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Toxicological information

Eye irritation

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Administrative data

Endpoint:
eye irritation, other
Remarks:
points 8.1 and 8.2 of Annex VIII of REACH have been amended. Nevertheless, adequate information from existing in vivo skin irritation or eye irritation studies can still be used to fulfil the information requirement at any tonnage level.
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From the 11th of January to the 8th of February, 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose:
read-across source
Reference
Endpoint:
eye irritation, other
Remarks:
points 8.1 and 8.2 of Annex VIII of REACH have been amended. Nevertheless, adequate information from existing in vivo skin irritation or eye irritation studies can still be used to fulfil the information requirement at any tonnage level.
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From the 11th of January to the 8th of February, 1999
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Justification for Read Across is detailed in the endpoint summary and it is further detailed in the report attached to the IUCLID section 13.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
As required by the Dutch Act on Animal Experimentation, the study protocol was reviewed and agreed by the Article 14-functionary and the Ethical Committee of NOTOX.
Species: Albino Rabbit, New Zealand White, (SPF-Quality)
Source: Charles River Nederland, the Netherlands
Age and body weight: at least 6 weeks old and body weights less than 3.5 kg
Identification: earmark
ENVIRONMENTAL CONDITIONS
Conditions: air-conditioned room with approximately 15 air changes per hour
Temperature: 21°C
Relative humidity of 50%.
Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity.
Lighting: 12 hours artificial fluorescent light and 12 hours dark per day.
Accomodation: Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
Acclimatisation period: at least 5 days before start of treatment under laboratory conditions.
Diet: Standard laboratory rabbit diet (LKK-20, pellet diameter 4mm, Hope Farms, Woerden, The Netherlands) approx. 100 gram per day. In addition, hay (BMI, Helmond, The Netherlands) was provided once a week.
Certificates of analysis were examined and then retained in the NOTOX archives.
Free access to tap-water diluted with decalcified water.
Certificates of quarterly analysis were examined and then retained in the NOTOX archives.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
instillation of 99.6 ± 0.3 mg of the test substance (a volume of approximately 0.1 ml)
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
1, 24, 48 and 72 hours and 7 and 14 days after instillation
Number of animals or in vitro replicates:
3 animals of one sex
Details on study design:
Since marked effects were considered possible this eye irritation study was started by treatment of a single rabbit (sentinel).
The two other animals were treated in a similar manner 14 days later, after considering the degree of eye irritation observed in the first animal except that the eyes of the third rabbit were anaesthesised with a locai anaesteticum (0.4 % Benoxinate Hydroxide, Chravlls Pharmaceuticals, England), approximately 8 minutes prior to instillation.
A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.
Each animal was treated by instillation in the conjunctival sac of one of the eyes 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.
Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. When considered necessary, this procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.
OBSERVATION
Mortality/Viability: twice daily
Toxicity: once a day
Body weights: day of the treatment
Irritation: The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 and 14 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.
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
0.1
Max. score:
1
Reversibility:
fully reversible
Remarks on result:
other: brown coloration
Irritation parameter:
iris score
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
0.1
Max. score:
2
Reversibility:
not fully reversible within: 48 hours
Remarks on result:
other: brown coloration
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
2.9
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks on result:
other: brown coloration
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
1.2
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks on result:
other: brown coloration
Irritant / corrosive response data:
Vocalisation of two rabbits was noted immediately after instillation, indicating pain.
Therefore, the eyes of the third rabbit were anaesthesised with a local anaesteticum (0.4 % Benoxinate Hydroxide, Chravils Pharmaceuticals, England), approximately 8 minutes prior to instillation.
Irritation
Instillation of approximately 99.6 mg of the test substance (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. Corneal injury was seen as opacity (maximum grade 1) and epithelial damage (maximum 25 % of the corneal area) in one animal, which had completely resolved within 72 hours.
Iridial irritation (grade 1) was observed in all animals after 1 hour which recurred in one animal after 48 hours.
Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 14 days in all animals.
Due to brown staining by the test substance, scoring for conjunctival redness was prohibited in all animals after 1 hour and was partly prohibited in one animalup to 72 hours.
Corrosion
There was no evidence of ocular corrosion.
Other effects:
Colouration
Brown staining of the conjunctivae and of the fur on the head and paws, caused by the test substance, was noted during the observation period. Remnants of the test substance were present in the eyes of all animals on day 1 and on the outside of the eyelids of two animals between 1 and 72 hours.
Toxicity/Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.
Interpretation of results:
irritating
Remarks:
Classification criteria according to the CLP Regulation 1272/2008 and its amendments
Conclusions:
Eye irritation was caused by the instillation of the substance.
In addition, brown staining of the conjunctivae and of the fur on the head and paws was noted during the observation period.
Executive summary:

Acute eye irritation/corrosion study on a similar substance 01 was carried out based on the guidelines OECD No.405 "Acute Eye Irritation/Corrosionand the EU B.5, "Acute Toxicity - Eye irritation".

Single samples of approximately 99.6 mg of the similar substance (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours and 7 and 14 days after instillation. 

