Reaction products of 7-amino-4-hydroxynaphthalene-2-sulfonic acid reacted with 2-((1,3,5-triazin-2-yl)amino)ethan-1-ol, coupled with diazotised 3-amino-4-hydroxybenzenesulfonic acid, subsequently coupled with diazotised 2-amino-5-methoxybenzenesulfonic acid, chelated with copper, sodium salt

Endpoint summary

Description of key information

Testing for primary dermal irritation or eye irritation showed that the substance is non-irritant to the skin and eye, respectively.

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:

1991-04- 30 to 1991-05-07

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:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

New Zealand White rabbit (Safepharm Definitive Protocol Number TX 3076)
Source: (David percival Ltd. Moston, Sandbach, Cheshire, U.K.)
Identification: Numbering
Husbandry: Individually housed in suspended metal cages.
-free access to food to mains drinking water and food.
3 animals (only with a healthy intact epidermis were selected for the study)
2 females, 1 male
2.25 – 2.50 kg weight
approximately. 12 – 16 weeks old
Minimum acclimatisation period of 5 days
The animal room was maintained at a temperature of 17 -21°C and relative humidity of 48-58%.
The rate of air exchange was approximately 15 changes per hour and the lighting was controlled by a time switch to give 12 hours light and 12 hours darkness.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Remarks:

On the day of the test rabbits were clipped free of fur from the dorsal/flank area suitable test site selected on the back of each rabbit.

Vehicle:

water

Remarks:

0.5 mL distilled water to form a paste

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5g of the test material, moistened with 0.5 mL of distilled water, Introduced under a 2.5 cm x 2.5 cm gauze

Duration of treatment / exposure:

4 hours

Observation period:

Approximately one hour following the removal of the patches, and 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the Draize scale J.H. (1959) Association of food and Drug Officials of the United States, Austin, Texas, 'The Appraisal of the Safety of Chemicals 1n Foods, Drugs and Cosmetics".

Reversibility of skin reactions were additionally examined on day 7

Number of animals:

3 animals (only with a healthy intact epidermis were selected for the study)
2 females, 1 male
2.25 – 2.50 kg weight
Approximately. 12 – 16 weeks old

Details on study design:

TEST SITE
0.5g of the test material, moistened with
0.5 mL of distilled water, Introduced under a 2.5 cm x 2.5 cm gauze
patch and placed in position on shorn skin. The patch was secured
in position with a strip of surgical adhesive tape {BLENDERH: approximate-
size 2.5 cm x 4.0 cm}.
To prevent the animals interfering with the
patches, the trunk of each rabbit was wrapped in an elasticated corset (TUBIGRIP)

REMOVAL OF TEST SUBSTANCE

4 hours after application the corset and patches were removed from each animal and
any residual test material removed by gentle swabbing with cotton wool soaked in distilled water followed by diethyl ether.


SCORING SYSTEM: Draize scale

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

female N0 154 (mean score 0.0)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

Male No 155 (mean score 1.0)

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

no indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

Female N0 160 (mean score 0.3)

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

animal: 1, 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Very slight erythema was seen at 2 treated skin sites at hour 24 and at one treated skin site at hour 48 and 72 of observation.
Desquamation was confined to one treated skin site at the 7 day observation.
The slight purple staining of the skin did not interfere with the evaluation of the test site.

EVALUATION OF SKIN REACTIONS

Erythema and Eschar formation                                                   Value            

No erythema 0

Very slight erythema (barely perceptible) 1

Well defined erythema 2

Moderate to severe erythema 3

Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4

 

Oedema Formation                                                                                   Value

