N-[3-(trimethoxysilyl)propyl]aniline

Endpoint summary

Description of key information

skin irritation (OECD 402): not irritating
eye irritation (OECD 405): not irritating

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD 402 (Acute dermal toxicity)

Deviations:

no

GLP compliance:

yes

Species:

rat

Strain:

other: Crl:CD BR

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc., Portage, MI, USA
- Age at study initiation: young adult
- Weight at study initiation: 220-267 g
- Fasting: 18-20 h prior to dosing until 3-4 after dosing
- Housing: individual housing in mesh-bottom cages
- Diet: Purina Certified Rodent Chow #5002, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.1±0.2
- Humidity (%): 39.1-44.0
- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw (1.97 mL/kg)

Duration of treatment / exposure:

24 h

Observation period:

14 d

Number of animals:

5 animals per sex

Details on study design:

TEST SITE
- % coverage: 14-18% of total body surface
- Type of wrap if used: gauze bandaging secured with non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): application sites were wiped with disposable paper towels moistened with deionized water
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw, 1.92 mL/kg

SCORING SYSTEM: Draize scoring system

OTHER OBSERVATIONS:
- Frequency of observations and weighing: The rats were observed 1, 3, and 4 h post-dose on day of appplication (day 0) and once daily (clinical observation) or twice daily (mortality) thereafter. Body weights were obtained on the day of treatment and on day 7 and 14.
- Necropsy of survivors performed: yes (major organ systems of the cranial, thoracic and abdominal cavities were examined for all animals)
- Dermal observations: application sites were examined for erythema, oedema and other dermal findings 30-60 minutes after bandage removal and daily thereafter for thirteen days. The rats were clipped to facilitate dermal observations on study days 7 and 14.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

out of all 10 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Remarks on result:

other: Desquamation was observed in 6/10 animals (fully reversible in 4/6 animals). Eschar was observed in 1/10 animals (day 11-14). Erythema (grade 4) and edema (grade 1) were observed in 1 animal at the end of the study.

Irritation parameter:

edema score

Basis:

mean

Remarks:

out of all 5 males

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

The test material induced very slight to severe erythema, very slight edema and eschar on one animal. Desquamation was present on six animals. There were no other dermal findings. With the exception of one male and female with severe erythema, slight edema, eschar and/or desquamation, all dermal irritation completely subsided by day 11 or earlier.

Table 2: Dermal observations of the males in the acute dermal toxicity study.

Rat No. (males)	1		2		3		4		5
Observation time [day no]	Erythema	Edema	Erythema	Edema	Erythema	Edema	Erythema	Edema	Erythema	Edema
1	0d	0	0d	0	0	0	0	0	0	0
2	0d	0	0	0	0	0	0	0	0	0
3	0d	0	0	0	0	0	0	0	0	0
4	0d	0	0d	0	0	0	0	0	0d	0
5	0d	0	0d	0	0	0	0	0	0d	0
6	0d	0	0	0	0	0	0	0	0	0
7	0d	0	0	0	0d	0	0	0	0	0
8	0d	0	0	0	0d	0	0	0	0	0
9	0d	0	0	0	0d	0	0	0	0	0
10	0d	0	0	0	0d	0	0	0	0	0
11	0d	0	0	0	0	0	0	0	0	0
12	0d	0	0	0	0	0	0	0	0	0
13	0d	0	0	0	0	0	0	0	0	0
14	0d	0	0	0	0	0	0	0	0	0

d: desquamation

Table 3: Dermal observations of the females in the acute dermal toxicity study.

