Reaction mass of 5,5'-{(phenylmethanediyl)bis[benzene-4,1-diyl-diazene-2,1-diyl]}bis{1-[3-(dimethylamino)propyl]-4-methyl-6-oxo-3-(pyridinium-1-yl)-1,6-dihydropyridin-2-olate} hydrochloride and 5,5’-[3,4’-(phenylmethanediyl)diphenylene]bis(diazene-2,1-diyl)bis{1-[3-(dimethylamino)propyl]-4-methyl-6-oxo-3-(pyridinium-1-yl)-1,6-dihydropyridin-2-olate} hydrochloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2007-10-22 to 2008-01-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline-conform study under GLP without deviations, conducted with the analogue substance.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

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

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

analogue substance (refer to IUCLID chapter 13)

IUPAC Name:

analogue substance (refer to IUCLID chapter 13)

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands BV, Kreuzelweg 53, NL-5961 NM Horst / The Netherlands, Postbus 6174, NL-5960 AD Horst / The Netherlands
- Age at study initiation: 13 weeks (male), 13 - 14 weeks (females)
- Weight at study initiation: First Day of Acclimatization: 2187, 2523, 2418 g; Last Day of Observation: 3021, 3411, 3444 g
- Housing: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls.
- Diet (e.g. ad libitum): Pelleted standard rabbit maintenance diet ad libitum
- Water (e.g. ad libitum): Community tap water ad libitum.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2007-10-24 To: 2007-11-20

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

100 mg

Duration of treatment / exposure:

21 days

Observation period (in vivo):

21 days

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM: The eye reactions were assessed according to the numerical scoring system listed in the Commission Directive 2004/73/EC, April 29, 2004 (study report page 30). Scleral reddening and ocular discharge were also assessed.

TOOL USED TO ASSESS SCORE: hand-slit lamp

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

degree of corneal opacity

Basis:

mean

Remarks:

animals no. 1, 2, and 3

Time point:

other: overall at 24, 48, and 72 h

Score:

1

Max. score:

4

Reversibility:

other: staining not fully reversible within 21 days in one female,

Remarks on result:

other: One female with a very slight opacity of the cornea on day 21. Another test animal showed fully reversible reaction within 21 days, whereas the third test animal recovered fully within 7 days.

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

animals no. 1, 2, and 3

Time point:

other: overall at 24, 48, and 72 h

Score:

0.44

Max. score:

2

Reversibility:

fully reversible within: 2 days or 7 days, respectively

Results 3

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

mean

Remarks:

animals no. 1, 2, and 3

Time point:

other: overall at 24, 48 and 72 h

Score:

2

Max. score:

3

Reversibility:

fully reversible within: 10 days or 14 days, respectively

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animals no. 1, 2, and 3

Time point:

other: overall at 24, 48 and 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 5

Irritation parameter:

other: sclera

Basis:

mean

Remarks:

animals no. 1, 2, and 3

Time point:

other: overall at 24, 48, and 72 h

Score:

1.66

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity were 1.00 for each of the three animals and for iris were 0.00, 1.00 and 0.33, respectively. The individual mean scores for the conjunctivae were 2.00 for reddening and 1.00 for chemosis, for each of the three animals.
One hour after instillation, the opacity of the cornea and the reddening of the conjunctivae as well as the sclerae of the three animals were not assessable due to a marked yellow staining produced by the test item. When assessable at the 24-hour reading, a very slight opacity of the cornea affecting the whole area was observed in all animals. The opacity persisted up to day 10 and 21, respectively affecting more than one quarter or more than half of the cornea and was due to the staining caused by the test item. The iris of both females showed a delayed / reduced light reflex at the 24-hour observation up to the 72-hour reading in one female. Additionally, a moderate reddening of the conjunctivae was noted in all animals 24 hours after treatment and persisted as slight up to day 7 or 10, respectively. An obvious swelling with partial eversion of the lids was recorded in the three animals at the 1-hour observation and persisted as slight swelling up to the 72-hour evaluation. Furthermore, a slight to moderate reddening of the sclerae was present in the treated animals 24-72 hours after treatment.
Moderate ocular discharge was noted in all animals 1 hour after instillation and persisted as slight discharge up to the 72-hour observation.
No abnormal findings were observed in the treated eye of two animals 10 and 21 days after treatment, respectively. One female was still observed with a very slight opacity affecting more than half of the cornea on day 21, i.e. one of three animals showed persistent corneal effect.

Other effects:

MORTALITY/CLINICAL SIGNS: No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.

COLORATION:
A marked yellow staining produced by the test item was observed in each animal one hour after instillation and persisted as slight up to day 21.

CORROSION:
No corrosion of the cornea was observed at any of the reading times. BODY WEIGHTS: The body weights of all rabbits were considered to be within the normal range of variability.

TEST ITEM REMNANTS:
Yellow test item remnants were evident in the eye or conjunctival sac of each animal 1 hour after treatment and persisted up to the 24- or 72-hour reading.

Applicant's summary and conclusion

Interpretation of results:

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

Remarks:

Migrated information

Conclusions:

One of three test animals showed a persistent corneal effect (i.e. staining) which triggers classification for eye damage. Given the applicability of the proposed read across approach (see IUCLID chapter 13) the same classification also apply to the substance registered.

Executive summary:

The primary eye irritation potential of the analogue substance was being investigated following the testing protocol as given in OECD guideline 405.

The analogue substance 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 and 72 hours, as well as 7, 10, 14, 17 and 21 days after test item instillation.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after instillation) for each animal for corneal opacity, iris, redness and chemosis of the conjunctivae, separately. The individual mean scores for corneal opacity were 1.00 for all three animals and for iris were 0.00, 1.00 and 0.33, respectively. The individual mean scores for the conjunctivae were 2.00 for reddening and 1.00 for chemosis, for each of the three animals.

 

The instillation of the analogue substance into the eye resulted in mild to moderate, early-onset and transient ocular changes, such as very slight opacity of the cornea, delayed/reduced light reflex, reddening of the conjunctivae and sclerae, discharge and chemosis. These effects were reversible in two animals and were no longer evident at the latest 10 and 21 days after treatment, respectively. One female was still observed with a very slight opacity affecting more than half of the cornea on day 21, the end of the observation period. No corrosion was observed at any of the measuring intervals. A marked yellow staining of the treated eyes by the test item was observed which persisted as slight up to day 21, the end of the observation period. Yellow test item remnants were evident in the eye or conjunctival sac of each animal 1 hour after treatment and persisted up to the 24- or 72 hours. No clinical signs were observed.

One of three test animals showed a persistent corneal effect (i.e. staining) which triggers classification for eye damage. Given the applicability of the proposed read across approach (see IUCLID chapter 13) the same classification also apply to the substance registered.