N-(butoxymethyl)acrylamide

Endpoint summary

Description of key information

The skin irritation potential of the test material was assessed in an in vivo study using a method similar to OECD 404 using three New Zealand white rabbits. The substance is not classified as a skin irritant.

 

The eye irritation potential of the test material was assessed in an in vivo study using a method similar to OECD 405 using three New Zealand white rabbits. The substance is not classified as an eye irritant.

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:

26 June 2006 to 13 October 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

the test material was applied to abraded as well as intact skin

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: The pre-test body weight range was 2.6 - 3.0 kg
- Age at study initiation: Approximately 14 weeks
- Housing: The animals were housed 1 per cage in suspended cages. Bedding was placed beneath the cages and changed at least three times per week.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Equilibrated for at least three days

ENVIRONMENTAL CONDITIONS
- Temperature: Not reported, the room was temperature controlled
- Photoperiod: 12 hour light / dark cycle

IN-LIFE DATES:
- From: 07 June 2006
- To: 03 July 2006

Type of coverage:

occlusive

Preparation of test site:

other: Rabbits were clipped and the substance applied to both intact and abraded skin

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.5 mL to two areas (one intact and one abraded) for a total dermal exposure dose of 1.0 mL

VEHICLE
The test material was used as received

OTHER
Rabbits were examined and found to be free of dermal irritation and skin abnormalities prior to the start of the test. The left side of each animal was abraded with a bent tip needle. Three abrasions approximately 2 - 3 cm apart, extending the length of the exposure site were made. The abrasions were sufficiently deep to penetrate the stratum corneum but not deep enough to produce bleeding. The right side of each animal remained intact.

Duration of treatment / exposure:

4 hours

Observation period:

7 days

Number of animals:

3 rabbits; one male and two female.

Details on study design:

TEST SITE
- Area of exposure: Back and sides of each animal.
- Type of wrap if used: 2.5 x 2.5 cm 4 ply surgical gauze patches which were secured with non-irritating adhesive tape. The torso was wrapped with plastic which was secured with non-irritating adhesive tape. In order to permit dermal observations during the 4-hour exposure period, the wrappings were fitted with a window which was opened to examine the test site. The sites were semi-occluded for 4 hours at which time the patches were removed.

REMOVAL OF TEST SUBSTANCE
- Washing: Not specified

OBSERVATION TIME POINTS
Animals were observed for skin reactions, including ulceration and necrosis, during the 4 hour exposure period at 3 minutes post-dose and at 1 and 4 hours post-dose. The test sites were scored again at 24, 48 and 72 hours and again on day 7. Erythema and eschar were scored according to the numerical Draize scale. Additional signs were described.
Bodyweights were recorded pre-test. The general health of each animal was monitored at each observation time. All animals were humanely sacrificed using CO2 following study termination.

SCORING SYSTEM:
Erythema and eschar formation:
0: No erythema
1: Very slight erythema (barely perceptible)
2: Well-defined erythema
3: Moderate to severe erythema
4: Severe erythema (beet redness) to slight eschar formation (injuries in depth)

Oedema formation:
0: No oedema
1: Very slight oedema (barely perceptible)
2: Slight oedema (edges of area well-defined by definite raising)
3: Moderate oedema (edges raised approximately 1 millimetre)
4: Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure)

Irritation parameter:

erythema score

Basis:

animal #1

Remarks:

G8728 Male

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Results from intact skin only.

Irritation parameter:

erythema score

Basis:

animal #2

Remarks:

G8760 Female

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: Results from intact skin only.

Irritation parameter:

erythema score

Basis:

animal #3

Remarks:

G8761 Female

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Remarks on result:

other: Results from intact skin only.

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

G8728 Male

Time point:

24/48/72 h

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: Results from intact skin only.

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

G8760 Female

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: Results from intact skin only.

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

G8761 Female

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable - No effects observed

Remarks on result:

other: Results from intact skin only

Irritant / corrosive response data:

Erythema was absent at three minutes after dosing, absent to very slight at one hour after dosing and very slight at four hours after dosing. Oedema was absent at three minutes and one hour after dosing and absent to slight at four hours after dosing.
At 24, 48 and 72 hours after patch removal, erythema was absent to well defined. Oedema was absent to very slight at 24 hours after patch removal and absent to well-defined at 48 and 72 hours.
Both erythema and oedema were absent by Day 7.

Other effects:

One instance of diarrhoea was the only abnormal physical sign noted during the observation period.

