Bis(2-hydroxyethyl)ammonium acetate

Endpoint summary

Description of key information

No skin or eye irritation data were available on DEA acetate. In reliable studies, read-across substances RA1 and RA2 produced no evidence of skin irritation and only slight transient eye irritation in rabbits. As such, DEA acetate is not expected to be irritating. 

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

1999-07-10 to 2000-04-10

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is the result of a structural analogue substance used as read-across substance. Study is conducted according to Guidelines in a GLP certified laboratory.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

The room temperature should be 20°C±3°C and not 22 ±3°C. This deviation is not thought to have affected the results of this study.

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

The room temperature should be 20°C±3°C and not 22 ±3°C. This deviation is not thought to have affected the results of this study.

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes,
F-01400 Chatillon sur Charlaronne.
- Age at study initiation: Male: 15 weeks
Female: 15 weeks
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls with autoclaved wood and haysticks for chewing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum
- Acclimation period: 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 40 -70%
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light.


IN-LIFE DATES: From: 16-08-1999 To: 19-08-1999

Type of coverage:

semiocclusive

Preparation of test site:

shaved

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml/animal

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3 animals: 1 male, 2 female

Details on study design:

TEST SITE
- Area of exposure: Dorsal area
- Type of wrap if used: Semi-occlusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The area was washed with lukewarm tap water
- Time after start of exposure: 4 hours following application

SCORING SYSTEM:
The skin reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992.

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 1, 24, 48 and 72 hours after removal of application

Score:

0

Max. score:

4

Remarks on result:

other: Reversibility not applicable as no irritation seen

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 1, 24, 48 and 72 hours after removal of application

Score:

0

Max. score:

4

Remarks on result:

other: Reversibility not applicable as no irritation seen

Irritant / corrosive response data:

The test article did not elicit any skin reactions at the appplication site at any of the observation times. The mean values from 24 to 72 hours were therefore 0/4 for erythema and 0/4 for oedema.

No irreversible alterations of the treated skin were observed nor were corrosive effects evident on the skin.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the test and observation period, and no mortality occurred.
No staining by the test article of the treated skin was observed.
The body weights of all rabbits were considered to be with in the normal range of variability.

Table 1: Body Weights

Body Weights in grams
Animal No.	Sex	First Day of Acclimatization	Day of Treatment	Last Day of Observation
10	Male	2585	2752	2782
11	Female	2503	2681	2679
12	Female	2579	2612	2668

Table 2: Skin Irritation Scores - Individual Values

Animal Number	Sex	Evaluation Interval	Erythema	Oedema
10 11 12	Male Female Female	1 hour	0 0 0	0 0 0
10 11 12	Male Female Female	24 hours	0 0 0	0 0 0
10 11 12	Male Female Female	48 hours	0 0 0	0 0 0
10 11 12	Male Female Female	72 hours	0 0 0	0 0 0

Table 3: Skin Irritation Scores - Mean values of 24, 48 and 72 hours

Animal Number	Sex	Erythema          N	Oedema             N
10 11 12	Male Female Female	0.00                   3 0.00                   3 0.00                   3	0.00                   3 0.00                   3 0.00                   3

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the classification criteria, RA1 is considered to be "not irritating" to rabbit skin following a 4-hr application.

Executive summary:

RA1 was tested for its dermal irritancy potential. 0.5 ml of test substance was applied to the skin of 3 White New Zealand rabbits for 4 hours using a semi-occlusive dressing. Following removal of the dressing and washing off the test substance, the animals were observed for 72 hours for signs of skin irritation, clinical signs, and changes in body weight.

The test article did not elicit any skin reactions at the appplication site at any of the observation times. The mean values at 24, 28 and 72 hours were 0/4 for erythema and 0/4 for oedema. The test substance did not induce any clinical signs in the treatment animals and all animals gained body weight during the course of the study.

Based on these results, RA1 is not considered to be a skin irritant.