No oedema 0

Very slight oedema (barely perceptible) 1

Slight oedema (edges of area well-defined by definite raising) 2

Moderate oedema (raised approximately 1 millimetre) 3

Severe oedema (raised more than 1 millimetre and extending

beyond the area of exposure) 4  

		Individual scores – rabbit no. and sex (bodyweight kg)
Skin reaction	Observation time	154 f (2,25)	155 m (2,5)	160 f (2,37)	Total
Erythema / Eschar Formation	1 h	0 STA	0 STA	0 STA	(0)
	24 h	0 STA	1 STA	1 STA	2
	48 h	0	1 STA	0 STA	(1)
	72 h	0	1 STA	0 STA	1
	7 d	0	0 STAD	0 STA	(0)
Oedema Formation	1 h	0	0	0	(0)
	24 h	0	0	0	0
	48 h	0	0	0	(0)
	72 h	0	0	0	0
	7 d	0	0	0	(0)
Sum of 24 – 72 h readings (S)	3
Primary Irritation index (S/6)	3/6 = 0.5
Classification	Mild Irritant ( ): total values not used for calculations of primary irritation index STA: faint purple/red-coloured staining   Individual daily and individual mean scores for dermal irritation following 4 h exposure required for EEC labelling regulations       Individual scores rabbit no. and sex (Bodyweight kg) Skin reaction Reading (hours) 154 female (2.25) 155 male (2.5) 160 (2.37) Erythema/Escher Formation 24 0 1 1 48 0 1 0 72 0 1 0 Total   0 3 1 Mean score   0.0 1.0 0.3 Odema Formation 24 0 0 0 48 0 0 0 72 0 0 0 Total   0 0 0 Mean score   0.0 0.0 0.0

 

 

Interpretation of results:

GHS criteria not met

Conclusions:

Very slight transient reddening was seen in 2 of 3 rabbits. Hence, Direct Red 83:1 is not a skin irritating substance.

Executive summary:

The aim of this study was to obtain information on the irritant effect of Direct Red 83:1 on the skin after a single dermal administration.

In a 4 hours skin irritation/corrosion study, 500 mg of Direct Red 83:1, moistened with 500 mL of distilled water, was introduced under a 2.5 cm x 2.5 cm gauze patch and placed in position with a strip of surgical adhesive tape; and was applied in the position of the shorn skin of 3 healthy young adult new Zealand white Rabbits under semi occlusive conditions.

Four hours after application the corset and patches were removed from each animal and any residual test material removed by gentle swabbing with cotton wool soaked in distilled water followed by diethyl ether.

Approximately one hour following the removal of the patches, and 24,48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the Draize scale J.H. (1959) Association of food and Drug Officials of the United States, Austin, Texas, 'The Appraisal of the Safety of Chemicals 1n Foods, Drugs and Cosmetics":

An additional observation was made on day 7 to asses reversibility of skin reactions

 

The test substance produced very slight erythema at 2 treated skin sites at hour 24 and at one treated skin site at hour 48 and 72 of observation. Desquamation was confined to one treated skin site at the 7 day observation.

No corrosive effects were noted. 

Test material was classified as non-irritant according to CLP-labelling regulations. No symbol and risk phrase are required.

Testing of Direct Red 83:1 for primary dermal irritation showed that the substance is non-irritant to the skin.

Results Synopsis

Test Substance	Test animals	ENVIRONMENTAL CONDITIONS	Control	Results	Conclusion (based on the test conditions)
Direct Red 83:1	New Zealand albino rabbit	- Temperature (°C): 17 -21 - Relative humidity: 48 -58% - Air changes (per hr): at least 15 changes - Photoperiod (hrs dark / hrs light): 12 hours daily	patch (the substance was administered over the whole surface of 2.5 x 2.5 cm cellulose patch on a piece of surgical plaster)	Faint purple/red-coloured staining was commonly noted during the study. This did not affect evaluation of skin response.   The test substance produced very slight erythema at 2 treated skin sites at hour 24 and at one treated skin site at hour 48 and 72 of observation. Desquamation was confined to one treated skin site at the 7 day observation.	Direct Red 83:1 is non-irritant to skin. No symbol and risk phrase are required.

 

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:

1991-05-19 to 1991-05-21

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

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

New Zealand White rabbit (Safepharm Definitive Protocol Number TX 3077)
Source: (David Percival Ltd. Moston, Sandbach, Cheshire, U.K.)
Identification: Numbering
Husbandry: Individually housed in suspended metal cages.
-free access to food to mains drinking water and food.
3 animals (only with a healthy intact epidermis were selected for the study)
2.96 – 3.1 kg
Approx. 12 – 16 weeks old
Minimum of acclimatisation period 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23 °C
- RELATIVE Humidity (%): 50-66%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12

IN-LIFE DATES: From: 1991-05-13 to 1991-05-21

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

Single application to non- irrigated to both eyes of the rabbits
Immediately before the start of the test
One rabbit initially tested: 0.1 mL of test material, approx. 98 mg

Duration of treatment / exposure:

Assessment of ocular damage/irritation was made approx. 1 h and 24, 48 and 72 h following the treatment.

Observation period (in vivo):

Additional observation on day 7 to assess reversibility of ocular effects.

