2-[(3-aminopropyl)methylamino]ethanol

Endpoint summary

Description of key information

Acute toxicity: Oral

In an acute oral toxicity study in female (RccHan™: WIST) rats, following the fixed dose procedure in accordance with the OECD Guideline 420 and EU Method B.1 bis, the LD50 was established to be in the range of 300 - 2000 mg/kg.

Acute toxicity: Inhalation

No reliable acute toxicity study via the inhalation route was available. However, this endpoint is waived as reliable data are available for the oral and dermal exposure routes.

Acute toxicity: Dermal

In an acute dermal toxicity study in male and female New Zealand White rabbits, following the standard acute method according to OECD Guideline 402, the LD50 was established to be in the range of 1000 - 2000 mg/kg (Lemoncelli A, 2007).

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-06-19 to 2017-07-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Version / remarks:

2001

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Version / remarks:

EC NO. 440/2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Specific details on test material used for the study:

Identification: APMMEA
Batch: PFW110370
Purity: 99.10%
Physical state/Appearance: clear colorless liquid
Expiry Date: 31 December 2018
Storage Conditions: room temperature in the dark

Species:

rat

Strain:

Wistar

Remarks:

RccHan™:WIST

Sex:

female

Details on test animals or test system and environmental conditions:

Female Wistar (RccHan™:WIST) strain rats were supplied by Envigo RMS (UK) Limited, Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non pregnant. After an acclimatization period of at least 5 days the animals were selected at random and given a number unique within the study by indelible ink marking on the tail and a number written on a cage card. At the start of the study the animals were 8 to 12 weeks of age. The body weight variation did not exceed ±20% of the mean body weight at the start of treatment.

Animal Care and Husbandry
The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.
The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light and 12 hours darkness.
The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

For the purpose of the 2000 mg/kg dose level, the test item was used as supplied. The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level. For the purpose of the study the 300 mg/kg dose level, the test item was freshly prepared, as required, as a solution in distilled water.
The test item was formulated within 2 hours of being applied to the test system. It is assumed that the formulation was stable for this duration.
No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.

Doses:

300 mg/kg and 2000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

Study Design
Using available information on the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
A single animal was treated as follows:
Dose Level
(mg/kg) Concentration
(mg/mL) Dose Volume
(mL/kg) Number of Rats
Female
300
30 10 1

In the absence of toxicity at a dose level of 300 mg/kg, an additional animal was treated as follows:
Dose Level (mg/kg) Specific Gravity Dose Volume (mL/kg) Number of Rats (Female)
2000 0.964 2.08 1

In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of animals was treated as follows:
Dose Level (mg/kg) Specific Gravity Dose Volume (mL/kg) Number of Rats (female)
2000 0.964 2.08 4

A total of five animals were therefore treated at a dose level of 2000 mg/kg in the study.
Due to mortality at a dose level of 2000 mg/kg, an additional group of animals was treated as follows:
Dose Level(mg/kg) Concentration(mg/mL) Dose Volume (mL/kg) Number of Rats (Female)
300 30 10 4

A total of five animals were therefore treated at a dose level of 300 mg/kg in the study.
All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted body weight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each dose group to confirm the survival of the previously dosed animals.
Clinical observations were made 30 minutes, 1, 2, and 4 hours after dosing and then daily for up to 14 days. Morbidity and mortality checks were made twice daily, early and late during normal working days, and once daily at weekends and public holidays.
Individual body weights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.
At the end of the observation period the surviving animals were killed by cervical dislocation. All animals were subjected to gross necropsy. This consisted of an external examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Data Evaluation
The test item was classified according to Annex 3 of the OECD Guidelines for Testing of Chemicals No. 420 "Acute Oral Toxicity - Fixed Dose Procedure" (adopted 17 December 2001).
Evaluation of data included identification of the number of animals that died during the study (or that were killed for humane reasons), and determination of the nature, severity, onset and duration of the toxic effects. If possible, the signs of evident toxicity were described. Evident toxicity refers to the toxic effects of sufficient severity that administration of the next higher dose level could result in development of severe signs of toxicity and probable mortality. Effects on body weights and abnormalities noted at necropsy were also identified.
Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Dose Level - 2000 mg/kg
Mortality
One animal was killed for humane reasons, 1 day after dosing, due to the occurrence of clinical signs of toxicity that approached the severity limit set forth in the UK Home Office Project License. Two animals were found dead 2 days after dosing.

Dose Level - 300 mg/kg
Based on the results at a dose level of 2000 mg/kg, a dose level of 300 mg/kg body weight was investigated.
Mortality
There were no deaths.

