Reaction mass of Trisodium 4-({4-chloro-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)-2-{[1-ethyl-2-hydroxy-4-methyl-6-oxo-5-(sulfonatomethyl)-1,6-dihydropyridin-3-yl]diazenyl}benzenesulfonate and Trisodium 4-({4-chloro-6-[(3-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)-2-{[1-ethyl-2-hydroxy-4-methyl-6-oxo-5-(sulfonatomethyl)-1,6-dihydropyridin-3-yl]diazenyl}benzenesulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experiment start date - 16 July 1992; Experiment completion date - 28 July 1992; Study completion date - 30 September 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 4-({4-chloro-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)- 2-{[1-ethyl-2-hydroxy-4-methyl-6-oxo-5-(sulfonatomethyl)-1,6-dihydropyridin-3- yl]diazenyl}benzenesulfonate

Cas Number:

106359-90-4

IUPAC Name:

Trisodium 4-({4-chloro-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)- 2-{[1-ethyl-2-hydroxy-4-methyl-6-oxo-5-(sulfonatomethyl)-1,6-dihydropyridin-3- yl]diazenyl}benzenesulfonate

Test material form:

solid: particulate/powder

Details on test material:

None

Specific details on test material used for the study:

Code number: FAT 40000/G
Batch number: 67/92
Purity: 79.5%
Appearance: solid
Solubility: miscible
Storage: room temperature
Expiration date: 01 June 1993.

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Dr. Karl Thomae GmbH, Postfach 1755, D-7950 Biberach an der Riss
- Age at study initiation:15 weeks
- Weight at study initiation: male : 2.9 kg females: 2.4 - 2.8 kg
- Housing: Individually in stainless steel cages (size: 86 x 54 x 33 cm) equipped with feed hoppers and drinking water bottles.
- Diet (e.g. ad libitum): Pelleted standard Kliba 341, Batch 69/92 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, CH-4303 Kaiseraugst), ad libitum. Results of analysis for contaminants are included in this report.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum. Results of bacteriological, chemical and contaminant analyses are included in this report.
- Acclimation period: Five days under laboratory conditions after veterinary examination. Only animals without any visual signs of illness were used for the study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3°C
- Humidity (%): 40-70 %
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours artificial fluorescent light (approx. 100 Lux)/12 hours dark, music during the light period.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 g of the test article

Observation period (in vivo):

1, 24, 48 and 72 hours and 7 days after administration.

Number of animals or in vitro replicates:

1 male
2 females

Details on study design:

SCORING SYSTEM:

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 reaction 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, 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

TOOL USED TO ASSESS SCORE: hand-slit lamp

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

3 animals

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

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.88

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

3 animals

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within:

Remarks:

48 h

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

FAT 40'000/G showed a primary irritation score of: 1.67, when applied to the conjunctival sac of the rabbit eye. Only one animal was found to show opacity in the first hour of the test period. No corrosion was observed in all the other animals.

Other effects:

No acute systemic clinical symptoms were observed in the animals during the test and observation period, and no mortality occurred.

The lidhairs were yellowish dicolored in all animals at 1 hour after test article installation.

Female no. 61 lost weight (up to 8 %) during the aclimatization period. The body weight of the other animals was within the normal range of variability.

Any other information on results incl. tables

Observation table

OBESERVATION PERIOD	ANIMAL	SEX	CORNEA OPACITY	IRIS	CONJUNCTIVAE REDNESS	CONJUNCTIVAE CHEMOSIS
1 HR	59	M	1	0	1	1
	60	F	0	0	1	1
	61	F	0	0	2	2
24 HR	59	M	0	0	1	1
	60	F	0	0	1	1
	61	F	0	0	1	1
48 HR	59	M	0	0	1	0
	60	F	0	0	1	0
	61	F	0	0	1	0
72 HR	59	M	0	0	1	0
	60	F	0	0	1	0
	61	F	0	0	0	0
7 DAYS	59	M	0	0	0	0
	60	F	0	0	0	0
	61	F	0	0	0	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test item is considered to be "non-irritant" to the eyes.

Executive summary:

A GLP study was performed according to OECD test guideline 405 to determine the eye irritation potential of test substance.

The test substance was instilled in the eyes of 3 rabbits and observations for irritation were recorded at the time points of 1 hour, 24 hours, 48 hours 72 hours. 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. 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. The observation was made at 1h, 24h, 48h, 96h and 7 days timepoint.

The mean corneal opacity, iris, conjuctivae and chemosis score was found to be 0, 0, 0.88, 0.3 respectively. Based on the findings of this study, the test item is considered to be a "non-irritant" to the eyes.