Rat No. (females)	1		2		3		4		5
Observation time [day no]	Erythema	Edema	Erythema	Edema	Erythema	Edema	Erythema	Edema	Erythema	Edema
1	0	0	0	0	0	0	0	0	0	0
2	0	0	0	0	0	0	0	0	0	0
3	0	0	0d	0	0	0	0	0	0	0
4	0	0	0d	0	0	0	0	0	0d	0
5	0	0	1d	0	0	0	0	0	0	0
6	0	0	1d	0	0	0	0	0	0	0
7	0	0	2d	0	0	0	0	0	0	0
8	0	0	2d	0	0	0	0	0	0	0
9	0	0	3d	1	0	0	0	0	0	0
10	0	0	3d	1	0	0	0	0	0	0
11	0	0	4de	1	0	0	0	0	0	0
12	0	0	4de	1	0	0	0	0	0	0
13	0	0	4de	1	0	0	0	0	0	0
14	0	0	4de	1	0	0	0	0	0	0

d: desquamation

e: eschar

Interpretation of results:

other: CLP/EU GHS criteria are not met, no classification required according to Regulations (EC) No 1272/2008

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:

01 Sep - 08 Dec 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OTS 798.4500 (Acute Eye Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Hazleton Research Products, Inc., Denver, USA
- Age at study initiation: young adult
- Weight at study initiation: 2317 - 3832 g
- Housing: individual housing in mesh-bottom cages
- Diet: Purina Certified Rabbit Chow #5322, ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.7-20.2
- Humidity (%): 41.2-47.5
- Photoperiod (hrs dark / hrs light): 12/12

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated left eye served as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

Duration of treatment / exposure:

Single treatment without washing

Observation period (in vivo):

72 h

Number of animals or in vitro replicates:

3 per sex

Details on study design:

SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: hand-held penlight / fluorescein

MORTALITY:
The rabbits were observed twice daily (morning and afternoon) for mortality for the duration of the study.

OCULAR OBSERVATIONS:
Both eyes of each rabbit were examined for ocular abnormalities prior to initiation of dosing. The pre-initiation examination included the use of sodium fluorescein and ultraviolet light for detection of corneal abnormalities. The rabbits assigned to study had no pre-existing ocular abnormalities. Both eyes of each rabbit were examined macroscopically for ocular irritation using a handheld penlight in accordance with the method of Draize at approximately one, 24,48 and 72 hours after dosing. In addition, both eyes were further examined at 72 hours with sodium fluorescein and ultraviolet light.

BODY WEIGHTS:
The body weights were obtained and recorded on study day 0 (initiation) and on day 3 (termination).

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritation parameter:

iris score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: not applicable

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0.05

Max. score:

3

Reversibility:

fully reversible within: 72 h

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

of 6 animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

other: not applicable

Irritant / corrosive response data:

After 1 h redness and chemosis (grade 1) were observed in 5/6 and 4/6 animals, respectively. These effects were reversible within 24 h. At 48 h one animal had a redness score of 1, which again was fully reversible upon the next reading time point (72 h). No effects on cornea and iris were observed in any anmal at any time point.

Other effects:

There were no deaths during the study. The rabbits did not vocalize upon instillation of the test material. There were no remarkable changes or differences observed in body weight during the study period.

Table 1: Scores of eye reactions observed in the eye irritation study in rabbits.

Rabbit No.	Time [h]	conjunctivae		iris	cornea
		redness	swelling
1	1	1	1	0	0
	24	0	0	0	0
	48	1	0	0	0
	72	0	0	0	0
	average	0.33	0	0	0
2	1	0	1	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0	0	0	0
3	1	1	1	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0	0	0	0
4	1	1	1	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0	0	0	0
5	1	1	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0	0	0	0
6	1	1	0	0	0
	24	0	0	0	0
	48	0	0	0	0
	72	0	0	0	0
	average	0	0	0	0

Interpretation of results:

other: CLP/EU GHS criteria are not met, no classification required according to Regulations (EC) No 1272/2008

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Assessment of the skin irritating properties of N-[3 -(trimethoxysilyl)propyl]aniline (CAS 3068 -76 -6) was performed based on an acute dermal toxicity study, which was conducted according to OECD 402 and in compliance with GLP (WIL, 1995). Five male and female Crl:CD BR rats were treated with the limit dose of 2000 mg/kg bw by semi-occlusive dermal application for 24 h. The test substance was applied undiluted (1.97 mL/kg) to the back of the animals covering 14-18% of the total body surface. The application sites were wiped with disposable paper towels moistened with deionized water after 24 h. The application sites were examined for erythema, oedema and other dermal findings 30-60 minutes after bandage removal and daily thereafter for thirteen days. The rats were clipped to facilitate dermal observations on study days 7 and 14. Erythema and oedema reactions were scored according to the Draize scoring system, and were thus appropriate for assessment of skin irritating properties as stated in the supplement to the test guideline 404 (A sequential testing strategy for dermal irritation and corrosion).

