Bis(2-hydroxyethyl)ammonium acetate

Endpoint summary

Description of key information

Studies on DEA acetate were not available, but laboratory animals have been tested with both RA1 and RA2 via the oral and dermal routes. Acute oral LD50 values of 7500 mg/kg bw and over 2000 mg/kg bw were reported for RA2 and RA1, respectively, in rats. Dermal LD50 values in excess of 2000 mg/kg bw were reported for RA1 (in rats) and RA2 (in rabbits). Inhalation exposure is not expected.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

1999-07-21 to 1999-08-13

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 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd.,
Biotechnology & Animal Breeding Division,
CH-4414 Fullinsdorf,
Switzerland.
- Age at study initiation: Males: 8 weeks
Females: 10 weeks
- Fasting period before study: Overnight fasting prior to intubation
- Housing: Groups of three in MAkrolon type -4 cages with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standard Kliba 3433, batch no. 37/99, rat maintenance diet.
- Water (e.g. ad libitum): Community tap water from Itingen
- Acclimatisation: 1 week

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: July 21 -27, 1999 (Females) July 23 - 29, 1999 (males) To: August 11, 1999 (Females) August 13, 1999 (Males)

Route of administration:

oral: gavage

Vehicle:

other: Bi-distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 0.2 g/ml
- Amount of vehicle (if gavage): 10 ml/kg


DOSAGE PREPARATION (if unusual): The test article was placed into a glass beaker on a tared Mettler balance and the vehicle was added. A weight by volume dilution was prepared using a magnetic stirrer as homogenizer. Homogenicity of the test article was in the vehicle was maintained during treatment.

The preparation was made shortly before each dosing.

Doses:

2000 mg/kg body weight

No. of animals per sex per dose:

3 animals per sex. Only one dose (2000 mg/kg bw) was used.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality / viability: four times during test day 1 and once daily during days 2-15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs: Changes in appearance and behaviour were examined four times during day 1 and once daily during days 2-15
- Body weight: Animals were weighed on test day 1 (pre-administration), day 8 and day 15.

Statistics:

No statistical analysis was performed as no deaths occurred.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality

Mortality:

No death occurred during the study.

Clinical signs:

other: No clinical signs were noted during the observation period.

Gross pathology:

No macroscopic findings were observed at necropsy.

Table 1: Individual Findings - Clinical Signs

Sex	Animal No.	Signs	Test Day
			1				2	3	4	5	6	7	8	9	10	11	12	13	14	15
			1h	2h	3h	4h
Female 2000 mg/kg	1	No clinical signs
	2	No clinical signs
	3	No clinical signs
Male 2000 mg/kg	1	No clinical signs
	2	No clinical signs
	3	No clinical signs

Table 2: Body Weights

Body Weight in grams
Sex	Animal No.	Day of Treatment	Day 8	Day 15
Female 2000 mg/kg	1 2 3	175.9 181.4 180.0	204.2 202.9 208.1	217.7 214.9 218.9
Male 2000 mg/kg	1 2 3	206.7 211.7 211.1	264.8 260.1 258.3	289.6 286.4 275.8

Table 3: Macroscopic Findings

Sex	Animal No.	Type of death	Findings
Female 2000 mg/kg	1	Scheduled necropsy	No macroscopic findings
	2	Scheduled necropsy	No macroscopic findings
	3	Scheduled necropsy	No macroscopic findings
Male 2000 mg/kg	1	Scheduled necropsy	No macroscopic findings
	2	Scheduled necropsy	No macroscopic findings
	3	Scheduled necropsy	No macroscopic findings

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

No mortalities or clinical signs of toxicity were seen in male and female rats given a single oral gavage treatment with RA1 at 2000 mg/kg bw, then observed over 14 days. The acute oral LD50 was greater than 2000 mg/kg bw.

Executive summary:

RA1 was evaluated for its acute toxicity potential following oral administration at a dose of 2000 mg/kg bw in male and female Wistar rats.

