Tetrasodium 7-[[4-[[4,6-bis[(3-sulfonatopropyl)thio]-1,3,5-triazin-2-yl]amino]-3-methoxyphenyl]azo]naphthalene-1,3-disulfonate

Endpoint summary

Description of key information

Skin irritation = Not irritating, rabbit, OECD 404, EU method B.4, Manciaux 1998a
Eye irritation = Not irritating, rabbit, OECD 405, EU method B.5, Manciaux 1998b

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:

16 June 1998 to 19 June 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, performed in accordance with valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.3 ± 0.3 kg (mean)
- Housing: individually in polystyrene cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): approximately 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light

IN-LIFE DATES: From 16 June 1998 to 19 June 1998

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

500 mg

Duration of treatment / exposure:

4 hours

Observation period:

3 days post administration

Number of animals:

3 males

Details on study design:

TEST SITE
- Area of exposure: On the day prior to application, the flanks of each animal were clipped using electric clippers. A dose of 500 mg of test material was applied to a 6 cm² hydrophilic gauze pad moistened with water and this was then applied to the right flank of the animals for 4 hours. The left flank did not receive any test material and served as an untreated control.
- Type of wrap if used: The test material and gauze pad were held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing and a restraining bandage.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, the dressing was removed and any residual test material was wiped off with a gauze pad moistened with water.
- Time after start of exposure: approximately 4 hours

SCORING SYSTEM:
The Draize scale, presented as table 1 in the field "Any other information on material and methods incl. tables" was used to assess the degree of erythema and oedema at the application sites approximately 1 hour and 1, 2 and 3 days after the removal of the dressings.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: overall at 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: overall at 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: overall at 24, 48 and 72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Reversibility:

other: no adverse effects observed

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Reversibility:

other: no adverse effects observed

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Reversibility:

other: no adverse effects observed

Irritant / corrosive response data:

A well-defined erythema (grade 2) was observed in two animals on days 1 and 2, then a very slight erythema (grade 1) was noted in these animals on day 3. In the third animal, only a very slight erythema (grade 1) was observed on days 1 and 2.
A slight orange colouration of the skin was noted in all animals from day 1 up to the end of the observation period (day 4).

Table 2: Individual cutaneous examination and mean values of the scores recorded at each time point for each animal

Animal no.	Dermal irritation	Scores				Mean irritation score*
		1 h	24 h	48 h	72 h
1	Erythema	2	2	1	0	1.0
	Oedema	0	0	0	0	0.0
	Other	C	C	C	C
2	Erythema	2	2	1	0	1.0
	Oedema	0	0	0	0	0.0
	Other	C	C	C	C
3	Erythema	1	1	0	0	0.3
	Oedema	0	0	0	0	0.0
	Other	C	C	C	C

* of the 24, 48 amd 72 hour scores

C = orange colouration of the skin

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the study, the test material did not elicit any reaction in any of the animals during the course of the study that meant the test material required classification as a skin irritant. Only very slight erythema was seen and this was fully reversible with 72 hours. The study is considered to be reliable, relevant and adequate for risk assessment and classification and labelling purposes.

Executive summary:

The skin irritation potential of the test material was determined in accordance with standardised guidelines OECD 404 and EU Method B.4. Three rabbits received a single four hour application of 500 mg of test material and were assessed for the following 3 days for any signs of skin irritation. The mean score over 24, 48 and 72 hours for each individual animal was 0.0 for oedema. The mean score over 24, 48 and 72 hours for each individual animal was 1.0, 1.0 and 0.3 for erythema. All skin reactions were fully reversible within 72 hours. Therefore, under the conditions of the study, the test material did not elicit any reaction in any of the animals during the course of the study that meant the test material required classification as a skin irritant.

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:

24 June 1998 to 27 June 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, perfomed in accordance with valid guidelines and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France
- Weight at study initiation: 2.4 ± 0.3 kg (mean)
- Housing: individually in polystyrene cages
- Diet: 112 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France) ad libitum
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): approximately 12 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light

IN-LIFE DATES: From 24 June 1998 to 27 June 1998

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg

Duration of treatment / exposure:

- Dose administration: Approximately 100 mg of the test material was applied into the conjunctival sac of the left eye of each rabbit by gently pulling the lower lid away from the eyeball. The lower and upper eyelids were then gently held together for about 1 second to avoid any loss of test material. The right eye remained untreated and served as a control.

