Reaction product of 3,9-dibromobenzanthrone condensed with 2 equivalents of 1-aminoanthraquinone, subsequently further condensed under oxidative conditions

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

In vivo study in rats. At the end of the observation period, the acute oral median lethal dose (LD50) of the test material was calculated.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Young albino rats derived from Sprague-Dawley stock were used as test animals. All animals were kept under observation for five days prior to experimental use, during which period they were checked for general health and suitability as test animals. The animals were housed in stock cages and were permitted a standard laboratory diet plus water ad libitum, except during the 16-hour period immediately prior to oral intubation when food was withheld.

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

No data

Doses:

1350, 4556, 15380 mg/kg

No. of animals per sex per dose:

4 animals per group (2 male/2 female) - total of 12 animals

Control animals:

no

Details on study design:

Initial screening was conducted in order to determine the general level of toxicity of the test material. Selected groups of albino rats were administered the test material at several dose levels. All doses were administered directly into the stomachs of the rats using a hypodermic syringe equipped with a ball-tipped intubating needle.After oral administration of the test material, the rats were housed individually in suspended, wire-mesh cages and observed for the following 14 days. Initial and final body weights, mortalities, and reactions were recorded. A necropsy examination was conducted on all animals.

Statistics:

At the end of the observation period, the acute oral median lethal dose (LD50) of the test material was calculated, if possible, using the techniques of Weil, Thompson, and Thompson and Weil.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 15 380 mg/kg bw

Based on:

test mat.

Mortality:

No mortality observed

Clinical signs:

other: No pharmacotoxic symptoms were exhibited by the rats dosed at 1,350 and 4,556 mg/kg following oral administration of CIBANONE Olive S Paste. In rats dosed at 15,380 mg/kg hypoactivity was observed at 1 hour, ruffed fur at 3 hours, and black faeces at 5 ho

Gross pathology:

Necropsy examination did not reveal any gross pathologic alterations.

Other findings:

Areas on the fur and tail remained stained throughout the 14-day observation period.

Mortality and Body Weight Data

Dose Level (mg/kg)	Animal Number and Sex	Individual Body Weight (grams)		Number dead	Percent dead
		Test Day Number:
		0	14	Number Tested
1,350	1-M 2-M 3-F 4-F	214 222 170 186	300 320 220 234	0/4	0
4,556	5-M 6-M 7-F 8-F	210 210 170 170	310 320 236 222	0/4	0
15,380	9-M 10-M 11-F 12-F	240 202 164 180	340 310 230 248	0/4	0

 

Interpretation of results:

GHS criteria not met

Conclusions:

Acute Oral LD50: >15,380 mg/kg

Executive summary:

Young albino rats derived from Sprague-Dawley stock were used as test animals.

Selected groups of albino rats were administered the test material at several dose levels. All doses were administered directly into the stomachs of the rats using a hypodermic syringe equipped with a ball-tipped intubating needle.

After oral administration of the test material, the rats were housed individually in suspended, wire-mesh cages and observed for the following 14 days. Initial and final body weights, mortalities, and reactions were recorded. A necropsy examination was conducted on all animals.

At the end of the observation period, the acute oral median lethal dose (LD50) of the test material was calculated.

 

No pharmacotoxic symptoms were exhibited by the rats dosed at 1,350 and 4,556 mg/kg following oral administration of CIBANONE Olive S Paste. In rats dosed at 15,380 mg/kg hypoactivity was observed at 1 hour, ruffed fur at 3 hours, and black feces at 5 hours. These symptoms subsided after 3 days. However areas on the fur and tail remained stained throughout the 14-day observation period.

Necropsy examination did not reveal any gross pathologic alterations.

 

Acute Oral LD50: >15,380 mg/kg

Cross-reference 2

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The test procedure was modelled after that of Draize et al.

GLP compliance:

no

Specific details on test material used for the study:

Black liquid identified as CIBANONE Olive S Paste, 44/121695/100/0

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Young albino rabbits of the New Zealand strain were used in the evaluation of the primary skin irritating properties of the test material.

