Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October 1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with GLP, however no test guidance is referenced in the study report. The methodology is equivalent to OECD Guideline 405 (Acute Eye Irritation / Corrosion).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Principles of method if other than guideline:

No test guideline referenced within the study report. The methodology is equivalent to OECD Guideline 405 (Acute Eye Irritation / Corrosion).

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

EC Number:

413-550-5

EC Name:

Octasodium 2-(8-(4-chloro-6-(3-((4-chloro-6-(3,6-disulfonato-2-(1,5-disulfonatonaphthalen-2-ylazo)-1-hydroxynaphthalen-8-ylamino)-1,3,5-triazin-2-yl)aminomethyl)phenylamino)-1,3,5-triazin-2-ylamino)-3,6-disulfonato-1-hydroxynaphthalen-2-ylazo)naphthalene-1,5-disulfonate

Cas Number:

142068-96-0

Molecular formula:

Hill formula: C53H28Cl2N14Na8O26S8

IUPAC Name:

octasodium 5-[(4-chloro-6-{[4-({[4-chloro-6-({7-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-8-hydroxy-3,6-disulfonatonaphthalen-1-yl}amino)-1,3,5-triazin-2-yl]amino}methyl)phenyl]amino}-1,3,5-triazin-2-yl)amino]-3-[2-(1,5-disulfonatonaphthalen-2-yl)diazen-1-yl]-4-hydroxynaphthalene-2,7-disulfonate

Details on test material:

The test sample (reference: NBY 405/74) was received as a dark brown powder. It was sent for testing from ICI Specialties, Blackley, Manchester, UK. A certificate of analysis (reference: 9108127, dated 8 August 1991) stated that the test sample contained 71.6% w/w Substance H112339, From information supplied by the sponsor the test sample was used within the stated expiry date. . The test sample was given the CTL reference number Y07719/001 and was tested as supplied.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Mellor Rabbits, UK
- Age at study initiation: Young adult
- Weight at study initiation: 3371-4417g
- Housing: The rabbits were housed individually in anodised aluminium sheet cages (65,2cm width x 68.9cm depth x 45.7cm height). The floor of each cage was made of perforated aluminium sheet and the door was made of stainless steel rods.
- Diet (e.g. ad libitum): Labsure CRB Rabbit Diet, ad libitum
- Water (e.g. ad libitum): via an automatic system, ad libitum
- Acclimation period: minmum of 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17°C (± 2°C)
- Humidity (%): 55 (±15%)
- Air changes (per hr): 25-30
- Photoperiod (hrs dark / hrs light): controlled by a time switch to give alternate periods of 12 hours of artificial light and 12 hours of darkness.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100mg as supplied.

Duration of treatment / exposure:

7 days

Observation period (in vivo):

None.

Number of animals or in vitro replicates:

3

Details on study design:

Both eyes of each rabbit were examined within twenty-four hours prior to the study. The examination consisted of a visual assessment with the aid of fluorescein and only rabbits without any apparent eye defects or ocular irritation were used.

Three female rabbits were used for this study. Each animal was given a number, unique within the study, which was written on the inner side of one ear, using a black waterproof marker-pen, and displayed on the cage card. The bodyweight of each animal was recorded at the start of the study.

Initially the test sample (approximately l00mg) was applied into the conjunctival sac of the left eye of one rabbit by gently pulling the lower lid away from the eyeball to form a cup into which the test sample was dropped. The lids were then gently held together for 1-2 seconds after which the animal was released. The other eye was untreated (control eye).

Immediately after the application of the test sample, an assessment of the initial pain reaction of the rabbit was made using a six-point scale.

Eight days later, the test sample was applied into the test eye of the remaining two animals as described previously and an assessment of initial pain reaction was made.

The eyes were examined and the Draize scale (Draize 1959) was used to assess the grade of ocular reaction at intervals. In addition, as an aid in the assessment of corneal damage, fluorescein staining was used at the 1, 2, 3, 4, 6 and 7-day readings . A modified form of the Kay and Calandra system (1969) was used to interpret and classify the numerical scores.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: Mean 0-7 days

Score:

> 1 - < 2

Max. score:

4

Reversibility:

fully reversible within: 2 days

Results 2

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: Mean 0-7 days

Score:

> 1 - < 2

Max. score:

4

Reversibility:

fully reversible within: 4 days

Results 3

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: Mean 0-7 days

Score:

> 1 - < 2

Max. score:

4

Reversibility:

fully reversible within: 4 days

Irritant / corrosive response data:

Application of the test sample into the conjunctival sac of three rabbits caused slight initial pain.

The test sample stained the tissues of the eye red and prevented a complete assessment of ocular irritation. Corneal and iridial effects were completely obscured in one animal throughout the study and in the remaining two animals up to day 3 of the study. From day 4 after application there were no corneal or iridial effects in these two animals.

Due to the red staining, it was not possible to assess conjunctival redness in any animal during the study. However, slight to mild chemosis and slight to severe discharge were observed in all animals up to day 3 of the study. Additional signs of ocular irritation included dried secretions of the periorbital skin in 1 animal and slight to moderate Harderian discharge in 2 of the rabbits. Following instillation of fluorescein stain, up to
one quarter of the cornea of one rabbit became stained, indicating possible corneal damage. This animal showed no reaction to fluorescein stain from day four of the study. None of these additional signs of ocular irritation were evident in any animal from day 4 after application of the test sample.

Applicant's summary and conclusion

Interpretation of results:

slightly irritating

Remarks:

Migrated information however due to staining of the eye, a higher classification is applicable. Criteria used for interpretation of results: EU

Conclusions:

Application of the test sample into the conjunctival sac of three rabbits caused slight initial pain (class 2 on a 0-5 scale). On the basis of the available scores, Substance H112339 was at least a mild irritant (class 4 on a 1-8 scale) to the rabbit eye. However, the test sample is unlikely to be a severe ocular irritant since no signs of irritation were evident from day 4 after dosing. However, due toe staining effects noted in the eye, a higher classification is applicable.

Executive summary:

Study conducted in compliance with GLP, however no test guidance is referenced within the study report.

On the basis of the available scores, the test sample is a mild irritant to the rabbit eye, however it is unlikely to be a severe ocular irritant since no signs of irritation were evident from 4 days after dosing. 

The test material also caused staining of the cornea; however this was not noted after 4 days in 2 rabbits. Staining was noted in one rabbit only on day 7, and receding effects indicate that this would not be present at the 21-day observation. However, review of REGULATION (EC) No 1272/2008 on classification, labelling and packaging of substances and mixtures, specificallysection3.3.2.6.Irreversible effects on the eye/serious damage to eyes (Category 1) states that:

 

3.3.2.6.1. Substances that have the potential to seriously damage the eyes are classified in Category 1 (irreversible effects on the eye). Substances are classified in this hazard category on the basis of the results of animal testing, in accordance with the criteria listed in Table 3.3.1. 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.

 

As a dye substance, the test material was not considered to cause discoloration of the cornea at the 21-day observation point as specified in the regulation, and hence is not considered to fulfil the criteria for classification as a Category 1 eye irritant. No classification is therefore required.