2-Naphthalenesulfonic acid, 6-amino-5-[2-[4-[[4-[bis(2-hydroxyethyl)amino]-6-[(2-sulfoethyl)amino]-1,3,5-triazin-2-yl]amino]-2-sulfophenyl]diazenyl]-4-hydroxy-, sodium salt (1:3)

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 2 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, performed to 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:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

419-720-5

EC Name:

-

Cas Number:

182061-89-8

Molecular formula:

C25H26N9O12S3 . 3 Na

IUPAC Name:

trisodium 6-amino-5-{2-[4-({4-[bis(2-hydroxyethyl)amino]-6-[(2-sulfonatoethyl)amino]-1,3,5-triazin-2-yl}amino)-2-sulfonatophenyl]diazen-1-yl}-4-hydroxynaphthalene-2-sulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): M-377
- Physical state: solid
- Appearance: garnet powder
- Storage condition of test material: room temperature and protected from light
- pH of substance: 8.3 (measured at a concentration of 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
- Sex: males
- Weight at study initiation: 2.6 ± 0.2 kg (mean)
- Housing: individually in polystyrene cages
- Diet: pelleted diet, ad libitum
- Water: filtered drinking water (0.22 µm), ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 ± 3 °C
- Humidity (%): 30 - 70 %
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light

IN-LIFE DATES: From 29 June 1995 to 2 July 1995

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

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

Duration of treatment / exposure:

A single application was performed

Observation period (in vivo):

72 hours following dose administration

Number of animals or in vitro replicates:

3 males

Details on study design:

ADMINISTRATION
- Dose administration: Approximately 100 mg of the test material was applied into the conjunctival sac of the left eye of each rabbit by gently pulling the lower lid away from the eyeball. The lower and upper eyelids were then gently held together for about 1 second to avoid any loss of test material. The right eye was untreated (control eye).

REMOVAL OF TEST SUBSTANCE
- The eyes were not rinsed after administration of the test material.

SCORING SYSTEM
- The reactions observed were scored in accordance with the criteria of Draize (see field "Any other information on materials and methods incl. tables" for further information), at 1, 24, 48 and 72 hours after administration of the test material.

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:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: no effects observed

Results 2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.4

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Remarks on result:

other: no iridial effects noted throughout study

Results 4

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: no corneal effects noted throughout study

Irritant / corrosive response data:

Very slight or slight conjunctival redness (grade 1 or 2) was observed at the 24 hour reading in all animals. Conjunctival redness was masked by a garnet colouration of the conjunctiva one hour after treatment only; in addition, a moderate clear discharge (grade 2) was noted in one animal only. No effects on the iris or cornea were noted. No ocular reactions were observed after the 24 hour reading.

Any other information on results incl. tables

Table 2: Results

Rabbit number	Region of eye	Ocular reaction	Score				Mean irritation score**
			1 hour	24 hours	48 hours	72 hours
1	Conjunctivae	Chemosis	0	0	0	0	0.0
		Redness	C	1	0	0	0.3
		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	/	/
2	Conjunctivae	Chemosis	0	0	0	0	0.0
		Redness	C	2	0	0	0.7
		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	/	/
3	Conjunctivae	Chemosis	0	0	0	0	0.0
		Redness	C	1	0	0	0.3
		Discharge	2	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	/	/

** = Mean of readings taken at 24, 48 and 72 hours only

C = scoring obscured by garnet colouration

* = None

/ = fluorescein not used

U = fluorescein used

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the study, the test material only elicited slight reactions in any of the animals during the course of the study that meant the test material does not require classification as an eye irritant. The study is considered to be reliable, relevant and adequate for risk assessment and classification and labelling purposes.

Executive summary:

The eye irritation potential of the test material was determined in accordance with standardised guidelines OECD 405 and EU Method B.5. Approximately 100 mg of test material was applied into one eye of each of three rabbits; the animals were assessed for up to 72 hours to determine the grade of ocular reaction. Very slight or slight conjunctival redness (grade 1 or 2) was observed at the 24 hour reading in all animals. Conjunctival redness was masked by a garnet colouration of the conjunctiva one hour after treatment only; in addition, a moderate clear discharge (grade 2) was noted in one animal only. No effects on the iris or cornea were noted. No ocular reactions were observed after the 24 hour reading. Based on these observation it was determined that the test material in not classified for eye irritation.