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

Endpoint summary

Description of key information

- In vivo Primary skin irritation study in rabbits (4-hour semi-occlusive application): Negative.
- In vivo Primary eye irritation study in rabbits: Negative.

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:

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.

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

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

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

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)

Irritation parameter:

erythema score

Basis:

mean

Remarks:

1 male and 2 females

Time point:

other: 1-72 hours

Score:

0

Irritation parameter:

edema score

Basis:

mean

Remarks:

1 male and 2 females

Time point:

other: 1-72 hours

Score:

0

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.

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.

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:

14 October 1998 to 22 October 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP methodology followed and OECD Guideline 405 used to performed the experiment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues 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°62/98)
- Water: Community tap water from Itingen, 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. Recorded music was played for approximately 8 hours during the light period.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

Solid: 0.1 gr/animal (left eye only)

Duration of treatment / exposure:

One application

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 rabbits (1 male and 2 females)

Details on study design:

SCORING SYSTEM:
Grading of Ocular lesions: Opacity of density (area most dense taken for reading).
CORNEA
- No ulceration or opacitiy: 0
- Scattered or diffuse areas of opacity (other than slight dulling of normal luster), details od iris clearly visible: 1
- Easily discernible translucent area, details of iris slightly obscured: 2
- Nacreous area, no details of iris visible, size of pupil barely discernible: 3
- Opaque cornea, iris not discernible through the opacity: 4

IRIS
- Normal: 0
- Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperemia, or injection, any 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

CONJUNCTIVAE
Redness (refers to most severe reading of palepbral and bulbar conjunctivae when compared with control eye
- Blood vessels normal: 0
- Some blood vessels definitely hyperemic (injected): 1
- Diffuse, crimson, individual vessels not easily descernible: 2
- Diffuse red: 3
Chemosis: lids and/or nictitating membranes
- No swelling:0
- Any swelling above normal (including nictitating mambranes): 1
- Obvious swelling partiel eversion of lids: 2
- Swelling with lids about half-closed: 3
- Swelling with lids more than half-closed: 4

Irritation parameter:

other: Primary eye irritation score

Basis:

mean

Remarks:

1 male and 2 females

Time point:

other: 24-72 hours

Score:

0

Max. score:

13

Irritant / corrosive response data:

- IRRITATION:
Application of the test article to healthy rabbit conjunctivae resulted in a primary irritation score of 0.00. In all animals, hyperemia of the scleral blood vessels and slight to moderate watery discharge was observed. In two animals, slight to moderate swelling of the conjunctivae including the nictitating membrane was noted after one hour. All signs of irritation were reversible after 72 hours.

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.

Coloration:

- Reversible blue staining of the cornea, conjunctivae, the sclera and the lid hairs by the test article was observed.

Body weight:

The body weight of all rabbits were considered to be within the normal range of variability.

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 the rabbit eye.

Executive summary:

This experiment was performed according to the OECD Guideline 405 (Acute Eye Irritation / Corrosion).

The primary irritation potential of the test article was investigated by instillation of 0.1 g into one eye of each of three younf adult New Zaeland White rabbits. The treated eyes were not rinsed after application.

Scoring of irritation effects was performed approximately 1,24, 48 and 72 hours after test article application. The scores of each animal at the following reading times (24, 48 ans 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 dividing the resulting total by the numebr of dara points. The primary irritation score was 0.00 (max 13).

In all animals hyperemia of the scleral blood vessels and watery discharge was observed. In two animals swelling of the conjunctivae

including the nictitating membrane was noted after one hour. All signs of irritation were reversible after 72 hours.

Reversible blue staining of the cornea, the conjunctivae, the sclera and the lid hairs of the treated eyes by the test article was observed in all animals.

No corrosion was observed at any of the measuring intervals.

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

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The in vivo primary skin irritation experiment in rabbits was performed on one male and 2 females. FAT 41024/B was applied to the intact skin during 4 hours at an amount of 0.5 g per animal.

Application of the test article to healthy skin resulted in a primary score of 0.00. Local signs (values from 24 to 72 hours) consisted of grade 0.00 erythema and grade 0.00 edema. No signs of irritation were observed.

The in vivo primary eye irritation experiment in rabbits was performed on one male and two females. An amount of 0.1 g of FAT 41024/B was applied to left eye of each rabbit. No clinical signs of systemic toxicity were observed and no mortality occured. No corrosion of the cornea was observed at any of the reading times.

Application of the test article to healthy rabbit conjunctivae resulted in a primary irritation score of 0.00. In all animals, hyperemia of the scleral blood vessels and slight to moderate watery discharge was observed. In two animals, slight to moderate swelling of the conjunctivae including the nictitating membrane was noted after one hour. All signs of irritation were reversible after 72 hours.

Justification for selection of skin irritation / corrosion endpoint:
Only this study is available

Justification for selection of eye irritation endpoint:
Only this study is available

Justification for classification or non-classification

Based on the above mentioned results the substance does not need to be classified according to CLP regulation (Regulation EC No.1272/2008) and DSD (Directive 67/548/EEC).