Registration Dossier

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:
2011-11-15 to 2011-11-22
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Study conducted in compliance 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 and done to a valid guideline.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
On 11 occasions during the study the recorded humidity was outside the target range (30-70%). The actual humidity range was 28-50 %. This deviation is considered to have no impact on the outcome of the study or the interpretation of the results.
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
EC No 440/2008
Deviations:
yes
Remarks:
On 11 occasions during the study the recorded humidity was outside the target range (30-70%). The actual humidity range was 28-50 %. This deviation is considered to have no impact on the outcome of the study or the interpretation of the results.
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
yes
Remarks:
On 11 occasions during the study the recorded humidity was outside the target range (30-70%). The actual humidity range was 28-50 %. This deviation is considered to have no impact on the outcome of the study or the interpretation of the results.
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Dark Brown Liquid
Specific details on test material used for the study:
EC Number: 939-946-2

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
Justification of strain: The New Zealand White rabbit is one of the standard strains used for acute irritation toxicity studies.
Number of animals: 3 animals
Sex: Male
Age of animals: ~11 weeks
Body weight range at dosing: 2915 – 2954 g
Body weight range at the end: 3108 – 3181 g
Husbandry
Animal health: Only healthy animals were used for the study, as certified by the veterinarian.
Acclimation period: 6 days
Light: 12 hours of light/12 hours of dark
Temperature range during the study: 18.3 – 20.1 °C
Relative humidity during the study: 28 – 50 %
Housing/Enrichment: Rabbits were individually housed in AAALAC approved metal wire rabbit cages. The cages were of an open wire structure and were placed together to allow some social interaction with rabbit(s) in adjoining cages.
Ventilation: 15-20 air exchanges/hour.
The temperature and relative humidity was recorded twice every day during the acclimatisation and experimental phases.
Food and feeding
The animals received Purina Base – Lap gr. diet for rabbit. The detailed description of the contents of the lots used are archived with the raw data. The food was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
Water supply and quality control
Tap water from the municipal supply was supplied ad libitum, from an automatic system. The water was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study and was considered to be fit for human consumption.
The quality control analysis is performed once every 3 months and microbiological assessment is performed monthly.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: the untreated eye
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
1 week
Observation period (in vivo):
1, 24, 48 and 72 hours and 1 week
Number of animals or in vitro replicates:
3
Details on study design:
Approximately 1 hour before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect using a hand-held slit-lamp. Only animals free of ocular damage were used. The eyes were not anaesthetised prior to instillation of the test material.
Initially, a single rabbit was treated. A volume of 0.1 mL of the test material was placed into the conjunctival sac of the left eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about 1 second immediately after treatment, to prevent loss of the test material, and then released. The right eye remained untreated and was used for control purposes. Immediately after administration of the test material, an assessment of the initial pain reaction was made according to the 6 point scale.
Following a review of the ocular responses produced in the first treated animal, 2 additional animals were treated.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 1 hour
Score:
16
Max. score:
110
Reversibility:
fully reversible within: 1 week
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 24 hours
Score:
9.33
Max. score:
110
Reversibility:
fully reversible within: 1 week
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 48 hours
Score:
6.67
Max. score:
110
Reversibility:
fully reversible within: 1 week
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 72 hours
Score:
5.33
Max. score:
110
Reversibility:
fully reversible within: 1 week
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: 1 week
Score:
0
Max. score:
110
Irritant / corrosive response data:
Observations
Assessment of ocular damage/irritation was made approximately 1, 24, 48 and 72 hours then 1 week following treatment, according to the numerical evaluation given by Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49).
The eyes were further examined using 2% fluorescein solution at approximately 24 hours before treatment and then 24, 48, 72 hours and 1 week after treatment. One drop of 2% sodium fluorescein was applied to the corneal surface for approximately 30 seconds, then rinsed with physiological saline solution. Examination was performed with the use of a hand slit-lamp and recorded as either the presence of staining or no staining.
The duration of the observation period was sufficient to identify reversibility of changes. Any clinical signs of toxicity or signs of ill-health during the study were recorded. Rabbits were weighed on day 0 (prior to treatment) and on day 7 (prior to termination).
Other effects:
During the study, the control eye of all animals was symptom-free.
The bodyweights of all rabbits were considered to be within the normal range of variability throughout the study.
No clinical signs of systemic toxicity were observed in any animals in this study.
No mortality occurred in this study.

Any other information on results incl. tables

Ocular Reactions Initial Pain Reaction (IPR) scores were taken after the treatment of the test item and a score of 1 or 2 was observed in all animals.

One hour after the application, conjunctival chemosis (score 3), discharge (score 3) and conjunctival redness (score 2) were observed in all rabbits.

At 24 hours after treatment, conjunctival chemosis (score 2 or 1), discharge (score 2 or 1) and conjunctival redness (score 2) were seen in all rabbits.

At 48 hours after treatment conjunctival redness (score 2 and 1) was observed in all animals, discharge (score 2 and 1) in all animals and chemosis (score 2) was seen in one rabbit.

At 72 hours after treatment, conjunctival redness (score 2 and 1) was seen in all rabbits. Conjunctival chemosis (score 1) was seen in one animal. Conjunctival discharge (score 2 and 1) was observed in two animals.

At 1 week after treatment, no signs were observed and all animals were symptom free.

Applicant's summary and conclusion

Interpretation of results:
moderately irritating
Remarks:
Migrated information Criteria used for interpretation of results: other: Kay and Calandra
Conclusions:
The test substance was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to a modified Kay and Calandra classification system.
The study is considered to be relevant, reliable, adequate for risk assessment, and adequate for classification purposes.
Executive summary:

In a primary eye irritation study, 0.1mL of the test substance was instilled into the conjunctival sac of the left eye of each of 3 adult male New Zealand White rabbits. The untreated right eyes served as the control. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours and 1 week after test material instillation. Observations with fluorescein staining were made at approximately 24 hours before treatment and then 24, 48, 72 hours and 1week after treatment.

Results

Initial Pain Reaction (IPR) scores were taken after the treatment of the test item and a score of 1 or 2 was observed in the animals. Discharge was observed in all animals at the 1, 24 and 48 hour observations and in two animals, 72 hours after treatment. Conjunctival redness was observed in all 3 animals up until 72 hours after treatment. Chemosis was seen in all animals at the 1 and 24 hour observation which persisted in one animal until the 72 hour observation. No other signs were observed and all animals were symptom free 1 week after treatment. Fluorescein staining was negative in all animals during the experiment During the study, the control eye of all animals was symptom-free. No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred. The body weights of all rabbits were considered to be within the normal range of variability.

Conclusion

The test substance was classified as a moderate irritant (Class 5 on a 1 to 8 scale) to the rabbit eye according to the modified Kay and Calandra classification system.