2-[(3-aminopropyl)methylamino]ethanol

Acute Toxicity: oral

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

2001

Deviations:

no

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

2-[(3-aminopropyl)methylamino]ethanol

EC Number:

255-615-9

EC Name:

2-[(3-aminopropyl)methylamino]ethanol

Cas Number:

41999-70-6

Molecular formula:

C6H16N2O

IUPAC Name:

2-[(3-aminopropyl)(methyl)amino]ethan-1-ol

Test material form:

liquid

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

Test animals

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

Administration / exposure

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.

Results and discussion

Effect levels

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

Applicant's summary and conclusion

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).