2-cyano-2-[2,3-dihydro-3-(tetrahydro-2,4,6-trioxo-5(2H)-pyrimidinylidene)-1H-isoindol-1-ylidene]-N-methylacetamide

Endpoint summary

Description of key information

Skin irritation:

Key: in vivo, rabbit: not irritanting (non-GLP, similar to OECD 404, 1981)

Eye irritation:

Key: in vivo, rabbit: not irritanting (non-GLP, equivalent to OECD 405, 1981)

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:

comparable to guideline study with acceptable restrictions

Remarks:

acceptable restrictions: occlusive conditions, 72 h reading missing, 8-day observation period

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

occlusive conditions, 72 h reading missing, limited documentation

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Gaukler Offenbach/Main, Germany
- Weight at study initiation: male 2.98 kg, female 3.72 kg
- Housing: one animal per cage of stainless steel with wire mesh walk floors, floor area: 40 cm x 51 cm,
- Diet (e.g. ad libitum): Ovator Solikanin 4 mm; Muskator-Werke, Duesseldorf , Germany (about 130 g per animal per day)
- Water (e.g. ad libitum): About 250 ml per animal per day: demineralized water (Monday-Friday), tap water (Saturday/Sunday)
- Acclimation period: About 8 days before the beginning of the study in the skin laboratory; same housing conditions as during the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 26
- Humidity (%): Conventional (about 30 - 70%)
- Photoperiod (hrs dark / hrs light): 12 h/12 h

Type of coverage:

occlusive

Preparation of test site:

other: clipping of the fur at least 15 hours before beginning of the study

Vehicle:

water

Controls:

other: Untreated skin sites of the same animal

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g of the test substance is applied
- Concentration (if solution): 50 % aqueous formulation

Duration of treatment / exposure:

4 hours

Observation period:

8 days

Number of animals:

1 male, 1 female

Details on study design:

TEST SITE
- Area of exposure: 2.5 cm x 2.5 cm, Upper third of the back or flank

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period with Lutrol and Lutrol/water (1:1)

SCORING SYSTEM: According to J. H. Draize: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. In Dermal Toxicity pp. 46 - 59, 1959. Austin, Tex.: Association of Food and Drug Officials of the United States, Texas, State Department of Health

READINGS: 30 - 60 minutes after removal of the test patches and 24 h, 48 h and 8 d after the beginning of application

ADDITIONAL EXAMINATIONS: After sacrifice the animals are examined by gross pathology. If there are clinical signs of necroses, these are confirmed by gross-pathological examination after incision of the skin.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable because no effects were observed

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable because no effects were observed

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable because no effects were observed

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable because no effects were observed

Irritant / corrosive response data:

No effects were observed 8 days after the treatment.

Other effects:

No other effects.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the present experiment, the test material does not give indication of an irritant property to rabbit skin.

Executive summary:

To assess the acute skin irritation or corrosion potential of the test substance in vivo a skin irritation test in Vienna White rabbits similar to OECD guideline 404 was performed. The test item was applied by topical occlusive application of 0.5 g of the 50% test substance solution to the upper third of the back or flank of each of two rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 24 and 48 hours, as well as 8 days after removal of the dressing.

The test item did not elicit any skin reactions at the application site at any of the observation times (all scores 0). No clinical signs were observed.

Under the conditions of the present experiment, the test material does not give indication of an irritant property to 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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

Draize test with an observation period of 8 days

Qualifier:

according to guideline

Guideline:

other: FED. REG. 38, NO. 187, § 1500.42, S. 27019 (27.09.1973)

Version / remarks:

equivalent to OECD test guideline 405

Principles of method if other than guideline:

Draize Test

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Gaukler, Offenbach/Main, Germany
- Mean Weight at study initiation: males: 2.90 kg; females: 2.87 kg
- Housing: one animal per cage, stainless steel with wire mesh walk floors, floor area: 40 cm x 51 cm,
- Diet (e.g. ad libitum): Ovator Solikanin 4 mm; Muskator-Werke, Duesseldorf, Germany (about 130 g per animal per day)
- Water (e.g. ad libitum): About 250 ml per animal per day: demineralized water (Monday - Friday), tap water (Saturday/Sunday)
- Acclimation period: About 8 days before the beginning of the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 26
- Humidity (%): Conventional (about 30 - 70)
- Photoperiod (hrs dark / hrs light): 12 h/12 h

Vehicle:

unchanged (no vehicle)

Controls:

other: Untreated eye

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml bulk volume (about 75 mg of the triturated test substance)/undiluted

Duration of treatment / exposure:

unspecified; the substance was not washed out

Observation period (in vivo):

1 h, 24 h, 48 h, 72 h and 8 d after application

Number of animals or in vitro replicates:

6 (3 males, 3 females)

Details on study design:

APPLICATION SITE
- Single application to the conjunctival sac of the eyelid

REMOVAL OF TEST SUBSTANCE
- Washing (if done): the substance is not washed out

SCORING SYSTEM: According to J. H. Draize: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. In Dermal Toxicity pp. 46 - 59, 1959. Austin, Tex.: Association of Food and Drug Officials of the United States, Texas, State Department of Health

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: No effects in any animal

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

other: No effects in any animal

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 8 d

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.2

Max. score:

4

Reversibility:

fully reversible within: 72 h

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

3

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 8 days

Irritation parameter:

conjunctivae score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 72h

Irritation parameter:

conjunctivae score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 72h

Irritation parameter:

conjunctivae score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 72h

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritation parameter:

chemosis score

Basis:

other: animals # 1,2,4,5,6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Other effects:

Discharge was observed in one animal at readings 24-48h (score 2) and 72h (score 1). This was reversible within 8 days.

