Registration Dossier

Diss Factsheets

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
Lime Oxide
IUPAC Name:
Lime Oxide
Constituent 2
Reference substance name:
1,6-Octadien-3-ol, 3,7-dimethyl-, acid-isomerized
EC Number:
277-225-8
EC Name:
1,6-Octadien-3-ol, 3,7-dimethyl-, acid-isomerized
Cas Number:
73018-51-6
IUPAC Name:
1,6-Octadien-3-ol, 3,7-dimethyl-, acid-isomerized
Constituent 3
Reference substance name:
Reaction mass of (E)-5-(but-2-en-2-yl)-2,2-dimethyltetrahydrofuran and 1-isopropyl-4-methylcyclohexa-1,3-diene and 1-isopropyl-4-methylcyclohexa-1,4-diene and 1-methyl-4-(propan-2-ylidene)cyclohex-1-ene
IUPAC Name:
Reaction mass of (E)-5-(but-2-en-2-yl)-2,2-dimethyltetrahydrofuran and 1-isopropyl-4-methylcyclohexa-1,3-diene and 1-isopropyl-4-methylcyclohexa-1,4-diene and 1-methyl-4-(propan-2-ylidene)cyclohex-1-ene
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Lime Oxide
- Molecular formula: predominantly a mixture of monoterpenes C10H16 and cyclic monoterpene ethers C10H18O
- Molecular weight: monoterpenes C10H16 molecular weight 136.2, cyclic monoterpene ethers C10H18O molecular weight 154.2
- Physical state: clear colorless liquid
- Analytical purity: 61.8%, sum of 4 major peaks
- Impurities: Known impurities are similar in nature to the major components and have empirical formulae of either C10H16 or C10H18O
- Lot/batch No.: SC00009577
- Expiration date of the lot/batch: 13 October 2015
- Storage condition of test material: room temperature, in the dark
Specific details on test material used for the study:
The test article, a colourless to pale yellow liquid, was identified as Lime Oxide.
A certificate of analysis for the test article was provided.
The test article was administered without dilution.

Test animals / tissue source

Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
Corneas were obtained from bovine eyes supplied by a local abattoir. The eyes were
removed after slaughter, completely immersed in Hanks’ Balanced Salt Solution
(containing penicillin at 100 IU/mL and streptomycin at 100 μg/mL) in a suitably
sized container and transported on the same day to the testing facility.
On arrival at the test facility the eyes were carefully examined for defects including
increased opacity, scratches and neovascularisation. Only corneas free from such
defects were used.
Fresh corneas, mounted onto specifically designed holders were treated topically with
the test material. Eye corrosion / severe irritation potential was based on the combined
effect of the test article on the opacity of the cornea following the treatment and the
cornea’s ability to resist penetration of a fluorescent dye through the tissue.

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
750 μL
Duration of treatment / exposure:
10 minute incubation at 32°C ± 1°C.
Observation period (in vivo):
NA
Duration of post- treatment incubation (in vitro):
NA
Number of animals or in vitro replicates:
three corneas
Details on study design:
A volume of 750 μL of the test article was applied to each of three corneas followed
by a ten minute incubation at 32°C. After this incubation, each cornea was washed
with media containing phenol red (as a pH indicator) until this indicator showed no
pH effect occurring (and demonstrating that the test article had been removed
successfully). The corneas were then washed once in media without phenol red. The
corneas were incubated (horizontally) for 2 hours ± 10 minutes after which corneal
opacity was measured.
For the permeability endpoint, the media in the anterior chamber was replaced with
1 mL of sodium fluorescein (4 mg/mL solution), while the posterior chamber was
filled with fresh MEM. The corneas were incubated in the vertical position for
90 minutes. Following this period, the media in the posterior chamber was removed
and held in a labelled tube. Three 350 μL aliquots of this media (per cornea) were
analysed for optical density at 490 nanometers (OD490) using a spectrophotometer.
A volume of 750 μL of the negative or positive control was similarly applied to
further groups of three corneas. These groups were subject to the procedures detailed
above.

Results and discussion

In vitro

Results
Irritation parameter:
cornea opacity score
Run / experiment:
1
Value:
4.7
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
not determinable
Other effects / acceptance of results:
Corneal Opacity:
The mean corrected opacity reading for the test article was 4.7.
The mean corrected opacity reading for the negative control was 0.0.
The mean corrected opacity reading for the positive control was 39.3.
The corneas treated with the positive control were noted to be cloudy following treatment.

Corneal Permeability:
The mean group corrected optical density for the test article was 0.185.
The mean group corrected optical density for the negative control was 0.0.
The mean group corrected optical density for the positive control was 0.203.

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
The test article, Lime Oxide, produced an IVIS score of 7.45. No prediction can be made in respect of its potential to cause eye irritation.
The assay was considered valid as the assay acceptance criteria were met.
Executive summary:

The test article, Lime Oxide, produced an IVIS score of 7.45.

Categories Display