Table 1: Erythema and Eschar Formation

Test Condition	Rabbit No. / Sex	G8728/M	G8760/F	G8761/F
	Observation Time
Intact skin	3 minutes"	0	0	0
	1 hour"	0	0	0
	4 hours"	1	1	1
	24 hours**	1	1	1
	48 hours**	2	0	1
	72 hours**	2	0	0
	7 days**	0*	0*	0*
Abraded skin	3 minutes	0	0	0
	1 hour	1	0	0
	4 hours	1	1	1
	24 hours	2	1	0
	48 hours	2	0	0
	72 hours	2	0	0
	7 days	0*	0*	0*

" Post dose observation

* Reclipped

** Post-patch removal observation

Table 2: Oedema

Test Condition	Rabbit No. / Sex	G8728/M	G8760/F	G8761/F
	Observation Time
Intact skin	3 minutes	0	0	0
	1 hour	0	0	0
	4 hours	1	0	0
	24 hours	1	1	1
	48 hours	2	0	0
	72 hours	2	0	0
	7 days	0	0	0
Abraded skin	3 minutes	0	0	0
	1 hour	0	0	0
	4 hours	1	0	1
	24 hours	1	1	0
	48 hours	2	0	0
	72 hours	2	0	0
	7 days	0	0	0

Interpretation of results:

other: Not classified in accordance with EU criteria

Conclusions:

The test material was not irritating to the 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:

26 June 2006 to 13 October 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Instead of one eye remaining untreated and serving as control, both eyes were treated and one was rinsed.

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: The pre-test body weight range was 2.6 - 3.0 kg
- Age at study initiation: Approximately 14 weeks
- Housing: The animals were housed 1 per cage in suspended cages. Bedding was placed beneath the cages and changed at least three times per week.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Equilibrated for at least three days

ENVIRONMENTAL CONDITIONS
- Temperature: Not reported, the room was temperature controlled
- Photoperiod: 12 hour light / dark cycle

IN-LIFE DATES:
- From: 07 June 2006
- To: 03 July 2006

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Initially, the right eye of one rabbit was dosed. Since no discomfort was noted at the time of dosing, the left eye and both eyes of the remaining two rabbits were dosed. The test article was placed by syringe into the conjunctival sac which was formed by gently pulling the lower eyelid away from the eye. After instillation, the lids were held together briefly to ensure adequate distribution of the test article.

VEHICLE
The test material was used as supplied.

OTHER
Prior to the start of the test, both eyes of each animal were examined for any evidence of irritation or abnormalities of the cornea, iris and/or conjunctiva.

Duration of treatment / exposure:

- Left eye: 20-30 seconds prior to washing
- Right eye: Test material was not removed

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3 rabbits; one male and two female.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing: The left eye of each rabbit was flushed for one minute with lukewarm water. The right eye of each animal remained unwashed.
- Time after start of exposure: Twenty to thirty seconds after instillation of the test article.

SCORING SYSTEM:
Ocular reactions were graded according to the numerical Draize technique. Both eyes of each rabbit were examined for irritation of the cornea, iris and conjunctiva at one hour post-dose, and at 24, 48 and 72 hours post-dose. The eyes were scored again on Day 7. Additional signs were described.

CORNEA
Opacity: degree of density (area most dense is taken for reading).
0: No opacity
1: Scattered or diffuse areas of opacity, details of iris clearly visible
2: Easily discernible translucent area, details of iris slightly obscured
3: Opalescent area, no details of iris visible, size of pupil barely discernible
4: Opaque, iris invisible

IRIS
0: Normal
1: Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination of any thereof) still reacting to light (sluggish reaction is positive)
2: No reaction to light, haemorrhage, gross destruction (any of all these or all together)

CONJUNCTIVAE
Redness: (refers to the most severe effect of palpebral and bulbar conjunctivae excluding cornea and iris)
0: Vessels normal
1: Vessels definitely injected above normal
2: More diffuse, deeper crimson red, individual vessels not easily discernible
3: Diffuse beefy red
Chemosis
0: No swelling
1: Any swelling above normal (includes nictating membranes)
2: Obvious swelling with partial eversion of lids.
3: Swelling with lids about half closed
4: Swelling with lids about half closed to completely closed
Discharge
0: No discharge
1: Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)
2: Discharge with moistening of the lids and hairs just adjacent to lids
3: Discharge with moistening of the lids and hairs a considerable area around the eye

TOOL USED TO ASSESS SCORE:
A Mini Maglite® flashlight equipped with a high intensity bulb was used to aid in the examination.

OTHER
Body weights were recorded pre-test. The general health of the animals was monitored at each observation time. All animals were humanely scarified using CO2 following study termination.

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of all three animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: Not applicable - no effects observed

Remarks on result:

other: Based on unwashed eyes only.

Irritation parameter:

iris score

Basis:

mean

Remarks:

of all three animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: Not applicable - no effects observed

Remarks on result:

other: Based on unwashed eyes only.

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

of all three animals

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: Not applicable - no effects observed

Remarks on result:

other: Based on unwashed eyes only.

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of all three animals

Time point:

24/48/72 h

Score:

0.11

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Remarks on result:

other: Based on unwashed eyes only.

Irritation parameter:

other: discharge

Basis:

mean

Remarks:

of all three animals

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

other: Not applicable - no effects observed

Remarks on result:

other: Based on unwashed eyes only.

Irritant / corrosive response data:

- Unwashed eyes: There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation was noted in 3/3 eyes which cleared within 48 hours.
- Washed eyes: There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation was noted in 3/3 eyes which cleared by Day 7.