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:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Study period:

1999-08-17 to 1999-08-30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is the result of a structural analogue substance used as read-across substance. Study is conducted according to Guidelines in a GLP certified laboratory.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage Scientifique des Dombes,
F-01400 Chatillon sur Charlaronne,
France.
- Age at study initiation: Males: 15 weeks
Females: 14 weeks
- Weight at study initiation: Male 1 = 2737 g, Female 1 = 2917 g, Female 2 = 2974 g
- Housing: Individually in stainless steel cages equipped with feed hoppers, drinking water bowls, with wood and haysticks for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum
- Water (e.g. ad libitum): Community tap water ad libitum
- Acclimation period: Five days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 50-80%
- Air changes (per hr): 10 -15
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light.


IN-LIFE DATES: From: 17th August 1999 To: 30th August 1999

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml/animal

Duration of treatment / exposure:

The test substance was applied once into one eye of each rabbit. The treated eyes were not rinsed after application. Scoring of irritation effects was performed up to 7 days after test substance application.

Observation period (in vivo):

There was a 7 day observation period after the application of the substance.

Number of animals or in vitro replicates:

Three animals: One male and two females.

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eye was not rinsed following application


SCORING SYSTEM:
The ocular reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992 at approximately 1, 24, 48 and 72 hours, as well as 7 days after administration.

TOOL USED TO ASSESS SCORE: Varta Cliptrix diagnostic-lamp.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1, 24, 48 and 72 hour, and 7 days

Score:

0

Max. score:

4

Remarks on result:

other: Reversibility not applicable as no irritation seen

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 hour to 7 days

Score:

0

Max. score:

2

Remarks on result:

other: Reversibility not applicable as no irritation seen

Irritation parameter:

conjunctivae score

Remarks:

chemosis

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 24 hours

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 1 hour

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 7 days

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 24, 48 and 72 hours

Score:

0.33

Max. score:

13

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Irritation:
Application of the test article to healthy rabbit conjunctiva resulted in a primary irritation score of 0.33 (out of 13). Slight reddening and slight swelling of the conjunctiva was noted in all animals one hour after treatment. Slight reddening of the conjunctiva persisted 72 hours after the administration in one animal and disappeared on the 7-day reading. The sclera of one animal was slightly reddened at the 1-hour examination.

Corrosion:
No corrosion of the cornea was observed at any of the reading times.

Other effects:

Viability/Mortality and Clinical Signs:
No clinical signs of systemic toxicity were observed in the animlas during the test and observation period, and no mortality occurred.

Coloration:
No staining by the test article was observed.

Body Weights:
The body weight of all rabbits were considered to be within the normal range of variability for the species.

Table 1: Body Weights:

Body Weights in grams
Animal No.	Sex	First Day of Acclimatization	Day of Treatment	Last Day of Observation
1	Male	2737	2903	3118
2	Female	2917	3107	3291
3	Female	2974	2840	3095

Table 2: Eye Irritation Scores - Individual Values:

Animal Number	Sex	Evaluation Interval	Corneal Opacity	Iris	Conjunctivae		Cumulative
					Redness	Chemosis	Score	Mean
1 2 3	Male Female Female	1 hour	0 0 0	0 0 0	1 1 1	1 1 1	2.00 2.00 2.00	2.00
1 2 3	Male Female Female	24 hours	0 0 0	0 0 0	0 1 0	0 0 0	0.00 1.00 0.00	0.33
1 2 3	Male Female Female	48 hours	0 0 0	0 0 0	0 1 0	0 0 0	0.00 1.00 0.00	0.33
1 2 3	Male Female Female	72 hours	0 0 0	0 0 0	0 1 0	0 0 0	0.00 1.00 0.00	0.33
1 2 3	Male Female Female	7 days	0 0 0	0 0 0	0 0 0	0 0 0	0.00 0.00 0.00	0.00