Table 1: Irritation results

Animal	Reading	Cornea		Iritis	Conjunctiva
		Opacity	Area		Redness	Chemosis	Symptoms
1	24 h	0	0	0	0	0
2	24 h	0	0	0	0	0
3	24 h	0	0	0	2	2	RE
4	24 h	0	0	0	1	0
5	24 h	0	0	0	1	0
6	24 h	0	0	0	1	0
1	48 h	0	0	0	0	0
2	48 h	0	0	0	1	0
3	48 h	0	0	0	2	1	RE
4	48 h	0	0	0	0	0
5	48 h	0	0	0	0	0
6	48 h	0	0	0	0	0
1	72 h	0	0	0	0	0
2	72 h	0	0	0	1	0
3	72 h	0	0	0	1	0	RE
4	72 h	0	0	0	0	0
5	72 h	0	0	0	0	0
6	72 h	0	0	0	0	0
1	8 d	0	0	0	0	0
2	8 d	0	0	0	0	0
3	8 d	0	0	0	0	0	RE
4	8 d	0	0	0	0	0
5	8 d	0	0	0	0	0
6	8 d	0	0	0	0	0
Mean animal 1	24 - 72 h	0	0	0	0	0
Mean animal 2	24 - 72 h	0	0	0	0.67	0
Mean animal 3	24 - 72 h	0	0	0	1.67	1
Mean animal 4	24 - 72 h	0	0	0	0.3	0
Mean animal 5	24 - 72 h	0	0	0	0.3	0
Mean animal 6	24 - 72 h	0	0	0	0.3	0
Mean animal 1 - 6	24 - 72 h	0	0	0	0.7	0.2

RE =small retractions in the eyelids

Interpretation of results:

GHS criteria not met

Conclusions:

Considering the described ocular reactions as well as the average score for irritation, the test item is not irritating to the rabbit eye.

Executive summary:

In a primary eye irritation test the acute eye irritation or corrosion potential of the test substance was assessed by a single ocular application of 0.1 mL bulk volume (about 75 mg) of the unchanged test substance to each one eye of three Vienna White rabbits according to Fed. Reg. 38, No. 187, which is equivalent to OECD test guideline 405. The test item was not washed out. The ocular reactions were assessed approximately 1, 24, 48 and 72 hours as well as 8 days after application.

Slight or moderate conjunctival redness was observed in some animals during the course of the study, while slight to moderate conjunctival chemosis and slight to moderate discharge only were observed in one animal.

The ocular reactions were reversible in all animals within 8 days after application at the latest. The average score (24 to 72 hours) for irritation was calculated to be 0.0 for corneal opacity and for iris, 0.55 for conjunctival redness and 0.16 for chemosis. Discharge was observed in one animal at readings 24 - 48h (score 2) and 72h (score 1). This was reversible within 8 days

Considering the described ocular reactions as well as the average score for irritation, the test item is not 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

There are reliable studies available to assess the potential of the test substance for skin and eye irritation.

 

Key study: Skin irritation/corrosion

To assess the acute skin irritation or corrosion potential of the test substance in vivo a skin irritation test in Vienna White rabbits similar to OECD guideline 404 was performed (BASF SE, 1981). The test item was applied by topical occlusive application of 0.5 g of the 50% test substance solution to the upper third of the back or flank of each of two rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 24 and 48 hours, as well as 8 days after removal of the dressing.

The test item did not elicit any skin reactions at the application site at any of the observation times (all scores 0). No clinical signs were observed.

Under the conditions of the present experiment, the test material does not give indication of an irritant property to rabbit skin.

 

Key study: Eye irritation

In a primary eye irritation test the acute eye irritation or corrosion potential of the test substance was assessed by a single ocular application of 0.1 mL bulk volume (about 75 mg) of the unchanged test substance to each one eye of three Vienna White rabbits according to Fed. Reg. 38, No. 187, which is equivalent to OECD test guideline 405 (BASF SE, 1981). The test item was not washed out. The ocular reactions were assessed approximately 1, 24, 48 and 72 hours as well as 8 after application.

Slight or moderate conjunctival redness was observed in some animals during the course of the study, while slight to moderate conjunctival chemosis and slight to moderate discharge only were observed in one animal.

The ocular reactions were reversible in all animals within 8 days after application at latest. The average score (24 to 72 hours) for irritation was calculated to be 0.0 for corneal opacity and for iris, 0.55 for conjunctival redness and 0.16 for chemosis. Discharge was observed in one animal at readings 24 - 48h (score 2) and 72h (score 1). This was reversible within 8 days

Considering the described ocular reactions as well as the average score for irritation, the test item is not irritating to the rabbit eye.

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. A OECD 404 study is available for skin irritation. A study equivalent to OECD 405 is available for eye irritation. In both studies, the scores for the test item treated tissues were below the thresholds for classification as an irritant. As a result, the substance is not considered to be classified for skin or eye irritation under Regulation (EC) No. 1272/2008, as amended for the seventeenth time in Regulation (EC) No. 2021/849.