Sodium bis[2-chloro-4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-5-hydroxybenzenesulphonamidato(2-)]chromate(1-)

Endpoint summary

Description of key information

Acid Red 211 is considered to be non-irritant to the rabbit skin and as an irritant to the eyes of rabbits.

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:

October 1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: "Hazardous Substances Regulations" under the U.S. Federal Hazardous Substances Labelling Act Sect. 191.11 (February 1965).

Deviations:

not specified

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Inhouse
- Age at study initiation: 12 - 16 weeks
- Weight at study initiation: Male 2.93 kg and Female 2.78 kg
- Housing: caged singly
- Diet: A commercial irradiated diet (Styles-Oxoid) was fed ad lib ad libitum
- Water: Sterile filtered water ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 1
- Humidity (%): 50-70
- Photoperiod (hrs dark / hrs light): 10 h light

Type of coverage:

occlusive

Preparation of test site:

shaved

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

FAT 20028/A was applied to each side in quantities of 0.5 g.

Duration of treatment / exposure:

The gauze patches were removed 24 h after the application.

Observation period:

24 -72 h

Number of animals:

6 rabbits (3 males and 3 females)

Details on study design:

Twenty-four hours prior to the dermal application, the backs of the rabbits were shaved over an area consisting of at least 10 % of the total body surface. Two test sites lateral to the mid line of the back were used on each rabbit. Immediately before the application of the test compound, the right hand site was abraded with the point of a sterile hypodermic needle. The abrasions were sufficiently deep to penetrate the stratum corneum but not to damage the dermis. The left hand site remained intact. 10g of the test compound mixed with 10 mls of water to make a solution of 15 ml 0.75 ml of which was applied to each test site on a 2.5cm square gauze pad. These were covered with aluminium foil secured with "Sleek"* adhesive tape. The test sites were then covered by a 6" wide "Coban" self adhesive bandage in order to retain the test substance in close contact with the skin. After 24 h the dressing was removed and' the intact and abraded application sites were assessed for oedema and erythema. A further assessment was made at 72 h and for any further period considered necessary up to 14 days.

SCORING SYSTEM:
Evaluation of Skin Reactions

Erythema and Eschar Formation
-No erythema: 0
-Slight erythema (barely perceptible): 1
-Well defined erythema: 2
-Moderate to severe erythema: 3
-Severe erythema (beet red) to slight eschar formation: 4

Edema Formation
-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 mm.): 3
-Severe-oedema (raised more than 1 mm. and extending beyond the area of exposure): 4

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 males and 3 females

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: Intact skin

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 males and 3 females

Time point:

24 h

Score:

1.17

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: Abraded skin

Irritation parameter:

erythema score

Basis:

mean

Remarks:

6 rabbits

Time point:

48 h

Remarks on result:

not measured/tested

Remarks:

intact skin and abraded skin

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 males and 3 females

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

other: intact skin and abraded skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 males and 3 females

Time point:

24 h

Score:

2.17

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

other: Intact skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 males and 3 females

Time point:

72 h

Score:

0

Max. score:

4

Remarks on result:

other: Intact skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 males and 3 females

Time point:

24 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 6 days

Remarks on result:

other: Abraded skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 males and 3 females

Time point:

72 h

Score:

0.17

Max. score:

4

Reversibility:

fully reversible within: 6 days

Remarks on result:

other: Abraded skin

Irritation parameter:

edema score

Basis:

mean

Remarks:

6 rabbit

Time point:

48 h

Remarks on result:

not measured/tested

Remarks:

intact skin and abraded skin

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Remarks:

3 males and 3 females

Time point:

24/48/72 h

Score:

1.63

Max. score:

4

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

The primary irritation index as the measure of the acute irritation to the skin of rabbits was found to be 1.6.

Erythema Score

Animal Number	Intact skin		Abraded skin
	24 h	72 h	24 h	72 h
11	1	0	1	0
13	1	0	1	0
15	1	0	1	0
12	1	0	1	0
14	1	0	2	0
16	1	0	1	0
Mean	1	0	1.17	0

 

Edema Score

Animal Number	Intact skin		Abraded skin
	24 h	72 h	24 h	72 h
11	2	0	1	0
13	3	0	3	1
15	2	0	2	0
12	2	0	2	0
14	2	0	2	0
16	2	0	2	0
Mean	2.17	0	2	0.17

 

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 20028/B cosidered as to be non-irritating to rabbit skin.