The rats were observed 1, 3, and 4 h post-dose on day of appplication (day 0) and once or twice daily thereafter for clinical signs and mortality, respectively. Body weights were obtained on the day of treatment and on day 7 and 14. Necropsy of survivors was performed and major organ systems of the cranial, thoracic and abdominal cavities were examined for all animals.

No mortality was observed and no substance-related effects on body weights or necropsy findings were reported. The mean scores over 24, 48, and 72 h were 0 for erythema and edema, calculated over all 10 animals. The test material induced very slight to severe erythema, very slight edema and eschar on one animal, delayed in time. Desquamation was present on 6/10 animals. In one male animal desquamation was observed from day 1 until day 14 without any other findings. In one female animal desquamation was observed from day 3 until day 14. Furthermore erythema occurred at day 5 (grade 1) and increased to grade 2 (day 7+8), 3 (day 9+10), and 4 (day 11 until day 14) in this female animal. Oedema reactions were observed in this animal from day 9 until the end of the study (grade 1). There were no other dermal findings. With the exception of one male and female with severe erythema, slight edema, eschar and/or desquamation, all dermal irritation completely subsided by day 11 or earlier. The effects in the single female animal were judged to be not sufficient for classification. As the effects were observed delayed in time beginning with day 5 the mean value of the animal was calculated over day 5, 6, and 7 for erythema and over day 9, 10, and 11 for oedema, according toRegulation (EC) 1272/2008 (“…if reactions are delayed, from grades on 3 consecutive days after the onset of skin reactions”) resulting in a mean of 1.33 and 1 for erythema and oedema. Thus, neither the mean scores nor the number of animals showing skin reactions until the end of the observation period fulfil the classification criteria according toRegulation (EC) 1272/2008 and according to Directive 67/548/EEC. In addition it has to be taken into account that an exposure period of 24 h may enhance the effects of the test substance as the normal exposure period is 4 h in a standard skin irritation test according to OECD 404.In conclusion, N-[3 -(trimethoxysilyl)propyl]aniline was not irritating to the skin.

An eye irritation test is available with N-[3 -(trimethoxysilyl)propyl]aniline(CAS 3068-76-6), which was performed in New Zealand White rabbits according to OECD 405 and in compliance with GLP (WIL, 1995). Puretest substance (0.1 mL) was instilled into the eye of 3 male and 3 female animals as a single treatment without rinsing of the eye. Both eyes of each rabbit were examined for ocular abnormalities prior to initiation of dosing. The pre-initiation examination included the use of sodium fluorescein and ultraviolet light for detection of corneal abnormalities. The rabbits assigned to study had no pre-existing ocular abnormalities. Both eyes of each rabbit were examined macroscopically for ocular irritation using a handheld penlight in accordance with the method of Draize at approximately 1, 24, 48 and 72 hours after dosing. In addition, both eyes were further examined at 72 hours with sodium fluorescein and ultraviolet light.

No effects on cornea and iris were observed in any animal at any time point. Redness and chemosis (grade 1) were observed in 5/6 and 4/6 animals, respectively, 1 h after instillation of test substance. These effects were reversible within 24 h. At 48 h one animal showed conjunctiva redness (grade 1), which again was fully reversible upon the next reading time point (72 h). The mean values out of all 6 animals over 24, 48, and 72 h were 0, 0, 0, and 0.05 for cornea, iris, chemosis, and conjunctivae redness, respectively. In conclusion, N-[3 -(trimethoxysilyl)propyl]aniline was not irritating to the eyes.

Justification for classification or non-classification

The available data on skin and eye irritation/corrosion of the registered substance do not meet the criteria for classification according to Regulation (EC) 1272/2008 and according to Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.