Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
29 June 1995 to 02 July 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guidelines and the study was conducted under GLP conditions.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report date:
1995

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
Tetrasodium 3-[[4-[[4,6-bis-[(3-sulfonatoprop-1-yl)thio]-1,3,5-triazin-2-yl]amino]-2-methyl-5-methoxy-phenyl]azonaphthalene-1,5-disulfonate
EC Number:
421-950-6
EC Name:
Tetrasodium 3-[[4-[[4,6-bis-[(3-sulfonatoprop-1-yl)thio]-1,3,5-triazin-2-yl]amino]-2-methyl-5-methoxy-phenyl]azonaphthalene-1,5-disulfonate
Cas Number:
187674-70-0
Molecular formula:
C27H26N6Na4O13S6
IUPAC Name:
tetrasodium 3-[[4-[[4,6-bis-[(3-sulfonatoprop-1-yl)thio]-1,3,5-triazin-2-yl]amino]-2-methyl-5-methoxy-phenyl]azonaphthalene-1,5-disulfonate
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Substance type: yellow-brown to orange powder.
- Storage condition of test material: at room temperature.
- pH: 6.6 (10 % w/w in distilled water)

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 2.9 ± 0.2 kg
- Housing: Individually in polystyrene cages (35 x 55 x 32 cm or 48.2 x 58 x 36.5 cm).
- Diet: Pelleted diet ad libitum.
- Water: Filtered water ad libitum
- Acclimation period: At least 5 days.
- Animal selection: The day before treatment, the eyes of each animal were examined in order to use only animals without any signs of ocular irritation. Animals showing signs of ocular irritation, ocular defects or pre-existing corneal injury were not used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3 °C
- Humidity (%): 30 – 70 %
- Photoperiod: 12 hour light/dark cycle.

Test system

Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 mg per animal as supplied.
Duration of treatment / exposure:
A single exposure was performed.
Observation period (in vivo):
Animals were observed for 72 hours post application.
Number of animals or in vitro replicates:
3 male rabbits
Details on study design:
APPLICATION
- Method: The test material in its original form was placed into the conjunctival sac of the left eye of the 3 animals after gently pulling the lower lid away from the eyeball. The lower and upper lids were held together for about one second to avoid any loss of the test material. The right eye remained untreated as a control.

REMOVAL OF TEST SUBSTANCE
- Washing: Not performed.

SCORING SYSTEM: The eye was examined approximately 1 hour after administration and then at 24, 48 and 72 hours. The ocular reaction was scored according to the Draize scale, see Table 1. Any other lesions observed were noted.

TOOL USED TO ASSESS SCORE: Direct corneal examination was performed, if necessary with an Ultra Violet lamp. 1 or 2 drops of 0.5 % sodium fluorescein solution was used to determine the presence or absence of corneal opacification.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
other: redness
Basis:
animal #1
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
3
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal #1
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
other: redness
Basis:
animal #2
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
3
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal #2
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
4
Irritation parameter:
other: redness
Basis:
animal #3
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
3
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0
Max. score:
2
Irritation parameter:
cornea opacity score
Remarks:
(opacity)
Basis:
animal #3
Time point:
other: Mean of observation at 24, 48 and 72 hours
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 hours
Irritant / corrosive response data:
An orange colouration of the conjunctiva was noted 1 hour after treatment only, in all animals; redness of conjunctiva was marked by this colouration.
In one animal a slight (grade 1) corneal opacity was noted at the 24 hour reading only.
No other ocular lesions were observed during the study.

Any other information on results incl. tables

Table 2: Ocular Response

Rabbit No.

Region of Eye

Description of Ocular Reactions

Scores

Mean Irritation Score

1 hr

24 hrs

48 hrs

72 hrs

01

Conjunctivae

Chemosis

0

0

0

0

0.0

Redness

C

0

0

0

0.0

Discharge

0

0

0

0

0.0

Iris

 

0

0

0

0

0.0

Corneal Opacity

Intensity

0

0

0

0

0.0

Area

0

0

0

0

0.0

Other

 

*

*

*

*

 

Fluorescein

 

/

U

/

/

 

02

Conjunctivae

Chemosis

0

0

0

0

0.0

Redness

C

0

0

0

0.0

Discharge

0

0

0

0

0.0

Iris

 

0

0

0

0

0.0

Corneal Opacity

Intensity

0

0

0

0

0.0

Area

0

0

0

0

0.0

Other

 

*

*

*

*

 

Fluorescein

 

/

U

/

/

03

Conjunctivae

Chemosis

0

0

0

0

0.0

Redness

C

0

0

0

0.0

Discharge

0

0

0

0

0.0

Iris

 

0

0

0

0

0.0

Corneal Opacity

Intensity

0

1

0

0

0.3

Area

0

2

0

0

0.7

Other

 

*

*

*

*

 

Fluorescein

 

/

U

/

/

 

Mean score calculated from readings taken at 24, 48 and 72 hours.

* = None

C = Scoring masked by orange colouration

U = Fluorescein used

/ = Fluorescein not used

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the test, the test material was determined to be non-irritating, since a classifiable reaction was not observed in any of the treated rabbits.
Executive summary:

The acute eye irritation of the test material was determined in vivo in a study conducted under GLP conditions and in accordance with the standardised guidelines OECD 405 and EU method B.5.

Three male New Zealand White rabbits were exposed to 100 mg of the test material in a single ocular application. Ocular irritation was assessed 1 hour after application and then at 24, 48 and 72 hours, according to the Draize scale. Any other lesions were noted.

An orange colouration of the conjunctiva was noted 1 hour after treatment only, in all animals; redness of conjunctiva was obscured by this colouration. In one animal a slight (grade 1) corneal opacity was noted at the 24 hour reading only. No other ocular lesions were observed during the study.

Under the conditions of the test, the test material is considered to be a non-irritant since none of the animals displayed a reaction strong enough to trigger classification.