N,N-bis(3-aminopropyl)methylamine

Endpoint summary

Description of key information

The test substance bis-(3-aminopropyl-)methylamine showed indications of toxicity in acute toxicity studies in rats (oral and inhalation routes) and in rabbits (dermal route). Toxicity was reflected in the following LD50 values obtained: LD50 oral - 691 mg/kg bw, LD50 dermal - 127 mg/kg bw and LC50 inhalation 0.07 mg/m³ and in the clinical signs observed (systemic and local/ corrosive effects). 

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:

1979-06-20 until 1979-07-24

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP, no guideline was followed/ mentioned (but similar to OECD 401 guideline)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: By males an average weight of 262.5 g, by females an average weight of 207.5 g
- Fasting period before study: 15 h - 20 h before administration
- Diet: Herilan MRH-Haltung ; H. Eggersmann KG

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 10.0, 6.81, 4.64, 3.16 % (w/v)
- Amount of vehicle (if gavage): 10 mL/kg

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

Doses:

1000, 681, 464, 316 mg/kg

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: For mortality -1hour, 1, 2, 7, 14 days, for weighing 2 - 4 days, 7, 13 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology

Statistics:

Chi - 2 test

Sex:

male/female

Dose descriptor:

LD50

Effect level:

691 mg/kg bw

Based on:

test mat.

Mortality:

At the dose group of 316 mg/kg bw 0 % mortality after 14 days of observation
At the dose group of 464 mg/kg bw 10 % mortality after 14 days of observation
At the dose group of 681 mg/kg bw 60.0 % mortality after 14 days of observation
At the dose group of 1000 mg/kg bw 80 % mortalityafter 14 days of observation.

Clinical signs:

other: At dose groups 464, 691 and 1000 mg/kg bw animals were in poor general state and the following signs and symptoms were observed: dyspnea, apathy, staggering, spastic gait, piloerection, erythema and salivation.

Gross pathology:

Necropsy findings by animals that died:
Heart: acute dilatation ; acute passive hyperemia
Liver: peripheral clay-colored acinar pattern
Intestines : atonic and diarrheal contents
Kidneys : pale in several animals

Necropsy findings by sacrificed animals:
No abnormalities were detected.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

691 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1978

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP, no guideline was indicated (but similar to OECD guideline 403)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: WIGA, Sulzfeld Germany
- Weight at study initiation: 185 + 15 g
- Diet: Food from Herilan MRH from the firma H . EGGERSMANN KG, Rinteln/Weser
- Water: Tap water ad libitum

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose/head only

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
A dynamic inhalation head - nose exposre system. An infusion pump (UNITA i, B. BRAUN, Melsungen) that constantly assist in supplying aerosols.

TEST ATMOSPHERE
- Brief description of analytical method used: Gaschromatographie
- Samples taken from breathing zone: yes

CLASS METHOD
- Rationale for the selection of the starting concentration: Based of an "inhalation risk test" the doses were selected aimed to include a no effect level (0 mortality).

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Nominal: 0.3, 0.104, 0.1, 0.06 mg/L
Measured (analyt.): 0.12, 0.085, 0.05, 0.035 mg/L

No. of animals per sex per dose:

10 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical signs and mortality were observed every day. Weighing was performed on day 7 and at the end of the observation period
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology.

Statistics:

Statistics was performed according to D .J . Finney; Probitanalysis 1971, p. 1- 150.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

0.07 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

Mortality was observed at dose groups (nominal) 0.1 mg/L (mortality of male animals 7/20 and 3/20 mortality of female animals, animals died with in a period of 14 days), 0.104 mg/L (mortality of male animals 2/10 and 5/10 mortality of female animlas, animals died within a period of 14 days) , 0.3 mg/L (mortality of male animals 10/10 and 10/10 mortality of female animals, animals died within a period of 8 days).

Clinical signs:

other: Symptoms of exposure observed were clear to reddish eye and nasal discharge, changed breathing pattern, respiratory sounds, reduced movements and squatting posture, staggering and high stepping gait, and ruffled fur.

Body weight:

Body weight development of the female animals exposed to the low concentration was not influenced. Body weight development in the surviving male animals of the other groups was depressed in the first post exposure week in a concentration dependent manner, but reeovered in the second week.

