Pyrrolo[3,4-c]pyrrole-1,4-dione, 3,6-bis([1,1'-biphenyl]-4-yl)-2,5-dihydro-

Endpoint summary

Description of key information

- Skin: according to OECD TG 404, GLP, rabbit, not irritating
- Eye: according to OECD TG 405, GLP, rabbit, not irritating

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

Study period:

1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

Erythema could not be scored due to staining of the skin by the test substance.

GLP compliance:

yes

Specific details on test material used for the study:

- Substance type: Organic
- Physical state: Solid
- Lot/batch No.: Z-2281/1,2,4,5
- Expiration date of the lot/batch: April 01, 1997
- Storage condition of test material: at room temperature in the dark

Species:

rabbit

Strain:

New Zealand White

Remarks:

Albino, SPF-Quality

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: Approx. 12 weeks
- Weight at study initiation: 2287 - 2600 grams
- Housing: individual
- Diet: Standard laboratory rabbit diet, approx. 100 g per day
- Water: Tap-water diluted with decalcified water, ad libitum
- Acclimation period: At least 5 days before start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 55
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1992-11-24 To: 1992-11-27

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g
- The powdery test substance was applied with the aid of a patch. The patch was moistened with distilled water, immediately before application, to ensure close contact of the test substance to the animal's skin.

Duration of treatment / exposure:

4 h

Observation period:

72 h (Observations were made 1, 24, 48 and 72 hours after exposure)

Number of animals:

3 male rabbits

Details on study design:

TEST SITE
- Approximately 24 hours before treatment, the dorsal fur was shaved with electric clippers
- Area of exposure: approximately 150 square centimeters (10 cm X 15 cm).
- Type of wrap if used: moistened Scotchpak-non-woven patch, mounted on Micropore tape. The dressing was wrapped around the abdomen and secured with an elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the remaining test substance was removed using a tissue moistened with tap-water and subsequently a dry tissue
- Time after start of exposure: 4h

SCORING SYSTEM: Draize
- ERYTHEMA AND ESCHAR FORMATION
0 No erythema
1 Very slight erythema (barely perceptible)
2 Well defined erythema
3 Moderate to severe erythema
4 Severe erythema (beet redness) to slight eschar formation (injuries in depth)

- OEDEMA FORMATION
0 No oedema
1 Very slight oedema (barely perceptible)
2 Slight oedema (edges of area well defined by definite raising)
3 Moderate oedema (raised approximately 1 mm)
4 Severe oedema (raised more than 1 mm and extending beyond area of exposure)

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

- Erythema: Erythema could not be scored due to staining of the skin by the test substance.
- Microscopic findings: No abnormalities were detected in either the treated skin-sites and reference control skin-sites by microscopical examination. The absence of microscopical detectable lesions in the skin, 72 hours after exposure, indicate that no or negligible irritation has occurred during the study period.
- Corrosion: There was no evidence of a corrosive effect on the skin.

Other effects:

Toxicity symptoms / mortality: No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1: Individual findings

Observation time	Rabbit no. 4266		Rabbit no. 4268		Rabbit no. 4270
	Erythema	Oedema	Erythema	Oedema	Erythema	Oedema
1 Hour	-	0	-	0	-	0
24 Hours	-	0	-	0	-	0
48 Hours	-	0	-	0	-	0
72 Hours	-	0	-	0	-	0
Mean Value	-	0.0	-	0.0	-	0.0

Interpretation of results:

GHS criteria not met

Conclusions:

The facts that no oedema was observed during the study and microscopical examination of the treated skin sites after 72 hours revealed no abnormalities indicate that the test substance is not irritanting to skin.

Executive summary:

A GLP conform in vivo study was performed to assess the acute dermal irritation/corrosion potential of the test substance (purity 97.5%) in White New Zealand rabbits according to the OECD guideline 404. Three male rabbits were exposed to 0.5 g of the test substance that was applied onto shaved skin using a semi-occlusive dressing. After 4 hours, the remaining test substance was removed. Observations were made 1, 24, 48 and 72 hours after exposure. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. No oedema in the treated skin-areas of the rabbits were observed. Erythema could not be scored due to red staining of the skin by the test substance. However, microscopical examination revealed no abnormalities in either the treated skin-sites and reference control skin-sites 72 hours after exposure. The absence of microscopical detectable lesions in the skin indicate that no or negligible irritation has occurred during the study period. Therefore, the test substance is considered as non-irritating when applied to intact rabbit skin.

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

Study period:

1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

- Substance type: Organic
- Physical state: Solid
- Lot/batch No.: Z-2281/1,2,4,5
- Expiration date of the lot/batch: April 01, 1997
- Storage condition of test material: at room temperature in the dark

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: Approx. 15 weeks
- Weight at study initiation: 2666 - 3022 grams
- Housing: individual
- Diet: Standard laboratory rabbit diet, approx. 100 g per day
- Water: Tap-water diluted with decalcified water, ad libitum
- Acclimation period: At least 5 days before start of treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C
- Humidity (%): 55
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1992-11-30 To: 1992-12-03

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

21 mg

Duration of treatment / exposure:

single

Observation period (in vivo):

3 days

Number of animals or in vitro replicates:

3 male rabbits

Details on study design:

