4,4'-(3-cyano-4-methylpyridine-2,6-diyl)bis(azanediyl)dibenzenesulfonic acid

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Remarks:

enucleated eye nucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed on 21 April 2010. Report issued: 24 August 2010.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Rabbit Enucleated Eye Test (REET)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-({5-cyano-4-methyl-6-[(4-sulfophenyl)amino]pyridin-2-yl}amino)benzene-1-sulfonic acid

EC Number:

687-668-2

Cas Number:

815676-09-6

Molecular formula:

C19H16NO6S2

IUPAC Name:

4-({5-cyano-4-methyl-6-[(4-sulfophenyl)amino]pyridin-2-yl}amino)benzene-1-sulfonic acid

Constituent 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

water

impurity 1

Reference substance name:

Tetrahydrothiophene 1,1-dioxide

EC Number:

204-783-1

EC Name:

Tetrahydrothiophene 1,1-dioxide

Cas Number:

126-33-0

Molecular formula:

C4H8O2S

IUPAC Name:

Sulfolane

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

A volume of 0.1 ml of the test substance was applied, which was found to weigh approximately 85 mg (as measured following gently compacting the required volume into an adapted syringe).

Duration of treatment / exposure:

The test material was sprinkled as evenly as possible over the surface of the cornea. After 10 seconds the test material was washed off the cornea using a minimum of 20 ml of saline solution at approximatley 32°C. Untreated eye were similarly washed and used for control purposes.

Number of animals or in vitro replicates:

A total of five eye were used on the study. Three eyes treated with test substance and two eyes remained untreated for control purposes.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Remarks:

(cloudiness x area)

Value:

ca. 3

Negative controls validity:

valid

Results 2

Irritation parameter:

other: fluorescein uptake

Remarks:

(Intensity of fluorescein uptake x area)

Value:

ca. 3

Negative controls validity:

valid

Results 3

Irritation parameter:

other: condition of corneal epithelium

Negative controls validity:

valid

Remarks on result:

other: pitting

Remarks:

meets or exceeds cut-off value indicating a severe ocular irritant.

Results 4

Irritation parameter:

other: % corneal swelling of test eyes at 60 minutes

Value:

10.2

Negative controls validity:

valid

Results 5

Irritation parameter:

other: % corneal swelling of test eyes at 120 minutes

Value:

ca. 15.3

Negative controls validity:

valid

Results 6

Irritation parameter:

other: % corneal swelling of test eyes at 240 minutes

Value:

ca. 20.5

Negative controls validity:

valid

Results 7

Irritation parameter:

other: % corneal swelling of control eyes at 60 minutes

Value:

ca. 5.7

Results 8

Irritation parameter:

other: % corneal swelling of control eyes at 120 minutes

Value:

ca. 4.6

Results 9

Irritation parameter:

other: % corneal swelling of control eyes at 240 minutes

Value:

ca. 3.6

Other effects / acceptance of results:

Corneal Opacity
Some loss of transparency was noted in all test eyes. No corneal effects were noted in the control eyes during the study.

Corneal thickness and Condition
Corneal swelling of the test eyes was slightly greater than that observed in the control eyes over the same period. The condition of the corneal epithelium of the test eyes appeared normal 60 minutes after dosing but pitting was noted 120, 180 and 240 minutes after dosing. The condition of the corneal epithelium of the control eyes appeared normal during the study.

Fluorescein Uptake
Slight fluorescein staining was noted in the test eyes 240 minutes following test material application. No fluorescein uptake was noted in the control eyes 240 minutes following treatment.

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

Following assessment of the data for all endpoints, the test material was considered to have the potential to cause severe ocular irritancy in vivo. The in vivo eye irritation study is therefore not required.

Executive summary:

Introduction

The potential of the test substance to cause eye irritation/damage was assessed using the Rabbit Enucleated Eye Test (REET) due to the low pH of the test material, prior to possible in vivo testing.  This step-wise procedure is in accordance with OECD Test Guideline 405 and UK Home Office Regulations.  

Method

0.1 ml of the test material was applied onto the cornea of each of three enucleated eyes which had been maintained at a temperature of 32°C ± 1.5°C within a superfusion chamber. A further two enucleated eyes were treated, for control purposes, with saline solution (0.9% Sodium Chloride).

Results and Conclusion

Following assessment of the data for all endpoints the test material was considered to have the potential to cause severe ocular irritancy in vivo. The in vivo eye irritation study is therefore not required.