[No public or meaningful name is available]

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Study initiation date: 05 December 2012; Experimental completion date: 31 December 2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP Guideline study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Test material form:

other: solid

Details on test material:

- Name of test material (as cited in study report): SP4834
- Physical state: Light yellow solid
- Lot/batch No.: TS 12007
- Expiration date of the lot/batch: 01 April 2014
- Storage condition of test material: stored at room temperature (18-24ºC), under a nitrogen blanket.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Male New Zealand White albino rabbits were received from Covance Research Products, Inc., Denver, PA
- Age at study initiation: The selected animals were approximately 7 months old.
- Weight at study initiation: 3118.1 g to 4061.0 g
- Fasting period before study: No fasting period.
- Housing: all animals were housed individually in clean, stainless steel cages. Enrichment devices were provided to all animals as appropriate throughout the study for environmental enrichment. One leaf of fresh kale was offered to each rabbit every Monday, Wednesday, and Friday beginning after the day of receipt.
- Diet (e.g. ad libitum): Basal diet (certified feed) provided at approximately 150 g/day was provided ad libitum
- Water (e.g. ad libitum): municipal water, delivered by an automatic watering system, was provided ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Set to maintain conditions of 19 ± 3°C
- Humidity (%): Set to maintain conditions of 50 ± 20%
- Air changes (per hr): 10 fresh air changes per hour
- Photoperiod (hrs dark / hrs light): Fluorescent lighting provided illumination for a 12-hour light (0600 hours to 1800 hours)/12-hour dark photoperiod.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100-mg dose of the test substance was instilled into the lower conjunctival sac of the right eye.

Duration of treatment / exposure:

Single application.

Observation period (in vivo):

14 days.

Number of animals or in vitro replicates:

3.

Details on study design:

Method of test substance administration:
The test substance was placed directly into the cupped lower conjunctival sac of each rabbit’s right (test) eye. The eyelid was held closed for approximately 1 second after instillation and then released. The left eye of each animal was manipulated in an identical manner to simulate the dosing of the right eye.

Initially, a single animal was dosed to evaluate the ocular irritative potential of the test substance. This single rabbit received a single, unwashed exposure. No severe ocular damage was observed in this animal. Based on these results, and to complete the study, 2 additional rabbits each received a single, unwashed exposure of the test substance.

Parameters evaluated:
Mortality:
The rabbits were observed twice daily, once in the morning and once in the afternoon, for mortality and moribundity.

Ocular observations:
Both eyes of all rabbits were examined for ocular abnormalities prior to initiation of dosing. The pre-initiation examination included the use of sodium fluorescein and a direct ophthalmoscope for detection of corneal abnormalities. Only rabbits with no pre-existing ocular abnormalities were assigned to study.

Both eyes of all rabbits were examined macroscopically for ocular irritation in accordance with the method of Draize scoring criteria for ocular reactions, at approximately 1, 24, 48, and 72 hours after dosing and on study days 4, 7, 10, and 14, if irritation persisted. A direct ophthalmoscope was used during these observations to examine the corneal tissue. In addition, both eyes were further examined at 24 hours and at all subsequent observations with sodium fluorescein.

Body Weights:
Body weights were obtained and recorded on study day 0 (initiation) and at each rabbit’s termination from the study.

Termination:
After study termination, the rabbits were euthanized by intravenous injection of sodium pentobarbital and discarded.

Results and discussion

In vivo

Results

Irritation parameter:

overall irritation score

Basis:

other: Draize Score

Time point:

other: 24 hour

Score:

16

Max. score:

110

Irritant / corrosive response data:

Positive corneal irritation (grade 1) and iridial irritation (grade 1) were noted for 1 male (No. 9853), and positive conjunctival irritation (grade 2) was noted for the remaining 2 males (Nos. 9850 and 9851). Positive corneal and iridial irritation subsided by 24 hours post-instillation. Positive scores for conjunctival irritation subsided by 72 hours post-instillation. All ocular irritation subsided by study day 14.

Individual and average ocular irritation scores for treated eyes are presented in Table 1, and individual animal results (other findings), including sodium fluorescein examination results, are presented in Table 2 (see attached background material for Tables 1 and 2).

The left (control) eyes were free of evidence of ocular irritation and other findings for the duration of the study.

The Draize scale for scoring ocular irritation and method of score calculation are presented in attached background material.

Other effects:

Mortality:
There were no deaths during the study.

Body Weights:
There were no remarkable body weight changes noted during the study.

Any other information on results incl. tables

Ocular Irritation Results Summary:

Number of Animals with Positive Effect/Number of Animals Treated

Group	Cornea	Iris	Conjunctiva	Total	24 Hour Draize
100 mg/right eye, unwashed	1/3	1/3	2/3	3/3	16/110

 

Applicant's summary and conclusion

Conclusions:

The 24-hour post-instillation Draize score for SP4834 was 16/110. All ocular irritation subsided by study day 14.

Executive summary:

Objective:

The objective of this study was to determine the primary ocular irritative potential of the test substance in albino rabbits.

Test Guidelines:

The protocol was designed to be in general compliance with the OECD Guidelines for Testing of Chemicals, Section 405 (2002).

Study Design:

Initially, a single animal was dosed to evaluate the ocular irritative potential of the test substance. This single rabbit received a single, unwashed exposure. No severe ocular damage was observed in this animal. Based on these results, and to complete the study, 2 additional rabbits received single, unwashed exposures of the test substance. Each 100-mg dose of the test substance was instilled into the lower conjunctival sac of the right eye. The eyelid was held closed for approximately 1 second and released. The left eye of each animal was manipulated in an identical manner to simulate the dosing of the right eye and served as a contralateral control.

Results:

There were no deaths or remarkable body weight changes during the study.

Positive corneal irritation (grade 1) and iridial irritation (grade 1) were noted for 1 animal and conjunctival irritation (grade 2) was noted for the remaining 2 animals. Positive corneal and iridial irritation subsided by 24 hours post-instillation. Positive conjunctival irritation subsided by 72 hours post-instillation. All ocular irritation subsided by study day 14.

Conclusion:

The 24-hour post-instillation Draize score for SP4834 was 16/110. All ocular irritation subsided by study day 14. The following table presents the individual ocular irritation scores at 24, 48, and 72 hours post-instillation:

Animal	Tissue	24 Hours	48 Hours	72 Hours
9853	Cornea (O-A)	0 0	0 0	0 0
	Iris	0	0	0
	Conjunctive  (R-C-D)	1 1 0	1 0 0	0 0 0
9850	Cornea (O-A)	0 0	0 0	0 0
	Iris	0	0	0
	Conjunctive  (R-C-D)	2 1 0	1 1 0	1 0 0
9851	Cornea (O-A)	0 0	0 0	0 0
	Iris	0	0	0
	Conjunctive  (R-C-D)	2 1 0	2 1 0	1 0 0