1,4-diazabicyclooctane

Endpoint summary

Description of key information

1,4-Diazabicyclo[2.2.2]octane is severely irritating to the eye of rabbits and is moderately irritating to the skin of rabbits upon direct contact. 1,4-Diazabicyclo[2.2.2]octane vapors produce transitory irritation of the eyes and mucous membranes.

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study pre-dates GLPs Test material purity was not documented.

Qualifier:

according to guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: Section 1500.41 - Federal Hazardous Substances A c t Regulations - 16 CFR - Po 124.

Principles of method if other than guideline:

Draize Test
A group of six albino rabbits was clipped over a wide area. One side of the animals' backs was abraded a t one site with a
lancet sufficiently deep to penetrate the stratum corneum but not enter the derma t o produce bleeding. The skin of the
other side was allowed to remain intact.
A 0.5 gm portion of material was applied to an abraded and an intact skin site on the same rabbit. Gauze patches were then
placed over the treated areas and an impervious material was wrapped snugly around the trunks of the animals to hold the
patches in place.
The wrapping was removed at the end of the twenty-four hour period and the treated areas were examined. Readings were
also made after seventy-two hours. The Draize method of scoring was employed.

GLP compliance:

no

Species:

rabbit

Type of coverage:

occlusive

Preparation of test site:

other: clipped+abradded

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

undiluted

Duration of treatment / exposure:

24 hour(s)

Observation period:

72 h

Number of animals:

6

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24h and 72h

Score:

2.92

Reversibility:

not specified

Remarks on result:

other: Study was done before OECD guidelines were established, hence there are no 48 h data available.

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

intact skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

intact skin

Time point:

72 h

Score:

3

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

abraded skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

abraded skin

Time point:

72 h

Score:

3

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

intact skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Remarks on result:

other:

Remarks:

erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

intact skin

Time point:

72 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

abraded skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

abraded skin

Time point:

72 h

Score:

4

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

intact skin

Time point:

72 h

Score:

0

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

abraded skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

abraded skin

Time point:

72 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #4

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #4

Remarks:

intact skin

Time point:

72 h

Score:

0

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #4

Remarks:

abraded skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #4

Remarks:

abraded skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #5

Remarks:

intact skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #5

Remarks:

intact skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #5

Remarks:

abraded skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #5

Remarks:

abraded skin

Time point:

72 h

Score:

2

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #6

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #6

Remarks:

intact skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #6

Remarks:

abraded skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal #6

Remarks:

abraded skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

erythema score

Basis:

animal: all animals

Remarks:

intactact and abraded skin average subtotal

Time point:

24/48/72 h

Score:

6.51

Reversibility:

not specified

Remarks on result:

other: erythema and eschar formation

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

intact skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

intact skin

Time point:

72 h

Score:

3

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

abraded skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

abraded skin

Time point:

72 h

Score:

3

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

intact skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

abraded skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

abraded skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

intact skin

Time point:

72 h

Score:

0

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

abraded skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

abraded skin

Time point:

72 h

Score:

2

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #4

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #4

Remarks:

intact skin

Time point:

72 h

Score:

0

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #4

Remarks:

abraded skin

Time point:

24 h

Score:

2

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #4

Remarks:

abraded skin

Time point:

72 h

Score:

2

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #5

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #5

Remarks:

intact skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #5

Remarks:

abraded skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #5

Remarks:

abraded skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #6

Remarks:

intact skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #6

Remarks:

intact skin

Time point:

72 h

Score:

0

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #6

Remarks:

abraded skin

Time point:

24 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Basis:

animal #6

Remarks:

abraded skin

Time point:

72 h

Score:

1

Reversibility:

not specified

Irritation parameter:

edema score

Remarks:

subtotal

Basis:

mean

Time point:

other: 24 h and 72 h

Score:

5.16

Reversibility:

not specified

Remarks on result:

other: Study was done before OECD guidelines were established, hence there are no 48 h data available.

Irritation parameter:

other: erythema and eschar formation

Remarks:

subtotal

Basis:

mean

Time point:

other: 24 h and 72 h

Score:

6.51

Reversibility:

not specified

Remarks on result:

other: Study was done before OECD guidelines were established, hence there are no 48 h data available.

