Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013-03-18 to 2013-07-25
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: SOP SkinEthic Skin Irritation Test-42bis
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
butyl 4-hydroxycyclohexanecarboxylate
EC Number:
941-637-2
Cas Number:
1384257-92-4
Molecular formula:
C11H20O3
IUPAC Name:
butyl 4-hydroxycyclohexanecarboxylate
Test material form:
other: liquid

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Justification for test system used:
The human skin RHE model has been validated for this type of study.
Vehicle:
unchanged (no vehicle)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 16 µL

NEGATIVE CONTROL
- PBS buffer
- Amount(s) applied (volume or weight): 16 µL

POSITIVE CONTROL
- Sodium dodecyl sulfate
- Amount(s) applied (volume or weight): 16 µL
- Concentration (if solution): 5%
Duration of treatment / exposure:
42 minutes
Duration of post-treatment incubation (if applicable):
not applicable
Number of replicates:
3 membranes for each group (test item, negative control and positive control)

Test animals

Species:
other: Reconstructed human epidermis model
Details on test animals or test system and environmental conditions:
Reconstructed human epidermis model (RHE)
Supplier: SkinEthic
Test System: Human keratinocytes on a polycarbonate filter inserted in a sealed 24-well plate
Tissue Batch: 13 022A 0308
The tissue batch used fully met the quality and validity criteria as described in OECD TG 439

Test system

Type of coverage:
open
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
16 µL
Duration of treatment / exposure:
42 minutes (according to the suppliers protocol)
Number of animals:
The test item, the positive and the negative control were tested in batch-triplicates, i.e. 9 tissues were used for this study
Details on study design:
The study was performed according to OECD TG 439, Council Regulation (EC) No. 761/2009, Method B.46 and the SOP SkinEthic Skin Irritation Test-42bis (2009)

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Value:
90.33
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
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:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes (mean OD = 2.183 at 570 nm)
- Acceptance criteria met for positive control: Yes (mean relative viability = 1.00%)
- Acceptance criteria met for variability between replicate measurements: yes (standard deviation values < 18%)

In vivo

Irritant / corrosive response data:
not applicable
Other effects:
not applicable

Any other information on results incl. tables

This in vitro study was performed to assess the irritating potential of the test item by means of the Human Skin Model Test. The test item was applied topically to a human reconstructed skin model followed by a cell viability test. Cell viability was measured by dehydrogenase conversion of MTT into a blue formazan salt, which was quantitatively measured after extraction from tissues. The percent reduction of cell viability in comparison to untreated negative controls was used to predict skin irritation. Triplicates of the human skin model RHETMwere treated with 16 µL of the test item, the negative (PBS buffer) or the positive control (5% aqueous dodecyl sulphate solution) for 42 minutes, respectively.

Treatment with the positive control induced a sufficient decrease in the relative absorbance as compared to the negative control for the treatment interval thus ensuring the validity of the test system. After treatment with the negative control the absorbance values reached the required acceptability criterion of an optical density (OD) >/=1.2 and </=2.5 for the treatment interval thus showing the quality of the tissues.

The mean relative tissue viability after treatment with the test item was 90.33% and, thus, higher than 50.0%. Therefore, the test item is considered to possess no irritating potential to the skin.

Under the experimental conditions reported, the test item is not irritating to the skin.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
Under the experimental conditions reported, the test item is not irritating to the skin.
Executive summary:

see above