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

Eye irritation

Administrative data

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes (incl. QA statement)

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:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Identification: FAT 41024/B
- Description: Black powder solid
- Batch number: ARL 83-PP 1/98 UL
- Expiry date: 31 March 2005
- Purity/Composition: 94%
- Stability of the test article: Stablle at storage conditions
- Storage conditions: At room temperature (ca. 20°C) away from direct sunlight
- Safety precautions: Gloves, goggles and face mask were sifficient to assure personnel health and safety

Test animals / tissue source

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.

Test system

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

Results and discussion

In vivo

Results

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.

Any other information on results incl. tables

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.

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 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.