Hydrogen [4-[[4-(diethylamino)-o-tolyl][4-[ethyl(3-sulphonatobenzyl)amino]phenyl]methylene]cyclohexa-2,5-dien-1-ylidene](ethyl)(3-sulphonatobenzyl)ammonium, sodium salt

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

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

Adequacy of study:

key study

Study period:

May 1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

The test was conducted by means of Read Across approach. Further information was attached at section 13

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1975

Report date:

1975

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965.)

Deviations:

yes

Remarks:

slight modification

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Acid Blue 015 - Similar Substance 01

IUPAC Name:

Acid Blue 015 - Similar Substance 01

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:

Animals:
Healthy New Zealand White rabbits (Porton strain) aged 10-13 weeks with average body weights of 2.71 kg (Males) and 2.66 kg (Females), bred on the premises and maintained under a semi-barrier system. Six rabbits (3 males and 3 females) were acclimatised in the test area for one week prior to the start of the trial.

Husbandry:
The rabbits were caged singly in an experimental room maintained at a temperature of 20 °C. (+/- 1.0 °C) and a relative humidity of 50-70 %. Animals
were exposed to artificial light for 10 hours daily from 08.00 - 18.00 hours. A commercial irradiated diet (Styles-Oxoid) was fed ad lib. Sterile filtered water was available at all times.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

100 mg of the test compound.

Duration of treatment / exposure:

One application.

Observation period (in vivo):

The rabbits were examined 1 and 6 hours, 1, 2, 3,6, and 10 days after application of the test compound.

Number of animals or in vitro replicates:

6 rabbits (3males and 3 females).

Details on study design:

The test compound were instilled into the conjunctival sac of the left eye 6 rabbits. The eyelids were then held closed for 1 second. The right eye served as a control. After 30 seconds, the Compound was, as far as possible, flushed out of the eyes of three of the rabbits with warm water for 1
minute.


SCORING SYSTEM

CORNEA:
(A) Opacity-degree of density (area most dense taken for reading)
*No opacity: 0
*Scattered or diffuse area, details of iris clearly visible: 1
*Easily discernible translucent areas, details of iris slightly obscured: 2
*Opalescent areas, no details of iris visible, size of pupil barely discernible: 3
*Opaque, iris invisible: 4

(B) Area of cornea involved
*One quarter (or less) but not zero: 1
*Greater than one quarter, but less than half: 2
*Greater than half, but less than three quarters: 3
*Greater than three quarters, up to whole area: 4

AxBx5 (Total maximun= 80)

IRIS:
(A) Value
*Normal: 0
*Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive): 1
*No reaction to light, hemorrhage, gross destruction (any or all of these): 2

Ax5 (Total maximum= 10)

CONJUNCTIVAE:
(A) Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
*Vessels normal: 0
*Vessels definitely injected above normal: 1
*More diffuse, deeper crimson red, individual vessels not easily discernible: 2
*Diffuse beefy red: 3

(B) Chemosis.
*No swelling: 0
*Any swelling above normal (includes nictitating membrane): 1
*Obvious swelling with partial eversion of lids: 2
*Swelling with lids about half closed: 3
*Swelling with lids about half closed to completely closed: 4

(C) Discharge
*No discharge: 0
*Any amount different from normal (does not include small amounts observed in inner canthus of normal animals: 1
*Discharge with moistening of the lids and hairs just adjacent to lids: 2
*Discharge with moistening of the lids and hairs, and considerable area around the eye: 3

(A+B+C)x2 (total maximum= 20)

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: mean 24-72h

Score:

0

Remarks on result:

other: Eyes washed

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: mean 24-72h

Score:

0

Remarks on result:

other: Eyes washed

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: mean 24-72h

Score:

0

Remarks on result:

other: Eyes washed

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: mean 24-72h

Score:

0

Remarks on result:

other: Eyes washed

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: mean 24-72h

Score:

0

Remarks on result:

other: Eyes washed

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: mean 24-72h

Score:

0

Remarks on result:

other: Eyes washed

Results 7

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: mean 24-72h

Score:

0.67

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Eyes washed

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

other: mean 24-72h

Score:

0.67

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Eyes washed

Results 9

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: mean 24-72h

Score:

0.33

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Eyes washed

Results 10

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: mean 24-72h

Score:

1

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Eyes washed

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: mean 24-72h

Score:

1

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Eyes washed

Results 12

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: mean 24-72h

Score:

1

Reversibility:

fully reversible within: 10 days

Remarks on result:

other: Eyes washed

Irritant / corrosive response data:

A mild to moderate conjunctival reaction was seen in all treated eyes one hour after application of the compound. This slowly subsided until all
eyes were normal by day 10.
A transient corneal reaction consisting of damage to the surface epithelium was seen in one eye at one hour only.
Unwashed eyes generally showed both a slightly more severe reaction than washed and staining of the conjunctivae by the compound for up to 48
hours.

Applicant's summary and conclusion

Interpretation of results:

other: CLP criteria not met

Conclusions:

Not eye irritant

Executive summary:

The purpose of this eye irritation study was to assess the possible irritation potential when single doses of test substance were placed in the conjunctival sac of rabbit left eyes (3 males and 3 female, 0.1 gr/animal).

This experiment was done according to the "Hazardous Substances Regulations" under the U.S.A. Federal Hazardous Substances Labelling Act Sect. 191.12 (February 1965.).

The test article (0.1 g per animal) was placed in the conjunctival sac of the left eye of each animal.

The right eye served as a control. After 30 seconds the compound was, as far as possible, flushed out of the eyes of three of the rabbits with warm water for 1 minute.

The rabbits were examined 1, 6, 24, 48 and 72 hours after application of the test compound and for any further period that was considered necessary.

Under the conditions of this experiment, the test substance was found to be not irritating when applied to the rabbit eye mucosa.