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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 13 July 1998 (Start of in-life phase) to 30 September 1998 (GLP compliance statement)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
deviations from the maximum level of daily mean relative humidity : see additional data in results' section
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
not specified
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material: Cesium Fluoro Aluminate Complex
- Purity: 99.5%
- Batch: 604/013
- Physical state: White amorphous powder
- Storage condition of test material: At room temperature in the dark
- Expiration date of the lot/batch: 22 May 2001

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: 7 weeks old
- Weight at study initiation: body weight was less than 1.5 kg
At least prior to dosing, it was ensured that the animal was healthy and that the eyes to be treated were intact and free from any abnormality.
- Housing: Individually housed in labeled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
- Diet: Standard laboratory rabbit diet (LKK-20, pellet diameter 4 mm, Hope Farms, Woerden, The Netherlands) approx. 100 grams per day. In addition, hay (BMI, Helmond, The Netherlands) was provided once a week.
- Water : Free access to tap water diluted with decalcified water.
- Acclimation period: at least 5 days before start of treatment under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature: controlled at 21°C
- Humidity: controlled at 50 %
- Air changes : approximately 15 room air changes/hour
- Photoperiod : 12-hour light/12-hour dark cycle
Fluctuations from these optimal conditions were noted, but were considered as not to have affected study integrity.

IN-LIFE DATES: From: 13 July to 3 August 1998.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 97.8 mg (approx. 0.1 ml).

VEHICLE
- Not applicable.
Duration of treatment / exposure:
Single instillation.
Observation period (in vivo):
21 days.
Number of animals or in vitro replicates:
One male.
Details on study design:
APPLICATION OF THE TEST SUBSTANCE:
On day 1, 97.8 mg of the test substance (a volume of approx. 0.1 ml) was instilled in the conjunctival sac of an eye of one animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control.

REMOVAL OF TEST SUBSTANCE
Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.
Immediately after fluorescein observation on day 2, the treated eye was rinsed with approx 50 mL tap water, using a velocity of flow which did not affect the eye, to remove any visible residual test substance. For reference control the other eye was also rinsed.

SCORING SYSTEM:
The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded.The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

- CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal luster) .........................................0
Scattered or diffuse areas of opacity, details of iris clearly visible................................................. 1
Easily discernible translucent area, details of iris slightly obscured...............................................2
Nacreous area, no details of iris visible, size of pupil barely discernible........................................3
Opaque cornea, iris not discernible through the opacity .................................................................4

Area of cornea involved:
No ulceration or opacity...........................................................................................................................0
One quarter or less but not zero ...........................................................................................................1
Greater than one quarter, but less than half ........................................................................................2
Greater than half, but less than three quarters ...................................................................................3
Greater than three quarters, up to whole area ....................................................................................4

- IRIS
Normal .......................................................................................................................................................0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia,
or injection, any of these or combination thereof, iris still reacting to light
(sluggish reaction is positive) ................................................................................................................1
No reaction to light, hemorrhage, gross destruction (any or all of these)......................................2

- CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal ...............................................................................................................................0
Some blood vessels definitely hyperaemic (injected)..........................................................................1
Diffuse, crimson color, individual vessels not easily discernible......................................................2
Diffuse beefy red.......................................................................................................................................3

Chemosis (refers to lids and/or nictitating membranes):
No swelling ………………………………………… ....................................................................................0
Any swelling above normal (includes nictitating membranes) .........................................................1
Obvious swelling with partial eversion of lids......................................................................................2
Swelling with lids about half closed .......................................................................................................3
Swelling with lids more than half closed ...............................................................................................4

- Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals) ........0
Any amount different from normal and/or lacrimation .....................................................................1
Discharge with moistening of the lids and hairs just adjacent to lids ..............................................2
Discharge with moistening of the lids and hairs (considerable area around the eye) ...................3

