(2,2'-(3,3'-dioxidobiphenyl-4,4'-diyldiazo)bis(6-(4-(3-(diethylamino)propylamino)-6-(3-(diethylammonio)propylamino)-1,3,5-triazin-2-ylamino)-3-sulfonato-1-naphtholato))dicopper(II) acetate lactate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From November 26, 1990 to December 10, 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

February 24, 1987

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(2,2'-(3,3'-dioxidobiphenyl-4,4'-diyldiazo)bis(6-(4-(3-(diethylamino)propylamino)-6-(3-(diethylammonio)propylamino)-1,3,5-triazin-2-ylamino)-3-sulfonato-1-naphtholato))dicopper(II) acetate lactate

EC Number:

407-240-9

EC Name:

(2,2'-(3,3'-dioxidobiphenyl-4,4'-diyldiazo)bis(6-(4-(3-(diethylamino)propylamino)-6-(3-(diethylammonio)propylamino)-1,3,5-triazin-2-ylamino)-3-sulfonato-1-naphtholato))dicopper(II) acetate lactate

Cas Number:

159604-94-1

Molecular formula:

C66H88Cu2N20O10S2.C3H5O3.C2H3O2

IUPAC Name:

7,7'-bis[4-(3-diethylaminopropylamino)-6-(3-diethylammoniopropylamino)-1,3,5-triazin-2-ylamino]-{μ-4,4'-dihydroxy-1:2k2O4:O4'-3,3'-[3,3'-dihydroxy-1:2k2O3:O3'-biphenyl-bisazo-1:2(N3,N4-η:N3',N4'-η)]dinaphthalene-2-sulphonato(6-)}dicuprate(2-), mixed (1:1) acetic/lactic acid salts

Test material form:

solid

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Broekman Institute, Someren, The Netherlands
- Age at study initiation: approx. 19 weeks
- Weight at study initiation: 3093 - 3312 grams
- Housing: individually in cages with perforated floors and equipped with an automatic drinking system
- Diet: approx. 100 g per day
- Water: ad libitum
- Acclimation period: at least five days before start of treatment under test conditions

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C
- Humidity (%): 55%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hours artificial flourescent light / 12 hours dark per day

Test system

Vehicle:

unchanged (no vehicle)

Remarks:

The powdery test substance was instilled as delivered by the sponsor.

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 65 mg per animal

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

1, 24, 48 and 72 hours and 7 and 14 days

Number of animals or in vitro replicates:

3 female

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): solution of 2% fluorescein in water (adjusted to pH 7.0)
- Time after start of exposure: 24 hours

SCORING SYSTEM: Draize score
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

TOOL USED TO ASSESS SCORE: standard light/pocket flash light

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal: 325

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 2

Irritation parameter:

chemosis score

Basis:

animal: 315

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 3

Irritation parameter:

chemosis score

Basis:

animal: 305

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 14 days

Results 4

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: 325

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 5

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: 315

Time point:

24/48/72 h

Score:

2.3

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 6

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal: 305

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 14 days

Results 7

Irritation parameter:

iris score

Basis:

animal: 315

Time point:

24/48/72 h

Score:

0.33

Max. score:

2

Reversibility:

fully reversible within: 48 hours

Results 8

Irritation parameter:

iris score

Basis:

animal: 305/325

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible within: 48 hours

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal: 315

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 10

Irritation parameter:

cornea opacity score

Basis:

animal: 305/325

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Instillation of the test item into one of the eyes of each of three albino rabbits affected the cornea and the iris of animal 315 and affected the conjunctivae of all three animals.
The opacity of the cornea was reversible within 7 days and the injection of the iris within 48 hours in animal 315.
The irritation of the conjunctivae was reversible within 14 days in all three animals.
Treatment of the eyes with 2% fluoresce in revealed corneal epithelial damage at 24 and 72 hours after test article instillation in animal 315.
See results in "Any other information on results incl. tables"

Other effects:

Colouration: Blue discolouration and/or adherence of test article in the treated eye and blue discolouration of the fur by the test article was observed in all three animals.

Corrosion: There was no evidence of ocular corrosion.

Toxic symptoms/mortality: No toxic symptoms were observed in the animals during the test period and no mortality occurred.

Any other information on results incl. tables

Animal no.	Observation time after pplication	Corneal opacity	Iris lesion	Conjunctivae redness	Chemosis
305	24 hours	0	0	2	1
	48 hours	0	0	2	0
	72 hours	0	0	1	0
	Mean value	0	0	1,7	0,3
315	24 hours	1	1	2	1
	48 hours	1	0	3	1
	72 hours	1	0	2	1
	Mean value	1	0,3	2,3	1
325	24 hours	0	0	2	1
	48 hours	0	0	2	0
	72 hours	0	0	2	0
	Mean value	0	0	2	0,3

Applicant's summary and conclusion

Interpretation of results:

other: Category 2 (irritating to eyes) based on CLP criteria

Conclusions:

Eye Irrit. 2, H319

Executive summary:

The purpose of this study was to assess the possible irritation or corrosion potential when a single dose of the test item was placed in the conjunctiva) sac of a rabbit eye.

This study was carried out in accordance with OECD Guideline No. 405, "Acute Eye Irritation / Corrosion" and EEC Directive 84/449/EEC, Part Be 5, "Acute Toxicity - Eye Irritation' ,

The test item  was instilled in one of the eyes of each of the three animals, followed by six observations at approximately 1, 24, 48 and 72 hours and 7 and 14 after test article administration.

Under the conditions of this study, the test item resulted in adverse effects of the cornea and iris in one animal and in adverse effects of the conjunctivae in all three animals.

The opacity of the cornea was reversible within 7 days and the injection of the iris was reversible within 48 hours in the one animal .

The irritation of the conjunctivae was reversible within 14 days in all three animals.

No ocular corrosion was observed in any of the animals.

No signs of systemic intoxication were observed during the study period.