Animals were administered a single dose of the test substance on a mg/kg bw basis by oral gavage following fasting for approximately 16.5 hours, but with free access to water. Three hours after dosing, the animals were returned to their cages and supplied with feed and water ad libitum.

No mortalities or clinical signs were observed during the study and the body weight of the animals was within the range commonly recorded for this strain and age.

Based on these results, the medial lethal dose (LD50) of RA1 after single oral administration to rats of both sexes, observed over a period of 14 days, was greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

No acute oral data were available on DEA acetate, but two good-quality, reliable, GLP-compliant, OECD-guideline studies on structurally-related compounds RA1 (Rosner, 2000a) and RA2 (Pfeifer, 1996a) meet the REACH tonnage-driven data requirements.

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Quality of whole database:

No relevant data is available on the acute inhalation toxicity of DEA acetate.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

1999-07-29 to 1999-08-19

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 402 (Acute Dermal Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd.,
Biotechnology & Animal Breeding Division,
CH-4414 Füllinsdorf,
Switzerland.
- Age at study initiation: Males: 9 weeks
Females: 12 weeks
- Housing: During Acclimatization: In groups of five in Makrolon type-4 cages with standard softwood bedding.
During treatment and observation: Individually in Makrolon type-3 cages with standard softwood bedding.
- Diet (e.g. ad libitum): Pelleted standarad Kliba 3433, batch no. 37/99, rat maintenance diet.
- Water (e.g. ad libitum): Community tap water from Itingen.
- Acclimation period: 6 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: August 5th 1999 To: August 19th 1999.

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Backs of the animals.
- % coverage: 10%
- Type of wrap if used: Semi-occlusive dressing.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Lukewarm tap water amd dried with disposable paper towels.
- Time after start of exposure: Twenty-four hours after the application.


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.0 ml.
- Constant volume or concentration used: yes

Duration of exposure:

24 hours of exposure with an observation period of 14 days.

Doses:

2000 mg/kg body weight.

No. of animals per sex per dose:

5 males
5 females

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality/viability four times during test day 1 and once daily during days 2 - 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs: Examined for changes in appearance and behaviour four times during day 1 and once daily during days 2 - 15.
- Body weight: On test day 1 (pre-administration), 8 and 15.

Necropsies were performed by experienced prosectors. At the end of the observation period all animals were sacrificed by intreperitoneal injection of NARCOREN at a dose of at least 2.0 ml/kg bw (equivalent to at least 320 mg sodium pentobarbitone/kg body weight). The animals were examined macroscopically and all abnormalities recorded. Thereafter, they were discarded.

Statistics:

No statistical analysis was used as no deaths occurred.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality

Mortality:

No deaths occurred during the study.

Clinical signs:

other: No systemic or local signs of toxicity were observed during the study period.

Gross pathology:

No macroscopic findings were observed at necropsy.

Table 1: Clinical/Local Signs

Sex	Animal No.	Signs	Test Day
			1				2	3	4	5	6	7	8	9	10	11	12	13	14	15
			1h	2h	3h	4h
Female 2000 mg/kg	1	No clinical signs
	2	No clinical signs
	3	No clinical signs
Male 2000 mg/kg	1	No clinical signs
	2	No clinical signs
	3	No clinical signs

Table 2: Body Weights

Body Weights
Sex	Animal No.	Day of Treatment	Day 8	Day 15
Male 2000 mg/kg	1 2 3 4 5	254.6 247.3 262.9 242.8 223.5	296.3 284.3 301.9 268.5 249.2	334.8 322.1 345.7 285.8 270.2
Female 2000 mg/kg	6 7 8 9 10	200.3 202.3 210.3 209.8 205.0	200.4 202.3 208.1 215.5 212.8	205.9 216.1 213.6 226.0 221.5

