Pentasodium 4-amino-6-[[5-[(4-amino-6-chloro-1,3,5-triazin-2-yl)amino]-2-sulphonatophenyl]azo]-3-[(2,5-disulphonatophenyl)azo]-5-hydroxynaphthalene-2,7-disulphonate

Endpoint summary

Description of key information

Studies carried out to evaluate skin irritation/corrosion and eye irritation potency of the test substance following OECD guidelines and GLP has demonstrated that Reactive Black 039 is non-irritant and non-corrosive to skin and irritating to eye. 

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:

From 13 October, 1992 to 16 October, 1992

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

Principles of method if other than guideline:

Guidelines followed

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: Animal #1: Approximately 13 weeks; Animal #2 and #3: Approximately 12 weeks
- Weight at study initiation: 2419 to 2664 g
- Housing: Individually in labelled cages with perforated floors and equipped with an automatic drinking system (ITl, Bergen, The Netherlands)
- Diet: Standard laboratory rabbit diet (LKK-20, pellet diameter 4 mm, Hope Farms, Woerden, The Netherlands) approximately 100 g/d, ad libitum
- Water: Tap-water diluted with decalcified water, ad libitum
- Acclimation period: At least five days

ENVIRONMENTAL CONDITIONS
- Temperature: 21 °C
- Humidity: 55 %
- Air changes: Air-conditioned with 15 air changes/h
- Photoperiod: 12 h artificial fluorescent light/12 h dark

IN-LIFE DATES: From October 13, 1992 to October 16, 1992

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Controls:

other: the untreated flank of the same animal

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 500 mg

VEHICLE
- The test substance was moistened with distilled water, immediately before application, to ensure close contact to the animal's skin.

Duration of treatment / exposure:

4 h

Observation period:

Up to 72 h (The skin reactions were assessed at approximately 45 min, 24, 48 and 72 h after the removal of the dressings and test substance)

Number of animals:

Three male animals.

Details on study design:

TEST SITE
- Area of exposure: Approx. 10x15 cm²
- Type of wrap if used: The test substance was applied using a moistened Scotchpak-non-woven patch mounted on Micropore tape (3M, St. Paul, U.S.A.). The contralateral flank was similarly prepared (but without test substance and vehicle) to act as a procedural control. The dressing was wrapped around the abdomen and secured with an elastic bandage (Coban, 3M, St. Paul, U.S.A.).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Removed at the end of the exposure period using a tissue moistened with tap water and subsequently a dry tissue.
- Time after start of exposure: 4 h

SCORING SYSTEM: Draize scoring system

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Scoring of erythema was difficult due to blue staining of the treated skin at all observations (i.e., 45 minutes, 24, 48 and 72 h)

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Scoring of erythema was difficult at 24 h due to blue staining of the treated skin by the test substance.

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Scoring of erythema was difficult at 24 h due to blue staining of the treated skin by the test substance.

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Irritant / corrosive response data:

The observed skin irritation consisted of very slight erythema in animal #2 1 h after exposure, which had resolved within 24 h. No skin irritation was observed in animal #1 and #3 during the study.
There was no evidence of a corrosive effect on the skin.

Other effects:

Colouration: Blue staining of the treated skin by the test substance was observed, which made scoring of erythema difficult at the first two observations.
Body weights: The body weights and body weight gain in the animals were normal. Variation in body weight gain may be considered the result of treatment procedures.
Toxic symptoms/mortality: No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance was considered to be a non-irritant to skin.

Executive summary:

A study was performed to assess the skin irritation potential of the test substance (of ca. 100 % purity) in New Zealand White rabbits according to OECD Guideline 404 and EU Method B.4 in compliance with GLP.

500 mg of test substance were applied to a clipped skin area of 10 x 15 cm² on one flank of each animal under a 2 x 3 cm² gauze pad. The pad was allowed to remain in contact with the skin for 4 h. Animals were scored for irritation reactions after 45 minutes, and 24, 48 and 72 h. They were monitored daily for viability, mortality and clinical signs. Body weights were recorded pre-test and at study termination.

Blue staining of the treated skin by the test substance was observed, which made scoring of erythema difficult at 45 minutes and 24 h in 2/3 animals and in 1/3 animal at all observations. The observed skin irritation consisted of very slight erythema in 1/3 animal 45 minutes after exposure, which had resolved within 24 h. No skin irritation was observed in remaining animals. No signs of body weight variability, toxicity or mortality were observed in any animal during the study period.

Under the test conditions, the test substance was considered to be non-irritating to 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:

From 19 October, 1992 to 09 November, 1992

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:

TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: Animal #1 and #2: Approx. 16 weeks; Animal #3: Approx. 13 weeks
- Weight at study initiation: 2499 to 3257 g
- Housing: Individually in labelled cages with perforated floors and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
- Diet (e.g. ad libitum): Standard laboratory rabbit diet (lKK-20, pellet diameter 4 mm, Hope Farms, Woerden, The Netherlands) approximately 100 g/d, ad libitum.
- Water (e.g. ad libitum): Tap-water diluted with decalcified water, ad libitum.
- Acclimation period: At least 5 d

ENVIRONMENTAL CONDITIONS
- Temperature: 21 °C
- Humidity: 55 %
- Air changes: Air-conditioned with 15 air changes/h
- Photoperiod: 12 h artificial fluorescent light/12 h dark

IN-LIFE DATES: From October 19, 1992 to November 9, 1992

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated eye

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (weight with unit): 100 ± 1 mg/animal

Duration of treatment / exposure:

Single exposure

Observation period (in vivo):

Up to 21 d (The eyes of each animal were examined for irritation for approx. 1, 24, 48 and 72 h and 7, 14 and 21 d after instillation of the test substance).

