Reaction mass of 1,4-bis(methylamino)anthraquinone and 1,4-bis[(2-ethylhexyl)amino]anthraquinone and 1-[(2-ethylhexyl)amino]-4-(methylamino)anthraquinone and 9,10-Anthracenedione, 1,4-bis(pentylamino)-, branched and linear and 9,10-Anthracenedione, 1-(methylamino)-4-(pentylamino)-, branched and linear and 9,10-Anthracenedione, 1-[(2-ethylhexyl)amino]-4-(pentylamino)-, branched and linear

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5-5-1982 to 5-12-1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: OECD guideline study performed in accordance with GLP; exact details of test material (certificate of analysis, Characterisation) are not included in the report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD Guidelines for Testing of Chemicals, 1981

Deviations:

not specified

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 1,4-bis(methylamino)anthraquinone and 1,4-bis[(2-ethylhexyl)amino]anthraquinone and 1-[(2-ethylhexyl)amino]-4-(methylamino)anthraquinone and 9,10-Anthracenedione, 1,4-bis(pentylamino)-, branched and linear and 9,10-Anthracenedione, 1-(methylamino)-4-(pentylamino)-, branched and linear and 9,10-Anthracenedione, 1-[(2-ethylhexyl)amino]-4-(pentylamino)-, branched and linear

EC Number:

911-360-1

Molecular formula:

variable structures

IUPAC Name:

Reaction mass of 1,4-bis(methylamino)anthraquinone and 1,4-bis[(2-ethylhexyl)amino]anthraquinone and 1-[(2-ethylhexyl)amino]-4-(methylamino)anthraquinone and 9,10-Anthracenedione, 1,4-bis(pentylamino)-, branched and linear and 9,10-Anthracenedione, 1-(methylamino)-4-(pentylamino)-, branched and linear and 9,10-Anthracenedione, 1-[(2-ethylhexyl)amino]-4-(pentylamino)-, branched and linear

Test material form:

not specified

Details on test material:

- Name of test material (as cited in study report): AUTOMATE BLUE 8 NONVOLATILE
- Lot/batch No.: 82-0268

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Animals and Animal Care:
Six young adult New Zealand White rabbits, weighing approximately 2.0 - 4.0 kg initially, were purchased from New York State Rabbit Development, Hartwick, NY (a USDA approved supplier) for use in this study. The rabbits were housed individually in wire mesh bottom cages in an environmentally monitored room. Animals were identified by use of ear tags and cage cards. NIH Animal Feed A (certified) and water were provided ad libitum. The animals were acclimated a minimum of 5 days. During this acclimation period, the rabbits were examined with respect to their general health to assure their suitability as test animals.

Housing: Individually housed in wire mesh-bottom cages
Food: NIH Animal Feed A, certified feed, ad libitum, supplied fresh daily. No known contaminants are expected to be present in the basal diet that would interfere with the conduct of this study.
Water: Tap water: Ad libitum. Water is monitored for contaminants at periodic intervals according to FDRL Standard Operating Procedures.
Environment: All animals will be housed in environment-controlled rooms with temperature and relative humidity regulated as per "Guide for the Care and Use of Laboratory Animals". Filtered air will be supplied to provide 12-15 air changes per hour. A 12 hour light/dark cycle will be maintained.
Quarantine: Minimum of 5 days. During this conditioning period, the rabbits will be observed for any clinical signs of disease. All rabbits with any evidence of disease or physical abnormalities will be discarded.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: Untreated eye served as control.

Amount / concentration applied:

A volume of 0.1 ml of the test article (undiluted) was instilled into one eye of each rabbit.

Duration of treatment / exposure:

Single exposure. Eyes are not irrigated after test article installation.

Observation period (in vivo):

The eyes were examined after treatment using fluorescein dye procedures and the grade of ocular reaction for each animal was recorded at 1, 24, 48 and 72 hours and again at 7 days using the Draize Scale for Scoring Ocular Lesions. If injury persisted the eyes were further examined and scored every 3 days for 13 days or until all signs of reversible irritation subsided.

Number of animals or in vitro replicates:

6

Details on study design:

The eyes were examined after treatment using fluorescein dye procedures and the grade of ocular reaction for each animal was recorded at 1, 24, 48 and 72 hours and again at 7 days using the Draize Scale for Scoring Ocular Lesions.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

other: Mean score for all animals

Time point:

other: 24, 48 and 72 h

Score:

0.5

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

other: Mean score for all animals

Time point:

other: 24, 48 and 72 h

Score:

0.33

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Basis:

other: Mean score for all animals

Time point:

other: 24, 48 and 72 h

Score:

0.5

Max. score:

3

Results 4

Irritation parameter:

chemosis score

Basis:

other: Mean score for all animals

Time point:

other: 24, 48 and 72 h

Score:

0.56

Max. score:

4

Irritant / corrosive response data:

See results section.

