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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:
18 December 1996 to 12 June 1997
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
the certificate of analysis is missing from the study report

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
24 February 1987
Deviations:
yes
Remarks:
temperature and relative humidity occasionally reaching 16; 23°C and 38% respectively. This deviation neither affected the course of the study nor its results.
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
29 December 1992
Deviations:
yes
Remarks:
temperature and relative humidity occasionally reaching 16; 23°C and 38% respectively. This deviation neither affected the course of the study nor its results.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: solid granular powder
Details on test material:
- Other: beige brown-coloured granular powder
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: B&K Universal G.J., S.L. (Sant Vincenç dels Horts, Barcelona, Spain)
- Age at study initiation: approximately 10-11 weeks
- Weight at study initiation: 2.2-2.5 kg
- Housing: Animals were housed individually in stainless steel cages (48 x 59 x 37 cm) with a grille floor, placed on racks.
- Diet: free access to a standard rabbit diet UAR 112 (Panlab, S.L., Barcelona, Spain. Expiry date: 1997-05-29) which had been analyzed by the manufacturer
- Water: ad libitum by an automatic drinking system. The water was supplied by "Compania de Aguas de Sabadell, S.A." and is periodically checked for the presence of possible contaminant.
- Acclimation period: at least seven days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-22, occasionally reaching values of 16 and 23°C
- Humidity (%): 40-70, occasionally reaching levels of 38%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12 (7:00 to 19:00)

IN-LIFE DATES: From: 07 January 1997 To: 20 January 1997

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: the untreated left eye of each animal was used as a control
Amount / concentration applied:
0.1 g (0.1 mL)
Observation period (in vivo):
1 hour, 24, 48 and 72 hours after treatment. An additional reading was performed 5 days after administration. At the end of this period, the observations were ended.
Number of animals or in vitro replicates:
3 male rabbits
Details on study design:
PRIOR TO TREATMENT: both eyes of each animal were examined in order to reject any animal showing eye alterations.

ADMINISTRATION OF TEST SUBSTANCE: the test substance was instilled undiluted in a single application. Animals were immobilized and the test material was poured into the right conjunctival sac made perfectly accessible by carefully pulling out the lower lid. After the application, the lids of the treated eye were held closed for approximately one second.
Initially, a single animal was treated. As no severe ocular alterations were induced, the remaining two animals were also administered the test substance.
Immediately following administration, the behaviour of the animals was observed before being returned to their cages.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no washing

SCORING SYSTEM: outlined in the OECD guideline n° 405

Cornea
Opacity: degree of density (readings should be taken from most dense area)
No ulceration or opacity .......................................................................................................................0
Scattered or diffuse areas of opacity (apart from slight loss of normal lustre);
details of iris clearly visible...................................................................................................................1
Easily discernible translucent area; details of iris slightly obscured.....................................................2
Nacrous area; details of iris not visible; size of pupil barely discernible ..............................................3
Opaque cornea; iris not discernible through the opacity ..................................................................... 4

Iris
Normal ...................................................................................................................................................... 0
Deeply marked folds, congestion, swelling, moderate circumcorneal hyperaemia; or injection,
any of these or combination of any thereof iris still reacting to light (slow reaction is positive)...............1
No reaction to light, hemorrhage, serious damage (any or all of these)....................................................2

Conjunctivae
Redness (refers to palpebral and bulbar conjunctivae; cornea and iris)
Blood vessels normal .................................................................................................................. 0
Some blood vessels hyperaemic (injected) ................................................................................. 1
Diffuse, crimson red; individual vessels difficult to discern ........................................................ 2
Diffuse beefy red ........................................................................................................................ 3

Edema: lids and/or nictating membranes
No swelling ................................................................................................................................... 0
Any swelling above normal (include nictating membranes).......................................................... 1
Obvious swelling, with partial eversion of lids ............................................................................. 2
Swelling, with lids about half closed ............................................................................................. 3
Swelling, with lids nearly or completely closed............................................................................. 4

