N-[3-(dimethylamino)propyl] C6-9 alkyl amides

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From January 26, 2009 to February 03, 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

1044764-00-2

Molecular formula:

RCONHCH2CH2CH2N(CH3)2

Test material form:

liquid

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Animals
Number of Animals: 3
Sex: Females, nulliparous and non-pregnant
Species/Strain: Rabbit/New Zealand albino
Age: Young adult
Source: Received from Robinson Services, Inc. Clemmons, NC on January 7, 2009
Housing: The animals were singly housed in suspended stainless steel caging with mesh floors which conform to the size recommendations in the most recent Guide for the Care and Use of Laboratory Animals DHEW (NIH). Litter paper was placed beneath the cage and was changed at least three times per week.
Animal Room Temperature and Relative Humidity Ranges: 19-21ºC and 25-59%, respectively. The low humidity reading was due to a temporary malfunction of the environmental control system.
Photoperiod: 12-hour light/dark cycle
Acclimation Period: 19 days
Food: Pelleted Purina Rabbit Chow #5326
Water: Filtered tap water was supplied ad libitum by an automatic water dispensing system.
Contaminants: There were no known contaminants reasonably expected to be found in the food or water at levels which would have interfered with the results of this study.
Analyses of the food and water are conducted regularly and the records are kept at test fecility.
Cage: Each cage was identified with a cage card indicating at least the study number and identification and sex of the animal.
Animal: A number was allocated to each rabbit on receipt and a stainless steel ear tag bearing this number was attached to the animal.

Test system

Vehicle:

not specified

Controls:

yes, concurrent no treatment

Amount / concentration applied:

One-tenth of a milliliter of the test substance was instilled into the conjunctival sac of the right eye of each rabbit.

Observation period (in vivo):

Ocular irritation was evaluated at 1, 24, 48, and 72 hours and at 4 and 7 days post-instillation.

Number of animals or in vitro replicates:

3

Details on study design:

Preparation and Selection of Animals
Prior to test initiation, both eyes of a group of animals were examined using a white light source and a fluorescein dye procedure. One drop of 2% ophthalmic fluorescein sodium was instilled into both eyes of each rabbit. The eyes were rinsed with physiological saline (0.9% NaCl) approximately 30 seconds after instillation of the fluorescein and then evaluated for corneal damage using an ultraviolet light source. Prior to instillation, the eyes were re-examined and scored for abnormalities according to the "Scale for Scoring Ocular Lesions". Three healthy naive animals (not previously tested) without pre-existing ocular irritation were selected for test.

Instillation
Prior to instillation, 2-3 drops of ocular anesthetic (Tetracaine Hydrochloride Ophthalmic Solution, 0.5%) were placed into both the treated and control eye of each animal. One-tenth of a milliliter of the test substance was then instilled into the conjunctival sac of the right eye of each rabbit by pulling the lower lid away from the eyeball. The upper and lower lids were then gently held together for about one second before releasing to minimize loss of the test substance. The other eye of each rabbit remained untreated with the test substance and served as a control. The rabbits were then returned to their designated cages.

Ocular Scoring
Ocular irritation was evaluated using a high-intensity white light (Mag Lite) in accordance with Draize et al. at 1, 24, 48, and 72 hours and at 4 and 7 days post-instillation. The fluorescein dye evaluation was done on the treated eye at 24 hours and as needed at subsequent scoring intervals to evaluate the extent of corneal damage or to verify reversal of effects. Individual scores were recorded for each animal. In addition to observations of the cornea, iris and conjunctivae, any other observed lesions were noted. The average score for all rabbits at each scoring period was calculated to aid in data interpretation.

Classification of Eye Scores
The time interval with the highest mean score (Maximum Mean Total Score - MMTS) for all rabbits was used to classify the test substance by the system of Kay and Calandra. The average individually-determined irritation scores for cornea, iris and conjunctiva (redness and chemosis) across 24, 48 and 72 hours were calculated for EEC classification.

Cage-Side Observations
The animals were observed for signs of gross toxicity and behavioral changes at least once daily during the test period. Observations included gross evaluation of skin and fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity and behavior pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhea, and coma.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

not reversible

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

not reversible

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

not reversible

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.7

Max. score:

2

Reversibility:

not reversible

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not reversible

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.7

Max. score:

2

Reversibility:

not reversible

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.7

Max. score:

3

Reversibility:

not reversible

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not reversible

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.3

Max. score:

3

Reversibility:

not reversible

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

not reversible

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

3

Max. score:

4

Reversibility:

not reversible

Irritant / corrosive response data:

All animals appeared active and healthy during the study. Apart from the eye irritation there were no other signs of gross toxicity, adverse pharmacologic effects, or abnormal behavior.
One hour after test substance instillation, all three treated eyes exhibited corneal opacity, iritis and ‘positive’ conjunctivitis. The overall incidence and severity of irritation appeared to be severe. A brownish red discharge and pannus was noted for all three treated eyes between Days 1 and 7.
Therefore following the Day 7 evaluation, the study was terminated and all animals were euthanized for humane reasons. The maximum mean total score for the test substance is 49.0.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Under the study conditions, the test substancewas severely irritating to the eye.

Executive summary:

A study was conducted to determine the eye irritation potential of the test substance in female New Zealand albino rabbits according to OECD Guideline 405, EU Method B.5 and EPA OPPTS Method 870.2400, in compliance with GLP. A dose of 0.1 mL of the test substance was administered into the conjunctival sac of the right eye of the rabbits. The upper and lower lids were then gently held together for about one second in order to prevent loss of test substance. The left eye, which remained untreated, served as a control. Ocular irritation was evaluated using a high-intensity white light (Mag Lite) in accordance with Draize et al.1 at 1, 24, 48, and 72 h and at 4 and 7 d post-instillation. The fluorescein dye evaluation was done on the treated eye at 24 h and as needed at subsequent scoring intervals to evaluate the extent of corneal damage or to verify reversal of effects. All animals appeared active and healthy during the study. Apart from the eye irritation there were no other signs of gross toxicity, adverse pharmacologic effects or abnormal behavior. One hour after test substance instillation, all three treated eyes exhibited corneal opacity, iritis and ‘positive’ conjunctivitis. The overall incidence and severity of irritation appeared to be severe. A brownish red discharge and pannus was noted for all three treated eyes between Days 1 and 7. Therefore, following the Day 7 evaluation, the study was terminated and all animals were euthanized for humane reasons. The maximum mean total score for the test substance was 49.0. Under the study conditions, the substance was severely irritating to the eye (Oley, 2009).