Clinical signs:

Dose Level - 2000 mg/kg
Clinical Observations
No signs of systemic toxicity were noted during the observation period in the initial treated animal.
Signs of systemic toxicity noted in two animals were hunched posture, noisy respiration and pilo erection. Additional signs of systemic toxicity noted in the animal that was humanely killed during the study were ataxia, lethargy, pallor the extremities, emaciation, hypothermia and vocalization.

Dose Level - 300 mg/kg
No signs of systemic toxicity were noted during the observation period.

Body weight:

All animals showed expected gains in body weight over the observation period at both dose levels

Gross pathology:

Dose Level - 2000 mg/kg
Hemorrhagic and ulcerated gastric mucosa was noted at necropsy of the animals that were found dead or humanely killed during the study. No abnormalities were noted at necropsy of the two animals that were killed at the end of the study.

Dose Level - 300 mg/kg
No abnormalities were noted at necropsy

Other findings:

No other findings noted

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 - 2000 mg/kg body weight (Globally Harmonized Classification System - Category 4).

Executive summary:

The study was performed to assess the acute oral toxicity of the test item in the Wistar strain rat.

Methods

Following a sighting test at dose levels of 2000 mg/kg and 300 mg/kg, a further group of four fasted females was given a single oral dose of test item at a dose level of2000mg/kg body weight. Due to mortalities at a dose level of 2000 mg/kg, a further group of four fasted females was given a single oral dose of test item, as a solution in distilled water, at a dose level of 300 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

Results

Mortality. Three animals treated at a dose level of 2000 mg/kg were killed for humane reasons due to the occurrence of clinical signs of toxicity that approached the severity limit set forth in the UK Home Office Project License or found dead, 1 or 2 days after dosing. There were no deaths at a dose level of 300 mg/kg.

Clinical Observations. Signs of systemic toxicity noted in two animals treated at a dose level of 2000 mg/kg were hunched posture, noisy respiration and pilo‑erection. Additional signs of systemic toxicity noted in the animal that was humanely killed during the study were ataxia, lethargy, pallor the extremities, emaciation, hypothermia and vocalization. There were no signs of systemic toxicity noted in the initial animals treated at dose levels of 2000 mg/kg and 300 mg/kg.

Body Weight. Surviving animals showed expected gains in body weight.

Necropsy. Hemorrhagic and ulcerated gastric mucosa was noted at necropsy of the animals that were found dead or humanely killed during the study. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Conclusion

The acute oral median lethal dose (LD₅₀) of the test item in the female Wistar strain rat was estimated to be in the range of 300 ‑ 2000 mg/kg body weight (Globally Harmonized Classification System-Category 4).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2006-10-09 to 2007-01-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: E-015/2005-1
- Expiration date of the lot/batch: no data
- Purity test date: 2005-10-24
- Purity: 99.01 wt% (GC)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Solubility and stability of the test substance in the solvent/vehicle: no data

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan and Robinson Services
- Age at study initiation: 10-18 weeks at start of dosing
- Weight at study initiation: 1.9 to 3.5 kg (± 20% of the mean initial weight) at the outset (Day 1) of the study.
- Housing: Animals were group housed by sex upon receipt and individually housed upon assignment to study in compliance with USDA guidelines. Calvert is a USDA registered and fully AAALAC accredited facility. The room in which the animals were kept was documented in the study records. No other species were kept in the same room.
- Diet (e.g. ad libitum): All animals had access to PMI Certified Hi-Fiber Rabbit Diet #5325 as per Calvert SOP.
- Water (e.g. ad libitum): Tap water was available ad libitum, to each animal via an automatic watering device.
- Acclimation period: a minimum of 5 days prior to dosing.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 to 23°C
- Humidity (%): 23 to 94%
- Photoperiod (hrs dark / hrs light): 12 hours lighU12 hours dark

Type of coverage:

not specified

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area of the trunk
- Type of wrap if used: The test article was applied on an intact skin site for each animal, covered with a gauze patch/dental dam, wrapped with an elastic bandage and secured with non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After exposure the site was unwrapped and wiped with water or mineral oil and gauze to prevent ingestion.
- Time after start of exposure: twenty-four (±0.5) hours.

TEST MATERIAL
- Concentration (if solution): 2 ml/kg

Duration of exposure:

The test article was applied once and remains in contact with the skin site for twenty-four (±0.5) hours.

Doses:

400, 1000 and 2000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/morbidity: once daily;
Clinical observations: Clinical observations were recorded daily through Day 15.
Body weight: Animals were weighed prior to dosing on Day 1 and on Days 8 and 15.
- Necropsy of survivors performed: yes,
All animals were euthanized by CO2 asphyxiation.
The necropsy included examination of the external body surface, all orifices, the cranial, thoracic and abdominal cavities and their contents

Statistics:

The design of this study is such that statistical analysis was not necessary.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 1 000 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

Mortality was not observed in any of the animals at 400 or 1000 mg/kg. One animal was found dead on Day 3 in the 2000 mg/kg group. Based on the clinical signs observed in the remaining animals at 2000 mg/kg they were euthanized on Day 3.