TOOL USED TO ASSESS SCORE:
Where standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp. In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
2.7
Max. score:
3
Reversibility:
not fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
3.3
Max. score:
4
Reversibility:
not fully reversible within: 21 days
Irritant / corrosive response data:
Irritation
Instillation of 97.8 mg cesium fluoro aluminate complex (a volume of approximately 0.1 mL) into one eye of one rabbit resulted in marked effects on the cornea, iris and conjuctivae. The corneal injury consisted of opacity (maximum grade 2) and epithelial damage (maximum 75% of the corneal area). As a result of the corneal injury, pannus (neovascularization of the cornea) was apparent 7 days after instillation onwards. Iridic irritation grade 1 was observed which had resolved within 14 days. The irritation of the conjunctivae consisted of redness, chemosis and discharge. Reduced elasticity of the eye lids was noted 14 and 21 days after instillation. Signs of necrosis on the eyelids and nictitating membrane were apparent 7 days after instillation and persisted on the nictitating membrane at termination. In addition, corneal injury (opacity grade 1 and 25% epithelial damage), pannus and irritation of the conjunctivae persisted until termination. Remnants of the test substance were present in the eye on day 1 and 2.

Based on the degree and persistence of the corneal leasions, it was concluded that ocular corrosion had occurred by instillation of the test substance into the rabbit eye.
Other effects:
Coloration / Remnants:
Remnants of the test substance were present in the eye on Day 1. No staining of (peri) ocular tissues by the test substance was observed.

Toxicity / Mortality:
No symptoms of systemic toxicity were observed in the animal during the test period and no mortality occurred.

Any other information on results incl. tables

Table 1: Individual eye irritation scores

Cornea

Iris

Conjunctivae

Discharge

Comments

Animal

Time after

dosing

Opacity

(0-4)

Area

(0-4)

Fluor area (%)

(0-2)

Redness

(0-3)

Chemosis

(0-4)

Discol*

(0-3)

1152

1 hour

1

4

 

1

2 (ABC)

4 (AB)

-

2

b

 

24 hours

1

4

75

1

2 (ABC)

4 (AB)

-

3

b

 

48 hours

1

2

-

1

3 (A)

3 (A)

-

2

-

 

72 hours

1

2

50

1

3 (AB)

3 (A)

-

2

-

 

7 days

2

1

50

1

3 (AB)

3 (A)

AB

2

p

 

14 days

2

1

35

0

3 (AB)

1 (ABC)

AB

1

pf

 

21 days

1 

1 

25 

0 

3 (AB) 

1 (A) 

B

1 

pf

Comments:

The test substance could only partly be removed by rinsing the treated eyes immediately after the 24 hours observation period.

b Remnants of the test substance in the eye

* Grey/white discolouration of the tissue indicated, as a sign of necrosis

p Pannus (neovascularisation of the cornea)

f Reduced elasticity of the eyelids

Score given fore:

A=Eyelids

B=Nictitating membrane

C=Sclera

Applicant's summary and conclusion

Interpretation of results:
Category 1 (irreversible effects on the eye)
Conclusions:
Based on the degree and persistence of the corneal leasions, it was concluded that ocular corrosion had occurred by instillation of cesium aluminium fluoride – complex into the rabbit eye.
Executive summary:

The acute eye irritation/corrosion of cesium aluminium fluoride - complex in the rabbit was investigated in a study performed according to the OECD Testing Guideline No.405 and under GLP.

A single sample of 97.8 mg of cesium fluoro aluminate complex (a volume of approximately 0.1ml) was instilled into an eye of one rabbit. Observations were made 1, 24, 48 and 72 hours and 7, 14 and 21 days after instillation. Vocalisation was noted immediately after instillation, indicating pain. Instillation of Cesium fluoroaluminate complex resulted in marked effects on the cornea, iris and conjuctivae.

The corneal injury consisted of opacity (maximum grade 2) and epithelial damage (maximum 75% of the corneal area). As a result of the corneal injury, pannus (neovascularization of the cornea) was apparent 7 days after instillation onwards. Iridic irritation grade 1 was observed which had resolved within 14 days. The irritation of the conjunctivae consisted of redness, chemosis and discharge. Reduced elasticity of the eye lids was noted 14 and 21 days after instillation. Signs of necrosis on the eyelids and nictitating membrane were apparent 7 days after instillation and persisted on the nictitating membrane at termination. In addition, corneal injury (opacity grade 1 and 25% epithelial damage), pannus and irritation of the conjunctivae persisted until termination. Remnants of the test substance were present in the eye on day 1 and 2. Based on the degree and persistence of the corneal lesions, it was concluded that ocular corrosion had occurred by instillation of cesium aluminium fluoride - complex into the rabbit eye. Based on the results of this study, cesium aluminium fluoride - complex should be classified as category 1 with respect to eye damage/eye irritation.