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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

disregarded due to major methodological deficiencies

Study period:

1969

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

This study is considered to be reliability 3 since it was performed at Industrial Biotest Laboratories prior to the implementation of GLP and during a time period where the conduct, quality and reliability of studies performed at this lab was brought into question. The study may have been performed in a satisfactory manner but insufficient information is available to allow a complete audit to verify the quality and reliability.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1969

Report date:

1969

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

No specific guideline referenced, however, methods of study were well documented (see details on dermal exposure and study design).

GLP compliance:

no

Remarks:

Study conducted prior to GLP's.

Test type:

other:

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 animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

Young adult, New Zealand strain albino rabbits ranging in body weight from 2.2 to 2.5 kilograms were employed as test animals. All rabbits had been maintained under observation in the laboratory for at least seven days prior to testing. During the pre-test period the animals were examined with respect to their general health and suitability as test animals. The rabbits were housed individually in hanging rabbit cages and maintained on a standard laboratory rabbit ration. Food and water were permitted ad libitum.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Twenty-four hours prior to the dermal applications, the backs of the rabbits were shaved free of hair with electric clippers. The shaved area on each animal constituted about 30 percent of the total body surface area. The animals were then returned to their cages to await testing on the following day. The 24-hour waiting period allowed recovery of the stratum corneum from the disturbance which accompanied the close-clipping procedure and also permitted healing of any microscopic abrasions possibly produced during the process.
On the testing day, the rabbits received skin applications of the undiluted test material at dose levels of 6.8 and 10.2 g/kg. A group of four rabbits (two male and two female) was tested at each level. After each application, the exposure site was covered by wrapping the trunk of the animal with an impervious plastic sheeting which was securely taped in place. This plastic wrap insured intimate contact of epidermis and test material. To further prevent oral ingestion of the test material, each animal was fitted with a light-weight flexible plastic collar which was worn through out the observation period.
The test material remained in contact with the skin for 24 hours. At the end of this period the plastic sheeting was taken off and all residual material removed. The exposure sites were examined for local skin reactions and the animals returned to their cages.

Duration of exposure:

24 hours

Doses:

6.8 and 10.2 g/kg

No. of animals per sex per dose:

Two male and two female per dose group.

Control animals:

no

Details on study design:

Observations for mortality, local skin reactions, and behavioral abnormalities were continued for a total of 14 days following the skin applications. Initial and final body weights were also recorded. Arrangements were made to autopsy any animals which might succumb during the study as well as all surviving animals at the end of the observation period.

Statistics:

At the end of the observation period, all data were collected and arrangements made to calculate, if possible, the acute dermal median lethal dose (LD50) of the test material using the techniques of Weil (1952), Thompson (1947), and Thompson and Weil (1952).

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 10.2 other: g/kg

Based on:

test mat.

Mortality:

There were no deaths.

Clinical signs:

other: No untoward behavioral reactions were noted among any of the animals. Wrinkling of the skin at the application site was noted among animals in both groups by the seventh day of the 14-day observation period. No improvement was noted by the end of the 14-d

Gross pathology:

No gross pathologic alterations were noted among any of the animals other than the dermal alterations previously described.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

There was no mortality observed in male and female rabbits dosed with 6.8 and 10.2 g/kg. The acute dermal LD50 of Automate Blue No. 8 was determined to be > 10.2 g/kg body weight in male and female NZW rabbits.

Executive summary:

An acute dermal toxicity study was conducted using male and female New Zealand White rabbits given a single dermal application of a dose of 6.8 or 10.2 g/kg body weight Automate Blue No.8 (2 males and 2 female per dose group). No mortality was observed in either dose group. One animal in the 6.8 g/kg dose group and three animals in the 10.2 g/kg dose group lost weight. No untoward behavioral reactions were noted among any of the animals. Wrinkling of the skin at the application site was noted among animals in both groups by the seventh day of the 14-day observation period. No improvement was noted by the end of the 14-day observation period. The dark blue color of the test material precluded evaluation of the skin at the application site for erythema. No gross pathologic alterations were noted among any of the animals other than the dermal alterations previously described. The acute dermal LD50 of Automate Blue No. 8 was determined to be > 10.2 g/kg body weight in male and female NZW rabbits.