTHER EFFECTS:
- Direct MTT reduction: No direct MTT reduction occured.
- Colour interference with MTT: No colour interference occured.

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

The relative mean viability of the tissues treated with the test substance determined after an exposure period of 3 minutes was 107.9%, and it was 122.2% after an exposure period of 1 hour. The variability between the results of the tissues is within the acceptance range.

The tissues treated with the positive control 8 N KOH showed a relative mean viability of the tissues of 7.5% (3-minute exposure) and 5.5% (1-hour exposure), reflecting the expected sensitivity of the tissues.

The mean OD570 of the negative control (deionized water) fulfills the acceptance criteria and demonstrates the validity of the assay.

Applicant's summary and conclusion