Number of animals or in vitro replicates:

Three male animals

Details on study design:

Prior to test substance instillation, the animals were physically examined to ensure normal ocular surface integrity.

SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: Fluorescein

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Slight blue discolouration of the cornea noted during 24, 48 and 72 h observation period.

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

fully reversible within: 21 d

Remarks on result:

other: The conjunctival redness was difficult to score at 24, 48 and 72 h observation period, due to blue discolouration by the test substance.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 7 d

Remarks on result:

other: Chemosis grade 2 (at 48 h) and grade 1 (at 72 h) for nictitating membrane.

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Slight blue discolouration of the cornea noted during 24, 48 and 72 h observation period.

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

24/48/72 h

Score:

2.33

Max. score:

3

Reversibility:

fully reversible within: 21 d

Remarks on result:

other: The conjunctival redness was difficult to score at 24 and 48 observation period, due to blue discolouration by the test substance. Redness grade 2 (at 24 and 48 h) and grade 3 (at 72 h) for nictitating membrane.

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 d

Remarks on result:

other: Chemosis grade 1 (at 48 h) for scleral conjunctivae.

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Slight blue discolouration of the cornea noted during 24, 48 and 72 h observation period.

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Time point:

24/48/72 h

Score:

2.67

Max. score:

3

Reversibility:

fully reversible within: 21 d

Remarks on result:

other: The conjunctival redness was difficult to score at 24 h observation period, due to blue discolouration by the test substance. Redness grade 2 (at 24 h) and grade 3 (at 48 and 72 h) for nictitating membrane.

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 d

Remarks on result:

other: chemosis grade 1 (at 48 h) for nictitating membrane.

Irritant / corrosive response data:

Blue staining of the cornea, iris and/or conjunctivae by the test substance was noted during the study, which made scoring difficult or impossible. Ocular corrosion was not observed in any of the animals.

Other effects:

There were no signs of body weight variability, toxicity or mortality in any animal during the study period.

None

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

The test substance was irritating to the eye.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance (of 100 % purity) in New Zealand White rabbits according to OECD Guideline 405 and EU Method B.5 in compliance with GLP.

A single dose of 100 ± 1 mg test substance was applied into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The untreated eye served as control. Without washing, eyes were scored for irritation reactions at 1, 24, 48 and 72 h and 7, 14 and 21 d post-instillation.

Blue staining of the cornea, iris and/or conjunctivae by the test substance at 24, 48 and 72 h made scoring difficult. During the study, blue staining of the fur on the head and paws was noted. Further, conjunctival redness of > 2 in 2/3 eyes was observed which had resolved within 21 d after instillation. There were no signs of body weight variability, toxicity or mortality in any animal during the study period.

Under the test conditions, the test substance was considered to be irritating to the eyes.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation

An in vivo study was performed to evaluate skin irritation/corrosion and eye irritation potency of the test substance following OECD guidelines and EU Method B.5 in compliance with GLP. Study performed in1992 showed very slight erythema in all three animals. The erythema had resolved within 24 h in all animals. Edema was not observed in any of the animals during the observation period. Blue staining of the treated skin by the test substance was observed in the three animals during the study. The body weights and the body weight gain of the animals were considered to be normal. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. Under the study conditions, the test substance was considered to be non-irritating to skin. Other two supporting studies performed in 1983, recorded data which is found to be non-irritating when applied to the rabbit skin.

Hence, based on the available data, Reactive Black 039 weas found to be non-irritant and non-corrosive to skin.

Eye Irritation

A study was conducted to assess the eye irritancy potential of the test substance (of 100 % purity) in New Zealand White rabbits according to OECD Guideline 405 and EU Method B.5 in compliance with GLP. A single dose of 100 ± 1 mg test substance was applied into the lower everted lid of one eye of each animal. The eyelids were then gently held together for one second before releasing. The untreated eye served as control. Without washing, eyes were scored for irritation reactions at 1, 24, 48 and 72 h and 7, 14 and 21 d post-instillation. Blue staining of the cornea, iris and/or conjunctivae by the test substance at 24, 48 and 72 h made scoring difficult. During the study, blue staining of the fur on the head and paws was noted. Further, conjunctival redness of > 2 in 2/3 eyes was observed which had resolved within 21 d after instillation. There were no signs of body weight variability, toxicity or mortality in any animal during the study period. Under the test conditions, the test substance was considered to be irritating to the eyes. 2 additional studies performed in 1983 are available to assess the eye irritancy potential of the test substance in New Zealand White rabbits with OECD guidelines 405. Both studies indicate the substance to be non-irritating. A similar supporting study performed in 1988 following the OECD guideline 405 reported that FAT 40171/C was found to be non-irritant and non-corrosive in the albino rabbit. Based on the data from all the above studies, Reactive Black 039 is considered to be non-irritant and non–corrosive to the skin and irritant to the rabbit eye.

Justification for classification or non-classification

Based on the available data, Reactive Black 039 does not warrant classification for skin irritation potential, however it needs to be classified as Eye Irritant category 2 according to Regulation (EC) No. 1272/2008 (CLP) criteria.