2-[4-[[2-(cyanoimino)hexahydro-4,6-dioxo-5-pyrimidyl]azo]phenyl]-6-methylbenzothiazole-7-sulphonic acid, compound with 2,2',2''-nitrilotris[ethanol] (1:1)

Endpoint summary

Description of key information

Skin irritation:

The test item was found to be non-irritating to the rabbits' skin.

Eye irritation:

The test item was found to be non-irritating to the rabbits' eyes.

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:

1990-02-06 to 1990-02-09

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1981

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

1984

Deviations:

no

GLP compliance:

no

Specific details on test material used for the study:

- Lot/batch No.: Op. 604/89

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. K. Thomae GMBH, Chemisch-pharmazeutische Fabrik, Biberach/Riss, Germany
- Age at study initiation: 9-13 weeks
- Weight at study initiation: 2340 to 2670 g
- Housing: individually
- Diet: ad libitum standard rabbit pellet - Nafag No. 814, Gossau, Switzerland
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3 °C
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

VEHICLE
- Amount applied: Destilled water was used to moisten a gauze patch.

Duration of treatment / exposure:

4 hours

Number of animals:

3 male animals

Details on study design:

TEST SITE
- Area of exposure: 12-16 cm2

SCORING SYSTEM:
Skin reactions were evaluated according to the OECD scoring system.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24 h/ 48 h/ 72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 24 h/ 48 h/ 72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 24 h/ 48 h/ 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24 h/ 48 h/ 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 24 h/ 48 h/ 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 24 h/ 48 h/ 72 h

Score:

0

Max. score:

4

Other effects:

A substance related orange-staining was observed in all animals until 24 hours after removal of the patch.

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

Study period:

1990-02-13 to 1990-02-20

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

1981

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Version / remarks:

1984

Deviations:

no

GLP compliance:

no

Specific details on test material used for the study:

- Lot/batch No.: Op. 604/89

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Dr. K. Thomae GMBH, Chemisch-pharmazeutische Fabrik, Biberach/Riss, Germany
- Age at study initiation: 9-13 weeks
- Weight at study initiation: 2460 to 2850 g
- Housing: individually
- Diet: ad libitum standard rabbit pellet - Nafag, No. 814, Gossau, Switzerland
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20+/- 3
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 mL (43 mg)

Duration of treatment / exposure:

single application, eyelids were held together for one second to prevent loss of the article

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3

Details on study design:

SCORING SYSTEM: Ocular reactions were evaluated according to the OECD scoring system.

TOOL USED TO ASSESS SCORE: hand-slit lamp

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 24 h /48 h /72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 24 h /48 h /72 h

Score:

0

Max. score:

4

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: 24 h /48 h /72 h

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 24 h /48 h /72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 24 h /48 h /72 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: 24 h /48 h /72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24 h /48 h /72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24 h /48 h /72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: 24 h /48 h /72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24 h /48 h /72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24 h /48 h /72 h

Score:

0

Max. score:

4

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 24 h /48 h /72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 h

Interpretation of results:

not irritating

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

Key study

The substance was tested for its’ irritating property according to OECD 404 on the skin of three male rabbits. Therefore, their flanks were shaved 24 hours before treatment. A gauze patch moistened with distilled water and bearing 0.5 g of the test article was applied to one flank, as control a gauze patch moistened with water was applied to the other flank. The patches were covered with aluminum foil. After an exposure time of 4 hours, the patches were removed and skin reactions were evaluated after 1, 24, 48 and 72 hours. As a result, the mean erythema scores over the time points 24 h/ 48 h/ 72 h were found to be 0 for all animals as well as the edema mean scores. As no skin reactions were found, the test was terminated after 72 hours.

