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

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

August 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:

1996

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal 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: 93.7 %
- Purity test date:
- Lot/batch No.: P.503005
- Staorage conditions: 0 - 5°C
- Reanalysis date: December 1996
- Safety precautions: gloves and face mask

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Dr. K. Thomae GmbH, Chemisch-pharmazeutische Fabrik, D-88397 Biberach
- Age at study initiation: 2-3 month
- Weight at study initiation: 2700-2970 g
- Housing: individually in metal cages
- Diet (e.g. ad libitum): ad libitum standard rabbit pellét - NAFAG 814., NAFAG, Gossau/SG, Switzerland
- Water (e.g. ad libitum): ad libitum fresh water which fulfilled the critical parameters in the specifications of the "Schweizerisches Lebensmittelbuch" (Edition 1972)
- Acclimation period: at least 5 days


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

Test system

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

other: 0.5% (w/v) carboxymethylcelllose in 0.1% (w/v) aqueous polysorbate 80

Controls:

other: A control gauze patch was applied to the contralateral flank of each animal

Amount / concentration applied:

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

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5 % (w/v) carboxymethylcellulose in 0.1% (w/v) aqueous polyorbate 80

Duration of treatment / exposure:

The patches with the test substance were held in place for 4 hours by an adhesive tape.

Observation period:

The degree of skin reactions was evaluated 1, 24, 48, and 72 hours after removing the gauze patches according to the OECD scoring system. In case of persisting skin reactions on day 3 after gauze patch removal. the observation period may be extended.
Animals were observed for 7 days to determine the reversibility of the skin reactions.

Number of animals:

The test was performed on 3 male rabbits.

Details on study design:

TEST SITE
- Area of exposure: approx. 6 cm² of the right flank of each animal
- Type of wrap if used: adhesive tape (Isoplast, Isoplast AG, CH-5200 Brugg)

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 (beef redness) to eschar formation preventing grading of erythema.....................4
Maximum possible: 4

Oedema Formation
No oedema........................................................................................................................................................0
Very slight oedema (barely perceptible) ......................................................................................................1
Slight oedema (edges of area well defined by definite raising) ................................................................2
Moderate oedema (raised approximately 1 mm) ........................................................................................3
Severe oedema (raised more than 1 mm and extending beyond area of exposure) ............................4
Maximum possible: 4

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 1 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 24 h

Results 2

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 24 h, 48 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 3

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 1 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 24 h

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

other: 24 h, 48 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 5

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

other: 72 h

Remarks on result:

other: found dead due to a damaged kidney after an accident during examination

Results 6

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 1 h, 24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Results 7

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

other: 48 h, 72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 8

Irritation parameter:

edema score

Basis:

animal #2

Time point:

other: 1 h, 24 h, 48 h, 72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Results 9

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

other: 1 h, 24 h, 48 h, 72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 10

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 1 h, 24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 48 h

Results 11

Irritation parameter:

edema score

Basis:

animal #3

Time point:

other: 48 h, 72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Any other information on results incl. tables

The mean of 24 - 72 hours for

erythema scores:

animal 1 = 0 (found dead after 72 hours, therefore mean 24 - 48 h, see also remark above)

animal 2 = 0.33

animal 3 = 1

Edema scores:

animal 1 = 0 (found dead after 72 hours, therefore mean 24 - 48 h, see also remark above)

animal 2 = 0

animal 3 = 0.33

Observed skin reactions were fully reversed in surviving animals by day 7.

No erythema and edema were observed on the control flanks of each animal at all times

Body weights were unaffected by the treatment. There were no remarkable in –life observations.

Table of body weights: (all animals are male ones)

Animal No	639 (1)	643(2)	(3) 653
At start of test	2970	2890	2700
After 3 days	Ex. 2910	2960	2780
After 7 days	- - -	3010	2850

Applicant's summary and conclusion

Interpretation of results:

slightly irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

Very slight erythema was recorded in all rabbits 1 hour after gauze patch removal, in 2 of 3 animals after 24 hours and in 1 animal at up to 72 hours. Very slight edema was seen in 2 of 3 animals 1 hour after gauze patch removal and in 1 animal after 24 hours.
Observed skin reactinos were fully reversed in surviving animals by day 7 after removal of the gauze patches.

Executive summary:

This study has been performed in compliance with Good Laboratory Practice (GLP) in . The study has been conducted according to OECD guideline 404 adopted July 17, 1992, 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).

A gauze patch with 0.5 test material (oxadiazinamine), moistened with 0.5% (w/v) carboxymethylcellulose in 0.1% (w/v) aqueous polysorbate 80, was placed on the shaved skin of three male rabbits and loosely covered with aluminium foil for 4 hours. The controlateral flank was covered with a control patch moistened with 0.5% (w/v) carboxymethylcellulose in 0.1% (w/v) aqueous polysorbate 80 and with aluminium foil.

The animals were checked daily for mortality and systemic symptoms. Body weights were recorded at start, after three days and on conclusion of the study on day seven. Skin reactions were evaluated 1, 24, 48, and 72 hours after removing the gauze patches according to the OECD scoring system. Animals were observed for 7 days to determine the reversibility of the skin reactions.

One animal was found dead on day 3 due to a damaged kidney after an accident during examination. No systemic symptoms were recorded in any of the test animals. Body weight development was within a normal range.

Grade 1 erythema and/or oedema were seen in the test animals 1 hour and 24 hours after gauze patch removal, erythema still existing existing in one animal after 72 hours.

Observed skin reactions were fully reversed in surviving animals by day 7.

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 skin irritating.