Sodium 1-amino-9,10-dihydro-4-[5-[(2-hydroxyethyl)sulphamoyl]-3,4-xylidino]-9,10-dioxoanthracene-2-sulphonate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1981

Report date:

1981

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

FAT 20033/L TE

IUPAC Name:

FAT 20033/L TE

Constituent 2

Reference substance name:

Sodium 1-amino-9,10-dihydro-4-[5-[(2-hydroxyethyl)sulphamoyl]-3,4-xylidino]-9,10-dioxoanthracene-2-sulphonate

EC Number:

247-269-2

EC Name:

Sodium 1-amino-9,10-dihydro-4-[5-[(2-hydroxyethyl)sulphamoyl]-3,4-xylidino]-9,10-dioxoanthracene-2-sulphonate

Cas Number:

25797-81-3

Molecular formula:

C24H23N3O8S2.Na

IUPAC Name:

sodium 1-amino-4-({3-[(2-hydroxyethyl)sulfamoyl]-4,5-dimethylphenyl}amino)-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

powder / dark blue

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: EN 01451.12

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2 to 3 kg
- Housing: The animals were housed individually in metal cages
- Diet: Standard rabbit food - NAFAG, No. 814, Gossau SG (ad libitum)
- Water: ad libitum
- Acclimation period: 4 d

ENVIRONMENTAL CONDITIONS
- Temperature: 22±2 °C
- Humidity: 55±10 %
- Photoperiod (dark / light): 12 h/12 h

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the second eye on the treated animals served as control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 100 mg
- Concentration (if solution): Undiluted

Duration of treatment / exposure:

Unwashed eye: Duration of treatment same as observation period.
Washed eye: 30 seconds

Observation period (in vivo):

7 d

Number of animals or in vitro replicates:

6 (3 males and 3 females)

Details on study design:

PROCEDURE:
- Only rabbits with normal ophthalmic findings were used for the test.
- The test substance in an amount of 100 mg was inserted into the conjunctival sac of the left eye of the rabbits and the lids were gently closed for a few seconds. The right eye was not treated and served as an untreated control.
- In 3 of the 6 rabbits approx. 30 seconds after treatment the treated eye was flushed with 10 mL of physiological saline.
- The eye irritation was evaluated with a slit-lamp on 1, 2, 3, 4 and 7 d and was scored for each individual rabbit.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with 10 ml of physiological saline
- Time after start of exposure: 30 sec

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

B: Area of cornea involved
1 - One quarter (or less) but not zero
2 - Greater than one quarter, but less than half
3 - Greater than half, but less than three quarters
4 - Greater than three quarters, up to whole area
A x B x 5 Maximum possible score = 80

IRIS
A: Values
0 - Normal
1 - Folds above normal, congestion, swelling, circumcornea injection (any or all of these or combination of any thereof) iris still reacting to light (sluggish reaction is positive)
2 - No reaction to light, hemorrhage, gross destruction (any or all of these)
A x 5 Maximum possible score = 10

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

B: Chemosis
0 - No swelling
1 - Any swelling above normal (includes nictitating membrane)
2 - Obvious swelling with partial eversion of lids
3 - Swelling with lids about half closed
4 - Swelling with lids about half closed to completely closed
C Discharge
0 - No discharge 0
1 - Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2 - Discharge with moistening of the lids and hairs just adjacent to lids
3 - Discharge with moistening of the lids and hairs, and considerable area around the eye
(A + B + C) x 2 Maximum possible score = 20


TOOL USED TO ASSESS SCORE: hand-slit lamp

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.33

Max. score:

3

Reversibility:

not fully reversible within: 7 d

Results 2

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

3

Reversibility:

not fully reversible within: 7 d

Results 3

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

not fully reversible within: 7 d

Results 4

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

other: Test substance was washed after 30 seconds of application

Results 5

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #5

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 d

Remarks on result:

other: Test substance was washed after 30 seconds of application (however, observation period was up to 7 d only)

Results 6

Irritation parameter:

conjunctivae score

Remarks:

(redness)

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

3

Remarks on result:

other: Test substance was washed after 30 seconds of application

Results 7

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 7 d

Results 8

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.66

Max. score:

4

Reversibility:

not fully reversible within: 7 d

Results 9

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.66

Max. score:

4

Reversibility:

not fully reversible within: 7 d

Results 10

Irritation parameter:

chemosis score

Basis:

animal #4

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Test substance was washed after 30 seconds of application.

Results 11

Irritation parameter:

chemosis score

Basis:

animal #5

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

not fully reversible within: 7d

Remarks on result:

other: Test substance was washed after 30 seconds of application (however, observation period was up to 7 d only)

Results 12

Irritation parameter:

chemosis score

Basis:

animal #6

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

other: Test substance was washed after 30 seconds of application.

Results 13

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 14

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Irritant / corrosive response data:

The test substance was found to cause a minimal irritation (i.e., primary irritation index 5.7) when applied to the rabbit eye mucosa. When the application site was rinsed, the irritation was found to be minimal (i.e., primary irritation index 0.7).

Other effects:

none

Any other information on results incl. tables

Calculation of the primary eye irritation index:

	MEAN REACTION SCORE
Time after exposure (d)	Unrinsed eyes (A)			Rinsed eyes (B)
	cornea	iris	conjunctiva	cornea	iris	conjunctiva
1	0	0	8	0	0	1.3
2	0	0	6.7	0	0	1.3
3	0	0	5.3	0	0	0.7
4	0	0	5.3	0	0	0
7	0	0	3.3	0	0	0
subtotal	0	0	28.6	0	0	3.3
total	28.6			3.3

Primary irritation index in unrinsed eyes:

A = 28.6 : 5 = 5.7

Primary irritation index in rinsed eyes:

B = 3.3 : 5 = 0.7

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The test substance produced only slight conjunctival reaction.

Executive summary:

An in vivo study was conducted to evaluate the acute eye irritation potential of the test substance (of ca. 82.2% purity) according to a procedure similar to OECD Guideline 405 with few deviations. 100 mg of the test substance was placed into the conjunctival sac of the left eye of rabbits (4 male and 2 females). The right eye remained untreated and was used as untreated control. In three rabbits washing was done with 10 mL of physiological saline after 30 seconds of test substance instillation. Assessment of ocular damage/irritation was made with a slit-lamp approximately on 1, 2, 3, 4 and 7 d following treatment. No corneal or iridial effects were noted during the study. Minimal irritation was observed with primary irritation index of 5.7 and 0.7 for rinsed and unrinsed eyes respectively. Under the study conditions, the test substance produced only slight conjunctival reaction.