Type of coverage:

occlusive

Preparation of test site:

other: Shaved & abraded

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

0.5ml, undiluted

Duration of treatment / exposure:

24 hours exposure period

Observation period:

72 hours

Number of animals:

6 animals (sex not disclosed)

Details on study design:

Prior to the application of the test material, the hair was clipped from the back and flanks of each rabbit. Two test sites located lateral to the midline of the back approximately ten centimeters apart were selected. One of the two sites was abraded by making four epidermal incisions, two perpendicular to the other two, while the other test site remained intact.The test material was applied to each of the test sites on each rabbit and occluded with gauze patches which were secured with masking tape. The trunk of each animal was then wrapped with impervious plastic sheeting. The wrap held the patches in position and retarded evaporation of the test material during the 24-hour exposure period.At the end of 24 hours, the plastic wrappings, patches and all residual test material were removed. The intact and abraded test sites were examined and scored separately for erythema and edema on a graded scale of 0 to 4. After 72 hours, the sites were again examined and scored.In evaluating the average irritation present, the mean scores for erythema and edema of the intact test sites after 24 and 72 hours were added. Similarly, the mean scores for erythema and edema of the abraded test sites after 24 and 72 hours were added. These two values were totalled and divided by four to obtain the mean primary irritation score.

Irritation parameter:

erythema score

Remarks:

Intact Skin Sites

Basis:

mean

Time point:

24 h

Score:

2.7

Max. score:

4

Reversibility:

not fully reversible within: 72 h

Irritation parameter:

erythema score

Remarks:

Intact skin

Basis:

mean

Time point:

72 h

Score:

2.8

Max. score:

4

Reversibility:

not fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

mean

Time point:

24 h

Score:

1.7

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Irritation parameter:

edema score

Remarks:

Intact skin

Basis:

mean

Time point:

72 h

Score:

0.3

Max. score:

4

Reversibility:

not fully reversible within: 72 h

Irritation parameter:

erythema score

Remarks:

Abraded skin

Basis:

mean

Time point:

24 h

Score:

2.7

Max. score:

4

Reversibility:

not fully reversible within: 72 h

Irritation parameter:

erythema score

Remarks:

Abraded skin

Basis:

mean

Time point:

72 h

Score:

3

Max. score:

4

Reversibility:

not fully reversible within: 72 h

Irritation parameter:

edema score

Remarks:

Abraded skin

Basis:

mean

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 72 hours

Irritation parameter:

edema score

Remarks:

Abraded skin

Basis:

mean

Time point:

72 h

Score:

0.5

Max. score:

4

Reversibility:

not fully reversible within: 72 h

Irritant / corrosive response data:

Extremely superficial second degree chemical burns were noted at the 24-hour scoring interval on all sites. The burns were considered too superficial to be given a maximum score of four for both erythema and edema.

Other effects:

None reported

Primary Skin Irritation - Results

Animal Number	Irritation Scores for Abraded Skin Sites at:				Irritation Scores for Intact Skin Sites at:
	24 Hours		72 Hours		24 Hours		72 Hours
	Er.	Ed.	Er.	Ed.	Er.	Ed.	Er.	Ed.
1 2 3 4 5 6	2 2 3 3 3 3	2 2 2 2 2 2	3 3 3 3 3 3	1 1 0 0 0 1	2 2 3 3 3 3	1 2 1 2 2 2	2 3 3 3 3 3	0 1 0 1 0 0
Mean	2.7	2.0	3.0	0.5	2.7	1.7	2.8	0.3
Subtotal	8.2				7.5
Primary Irritation Score = 3.9

Er. = Erythema

Ed. = Edema

Interpretation of results:

other: Moderately irritating

Conclusions:

Mean Primary Irritation Score: 3.9/8.0. The test material was moderately irritating to rabbit skin.

Executive summary:

Young albino rabbits of the New Zealand strain were used in the evaluation of the primary skin irritating properties of the test material.

The test procedure was modelled after that of Draize et al.

 

Prior to the application of the test material, the hair was clipped from the back and flanks of each rabbit. One of the two sites was abraded by making four epidermal incisions, two perpendicular to the other two, while the other test site remained intact.

The test material was applied to each of the test sites on each rabbit and occluded with gauze patches which were secured with masking tape. The wrap held the patches in position and retarded evaporation of the test material during the 24-hour exposure period.

At the end of 24 hours, the plastic wrappings, patches and all residual test material were removed. The intact and abraded test sites were examined and scored separately for erythema and edema on a graded scale of 0 to 4. After 72 hours, the sites were again examined and scored.