Executive summary:

The purpose of this test was to determine the primary-irritation index, which serves as a measure of the acute irritation, provoked by the compound on the skin of the rabbits. The procedure adopted was the patch-test technique described in the " "Hazardous Substances Regulations" under the U.S. Federal Hazardous Substances Labelling Act Sect. 191.11 (February 1955). Six rabbits (3 males/3 females), New Zealand White (Porton strain), weighing 2.78 to 2.93 kg and aged 12 -16 weeks. Twenty-four hours prior to the dermal application, the backs of the rabbits were shaved over an area consisting of at least 10 % of the total body surface. Two test sites lateral to the mid line of the back were used on each rabbit. Immediately before the application of the test compound, the right hand site was abraded with the point of a sterile hypodermic needle. The abrasions were sufficiently deep to penetrate the stratum corneum but not to damage the dermis. The left hand site remained intact. 10 g of the test compound mixed with 10mls of water to make a solution of 15 ml 0.75 ml of which was applied to each test site on a 2.5 cm square gauze pad. These were covered with aluminium foil secured with "Sleek" adhesive tape. The test sites were then covered by a 6" wide "Coban" self-adhesive bandage in order to retain the test substance in close contact with the skin. After 24 h the dressing was removed and' the intact and abraded application sites were assessed for oedema and erythema. Slight to well defined erythema and very slight to moderate oedema were seen on all sites after 24 h while very slight oedema was seen on 1/6 abraded sites after 72 h. All other sites were normal. The remaining site was become normal 6 days. In conclusion, the primary irritation to the skin of rabbits was found to be 1.6 and FAT 20028/B cosidered as to be non-irritating 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

Study period:

October 1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965).

Deviations:

not specified

GLP compliance:

no

Species:

rabbit

Strain:

New Zealand White

Remarks:

Porton strain

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Inhouse breeding
- Age at study initiation: 12 - 16 weeks
- Weight at study initiation: Males 2-93 kg and Female 2.78 kg
- Housing: caged singly
- Diet: A commercial irradiated diet (Styles-Oxoid) was fed ad libitum
- Water: Sterile filtered water ad libitum
- Acclimation period: One week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 1
- Humidity (%): 50 - 70
- Photoperiod (hrs dark / hrs light): artificial light for 10 h

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g of the test substance.

Duration of treatment / exposure:

single application

Observation period (in vivo):

1 and 24 h and after 2, 3, 6, 10 and 14 days.

Number of animals or in vitro replicates:

6 rabbits (3 males and 3 females)

Details on study design:

The eyes of the experimental animals were examined and found normal prior to the test. 100 mg of the test compound were 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.

SCORING SYSTEM: Evaluation of the Eye Reactions.
CORNEA:
A. Opacity-degree of density (area most dense taken for reading)
-No opacity: 0
-Scattered or diffuse area, details of iris clearly visible: 1
-Easily discernible translucent areas, details of iris slightly obscured: 2
-Opalescent areas, no details of iris visible, size of pupil barely discernible: 3
-Opaque, iris invisible: 4
B. Area of cornea involved
-One quarter (or less) but not zero: 1
-Greater than one quarter, but less than half: 2
-Greater than half, but less than three quarters: 3
-Greater than three quarters, up to whole area: 4
A x B x 5 Total maximum = 80

IRIS
A. Values
-Normal: 0
-Folds above normal, congestion, swelling, circum-corneal injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive): 1
-No reaction to light, hemorrhage, gross destruction (any or all of these): 2
A x 5

CONJUNCTIVAE
A. Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
-Vessels normal: 0
-Vessels definitely injected above normal: 1
-More diffuse, deeper crimson red, individual vessels not easily discernible: 2
-Diffuse beefy red: 3

B. CHEMOSIS
-No swelling: 0
-Any swelling above normal (includes nictitating membrane): 1
-Obvious swelling with partial eversion of lids: 2
-Swelling with lids about half closed: 3
-Swelling with lids about half closed to completely closed: 4

C. DISCHARGE
-No discharge: 0
-Any amount different from normal (does not include small amounts observed in inner canthus of normal animals): 1
-Discharge with moistening of the lids and hairs just adjacent to lids: 2
-Discharge with moistening of the lids and hairs, and considerable area around the eye: 3
(A + B + C) x 2 Total maximum =20