Table 3: Eye Irritation Scores - Mean Values after 24, 48 and 72 hours:

Animal Number	Sex	Corneal    Opacity	Iris	Conjunctivae		Primary Eye Irritation Score
				Redness	Chemosis
1 2 3	Male Female Female	0.00          0.00          0.00	0.00    0.00 0.00	0.00       1.00      0.00	0.00               0.00               0.00	0.33

Table 4: Eye Irritation Scores - Mean Values over Time:

Evaluation Interval	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
1 hour	0.00	0.00	1.00	1.00
24 hours	0.00	0.00	0.33	0.00
48 hours	0.00	0.00	0.33	0.00
72 hours	0.00	0.00	0.33	0.00
7 days	0.00	0.00	0.00	0.00

Table 5: Eye Irritation Scores - Assessment according to EC Guidelines:

Evaluated Interval	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
24 hours 48 hours 72 hours	Not Irritating	Not Irritating	Not Irritating	Not Irritating

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In a GLP guideline study, RA1 was considered to be not irritating to the rabbit eye.

Executive summary:

RA1 was tested for its eye irritation potential. 0.1 ml of the undiluted test substance was placed in the conjunctival sac of the left eye of each animal. The lids were gently held together for about 1 second to prevent loss of the test article. The right eye remained untreated and served as the reference control.

The animals were observed for 72 hours for signs of eye irritation, clinical signs, mortality and changes in body weight.

Application of the test article to healthy rabbit conjunctiva resulted in a primary irritation score of 0.33 (out of 13). Slight reddening and slight swelling of the conjunctiva was noted in all animals one hour after treatment. Slight reddening of the conjunctiva persisted 72 hours after the administration in one animal and disappeared on the 7-day reading. The sclera of one animal was slightly reddened at the 1-hour examination.

No corrosion of the cornea was observed at any of the reading times. The test substance had no effect on viability, clinical signs or body weight.

Based on these results, RA1 is considered not to be an eye irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

No data are available on the dermal or eye irritancy of DEA acetate, but studies are available on structurally-related read-across compounds.

 

Skin irritation was not seen in New Zealand White rabbits within 72 hr of a 4-hr semi-occlusive application of RA1. Mean scores of 0/4 were seen for both erythema and oedema from 24 to 72 hr (Arcelin, 2000a). RA2 similarly gave no evidence of erythema or oedema when applied to the skin of the same strain of rabbits for 4 hours, with animals observed for 72 hours (Prezioso, 1996a).

 

Eye irritancy was investigated when 0.1 ml of undiluted RA1 was applied to the left eye of New Zealand White rabbits. Mild irritative effects, resolving within 7 days, were seen in all animals. An average primary irritation score of 0.33/13 was observed between 24 and 72 hr after treatment. On this basis, RA1 was not considered to be corrosive or an eye irritant (Arcelin, 1999). Treatment with 0.1 ml RA2 similarly caused only mild conjunctival effects in the eyes of New Zealand White rabbits 1 hour after administration (with a score of 6/20). These effects were fully reversible within 24 hours, and RA2 was not considered to be irritating or corrosive to the eyes of rabbits (Prezioso, 1996b).

Justification for selection of skin irritation / corrosion endpoint:
GLP, OECD guideline study (reliability 2).

Justification for selection of eye irritation endpoint:
GLP, OECD guideline study (reliability 2).

Justification for classification or non-classification

According to Regulation (EC) No. 1272/2008, classification is applicable when at least 2 of 3 tested animals have a mean skin irritation score of between 2.3 and 4.0 between 24 and 72 hr after treatment. Read-across compounds were not significantly irritating to the skin (Arcelin, 2000a; Prezioso, 1996a) and were not classified as irritating to the eyes of rabbits (Arcelin, 1999; Prezioso, 1996b). On this basis, DEA acetate does not require classification as a skin, eye or respiratory irritant (or as corrosive).