Bis(2-ethylhexyl)amine

Endpoint summary

Description of key information

Irritation:
- skin: corrosive (rabbit: BASF 1967, Hoechst Celanese Corporation 1989)
- eye: severe damage (rabbit: BASF 1969, Smyth 1949)

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:

05. Jun 1967 - 13. Jun 1967

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable well documented report which meets basic scientific princples.

Qualifier:

no guideline followed

Principles of method if other than guideline:

BASF-Test:
Four animals were treated for 1, 5, 15 min or 20 hours using occlusive conditions. An application site of 2.5 x 2.5 cm was covered with the liquid test substance. The animals were observed for 8 days and skin changes were recorded daily. The report describes findings after 24 hours and at the end of the observation period (8 days). For a final evaluation, the findings after 48 and 72 hours from the raw data were taken into account.

GLP compliance:

no

Species:

rabbit

Strain:

Vienna White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 3.1 kg (mean)

Type of coverage:

occlusive

Preparation of test site:

not specified

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

no data

Duration of treatment / exposure:

1, 5, 15 min or 20 h

Observation period:

8 days

Number of animals:

4

Details on study design:

TEST SITE
- Area of exposure: 2.5 x 2.5 cm

REMOVAL OF TEST SUBSTANCE
- Washing (if done): concentrated Lutrol and 50% Lutrol
- Time after start of exposure: 1min, 5 min, 15 min and 20 h, respectively

SCORING SYSTEM: The original BASF grading was converted into the numerical grading according to the OECD Draize system.

Irritation parameter:

erythema score

Basis:

mean

Remarks:

animal 1-4

Time point:

24/48/72 h

Score:

2.7

Max. score:

4

Reversibility:

not fully reversible within: 8 days

Irritation parameter:

edema score

Basis:

mean

Remarks:

animal 1-4

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible

Remarks:

8 days

Irritation parameter:

other: necrosis

Basis:

animal: 1-4

Time point:

24/48/72 h

Remarks on result:

other: 15 min treatment: full thickness necrosis (1/4 animals): 7 and 8 days after treatment; necrosis - not full thickness (2/4): 6, 7 and 8 days after treatment and 7 and 8 days after treatment, respectively

Irritant / corrosive response data:

Exposure times of 1, 5 and 15 min caused reddening of the dorsal rabbit skin and severe edema beyond the area of exposure. One week later slight reddening (score 2), rhagades and slight necrosis (1 min and 5 min exposures: not full thickness necrosis), severe necrosis (15 min exposure: full thickness necrosis) and induration of the application site were observed. An application of undiluted bis(2-ethylhexyl)amine, 20 h exposure, led to slight reddening, severe edema and severe necrosis (full thickness necrosis). After 8 days, severe necrosis (full thickness necrosis) and induration of the application site were observed. For details see tables below.

Erythema and edema scores for each animal:

Exposition:	1 min			Comment
Animal	Reading	Erythema	Edema
1	1 min	0	0
2	1 min	2	0
3	1 min	2	0
4	1 min	1	2
1	24 h	3	0	bleedings
2	24 h	3	2
3	24 h	4	3	bleedings
4	24 h	3	3
1	48 h	3	0	bleedings
2	48 h	/	/	not examined
3	48 h	/	/	not examined
4	48 h	3	2	bleedings
1	72 h	2	0
2	72 h	/	/	not examined
3	72 h	/	/	not examined
4	72 h	2	2	bleedings
1	8 days	0	0	necrosis (not full thickness), fissures
2	8 days	3	0
3	8 days	0	0	necrosis (not full thickness), induration
4	8 days	2	0	bluish with yellow regions, induration
mean (animal 1 & 4)*	24 - 48 - 72 h	2.7	1.2
*no mean calculation possible for animals 2 and 3, because scores on the 48 or 72 h readings are missing
Exposition:	5 min			Comment
Animal	Reading	Erythema	Edema
1	5 min	0	0
2	5 min	2	0
3	5 min	2	0
4	5 min	1	2
1	24 h	3	0	bleedings
2	24 h	3	2
3	24 h	3	3
4	24 h	3	3
1	48 h	3	0	bleedings
2	48 h	/	/	not examined
3	48 h	/	/	not examined
4	48 h	2	2	bleedings
1	72 h	2	0
2	72 h	/	/	not examined
3	72 h	/	/	not examined
4	72 h	2	0
1	8 days	0	0	necrosis (not full thickness), fissures
2	8 days	3	2	necrosis (not full thickness)
3	8 days	0	0	anemic necrosis (not full thickness), induration
4	8 days	2	0	bluish with yellow regions, induration
mean (animal 1 & 4)*	24 - 48 - 72 h	2.5	0.8
*no mean calculation possible for animals 2 and 3, because scores of the 48 or 72 h readings are missing
Exposition:	15 min			Comment
Animal	Reading	Erythema	Edema
1	15 min	1	2
2	15 min	3	0
3	15 min	2	0
4	15 min	1	2
1	24 h	3	0	bleedings
2	24 h	3	3
3	24 h	3	3	bleedings
4	24 h	3	3
1	48 h	3	0	bleedings
2	48 h	/	/	not examined
3	48 h	/	/	not examined
4	48 h	2	2	bleedings
1	72 h	3	3	bleedings
2	72 h	/	/	not examined
3	72 h	/	/	not examined
4	72 h	2	2	bleedings
1	8 days	0	0	full thickness necrosis, induration
2	8 days	0	0	necrosis (not full thickness), induration
3	8 days	0	0	anemic necrosis (not full thickness), induration
4	8 days	2	0	bluish with yellow regions, induration
mean (animal 1 & 4)*	24 - 48 - 72 h	2.7	1.7
*no mean calculation possible for animals 2 and 3, because scores of the 48 or 72 h readings are missing
Exposition:	20 h			Comment
Animal	Reading	Erythema	Edema
1	24 h	2	3	bleedings
2	24 h	3	3	induration
3	24 h	2	3	anemic, necrosis (not full thickness)
4	24 h	0	3	full thickness necrosis
1	48 h	2	3	bleedings
2	48 h	/	/	not examined
3	48 h	/	/	not examined
4	48 h	0	3	anemic
1	72 h	2	2	anemic necrosis (not full thickness)
2	72 h	/	/	not examined
3	72 h	/	/	not examined
4	72 h	2	2	full thickness necrosis
1	8 days	2	0	anemic necrosis (not full thickness), bluish with yellow regions, induration
2	8 days	2	0	full thickness necrosis
3	8 days	3	0	full thickness necrosis, induration
4	8 days	0	0	full thickness necrosis, induration, fissures
mean (animal 1 & 4)*	24 - 48 - 72 h	1.3	2.7
*no mean calculation possible for animals 2 and 3, because scores of the 48 or 72 h readings are missing

