hexasodium 5,5'-{[2,7-disulfonato-9-(2-sulfonatophenyl)-9H-xanthene-9-ylium-3,6-diyl]bis(azanediyl)bis[(2,4,6-trimethyl-3,1-phenylene)aminosulfonyl]}diisophthalate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental 14-12-2016 to 23-12-2016. Final report issued 1-03-2017.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

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:

hexasodium 5,5'-{[2,7-disulfonato-9-(2-sulfonatophenyl)-9H-xanthene-9-ylium-3,6-diyl]bis(azanediyl)bis[(2,4,6-trimethyl-3,1-phenylene)aminosulfonyl]}diisophthalate

EC Number:

854-560-3

Cas Number:

1891094-25-9

Molecular formula:

C53H38N4Na6O22S5

IUPAC Name:

hexasodium 5,5'-{[2,7-disulfonato-9-(2-sulfonatophenyl)-9H-xanthene-9-ylium-3,6-diyl]bis(azanediyl)bis[(2,4,6-trimethyl-3,1-phenylene)aminosulfonyl]}diisophthalate

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

The New Zealand White rabbit is one of the standard strains used for acute irritation toxicity studies.
Animal health: Only animals in acceptable health condition were used for the test. Both eyes of each animal provisionally selected for testing were examined prior to starting the study. Animals showing eye irritation, ocular defects or pre-existing cornealinjury were not used.
Number of animals: 3 animals
Age of animals at treatment: 11 weeks old (young adult)
Sex: Male
Body weight range on the day of treatment: 2751 g – 2955 g before euthanasia: 2802 g – 3007 g
Date of receipt: 07 December 2016
Acclimatization time: at least 7 days
Animal identification: The individual identification was by engraved ear tag. The cages were marked with individual identity cards with information about study code, sex, dose, cage number and individual animal number.Animal health: Only healthy animals were used for the test. The veterinarian certified health status.
Number of animal room: 032
Light: 12 hours daily, from 6.00 a.m. to 6.00 p.m.
Temperature during the study: 19.54 – 22.53 °C
Relative humidity during the study: 29.56 – 55.82%
Housing/Enrichment: Rabbits were individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbit(s) in adjoining cages.
Ventilation: 15-20 air exchanges/hour The temperature and relative humidity values were measured continuously. The measured range was checked at least daily during the acclimatisation and experimental phases.
FOOD AND FEEDING Animals received UNI diet for rabbits produced by Cargill Takarmány Zrt., H-5300 Karcag,Madarasi út 0399, Hungary, ad libitum. Animals were provided with the following batches:
• 0003801145, expiry date: 01 March 2017
• 0003659483, expiry date: 03 January 2017

WATER SUPPLY AND QUALITY CONTROL OF WATER
The animals received municipal tap water, as for human consumption, ad libitum, from an automatic system. The quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8201 Veszprém, József A.u. 36., Hungary). The quality control results are retained in the archives of CiToxLAB HungaryLtd.

Test system

Vehicle:

water

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A single amount of 0.125 g of the solid (using the correction factor: 1.25) test item was powdered and administered as a single dose. The test substance was placed in the conjunctival sac of the left eye of each animal after
gently pulling the lower lid away from the eyeball. The lids were then gently held together for at least one second in order to prevent loss of the material. The untreated contralateral eye served as the control.

Duration of treatment / exposure:

The treated eye of test animals was rinsed with physiological saline solution, as the irritation scores more than one at one hour after application.

Observation period (in vivo):

Clinical Observations and Evaluation of Ocular Irritation
The eyes were examined at 1, 24, 48, 72 hours after treatment. The duration of the
observation period was sufficient to identify 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, the animal was sacrificed by intramuscular injections of Ketamin
10% (Ketamidor) and Xylazin 2% (Primazin 2%) followed by i.v. pentobarbital sodium (see
details in 3.2.). Death was verified by checking pupil and corneal reflex and the absence of
respiration.
All rabbits were examined for distress at least twice daily, with observations at least 6 hours
apart. Clinical observations or signs of ill-health were recorded.

Number of animals or in vitro replicates:

3

Details on study design:

The treated eye of test animals was rinsed with physiological saline solution, as the irritation scores more than one at one hour after application.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 3

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 4

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Results 5

Irritation parameter:

other: redness

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0 - < 0.33

Max. score:

0.33

Results 6

Irritation parameter:

other: discharge

Basis:

mean

Time point:

24/48/72 h

Score:

ca. 0 - < 0.66

Max. score:

0.66

Other effects:

As no clinical signs were observed, the experiment was terminated after 72 hours observation. During the experiment, the control eye of each animal was symptom-free.
The general state and behaviour of animals were normal throughout the study period.

Any other information on results incl. tables

Animal 1 (No: 6290) clinical observation

At 1 and 24 hours after the application, conjunctival redness (score 1) and discharge (score 2) were noted in the rabbit. The test item coloured the hairs to purple around the eye in the animal at 1 hour after the application. At 48 and 72 hours after the application, no clinical signs, and no conjunctival or corneal effects were observed.

Animal 2 (No: 6121) clinical observation

At 1 hour after the application, conjunctival redness (score 1) and discharge (score 2) were noted in the rabbit. The test item coloured the hairs to purple around the eye in the animal at 1 hour after the application.

At 24 hours after the application, conjunctival redness (score 1) and discharge (score 1) were noted in the rabbit.

At 48 and 72 hours after the application, no clinical signs, and no conjunctival or corneal effects were observed.

Animal 3 (No: 6122) clinical observation

At 1 hour after the application, conjunctival redness (score 1) and discharge (score 2) were noted in the rabbit. The test item coloured the hairs to purple around the eye in the animal at 1 hour after the application.

At 24,48 and 72 hours after the application, no clinical signs, and no conjunctival or corneal effects were observed.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance applied to rabbit eye mucosa, caused conjunctival effects at one and 24 hours after the treatment which were fully reversible within 48 hours.
There were no corneal effects observed in the study.
According to Regulation (EC) No 1272/2008, Dye-2016 does not require classification as an eye irritant.
According to the UN Globally Harmonised System of Classification and Labelling of Chemicals, test substance does not require classification as an eye irritant.

Executive summary:

Introduction

The objective of the study was to determine the acute eye irritation effect of the test item in the New Zealand White Rabbit. The test item was applied as a single dose to the left eye of treated animals. The degree of irritation was scored at specified time intervals.

Results

MORTALITY-There was no mortality observed during the study.

BODY WEIGHTS-The body weight of the animals was considered to be within the normal range of variability.

CLINICAL OBSERVATION

. General daily examination-There were no clinical signs observed that could be related to treatment.

No Initial Pain Reaction/Pain reaction (IPR/PR) was observed

Conclusion

The test substance, applied to rabbit eye mucosa, caused conjunctival effects at one and 24 hours after the treatment which were fully reversible within 48 hours.There were no corneal effects observed in the study.

According to Regulation (EC) No 1272/2008, Dye-2016 does not require classification as an eye irritant.

According to the UN Globally Harmonised System of Classification and Labelling of Chemicals, test substance does not require classification as an eye irritant.