Table 3: Macroscopic Findings

Sex	Animal No.	Type of death	Findings
Female 2000 mg/kg	1	Scheduled necropsy	No macroscopic findings
	2	Scheduled necropsy	No macroscopic findings
	3	Scheduled necropsy	No macroscopic findings
	4	Scheduled necropsy	No macroscopic findings
	5	Scheduled necropsy	No macroscopic findings
Male 2000 mg/kg	1	Scheduled necropsy	No macroscopic findings
	2	Scheduled necropsy	No macroscopic findings
	3	Scheduled necropsy	No macroscopic findings
	4	Scheduled necropsy	No macroscopic findings
	5	Scheduled necropsy	No macroscopic findings

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The median lethal dose of RA1 after single dermal administration to rats of both sexes, observed over a period of 14 days, could not be estimated as no death occurred. The 24-hr LD50 is greater than 2000 mg/kg bw.

Executive summary:

RA1 was evaluated for its potential to induce acute toxicity following dermal administration at a dose of 2000 mg/kg bw in male and female Wistar rats.

Animals were treated with RA1 at 2000 mg/kg bw by dermal application for 24 hr. The test article was administered undiluted at a volume of 2 ml/kg. The animals were examined for clinical signs and mortality/viability for 14 days after application of the test substance.

No deaths occurred during the study and no clinical signs were observed. In addition, the body weight of the animals was unaffected by treatment and no macroscopic findings were observed at necropsy.

Based on these results, the 24-hr median lethal dose of RA1 after single dermal administration to rats of both sexes is greater than 2000 mg/kg bw.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

No acute dermal data were available on DEA acetate, but two good-quality, reliable, GLP-compliant, OECD-guideline study on structurally-related compounds RA1 (Rosner, 2000b) and RA2 (Pfeifer, 1996b) meet the REACH tonnage-driven data requirements.

Additional information

No acute toxicity data were available on DEA acetate, but reliable guideline studies are available on structurally related compounds.

No mortalities or clinical signs of toxicity were seen in male and female rats given a single oral gavage treatment with RA1 at 2000 mg/kg bw, then observed over 14 days. The acute oral LD50 was greater than 2000 mg/kg bw (Rosner, 2000a). Five of ten treated Sprague-Dawley rats (one male and four females) given RA2 at 7500 mg/kg bw by oral gavage died within 14 days of treatment. The acute oral LD50 was therefore set at 7500 mg/kg bw (Pfeifer, 1996a).

No mortalities or clinical signs of toxicity were seen in rats given a 24-hour dermal application of RA1 at 2000 mg/kg bw and observed for 14 days of treatment. No macroscopic findings were seen at necropsy. The acute dermal LD50 of RA1 was determined to be greater than 2000 mg/kg bw (Rosner, 2000b). A 24-hour dermal application of RA2 at 2000 mg/kg bw caused no clinical signs of toxicity or mortality over the course of the 14-day study in albino rabbits (Pfeifer, 1996b).

According to Regulation (EC) No. 1907/2006, acute toxicity studies need only be provided for the oral route (Annex VII) and, when inhalation exposure of humans likely, the inhalation route (Annex VIII). Based on its low vapour pressure and high water solubility, no inhalation exposure of humans to DEA acetate is expected. As such, the available and reliable oral and dermal data on the read-across substances are considered sufficient for this endpoint.

Justification for selection of acute toxicity – oral endpoint
GLP, OECD guideline study (reliability 2).

Justification for selection of acute toxicity – inhalation endpoint
Data waiver based on the low likelihood of inhalation exposure, and reliable acute oral and dermal data.

Justification for selection of acute toxicity – dermal endpoint
GLP, OECD guideline study (reliability 2).

Justification for classification or non-classification

According to Regulation (EC) No. 1272/2008, classification is applicable when the acute oral or dermal LD50 values are less than or equal to 2000 mg/kg bw. Reliable studies carried out on RA1 and RA2 reported LD50 values in excess of 2000 mg/kg bw (Pfeifer, 1996a,b; Rosner, 2000a,b). On this basis, DEA acetate does not require classification for acute oral or dermal toxicity.

No relevant data is available on the acute inhalation toxicity of DEA acetate. Inhalation exposure is not expected.