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

Eye irritation

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 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

Test material

Test material information

Constituent 1

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-{2-[4-({4,6-bis[(3-sulfonatopropyl)sulfanyl]-1,3,5-triazin-2-yl}amino)-5-methoxy-2-methylphenyl]diazen-1-yl}naphthalene-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

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Results 2

Irritation parameter:

other: redness

Basis:

animal #1

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

3

Results 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

Results 4

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

Results 5

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Results 6

Irritation parameter:

other: redness

Basis:

animal #2

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

3

Results 7

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

2

Results 8

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

Results 9

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

4

Results 10

Irritation parameter:

other: redness

Basis:

animal #3

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

3

Results 11

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: Mean of observation at 24, 48 and 72 hours

Score:

0

Max. score:

2

Results 12

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.