Any other information on results incl. tables

Individual Rabbit Eye Irritation Scores

			Reading
Rabbit Number	Item	Tissue	1 hr	24 hr	48 hr	72 hr	Day 7
0896	A	Cornea Opacity	0	1	1	1	0
	B	Cornea Area	0	1	1	1	0
	(1)	Cornea Total=(AxB)x5	0	5	5	5	0
	C	Iris	0	1	0	0	0
	(2)	Iris Total=(C)x5	0	5	0	0	0
	D	Conjunctiva Redness	0	2	1	0	0
	E	Conjunctiva Chemosis	0	2	0	0	0
	F	Conjunctiva Discharge	0	2	1	0	0
	(3)	Conjunctiva Total (D+E+F)x2	0	12	4	0	0
		Totals Added= (1+2+3)	0	22	9	5	0
0897	A	Cornea Opacity	0	1	1	1	0
	B	Cornea Area	0	3	3	3	0
	(1)	Cornea Total=(AxB)x5	0	15	15	15	0
	C	Iris	0	1	1	1	0
	(2)	Iris Total=(C)x5	0	5	5	5	0
	D	Conjunctiva Redness	0	2	1	0	0
	E	Conjunctiva Chemosis	0	2	1	0	0
	F	Conjunctiva Discharge	0	1	1	0	0
	(3)	Conjunctiva Total (D+E+F)x2	0	10	6	0	0
		Totals Added= (1+2+3)	0	30	26	20	0
0898	A	Cornea Opacity	0	0	0	0	0
	B	Cornea Area	0	0	0	0	0
	(1)	Cornea Total=(AxB)x5	0	0	0	0	0
	C	Iris	0	0	0	0	0
	(2)	Iris Total=(C)x5	0	0	0	0	0
	D	Conjunctiva Redness	0	0	0	0	0
	E	Conjunctiva Chemosis	0	0	0	0	0
	F	Conjunctiva Discharge	0	0	0	0	0
	(3)	Conjunctiva Total (D+E+F)x2	0	0	0	0	0
		Totals Added= (1+2+3)	0	0	0	0	0
0901	A	Cornea Opacity	0	0	0	0	0
	B	Cornea Area	0	0	0	0	0
	(1)	Cornea Total=(AxB)x5	0	0	0	0	0
	C	Iris	0	0	0	0	0
	(2)	Iris Total=(C)x5	0	0	0	0	0
	D	Conjunctiva Redness	0	0	0	0	0
	E	Conjunctiva Chemosis	0	0	0	0	0
	F	Conjunctiva Discharge	0	0	0	0	0
	(3)	Conjunctiva Total (D+E+F)x2	0	0	0	0	0
		Totals Added= (1+2+3)	0	0	0	0	0
0902	A	Cornea Opacity	0	0	0	0	0
	B	Cornea Area	0	0	0	0	0
	(1)	Cornea Total=(AxB)x5	0	0	0	0	0
	C	Iris	0	0	0	0	0
	(2)	Iris Total=(C)x5	0	0	0	0	0
	D	Conjunctiva Redness	0	0	0	0	0
	E	Conjunctiva Chemosis	0	0	0	0	0
	F	Conjunctiva Discharge	0	0	0	0	0
	(3)	Conjunctiva Total (D+E+F)x2	0	0	0	0	0
		Totals Added= (1+2+3)	0	0	0	0	0
0904	A	Cornea Opacity	0	1	1	1	0
	B	Cornea Area	0	1	1	1	0
	(1)	Cornea Total=(AxB)x5	0	5	5	5	0
	C	Iris	0	1	1	0	0
	(2)	Iris Total=(C)x5	0	5	5	0	0
	D	Conjunctiva Redness	0	2	1	0	0
	E	Conjunctiva Chemosis	0	2	2	1	0
	F	Conjunctiva Discharge	0	2	2	1	0
	(3)	Conjunctiva Total (D+E+F)x2	0	12	10	4	0
		Totals Added= (1+2+3)	0	22	20	9	0

Initial Body Weights:

Animal Number              Initial Body Weight (kg)

82 -0896                            3.01

82 -0897                            3.05

82 -0898                            3.00

82 -0901                            2.97

82 -0902                            3.15

82 -0904                            2.72

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Three of six animals were noted with positive cornea opacity effects (maximum score grade 1), iridal effects (maximum score grade 1), conjunctival redness (maximum score grade 2) and conjunctival chemosis (maximum score grade 2). The other three animals showed no effects throughout the study. All positive effects observed had resolved by day 7. The individual animal mean scores for the 24, 48 and 72 hr time-point indicated that this material would not be classified under EU regulations.

Executive summary:

The primary ocular irritation potential of the test material was evaluated in this study with New Zealand White rabbits. One group of six albino rabbits received a single, unwashed exposure. A 0.1- ml dose of the test article was instilled into the lower conjunctival sac of one eye. The eyelids were held together for approximately one second and released. The other eye served as an untreated control. The eyes were examined for ocular reactions in accordance with the method of Draize at approximately 1, 24, 48 and 72 hours after dosing and on study day 7. Sodium fluorescein dye was used to aid in revealing possible effects.

Three of six animals were noted with positive cornea opacity effects (maximum score grade 1), iridal effects (maximum score grade 1), conjunctival redness (maximum score grade 2) and conjunctival chemosis (maximum score grade 2). The other three animals showed no effects throughout the study. All positive effects observed had resolved by day 7. The individual animal mean scores for the 24, 48 and 72 hr time-point indicated that this material would not be classified under EU regulations.