reaction mass of: 7-amino-3,8-bis-[4-(2-sulfoxyethylsulfonyl)phenylazo]-4- hydroxynaphthalene-2-sulfonic acid, Na/K salt; 7-amino-3-[4-(2-sulfoxyethylsulfonyl)phenylazo]-4-hydroxy-8-[4-(2-sulfoxyethylsulfonyl)-2- sulfophenylazo]naphthalene-2-sulfonic acid, Na/K salt; 7-amino-8-[4-(2-sulfoxyethylsulfonyl)-phenylazo]-4-hydroxy-3-[4-(2-sulfoxyethylsulfonyl)- 2-sulfophenylazo]naphthalene-2-sulfonic acid, Na/K salt; 7-amino-3,8-bis-[4-(2-sulfoxyethylsulfonyl)-2-sulfophenylazo]-4-hydroxynaphthalene-2- sulfonic acid, Na/K salt

Endpoint summary

Description of key information

No irritating effects were seen in the skin and eye irritation studies. However, staining of the different tissues in the eye was observed.

Staining of cornea and iris were no longer present after 7 days, but scleral and conjunctival (nictitating membrane) staining was not completely reversible after 21 days.

As the irreversible staining does not impair sight, it is not relevant for classification and labelling according to the CLP guideline.

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 6, 1998 to 03 Nov 1998

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:

not specified

Qualifier:

according to guideline

Guideline:

other: Directive 92/69 EEC, B.4. "Acute Toxicity - Skin Irritation", July 31, 1992.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: TVR50
- Expiration date of the lot/batch: September 1, 2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature (ca. 20°C) away from direct sunlight.
- Stability under test conditions: Stable at storage conditions.
- Stability of the test substance in the vehicle: Stable for 24 h in bidistilled water

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne
- Age at study initiation: 15 weeks
- Housing: Individually in stainless steel cages with an automatic cleaning system equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet: Pelleted standard Provimi Kliba 3410 rabbit maintenance diet ad libitum (batch no. 62/98). Results of analysis are archived by RCC.
- Water: Community tap water from Itingen, ad libitum, in water bowls. Results of analysis are archived by RCC.
- Acclimation period: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ±3
- Humidity (%): 40 - 70
- Air changes (per hr): 10 - 15 air changes per h
- Photoperiod (hrs dark / hrs light): by fluorescent light on a 12 h light/dark cycle.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

water

Remarks:

bidistilled water

Controls:

other: opposite flank

Amount / concentration applied:

500 mg

Duration of treatment / exposure:

4 h

Observation period:

24, 48 and 72 h

Number of animals:

1 male
2 females

Irritation parameter:

erythema score

Basis:

mean

Remarks:

3 rabbits

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Remarks:

3 rabbit

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

IRRITATION : No signs of irritation were observed.

CORROSION : No irreversible alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

COLORATION : Red staining by the test article of the treated skin was observed in all animals up to the end of the observation period.
VIABILITY/MORTALITY AND CLINICAL SIGNS :
No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 40'571/A is considered to be not irritating to rabbit skin.

Executive summary:

Primary skin irritation study with FAT 40571/A was carried out in New Zealand White according to OECD 404 and EU B 4 guideline with semi-occlusive type of coverage. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 h after removal of the dressing. The scores of each animal at the reading times 24, 48, 72 h were used in calculating the respective mean values for each type of lesion.

The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 h and then dividing by the number of data points. The primary irritation score was 0.00 (max. 8.0).

Local signs (mean values from 24 to 72 h) consisted of grade 0.00 erythema and grade 0.00 edema. No signs of irritation were observed. The test article caused red staining of the treated skin.

No corrosive effects were noted on the treated skin of any animal at any measuring interval.

Based on the study results the test article FAT 40571/A is considered to be "not 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:

Oct 13, 1998 to Nov 27,1998

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:

not specified

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: TVR50
- Expiration date of the lot/batch: September 1,2004

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature (ca. 20 °C) away from direct
sunlight
- Stability under test conditions: Stable at storage conditions

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes, F-01400 Chatillon sur Chalaronne
- Age at study initiation: 15 weeks
- Weight at study initiation: male : 3069 g ; females : 3308 - 3190 g
- Housing: Individually in stainless steel cages with an automatic cleaning system equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet: Pelleted standard Provimi Kliba 3410 rabbit maintenance diet ad libitum (batch no. 62/98). Results of analysis are archived by RCC.
- Water: Community tap water from Itingen, ad libitum, in water bowls. Results of analysis are archived by RCC.
- Acclimation period: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ±3
- Humidity (%): 40 - 70
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): fluorescent light on a 12 h light/dark cycle

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.1 g per animal

Duration of treatment / exposure:

The test article (undiluted) was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test article.

Observation period (in vivo):

Viability/mortality : Daily.
Clinical signs : Daily during the observation period.
Body weights : At start of acclimatization, on the first day of application and at termination of observation.
Scoring of irritation : performed approximately 1, 24, 48 and 72 h as well as 7, 14 and 21 days after test article application. The scores of each animal at the following reading times (24, 48 and 72 h) were used in calculating the respective mean values for each type of lesion.

