Amines, rosin, compds. with 9-(2-carboxyphenyl)-3,6-bis(diethylamino)xanthylium chloride and disodium hydrogen bis[4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-3-hydroxy-1-naphthalenesulfonato(3-)]chromate(3-)

Endpoint summary

Description of key information

According to results of the available studies, the test item is not classified for skin and eye irritation.

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:

1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, CH-4414 Fuellinsdorf
- Weight at study initiation: 2-3 kg
- Housing: Animals were housed individually in metal cages.
- Diet: Rabbit food, NAFAG, No. 814 Tox, NAFAG AG, Gossau, SG (Switzerland), ad libitum.
- Water (e.g. ad libitum): drinking water, ad libitum.
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +-3 °C
- Humidity: 55 +/- 15 %
- Air changes: 15 per h
- Photoperiod: 12/12 h dark/light

Type of coverage:

occlusive

Preparation of test site:

other: abraded and intact

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated skin of the same animal

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

Duration of treatment / exposure:

24 h

Observation period:

7 days

Number of animals:

6 in total. 3 males and 3 females (two treatment sites per animal, one with intact skin, one with abraded skin).

Details on study design:

TEST SITE
- Clipping of the fur: 24 hours before study start, immediatly before treatment the shaven skin as slightly abraded.
- Area of exposure: approximately 6 cm2
- Type of wrap if used: impermeable material

REMOVAL OF TEST SUBSTANCE
- Time after start of exposure: The dressings were removed after 24 h.

SCORING SYSTEM: The skin reaction was assessed upon removal and during a subsequent observation period of 7 days on the basis of the evaluation scheme as described in OECD guideline 404 (2002).

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

male (intact skin)

Time point:

other: 24, 48, 72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

male (intact skin)

Time point:

other: 24, 48, 72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

male (intact skin)

Time point:

other: 24, 48, 72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #4

Remarks:

female (intact skin)

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #5

Remarks:

female (intact skin)

Time point:

other: 24, 48, 72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

erythema score

Basis:

animal #6

Remarks:

female (intact skin)

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

male (intact skin)

Time point:

other: 24, 48, 72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

male (intact skin)

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

male (intact skin)

Time point:

other: 24, 48, 72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #4

Remarks:

female (intact skin)

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #5

Remarks:

female (intact skin)

Time point:

other: 24, 48, 72 h

Score:

0.66

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritation parameter:

edema score

Basis:

animal #6

Remarks:

female (intact skin)

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

None of the single animals exceeded an erythema or edema score of 2.

Additional observation made on abraded skin

Irritation parameter	Basis	Time point	Score	Max. score	Reversibility
erythema score	mean male (abraded skin)	24, 48, 72 h	0.33	4	reversible within: 72 h
erythema score	mean female (abraded skin)	24, 48, 72 h	0.77	4	reversible within: 7 days
edema score	mean male (abraded skin)	24, 48, 72 h	0.33	4	reversible within: 72 h
edema score	mean female (abraded skin)	24, 48, 72	0.77	4	reversible within: 7 days

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:

1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

not specified

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, CH-4414 Fuellinsdorf
- Weight at study initiation: 2-3 kg
- Housing: Animals were housed individually in metal cages.
- Diet: Rabbit food, NAFAG, No. 814 Tox, NAFAG AG, Gossau, SG (Switzerland), ad libitum.
- Water (e.g. ad libitum): drinking water, ad libitum.
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +-3 °C
- Humidity: 55 +/- 15 %
- Air changes: 15 per h
- Photoperiod: 12/12 h dark/light

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1

Duration of treatment / exposure:

Following administration, the lids were gently closed for 15 seconds.
In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of steile pysiological saline.

Observation period (in vivo):

The eyes were examined 24, 48, 72 hours and 4 and 7 days after treatment.

Number of animals or in vitro replicates:

6 in total. 3 males (unrinsed) and 3 females (rinsed).

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: For three females of the six test animals, the treated eyes were flushed with saline 30 seconds after treatment.