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks on result:

other: Eyes rinsed and eyes not rinsed

Irritation parameter:

cornea opacity score

Remarks:

Area involved

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

3.55

Max. score:

4

Reversibility:

not fully reversible within: 14 d

Remarks on result:

other:

Remarks:

Eyes rinsed and eyes not rinsed

Irritation parameter:

iris score

Basis:

mean

Remarks:

Animal no 11, 12, 13, 15 and 16

Time point:

24 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 14 d

Irritation parameter:

iris score

Remarks:

Eyes not rinsed

Basis:

animal #4

Remarks:

Rabbit 14

Time point:

24 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Remarks:

Eyes rinsed

Basis:

mean

Remarks:

Rabbit 11,12,13

Time point:

48 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 14 d

Irritation parameter:

iris score

Remarks:

Eyes not rinsed

Basis:

mean

Remarks:

Rabbit 14,15,16

Time point:

48 h

Score:

0

Max. score:

2

Irritation parameter:

iris score

Remarks:

Eyes rinsed

Basis:

mean

Remarks:

Rabbit 11,12,13

Time point:

72 h

Score:

1

Max. score:

2

Reversibility:

not fully reversible within: 14 d

Irritation parameter:

iris score

Remarks:

Eyes not rinsed

Basis:

mean

Remarks:

Rabbit 14,15,16

Time point:

72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

2.11

Max. score:

3

Reversibility:

not fully reversible within: 14 d

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

1.83

Max. score:

4

Reversibility:

not fully reversible within: 14 d

Irritation parameter:

overall irritation score

Basis:

mean

Remarks:

6 rabbits

Time point:

24/48/72 h

Score:

4.44

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

The primary eye irritation score is 4.44.

Conjunctiva:

On initial application of the compound there appeared to be a moderately painful reaction. A mild to moderate conjunctival reaction was seen in all treated eyes one hour after the application of the compound. At 24 h there was very little change from the initial reaction but thereafter these started to improve until at termination of study on day 14 one washed eye was normal, 5/5 unwashed and 2/5 washed eyes were showing a very slight to slight reaction and the remaining unwashed eye a moderate reaction.

Cornea:

Slight opacity with damage to the surface epithelium was seen in all treated eyes at one hour. This was still present at 24 hours and did not vary much for up to 72 h. At 6 days the 3 washed eyes were normal but the unwashed eyes were all showing slight pannus formation. In 2 of these the pannus increased in severity slightly over the remainder of the test period and was still present at termination.

Iris:

Increased folding was seen in 3/5 unwashed and 2/5 washed eyes at 24 h. The washed eyes were normal by 48 h and one unwashed eye at 14 days. The other 2 unwashed eyes were still showing increased folding at termination. All eyes would have eventually returned to normal if sufficient time had been allowed to elapse. However, 2 out of the 3 unwashed eyes might have had some very small residual scarring of the cornea. Generally, the reactions seen in washed eyes, particularly after the initial reaction, were less severe and recovery was faster than those in unwashed eyes.

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

FAT 20028/B is considered as an irritant to the rabbit eye with the primary eye irritation index is 4.44.

Executive summary:

The study was carried out with FAT 20028/B in NZW rabbit according to the procedure adopted was the technique described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965). The substance was tested on 6 rabbits (3 males/3 females), New Zealand White rabbit (Porton strain). Only rabbits showing normal ophthalmic findings were included in the test. 100 mg of the test compound were 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 and for any further period that was considered necessary.

 

RESULTS:

Conjunctiva:

On initial application of the compound there appeared to be a moderately painful reaction. A mild to moderate conjunctival reaction was seen in all treated eyes one hour after the application of the compound. At 24 h there was very little change from the initial reaction but thereafter these started to improve until at termination of study on day 14 one washed eye was normal, 5/5 unwashed and 2/5 washed eyes were showing a very slight to slight reaction and the remaining unwashed eye a moderate reaction.

 

Cornea:

Slight opacity with damage to the surface epithelium was seen in all treated eyes at one hour. This was still present at 24 hours and did not vary much for up to 72 h. At 6 days the 3 washed eyes were normal but the unwashed eyes were all showing slight pannus formation. In 2 of these the pannus increased in severity slightly over the remainder of the test period and was still present at termination.