Supporting study no. 1:

 In a second skin irritation study, the test compound was tested on three rabbits. 0.5 mL was placed on a gauze patch (20 cm^2) which was applied to the shaved flank and loosely covered with aluminum foil. A control patch was applied to the other flank. The exposure duration was 4 hours. The animals were checked daily for systemic symptoms and mortality. The skin reactions were evaluated 1, 24, 48, and 72 hours after removing the gauze patches. Because intensive staining was observed, the observation period was extended to 7 days. An exact erythema evaluation was not possible in two of the three animals due to the staining. The third animal did not exhibit erythema at any time. The mean edema scores for animals 1, 2 and three were 0, 0, and 0 respectively. The test material was therefore concluded to be not irritating to skin. This result can be regarded as supporting information, as the test substance composition included only 10 % of the active compound.

Supporting study no. 2:

A further skin irritation study was conducted on six rabbits testing on both intact and abraded skin sites. 0.5 mL of the test compound were applied to each test site on a 2.5 cm square gauze pad covered with aluminum foil and held in place by a self-adhesive bandage. After 24 hours the dressing was removed and the intact and abraded application sites were assessed for edema and erythema. No erythema formation was observed. Edema formation was found after 24 hours on the intact skin sites of two animals and on the abraded skin sites of three animals. These findings were reversible within 72 hours. Therefore, the study was terminated after 72 hours. The mean score over the times 24 h and 72 h was 0 for erythema, including both the results found on intact and abraded skin. The mean edema score on intact skin was 0.17 and the mean edema score on abraded skin was 0.25. But, as the composition of the test material is unknown, these results were not used for assessment.

Eye irritation

Key study

The test material was tested for eye irritating properties according to OECD 405 using three female rabbits. 0.1 mL (43 mg) of the test substance was placed into the conjunctival sac of one eye of each animal, the other eye served as control. Ocular reactions were evaluated and recorded after 1, 24, 48 and 72 hours after application, using the OECD scoring system. The following mean values (scores after 24, 48 and 72 hours) were recorded: cornea scores: 0 for all animals, iris: 0 for all animals, conjunctiva redness: 1.33, 0.33 and 1 for animals 1, 2 and 3, respectively, chemosis: 0.33, 0, and 0.33 for animals 1, 2 and 3. Based on these results, the test material was determined to be non-irritating to the rabbits’ eyes.

Supporting study no.1

In a second eye irritation study, the test compound was tested on three rabbits. 0.1 mL was placed into the conjunctival sac of the right eye of each animal, after gently pulling away the lower lid from the eyeball. The lids were then held together for about one second in order to prevent loss of the test material. The left eye remained untreated and served as a control. The animals were checked daily for systemic symptoms and mortality. As a result, the mean scores for all animals over the time points 24 h, 48 h, 72 h were 0 for cornea, 0 for iris, 0 for conjunctivae and 0 for chemosis. The test material was therefore concluded to be not irritating to eyes. This result can be regarded as supporting information, as the test substance composition included only 10 % of the active compound.

Supporting study no. 2

Another eye irritation study was conducted on six rabbits. 0.1 mL of the test compound was instilled into the conjunctival sac of the left eye. The eyelids were then held closed for 1 second. The right eye served as a control. After 30 seconds the compound was, as far as possible, flushed out of the eyes of three of the rabbits with approximately 200 mL of warm water. The rabbits were examined 1, 24, 48 and 72 hours after application of the test compound. A further period was not considered necessary. Very slight reactions were seen in 3/3 unwashed eyes and 1/3 washed eyes one hour after compound application. 2/3 unwashed eyes and 1/3 washed eyes were normal at 24 hours and the remaining unwashed eye by 48 hours. Slight cornea opacity with damage to the surface epithelium was seen in 2/3 unwashed eyes at one hour only. The reactions in unwashed eyes were slightly more severe than those in washed eyes. The mean scores over the times 24 h, 48 h and 72 h were 0 for cornea, 0 for iris, 0.06 for conjunctivae redness and 0 for chemosis. But, as the composition of the test material is unknown, these resultswere not used for assessment.

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. The substance does not show a skin irritation potential in the in vivo skin irritation and corrosion test and does not show an eye irritation potential in the in vivo eye irritation test. As a result the substance is not considered to be classified for skin irritation/corrosion or eye irritation/ serious eye damage under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.