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Toxicological information

Eye irritation

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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:
guideline study with acceptable restrictions
Remarks:
according to OECD 405, GLP not specified

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1984
Report date:
1984

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
Directive 79/831/EEC
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
Disodium 4-amino-3-[[4-[(2-amino-4-hydroxyphenyl)azo]phenyl]azo]-5-hydroxy-6-(phenylazo)naphthalene-2,7-disulphonate
EC Number:
270-629-5
EC Name:
Disodium 4-amino-3-[[4-[(2-amino-4-hydroxyphenyl)azo]phenyl]azo]-5-hydroxy-6-(phenylazo)naphthalene-2,7-disulphonate
Cas Number:
68460-07-1
Molecular formula:
C28H22N8O8S2.2Na
IUPAC Name:
disodium 4-amino-3-({4-[(2-amino-4-hydroxyphenyl)diazenyl]phenyl}diazenyl)-5-hydroxy-6-(phenyldiazenyl)naphthalene-2,7-disulfonate
Constituent 2
Chemical structure
Reference substance name:
Disodium 4-amino-3-[[4-[(2,4-diaminophenyl)azo]phenyl]azo]-5-hydroxy-6-(phenylazo)naphthalene-2,7-disulphonate
EC Number:
272-559-0
EC Name:
Disodium 4-amino-3-[[4-[(2,4-diaminophenyl)azo]phenyl]azo]-5-hydroxy-6-(phenylazo)naphthalene-2,7-disulphonate
Cas Number:
68877-33-8
Molecular formula:
C28H23N9O7S2.2Na
IUPAC Name:
disodium 4-amino-3-({4-[(2,4-diaminophenyl)diazenyl]phenyl}diazenyl)-5-hydroxy-6-(phenyldiazenyl)naphthalene-2,7-disulfonate

Test animals / tissue source

Species:
rabbit
Strain:
New Zealand White
Remarks:
KFM
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Kleintierfarm Madoerin AG, 4414 Fuellingsdorf, Switzerland
- Age at study initiation: 14 to 15 weeks
- Weight at study initiation: 2.5 to 2.7 kg
- Housing: iindividually in stainless steel cages equipped with an automatic cleaning and drinking system
- Diet (e.g. ad libitum): pelleted standard rabbit maintenance diet Kliba #341, batch 92/83 (Klingentalmuehle AG)
- Water (e.g. ad libitum): tap water
- Acclimation period: four days under test conditions, with a pre-treatment health examination by a veterinarian

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 55 ± 10 %
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: The right eye remained untreated and was used for control purposes.
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g
Observation period (in vivo):
72 hours
Number of animals or in vitro replicates:
3, two females and one male
Details on study design:
a single dose of 0.1 g was administered to the left eye of each animal. The right eye remained untreated and was used as the reference control respectively. The eyes were not rinsed.


SCORING SYSTEM:
the eyes of each animal were examined 1, 24, 48 and 72 hours after administration. The irritation was assessed according to the OECD guideline 405. the corrosive properties of the test item and the color of the treated eyes were described and recorded.


TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein
eye examinations were meda with a sllit-lamp 30 SL (C. Zeiss AG) and a Varta Cliotrix diagnostic lamp

Body weights were recorded at pre-test, day 1 and at termination of the study on a Mettler PK 4800 balance.
All animals were killed at the end of the observation period by an intraveneous injection of T61 (Hoechst) into the ear vein.


Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
of all animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not relevant since no signs of irritation recorded at any observation time
Remarks on result:
no indication of irritation
Irritation parameter:
iris score
Basis:
mean
Remarks:
of all animals
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not relevant since no signs of irritation recorded at any observation time
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal: 125 (M)
Time point:
24/48/72 h
Score:
0.67
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
conjunctivae score
Basis:
animal: 126 (F)
Time point:
24/48/72 h
Score:
0.67
Max. score:
3
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
conjunctivae score
Basis:
animal: 127 (F)
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 72 hours
Remarks:
score 1 was still evident after 72 hours; but signs of irritation are regarded to reverse within a prolonged observation period of 21 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
of all animals
Time point:
24/48/72 h
Score:
0.89
Max. score:
4
Reversibility:
fully reversible within: 72 hours
Irritant / corrosive response data:
Ocular Reactions

Blue coloured staining of the conjunctival membrane was noted in all animals at all observations.
No corneal or iridial effects were noted during the study.
Severe discharge was observed in all animals 1 hour after administration.
Moderate conjunctival irritation was noted in all treated eyes 1 hour after treatment with minimal conjunctival irritation noted at the 24 and 48 Hour observations.
2 treated eyes appeared normal at the 72-Hour observation, in one animal score 1 was still evident.

Any other information on results incl. tables

Table 1: individual results

rabbit no

evaluation interval

cornea opacity

Iris

conjunctivae

score

chemosis

score

discharge

staining

125 (M)

1 hour

0

0

1

2

x

x

126 (F)

 

0

0

1

2

x

x

127 (F)

 

0

0

1

2

x

x

125 (M)

24 hours

0

0

1

2

-

x

126 (F)

 

0

0

1

2

-

x

127 (F)

 

0

0

1

2

-

x

125 (M)

48 hours

0

0

1

0

-

x

126 (F)

 

0

0

1

1

-

x

127 (F)

 

0

0

1

1

-

x

125 (M)

72 hours

0

0

0

0

-

x

126 (F)

 

0

0

0

0

-

x

127 (F)

 

0

0

1

0

-

x

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test item has not to be classified as eye irritant according to Regulation (EC) no 1272/2008.
Executive summary:

The primary eye irritation potential of the test item was investigated according to OECD test guideline No. 405 and Method B.5. The test item was applied by instillation of 0.1 g into the left eye of each of three young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48, 72 hours after test item instillation. 

The mean score was calculated (separately) for each animal across three scoring times (24, 48 and 72 hours after instillation) for corneal opacity, iritis, redness and chemosis of the conjunctivae. No signs of irritation were recorded for corneal opacity and iritis in all animals. The mean score for chemosis was 0.89. The mean score for conjunctival reddening was 0.67 in two animals, for the remaining animal 1.0. 

The instillation of the test item into the eyes caused a severe discharge at the first examination after 1 hour. Blue coloured staining of the conjunctival membrane was noted in all animals at all observations.

With the exception of on female all signs of irritation were reversible and were no longer evident 72 hours days after treatment. Score 1.0 conjuctival reddening observed 72 hours after treatment in one animal is regarded to reverse within a prolonged observation period of 21 days.

No clinical signs of toxicity were observed.

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

According to the findings in this study, the test item does not meet the criteria for classification according to Regulation (EC) no 1272/2008.