Vocalisation of two rabbits was noted immediately after instillation, indicating pain. Therefore, the eyes of the third rabbit were anaesthesised with a local anaestheticum. Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae. Corneal injury was seen as opacity (maximum grade 1) and epithelial damage (maximum 25 % of the corneal area) in one animal, which had completely resolved within 72 hours. Iridial irritation (grade 1) was observed in all animals after 1 hour which recurred in one animal after 48 hours. Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 14 days in all animals. Due to brown staining by the test substance, scoring for conjunctival redness was prohibited in all animals after 1 hour and was partly prohibited in one animal upto 72 hours. Brown staining of the conjunctivae and of the fur on the head and paws, caused by the test substance, was noted during the observation period. Remnants of the test substance were present in the eyes of all animals on day 1 and on the outside of the eyelids of two animals between 1 and 72 hours.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report Date:
1999

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Description Brown powder
Test substance storage At room temperature in the dark
Stability under storage conditions Not indicated
Expiry date 29 August 2002 (allocated by NOTOX, 1 year after)

Test animals / tissue source

Species:
rabbit
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
As required by the Dutch Act on Animal Experimentation, the study protocol was reviewed and agreed by the Article 14-functionary and the Ethical Committee of NOTOX.
Species: Albino Rabbit, New Zealand White, (SPF-Quality)
Source: Charles River Nederland, the Netherlands
Age and body weight: at least 6 weeks old and body weights less than 3.5 kg
Identification: earmark
ENVIRONMENTAL CONDITIONS
Conditions: air-conditioned room with approximately 15 air changes per hour
Temperature: 21°C
Relative humidity of 50%.
Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity.
Lighting: 12 hours artificial fluorescent light and 12 hours dark per day.
Accomodation: Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
Acclimatisation period: at least 5 days before start of treatment under laboratory conditions.
Diet: Standard laboratory rabbit diet (LKK-20, pellet diameter 4mm, Hope Farms, Woerden, The Netherlands) approx. 100 gram per day. In addition, hay (BMI, Helmond, The Netherlands) was provided once a week.
Certificates of analysis were examined and then retained in the NOTOX archives.
Free access to tap-water diluted with decalcified water.
Certificates of quarterly analysis were examined and then retained in the NOTOX archives.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
instillation of 99.6 ± 0.3 mg of the test substance (a volume of approximately 0.1 ml)
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
1, 24, 48 and 72 hours and 7 and 14 days after instillation
Number of animals or in vitro replicates:
3 animals of one sex
Details on study design:
Since marked effects were considered possible this eye irritation study was started by treatment of a single rabbit (sentinel).
The two other animals were treated in a similar manner 14 days later, after considering the degree of eye irritation observed in the first animal except that the eyes of the third rabbit were anaesthesised with a locai anaesteticum (0.4 % Benoxinate Hydroxide, Chravlls Pharmaceuticals, England), approximately 8 minutes prior to instillation.
A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the eyes, which were free from any abnormality.
Each animal was treated by instillation in the conjunctival sac of one of the eyes 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.
Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. When considered necessary, this procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.
OBSERVATION
Mortality/Viability: twice daily
Toxicity: once a day
Body weights: day of the treatment
Irritation: The eyes of each animal were examined approximately 1, 24, 48 and 72 hours and 7 and 14 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.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Remarks:
opacity
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
0.1
Max. score:
1
Reversibility:
fully reversible
Remarks on result:
other: brown coloration
Irritation parameter:
iris score
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
0.1
Max. score:
2
Reversibility:
not fully reversible within: 48 hours
Remarks on result:
other: brown coloration
Irritation parameter:
conjunctivae score
Remarks:
redness
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
2.9
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks on result:
other: brown coloration
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
3 animals
Time point:
other: 24, 72 hours
Score:
1.2
Max. score:
4
Reversibility:
not fully reversible within: 14 days
Remarks on result:
other: brown coloration
Irritant / corrosive response data:
Vocalisation of two rabbits was noted immediately after instillation, indicating pain.
Therefore, the eyes of the third rabbit were anaesthesised with a local anaesteticum (0.4 % Benoxinate Hydroxide, Chravils Pharmaceuticals, England), approximately 8 minutes prior to instillation.
Irritation
Instillation of approximately 99.6 mg of the test substance (a volume of approximately 0.1 ml) into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. Corneal injury was seen as opacity (maximum grade 1) and epithelial damage (maximum 25 % of the corneal area) in one animal, which had completely resolved within 72 hours.
Iridial irritation (grade 1) was observed in all animals after 1 hour which recurred in one animal after 48 hours.
Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 14 days in all animals.
Due to brown staining by the test substance, scoring for conjunctival redness was prohibited in all animals after 1 hour and was partly prohibited in one animalup to 72 hours.
Corrosion
There was no evidence of ocular corrosion.
Other effects:
Colouration
Brown staining of the conjunctivae and of the fur on the head and paws, caused by the test substance, was noted during the observation period. Remnants of the test substance were present in the eyes of all animals on day 1 and on the outside of the eyelids of two animals between 1 and 72 hours.
Toxicity/Mortality
No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Classification criteria according to the CLP Regulation 1272/2008 and its amendments
Conclusions:
Eye irritation was caused by the instillation of the substance.
In addition, brown staining of the conjunctivae and of the fur on the head and paws was noted during the observation period.
Executive summary:

Acute eye irritation/corrosion study was carried out based on the guidelines OECD No.405 "Acute Eye Irritation/Corrosionand the EU B.5, "Acute Toxicity - Eye irritation".

Single samples of approximately 99.6 mg of the similar substance (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours and 7 and 14 days after instillation. 

Vocalisation of two rabbits was noted immediately after instillation, indicating pain. Therefore, the eyes of the third rabbit were anaesthesised with a local anaestheticum. Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae. Corneal injury was seen as opacity (maximum grade 1) and epithelial damage (maximum 25 % of the corneal area) in one animal, which had completely resolved within 72 hours. Iridial irritation (grade 1) was observed in all animals after 1 hour which recurred in one animal after 48 hours. Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 14 days in all animals. Due to brown staining by the test substance, scoring for conjunctival redness was prohibited in all animals after 1 hour and was partly prohibited in one animal upto 72 hours. Brown staining of the conjunctivae and of the fur on the head and paws, caused by the test substance, was noted during the observation period. Remnants of the test substance were present in the eyes of all animals on day 1 and on the outside of the eyelids of two animals between 1 and 72 hours.