6-(2-chloro-6-cyano-4-nitrophenylazo)-4-methoxy-3-[N-(methoxycarbonylmethyl)-N-(1-methoxycarbonylethyl)amino]acetanilide

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

6 January 1999 to 14 January 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP methodology followed and OECD Guideline 404 (Acute Dermal Irritation / Corrosion) used to performed the experiment.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Principles of method if other than guideline:

None

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

430-500-8

EC Name:

-

Cas Number:

204277-61-2

Molecular formula:

Hill formula: C23 H23 Cl N6 O8 CAS formula: C23 H23 Cl N6 O8

IUPAC Name:

methyl 2-({4-[2-(2-chloro-6-cyano-4-nitrophenyl)diazen-1-yl]-5-acetamido-2-methoxyphenyl}(2-methoxy-2-oxoethyl)amino)propanoate

Test material form:

other: solid

Details on test material:

None

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:New Zaeland White rabbit-Chbb:NZW(SPF); Elevage scientifique des Dombes, F-01400 Chatillon sur Chalaronne
- Age at study initiation: 15 weeks
- Identification: By unique cage number and corresponding ear number
- Accomodation: Individually in stainless steel cages with an automatic cleaning system equipped with feed hoppers, drinking water bowls and wood for gnawing.
- Diet: Pelleted standard Kliba 3410 rabbit maintenance diet ad libitum (batch n°79/97)
- Water: Community tap water from Itingrn, ad libitum, in water bowls.
- Acclimation period: 5 days under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C +/- 3°C
- Humidity (%): 40-70%
- Air changes (per hr): 10-15 ait changes per hour
- Photoperiod (hrs dark / hrs light): the room was illuminated by fluorescent light on a 12 hour light/dark cycle. Recorde music was played for approximately 8 hours during the light period

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

other: Bidistilled water

Controls:

no

Amount / concentration applied:

Approximately 0.5 gr (per animal, left side only)

Duration of treatment / exposure:

4 hours

Observation period:

3 days

Number of animals:

1 male and 2 females

Details on study design:

TEST SITE
On the day of treatment, the test articel was applied to approximately 6 cm2 of the intact skin of the clipped area. It was covered with a 2.5 cmX2.5 cm patch of surgical gauze and thegauze was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.

REMOVAL OF TEST SUBSTANCE
After the treatment (4 hours), the skin was flushed with lukewarm tap water to clean the application site.


SCORING SYSTEM:
ERYTHEMA and ESCHAR FORMATION:
- No erythema: 0
- Very slight erythema: 1
- Well-defined erythema: 2
- Moderate to severe erythema: 3
- Severe erythema (beet redness) or eschar formation (injuries in depth preventing erythema reading): 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 (edges raised approximatela 1 mm): 3
- Severe (raised more than 1 mm and extending beyond the area exposure)

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Remarks:

1 male and 2 females

Time point:

other: 1-72 hours

Score:

0

Results 2

Irritation parameter:

edema score

Basis:

mean

Remarks:

1 male and 2 females

Time point:

other: 1-72 hours

Score:

0

Results 3

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Remarks:

1 male and 2 females

Time point:

other: 1-72 hours

Score:

0

Other effects:

- Viability/Mortality and clinical signs:
No clinical signs of systemeic toxicity were observed in the animals during the test and observation period, and no mortality occured.

- Irritation:
Application of the test article to healthy intacct rabbit skin resulted in a primary irritation score of 0.0.
Local signs (mean values from 24 to 72 hours consisted of grade 0.0 erythema and grade 0.0 edema.
No sigs of irritation were observed.

- Coloration:
Black staining by the test article of the treated skin was observed.

- Corrosion:
No irreversible alterations of the treated skin were obbserved nor were corrosve effects evident on the skin.

- Body weight:
The body weight of all rabbits were cnsidered to be within the normal range of variability.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

FAT 41024/B is considered to be not irritating to rabbit skin.

Executive summary:

The purpose of this primary skin irritation study was to assess the possible irritation potential when single doses of the test article are placed on the skin of rabbits.

The test was performed according to the OECD Guideline 404 (Acute Dermal Irritation / Corrosion).

The primary skin irritation potential of the test article was investigated by topical application of 0.5 g to 6 cm2 intact dorsal skin of each of three young adult New Zealand White rabbits.

The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 ans 72 hours after removal of the dressing. The scores of each animal at the following reading times (24, 48, 72 hours) were used in calculating the respective mean values for each type of lesion.

The primary irritation score was calculated by totalling the individual cumulative scores at 24, 48 and 72 hours and then diving by the number of data points. The primary irritation score 0.0 (max. 8.0).

Local signs (mean values from 24 to 72 hours) consisted of grade 0.0 erythema and grade 0.0 edema.

Slight black and / or yellow staining of the treated skin was observed in all animals.

No corrosive effects were noted on the treated skin of any animal at any measuring interval.

The test article is considered to be "not irritating" to the rabbit skin.