4-methylpent-1-ene

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2011-08-30 to 2011-09-01

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: SkinEthic reconstructed human corneal epithelial model (10-minute exposure)

Deviations:

no

Principles of method if other than guideline:

The test is based on the hypothesis that irritant chemicals are able to penetrate the corneal epithelial tissue and are sufficiently cytotoxic to cause cell death.

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of Inspection: 2011-07-19 to 2011-07-21; Date of signature 2011-08-31

Test material

Test material information

Constituent 1

Reference substance name:

4-methylpent-1-ene

EC Number:

211-720-1

EC Name:

4-methylpent-1-ene

Cas Number:

691-37-2

Molecular formula:

C6H12

IUPAC Name:

4-methylpent-1-ene

Details on test material:

- Name of test material (as cited in study report): 4-methylpent-1-ene
- Physical state: clear colourless liquid
- Analytical purity: ≥ 97 %:
- Lot/batch No.: H1130311
- Date received: 2011-04-11
- Expiration date of the lot/batch: 2011-10-11
- Storage condition of test material: approximately 4 degrees Centigrade in the dark

Test animals / tissue source

Species:

other: not applicable

Strain:

not specified

Details on test animals or tissues and environmental conditions:

The SkinEthic HCE model consists of transformed human corneal epithelial cells of the cell line HCE (LSU EYE Center, New Orleans, USA) that form a corneal epithelial tissue (mucosa), devoid of stratum corneum, resembling histologically the mucosa of the human eye. The model consists of an airlifted, living, corneal tissue construct, produced in polycarbonate inserts in serum-free and chemically defined medium.

Test system

Vehicle:

not specified

Controls:

other: not applicable

Amount / concentration applied:

30 μL

Duration of treatment / exposure:

10 minutes

Observation period (in vivo):

Not applicable

Number of animals or in vitro replicates:

Not applicable

Details on study design:

METHOD
- The test material was applied directly to the culture surface, at the air interface, so that undiluted and/or end use dilutions could be tested directly.
- Cytotoxicity was determined by the reduction of MTT to formazan by viable cells in the test item treated tissues (quantitative measurement of tissue viability) relative to the negative control.
- One tissue for each treatment group was retained for possible tissue histopathology.

ASSESSMENT OF DIRECT TEST ITEM REDUCTION OF MTT
- The MTT assay is a colourmetric method of determining cell viability based on reduction of the yellow tetrazolium salt (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a blue formazan dye by mitochondrial succinate dehydrogenase in viable cells.
- A known limitation of the assay is that test materials may interfere with the MTT endpoint if they are directly reduce MTT and, at the same time, are present on or in the tissues when the MTT viability test is performed. To identify any possible interference, the test item was checked for the ability to reduce MTT.

Results and discussion

In vivo

Results

Irritation parameter:

other: relative viability of treated tissues

Basis:

mean

Time point:

other: 10 minutes

Score:

86.4

Remarks on result:

other: Percent

Irritant / corrosive response data:

No data

Any other information on results incl. tables

DIRECT MTT REDUCTION

- The MTT solution containing the test item turned yellow, which indicated the test material did not directly reduce MTT.

ASSESSMENT OF EYE IRRITATION POTENTIAL

- Individual and mean OD540 values and mean viabilities for each treatment group are given in Table 1 (attached).

 

HISTOPATHOLOGY

- It was considered unnecessary to proceed with tissue histopathology.

 

VALIDITY CRITERIA

- The quality criterion required for acceptance of results was satisfied.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: expert judgment

Conclusions:

The test material was considered to be non-irritating.