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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to OECD TG 437 and in accordance with the Principles of Good Laboratory Practices (GLP)

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
Remarks:
Minor exceptions which did not have any adverse impact on the study were noted
Principles of method if other than guideline:
not applicable
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: clear, colorless, semi-viscous liquid
Details on test material:
- Name of test material (as cited in study report): 1,4 Cyclohexanedimethanol Reaction Products with Epichlorohydrin
- Analytical purity: The purity of the test material was determined to be 9.5 area % 1,4-Cyclohexanedimethanol monoglycidiyl ether, 45.0 area %, 1,4-Cyclohexanedimethanol diglycidyl ether, and 25.0 area % of the reaction product of 1,4-cyclohexanedimethanol with 3 equivalents of epichlorohydrin by gas chromatography/flame ionization detector with tentative identification of the major components by nuclear magnetic resonance and gas chromatography mass spectrometry
- Impurities (identity and concentrations): no information available
- Lot number : Lot # 1H1050NZP1
- Expiration date of the lot/batch: March 05, 2015

Test animals / tissue source

Species:
other: bovine
Strain:
not specified
Details on test animals or tissues and environmental conditions:
not applicable

Test system

Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
750 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:
The liquid test substance 1,4-Cyclohexanedimethanol, reaction products with epichlorohydrin was tested neat (i.e., undiluted, as received). An aliquot of 750 μL of the test substance, positive control, or negative control was introduced into the anterior chamber while slightly rotating the holder to ensure uniform distribution over the cornea. Three corneas were incubated in the presence of the positive control at 32 ± 1ºC for 10 minutes. Three corneas were incubated in the presence of the negative control at 32 ± 1ºC for 10 minutes. Five corneas were incubated in the presence of the test substance at 32 ± 1ºC for 10 minutes. After the 10-minute exposure time, the control (both positive and negative) or test substance treatments were removed. The epithelial side of the corneas was washed at least three times with Complete MEM (containing phenol red) to ensure total removal of the control or test substances. As residual test substance was observed on the corneas and it appeared to be oily, one additional rinse with Complete MEM (with phenol red) was performed on all corneas to attempt to remove the residual test substance. The corneas were then given a final rinse with Complete MEM (without phenol red). The anterior chambers were refilled with fresh Complete MEM (without phenol red) and an opacity measurement was performed. The corneas were returned to the incubator for approximately 2 hours after which a final measure of opacity was obtained.

After the final opacity measurement was performed, the medium was removed from both chambers of the holder. The posterior chamber was filled with fresh Complete MEM and 1 mL of a 4 mg/mL fluorescein solution was added to the anterior chamber. The corneas were then incubated in a horizontal position (anterior side up) for approximately 90 minutes at 32 ± 1ºC. At the end of the 90-minute incubation period, the medium was removed from the posterior chamber and placed into tubes numbered to correspond with chamber number. Aliquots of 360 μL from the numbered tubes were placed into their designated wells on a 96-well plate. The optical density at 490 nm (OD490) was determined using a Molecular Devices Vmax kinetic microplate reader. If the OD490 value of a control or test substance sample was 1500 or above, a 1:5 dilution of the sample was prepared in Complete MEM without phenol red (to bring the OD490 value within the linear range of the plate reader). A 360 μL sample of each 1:5 dilution was transferred to a clean specified well on the 96-well plate. The plate was read again.

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Remarks:
mean
Run / experiment:
10 min exposure, undiluted
Value:
-1.2
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation

In vivo

Irritant / corrosive response data:
The in vitro score obtained for the test substance, 1,4-Cyclohexanedimethanol, reaction products with epichlorohydrin, was -1.2. The results of the positive control fell within two standard deviations of the historical mean (within a range of 39.2 to 63.9), the assay was considered valid.
Other effects:
not applicable

Any other information on results incl. tables

BCOP Results

Test substance 

Conc. 

Exposure time 

Mean opacity value 

Mean OD490 value 

In vitro score 

pH 

1,4-Cyclohexanedimethanol, reaction products with epichlorohydrin 

Undiluted 

10 minutes 

-1.3 

0.006 

-1.2 

5.0 

Ethanol 

10 minutes 

42.0 

0.747 

53.2 

- 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The in vitro score obtained for the test substance was -1.2. Based upon the results of the study, the test substance, 1,4-Cyclohexanedimethanol, reaction products with epichlorohydrin, is not classified as an eye corrosive or a severe eye irritant (Globally Harmonized System - GHS Category 1) according to the current OECD test guideline (OECD TG 437).
Executive summary:

The Bovine Corneal Opacity and Permeability Assay (BCOP) was used to assess the potential ocular irritancy/toxicity of the test substance, 1,4-Cyclohexanedimethanol, reaction products with epichlorohydrin (1,4-CHDM DGE). Bovine corneas, obtained as a by-product from freshly slaughtered animals, were mounted in special holders and exposed to the test substance for 10 minutes. An in vitro score was determined for the test substance based on the induction of opacity and permeability (to fluorescein) in the isolated bovine corneas. The in vitro score obtained for the test substance was -1.2. Based upon the results of the study, the test substance, 1,4-Cyclohexanedimethanol, reaction products with epichlorohydrin, is not classified as an eye corrosive or a severe eye irritant (Globally Harmonized System - GHS Category 1) according to the current OECD test guideline (OECD TG 437). Furthermore, the test substance would be classified as a non-irritant to eye according to the classification system established by Vanparys et al., where a substance that induces an in vitro score between 0 and 3 would be predicted to be a non-irritant to eye.