Number of animals or in vitro replicates:

3

Details on study design:

- Time after start of exposure: 1 hour and 24, 48 and 72 hours following treatment

SCORING SYSTEM:
To classify ocular irritancy potential: modified version of system described by Kay J.H. and Calandra J.C., J. Soc. Cosmet. Chem., 1962 13 281-289
Results interpreted according to Commission Directive 83/467/EEC
TOOL USED TO ASSESS SCORE: fluorescein
Confirmation of corneal opacity was made by examination of the eye under ultra-violet illumination using sodium fluorescein B.P

Irritation parameter:

cornea opacity score

Remarks:

cornea opacity

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritation parameter:

cornea opacity score

Remarks:

cornea opacity

Basis:

animal: 2, 3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal: 1, 2

Time point:

24/48/72 h

Score:

>= 0 - <= 0.3

Max. score:

2

Reversibility:

fully reversible within: 48 h

Remarks:

at 24 h, the evaluation was impaired due to the heavy staining due to the test substance

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Conjunctival redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

>= 1.33 - <= 2

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

probability of mild irritation

Irritation parameter:

conjunctivae score

Remarks:

Conjunctival redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.6

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

Conjunctival redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Remarks:

Conjunctival chemonis

Basis:

animal: 1, 2

Time point:

24/48/72 h

Score:

1.6

Max. score:

4

Reversibility:

fully reversible within: 7 d

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Remarks:

Conjunctival chemonis

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Max. score:

3

Reversibility:

fully reversible within: 48 h

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

See results and discussions

Other effects:

None

Individual scores and individual total scores for ocular irritation

 

Rabbit No and Sex (bodyweight kg)	IPR = 3 183 male (2,96)					IPR = 0 173 Male ^+ (3,02)					IPR = 0 167 Female  ^+ (3.10)
Time after treatment	1 h	24 h	48 h	72 h	7 d	1 h	24 h	48 h	72 h	7 d	1 h	24 h	48 h	72 h	7 d
Cornea E = Degree of Opacity F = Area of Opacity	0^FS	1^FS	0	0	0	0^FS	0^FS	0	0	0	0^FS	0	0	0	0
	0	1	0	0	0	0	0	0	0	0	0	0	0	0	0
Score (ExF)x5	0	5	0	0	0	0	0	0	0	0	0	0	0	0	0
IRIS D	?s<1	?s<1	0	0	0	?s<1	?s<1	0	0	0	?s<1	0	0	0	0
Score (D x 5)	0 or 5	0 or 5	0	0	0	0 or 5	0 or 5	0	0	0	0 or 5	0	0	0	0
Conjunctiveae A = Redness B = Chemosis C = Discharge	?s	?s	2^s	2^s	0^s	?s	2^s	2^s	1^s	0^s	?s	1	0	0	0
	2	2	1	1	0	2	2	1	1	0	2	1	0	0	0
	3^Re	2^Re	1^Re	0^Re	0^Re	3^Re	1^Re	0^Re	0^Re	0^Re	3^Re	0^Re	0^Re	0^Re	0^Re
Score (A + B + C) x 2	10 - 16	8 - 14	8	6	0	10 - 16	10	6	4	0	10 – 16	4	0	0	0
Total score	10 - 32	13 - 24	8	6	0	10 - 21	10 - 15	6	4	0	10 - 21	4	0	0	0

Individual total scores and group mean scores for ocular irritation at different times of observation:

 

Rabbit Number and Sex	1 h	24 h	48 h	72 h	7 d
183 m	10 – 21	13 – 24	8	6	0
173 m	10 – 21	10 - 15	6	4	0
167 f	10 – 21	4	0	0	0
Group Total	30 – 63	27 – 43	14	10	0
Group mean score	10 – 21 1 hour	9,0 – 14.3 24 hours	4.7 48 hours	3.3 72 hours	0 7 days

 

Individual and mean scores for cornea, iris and conjunctivae required for EEC labelling Regulations

 

Rabbit Number, sex, bodyweight	Time after treatment	Corneal opacity	Iridial inflammation	Conjunctival redness	Conjuntival Chemosis
183 m (2.96)	24 hours	1	<=1	?s	2
	48 hours	0	0	2	1
	72 hours	0	0	2	1
Total		1	<=1	4-7	4
Mean		0.3	<=0.3	1.3-2.3	1.3
173 m (3.02)	24 hours	0	<1	2	2
	48 hours	0	0	2	1
	72 hours	0	0	1	1
Total		0	<1	5	4
Mean		0.0	<=0.3	1.7	1.3
167 f (3.10)	24 hours	0	0	1	1
	48 hours	0	0	0	0
	72 hours	0	0	0	0
Total		0	0	1	1
Mean		0.0	0.0	0.3	0.3

 Produced moderate conjunctival irritation. Diffuse corneal opacity was confined to one treated eye. All treated eyes appeared normal seven days after treatment.