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

Due to the appearance of full thickness necrosis on day six after treatment (exposure period 15 min) lasting until the end of the observation period of eight days, it can be concluded that the test substance has a corrosive potential.

Executive summary:

Four animals were treated for 1, 5, 15 min or 20 hours using occlusive conditions. An application site of 2.5 x 2.5 cm was covered with the liquid test substance. The animals were observed for 8 days and skin changes were recorded daily. The report describes findings after 24 hours and at the end of the observation period (8 days). For a final evaluation, the findings after 48 and 72 hours from the raw data were taken into account. Due to the appearance of full thickness necrosis on day six after treatment (exposure period 15 min) lasting until the end of the observation period of eight days, it can be concluded that the test substance has a corrosive potential.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable publication which meets standard scientific principles.

Qualifier:

no guideline followed

Principles of method if other than guideline:

Eye injury in rabbits is recorded in a 10-grade ordinal series and is based upon the degree of corneal necrosis that results from instillation of various volumes and concentrations of chemical, as detailed by Carpenter and Smyth. 24 h after instillation the eyes were observed for gross evidence of injury. Grade 1 indicates at most a very small area of necrosis resulting from 0.5 mL of undiluted chemical in the eye. Grade 5 indicates a so-called severe burn from 0.005 mL, and Grade 10 indicates a severe burn from 0.5 mL of a 1% solution in water or propylene glycol.

GLP compliance:

no

Species:

rabbit

Strain:

not specified

Vehicle:

not specified

Controls:

not specified

Duration of treatment / exposure:

24 h

Observation period (in vivo):

24 h

Number of animals or in vitro replicates:

no data

Details on study design:

TOOL USED TO ASSESS SCORE: fluorescein

Irritation parameter:

other: eye injury

Basis:

other: no data

Time point:

24 h

Score:

8

Max. score:

10

Reversibility:

not specified

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The application of the test substance caused serious damage to the exposed eyes.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

05. Jun 1967 - 13. Jun 1967

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable well documented study which meets basic scientific principles.

Qualifier:

no guideline followed

Principles of method if other than guideline:

BASF-Test:
50 µL of the test substance were applied to the conjunctival sac of one eye in 2 animals. The animals were observed after 10 min, 1 and 3 h on the day of treatment and up to 8 days afterwards. Findings were recorded daily. The eyes were not washed out after 24 hours as specified in OECD Guideline 405. The report describes findings after 1 and 24 hours and at the end of the observation period. For final evaluation, the findings after 48 and 72 hours from the raw data were taken into account.

GLP compliance:

no

Species:

rabbit

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 2.85 kg (mean)

Vehicle:

unchanged (no vehicle)

Controls:

other: The adjacent eye served as saline control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 50 µL

Duration of treatment / exposure:

8 days (no washing conducted)

Observation period (in vivo):

8 days

Number of animals or in vitro replicates:

2

Details on study design:

SCORING SYSTEM: The original BASF grading was converted into the numerical grading according to the OECD Draize system.