Number of animals or in vitro replicates:

3 (1 male, 2 females)

Details on study design:

The eyes of the animals were examined one day prior to test article administration. Animals with overt signs of ocular injury or irritation which may have interfered with the interpretation of the results were not used in the test.
On the day of treatment, the test article (undiluted) was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of test article. The right eye remained untreated and served as the reference control. In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes (Washington, 1977).

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritation parameter:

iris score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.66

Max. score:

3

Reversibility:

fully reversible within: 14 days

Remarks on result:

probability of weak irritation

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 14 days

Remarks on result:

probability of weak irritation

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

no indication of irritation

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animal

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Effects on redness of eyes were reversible within 14 days but difficulties were seen during the first three days to score the eyes due to the staining caused by the substance.
No corrosion of the cornea was observed at any of the reading times.

Other effects:

COLORATION: Reversible orange staining of the cornea, iris, conjunctivae (including nictitating membrane) and lid hairs by the test article was observed.
Scleral and conjunctival (nictitating membrane) staining was not reversible in all animals after 21 days.

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.

 

IRRITATION

Application of the test article to healthy rabbit conjunctivae resulted in a primary irritation score of 0.33. Slight swelling of the conjunctivae including nictitating membrane and slight watery discharge was observed in all animals one hour after treatment. Slight conjunctival reddening was noted until day 7 of the observation period.

 

COLORATION

Orange staining of the cornea, iris, conjunctivae (including nictitating membrane), lid hairs and sclera by the test article was observed. Scleral and conjunctival (nictitating membrane) staining was not reversible in all animals after 21 days.

 

BODY WEIGHTS

The body weight of all rabbits were considered to be within the normal range of variability.

 

Individual eye scores

Animal Number	Time point	Cornea	Iris	Conjunctiva
				Redness	Chemosis
22	24 h	0	0	*	0
23		0	0	*	0
24		0	0	*	0
22	48 h	0	0	1	0
23		0	0	*	0
24		0	0	*	0
22	72 h	0	0	1	0
23		0	0	1	0
24		0	0	*	0

 

*      : Reaction and/or staining hampering evaluation

Interpretation of results:

GHS criteria not met

Conclusions:

The substance is classified as not irritating to eyes. As the irreversible staining does not impair sight, it is not relevant for classification and labeling according to the CLP guideline

Executive summary:

The primary irritation potential of FAT 40'571/A was investigated in New Zealand White rabbits according to OECD 405 and B.5 guideline by instillation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. The treated eyes were not rinsed after application. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 h as well as 7, 14 and 21 days after test article application. The scores of each animal at the following reading times (24, 48 and 72 h) were used in calculating the respective mean values for each type of lesion.

Eye irritation scores found in animal number 22 after 48 h, animal 23 after 72 h and 7 days. These effects were subsided after 14 days. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 h and then dividing the resulting total by the number of data points. The primary irritation score was 0.33 (max. 13). Swelling of the conjunctivae including nictitating membrane and watery discharge was observed in all animals one hour after treatment. Slight conjunctival reddening was noted until day 7 of the observation period. Orange staining of the cornea, iris, conjunctivae (including nictitating membrane), lid hairs and sclera of the treated eyes by the test article was observed. Scleral and conjunctival (nictitating membrane) staining was not reversible after 21 days, all other effects were reversed by Day 7. No corrosion was observed at any of the measuring intervals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin Irritation:

Primary skin irritation study with Reactive Brown 49 was carried out in New Zealand White according to OECD 404 and EU B 4 guideline with semi-occlusive type of coverage. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 h after removal of the dressing. The scores of each animal at the reading times 24, 48, 72 h were used in calculating the respective mean values for each type of lesion.

The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 h and then dividing by the number of data points. The primary irritation score was 0.00 (max. 8.0).

Local signs (mean values from 24 to 72 h) consisted of grade 0.00 erythema and grade 0.00 edema. No signs of irritation were observed. The test article caused red staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any measuring interval.

 

Eye irritation

The primary irritation potential of Reactive Brown 49 was investigated in New Zealand White rabbits according to OECD 405 and B.5 guideline by instillation of 0.1 g into one eye of each of three young adult New Zealand White rabbits. The treated eyes were not rinsed after application. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 h as well as 7, 14 and 21 days after test article application. The scores of each animal at the following reading times (24, 48 and 72 h) were used in calculating the respective mean values for each type of lesion.

Eye irritation scores found in animal number 22 after 48 h, animal 23 after 72 h and 7 days. These effects were subsided after 14 days. The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 h and then dividing the resulting total by the number of data points. The primary irritation score was 0.33 (max. 13). Swelling of the conjunctivae including nictitating membrane and watery discharge was observed in all animals one hour after treatment. Slight conjunctival reddening was noted until day 7 of the observation period. Orange staining of the cornea, iris, conjunctivae (including nictitating membrane), lid hairs and sclera of the treated eyes by the test article was observed. Scleral and conjunctival (nictitating membrane) staining was not reversible after 21 days, all other effects were reversed by Day 7. No corrosion was observed at any of the measuring intervals.

Justification for classification or non-classification

Based on the findings in the skin and eye irritation study the Reactive Brown 49 is not considered to be classified according to the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.

The substance is classified as H318 in Annex VI: Index Number: 611-144-00-4; due to irreversible staining of the eye. However, according to the new CLP regulation, classification due to irreversible staining is only relevant if vision is impaired. As only the conjunctiva and sclera is affected, the substance no longer classifies as eye irritant.