Gross pathology:

During necrapsy the animals that died showed dilation of aurieles, acute general congestion, hyperemia, edema and emphysema of the lungs as well as lobular pattern in the Iiver. Partly carnified lungs were noted in some surviving animals examined at the end of the study.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

70 mg/m³ air

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:

1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP, no guideline followed but similar to the OECD 402 guideline with deviations.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

yes

Remarks:

, less than 5 animals per dose (4 animals max. were used per dose). In the oldest study 2 dose levels were examined instead of 3 dose levels.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rabbit

Strain:

New Zealand White

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 3 - 5 months
- Weight at study initiation: average 2.5 kg
- Diet: ad libitum, Rockland standard rabbit diet
- Water: ad libitum

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Duration of exposure:

24 hours

Doses:

In the oldest study (1957), 2 doses were used, 0.10 mL/kg bw and 0.20 mL/kg bw of the undiluted test substance.
In the latest study (1974) 3 doses were used, 0.20 mL/kg bw, 0.40 mL/kg bw and 0.80 mL/kg bw of the undiluted test substance.

No. of animals per sex per dose:

In the oldest study (1957) 4 male animals per dose were examined.
in the latest study (1974) two dose groups consisted of 4 male animals per dose and one group with 2 animals (the group with the highest concentration examined)

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, indications on the skin, gross pathology

Statistics:

Thompson´s method of calculating LD50 was used.

Sex:

male

Dose descriptor:

LD50

Effect level:

0.14 mL/kg bw

Based on:

test mat.

Remarks on result:

other: equivalent to LD50=127 mg/kg bw. The result was obtained in the oldest study (1957)

Sex:

male

Dose descriptor:

LD50

Effect level:

0.252 mL/kg bw

Based on:

test mat.

Remarks on result:

other: Equivalent to LD50=228 mg/kg bw. The result was obtained in the latest study (1974).

Mortality:

Oldest study (1957):
In the dose group 0.10 mL/kg bw no mortality was observed.
In the 0.20 mL/kg bw dose group mortality of 4/4 within 1 -3 days (100 % mortality)
Latest study (1974):
In the 0.20 mL/kg bw dose group mortality of 1/4
in the 0.40 mL/kg bw dose group mortality of 4/4 (100 % mortality)
In the 0.80 mL/kg bw dose group mortality of 2/2 (100 % mortality)

Clinical signs:

other: On the skin applied with the test substance marked skin necrosis was observed. Symptoms as unsteady gait and lethargic were indicated in the later study (1974).

Gross pathology:

Oldest study (1957): lung hemorrhage, pale or yellow coloration of livers, pale yellow kidneys with potential hemorrhage.
Latest study (1974): by animals that died during the exposure/ observation period livers and kidneys were pale and mottled , lungs and spleens congested. No gross pathological indications were observed by survivors.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

127 mg/kg bw

Additional information

Acute toxicity: oral

In the most robust acute oral study available (BASF, 77/438), the test substance bis-(3-aminopropyl-)methylamine was examined in an acute oral toxicity study performed similarly to the OECD 401 guideline with 5 (Sprague - Dawley) rats per sex per dose. The rats were administered by gavage with test substance doses of 1000, 681, 464 and 316 mg/ kg bw and observed for 14 days for mortality clinical signs and gross pathology indications. Mortality occurred at dose groups, 464 mg/kg (10 %), 681 mg/kg bw (60 %) and 1000 mg/kg bw (80 %) and the following clinical signs were observed: poor general state, dyspnea, apathy, staggering, spastic gait, piloerection, erythema and salivation. By animals that died during the observation time the following indications were observed in autopsy: acute dilatation and acute passive hyperemia in the heart, peripheral clay - colored acinar pattern in the liver, atonic and diarrheal contents in the intestines and pale kidneys in several animals. Based on the mortality results a LD50 value of 691 mg/kg bw was determined.