Observation period (in vivo):

72 hours following dose administration

Number of animals or in vitro replicates:

3 males

Details on study design:

REMOVAL OF TEST SUBSTANCE
- The eyes were not rinsed after administration of the test material

SCORING SYSTEM
- The reactions observed were scored in accordance with the criteria of Draize (see field "Any other information on materials and methods incl. tables" for further information)

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

3

Reversibility:

other: no adverse effects observed

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

3

Reversibility:

other: no adverse effects observed

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Time point:

other: overall at 24, 48 and 72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: overall at 24, 48 and 72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: overall at 24, 48 and 72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: overall at 24, 48 and 72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Reversibility:

other: no adverse effects observed

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Reversibility:

other: no adverse effects observed

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

4

Reversibility:

other: no adverse effects observed

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

2

Reversibility:

other: no adverse effects observed

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

2

Reversibility:

other: no adverse effects observed

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: overall at 24, 48 and 72 h

Score:

0

Max. score:

2

Reversibility:

other: no adverse effects observed

Irritant / corrosive response data:

Very slight or slight conjunctival reactions were observed in all animals from day 1: very slight or slight chemosis (grade 1 or 2) and very slight redness of the conjunctiva (grade 1) were observed. These reactions persisted up to day 3 at the latest. No other ocular reactions were noted during the study.

Test animals

- Source: Elevage Cunicole de Val de Selle, 80160 Prouzel, France

- Diet: 112 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France)

Table 2: Individual ocular examination and mean values of the scores recroded at each time point for each animal

Animal no.	Region of eye	Description of ocular reaction	Scores				Mean irritation score†
			1 h	24 h	48 h	72 h
1	Conjunctivae	Chemosis	1	1	0	0	0.3
		Redness	LC	0	0	0	0.0
		Discharge	0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Corneal opacity	Intensity	0	0	0	0	0.0
		Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		N	Y	N	N
2	Conjunctivae	Chemosis	2	1	0	0	0.3
		Redness	LC	0	0	0	0.0
		Discharge	0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Corneal opacity	Intensity	0	0	0	0	0.0
		Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		N	Y	N	N
3	Conjunctivae	Chemosis	2	1	1	0	0.7
		Redness	LC	1	0	0	0.3
		Discharge	0	0	0	0	0.0
	Iris		0	0	0	0	0.0
	Corneal opacity	Intensity	0	0	0	0	0.0
		Area	0	0	0	0	0.0
	Other		*	*	*	*
	Fluorescein		N	Y	N	N

† mean of 24, 48 and 72 hour scores

* = none

N = no

Y = yes

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the study, the test material only elicited slight reactions in any of the animals during the course of the study that meant the test material does not require classification as an eye irritant. The study is considered to be reliable, relevant and adequate for risk assessment and classification and labelling purposes.

Executive summary:

The eye irritation potential of the test material was determined in accordance with standardised guidelines OECD 405 and EU Method B.5. Approximately 100 mg of test material was applied into one eye of each of three rabbits; the animals were assessed for up to 72 hours to determine the grade of ocular reaction. Under the conditions of the study, very slight or slight conjunctival reactions were observed in all animals from day 1: very slight or slight chemosis (grade 1 or 2) and very slight redness of the conjunctiva (grade 1) were observed. These reactions persisted up to day 3 at the latest. No other ocular reactions were noted during the study. The test material was therefore determined to be not-irritating to the eye.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin

The skin irritation potential of the test material was determined in accordance with standardised guidelines OECD 404 and EU Method B.4. Three rabbits received a single four hour application of 500 mg of test material and were assessed for the following 3 days for any signs of skin irritation. The mean score over 24, 48 and 72 hours for each individual animal was 0.0 for oedema. The mean score over 24, 48 and 72 hours for each individual animal was 1.0, 1.0 and 0.3 for erythema. All skin reactions were fully reversible within 72 hours. Therefore, under the conditions of the study, the test material did not elicit any reaction in any of the animals during the course of the study that meant the test material required classification as a skin irritant.

Eye

The eye irritation potential of the test material was determined in accordance with standardised guidelines OECD 405 and EU Method B.5. Approximately 100 mg of test material was applied into one eye of each of three rabbits; the animals were assessed for up to 72 hours to determine the grade of ocular reaction. Under the conditions of the study, very slight or slight conjunctival reactions were observed in all animals from day 1: very slight or slight chemosis (grade 1 or 2) and very slight redness of the conjunctiva (grade 1) were observed. These reactions persisted up to day 3 at the latest. No other ocular reactions were noted during the study. The test material was therefore determined to be not-irritating to the eye.

Both studies presented to assess the irritation and corrosion potential of the test material were performed in line with GLP and accepted standardised guidelines with a high standard of reporting. Both studies were assigned a reliability score of 1 in accordance with the criteria for assessing data quality as outlined in Klimisch (1997) and considered suitable for assessment as an accurate reflection of the test material.

The available data are considered to be complete and the conclusion for both skin irritation and corrosion and eye irritation, not irritating, was taken forward for risk assessment.

Justification for selection of skin irritation / corrosion endpoint:
Only one study is available.

Justification for selection of eye irritation endpoint:
Only one study is available.

Justification for classification or non-classification

Skin

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the test material does not require classification for skin irritation.

Eye

In accordance with the criteria for classification as defined in Annex I, Regulation (EC) No 1272/2008, the test material does not require classification for eye irritation.