[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2012-06-25 to 2012-07-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

EC Number:

265-449-9

EC Name:

[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

Cas Number:

65113-55-5

Molecular formula:

C25H30N3.C18H14N3O3S

IUPAC Name:

[4-[p,p'-bis(dimethylamino)benzhydrylidene]cyclohexa-2,5-dien-1-ylidene]dimethylammonium m-[[p-anilinophenyl]azo]benzenesulphonate

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage de Gérome (Quartier Labaste - F40260 Linxe)
- Age at study initiation: 13 weeks old
- Weight at study initiation: 2.90 kg (beginning of the test) - 3.08 kg (end of the test)
- Housing: individual box installed in conventional aire conditioned animal husbandry
- Diet (e.g. ad libitum): SDS-C15, ad libitum
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C +/- 3 °C
- Humidity (%): 30 to 70%
- Air changes (per hr): 10 to 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12h light/12g dark

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g

Duration of treatment / exposure:

24 hr

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

1 animal

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): physiological rinse of the remaining test item
- Time after start of exposure: 1h

SCORING SYSTEM: Grading of Eye Reactions as displayed in the attached document titled "Solvent Black 46 - IO-OCDE-PH-12/0297 Scoring System".

TOOL USED TO ASSESS SCORE: fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal: A2132

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Results 2

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: A2132

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

fully reversible

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal: A2132

Time point:

24/48/72 h

Score:

2

Max. score:

2

Reversibility:

not reversible

Results 4

Irritation parameter:

iris score

Basis:

animal: A2132

Time point:

24/48/72 h

Remarks on result:

not determinable

Remarks:

the iris was not discernible at all reading times

Irritant / corrosive response data:

The test item leads to a non-reversible irritation

Other effects:

Blue coloration
White coloration of the upper membrane on day 7
Discharge with moistening of the eyelids and neighbouring hairs
Corneal neovascularization and corneal oedema
Iris not discernible at all reading times.

Any other information on results incl. tables

Total and Individual scores of ocular irritation

Observation time	CONJUNCTIVAE			IRIS	CORNEA
	Chemosis	Discharge	Redness	Iris	Degree of opacity	Extend of opacity
1 hour	2	2	1	*	0	0
24 hours	3	2	2	*	2	4
48 hours	3	2	2	*	2	4
72 hours	3	1	2	*	2	4
Day 7^•	3	1	2	*	2	4
Day 14^••	3	1	2	*	2	4
Day 21^••	3	2	0	*	2	4

 

*Iris not discernible

^•White coloration of the upper membrane

^••Corneal neovascularization and corneal oedema

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Under the experimental conditions, the test item is classified as irritant category 1 (irreversible effects on the eye) according to the CLP regulation (EC) No. 1272/2008.

Executive summary:

The test item was instilled as supplied, into the eye of one New Zealand rabbit at the dose of 0.1g. Remaining test item requiring a physiological rinse was noted one hour after the test item instillation. A blue coloration remained after rinsing. The experimental protocol was established on the basis of the official method as defined in the OECD guideline No. 405 dated April 24th, 2002 and the test method B.5 of the council regulation No. 440/2008.

 

The ocular reactions observed during the study have been slight to important and partially reversible:

- at the conjunctivae level: a slight redness noted 1 hour after the test item instillation, turning into a moderate redness 24 hours after the test item instillation and totally reversible on day 21, associated with a moderate chemosis noted 1 hour after the test item instillation, turning into an important chemosis 24 hours after the test item instillation and remaining on day 21 (last day of the test, same intensity).

-at the corneal level: a moderate corneal opacity, noted 24 hours after the test item instillation and remaining on day 21 (same intensity).

The iris was not discernible whatever the reading time.

A white coloration of the upper membre was noted on day 7.

A corneal neovascularization and a corneal oedema were noted on day 14 and remained on day 21.

 

In conclusion, taking into account the irreversibility of lesions observed, the results obtained, under these experimental conditions, enable to conclude that the item must be classified R41 "Risk of serious damage to eyes", according to the criteria for the classification, packaging and labelling of dangerous substances in complicance with E.E.C Directives 67/548, 2001/59 and 99/45. It must be characterised by the symbol "Xi" and the danger label "irritant".

 

In accordance with the Regulation (EC) No. 1272/2008, the test item must be classified in category 1 "irreversible effects on the eye". The signal word "Danger" and hazard statement H318 "Causes serious eye damage" are required.