Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The reliability is rated 1 because the study followed the standard guideline of reference (OECD 405), which describes a procedure designed to evaluate this endpoint, the results were reviewed for reliability and assessed as valid, and the study was conducted under GLP condition.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

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:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3- sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

IUPAC Name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3- sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Constituent 2

Reference substance name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

EC Number:

700-610-3

Molecular formula:

UVCB substance - not applicable

IUPAC Name:

Bis{tris[4-(mono and dimethylamino)phenyl]methylium} 2-(bis{4-[ethyl(3-sulfonatobenzyl)amino]phenyl}methyliumyl)benzenesulfonate

Details on test material:

- Name of test material : Sepisol Fast Blue 2BR
- Physical state/ appearance: blue powder
- Lot/batch No.: 905536
- Storage condition of test material: ambient temperature and in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage GRIMAUD, (49450 Roussay, FRANCE)
- Weight at study initiation: 2.660 kg the day of the test material application
- Housing: Singly housed, in suspended stainless steel cage (60cm x 45 cm x 32 cm)
- Diet: granules Starlap Unic Robe (EVIALIS, 37018 Tours, France)
- Water : ad libitum, distributed by a polypropylene biberon with a stainless steel teat.
- Acclimation period: 5 days at least

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/-3
- Humidity (%): 50 +/- 20
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 hrs/1 2 hrs

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the untreated eye of the animal serves as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0,1 g

Duration of treatment / exposure:

single exposure followed by contention period of one hour

Observation period (in vivo):

15 days

Number of animals or in vitro replicates:

1 animal.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): No removal by washing.

SCORING SYSTEM: according to OECD 405 .

TOOL USED TO ASSESS SCORE: direct visualization with white light and examination with fluorescein.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean score: 24,48 and 72h after application

Score:

4

Max. score:

4

Reversibility:

not fully reversible within: D15

Remarks on result:

other: For ethical reasons, the study was stopped on D16 instead of D21.

Results 2

Irritation parameter:

other: Redness

Basis:

animal #1

Time point:

other: mean score: 24,48 and 72h after application

Score:

3

Max. score:

3

Reversibility:

not fully reversible within: D15

Remarks on result:

other: For ethical reasons, the study was stopped on D16 instead of D21.

Results 3

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: mean score: 24h, 48 and 72 h after application

Score:

1.7

Max. score:

2

Reversibility:

not fully reversible within: D15

Remarks on result:

other: For ethical reasons, the study was stopped on D16 instead of D21.

Results 4

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: mean score: 24h, 48h, 72h after application

Score:

2.7

Max. score:

3

Reversibility:

not fully reversible within: D15

Remarks on result:

other: For ethical reasons, the study was stopped on D16 instead of D21.

Irritant / corrosive response data:

The test item leads to a non-reversible irritation

Any other information on results incl. tables

SCORE

Chemosis:

Animal N°	Observation Period (after application)
	1h	24h	48h	72h	Mean*	Day 5 to Day 15	Day 16
2855	3	4	4	4	4.0	4	End of the study

 

Redness :

Animal N°	Observation Period (after application)
	1h	24h	48h	72h	Mean*	Day 5 to Day 9	Day 10 to Day 15	Day 16
2855	3	3	3	3	3.0	•	••	End of the study

 

Iris :

Animal N°	Observation Period (after application)
	1h	24h	48h	72h	Mean*	Day 5 to Day 9	Day 10 to Day 15	Day 16
2855	1	2	2	1	1.7	1	••	End of the study

 

Cornée :

Animal N°	Observation Period (after application)
	1h	24h	48h	72h	Mean*	Day 5 to Day 10	Day 11 to Day 15	Day 16
2855	0	3	3	2	2.7	2	••	End of the study

 

  *Mean score 24h, 48h, 72h after removal of the patch

  •Increase of the dark-blue coloration, red invaluable

•• Graduation impossible

Clinical observations:

Time Reading	Comments
	Animal #1
Day 1 (first hour)	Nothing to report
Day 2 up to day 8	Presence of a small amount of product in the conjunctival sac - Edematous and burned nictitating membrane - Opacity visible to the naked eye -Alopecia.
Day 9 up to Day 14	Presence of a small amount of product in the conjunctival sac - Edematous and burned nictitating membrane - Opacity visible to the naked eye -Alopecia - Glued eyes - Pus.
Day 15 up to Day 21	Eyes totally glued – End of the study by sacrifice of the animal for ethical reasons.

 

Applicant's summary and conclusion

Interpretation of results:

highly irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the experimental conditions, the test item is classified as irritant category 1 according to the CLP regulation (EC 1272/2008).

Executive summary:

The aim of this study was to assess qualitatively and quantitatively irritancy or corrosion and the delay of appearance of the effects after single application of 0.1 g of the pure test material named Sepisol Fast Blue 2BR as supplied on eye in one rabbit. The assay was

performed according to the OEDC guideline n° 405.

Ocular reactions (redness and chemosis of conjunctivae, iris and cornea lesions) were scored 1h, 24h, 48h and 72 h after application and until D15. The untreated eye served as control.

The results for the mean score were as follows (mean score 24h, 48h and 72h after application): chemosis: 4.0; redness: 3.0; Iris:1.7 and Cornea: 2.7.

In regards of the results observed and the irreversibility and the seriouness of the lesions, the study was ended at D15 for ethical reasons. Likewise she was not carried out on 2 other rabbits.

According to the criteria defined in the CLP regulation (EC 1272/2008), the test material is found to be classified as eye damage category 1.