 

 

 

 

Interpretation of results:

GHS criteria not met

Conclusions:

The test material did not produce positive criteria in any rabbit according to the CLP labeling regulation and was classified as non-irritant. No symbol and risk phrase are therefore required.

Executive summary:

In an eye irritation/corrosion study, 98 mg of Direct Red 83:1 was administered once to the non-irrigated right eye of three rabbits. In each case the untreated eyes served as a control.

The aim of this study was to assess the irritancy potential of the test substance after a single administration to the rabbit eye.

A single application of the test substance to the non-irrigated eye of three rabbits produced a transient slight to moderate conjunctival irritation. Diffuse corneal opacity was confined to one treated eye. All treated eyes appeared normal seven days after treatment.

Diffuse corneal opacity was confined to one treated eye at the 24-hour observation. No other adverse corneal effects were noted. Corneal opacity was confirmed using sodium fluorescein B.P. in all treated eyes one hour after treatment and in two treated eyes at the 24-hour observation.

Red-coloured staining precluded accurate evaluation of iridial inflammation in all treated eyes one hour after treatment and in two treated eyes at the 24-hour observation. The reaction were considered to be less than or equal to grade 1.

Red coloured standing prevented accurate evaluation of conjunctival redness in all treated eyes one hour after treatment and in one treated eye in the 24-hour observation.

Conjunctival chemosis and discharge were noted in all treated eyes one hour after treatment with slight or moderate conjunctival chemosis and discharge in one treated eye at the 24-hour observation. Minimal or moderate conjunctival reddening was noted in two treated eyes at the 24-hour observation with minimal conjuctival reddening at the 48 and 72-hour observations. Moderate conjunctival reddenind was noted in one treated eye at the 48 hour observation with minimal conjunctival reddening at the 72 -hour observation. Residual test material was noted around all treated eyes during the study.

Direct Red 83:1 did not produce positive criteria in any rabbit according to the CLP labeling regulation and was classified as non-irritant. No symbol and risk phrase are required.

 

 

 

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

In a 4 hours skin irritation/corrosion study, 500 mg test substance, moistened with 500 mL of distilled water, was introduced under a 2.5 cm x 2.5 cm gauze patch and placed in position with a strip of surgical adhesive tape; and was applied in the position of the shorn skin of 3 healthy young adult new Zealand white Rabbits under semi occlusive conditions.

Four hours after application the corset and patches were removed from each animal and any residual test material removed by gentle swabbing with cotton wool soaked in distilled water followed by diethyl ether.

Approximately one hour following the removal of the patches, and 24,48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored according to the Draize scale J.H. (1959) Association of food and Drug Officials of the United States, Austin, Texas, 'The Appraisal of the Safety of Chemicals 1n Foods, Drugs and Cosmetics":

An additional observation was made on day 7 to asses reversibility of skin reactions

The test substance produced very slight erythema at 2 treated skin sites at hour 24 and at one treated skin site at hour 48 and 72 of observation. Desquamation was confined to one treated skin site at the 7 day observation.

No corrosive effects were noted. 

Test material was classified as non-irritant according to CLP-labelling regulations. No symbol and risk phrase are required.

Testing for primary dermal irritation showed that the substance is non-irritant to the skin.

In an eye irritation/corrosion study, 98 mg of the test substance was administered once to the non-irrigated right eye of three rabbits. In each case the untreated eyes served as a control.

The aim of this study was to assess the irritancy potential of the test substance after a single administration to the rabbit eye.

A single application of the test substance to the non-irrigated eye of three rabbits produced a transient slight to moderate conjunctival irritation. Diffuse corneal opacity was confined to one treated eye. All treated eyes appeared normal seven days after treatment.

The test material did not produce positive criteria in any rabbit according to the CLP labeling regulation and was classified as non-irritant. No symbol and risk phrase are therefore required.