 

Iris:

Increased folding was seen in 3/5 unwashed and 2/5 washed eyes at 24 h. The washed eyes were normal by 48 h and one unwashed eye at 14 days. The other 2 unwashed eyes were still showing increased folding at termination. All eyes would have eventually returned to normal if sufficient time had been allowed to elapse. However, 2 out of the 3 unwashed eyes might have had some very small residual scarring of the cornea. Generally, the reactions seen in washed eyes, particularly after the initial reaction, were less severe and recovery was faster than those in unwashed eyes. Therefore, FAT 20028/B is to be considered as a minimal irritant to the eye of rabbits and the primary eye irritation index is 4.44.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation:

The purpose of this test was to determine the primary-irritation index, which serves as a measure of the acute irritation, provoked by the compound on the skin of the rabbits. The procedure adopted was the patch-test technique described in the " "Hazardous Substances Regulations" under the U.S. Federal Hazardous Substances Labelling Act Sect. 191.11 (February 1955). Six rabbits (3 males/3 females), New Zealand White (Porton strain), weighing 2.78 to 2.93 kg and aged 12 -16 weeks. Twenty-four hours prior to the dermal application, the backs of the rabbits were shaved over an area consisting of at least 10 % of the total body surface. Two test sites lateral to the mid line of the back were used on each rabbit. Immediately before the application of the test compound, the right hand site was abraded with the point of a sterile hypodermic needle. The abrasions were sufficiently deep to penetrate the stratum corneum but not to damage the dermis. The left hand site remained intact. 10 g of the test compound mixed with 10mls of water to make a solution of 15 ml 0.75 ml of which was applied to each test site on a 2.5 cm square gauze pad. These were covered with aluminium foil secured with "Sleek" adhesive tape. The test sites were then covered by a 6" wide "Coban" self-adhesive bandage in order to retain the test substance in close contact with the skin. After 24 h the dressing was removed and the intact and abraded application sites were assessed for oedema and erythema. Slight to well defined erythema and very slight to moderate oedema were seen on all sites after 24 h while very slight oedema was seen on 1/6 abraded sites after 72 h. All other sites were normal. The remaining site was become normal 6 days. In conclusion, the primary irritation to the skin of rabbits was found to be 1.6 and FAT 20028/B cosidered as to be non irritating to rabbit skin. In another two supporting studies (1974 and 1980), FAT 28200 also found non-irritating to rabbit skin.

Eye Irritation:

The study was carried out with FAT 20028/B in NZW rabbit according to the procedure adopted was the technique described in the "Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics" (1959) of the US "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965). The substance was tested on 6 rabbits (3 males/3 females), New Zealand White rabbit (Porton strain). Only rabbits showing normal ophthalmic findings were included in the test. 100 mg of the test compound were 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 and for any further period that was considered necessary.

 

RESULTS:

Conjunctiva:

On initial application of the compound there appeared to be a moderately painful reaction. A mild to moderate conjunctival reaction was seen in all treated eyes one hour after the application of the compound. At 24 h there was very little change from the initial reaction but thereafter these started to improve until at termination of study on day 14 one washed eye was normal, 5/5 unwashed and 2/5 washed eyes were showing a very slight to slight reaction and the remaining unwashed eye a moderate reaction.

 

Cornea:

Slight opacity with damage to the surface epithelium was seen in all treated eyes at one hour. This was still present at 24 hours and did not vary much for up to 72 h. At 6 days the 3 washed eyes were normal but the unwashed eyes were all showing slight pannus formation. In 2 of these the pannus increased in severity slightly over the remainder of the test period and was still present at termination.

 

Iris:

Increased folding was seen in 3/5 unwashed and 2/5 washed eyes at 24 h. The washed eyes were normal by 48 h and one unwashed eye at 14 days. The other 2 unwashed eyes were still showing increased folding at termination. All eyes would have eventually returned to normal if sufficient time had been allowed to elapse. However, 2 out of the 3 unwashed eyes might have had some very small residual scarring of the cornea. Generally, the reactions seen in washed eyes, particularly after the initial reaction, were less severe and recovery was faster than those in unwashed eyes. Therefore, FAT 20028/B is to be considered as a minimal irritant to the eye of rabbits and the primary eye irritation index is 4.44. However, in another two supporting studies FAT 28200 was not irritating to rabiit eyes.

Justification for classification or non-classification

Based on the results observed in one test on eye irritation, the substance is classified as irritating to the eyes.