Pentasodium 2,2'-((2,4-diamino-5-(4-methoxy-2-sulfophenylazo)-1,3-phenylene)bis(azo))bis(5-(2-(sulfonatooxyethyl)sulfonyl)benzenesulfonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 04, 2006 to May 11, 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

475-300-1

EC Name:

-

Cas Number:

910030-59-0

Molecular formula:

Hill formula: C29H25N8Na5O22S7 CAS formula: C29H30N8O22S7.5Na

IUPAC Name:

Pentasodium 2,2'-((2,4-diamino-5-(4-methoxy-2-sulfophenylazo )-1,3-phenylene)bis(azo))bis(5-(2-(sulfonatooxyethyl)sulfony l)benzenesulfonate

Test material form:

solid: particulate/powder

Details on test material:

see below

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Ferenc Sandor breeder, 2173 Kartal, Császár út 135, Hungary
- Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined 24 h prior to the start of study. Animals showing eye irritation, ocular defects or pre-existing corneal injury were not used.
- Age of animals at arrival: 11 weeks old, adult albino rabbit
- Body weight range at treatment: 2614-3017 g (i.e., at the beginning of the study); 2739-3108 g (at the end of the study)
- Housing: Animals were housed individually in metal cages.
- Diet: PURINA Base – Lap gr. diet for rabbit, ad libitum
- Water: Tap water from self-supplying watering system, ad libitum
- Acclimation period: Male: 9 d
-Animal identification: The individual identification was performed by numbered ear tag. The cages were marked with individual identity cards with information about study code, sex, dose group, cage number and individual animal number

ENVIRONMENTAL CONDITIONS
- Temperature: 20±3°C
- Humidity: 30-70%
- Photoperiod: 12 h light/dark cycle

IN-LIFE DATES: From: To: May 04, 2006 to May 11, 2006

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg (i.e., 0.1 g)

Duration of treatment / exposure:

24 h

Observation period (in vivo):

1, 24, 48, 72 h and one week after the application of test substance

Number of animals or in vitro replicates:

3 males

Details on study design:

TEST PROCEDURE

Dosage
0.1 g of the test substance was used for the study in pure state, in a single dose. The absorption of the test substance was not determined.

Application of the Test Substance
Three male animals in acceptable health condition were selected for the test. Care was taken to select only those animals that had a normal eye condition, any with ocular lesions were rejected.

In the first step an initial test was performed using one animal. The test substance was instilled into the conjunctival sac of the left eye. The eyelids were held closed gently for several seconds to prevent the loss of the test substance. The contralateral eye served as control. Immediately after the administration of the test substance, an assessment of the initial pain reaction was made. After consideration of the ocular responses produced in the first animal, two additional animals were treated. Before the administration the treated eyes of animals were not anaesthetised, because the score of initial pain reaction was 0 in the first animal.

Duration of Exposure
The eyes of the test animals were not washed out after the application of test substance.

OBSERVATIONS AND SCORING

Clinical Observations

The eyes were examined at 1, 24, 48, 72 h then one week after the treatment. The duration of the observation period was sufficient for the statement of reversibility or irreversibility of changes. Any clinical signs of toxicity or signs of ill- health during the study were recorded. At the end of the observation period all animals were sacrificed by intramuscular injections of Ketamin and Xylazin followed by i. v. Euthanyl anaesthesia.

Scoring and Assessment of Local Reaction

The eye irritation scores were evaluated according to the scoring system by Draize (1959) and OECD 405 (24 April 2002).

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal: 1, 2, and 3

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

probability of mild irritation

Results 2

Irritation parameter:

chemosis score

Basis:

animal: 1, 2, and 3

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal: 1 and 3

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 d

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

cornea opacity score

Remarks:

(opacity)

Basis:

animal #2

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

iris score

Basis:

animal: 1, 2, and 3

Remarks:

(mean score)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

1h after treatment some hyperaemic blood vessels with obvious conjunctival swelling with partial eversion of the lids were found in every animal. In these animals the discharge moistened lids and hairs on considerable area around the treated eye. In all animals diffuse areas of opacity were observed on the cornea but the details of iris were clearly visible. Area of cornea involved was greater than three quarters, up to the whole area.

There were no effects observed in the iris alterations during the study period.

24 h after treatment in all animals some hyperaemic blood vessels were observed. Chemosis and increased discharge excretion were not found by this time. In two animals the degree and extension of cornea opacity did not change when compared to the previous observation. In another animal cornea opacity was not found by this time.

48 and 72 h after treatment in the animals the slight redness and degree and extension of cornea opacity did not change when compared to the previous observation.

One week after the treatment every animal was symptom-free.

One week after the treatment the study was terminated, since no primary irritation symptoms remained.

Other effects:

General state and the behaviour of animals were normal throughout the study period. There were no notable body weight changes during the contact and observation period.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Under the test conditions, the test substance was considered to be slightly irritating to eyes. Due to the 24/48/72 h mean cornea opacity score of >=1 in 2/3 animals, a classification according to CLP criteria applies. The substance is considered to cause irritation in eyes.

Executive summary:

A study was conducted to assess the eye irritancy potential of the test substance in male New Zealand White rabbits according to OECD Guideline 405, EPA OPPTS 870.2400 and EU Method B.5, in compliance with GLP.

A single dose of 100 mg test substance was applied to the conjunctival sac of the left eye of each animal. The treated eyes were not washed out after administration. The untreated eyes served in each case as a control.

Slight redness, moderate chemosis and severely increased discharge along with the diffuse corneal opacity were observed in the eyes of the treated animals at 1h after application. Further, slight redness and diffuse areas of opacity persisted until 72 h post-application. However, these irritation response were completely reversible at one week after the treatment. Furthermore, no effects were observed in the iris during the study period.

Under the test conditions, the test substance was considered to be slightly irritating to eyes.