SCORING SYSTEM: : The eye irritation scores were evaluated according to the scoring system by OECD and Draize (1977)

TOOL USED TO ASSESS SCORE: hand-slit lamp

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Remarks on result:

other: unrinsed

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

female

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Remarks on result:

other: rinsed

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

2

Remarks on result:

other: unrinsed

Irritation parameter:

iris score

Basis:

mean

Remarks:

female

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

2

Remarks on result:

other: rinsed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Remarks:

male

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: unrinsed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Remarks:

male

Time point:

other: 24, 48, 72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: unrinsed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Remarks:

male

Time point:

other: 24, 48, 72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: unrinsed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #4

Remarks:

female

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

3

Remarks on result:

other: rinsed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #5

Remarks:

female

Time point:

other: 24, 48, 72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: rinsed

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #6

Remarks:

female

Time point:

other: 24, 48, 72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: rinsed

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 24, 48, 72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: unrinsed

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: unrinsed

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 24, 48, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 4 days

Remarks on result:

other: unrinsed

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: 24, 48, 72 h

Score:

0

Max. score:

4

Remarks on result:

other: rinsed

Interpretation of results:

GHS criteria not met

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation in vivo

Key study

The skin irritation in vivo study with New Zealand white rabbits was conducted similar to the OECD guideline 404. Three male and three female animals were treated with and 0.5 g of the test item on two different sites (on intact skin and on an abraded skin) with a treatement surface of approximately 6 cm² under an occlusive dressing. The skin reaction was assessed upon removal and during a subsequent observation period of 7 days on the basis of the evaluation scheme as described in OECD guideline 404 (2002).

The test item caused slight eythema and edema at the treatment sites which were fully reversible within 7 days. For all treated animals no scoring result for erythema and edema exceeded a value of 2.

Supporting study

The skin irritation in vivo study with New Zealand white rabbits was conducted similar to the OECD guideline 404. Three male and three female animals were treated with and 0 .5 g of the test item dissloved in polyethylene glycol (50/50 % w/v) on two different skin sites (on intact skin and on an abraded skin) with a treatement surface of approximately 2.5 x 2.5 cm under an occlusive dressing. The skin reaction was assessed upon removal and during a subsequent observation period of 7 days on the basis of the evaluation scheme as described in OECD guideline 404 (2002).

The test item did not cause any noticable eythema and edema at the treatment sites. For all animals, the individual scoring result values for erythema and edema were 0.

Eye irritation in vivo

Key study

The eye irritation in vivo study with New Zealand white rabbits was conducted similar to the OECD guideline 405. Three male and three female animals were treated with 0.1 g of the test item. 30 seconds after the application, the eyes of the female animals were rinsed with 10 mL of lukewarm saline solution, while the eyes of the male animals were left rinsed. The eyes were examined at 1, 24, 48 and 72 hours after the application with the aid of a slit lamp and effects were rated according to the draize scoring system. The test item caused slight conjunctival irritant effects which were fully reversible within 7 days. More specifically, the conjunctiva redness scores for 24 h, 48 h and 72 h observation endpoint were 1, 1.33 and1.33 for the three treated male animals, and 0, 0.33 and 0.33 for the three treated female animals, respectively.The corresponding chemosis scores were 1, 1, and 0 for the three treated male animlas and 0,0 and 0 for the three treated female animals respectively. All cornea and iris scores were 0.

Supporting study.

Similarly to the key study, an eye irritation in vivo study was conducted with English Silver strain rabbits. Again, three male and three female animals were treated with 0 .1 g of the test item. 30 seconds after the application, the eyes of the female animals were rinsed with 10 mL of lukewarm saline solution, while the eyes of the male animals were left rinsed. The eyes were examined at 1, 24, 48 and 72 hours after the application with the aid of a slit lamp and effects were rated according to the draize scoring system. The test item did not cause any elevated scores (all scores were 0).

Justification for classification or non-classification

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

The available experimental in-vivo test data are reliable and suitable for classification purposes under Regulation 1272/2008. All scores were below the thresholds identifying a substance as an irrititant. 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 sixth time in Regulation EC 944/2013.