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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:
supporting study
Study period:
9 May 2012 to 28 May 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP 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.

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
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
944127-07-1
IUPAC Name:
944127-07-1
Test material form:
other: solid (unspecified)
Details on test material:
- Appearance: White Solid
- Storage conditions of the test material: Ambient (18 to 36 °C)
- pH of test material: 4.60

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Age at study initiation: 6 to 7 months
- Weight at study initiation: 2.70 to 2.75 kg
- Fasting period before study: No
- Housing: The rabbits were housed individually in rabbit cages (size: approximately 65 cm long x 65 cm deep x 45 cm high) with a noryl shallow cage body and facilities for pelleted feed and drinking water. The litter collection trays (noryl waste tray) were changed daily (except on Sundays). A noryl perforated raised shelf was provided to the animals for enrichment.
- Diet (e.g. ad libitum): Rabbit feed was provided ad libitum in a stainless steel feed hopper.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in a water filter-cum-purifier was provided to animals ad libitum in 750 mL bottles with sipper tubes. The water in the bottles was replenished once daily and the water bottles were changed once a week.
- Acclimation period: Five to twelve days under laboratory conditions.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 22 °C
- Humidity (%): 56 to 66 % (relative)
- Air changes (per hr): 12 to 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours of light and 12 hours of dark

IN-LIFE DATES: From: 9 May 2012 To: 28 May 2012

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The right eye of each rabbit remained untreated and served as the reference control.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g
Duration of treatment / exposure:
The eye lids were gently held together for about one second, in order to minimise loss of the test material.
Observation period (in vivo):
7 days
Number of animals or in vitro replicates:
3 male animals
Details on study design:
TEST MATERIAL INSTALLATION
- Eye examinations: Prior to instillation, both eyes of each rabbit were examined with the aid of a pen light for irritation and ocular defects. The eyes were then checked for opacity using ophthalmic fluorescein sodium strips. The eyes were then irrigated with enough physiological saline (0.9 % NaCl) to remove any excess fluorescein (approximately 30 seconds). Using an ultraviolet lamp, the eyes were checked and evaluated for pre-existing corneal damage. No corneal defects were observed in any of the animals.
The finely ground test material was placed in the everted lower lid (conjunctival sac) of the left eye of each of three rabbits in a stepwise manner.
- Stepwise exposure of animals: A single rabbit was initially exposed to the test material. Prior to instillation, a local anaesthetic (4 % Lidocaine) was used in both the test and control eye. No corrosive effect or severe irritation was observed and all scores were 0 at 7 days post-installation; hence the test was completed using two additional animals.

SCORING SYSTEM: The eyes were evaluated with the illumination of a white light source at intervals of approximately 1, 24, 48 and 72 hours and 4 and 7 days post instillation of the test material according to the "Scale for Scoring Ocular Lesions" (Draize, 1977).

TOOL USED TO ASSESS SCORE: At 24, 48 and 72 hours and 4 and 7 days post installation, the eyes were checked for opacity using ophthalmic fluorescein sodium strips. The eyes were then irrigated with enough physiological saline (0.9 % NaCl) to remove any excess fluorescein (approximately 30 seconds). Using an ultraviolet lamp, the eyes were evaluated to verify the extent or absence of corneal damage.

OBSERVATIONS
- Clinical Observations: The rabbits were observed for clinical signs of toxicity and mortality 4 times (at hourly intervals) on the day of test material instillation and once daily thereafter until the end of the observation period.
- Body Weights: Body weights were recorded at the start of acclimatisation, day 1 of treatment (just before test material instillation) and at termination of observation.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
mean
Remarks:
of 3 animals
Time point:
other: Mean score at 24, 48 and 72 hours
Score:
1.56
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: Mean score at 24, 48 and 72 hours
Score:
1.22
Max. score:
4
Reversibility:
fully reversible within: 4 days
Irritation parameter:
other: discharge
Basis:
mean
Remarks:
of 3 animals
Time point:
other: Mean score at 24, 48 and 72 hours
Score:
1.33
Max. score:
3
Reversibility:
fully reversible within: 4 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: Mean score at 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
other: The iris score for all animals at all time points was 0
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
of 3 animals
Time point:
other: Mean score at 24, 48 and 72 hours
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
The individual eye irritation scores are presented in Table 1.
There was no iritis observed in any treated eye during the study. At 1 hour post-instillation, conjunctival irritation was noted in the treated eye of all three rabbits.
Corneal opacity was observed in the treated eye of all three rabbits at 24 hours. All animals were free of corneal opacity and ocular irritation by day 7 (study termination).
Other effects:
There were no clinical signs of toxicity or mortality observed during the study.
The body weights of all rabbits increased slightly throughout the observation period. See Table 2.

Any other information on results incl. tables

Table 1: Individual Eye Irritation Scores

Rabbit Number

Time Point

Redness

Chemosis

Discharge

Iris

Cornea

RB9612

1 hour

1

2

2

0

0

24 hours

2

1

2

0

1

48 hours

2

1

1

0

1

72 hours

1

1

1

0

1

4 days

1

0

0

0

1

7 days

0

0

0

0

0

RB9613

1 hour

1

2

2

0

0

24 hours

2

2

2

0

1

48 hours

2

1

1

0

1

72 hours

1

1

1

0

1

4 days

1

0

0

0

0

7 days

0

0

0

0

0

RB9614

1 hour

2

2

2

0

0

24 hours

2

2

2

0

1

48 hours

1

1

1

0

1

72 hours

1

1

1

0

1

4 days

1

0

0

0

1

7 days

0

0

0

0

0

Table 2: Body Weight

Rabbit Number

Body weight (kg)

At start of acclimatisation

At treatment

At termination of observation

RB9612

2.58

2.70

2.77

RB9613

2.69

2.75

2.89

RB9614

2.68

2.74

2.90

Applicant's summary and conclusion

Interpretation of results:
other: Category 2
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study, the test material caused conjunctival irritation and corneal opacity, which cleared within 7 days. In accordance with EU criteria, the test material requires classification as a Category 2 eye irritant.
Executive summary:

The potential of the test material to cause eye irritation in male New Zealand White rabbits was assessed in accordance with the standardised guidelines OECD 405 and US EPA OPPTS 870.2400 under GLP conditions.

The test was performed in a stepwise manner. A single rabbit was initially exposed to the test material. Since the test material was not corrosive or severely irritating the study was completed using two additional rabbits.

0.1 g of the finely ground test material was placed in the everted lower lid (conjunctival sac) of the left eye of each of three rabbits. The lids were gently held together for about one second, in order to minimise loss of the test material. The right eye of each rabbit remained untreated and served as the reference control. The eyes were evaluated at 1, 24, 48 and 72 hours and 4 and 7 days post instillation according to the Draize scale for scoring ocular lesions.

There was no iritis observed in any treated eye during the study. At 1 hour post-instillation, conjunctival irritation was noted in the treated eye of all three rabbits. Corneal opacity was observed in the treated eye of all three rabbits at 24 hours. All animals were free of corneal opacity and ocular irritation by day 7 (study termination).

There were no clinical signs of toxicity observed, and none of the animals died, during the study. All rabbits appeared healthy and gained body weight throughout the observation period.

Under the conditions of this study, the test material caused conjunctival irritation and corneal opacity, which cleared within 7 days. In accordance with EU criteria, the test material requires classification as a Category 2 eye irritant on the basis of the average score for corneal opacity observed at 24, 48 and 72 hours being 1 in 3 out of 3 animals.