butyl 4-hydroxycyclohexanecarboxylate

Skin irritation / corrosion

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

other: SOP SkinEthic Skin Irritation Test-42bis

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

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 RHE^TMwere 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