TOOL USED TO ASSESS SCORE:
Direct examination was performed in the following order: conjunctivae (edema, redness, discharge); iris and cornea. Observation of the cornea was then completed in the readings at 24 and 48 hours after treatment by applying a 2% aqueous sodium fluorescein solution (FLUORESCEINA, Laboratorios Cusi, S.A., batch no.J2) to test the area and then washing the area with a 0.9% physiological saline solution. Once surplus fluorescein had been removed, the corneal alterations were observed with the aid of a transilluminator with a cobalt blue filter.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 48 h
Remarks on result:
other: a score of 0 was obtained at the 1h observation time point
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 48 h
Remarks on result:
other: a score of 0 was obtained at the 1h observation time point
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Remarks on result:
other: a score of 0 was obtained at the 1h observation time point
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Remarks on result:
other: a score of 0 was obtained at the 1h observation time point
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Remarks on result:
other: a score of 0 was obtained at the 1h observation time point
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.67
Max. score:
1
Reversibility:
fully reversible within: 72 h
Remarks on result:
other: a score of 2 was obtained at the 1h observation time point
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 48 h
Remarks on result:
other: a score of 2 was obtained at the 1h observation time point
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 5 days
Remarks on result:
other: a score of 2 was obtained at the 1h observation time point
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.33
Max. score:
1
Reversibility:
fully reversible within: 48 h
Remarks on result:
other: a score of 2 was obtained at the 1h observation time point
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Remarks on result:
other: a score of 3 was obtained at the 1h observation time point that fully reversed at 24h.
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Remarks on result:
other: a score of 1 was obtained at the 1h observation time point that fully reversed at 24h
Irritant / corrosive response data:
- At the 1h observation time point, the test substance induced in the conjunctivae of all the animals diffuse, crimson colour, individual-coloured vessels not easilly discernible (grade 2), accompagnied by lacrimation ranging from mild to wetness of the eyelids and adjacent for, and edematous lesions ranging from mild edeam of the eyelids and nictating membrane (grade 1) to swelling with lids approximately half closed (grade 3).
- At the 24h observation time point, all the animals showed grade 1 for conjuctivae (some clearly hyperaemic blood vessels). Two animals showed grade 1 for opacity (diffuse area affecting at least a quarter of the corneal area), accompanied, in one animal, by slight swelling of the eyelids and nictatingmembrane (grade 1).
- At the 48h observation time point, the lesions had remitted in two animals of the three.
- At the 72h observation time point, only one animal continued to present some clearly hyperaemic blood vessels (grade 1) in the conjuctivae.
- At the 5 days observation time point, the remaining persisting lesion mentioned for the 72h observation time point had completely disappeared.
Other effects:
None. The behaviour and physical condition of the rabbits were normal throughout the study.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the experimental conditions of this study, the test material is not classified as irritating to the eye according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) criteria.
Executive summary:

This GLP-compliant study was performed to assess the potential of the test material to induce eye irritation in vivo, according to OECD Guideline 405 (dated February 24th, 1987) and EU method B.5 of the E.E.C. directive n° 92/69 (dated December 29th, 1992).

Material and methods

0.1 g of the test material were instilled as supplied, into one eye of each of the 3 male rabbits, the other eye untreated serving as control. Ocular examinations were performed on both right and left eyes 1 hour and then 24, 48 and 72 hours after instillation. An additional reading was performed 5 days after administration. At the end of this period, the observations were ended.

Results

Ocular reactions were observed during the study but were all fully reversed 5 days after the administration of the test material. The individual mean scores (24, 48 and 72 hours) were as follows for cornea opacity: 0.33, 0.33 and 0.00, for iris: 0.00, 0.00 and 0.00, for conjunctivae: 0.67, 0.33 and 1.00 and for chemosis: 0.33, 0.00 and 0.00.

Conclusion

Under the experimental conditions of this study, the test material is not classified as irritating to the eye according to the Regulation (EC) No 1272/2008 on classification, labelling and packaging (CLP) and the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) criteria.