N-nitro-N-(3-methyl-3,6-dihydro-2H-1,3,5-oxadiazin-4-yl)amine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August - September 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study; documentation sufficient for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

This study was performed in compliance with Good Laboratory Practice (GLP) in Switzerland, Procedures and Principles, March 1986. These procedures are based on the OECD Principles of GLP, adopted May 12, 1981 by Decision of the OECD Council C(81)30(Final)

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

431-060-1

EC Name:

-

Cas Number:

153719-38-1

Molecular formula:

C4H8N4O3

IUPAC Name:

3-methyl-N-nitro-3,6-dihydro-2H-1,3,5-oxadiazin-4-amine

Details on test material:

- Name of test material (as cited in study report): CA 2343 A
- Substance type: Intermediate of CGA 293343
- Physical state: powder
- Analytical purity: 96.7%
- Lot/batch No.: P.503005
- Storage condition of test material: 0-5°C
- Date of Reanalysis: December 1996
- Safety precautions: gloves and face masks

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Chemisch-pharmazeutische Fabrik, D-88397 Biberach
- Weight at study initiation: 2690 - 2970 g
- Housing: individually in metal cages
- Diet (e.g. ad libitum): ad libitum standard rabbit pellet (NAFAG 814, NAFAG, Gossau/SG, Switzerland)
- Water (e.g. ad libitum): ad libitum, water quality fulfilled the critical parameters in the specification of the "Schweizerisches Lebensmittelhandbuch"
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20°C +- 3°C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12 hours / 12 hours

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: untereated eye of the animals served as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml corresponding to about 57 mg

Duration of treatment / exposure:

one-time administration

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3

Details on study design:

SCORING SYSTEM: OECD guideline 405 - Scores for ocular lesions (see below) and
Classification of Irritant Potency (not shown) according to Annex iV, 3.2.6 of Council Directive 67/548/EEC (Commision Directive 93/21/EEC of April, 1993; Official Journal of the European Communities No. L 110 A, May 4, 1993).

Cornea
Opacity: degree of density (readings should be taken from most dense area)*
No ulceration or opacity.............................................................................................................................. 0
Scattered or diffuse areas of opacity (other than slight dulling of normal lustre); details of iris clearly visible ...................................................................................................................... 1
Easily discernible translucent area; details of iris slightly obscured ............................................................2
Nacrous area; no details of iris visible; size of pupil barely discernible ......................................................3
Opaque cornea; iris not discernible through the opacity ..............................................................................4
Maximum possible: 4
* The area of corneal opacity should be noted

Iris
Normal .........................................................................................................................................................0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia; or injection; iris reactive to light (a sluggish reaction is considered to be an effect ..................................1
Hemorrhage, gross destruction, or no reaction to light .................................................................................2
Maximum possible: 2

Conjunctivae
Redness (refers to palpebral and bulbar conjunctivae; excluding cornea and iris)
Normal .........................................................................................................................................................0
Some blood vessels hyperaemic (injected) ...................................................................................................1
Diffuse, crimson colour; individual vessels not easily discernible ...............................................................2
Diffuse beefy red..........................................................................................................................................3
Maximum possible: 3

Chemosis
Swelling (refers to lids and/or nictating membranes)
Normal .........................................................................................................................................................0
Some swelling above norma .........................................................................................................................1
Obvious swelling, with partial eversion of lids.............................................................................................2
Swelling, with lids about half closed ............................................................................................................3
Swelling, with lids more than half closed .....................................................................................................4
Maximum possible: 4

Results and discussion

In vivo

Irritant / corrosive response data:

see below in rich-text-field: "Remarks on results including tables and figures"
The mean scores of 24 through 72 hours after ocular administration indicated moderate circumscorneal hyperemia of the iris in normal swelling of eyelids inall rabbits. Diffuse areas of corneal opacity were noted in 2 of 3 rabbits. Observed eye reactions were completely reversible in all rabbits within 7 days.