Other effects:

One instance of diarrhoea was the only abnormal physical sign noted during the observation period.

Table 1: Ocular Findings in Right Eye - Unwashed

Animal No.	Tissue	Reading	1 h	24 h	48 h	72 h	7 Days
G8728 M	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0	0
	Iris		0	0	0	0	0
	Conjunctiva	Redness	1	0	0	0	0
		Chemosis	0	0	0	0	0
G8760 F	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0	0
	Iris		0	0	0	0	0
	Conjunctiva	Redness	1	0	0	0	0
		Chemosis	2	0	0	0	0
G8761 F	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0	0
	Iris		0	0	0	0	0
	Conjunctiva	Redness	1	0	0	0	0
		Chemosis	2	1	0	0	0

 

Table 2: Ocular Findings in Left Eye - Washed

Animal No.	Tissue	Reading	1 h	24 h	48 h	72 h	7 Days
G8728 M	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0	0
	Iris		0	0	0	0	0
	Conjunctiva	Redness	1	0	0	0	0
		Chemosis	0	0	0	0	0
G8760 F	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0	0
	Iris		0	0	0	0	0
	Conjunctiva	Redness	1	2	2	1	0
		Chemosis	1	2	1	0	0
G8761 F	Cornea	Opacity	0	0	0	0	0
		Area	0	0	0	0	0
	Iris		0	0	0	0	0
	Conjunctiva	Redness	1	2	1	1	0
		Chemosis	1	1	1	0	0

 

Interpretation of results:

other: Not classified in accordance with EU criteria

Conclusions:

The test material is not irritating to eyes.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

The skin irritation potential of the test material was assessed in an in vivo study using a method similar to OECD 404 using three New Zealand white rabbits. 

Two aliquots of the test material totalling 1.0 mL were applied to the clipped skin on the back and side of three animals and covered with an occlusive dressing. Animals were observed for skin reactions, including ulceration and necrosis, during the 4 hour exposure period at 3 minutes post-dose, and at 1 and 4 hours post-dose. The test sites were scored again at 24, 48 and 72 hours and again on day 7. Bodyweights were recorded pre-test. The general health of each animal was monitored at each observation time. In order to permit dermal observations during the 4-hour exposure period, the wrappings were fitted with a window which was opened to examine the test site. The sites were semi-occluded for 4 hours at which time the patches were removed.

The mean erythema score at 24, 48 and 72 hours was recorded for each rabbit and ranged from 0.3 to 1.67 out of a maximum score of 4.

The mean oedema score at 24, 48 and 72 hours was recorded for each rabbit and ranged from 0.0 to 1.67 out of a maximum score of 4.

Therefore, while slight erythema and oedema were recorded initially, the reactions were not sufficient to classify the substance as a skin irritant. Both erythema and oedema were absent by day 7.

 

Eye irritation

The eye irritation potential of the test material was assessed in an in vivo study using a method similar to OECD 405 using three New Zealand white rabbits. 

Initially, the right eye of one rabbit was dosed with 0.1 mL. Since no discomfort was noted at the time of dosing, the left eye and both eyes of the remaining two rabbits were dosed. The test article was placed by syringe into the conjunctival sac which was formed by gently pulling the lower eyelid away from the eye. After instillation, the lids were held together briefly to ensure adequate distribution of the test article. Prior to the start of the test, both eyes of each animal were examined for any evidence of irritation or abnormalities of the cornea, iris and/or conjunctiva. Both eyes of each rabbit were examined for irritation of the cornea, iris and conjunctiva at one hour post-dose, and at 24, 48 and 72 hours post-dose. The eyes were scored again on Day 7. 

The left eye of each rabbit was flushed for one minute with lukewarm water twenty to thirty seconds after instillation of the test article. The right eye of each animal remained unwashed.

Washed eyes: There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation was noted in 3/3 eyes which cleared by Day 7.

Unwashed eyes: There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation was noted in 3/3 eyes which cleared within 48 hours. The mean chemosis score at 24, 48 and 72 hours was recorded for each rabbit and ranged from 0.0 to 0.33 out of a maximum score of 4.

The reactions were not sufficient to classify the substance as an eye irritant.

Justification for classification or non-classification

Skin irritation

The mean erythema and oedema scores at 24, 48 and 72 hours were recorded for each rabbit and ranged from 0.3 to 1.67 and 0.0 to 1.67, respectively, out of a maximum score of 4.

Therefore, while slight erythema and oedema were recorded initially, the reactions were not sufficient to classify the substance as a skin irritant.

 

Eye irritation

There was no corneal opacity or iritis noted at any observation period. Conjunctival irritation was noted in 3/3 eyes which cleared within 48 hours. The mean chemosis score at 24, 48 and 72 hours was recorded for each rabbit and ranged from 0.0 to 0.33 out of a maximum score of 4.The reactions were not sufficient to classify the substance as an eye irritant.