9,10-Anthracenedione, 1,4-diamino-, N,N'-mixed 2-ethylhexyl and 3-[(2-ethylhexyl)oxy]propyl and 3-methoxypropyl derivs.

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

5-5-1982 to 5-9-1982

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 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

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Animals and Animal Care:
Six adult New Zealand White rabbits, weighing between 2 and 4 kg initially, were purchased from New York State Rabbit Development, Hartwick, N.Y. (a USDA approved supplier) and used in this study. The rabbits were housed individually in suspended cages in an environmentally controlled room. During an acclimation period of no less than 5 days the animals were examined with respect to their general health to assure their suitability as test animals. NIH Animal Feed A (certified) and water were provided ad libitum. The animals were identified by use of ear tags and color coded cage cards.

Housing: Individually housed in wire mesh bottom cages.
Food: NIH Animal Feed A, certified feed, ad libitum, supplied fresh daily. No knowm 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

Type of coverage:

occlusive

Preparation of test site:

other: abraded and intact

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

A volume of 0.5 ml of the test article (undiluted) was applied to each of two sites per rabbit.

Duration of treatment / exposure:

4 hours

Observation period:

Four hours post-dose, the occlusive binders and patches were removed. The exposure sites were gently wiped with clean gauze to remove as much non-absorbed test article as possible. One-half hour after unwrapping, the exposure sites were examined and scored separately for both erythema and edema on a graded scale of 0 to 4 in accordance with the Skin Reaction Code (method of Draize, 1965). The exposure sites were again examined and scored at 48, 72 and 96 hours. Daily observations were not recorded, however, any extraordinary findings, dermal or toxicological, were noted.

Number of animals:

6 animals.

Details on study design:

Preparation:
On the day prior to dosing, the fur was clipped from the dorsal trunk area of each animal with electric clippers. During this procedure, care was taken to avoid accidental abrasion to the skin.
Method:
A volume of 0.5 ml was applied to each of two sites per rabbit. The skin sites were located directly across from each other, separated by the spinal column. Just prior to administration, test sites on the right dorsal side were abraded with the tip of a 22 gauge hypodermic needle. The abrasions were 4 incisions made in cross-hatch which penetrated the stratum corneum but did not disturb the dermis and/or produce bleeding. Following application, each skin site was occluded ,with a one-inch square gauze patch. The trunk of each animal was then wrapped with an occlusive binder which consisted of a layer of plastic wrap, a protective cloth and a stockinette sleeve all secured in place with masking tape.
Observation:
Four hours post-dose, the occlusive binders and patches were removed. The exposure sites were gently wiped with clean gauze to remove as much non-absorbed test article as possible. One-half hour after unwrapping, the exposure sites were examined and scored separately for both erythema and edema on a graded scale of 0 to 4 in accordance with the Skin Reaction Code (Draize) in Appendix II. The exposure sites were again examined and scored at 48, 72 and 96 hours. Daily observations were not recorded, however, any extraordinary findings, dermal or toxicological, were noted.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

other: Mean score for all animals

Time point:

other: overall at 24.5, 48 and 72 h

Score:

0

Max. score:

4

Remarks on result:

other: Intact skin

Results 2

Irritation parameter:

edema score

Basis:

other: Mean score for all animals

Time point:

other: overall at 24.5, 48 and 72 h

Score:

0

Max. score:

4

Remarks on result:

other: Intact skin

Any other information on results incl. tables

Individual Rabbit Skin Irritation Scores

			Rabbit Number
Parameter scored	Exposure Time (hours)	Skin	1123	1124	1125	1176	1177	1178
Erythema and Eschar Formation	24.5	Intact	0	0	0	0	0	0
	48	Intact	0	0	0	0	0	0
	72	Intact	0	0	0	0	0	0
	96	Intact	0	0	0	0	0	0
	24.5	Abraded	0	0	0	0	0	0
	48	Abraded	0	0	0	0	0	0
	72	Abraded	0	0	0	0	0	0
	96	Abraded	0	0	0	0	0	0
(A)Erythema and Eschar Sub-total			0	0	0	0	0	0
Edema Formation	24.5	Intact	0	0	0	0	0	0
	48	Intact	0	0	0	0	0	0
	72	Intact	0	0	0	0	0	0
	96	Intact	0	0	0	0	0	0
	24.5	Abraded	0	0	0	0	0	0
	48	Abraded	0	0	0	0	0	0
	72	Abraded	0	0	0	0	0	0
	96	Abraded	0	0	0	0	0	0
(B)Edema Sub-total			0	0	0	0	0	0
(C)Overall Total = (A+B)			0	0	0	0	0	0
(D) Individual Animal Scores = (C/8)			0	0	0	0	0	0

Mean Score (D/6) = 0

Initial Body Weights:

Animal number                     Initial Body Weight (kg)

82 -1123                                   2.00

82 -1124                                   2.46

82 -1125                                   2.35

82 -1176                                   2.30

82 -1177                                   2.92

82 -1178                                   2.57

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

There were no irritative effects observed in any of the animals on intact or abraded skin at any time point during the study. All animals appeared normal throughout the course of the study.

Executive summary:

The skin irritation potential of the test material was evaluated in this study with New Zealand White rabbits. One group of six albino rabbits received a volume of 0.5 ml of the test article applied to two sites per rabbit. One site consisted of intact skin and the other was abraded skin. Following the application, each site was occluded. Four hours post-dose, the occlusive binders were removed. The exposure sites were wiped to remove as much non-absorbed test article as possible. One-half hour after unwrapping, the exposure sites were examined and scored separately for both erythema and edema according to the method of Draize. The exposure sites were again examined and scored at 48, 72 and 96 hours. There were no irritative effects observed in any of the animals at any time-point. All animals appeard normal throughout the course of the study. The individual animal mean scores for the 24.5, 48 and 72 hr time-point for the intact skin indicated that this material would not be classified under EU regulations.