 

Extremely superficial second degree chemical burns were noted at the 24-hour scoring interval on all sites. The burns were considered too superficial to be given a maximum score of four for both erythema and edema.

 

Mean Primary Irritation Score: 3.9/8.0

Moderately Irritating

Cross-reference 3

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The test method was patterned after that of Draize et al.

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Young albino rabbits of the New Zealand strain were used to evaluate the eye irritating properties of the test material.

Vehicle:

unchanged (no vehicle)

Controls:

other: Untreated eyes acted as controls

Amount / concentration applied:

0.1ml undiluted

Duration of treatment / exposure:

Single exposure

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6 animals (sex not disclosed)

Details on study design:

The test material was instilled into the conjunctival sac of the right eye of each rabbit. The left eye of each animal served as a control. At each scoring interval the cornea, iris, and palpebral conjunctiva were examined and graded for irritation and injury according to a standard scoring system. The maximum possible score at any one examination and scoring period is 110 points, which indicates maximal irritation and damage to all three ocular tissues. Zero score indicates no irritation. In this scoring system, special emphasis is placed upon irritation or damage to the cornea, while less emphasis is placed upon damage to the iris and conjunctiva.After the completion of the test, the scores were analyzed, and a descriptive eye irritation rating was assigned to the test material. The criteria used for assignment of the descriptive rating are the frequency, the extent, and the persistence of irritation or damage which occur to the three ocular tissues.The rating is obtained by selecting the maximum mean irritation score at one, 24, 48 or 72 hours after instillation. If the rate of dissipation of injury does not meet the requirements defined for the descriptive rating appropriate for a particular numerical score, the descriptive rating is raised by one or more levels.

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #4

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #5

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

cornea opacity score

Basis:

animal #6

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #4

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #5

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

animal #6

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #4

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #5

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

animal #6

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: not applicable

Irritant / corrosive response data:

All animals had a score of 2.0 for conjunctiva at the 1 hour time point. In all animals this score had reduced to score by 24 hours.

Other effects:

None reported

Eye Irritation Test - Results

Tissue	Rabbit Number	1 Hour	24 Hours	48 Hours	72 Hours	7 Days
Cornea (D-A) Iris Conjunctivae (R-S-D)	1	0 0 2 (1-0-0)	0 0 0	0 0 0	0 0 0	0 0 0
Total		2	0	0	0	0
Cornea (D-A) Iris Conjunctiva (R-S-D)	2	0 0 2 (1-0-0)	0 0 0	0 0 0	0 0 0	0 0 0
Total		2	0	0	0	0
Cornea (D-A) Iris Conjunctivae (R-S-D)	3	0 0 2 (1-0-0)	0 0 0	0 0 0	0 0 0	0 0 0
Total		2	0	0	0	0
Cornea (D-A) Iris Conjunctivae (R-S-D)	4	0 0 2 (1-0-0)	0 0 0	0 0 0	0 0 0	0 0 0
Total		2	0	0	0	0
Cornea (D-A) Iris Conjunctivae (R-S-D)	5	0 0 2	0 0 0	0 0 0	0 0 0	0 0 0
Total		2	0	0	0
Cornea (D-A) Iris Conjunctivae (R-S-D)	6	0 0 2 (1-0-0)	0 0 0	0 0 0	0 0 0	0 0 0
Total		2	0	0	0	0
Average Cornea Iris Conjunctivae		0.0 0.0 2.0	0.0 0.0 0.0	0.0 0.0 0.0	0.0 0.0 0.0	0.0 0.0 0.0
Total		2.0	0.0	0.0	0.0	0.0

Cornea:                                Iris:                              Conjunctivae:

D = Density                          Iris Score = Value x 5          R = Redness

A = Are                               Maximum Score = 10          S = Swelling

Corneal Score = D x A x 5                                              D = Discharge

Maximum Score = 80                                                   Conjunctivae Score = (R+S+D) x 2

                                                                             Maximum Score = 20

Interpretation of results:

GHS criteria not met

Conclusions:

Irritation score (Mean/Maximum): 2.0/110.0
Practically Non-irritating

Executive summary:

Young albino rabbits of the New Zealand strain were used to evaluate the eye irritating properties of the test material. The test method was patterned after that of Draize et al.

The test material was instilled into the conjunctival sac of the right eye of each rabbit. The left eye of each animal served as a control. At each scoring interval the cornea, iris, and palpebral conjunctiva were examined and graded for irritation and injury according to a standard scoring system.