Other studies, less detailed ones which were defined as supporting studies showed mostly a higher LD50 value than the one obtained in the key study (worst – case). In an acute oral study (Myers, 1996 and NTS/OTS, 1992) with the test substance bis-(3-aminopropyl-)methylamine, performed similarly to the OECD 423 guideline as part of a project that aimed to compare among other tox parameters acute toxicity of various classes of amines. 5 male Wistar rats per dose were examined by gavage. The test substance was given intially undiluted. Dosages were varied by a constant factor (of two) until sufficient mortality was obtained for an LD50 determintaion. The exact dosage was not indicated. Application of the test item was followed by 14 days observation period. An LD50 value of 1.54 mL/kg bw equivalent to 1356 mg/kg bw was determined. After dosing the animals were slaggish and in autopsy the following indications were observed: slight lung congestion, gastrointestinal tract congestion, hemorrhage and burning, congestion of adrenals and mottling of livers.

In another supporting study (BASF, 1978) the test substance Bis-(3 -aminopropyl-)methylamine in concentrations between 0.5 - 20 % (in aqua dest.) was examined. A LD50 value > 200 and < 1000 mg/kg bw was determined.

In the following supporting study (BASF, XIII44) the test substance Bis-(3 -aminopropyl-)methylamine was examined. Rats were applied with 20 % solution of the test substance (p.o). After 7 days the following LD50 values were determined:

LD50 = 995 mg/kg bw (1.1 mL/kg bw), obained in rats with 20 % pH 11 test solution and LD50 = 904 mg/kg bw (1.0 mL/kg bw), obtained in rats with 20 % pH 7 test solution.

Symptoms observed: staggering, slowed breathing rythem. In autopsy: Following application of the basic test substance solution, corrosion of the peritoneaum was observed.

Acute toxicity: inhalation

In the most substantial acue inhalation study (BASF, 1978) with the test substance Bis-(3-aminopropyl-)-methylamine (key study), performed similarly to OECD 403 guideline, 10 male and 10 female rats (Sprague-Dawley) were exposed to test substance aerosols in the following doses: 0.035, 0.05,0.085 and 0.12 mg/L (measured, analytically determined). Mortality incidences were observed in the following doses: 0.12 mg/L (mortality of male animals 7/20 and 3/20 mortality of female animals), 0.085 mg/L (mortality of male animals 2/10 and 5/10 mortality of female animlas), 0.05 mg/L (mortality of male animals 10/10 and 10/10 mortality of female animals). The LC50 for male and female animals was evaluated to be 0.07 mg/L. Body weight development of the female animals exposed to the low concentration was not influenced. Body weight development in the surviving animals of the other groups was decreased in the first post exposure week in a concentration dependent manner, but recovered in the second week. Symptoms of exposure observed were clear to reddish eye and nasal discharge, changes in the breathing pattern and heaviliy breathing, reduced movements and squatting posture, staggering and high stepping gait, and ruffled fur. During necropsy the animals that died showed dilation of aurieles, hyperaemia, edema and emphysema of the lungs as well as lobular pattern in the Iiver. Partly carnified lungs were noted in some surviving animals examined at the end of the study.

In supporting studies no LC50 could be determined due to technical problems.

In the following supporting study (NTIS/OTS, 1992) three studies were reported with the test substance N, N bis(3 -aminopropyl)methylamine. In the first study reported (1957), 6 rats were exposed to saturated vapour, dynamic conditions at room temperature. After 6 hours the sample became more viscous and milky in appearance probably indicating a reaction with carbon dioxide or with water in the air. This reaction may have reduced the toxic effects of the material therefore the concentration probably did not exceed 5 to 10 ppm) and since the vapor pressure is low, around 0.02 mm Hg at 20°C, it suggests that the test substance is not hazardous, by inhalation, under normal handling condition. No mortality was observed. In the 1974 study, 6 rats were exposed to saturated vapour, static conditions, 8 hours exposure. No mortality was observed. In the other 1957 study, 6 rats were exposed to an exceeded concentration concentration of 2 mg/L (330ppm) of the test substance. Since this concentration was well above saturation, it is likely that the animals were exposed to an aerosol rather than a vapour. Mortality of 100 % was observed after 1 hour of exposure (LC100 = 2 mg/L) and lung hemorrhage was observed in necropsy. No LC50 and thus, no classification could be determined based on the results reported.

