Tetrasodium 4,4'-bis[[4-[bis(2-hydroxypropyl)amino]-6-[(4-sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-stilbene-2,2'-disulphonate

Endpoint summary

Description of key information

Not skin irritating.

Not eye irritating.

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 April 23 to 26, 1986.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Nottingham University, School of Agriculture, Sutton Bonington, Leicestershire, U.K.
- Age at study initiation: approx. 12 to 16 weeks old.
- Weight at study initiation: 2.38 - 2.45 kg.
- Housing: animals were individually housed in suspended metal cages.
- Diet: free access, Rabbit Diet (Special Diet Services Limited, witham, Essex, U.K).
- Water: free access, driking water.
- Acclimation period: minimum five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 19 - 21 °C
- Humidity: 50 - 55 %
- Air changes: approx. 15 changes per hour.
- Photoperiod: 12 hours light and 12 hours darkness.

Preparation of test site:

clipped

Vehicle:

water

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g of test material, moistened with 0.5 ml of distilled water.
- Application: gauze patches of 2.5 x2.5 cm laden with the test material.

Duration of treatment / exposure:

Four hours after application the corset and patches were removed.

Number of animals:

Three.

Details on study design:

TEST SITE
- Area of exposure: approximately twenty-four hours prior to the commencement of the test, each of a group of three rabbits was prepared by closely clipping the fur from the dorsal/flank areas using veterinary clippers. On the day of the test a suitable test site was selected on the back of each rabbit.
- Type of wrap if used: the patch was secured in position over the test material with two lengths of adhesive strapping (SLEEK) in the form of a cross. To prevent the animals from interfering with the patches the trunk of each rabbit was wrapped in an elasticated corset (TUBIGRIP).

REMOVAL OF TEST SUBSTANCE
- Washing: any residual test material was removed by gentle swabbing with cotton wool soaked in distilled water.
- Time after start of exposure: 4 hours.

SCORING SYSTEM
Skin reactions were assessed on removal and after 48 and 72 hours and 4 and 7 days.
Erythema and Eschar Formation
No erythema: 0
Very slight erythema (barely perceptible): 1
Well defined erythema: 2
Moderate to severe erythema: 3
Severe erythema (beef redness) to slight eschar formation (injuries in depth): 4
Maximum possible: 4

Oedema 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 area of exposure): 4
Maximum possible: 4

Irritation parameter:

erythema score

Basis:

mean

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Irritation parameter:

edema score

Basis:

mean

Time point:

24/48/72 h

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

Very slight erythema was noted at one treated skin site one hour after removal of the patches. No adverse skin reactions were noted at this treated skin site at subsequent observations. The other treated skin sites appeared normal throughout the study period.

Tables of results is not available.

Interpretation of results:

not irritating

Remarks:

Migrated information according to the CLP Regulation Criteria used for interpretation of results: EU

Conclusions:

Not irritating

Executive summary:

Method

The skin irritation potential was tested in New Zealand White rabbits. The procedure used is described in the OECD guideline 404.

Results

Very slight erythema was noted at one treated skin site one hour after removal of the patches. No adverse skin reactions were noted at this treated skin site at subsequent observations. The other treated skin sites appeared normal throughout the study period.

Conclusion

The test material was found to be non-irritant to rabbit skin according to the EEC labelling Regulations and requires no symbol or risk phrase. Considering that the criteria to classify a substance according to the CLP regulation (EC 1272/2008) are similar, it can be assumed that also according to the CLP Regulation the substance is not irritating.

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 May 1 to 4, 1986.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted testing guidelines and performed according to GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Nottingham University, School of Agriculture, Sutton Bonington, Leicestershire, U.K.
- Age at study initiation: approx. 12 to 16 weeks old.
- Weight at study initiation: 2.4 - 2.69 kg.
- Housing: animals were individually housed in suspended metal cages.
- Diet: free access, Rabbit Diet (Special Diet Services Limited, witham, Essex, U.K).
- Water: free access, driking water.
- Acclimation period: minimum five days.

ENVIRONMENTAL CONDITIONS
- Temperature: 18 - 23 °C
- Humidity: 50 - 60 %
- Air changes: approx. 15 changes per hour.
- Photoperiod: 12 hours light and 12 hours darkness.

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.1 ml (which was found to weight 94 mg).

Number of animals or in vitro replicates:

Three.

Details on study design:

CONTROL BEFORE STARING
Within 24 hours of commencement of the test both eyes of each rabbit provisionally selected, were examined for evidence of ocular irritation or defect using an ophthalmoscope. Animals showing evidence of ocular lesions were rejected and replaced.

APPLICATION
The test substance was placed into the right eye of each rabbit by gently pulling the lower lid away from the eyeball to form a cup into which the test material was dropped. The upper and lower eyelids were held together for about one second immediately after application to prevent loss of the test material from the eye. The left eye remained untreated and was used for control purposes.
Following treatment of the first rabbit one crop or local anaesthetic (Ophthaine) was instilled into both eyes of the remaining two rabbits approximately 1 - 2 minutes before dosing.

SCORING SYSTEM
The eye irritation was assessed at 24, 48, 72 hours and 4 and 7 days after treatment.

