Registration Dossier

Toxicological information

Eye irritation

Currently viewing:

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Animal Information
Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Envigo RMS (UK) Limited, Leicestershire, UK. At the start of the study the animals weighed 2.49 or 3.24 kg and were 12 to 52 weeks old. After an acclimatization period of at least 5 days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.
Animal Care and Husbandry
The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The left eye remained untreated and was used for control purposes.
Amount / concentration applied:
0.1 mL of the test item, which was found to weigh approximately 82 mg
Duration of treatment / exposure:
1 hour
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
2
Details on study design:
Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.
Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre dose anesthesia of ocular anesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.
A volume of 0.1 mL of the test item, which was found to weigh approximately 82 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Annex 1.
After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated.
Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation (Draize, J.H, 1977) given in Annex 2.
Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Any clinical signs of toxicity, if present, were also recorded.
An additional observation was made on Day 7 to assess the reversibility of the ocular effects.
Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
animal: 75328 Female
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
cornea opacity score
Basis:
animal: 75370 Male
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
0
Max. score:
4
Reversibility:
other: No effects observed
Irritation parameter:
iris score
Basis:
animal: 75328 Female
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 24 hours
Irritation parameter:
iris score
Basis:
animal: 75370 Male
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
0
Max. score:
2
Reversibility:
other: No effects observed
Irritation parameter:
other: redness
Basis:
animal: 75328 Female
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
other: redness
Basis:
animal: 75370 Male
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal: 75328 Female
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
animal: 75370 Male
Time point:
other: Highest score at 24, 48, 72 hours and 7 days
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
Individual and mean scores for ocular irritation are given in Appendix 1.
No corneal effects were noted during the study.
Iridial inflammation was noted in one treated eye 1 hour after treatment.
Moderate conjunctival irritation was noted in both treated eyes 1 and 24 hours after treatment. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye at the 48 Hour observation. Minimal conjunctival irritation was noted in both treated eyes at the 72 Hour observation.
Both treated eyes appeared normal at the 7 Day observation.
No corrosive effects were noted during the study. The test item did not induce significant or irreversible damage to the rabbit eye.
Other effects:
Body Weight
Individual body weights and body weight change are given in Appendix 2.
Both animals showed expected gain in body weight during the study.

Any other information on results incl. tables

Appendix 1     Eye Irritation Scores

Individual Scores for Eye Irritation

Rabbit Number and Sex

IPR

Evaluation interval

Corneal Opacity

Area of Corneal Opacity

Iris

Conjunctivae

Redness

Chemosis

Discharge

75328Female

0

1 Hour

0

0

1

2

1

2

75370Male

0

0

0

0

2

1

2

75328Female

 

24 Hour

0

0

0

2

1

1

75370Male

 

0

0

0

2

1

1

75328Female

 

48 Hour

0

0

0

2

1

1

75370Male

 

0

0

0

2

1

0

75328Female

 

72 Hour

0

0

0

1

0

0

75370Male

 

0

0

0

1

1

0

75328Female

 

7 Days

0

0

0

0

0

0

75370Male

 

0

0

0

0

0

0

IPR=Initial pain reaction

Appendix 1 (continued)       Eye Irritation Scores

Mean Values after 24, 48 and 72 Hours

Rabbit Number
and Sex

Number of available data points

Corneal Opacity

Iris

Conjunctivae

Redness

Chemosis

75328Female

3

0.0

0.0

1.7

0.7

75370Male

3

0.0

0.0

1.7

1.0

Assessment According to Regulation (EC) No. 1272/2008

Evaluated Intervals

Corneal Opacity

Iris

Conjunctivae

Redness

Chemosis

24 Hours

Not classified

Not classified

Not classified

Not classified

48 Hours

72 Hours

 

Appendix 2     Individual Body Weights and Body Weight Change

Rabbit Number
and Sex

Individual Body Weight (kg)

Body Weight Change (kg)

Day 0

Day 7

75328Female

3.24

3.30

0.06

75370Male

2.49

2.52

0.03

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the findings in this study, the test item does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.
Executive summary:

The primary eye irritation potential of the test item was investigated according to a method compatible with OECD test guideline No. 405 and Method B5. The test item was applied by instillation of 0.1 mL into the right eye of each of two young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48, 72 hours and 7 days after test item instillation. 

The mean score was calculated separately for each animal across three scoring times (24, 48 and 72 hours after instillation) for corneal opacity, iritis, redness and chemosis of the conjunctivae. The individual mean scores for corneal opacity and iritis was 0.0 for both animals. The individual mean scores for the conjunctivae were 1.7 for reddening for both animals and 0.7 or 1.0 for chemosis. 

The instillation of the test item into the eye resulted in conjunctival irritation. These effects were reversible and were no longer evident 7 days after treatment in both animals (end of the observation period). No abnormal findings were observed in the cornea of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

According to the findings in this study, the test item does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.