TOOL USED TO ASSESS SCORE: fluorescein (on day 8 of the observation period)

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

(animal 1-2)

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 8 days

Remarks on result:

other: examination with fluorescein on day 8: a slight opacity was noted at the end of the observation period (8 days) in one animal; see table below for details

Irritation parameter:

conjunctivae score

Basis:

mean

Remarks:

(animal 1-2)

Time point:

24/48/72 h

Score:

1.3

Max. score:

3

Reversibility:

fully reversible within: 7 days

Remarks on result:

other: see table below for details

Irritation parameter:

iris score

Basis:

mean

Remarks:

(animal 1-2)

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

other: no effects observed

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

(animal 1-2)

Time point:

24/48/72 h

Score:

0.48

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

other: see tables below for details

Findings animal 1:

Time	Opacity	Erythema	Chemosis		Iritis
10 min	0	1	0		0
1 h	1	1	2		0
3 h	1	2	2		0
24 h	1	2	1		0
48 h	1	1	0		0
72 h	1	1	0		0
4 d	1	1	0		0
7 d	1	0	0		0
8 d	0	0	0		0

Examination with fluorescein on day 8: corneal opacity score 1

Findings animal 2:

Time	Opacity	Erythema	Chemosis		Iritis
10 min	0	1	0		0
1 h	1	2	1		0
3 h	1	2	2		0
24 h	1	2	2		0
48 h	1	1	0		0
72 h	1	1	0		0
4 d	1	1	0		0
7 d	0	0	0		0
8 d	0	0	0		0

Examination with fluorescein on day 8: no effects

Mean values over 24 h, 48 h and 72 h:

Animal 1: Erythema: 1.3; Opacity: 1; Chemosis: 0.3; Iritis: 0

Animal 2: Erythema: 1.3; Opacity: 1; Chemosis: 0.7; Iritis: 0

Interpretation of results:

irritating

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Additional information

Skin irritation:

In a dermal irritation study (BASF AG 1967; reliability score 2), Vienna White rabbits (4 animals for each exposure period) weredermally exposed to the unchanged test substance for 1, 5, 15 min or 20 hours under occlusive conditions. Animals then were observed for 8 days. The original grading was converted into the numerical grading according to Draize. Exposure times of 1, 5 and 15 min caused reddening of the dorsal rabbit skin and severe edema beyond the area of exposure. One week later slight reddening (score 2), rhagades and slight necrosis (1 min and 5 min exposures: not full thickness necrosis), severe necrosis (15 min exposure: full thickness necrosis) and induration of the application site were observed. An application of undiluted bis(2-ethylhexyl)amine for 20 h led to slight reddening, severe edema and severe necrosis (full thickness necrosis). After 8 days, severe necrosis (full thickness necrosis) and induration of the application site were observed. Hence, these results referred to a corrosive potential of the test substance.

In a second dermal irritation study (Hoechst Celanese Corporation 1989; reliability score 2), the clipped skin of six New Zealand White rabbits was dermally exposed to 0.5 mL of the unchanged test substance for 3 min or 1 hour under occlusive conditions. Animals then were observed for 7 days. Irritation was scored according to the method of Draize. The 3 min exposure led to well defined erythema and minimal to moderate edema (observed at the 60 minute scoring interval). The dermal response progressed to necrosis at 24 hours post exposure in 5/ 6 animals. Blanching was noted on the majority of 3 minute sites at 48 and 72 hours with eschar present by day 7 in 5/6 rabbits. Also the 60 min exposure led to well defined erythema and minimal to moderate edema (observed following patch removal). Additionally, the dermal response progressed to necrosis (after 24 hours), blanching (after 48 and 72 hours) and eschar formation (day 7) in all test animals. Therefore, this study confirmed the corrosive potential of the test substance.

 Additionally, an in vitro test was performed. In the Corrositex test conducted similar to OECD 435 the chemical produced break through time was > 60 min and the test substance was therefore regarded as non-corrosive. Due to the reliable in vivo studies in which the test substance had a corrosive potential this in vitro test was disregarded (Celanese 2002, reliability score 2).

 

Eye irritation:

In a primary eye irritation study (BASF AG 1967; reliability score 2), 0.05 µL of the unchanged test substance was instilled into the conjunctival sac of the eye of each of two rabbits (without washing). Animals then were observed for 8 days. The original grading was converted into the numerical grading according to Draize. The test substance caused corneal opacity (mean of 2 animals (24 - 48 - 72 h): 1). Examinations with fluorescein on day 8 still showed slight opacity in one animal. Conjunctival redness (mean of 2 animals (24 - 48 - 72 h): 1.3) was also observed with fully reversibility within 7 days. Iritis was not seen. Additionally, chemosis was noticed (mean of 2 animals (24 - 48 - 72 h): 0.48) and was reversible within 48 h.

 

The publication of Smyth et al. (1949; reliability score 2) showed, that the application of the test substance for 24 h caused serious damage to the exposed rabbit eyes.

Justification for selection of skin irritation / corrosion endpoint:
One key study available, which is sufficient for assessment.

Justification for selection of eye irritation endpoint:
There are two weight of evidence studies available. They are reliable and the whole database is sufficient for assessment.

Effects on skin irritation/corrosion: corrosive

Effects on eye irritation: corrosive

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

Regarding the whole data on irritation the test substance caused a risk of serious damage to eyes and was corrosive to the skin.

Therefore, classification as skin corrosive (EU-GHS: Cat. 1B) and causing serious eye damage (EU-GHS: Cat. 1) according to Regulation (EC) No.1272/2008 was conducted.