Clinical signs:

Clinical signs observed at 2000 mg/kg were of abnormal gait and stance, decreased activity, decreased body tone and red discolored fur. Clinical signs of abnormal gait and stance were observed during the study at the 400 and 1000 mg/kg dose levels with the addition of poor grooming at 400 mg/kg. No other clinical signs were observed throughout the study. Erythema, edema, necrosis, sloughing and flaking of the skin at the application site were observed during the study at all levels.

Body weight:

No biological effect on body weight was observed in surviving animals at 400 mg/kg. A reduced body weight gain was observed in animals at 1000 mg/kg between Days 1 and 15.

Gross pathology:

No visible lesions were observed in any of the animals dosed with the test article at 400 or 1000 mg/kg at terminal necropsy. Pale stomach, pale stomach lining, pale kidneys and red liquid in the bladders were observed at necropsy of the animals at 2000 mg/kg.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The dermal LD50 of the test substance was estimated to be greater than 1000 mg/kg but less than 2000 mg/kg (2 ml/kg). Based on the CLP criteria, the test substance is classified as acute dermal toxicant category 4.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Additional information

Acute toxicity: Oral

An acute oral toxicity study with the test item according to the fixed dose procedure in female (RccHan™: WIST) rats (OECD guideline 420) was performed (Sanders, 2017) was performed. Following a sighting test at dose levels of 2000 mg/kg and 300 mg/kg, a further group of four fasted females was given a single oral dose of test item at a dose level of 2000 mg/kg body weight. Due to mortalities at a dose level of 2000 mg/kg, a further group of four fasted females was given a single oral dose of test item, as a solution in distilled water, at a dose level of 300 mg/kg body weight. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

Three animals treated at a dose level of 2000 mg/kg were killed for humane reasons due to the occurrence of clinical signs of toxicity that approached the severity limit set forth in the UK Home Office Project License or found dead, 1 or 2 days after dosing. There were no deaths at a dose level of 300 mg/kg.

Signs of systemic toxicity noted in two animals treated at a dose level of 2000 mg/kg were hunched posture, noisy respiration and pilo-erection. Additional signs of systemic toxicity noted in the animal that was humanely killed during the study were ataxia, lethargy, pallor the extremities, emaciation, hypothermia and vocalization. There were no signs of systemic toxicity noted in the initial animals treated at dose levels of 2000 mg/kg and 300mg/kg. Surviving animals showed expected gains in body weight. Hemorrhagic and ulcerated gastric mucosa was noted at necropsy of the animals that were found dead or humanely killed during the study. No abnormalities were noted at necropsy of animals that were killed at the end of the study. The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be in the range of 300 -2000 mg/kg body weight (Globally Harmonized Classification system - Category 4).

 

Acute toxicity: Inhalation

No reliable acute toxicity study via the inhalation route was available. However, this endpoint is waived as reliable data are available for the oral and dermal exposure routes.

Acute toxicity: Dermal

An acute dermal toxicity study with the test substance was performed according to the standard acute method in male and female New Zealand White rabbits (OECD guideline 402; Lemoncelli A, 2007) . One group of ten New Zealand White rabbits (five males and five females) was dermally administered at 2 ml/kg (2000 mg/kg). Based on the results from this group, two more groups were sequentially added and dosed one a single occasion at 400 and 1000 mg/kg. Clinical observations were recorded daily through Day 15. Body weights were recorded on Days 1, 8 and 15. A gross necropsy was performed on Day 15.

Clinical signs observed at 2000 mg/kg were of abnormal gait and stance, decreased activity, decreased body tone and red discolored fur. One male was found dead on Day 3 at 2000 mg/kg and based on the clinical signs the remaining animals were euthanized (moribund sacrificed) on Day 3. No animals died at 400 or 1000 mg/kg during the study. Clinical signs of abnormal gait and stance were observed during the study at the 400 and 1000 mg/kg dose levels with the addition of poor grooming at 400 mg/kg. No other clinical signs were observed throughout the study. Erythema, edema, necrosis, sloughing and flaking of the skin at the application site were observed during the study at all levels. All surviving animals exhibited decreased body weight gains between Days 1 and 15. No visible lesions were observed in any of the animals dosed with the test article at 400 or 1000 mg/kg at terminal necropsy. Pale stomach, pale stomach lining, pale kidneys and red liquid in the bladders were observed at necropsy of the animals at 2000 mg/kg.

The dermal LD50 of the test substance was estimated to be greater than 1000 mg/kg but less than 2000 mg/kg (2ml/kg).

Justification for classification or non-classification

Based on the results, the test substance is classified as acute oral toxicant category 4 and acute dermal toxicant category 4, according to CLP Regulation.