9,10-Anthracenedione, 1,4-bis[(2-ethylhexyl)amino]-5,8-dihydroxy-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD TG 405 and U.S. EPA Health Effects TG, OPPTS 870.2400 and in accordance with the Principles of Good Laboratory Practice (GLP).

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

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:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Principles of method if other than guideline:

not applicable

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,4-bis[(2-ethylhexyl)amino]-5,8-dihydroxy-9,10-dihydroanthracene-9,10-dione

EC Number:

807-717-5

Cas Number:

295800-70-3

Molecular formula:

C30H42N2O4

IUPAC Name:

1,4-bis[(2-ethylhexyl)amino]-5,8-dihydroxy-9,10-dihydroanthracene-9,10-dione

Test material form:

other: Blue solid

Details on test material:

- Name of test material (as cited in study report): Spectrace MD-810 Marker (Solvent Free)
- Physical state: Blue solid
- Lot/batch No.: Lot #RWS-2-43, TD#04-049
- Expiration date of the lot/batch: September 23, 2007
- Stability under test conditions: Test substance was expected to be stable for the duration of testing.
- Storage condition of test material: Stored at room tempertaure.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Robinson Services, Inc. Clemrnons, NC on November 17,2004
- Age at study initiation: Young adult.
- Weight at study initiation:
- 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 DHE W (IVIH). Litter paper was placed beneath the cage and was changed at least three times per week.
- Diet (e.g. ad libitum): Pelleted Purina Rabbit Chow #5326
- Water (e.g. ad libitum): Filtered tap water was supplied ad-libitum by an automatic water dispensing system.
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22°C
- Photoperiod (hrs dark / hrs light): 12-hour lightldark cycle

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

Prior to use, the test substance was ground to a fine powder with a mortar and pestle. One-tenth of a milliliter (0.03 grams) of the ground test substance was 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.

Duration of treatment / exposure:

72 hours

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance. 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

Number of animals or in vitro replicates:

2 male and 1 female

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: 72 hours

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: fluorescein

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 post-instillation. The fluorescein dye evaluation procedure was used at 24 hours to evaluate the extent of corneal damage or to verify reversal of effects.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

other: Not applicable

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 Hours

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 Hours

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 Hours

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 72 Hours

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.33

Max. score:

2

Reversibility:

fully reversible within: 72 Hours

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

fully reversible within: 72 Hours

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.67

Max. score:

1

Reversibility:

fully reversible within: 72 Hours

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 72 hours

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

1

Reversibility:

fully reversible within: 72 hours

Irritant / corrosive response data:

All animals appeared active and healthy. Apart from the eye irritation noted below, 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, intis, and conjunctivitis. The overall incidence and severity of irritation decreased with time. All animals were free of ocular irritation by 72 hours. The Maximum Mean Total Score of Spectracem MD-8 10 Marker (Solvent Free) is 21.3.

Other effects:

not applicable

Any other information on results incl. tables

None.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

One hour after test substance instillation, all three treated eyes exhibited corneal opacity, intis, and conjunctivitis. The overall incidence and severity of irritation decreased with time. All animals were free of ocular irritation by 72 hours. The Maximum Mean Total Score of Spectracem MD-8 10 Marker (Solvent Free) is 21.3.

Executive summary:

A primary eye irritation test was conducted with rabbits to determine the potential for Spectrace® MD-8 10 Marker

(Solvent Free) to produce irritation from a single instillation via the ocular route. Under the conditions of this study,

the observed ocular effects of Spectrace® MD-810 Marker (Solvent Free) were slight irritation to the eye.

Prior to its use in the study, the test substance was ground to a fine powder. One-tenth of a milliliter (0.03 grams) of

the ground test substance was instilled into the right eye of three healthy rabbits. The left eye remained untreated

and served as a control. Ocular irritation was evaluated by the method of Draize et al.1. One hour after test substance instillation, all three treated eyes exhibited corneal opacity, iritis, and conjunctivitis. The overall incidence and severity of irritation decreased with time. All animals were free of ocular irritation by 72hours.

The incidence, severity and reversibility of irritation are detailed below:

Time Post Instillation	Corneal Opacity	Iritis	Conjunctivitis
1 Hour	3/3	3/3	3/3
24 Hours	0/3	3/3	3/3
48 Hours	0/3	3/3	3/3
72 Hours	0/3	0/3	0/3

 

 

Time Post Instillation	Severity of Irritation- Mean Score
1 Hour	21.3
24 Hours	12.3
48 Hours	9.0
72 Hours	0