REMOVAL OF TEST SUBSTANCE: No

SCORING SYSTEM: Draize

- Cornea opacity degree of density (area most dense taken for reading)
0 No ulceration or opacity
1 Scattered or diffuse areas of opacity (other than slight dulling of normal lustre), details of iris clearly visible
2 Easily discernible translucent area, details of iris slightly obscured
3 Nacreous areas, no details of iris visible, size of pupil barely discernible
4 Opaque cornea, iris not discernible through the opacity

- Area of cornea affected:
0 No ulceration or opacity
1 One quarter or less but not zero
2 Greater than one quarter, but less than half
3 Greater than half, but less than three quarters
4 Greater than three quarters, up to whole area

- Iris:
0 Normal
1 Markedly deepened rugae, congestion, swelling, moderate circum corneal hyperaemia, or injection, any of these or any combination thereof, iris still reacting to light (sluggish reaction is positive)
2 No reaction to light, haemorrhage, gross destruction (any or al 1 of these)

- Conjuctivae redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris):
0 Blood vessels normal
1 Some blood vessels definitely hyperaemic (injected)
2 Diffuse, crimson colour, individual vessels not easily discernible
3 Diffuse beefy red

- Conjuctivae chemosis: lids and/or nictitating membrane
0 No swelling
1 Any swelling above normal (includes nictitating membrane)
2 Obvious swelling with partial eversion of lids
3 Swelling with lids about half closed
4 Swelling with lids more than half closed

- Conjuctivae discharge:
0 No discharge 0
1 Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2 Discharge with moistening of the lids and hairs, just adjacent to lilds
3 Discharge with moistening of the lids and hairs, in a considerable area around the eye

TOOL USED TO ASSESS SCORE: fluorescein
Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

24/48/72 h

Score:

0.3

Max. score:

1

Reversibility:

fully reversible within: 48h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

- Irritation: Instillation of the test substance into one eye of each of the three male albino rabbits affected the conjunctivae in one animal. The irritation of the conjunctivae had resolved within 48 hours after instillation. A small amount of discharge was observed at the 24 hour observation in another animal. Eye irritation was not observed at all in the third animal during the study period. Treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals.
- Corrosion: There was no evidence of ocular corrosion.

Other effects:

- Colouration: Red/purple staining of the fur on the head, surrounding the treated eye, by the test substance was observed.
- Toxic symptoms / mortality: No toxic symptoms were observed in the animals during the test period and no mortality occurred.

Table 1: Individual Draize score calculation

Animal No.	Tissue	Hours after administration
		1	24	48	72
4218	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae	2	4	0	0
	Subtotal	2	4	0	0
4227	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae	0	2	0	0
	Subtotal	0	2	0	0
4299	Cornea	0	0	0	0
	Iris	0	0	0	0
	Conjunctivae	0	0	0	0
	Subtotal	0	0	0	0
Total		2	6	0	0
Mean total		0.7	2	0	0

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study, the test substance is considered not irritanting to eyes.

Executive summary:

To assess the acute eye irritation/corrosion potential of the test substance (purity 97.5%), a GLP conform eye irritation test in White New Zealand rabbits was performed according to OECD guideline 405. Single samples of approximately 21 mg of the test substance were instilled into one eye of each of three male rabbits. Observations were made 1, 24, 48 and 72 hours after instillation. No signs of systemic intoxication were observed and no mortality occurred during the study period. Instillation of the test substance resulted in effects on the conjunctivae in two animals. Irritation of the conjunctivae was observed as slight redness and discharge in one animal each, which had completely resolved within 48 hours. No corneal opacity was observed over the entire study period and treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals. Mean scores of the 24-72 h readings were 0.0 for both corneal opacity and iris, 0.33 for conjunctivae redness. Therefore, the test substance is considered non-irritating to the rabbit eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin

A GLP conform in vivo study was performed to assess the acute dermal irritation/corrosion potential of the test substance (purity 97.5%) in White New Zealand rabbits according to the OECD guideline 404. Three male rabbits were exposed to 0.5 g of the test substance that was applied onto shaved skin using a semi-occlusive dressing. After 4 hours, the remaining test substance was removed. Observations were made 1, 24, 48 and 72 hours after exposure. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. No oedema in the treated skin-areas of the rabbits were observed. Erythema could not be scored due to red staining of the skin by the test substance. However, microscopical examination revealed no abnormalities in either the treated skin-sites and reference control skin-sites 72 hours after exposure. The absence of microscopical detectable lesions in the skin indicate that no or negligible irritation has occurred during the study period. Therefore, the test substance is considered as non-irritating when applied to intact rabbit skin.

 

Eye

To assess the acute eye irritation/corrosion potential of the test substance (purity 97.5%), a GLP conform eye irritation test in White New Zealand rabbits was performed according to OECD guideline 405. Single samples of approximately 21 mg of the test substance were instilled into one eye of each of three male rabbits. Observations were made 1, 24, 48 and 72 hours after instillation. No signs of systemic intoxication were observed and no mortality occurred during the study period. Instillation of the test substance resulted in effects on the conjunctivae in two animals. Irritation of the conjunctivae was observed as slight redness and discharge in one animal each, which had completely resolved within 48 hours. No corneal opacity was observed over the entire study period and treatment of the eyes with 2% fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage in any of the animals. Mean scores of the 24-72 h readings were 0.0 for both corneal opacity and iris, 0.33 for conjunctivae redness. Therefore, the test substance is considered non-irritating to the rabbit eye.

 

No data is available on respiratory irritation.

Justification for classification or non-classification

Classification, Labeling, 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 as amended for the thirteenth time in Regulation (EC) No. 2018/1480.