Pentasodium 7-(4-(4-(5-amino-4-sulfonato-2-(4-((2-(sulfonato-ethoxy)sulfonyl)phenylazo)phenylamino)-6-chloro-1,3,5-triazin-2-yl)amino-2-ureidophenylazo)naphtalene-1,3,6-trisulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 November 1995 to 25 December 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OTS 798.4500 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

FAT 40544/A

IUPAC Name:

FAT 40544/A

Test material form:

solid: compact

Details on test material:

Identification: CIBACRON GELB TZ 4210 ROH LÖSUNG (LABORGETROCKNET) (FAT 40'544/A)
Remark: FAT 40544 and FAT 40549 are chemically identical. FAT 40544 is typically containing more moisture, which has been removed for this experiment, thus making the sample representative for FAT 40549 too.
Description: red powder
Batch number: Vers. Nr. TV 1
Purity/formulation: approx. 90%
pH (1 g/l): 5 - 6
Stability of test article: Stable at storage conditions; expiration date: November 2000
Storage conditions: At room temperature (approx. 20 °C), away from direct sunlight
Safety precautions: Gloves, goggles and face mask were sufficient to ensure personnel health and safety.

Specific details on test material used for the study:

Identification: CIBACRON GELB TZ 4210 ROH LÖSUNG (LABORGETROCKNET) (FAT 40'544/A)
Description: Red powder
Batch number: TV.1
Purity/formulation: ca 90%
pH (1 g/L): 5-6
Stability of test article: stable at storage conditions: expiration date: Nov 2000
Storage conditions: At room temperature (ca 20 °C) away from sunlight.
Safety precautions: Gloves, goggles and face mask were sufficient to ensure personnel health and safety.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test animals: New Zealand White Rabbit - CRL:KBL(NZW)BR
Animal Source: Charles River, Stolzenseeweg 32-36, D-88353 Kisslegg
Age at treatment: 13 weeks
Number of animals: 3 (1 male and 2 females)
Identification: By unique cage number and corresponding ear number.
Acclimatization: Four days under laboratory conditions after health examination. Only healthy animals were used for the study.
Body weight at start of acclimatization: male : 2.2 kg / females: 2.2 - 2.3 kg
Body weight at start of treatment: male: 2.4 kg / females: 2.3 - 2.5 kg
Standard Laboratory Conditions: Air-conditioned with 10 - 15 air changes per hour and continuously monitored environment with target ranges for temperature of 20 ±3 °C and for relative humidity between 40 - 70 % (values above 70% during cleaning process possible), 12 hours artificial fluorescent light/12 hours dark, music during the light period.
Accommodation: Individually in stainless steel cages with an automatic cleaning system, equipped with feed hoppers, drinking water bowls and wood for gnawing.
Diet: Pelleted standard Kliba 341, Batch no. 92/95 rabbit maintenance diet (Kliba Mühlen AG, CH-4303 Kaiseraugst), ad libitum. Results of analysis for contaminants are included in the report.
Water: Community tap water from Itingen, ad libitum.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g of 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.

Duration of treatment / exposure:

On test day 1, 0.1 g of 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 (eyes remained unrinsed). The lids were then gently held together for about one second to prevent loss of the test article. The right eye remained untreated and served as the reference control.

Observation period (in vivo):

Viability/Mortality and Clinical Signs were checked daily during the observation period. Body weights were recorded at start of acclimatisation, at start of treatment and following treatment. The eyes of each animal were examined for ocular irritation approximately 1, 24, 48 and 72 hours, as well as 7, 14 and 21 days after administration.

Number of animals or in vitro replicates:

3 (1 male and 2 females)

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: No

EVALUATION OF EYE REACTIONS
CORNEAL IRRITATION
Opacity: degree of density (densest area used for assessment)
No ulceration or opacity - 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details of iris clearly visible - 1
Easily discernible translucent area, details of iris slightly obscured - 2
Nacrous area, no details of iris visible, size of pupil barely discernible - 3
Opaque cornea, iris not discernible through the opacity - 4

IRIDIC IRRITATION
Normal - 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any of these or combination thereof, iris still reacting to light (sluggish re action is positive) - 1
No reaction to light, hemorrhage, gross destruction (any or all of these) - 2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebral and bulbar conjunctivae, excluding cornea and iris).
Blood vessels normal - 0
Some blood vessels definitely hyperemic (injected) - 1
Diffuse, crimson color, individual vessels not easily discernible - 2
Diffuse beefy red - 3

Chemosis: lids and/or nictitating membranes
No swelling - 0
Any swelling above normal (includes nictitating membranes) - 1
Obvious swelling with partial eversion of lids - 2
Swelling with lids about half closed - 3
Swelling with lids more than half closed - 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.78

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

CIBACRON GELB TZ 4210 ROH LÖSUNG (LABORGETROCKNET) (FAT 40'544/A) showed a primary irritation score of: 0.78, when applied to the conjunctival sac of the rabbit eye.
One (1) hour after instillation of the test article, moderate reddening and swelling of the conjunctivae were noted in all animals, as was moderate to marked watery discharge. Hyperemia of the scleral blood vessels was also evident in all animals.
Reddening and swelling of the conjunctivae tissues were reversible after 72 hours in two animals and after seven days in one animal.

Other effects:

Light orange to orange-brown staining of the conjunctivae was noted in all animals until observation on day 14. From day 14 onwards, staining of the conjunctivae was yellow. Staining of the sclera was evident in all animals on day 21 after application. The corneas of all treated animals were unaffected.

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred. No corrosion of the cornea was observed at any of the reading times. The body weight of the animals was within the normal range of variability.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance is considered non-irritant to rabbit eyes.

Executive summary:

The primary irritation potential of the test article was investigated according to method specified as per OECD test guideline 405 in a GLP certified laboratory.

0.1 g of test substance was instilled into conjuctival sac of each of three adult New Zealand rabbits. The treated eyes were not rinsed after application. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours 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 hours) were used in calculating the respective mean values for each type of lesion.

The primary irritation score was calculated by averaging the individual cumulative scores at 24, 48 and 72 hours. Corneal opacity, iris and chemosis scores were zero at 24, 48 and 72h. Conjuctivae redness score was 0.78 which was reversible in 7 days.

Hyperemia of the scleral blood vessels was evident in all animals. No finding persisted longer than seven days. Light orange to orange-brown staining of the conjunctivae was noted in all animals until observation on day 14. From day 14 onwards, staining of the conjunctivae was yellow. Staining of the sclera was evident in all animals on day 21 after application. The corneas of all treated animals were unaffected.

No corrosion was observed at any of the measuring intervals. Thus, the substance is considered non-irritant to rabbit eyes.