 

Irritation score (Mean/Maximum): 2.0/110.0

Practically Non-irritating

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1975

Report date:

1975

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Acute dermal toxicity study in rabbits. Observations were conducted for 14 days following dermal application.

GLP compliance:

no

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction product of 3,9-dibromobenzanthrone condensed with 2 equivalents of 1-aminoanthraquinone, subsequently further condensed under oxidative conditions

EC Number:

944-232-9

Molecular formula:

Not available - UVCB substance

IUPAC Name:

Reaction product of 3,9-dibromobenzanthrone condensed with 2 equivalents of 1-aminoanthraquinone, subsequently further condensed under oxidative conditions

Test material form:

liquid

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

Young adult albino rabbits of the New Zealand strain were used 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 suspended, wire-bottomed cages and maintained on a standard laboratory ration. Food and water were offered 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 permitted healing of any microscopic abrasions possibly produced during the process.The test material was applied at the highest reasonable dose level. The test site was covered by wrapping the trunk of the animal with impervious plastic sheeting which was securely taped in place. This plastic wrap insured close contact of the epidermis and test material. To prevent oral ingestion of the test material, each animal was fitted with a light-weight, flexible plastic collar which was worn throughout the observation period.At the end of the test period, the plastic sheeting and all residual test material were removed.

Duration of exposure:

24 hours

Doses:

3000 mg/kg

No. of animals per sex per dose:

4 animals (2 male/2 female)

Control animals:

no

Details on study design:

The test sites were examined for local skin reactions and the animals were returned to their cages. Observations for mortality, local skin reactions, and behavioral abnormalities were continued for a total of 14 days following the skin applications. Initial, 7 and 14-day body weights were recorded. A necropsy examination was conducted on all animals.In the case of significant mortality following the initial study, additional experiments were conducted at lower dose levels in order to obtain data sufficient to determine the acute dermal median lethal dose (LD50).

Statistics:

At the end of the observation period, the acute dermal median lethal dose (LD50) of the test material was calculated, if possible, using the techniques of Weil, Thompson, and Thompson and Weil.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 3 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality reported

Clinical signs:

other: No pharmacotoxic symptoms were exhibited by any of the rabbits following dermal exposure to CIBANONE Olive S Paste.The test material was moderately to severely irritating to the skin of the albino rabbit. Skin changes at 24 hours were characterized by red

Gross pathology:

Necropsy examination revealed red lungs in rabbit No. 1-M.No other gross pathologic alterations were noted except for the local skin changes as previously described.

Other findings:

None reported

Any other information on results incl. tables

Mortality and Body Weight Data

Dose Level (mg/kg)	Animal Number and Sex	Individual Body Weights (kg)			Number Dead	Percent Dead
		Test Day Number:
		0	7	14	Number Tested
3,000	1-M* 2-M 3-F* 4-F	2.78 2.66 2.90 2.92	2.86 2.72 2.96 3.00	2.90 2.82 3.12 3.10	0/4	0

* The skin at the site of application was abraded

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Acute Dermal LD50: > 3,000 mg/kg

Executive summary:

Young adult albino rabbits of the New Zealand strain were used as test animals.

 

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 test material was applied at the highest reasonable dose level. The test site was covered by wrapping the trunk of the animal with impervious plastic sheeting which was securely taped in place. This plastic wrap insured close contact of the epidermis and test material.

 

The test material remained in contact with the skin for 24 hours. At the end of this period, the plastic sheeting and all residual test material were removed. The test sites were examined for local skin reactions and the animals were returned to their cages. Observations for mortality, local skin reactions, and behavioral abnormalities were continued for a total of 14 days following the skin applications. Initial, 7 and 14-day body weights were recorded. A necropsy examination was conducted on all animals.

 

No pharmacotoxic symptoms were exhibited by any of the rabbits following dermal exposure to CIBANONE Olive S Paste.

The test material was moderately to severely irritating to the skin of the albino rabbit. Skin changes at 24 hours were characterized by red, well-defined erythema, moderate edema and superficial second degree burns. Mild desquamation was observed at 7 and 14 days.

Necropsy examination revealed red lungs in rabbit No. 1-M.

No other gross pathologic alterations were noted except for the local skin changes as previously described.

 

Acute Dermal LD50: > 3,000 mg/kg