sodium 5-(4-amino-6-(4-((E)-4-(4-(bis(2-hydroxyethyl)amino)-6-(8-hydroxy-7-((E)-phenyldiazenyl)-3,6-disulfonatonaphthalen-1-ylamino)-1,3,5-triazin-2-ylamino)-2-sulfonatostyryl)-3-sulfonatophenylamino)-1,3,5-triazin-2-ylamino)-4-hydroxy-3-((E)-phenyldiazenyl)naphthalene-2,7-disulfonate

Endpoint summary

Description of key information

Not skin irritant

Not eye irritant

Key value for chemical safety assessment

Skin irritation / corrosion

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

No information on "irritation/corrosion" of the Target Substance is available. However data on a similar substance (Similar Substance 01) has been taken into account for the assessment. More details on the substance similarity are reported in section 13.

Skin Irritation/Corrosion

Key study

A skin irritation study in three New Zealand White rabbits (2 males, 1 female) was performed according to OECD guideline 404 and EU Method B.4. Therefore, 0.5 g of the test substance (60 % act. ingr.) was applied to approx. 6 square cm of the intact skin of the clipped area and covered in a semi-occlusive manner for 4 hours. The skin reaction was assessed at 1, 24, 48 and 72 hours after the removal of the test substance. Local signs (mean values from 24, 48, 72 hours) consisted of grade 0.0 erythema and grade 0.0 edema. No acute clinical symptoms were observed in the animals during the test and observation period, and no mortality occurred. In the area of application a red staining of the treated skin by pigment or coloring of the test article was observed.

Supporting study

An acute dermal irritation study was performed in accordance to OECD guideline No. 404 and Method B4 in Commission Directive 92/69/EEC to determine the irritancy potential of the test material (49 % act. ingr.) to the skin of three New Zealand White rabbits (2 females, 1 male). The rabbits were clipped free of fur from the dorsal flank area using veterinary clippers 24 hours before treatment. On the day of the test, a quantity of 0.5g of the test material, moistened with 0.5 mL of distilled water, was introduced under a 2.5 cm x 2.5 cm cottongauze patch and placed in position on the shorn skin. The patch was secured in position with a strip of surgical adhesive tape (BLENDERM: 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) and the animals were returned to their cages for the duration of the exposure period. 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 74% Industrial Methylated Spirits. 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 following scale from Draize. Pink staining of the treatment site was noted in one animal at the 1, 24, 48 and 72-hour observations. Very slight erythema was noted at all treatment sites at the 1-hour observation and two treatment sites at the 24, 48 and 72-hour observations. Desquamation was noted at two treatment sites at the 7-day observation. An isolated incident of very slight oedema was noted at one treatment site at the 24-hour observation.

Both the key and supporting study indicated that the test material was not irritating to the rabbits' skin.

Eye Irritation/Corrosion

Key study

An eye irritation study in three New Zealand rabbits (1 male, 2 females) was performed according to OECD guideline 405 and EU Method B.5. 0.1 g of the test substance (60 % act. integr.) was placed in the left eye of each animal and the lids were gently held together for about one second. The eyes of each animal were exmamined for ocula irritation 1, 24, 48, 72 hours, 7, 14 and 21 days after administration.

No necropsy was performed in the animals euthanized at termination of observation. No acute clinical symptoms were observed in the animals during the test and observation period, and no mortality occured. The body weight gain of all rabbits was similar. The mean scores (over time points 24 h, 48 h, 72 h) for animal 1 were determined to be 0.0 for cornea, 0.0 for iris, 1.0 for conjunctivae redness and 0.0 for chemosis. For animal 2, the mean scores were 0.33 for cornea, 0.0 for iris, 1.0 for conjunctiva redness and 0.33 for chemosis. For animal 3, the mean scores were 0.0 for cornea, 0.0 for iris, 1.0 for conjunctiva redness and 1.0 for chemosis. The observed effects were fully reversible within the 21 -day observation period.