Irritation parameter:

edema score

Basis:

animal #1

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #2

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #3

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #4

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #5

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

edema score

Basis:

animal #6

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #4

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #5

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Irritation parameter:

erythema score

Basis:

animal #6

Time point:

48 h

Reversibility:

not specified

Remarks on result:

not measured/tested

Barely perceptible to severe erythema and barely perceptible
to moderate edema were observed.  A primary irritation index
of 2.92 was calculated.

Interpretation of results:

irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The material is i r r i t a t i n g t o the skin of albino rabbits.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP study

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

(obersvation duration 7 days instead of 21 days)

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Kleintierfarm Madoer in 4414 Fuellinsdorf / Switzerland
- Age at study initiation: 13 - 14 weeks
- Weight at study initiation: 2.2 - 2.4 kg
- Housing: single
- Diet: Pelleted standard Kliba 341, Batch 21/86 rabbit maintenance diet ("Kliba", Klingentalmuehle AG, 4303 Kaiseraugst, Switzerland); ad libitum.
- Water: Community tap water from Itingen; ad libitum.
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3
- Humidity (%): 40 - 70
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: The right eye remained untreated and served as the reference control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 g per animal was placed in the conjunctival sac of the left eye of each animal after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test article.

Duration of treatment / exposure:

One single treatment, the eye was not rinsed.

Observation period (in vivo):

7 days (reading recorded at 1 h, 24 h, 48 h, 72 h and 7 d after application)

Number of animals or in vitro replicates:

2 males, 1 female

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: no

SCORING SYSTEM: Draize

TOOL USED TO ASSESS SCORE: hand-slit lamp
Eye examinations were made with a slit-lamp 30 SL (C. Zeiss AS, Zuerich / Switzerland) and a Varta Cliptrix diagnostic-lamp (A. Riegger, Basel / Switzerland).

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.3

Max. score:

4

Reversibility:

not reversible

Remarks:

within 7 days

Remarks on result:

other: score 2 after 7 days.

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: score 2 after 7 days

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: score 2 after 7 days

Irritation parameter:

iris score

Basis:

mean

Remarks:

(3 animals)

Time point:

24/48/72 h

Score:

1

Max. score:

2

Reversibility:

not reversible

Remarks:

within 7 days

Remarks on result:

other: Score 1 in 2/3 animals and score 0 in 1/3 animals after 7 days

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

(3 animals)

Time point:

24/48/72 h

Score:

3

Max. score:

3

Reversibility:

not reversible

Remarks:

within 7 days

Remarks on result:

other: Score 3 in 2/3 animals; Score 2 in 1/3 animals after 7 days.

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.7

Max. score:

4

Reversibility:

not fully reversible within: within 7 days

Remarks on result:

other: score 2 after 7 days

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

not fully reversible within: 7 days

Remarks on result:

other: score 1 after 7 days

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Corneal opacity and partly deformation of the cornea was observed in all rabbits within 1 hour to 7 days after treatment.

Other effects:

No acute toxic symptoms were observed in the animals during the test period, and no mortality occurred.

