ASSESSMENT OF EYE IRRITATION

Objective To assess the potential of the test material to cause damage to

the conjunctivae, cornea or iris after a single exposure of
indeterminate duration.
Guideline OECD Guidelines for the Testing of Chemicals No. 405
Animal Species: Rabbit
Strain: Albino (outbred strain of New Zealand White)
Body Weight: 2.0 - 3.5kg

Pre - Exposure  Both eyes of each animal were examined on the day
before administration for signs of pre-existing irritation,
reaction or abnormality which would eliminate it from
the study. With the help of light source from
ophthalmoscope.
 The rabbits were at least three months old (12-20wks)
Preparation of test  Liquid test materials are used as supplied unless specific
material instructions concerning dilution or other preparation are
given by the Sponsor.
 The pH of test materials in 1 iquid form is recorded. The
Sponsor is informed of any liquid in the ranges pH 1-2 or
pH 11.5 - 14, which are usually regarded as ocular
irritants without animal testing being necessary.
Preparation pH
Measurement
Undiluted as Supplied
90% v/v aqueous preparation
of the test material

Administration of test  Initially, a single rabbit treated
material  The dose was instilled into the right eye of each animal
by pulling the lower eyelid away from the eyeball to
form a cup into which 0.1 ml of the test material was
dropped.
 The eyelids were gently held together for one second and
then released.
 The left eye remained untreated and used as a
comparison with the treated eye during assessment of
ocular lesions.
 After consideration of the ocular responses in first
animal, 2 additional animals were treated.
 If a corrosive or severe irritant effect is not observed in
the initial test, the irritant or negative response should be
confirmed using up to two additional animals.
 If an irritant effect is observed in the initial test, it is
recommended that the confirmatory test be conducted in
a sequential manner in one animal at a time, rather than
exposing the two additional animals simultaneously.
  If the second animal reveals corrosive or severe irritant
effects, the test is not continued.
 If results from the second animal are sufficient to allow
for a hazard classification determination, then no further
testing should be conducted.
Observation period  Ocular reactions to treatment were assessed one hour, 24,
48 and 72 hours after treatment.
 Rabbits showing unresolved lesions on Day 4 may be
pre-examined on Day 8 and at seven-day intervals for up
to 21 days.
 The condition of all treated eyes is checked daily to
ensure the animal is not subject to infection or undue
distress.
Mortality and  If animals recover from any irritation effects within 72
Termination hours after receiving the substance, the study ends on the
Procedure fourth day.
 If the irritation persists or is considered irreversible, the
animals are kept for further observation until the
irritation improves or reaches a point where it cannot be
reversed.
 Animals experiencing severe pain or intense eye effects
are immediately removed from the study and euthanized
to prevent suffering.
 Animals that die during the study or are euthanized are
examined to determine the cause of their condition.
 Animals that survive the study's observation period are
not subjected to further examination of their tissues
unless the study sponsor specifically requests it before
the study starts.

Apparatus available  Pencil beam torch

to facilitate inspection  Ophthalmoscope
of eyes  Hand slit lamp (Remidio PSL-D20 Handheld portable
slit lamp)
 Rabbit No. & Sex 1 - Male 1 - Female
Time After Treatment 1hr 24hr 48hr 72hr 1hr 24hr 48hr 72hr
CORNEA
E = Degree of Opacity
F = Area of cornea
involved
Score (E*F)*2
IRIS
D
Score D*5
CONJUNCTIVAE
A = Redness
B = Chemosis
C = Discharge
Score (A+B+C)*2
Total Score
 Draize Scale for Scoring Ocular Irritation

1. CONJUNCTIVAE
A) Redness (refers to palpebral and bulbar conjunctivae; excluding cornea and
iris)
Normal 0
Some blood vessels hyperaemic (injected) 1
Diffuse,crimson colour; individual vessels not easily discernible. 2
Diffuse beefy red 3
B) Chemosis
Swelling (refers to lids and/or nictating membranes)
0
Normal
1
Any swelling above normal 2
Obvious swelling, with partial eversion of lids 3
Swelling, with lids about half closed 4
Swelling, with lids more than half closed
C) Discharge
No Discharge 0
Any amount different from normal (does not include small amounts observed in 1
inner canthus of normal animals)
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs a considerable area around the 3
eye.
THE TOTAL SCORE = (A+B+C)*2 MAX TOTAL
= 20
2. IRIS
D) Values
Normal 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal 1
hyperaemia; or injection; iris reactive to light (a sluggish reaction is considered 2
to be an effect)
Hemorrhage, gross destruction, or no reaction to light 3
THE TOTAL SCORE = D*5 MAX TOTAL
= 10
3. CORNEA
E) Opacity: degree of density (readings should be taken from most dense area)*
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); 1
details of iris clearly visible
Easily discernible translucent area; details of iris slightly obscured 2
Opaque cornea; iris not discernible through the opacity 3
F) Area of Cornea Involved
One quarter (or less) but not zero
Greater than one quarter but less than half
Greater than half but less than three quarters
Greater than three quarters, up to whole area