Any other information on results incl. tables

All tables: First number given is always the control eye. The second number is the test eye.

Table 1: Irritant/corrosive response data for animal 1 (female) at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
60 min	0/1	0/1	0/2	0/2
24 h	0/1	0/1	0/2	0/1
48 h	0/0	0/1	0/1	0/1
72 h	0/0	0/0	0/1	0/0
Average 24h, 48h, 72h	0/0.33	0/0.67	0/1.33	0/.67
7 days	0/0	0/0	0/0	0/0

Table 2: Irritant/corrosive response data for animal 2 (male) at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
60 min	0/1	0/1	0/2	0/2
24 h	0/0	0/0	0/2	0/1
48 h	0/0	0/0	0/1	0/0
72 h	0/0	0/0	0/1	0/0
Average 24h, 48h, 72h	0/0	0/0	0/1.33	0/0.33
7 days	0/0	0/0	0/0	0/0

Table 3: Irritant/corrosive response data for animal 3 (male) at each observation time up to removal of each animal from the test

Score at time point / Reversibility	Cornea	Iris	Conjunctivae	Chemosis
	Max. score: 4	Max. score: 2	Max. score: 3	Max. score: 4
60 min	0/1	0/1	0/2	0/2
24 h	0/1	0/1	0/2	0/1
48 h	0/0	0/1	0/1	0/0
72 h	0/0	0/1	0/1	0/0
Average 24h, 48h, 72h	0/0.33	0/1	0/0.33	0/0.33
7 days	0/0	0/0	0/0	0/0

Table of body weights:

Animal No	678/female	007/male	509/male
At start of test	2740	2690	2970
After 3 days	2780	2670	3010
After 7 days (end)	2850	2910	3050

A transitory slight loss of body weight in rabbit No. 007 was recorded on experimental day 3.

Mortality was unaffected by the treatment.

Rabbit No. 007 vocalised upon instillation of the test article. Otherwise, thre were no remarkable in-life observations.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

When evaluated according to Council Directive 67/548/EEC, adapted to technical progress by Commission Directive 93/21/EEC oxadiazinamine has not to be classified as eye irritating.

Executive summary:

This study has been performed in compliance with Good Laboratory Practice (GLP) in . The study has been conducted according to OECD guideline 405 adopted February 24, 1987, by the OECD council, and on Annex V, part B of Council Directive 79/831/EEC (Commission Directive 92/69/EEC of July 31, 1992).

The test article (0.1 ml corresponding to 57 mg) was placed into the conjunctival sac of the left eye of each animal. Lids were then hold together for about 1 second. The untreated eye served as control.

The animals were checked daily for mortality and systemic symptoms. Body weights were recorded at start, after 3 days and on conclusion of the study on day 7. Ocular reactions were evaluated 1, 24, 48, 72 hours after instillation of the test article according to the OECD scoring system. Animals were observed for 7 days to determine the reversibility of the ocular reactions.

There was no mortality. One rabbit vocalised upon instillation of the test compound. No other systemic symptoms were recorded. In one animal, a transitory slight body weight loss was recorded on experimental day 3.

Grade 1 cornea opacity was seen in all test animals 1 hour after test article instillation, and in two animals 24 hours after instillation. Iris lesions were recorded in all test animals 1 hour after test article instillation in two animals 24 hours and 48 hours after instillation in one animal still existing after 72 hours. Conjunctival reactions (redness and/or chemosis) up to grade 2 were noted in all animals at all observation time pints until 72 hours of the test compound. The highest mean score of 24 to 72 hours after instillation was 1.33 for conjunctival redness, thus laying below the threshold of significance.

Observed ocular reactions were fully reversed in all animals by day 7.

When evaluated according to Council Directive 67/548/EEC, adapted to technical progress by Commission Directive 93/21/EEC, the substance oxadiazinamine has not to be classified as eye irritating.