Supporting study

An acute eye irritation study was performed in accordance to OECD guideline No. 405 and Method B5 in Commission Directive 92/69/EECto determine the irritancy potential of the test material (49 % act. integr.) to the eyes of three New Zealnd White rabbits (2 females, 1 male). A volume of 0.1 mL of the test material, which was found to weigh approximately 98 mg was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after application, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes.In order to minimise pain on instillation of the test material, one drop of local anaesthetic ("Ophthaine", 0.5% proxymetacaine hydrochloride, E.R. Squibb & Sons Limited, Hounslow, Middlesex, U.K.) was instilled into both eyes of these animals 1 - 2 minutes before treatment. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment in accordance to the OECD scoring system. Examination of the eye was facilitated by use of the light source from a standard ophthalmoscope. Additional observations were made on days seven, fourteen and twenty-one to assess the reversibility of the ocular effects. Residual test material was noted in all treated eyes throughout the study period. The mean scores (over time points 24 h, 48 h, 72 h) for animal 1 were determined to be 0 for cornea, 0 - 1.3 for iris, 0.7 - 1.7 for conjunctivae redness and 1.3 for chemosis. Dark pink/purple-coloured staining prevented accurate evaluation of iridial inflammation in the treated eye of animal 2 at the 24 and 48 -hour observations. For animal 2, the mean scores were 0 for cornea, 0 for iris, 1 for conjunctiva redness and 0.7 for chemosis. For animal 3, the mean scores were 1.0 for cornea, 0.3 for iris, 1.7 for conjunctiva redness and 1.3 for chemosis. Likewise, vascularisation in animal 3 was observed at the 7 and 14 -day obervations. The observed irritation paramters were fully reversible within the observation period.

On the basis of the key and supporting study, the test material is considered to be non-irritating to the rabbits' eye.

Justification for selection of skin irritation / corrosion endpoint:

OECD guideline study; test material with highest concentration of active ingredient

Justification for selection of eye irritation endpoint:

OECD guideline study; test material with highest concentration of active ingredient

Justification for classification or non-classification

SKIN IRRITATION/CORROSION

A corrosive substance is a substance that produces destruction of skin tissue, namely, visible necrosis through the epidermis and into the dermis, in at least 1 tested animal after exposure up to 4 hour duration.

Three subcategories are provided within the corrosive category 1:

Subcategory 1A where responses are noted following up to 3 minutes exposure and up to 1 hour observation;

Subcategory 1B where responses are described following exposure between 3 minutes and 1 hour and observations up to 14 days;

Subcategory 1C where responses occur after exposures between 1 hour and 4 hours and observations up to 14 days

To classify a substance as irritant, the major criterion is that at least 2 of 3 tested animals have a mean score of ≥ 2,3 - ≤ 4,0.

Category 2:

- Mean value of ≥ 2,3 - ≤ 4,0 for erythema/ eschar or for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal or, if reactions are delayed, from grades on 3 consecutive days after the onset of skin reactions; or

- Inflammation that persists to the end of the observation period normally 14 days in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling; or

- In some cases where there is pronounced variability of response among animals, with very definite positive effects related to chemical eposure in a single animal but less than the criteria above.

 

EYE IRRITATION

Serious eye damage means the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance to the anterior surface of the eye, which is not fully reversible within 21 days of application.

Eye irritation means the production of changes in the eye following the application of test substance to the anterior surface of the eye, which are fully reversible within 21 days of application.

Irreversible effects on the eye (Category 1):

If, when applied to the eye of an animal, a substance produces:

- at least in one animal effects on the cornea, iris or conjunctiva that are not expected to reverse or have not fully reversed within an observation period of normally 21 days; and/ or

- at least in 2 of 3 tested animals, a positive response of corneal opacity ≥ 3 and/or iritis > 1,5 calculated as the mean scores following grading at 24, 48 and 72 hours after installation of the test material.

These observations include animals with grade 4 cornea lesions and other severe reactions (e.g., destruction of cornea) observed at any time during the test, as well as persistent corneal opacity, discoloration of the cornea by a dye substance, adhesion, pannus, and interference with the function of the iris or other effects that impair sight. In this context, persistent lesions are considered those which are not fully reversible within an observation period of normally 21 days. Substances are also classified in Category 1 if they fulfil the criteria of corneal opacity ≥ 3 or iritis > 1,5 detected in a Draize eye test with rabbits, recognising that such severe lesions usually do not reverse within a 21-day observation period.

The substance does not meet the requirements to be classified in Category 1.

Irritating to eyes (Category 2)

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.

Based on the results obtained in the skin and eye irritation studies, the test substance does not need to be classified for skin and eye irritation according to the CLP Regulation (EC) No. 1272/2008.