Exposition: Single application to the conjunctival sac of the eyelid; the substance is not washed out
Animal	Reading	Cornea	Iris	Conjunctiva		Additional findings
		Opacity		Redness	Chemosis
1	1 h	1	0	3	2	Cornea: opacity, whole area; Conjunctivae: erythema, generally severe; edema, clearly visible; including nictating membrane; Sclera: not visible; Discharge: severe, lidhair, slight wet.
2	1 h	1	0	2	2	Cornea: opacity, generally; Conjunctivae : erythema, edema, generally, clearly visible, including nictating membrane; Sclera: not visible; Discharge: severe, lidhair, severe wet.
3	1 h	1	0	3	2	Cornea: opacity, whole area; Conjunctivae: erythema, generally severe; edema, clearly visible; including nictating membrane; Sclera: not visible; Discharge: severe, lidhair, severe wet.
1	24 h	1	1	3	2	Cornea: opacity, whole area; Conjunctivae: erythema, generally severe; edema, clearly visible; including nictating membrane; Sclera: erythema, vessels, dorsal; Discharge: severe, lidhair, severe wet; Iris: erythema, slight.
2	24 h	1	1	3	2	Cornea: opacity, whole area, ventral slight deformed; Conjunctivae: erythema, severe, ventral; edema, clearly visible, including nictating membrane; Sclera: erythema, generally; Discharge: severe, lidhair, severe wet; Iris: erythema.
3	24 h	1	1	3	2	Cornea: opacity, whole area; Conjunctivae: erythema, generally, severe; edema, clearly visible, including nictating membrane; Sclera: not visible; Discharge: severe, lidhair, severe wet; Iris: erythema.
1	48 h	1	1	3	3	Cornea: opacity, whole area; Conjunctivae: erythema, generally, several edema, severe; Sclera: not visible; Discharge: severe, eyes closed, adhered; Iris: erythema, slight .
2	48 h	1	1	3	2	Cornea: opacity, whole area, ventral slight deformed; Conjunctivae: erythema, severe, ventral; edema, clearly visible, including nictating membrane; Sclera: erythema, generally; Discharge: severe, lidhair, severe wet; Iris: erythema.
3	48 h	2	1	3	2	Cornea: opacity, severe, whole area; Conjunctivae: erythema, generally, severe; edema, clearly visible, including nictating membrane; Sclera: not visible; Discharge: severe, lidhair, severe wet; Iris: erythema.
1	72 h	2	1	3	3	Cornea: opacity, severe, iris, partly slight darkened; Conjunctivae: erythema, generally severe; edema, severe; Sclera: not visible; Discharge: severe, eyes closed, adhered; Iris: erythema.
2	72 h	1	1	3	2	Cornea: opacity, whole area, ventral slight deformed; Conjunctivae: erythema, severe, ventral; edema, clearly visible, including nictating membrane; Sclera: erythema, whole area; Discharge: severe, lidhair, severe wet; Iris: erythema.
3	72 h	2	1	3	2	Cornea: opacity, severe, whole area; Conjunctivae: erythema, generally, severe; edema, clearly visible, including nictating membrane; Sclera: not visible; Discharge: severe, lidhair, slight wet; Iris: erythema.
1	7 days	2	1	3	2	Cornea: opacity, severe, iris, partly slight darkened; Conjunctivae: erythema, generally, severe; edema, clearly visible, including nictating membrane; Sclera: erythema, severe, whole area; Discharge: severe, eyes closed, adhered; Iris: erythema.
2	7 days	2	1	3	1	Cornea: opacity, severe ventral; ventral, clearly visible deformed; Conjunctivae: erythema, generally, severe; edema, slight; Sclera: erythema, whole area; Discharge: slight, lidhair, dry; Iris: erythema, severe, slight reaction to light.
3	7 days	2	0	2	0	Cornea: opacity, severe ventral; Conjunctivae: erythema, ventral clearly visible; Sclera: erythema, slight ventral;
mean animal 1	24 - 72 h	1.33	1.00	3.00	2.67
mean animal 2	24 - 72 h	1.00	1.00	3.00	2.00
mean animal 3	24 - 72 h	1.67	1.00	3.00	2.00
mean	24 - 72 h total	1.25	0.75	3.00	2.22

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Migrated information Criteria used for interpretation of results: other: EU (CLP)

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Additional information

In two non-guideline Draize studies, 1,4-Diazabicyclo[2.2.2]octane was moderately irritating to the skin of rabbits following occlusive, 24-hour applications. In the one study, 0.5 g of crystalline material was applied. In the other study, 0.5 ml of a 50% solution was applied. Primary irritation indices of 2.92 and 4.05 were calculated, respectively.

In a guideline (OECD 405) study, 1,4-Diazabicyclo[2.2.2]octane was severely irritating to the eyes of rabbits. Corneal opacity and conjunctival irritation were the most notable findings. A mean primary irritation score of 7.1 out of 13 was calculated.

Rats exposed to a saturated vapor of 1,4-Diazabicyclo[2.2.2]octane for 8 hours exhibited transitory irritation of the eyes and mucous membranes

Justification for selection of skin irritation / corrosion endpoint:
Most recent and reliable study. It is done on the undiluted material so it is considered decisive for this endpoint.

Justification for selection of eye irritation endpoint:
Most recent and reliable study.

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: corrosive

Justification for classification or non-classification

Data is sufficient for classifiying 1,4-Diazabicyclo[2.2.2]octane as a skin irritant and as damaging to the eye.