N,N'-[6,13-diacetamido-2,9-diethoxy-3,10-triphenodioxazinediyl]bis(benzamide)

Endpoint summary

Description of key information

The substance has a primary irritation index of 0.4 and is therefore a minimal primary skin irritant in the albino rabbit. In a second skin irritation study, the substance did not exhibit skin irritating properties.
The substance did not show eye irritating properties. In a second eye irritation study, the substance was determined to be a minimal eye irritant.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Qualifier:

according to guideline

Guideline:

other: "Hazardous Substances Regulations" under the U.S. Federal Hazardous Substances Labelling Act Sect. 191.11 (February 1965)

GLP compliance:

no

Remarks:

pre-GLP

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: bred on the premises
- Age at study initiation: 3 - 4 months
- Weight at study initiation: male 2.20 kg and female 2.32 kg
- Housing: single
- Diet: commercial irradiated diet (Styles-Oxoid) was fed ad libitum
- Water: sterile filtered water was available at all times
- Acclimation period: one week prior to the trial

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±1
- Humidity (%): 50 - 70
- Photoperiod (hrs dark / hrs light): artificial light for 10 hours daily from 8:00 - 18:00 hours

Type of coverage:

occlusive

Preparation of test site:

other: left shaved; right shaved+abraded

Vehicle:

other: 50% solution of polyethylene glycol and water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5g

VEHICLE
- Amount(s) applied (volume or weight with unit): 15 mL
- Concentration (if solution): 50% solution of polyethylene glycol and water

Duration of treatment / exposure:

24 hours

Observation period:

72 hours

Number of animals:

6 (3 males, 3 females)

Details on study design:

TEST SITE
- Area of exposure: 2 test sites of 2.5 cm2
- % coverage: 10%
- Type of wrap if used: Gauze pad covered with aluminium foil secured with "Sleek" adhesive tape. The test sites were then enclosed by a 6" wide "Coban"-self adhesive bandage the edges of which were fixed to the skin by strips of "Sleek" in order to retain the test substance in close contact with the skin.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No
- Time after start of exposure: 24 hours

SCORING SYSTEM: Draize

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: intact and abraded skin

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 hours

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: intact skin

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 hours

Score:

0.83

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: abraded skin

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: intact and abraded skin

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24 hours

Score:

0.4

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: intact and abraded skin

Irritant / corrosive response data:

A very slight to slight oedema was present in 5/6 rabbits 24 hours after application of the compound. All sites were clear by 72 hours.
There was no significant difference between intact and abraded sites.

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Remarks:

After application eye lids were held open for a few seconds instead of closed.

Qualifier:

according to guideline

Guideline:

other: "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965)

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: bred on the premises
- Age at study initiation: 10-12 weeks
- Weight at study initiation: male 2.20 kg and female 2.32 kg
- Housing: single
- Diet: A commercial irradiated diet (Styles-Oxoid) was fed ad libitum
- Water: Sterile filtered water was available at all times
- Acclimation period: one week prior to the start of the trial

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20±1
- Humidity (%): 50-70
- Photoperiod (hrs dark / hrs light): artificial light for 10 hours daily from 08.00 - 18.00 hours

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

Eyes of 3 animals were rinsed 30 seconds after treatment. Eyes of 3 animals were not rinsed.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

6

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the eyes of three animals, with warm water
- Time after start of exposure: 30 seconds

SCORING SYSTEM: Draize

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 1 hour

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eyes not washed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 6 hours

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eyes not washed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 1 day

Score:

0.6

Max. score:

3

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eyes not washed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 2 days

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 3 days

Remarks on result:

other: eyes not washed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 1 hour

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 1 day

Remarks on result:

other: eyes washed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 6 hours

Score:

0.6

Max. score:

3

Reversibility:

fully reversible within: 1 days

Remarks on result:

other: eyes washed

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 1 days

Remarks on result:

other: eyes not washed

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 6 hours

Score:

0.6

Max. score:

4

Reversibility:

fully reversible within: 1 day

Remarks on result:

other: eyes not washed

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 1 hour

Score:

0.6

Max. score:

4

Reversibility:

fully reversible within: 6 hours

Remarks on result:

other: eyes washed

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

mean

Time point:

other: 1 hour

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 1 day

Remarks on result:

other: eyes not washed

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

mean

Time point:

other: 6 hours

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 1 day

Remarks on result:

other: eyes not washed

Irritation parameter:

conjunctivae score

Remarks:

discharge

Basis:

mean

Time point:

other: 1 hour

Score:

0.6

Max. score:

3

Reversibility:

fully reversible within: 6 hours

Remarks on result:

other: eyes washed

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance is a minimal eye irritant in the albino rabbit.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In a skin irritation test 6 New Zealand White rabbits (3 male, 3 female) were exposed to 0.5 g of the test substance in a 50% solution of polyethylene glycol and water. Twenty-four hours prior to the dermal application, the backs of the rabbits were shaved over an area consisting of at least 10% of the total body surface. Two test sites lateral to the mid line of the back were used on each rabbit. Immediately before the application of the test compound, the right hand site was abraded. The application site was covered with aluminium foil secured with "Sleek" adhesive tape. The test sites were then enclosed by a 6" wide "Coban"- self adhesive bandage the edges of which were fixed to the skin by strips of "Sleek". After 24 hours the dressing was removed and the intact and abraded application sites were assessed for oedema and erythema. A further assessment was made at 72 hours. A very slight to slight oedema was present in 5/6 rabbits 24 hours after application of the compound. All sites were clear by 72 hours. There was no significant difference between intact and abraded sites. The primary irritation score was 0.4. The substance is a minimal primary skin irritant in the albino rabbit.

 

In a skin irritation test 6 Russian breed rabbits (3 male, 3 female) were exposed to 0.5 g of the test substance wetted with tap water. One day before treatment, the flanks of the rabbits were shaved with an electric clipper. The shaved skin area on the left side was slightly scarified just before treatment. A gauze patch of 2.5 x 2.5 cm laden with the test substance was applied immediately to the prepared skin. The patch was covered with an impermeable foil of 5 x 5 cm, which was fixed to the body of the animals with adhesive tape. The gauze patches were removed 24 hours after application. The reaction of the skin was appraised upon removal during an observation period of 7 days. No signs of erythema or oedema were observed up to 7 days after application of the compound. Therefore, under the conditions of this experiment the substance was found to be non irritant when applied to intact and abraded rabbit skin.

 

In a eye irritation test 6 Russian breed rabbits (3 male, 3 female) received 0.1 g of the test substance into the conjunctival sac of the left eye with a spatula. After application, the eyelids were held open for a few seconds. The right eye served as a control. The treated eyes of three rabbits were each rinsed with physiologic saline about 1 minute after treatment. The reactions were appraised with a slit-lamp after 1 and 6 hours and after 1, 2, 3, 6 and 8 days. The irritation index was found to be 0 for the cornea, 0 for the iris and 0 for the conjunctivae. Therefore the test substance is to be considered as non-irritant to the eye of rabbits.

 

In a eye irritation test 6 Russian breed rabbits (3 male, 3 female) received 100 mg of the test substance into the conjunctival sac of the left eye with a spatula. After application, the eyelids were held closed for one second. The right eye served as a control. After 30 seconds the treated eyes of three rabbits were each rinsed with warm water for 1 minute after treatment. The rabbits were examined 1, 6, 24, 48 and 72 hours after application. A minimal conjunctival reaction was seen one hour after the application of the compound. Only 2/3 unwashed eyes were still showing a minimal reaction at 24 hours and all were clear by 72 hours.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for skin and eye irritation under Regulation (EC) No. 1272/2008.