Benzoic acid, 2,3,4,5-tetrachloro-6-cyano-, methyl ester, reaction products with 4-[2-(4-aminophenyl)diazenyl]-3-methylbenzenamine and methanol sodium salt (1:1)

Endpoint summary

Description of key information

Two studies in rabbits were performed to evaluate irritating or corrosive properties of the test material to skin or eyes. Application of the test substance onto skin or into eyes did not provoke irritation.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

not specified

Remarks:

prior to GLP implementation

Species:

rabbit

Strain:

Himalayan

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight: 1.5 - 2 kg
- Housing: individually in metal cages
- Diet: ad libitum standard rabbit food, NAFAG, Gossau SG
- Water: ad libitum
- Acclimation period: for a minimum of 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1
- Humidity (%): 55 ± 5
- Photoperiod (hrs dark / hrs light): 10 hour light cycle day

Type of coverage:

occlusive

Preparation of test site:

other: clipped back and flank, left side also abraded, right side not abraded

Vehicle:

unchanged (no vehicle)

Controls:

not specified

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

Duration of treatment / exposure:

24 hours

Observation period:

up to 48 hour after removal of the dressings, 72 hours after initiation of treatment

Number of animals:

6, 3 males, 3 females

Details on study design:

Two days before treatment the entire back and the flank of the rabbits were shaved with an electric clipper and immediately before treatment the shaven skin on the left side was slightly scarified. For treatment gauze patches of 2.5 x 2.5 cm with test material were applied to the prepared abraded and non abraded skin, in quantities of 0,5 g.

The patches were covered with an impermeable material and were fastened to the body of the rabbit with adhesive tape. The dressings were removed after a 24 hour exposure. The skin irritation was scored immediately and 48 hours after removal of the dressings (24 and 72 hours after initiation of treatment). Skin irritation was scored in accordance with the irritation score described below. The severity of skin irritation was rated as follows: A mean value of less than 2 indicated mild, from 2 to 6 moderate and above 6 severe skin irritation.

SCORING SYSTEM:
Erythema and eschar formation:
No erythema; 0
Very slight erythema (barely perceptible); 1
Well defined erythema; 2
Moderate to severe erythema; 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth); 4
Maximum possible erythema score: 4

Edema formation:
No edema; 0
Very slight edema (barely perceptible); 1
Slight edema (edges of area well defined by definite raising); 2
Moderate edema (raised approximately 1 mm); 3
Severe edema (raised more than 1 mm and extending beyond area of exposure); 4
Maximum possible edema score: 4

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact skin

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Intact skin

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Abraded skin

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

4

Remarks on result:

other: Abraded skin

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24 and 72 hours

Score:

0

Max. score:

8

Remarks on result:

other: No description in this study report if the test substance is moistened to ensure good skin contact.

Interpretation of results:

GHS criteria not met

Conclusions:

The test material is not an irritant to the rabbit skin.

Executive summary:

A study similar to OECD Guideline 404 was performed. The primary irritation index, a measure of the acute irritation to the rabbit skin was 0. This indicates that the test material is not an irritant to the rabbit skin.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of lukewarm water.

GLP compliance:

not specified

Remarks:

prior to GLP implementation

Species:

rabbit

Strain:

Himalayan

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 1.5 - 2 kg
- Housing: individually in metal cages
- Diet: standard rabbit food - NAFAG, Gossau SG, ad libitum
- Water: water ad libitum
- Acclimation period: minimum of 4 days
Only rabbits with normal ophthalmic findings were used for these tests.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1
- Humidity (%): 55 ± 5
- Photoperiod (hrs dark / hrs light): 10 hour light cycle day

Vehicle:

unchanged (no vehicle)

Controls:

other: right untreated eye

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g

Duration of treatment / exposure:

In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of lukewarm water.
In the other 3 animals the eyes were not rinsed, therefore, they were exposed up to 7 days.

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

6: 3 male, 3 female

Details on study design:

The test material in an amount of 0.1 g was inserted into the conjunctival sac of the left eye of the 6 rabbits and the lids were gently closed for one second. The right eye was not treated and served as an untreated control. In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of lukewarm water. The eye irritation was appraised with a slit-lamp on day 1, 2, 3, 4 and 7 and was scored for each individual rabbit as outlined below. The primary eye irritation index represents the sum of the mean values for cornea, iris and conjunctivae. This procedure was carried out separately for unrinsed and rinsed eyes respectively.

SCORING SYSTEM:
CORNEA:
A Opacity and degree of density (most dense area scored):
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
A X B X 5
Maximum possible score = 80

IRIS:
A Values:
Normal, 0
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, 1
No reaction to light, hemorrhage, gross destruction (any or all of these), 2
A x 5
Maximum possible score = 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) X 2
Maximum possible score = 20

The total maximum score was thus 80 for the cornea, 10 for the iris, 20 for the conjunctivae and 110 for the entire eye. The severity of overall eye irritation was rated as:
0 - 10 minimal
11 - 25 slight
26 - 56 moderate
57 - 84 marked
above 84 extreme
The ratio R= unrinsed eyes/rinsed eyes, indicates the effect of rinsing the eyes 30 seconds after instillation of the compound.

<1 = rinsing increased the effect of the compound
1.1 - 1.5 = practically no effect
1.6 - 2 = little but assessable effect
2.1 - 4 = good effect
4.1 - 10 = very good effect
>10 = action of the compound practically abolished

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1 - 7 days

Score:

0

Max. score:

80

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 - 7 days

Score:

0

Max. score:

10

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 1 - 7 days

Score:

0

Max. score:

20

Remarks on result:

other: Unrinsed eyes

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1 - 7 days

Score:

0

Max. score:

80

Remarks on result:

other: Rinsed eyes

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 - 7 days

Score:

0

Max. score:

10

Remarks on result:

other: Rinsed eyes

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: 1 - 7 days

Score:

0

Max. score:

20

Remarks on result:

other: Rinsed eyes

Irritant / corrosive response data:

In the present experiments the irritation index was established as 0 for the cornea, 0 for the iris and 0 for the conjunctivae. The test material caused therefore no eye irritation to rabbits.

Interpretation of results:

GHS criteria not met

Conclusions:

The test material was found to cause no irritation.

Executive summary:

A study similar to OECD Guideline 405 was performed. Under the conditions of this experiment the test material was found to cause no irritation when applied to the rabbit eye mucosa.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Six rabbits (3/sex/dose) were each administered a single dermal dose of 0.5 g of the test substance. Occlusive application of the test material to intact rabbit skin for four hours did not cause any reaction on animals skin.

In an eye irritation test the test material, in an amount of 0.1 g, was inserted into the conjunctival sac of the left eye of the 6 rabbits ( 3 male/ 3 female) and the lids were gently closed for one second. The right eye was not treated and served as an untreated control. In 3 of the 6 rabbits approximately 30 seconds after treatment the treated eye was flushed with 10 mL of lukewarm water. The eye irritation was appraised with a slit-lamp on day 1, 2, 3, 4 and 7 and was scored for each individual rabbit. The primary eye irritation index represents the sum of the mean values for cornea, iris and conjunctivae. This procedure was carried out separately for unrinsed and rinsed eyes respectively. The irritation index was established as 0 for the cornea, 0 for the iris and 0 for the conjunctivae in unrinsed and rinsed eyes.

Application of the test substance onto skin did not cause corrosion or irritation. Appllication of the test material into the conjunctival sac of the eye did also not cause any treatment releated effect.

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for eye irritation under Regulation (EC) No. 1272/2008.