In the supporting study (Myers, 1996) an acute inhalation study with the test substance was performed as part of a project that aimed to compare among other tox parameters acute toxicity of various classes of amines. Six female rats were exposed for 8 hours to the test substance by a static saturated vapour exposure. 50 to 100 L of the test material was placed in a 120 L sealed chamber and the atmosphere allowed to equilibrate overnight (approximately 16 h) . The animals were then rapidly introduced into the chamber by means of a gasketed drawer-like system designed to minimize vapor loss. The concentration was not indicated and the endpoint value measured was TL50 (median lethal time). As no mortality was observed after 14 days of observations, no more concentrations were examined (therefore indicated above as limit test) and a TL50 value of > 8 hours was determined. Additionally, until the end of the observation period no clincal signs and and no unusual findings were observed.

Acute toxicity: dermal

The key study (the most appropriate documentation of results, (NTIS/OTS, 1992) included two reported acute dermal studies performed with the test substance Bis-(3-aminopropyl-)methylamine. In both studies (similarly to OECD 402 guideline with some deviations) male albino rabbits were applied with the test substance for 24 hours and observed for 14 days. In the oldest study (1957), two dose groups of 4 rabbits each were administered with 0.1 mL/kg bw and 0.2 mL/kg bw of the undiluted test substance while in the latest study (1974) 2 dose groups of 4 rabbits each were adminstered with the undiluted test substance in concentrations of 0.20 mL/kg bw and 0.40 mL/kg bw and one dose group of 2 rabbits were administered with 0.80 mL/kg bw. Results of the oldest study (1957) showed no mortality (0/4 mortality) in the 0.10 mL/kg bw dose group but 100 % mortality (4/4 mortality) in the 0.2 mL/kg bw dose group within 1- 3 days. In the autopsy lung and possible kidney hemorrhage were observed and pale and yellow liver and kidneys. Additionally the skin applied with the test substance showed marked necrosis. Based on the results of this study an LD50 value of 0.14 mL/kg bw (equivalent to 127 mg/kg bw) was determined. In the latest study (1974), at dose level of 0.20 mL/kg bw 1 animal of 4 died (1/4 mortality, in contrast to the result of the older study) and 100 % mortality (4/4) occured in dose groups 0.40 and 0.80 mL/kg bw. Necrosis of the applied skin was observed in all dose levels and by animals that died during the exposure/ observation period. Pale mottled liver and kidneys and congested lungs and spleens were observed in autopsy. Additionally the skin applied with the test substance showed marked necrosis and other symptoms as unsteady gait and lethargic were indicated. Based on the results of this study a LD50 value of 0.252 mL/kg bw (equivalent to 228 mg/kg bw) was determined. The difference in the LD50 values between the two studies is probably originated from the inconsistence in the mortality observed in dose level 0.2 mL/kg bw (1957 study: , mortality of 4/4, 1974 study: mortality of 1/4)

As both studies seemed to be of the same quality, it is then expected to select the the worst - case result, thus, LD50=0.14 mL/kg bw (127 mg/kg bw), obtained in the oldest study (1957).

Results of the supporting studies were in line with the results of the key study.

In an acute dermal study (supporting study, BASF 77/438, 1978) performed with the test substance Bis-(3-aminopropyl)-methylamine, rabbits were exposed to the undiluted test substance and observed for 4 days. The LD50 value determined was 200 mg/kg bw. The animals behaved apathicly and the following indications were observed in autopsy of animals that died during the application or observation period: heart - myodegeneration, and dilatation, liver - fat liver, degeneration. No gross pathological indications were observed by survivors. In the supporting study (BASF, XIII44) two experiments were described. In the firstly described experiment the back and the ears of the 4 rabbits were applied with the undiluted test substance, exposre time as well as the amount applied were not indicated. In the secondly described experiment 210 cm² of the back of 3 rabbits were applied with 1 mL/kg bw of the undiluted test substance (exposure time not indicated) and 50 cm² were applied with 0.2 mL/kg bw of the undiluted test substance for 4 hours. Strong skin necrosis and strong oedema and redeness were observed in all animals. In the firstly described experiment and in the second described experiment (with 1 mL/kg of the undiluted test substance) all animals died within 3 days. The following clinical and pathological signs were observed: Bloody urine with high protein content, disturbed blood picture, urea in serum, damage observed in the pathological examination to the liver kidney and bladder.

Justification for classification or non-classification

Based on the results obtained the test substance is classified as Cat. 4 H302 for acute oral toxicity, Cat. 2, H330 for acute inhalation toxicity and Cat. 2 H310 for acute dermal toxicity according to Regulation (EC) No 1272/2008 (CLP).