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Eye irritation

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eye irritation, other
This is in vivo study but correct assignment results in validation failure.
Type of information:
experimental study
Adequacy of study:
key study
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Similar to a guideline study, but the dose was lower, exposure times shorter and washing procedures different than OECD guidelines.

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
other: Haskell Laboratories protocol, E.I. DuPont DeNemours and Company, Wilmington, DE, USA
Principles of method if other than guideline:
Test material is administered into the conjunctival sac of a New Zealand white rabbit, and after various times, the material is removed by washing with various mixtures. Irritation is observed
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
Details on test material:

4,4'-Methylenedicyclohexanamine > 95% purity. Liquid with freezing/melting point of 13 degrees C.
Isomer content (%):
trans-trans 21.22
cis-cis 14.91
cis-trans 59.67
Impurities:2,4'-Methylenedicyclohexanamine: 4.20%
[4-(p-aminobenzyl)cyclohexylamine] methane (PABC): 0.23%
p-aminodicyclohexylmethane (ADCM): 0.05%
p-PABC: 0.038%

Test animals / tissue source

New Zealand White
Details on test animals or tissues and environmental conditions:
males, each weighing 2-3 kg.

Test system

unchanged (no vehicle)
not required
Amount / concentration applied:
0.01 ml. The standard dose of 0.1% ml resulted in clear eye corrosion.
Observation period (in vivo):
60 days
Number of animals or in vitro replicates:
3 per group
Details on study design:
Test material(0.01ml) was placed in the conjunctival sac and washed with tap water for one minute with tap water after 20 seconds or two minutes. The untreated eye served as the control. Observations of cornea, iris and conjunctiva were made with a hand-slit lamp at 1 and 4 hours after administration, at 1,2 and 3 days, and weekly through 60 days after administration. Fluor-i-strip (R) ophthalmic indicators and a biomicroscope were used at examinations 1 and 4 hours after administration. A veterinary ophthalmologist was consulted for selected case examinations. Three animals were sacrificed on day 70 after treatment for histopathologic examination. Eyes were excised, washed, fixed and hardened according to the method of Saunders and Rubin (1975).

Results and discussion

In vivo

Irritation parameter:
other: qualitative assessment
Time point:
other: 60 days
other: 30% reversible within 60 days.
Remarks on result:
other: Dose of 0.01 ml is 1/10th "standard" dose of 0.1 ml.
Irritant / corrosive response data:
Pupillary constriction at 1-4 h, pupillary dilation through day 7, iridial congestion, with injection at 28 days, clouds of precipitate in anterior chamber, diffuse fibrous opacities of the lens, swelling and endothelial relucency by 7 days, corneal vascularization by 14 days, with subsequent healing of the ulceration by 35 days. The conjunctiva showed extreme redness and swelling within 1 h, with moderate to severe discharge (sometimes bloody and purulent).
Other effects:
Histopathology revealed dense corneal scarring, inflammation and lenticular opacity in selected animals. Vascular scarring of cornea can occur. In severe cases, crystals form and clots, with protein and cells present in the anterior chamber, and the lens capsule can rupture.

Any other information on results incl. tables

Each group of 3 rabbits had one rabbit whose eye was normal or nearly normal at 60 days after treatment. One other rabbit had partial healing (partial pannus), and the final rabbit had severe permanent ocular damage. Several rabbits’ eyes developed corneal opacities, still present at 60 days. Histopathology revealed some damage not observed upon gross examination.

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye)
Migrated information corrosive when 0.1 ml is applied in vivo; partially reversible when 0.01 ml is applied and washed in vivo. Criteria used for interpretation of results: EU
4,4'-Methylenedicyclohexanamine causes severe damage to the eye.