Reaction products of diazotised 4,4-diaminodiphenylamine-2-sulfonic acid, subsequently coupled with 6-amino-4-hydroxynaphthalene-2-sulfonic acid, further diazotised and coupled with metaphenylendiamine, sodium salts

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Real Decree 363/1995 and Real Decree 1078/93

Principles of method if other than guideline:

Real Decree 363/1995 is the transposition of Directive 67/548/EEC in Spanish legislation

GLP compliance:

no

Remarks:

Centre Certificated ISO 9001 by National Accreditation Programme (certificate N. 98 - 1101)

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 6-[(2,4-diaminophenyl)azo]-3-[[4-[[4-[[7-[(2,4-diaminophenyl)azo]-1-hydroxy-3-sulphonato-2-naphthyl]azo]phenyl]amino]-3-sulphonatophenyl]azo]-4-hydroxynaphthalene-2-sulphonate

EC Number:

229-326-3

EC Name:

Trisodium 6-[(2,4-diaminophenyl)azo]-3-[[4-[[4-[[7-[(2,4-diaminophenyl)azo]-1-hydroxy-3-sulphonato-2-naphthyl]azo]phenyl]amino]-3-sulphonatophenyl]azo]-4-hydroxynaphthalene-2-sulphonate

Cas Number:

6473-13-8

Molecular formula:

C44H32N13O11S3Na3

IUPAC Name:

trisodium 6-[(2,4-diaminophenyl)azo]-3-[[4-[[4-[[7-[(2,4-diaminophenyl)azo]-1 hydroxy-3-sulphonato-2-naphthyl]azo]phenyl]amino]-3-sulphonatophenyl]azo]-4-hydroxynaphthalene-2-sulphonate

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: certificated provider
- Sex: males
- Housing: individual inoxidable steel cages, Tecniplast
- Diet: daily 150 g of special fodder for experimental rabbits, supplied by an authorized provider
- Water: automatic ad libitum, treated and filtered
- Marking: tatoo on the upper side of ear
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 2
- Humidity (%): 55 ± 25
- Air changes (per hr): 15 changes of pre-filtered air per hour
- Photoperiod (hrs dark / hrs light): 12/12

Other materials used:
Balance: COBOS D 6000-SX
Gloves: vinyl and latex gloves
Syringes: 1 mL of volume
Manual lamp
Wash bottle with saline solution
Colircusi fluorescein or equivalent authorized

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: in case of liquid substance were administrated 0.1 mL, in case of solid substance 0.1 g

Duration of treatment / exposure:

72 hours

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 males

Details on study design:

TOOL USED TO ASSESS SCORE: light of a manual lamp for examing the reactions and fluorescein for evaluating corneal lesions

SCORING SYSTEM:
Cornea
Opacity: degree of density (readings should be taken from most dense area)*
No ulceration or opacity: 0 Scattered or diffuse areas of opacity (other than slight dulling of normal lustre);
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
Maximum possible: 4
* The area of corneal opacity should be noted

Iris
Normal: 0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia;
or injection; iris reactive to light (a sluggish reaction is considered to be an effect: 1
Hemorrhage, gross destruction, or no reaction to light: 2
Maximum possible: 2

Conjunctivae
Redness (refers to palpebral and bulbar conjunctivae; excluding cornea and iris)
Normal: 0
Some blood vessels hyperaemic (injected): 1
Diffuse, crimson colour; individual vessels not easily discernible: 2
Diffuse beefy red: 3
Maximum possible: 3

Chemosis
Swelling (refers to lids and/or nictating membranes)
Normal: 0
Some swelling above normal: 1
Obvious swelling, with partial eversion of lids: 2
Swelling, with lids about half closed: 3
Swelling, with lids more than half closed: 4
Maximum possible: 4

DEFINITIONS:
1. Eye irritation is the production of changes in the eye following the application of a test substance to the anterior surface of the eye, which are fully reversible within 21 days of application.
2.Eye corrosion is the production of tissue damage in the eye, or serious physical decay of vision, following application of a test substance to the anterior surface of the eye, which is not fully reversible within 21 days of application.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0.11

Max. score:

0.33

Reversibility:

fully reversible

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0

Max. score:

0

Reversibility:

fully reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 1.22

Max. score:

1.33

Reversibility:

fully reversible

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0.66

Max. score:

1.33

Reversibility:

fully reversible

Any other information on results incl. tables

Results

	Rabbit 1						Rabbit 2								Rabbit 3
	1h	24h	48h	72h		7d	1h		24h		48h		72h	7d	1h	24h	48h	72h	7d
Corneal opacity	0	0	0	0		0	0		1		0		0	0	0	0	0	0	0
Iris	0	0	0	0		0	0		0		0		0	0	0	0	0	0	0
Conjunctival erythema	2	2	1	0		0	2		2		1		1	0	2	2	1	1	0
Conjuntival oedema	1	1	0	0		0	2		2		1		1	0	2	1	0	0	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Not an eye irritant

Executive summary:

Direct Black 22 was tested for eye irritation according to the Real Decree 363/1995 is the transposition of Directive 67/548/EEC in Spanish legislation.

0.1 g of TS were applied to three male New Zealand White rabbits, for 72 hours; observation period was of 7 days

Result

All lesions observed were completely reversible maximum in 72 hours.