CORNEA:
Opacity and degree of density (are most dense taken for reading) .
No ulceration or opacity: 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre) 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 cornea, iris not discernible through the opacity: 4

IRIS:
A Values
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia, or injection, any 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

CONJUNCTIVAE:
Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Blood vessels normal: 0
Some blood vessels definitely hyperaemic (injected): 1
Diffuse, crimson colour, individual vessels not easily discernible: 2
Diffuse beefy red: 3

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

Irritation parameter:

cornea opacity score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Irritation parameter:

iris score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 1

Reversibility:

fully reversible

Irritation parameter:

conjunctivae score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible

Irritation parameter:

chemosis score

Basis:

animal: 3/3

Time point:

24/48/72 h

Score:

< 2

Reversibility:

fully reversible

Irritant / corrosive response data:

A dulling of the normal lustre of the corneal surface was noted in one treated eye one hour after treatment; no corneal opacity was noted in this eye at subsequent observations. The cornea of the other treated eyes appeared normal throughout the study period.
Iridial inflammation was noted in two treated eyes one hour after treatment. The iris of the remaining treated eye appeared normal at this time. No iridial inflammation inflammation was noted at the 24, 48 and 72-hour observations.
Conjunctivitis was noted in all treated eyes one and twenty-four hours after treatment. All treated eyes were normal at the 48 and 72-hour observations.

Animal	1 hr	24 hrs	48 hrs	72 hrs	Mean 24, 48, 72 hrs
Cornea
1 (6)	0	0	0	0	0.00
2 (64)	0	0	0	0	0.00
3 (71)	0	0	0	0	0.00
Iritis
1 (6)	1	0	0	0	0.00
2 (64)	1	0	0	0	0.00
3 (71)	0	0	0	0	0.00
Conjunctival - Redness
1 (6)	2	1	0	0	0.33
2 (64)	2	1	0	0	0.33
3 (71)	1	1	0	0	0.33
Conjunctival - Chemosis
1 (6)	1	0	0	0	0.00
2 (64)	1	0	0	0	0.00
3 (71)	1	0	0	0	0.00

Interpretation of results:

not irritating

Remarks:

Migrated information according to the CLP Regulation Criteria used for interpretation of results: EU

Conclusions:

Not irritating.

Executive summary:

Method

The test was performed on 3 New Zealand White rabbits. The test material in an amount of 0.1 g was inserted into placed into the right eye of the rabbits.

The eye irritation assessment was made at 1, 24, 48 and 72 hours.

Results

A dulling of the normal lustre of the corneal surface was noted in one treated eye one hour after treatment; no corneal opacity was noted in this eye at subsequent observations. The cornea of the other treated eyes appeared normal throughout the study period.

Iridial inflammation was noted in two treated eyes one hour after treatment. The iris of the remaining treated eye appeared normal at this time. No iridial inflammation inflammation was noted at the 24, 48 and 72-hour observations.

Conjunctivitis was noted in all treated eyes one and twenty-four hours after treatment. All treated eyes were normal at the 48 and 72-hour observations.

Conclusion

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for irtitis, lower than 2 for both conjunctival redness and oedema, in all animals.

Therefore the substance does not meet the criteria to be classified as eye irritating.

In conclusion, the test item can be classified as NON irritating, according to the the CLP (EC 1272/2008) Regulation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Valid GLP studies with New Zealand White rabbits on OB 4 -MSA were performed for skin and eye irritation with negative results (Jones and Collier, 1986).

The results on the most similar substance within the category of Stilbene Fluorescent Whitening Agents (OB 3a-MSA) are here reported as supporting data (Ciba-Geigy Ltd., 1981).

This substance can be considered the best representative within the category, since it has the same sulphonation degree than the substance under registration CAS 67786-25.8, a very high structural similarity (Tanimoto distance > 0.9), very similar molecular weight (1165 Vs 1121, respectively) and a very high water solubility (650 g/l Vs 1000 g/l, respectively).

Furthermore, the hydroxylic function in the substance under registration is a secondary one, with a lower reactivity and a higher steric hindrance respect to the analogue substance that has the primary hydroxylic function, more reactive. In fact no breakdown metabolisation products are proposed on both substances by the OECD metabolism skin simulator.

Within the whole category ten over fourteen registered substances were tested and none of the existing tests arisen any concern for skin and eye irritation.

All substances of the category were modelled with OECD Toolbox and the provisional results about skin and eye irritation were calculated for all members. No alerts were reported (see category Justification Report).

Justification for classification or non-classification

According to the CLP Regulation (EC 1272/2008), 3.2 Skin corrosion/irritation section, skin Irritation means the production of reversible damage to the skin following the application of a test substance for up to 4 hours.

In all the studies available, the mean values from gradings at 24, 48 and 72 hours after patch removal were lower than 2.3 in at least 2 of 3 animals for both erythema/eschar and oedema reactions.

According to the Regulation EC 1272/2008 (CLP) substances that have the potential to induce reversible eye irritation are classified in Category 2 (irritating to eyes) if, when applied to the eye of an animal, a substance produces at least in 2 of 3 tested animals, a positive response of corneal opacity ≥ 1 and/or iritis ≥ 1, and/or conjunctival redness ≥ 2 and/or conjunctival oedema (chemosis) ≥ 2, calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days.

The mean values from gradings at 24, 48 and 72 hours were lower than 1 for corneal opacity, lower than 1 for irtitis, lower than 2 for both conjunctival redness and oedema, in all animals. Therefore the substance does not meet the criteria to be classified as eye irritating.

The available experimental data are adequate for classification and labelling. The substance is not classified for eye and skin irritation, according to the CLP Regulation (EC 1272/2008).