3,3'-[butane-1,4-diylbis(oxy)]bispropanamine

Endpoint summary

Description of key information

LD50 oral = 3450 mg/kg (BASF, 1972; OECD401)
LC50 inhalation (4h, aerosol) = 1.5 mg/L (BASF1980; OECD403)
LD50 dermal > 1000 (male ) and > 2000 mg/kg (female) (BASF 2001; OECD402)

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Principles of method if other than guideline:

In principle, the methods described in the OECD Guideline 401 were used. Young adult laboratory rats were purchased from breeder. Usually the source and strain of animals were not documented. Several groups of 5 rats per sex and dose were treated simultaneously by gavage with preparations of the test substance in suitable vehicle. The concentrations of these preparations were used to achieve comparable volumes per kg body weight. Group-wise documentation of clinical signs was performed over the 7 day study period. Body weight was determined before the start of the study only, as it was needed for determination of dose.

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Species:

rat

Strain:

not specified

Sex:

male/female

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

2% aqueous solution

Doses:

7200, 5600, 4500, 3600, 2800 and 2250 mg/kg bw

No. of animals per sex per dose:

5 per sex and dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 7 days;
- The animals were observed for mortality and clinical signs of toxicity;
- Frequency of observations: Several times on the application day, thereafter once each working day;
- Body weights were only recorded at the beginning of the study;
- Necropsy of survivors and animals which died performed: yes

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

ca. 3 450 mg/kg bw

Based on:

test mat.

Remarks on result:

other: calculated from original value ca. 3600 µl/kg bw using density of 0.96 g/cm³

Mortality:

see table below

Clinical signs:

other: No symptoms monitored in rats

Gross pathology:

Dead animal: dilatation of the heart, stomach filled with liquid and dilated; extensive hemorrhagic erosions; sanguineous, diarrhetic stomach content
Surviving animals: no findings

Mortality data:

Dose [cmm/kg]	Mortality
	Male	Female
7200	5/5	5/5
5600	5/5	5/5
4500	4/5	4/5
3600	2/5	2/5
2800	0/5	0/5
2250	1/5	1/5

Interpretation of results:

Category 5 based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

3 450 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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

GLP compliance:

no

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

- Analytical purity: approximately 99%

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: MUS RATTUS, Brunnthal
- Weight at study initiation: 185 + 15 g
- Diet: Herilan MRH
- Water: ad libitum

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose/head only

Vehicle:

air

Details on inhalation exposure:

The test substance was applied in a constant quantity with the help of an infusion pump to a 2-step blast pipe. Synthetic air (2.6 bar) was used to generate an aerosol, which was infused into the system.

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

22.9, 10.01, 2.19 and 0.41 [mg/L] (nominal concentration)
3.72, 1.76, 0.81, 0.17 [mg/L] (analytic concentration)

No. of animals per sex per dose:

10

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: weekly
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Statistics:

according to D.J. Finney (D..J*. Finney; Probitanalysis 1971, Seite 1 - 150). Published by the Syndics of the Cambridge University Press, Bentley House, 200 Euston Road, London N.W. 1

Sex:

male/female

Dose descriptor:

LC50

Effect level:

1.5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Mortality:

see table below; the animals died within 1-5 days after test item application

Clinical signs:

other: 3.72, 1.76 [mg/L] (analytic concentration): reddish head, dyspnea, scrubby, shaggy fur, tremor 0.81 [mg/L] (analytic concentration): accelerated respiration 0.17 [mg/L] (analytic concentration): nothing abnormal detected Surviving animals did either not

Body weight:

The only male animal surviving exposure to 3.75 mg/l showed a significantly reduced body weight; furthermore, males at 1.76 mg/l showed some reduction on body weight, whereas all other animals exhibited body weights similar to control animals 14 days subsequent to exposure. (see table below)

Gross pathology:

heart: acute dilatation
lung: strong hyperemia
kidney: some animals showed tubule necrosis

Analytic concentration	Nominal concentration	Mortality males	Mortality females
3,72	22,9	9/10	10/10
1,76	10,01	6/10	6/10
0,81	2,19	0/10	3/10
0,17	0,41	0/10	0/10

Analytic concentration	Mean body weight d0		Mean body weight d7		Mean body weight d14
	males	females	males	females	males	females
3,72	186 (10)	183 (10)	124 (1)	all dead	151 (1)	all dead
1,76	179 (10)	180 (10)	211 (4)	181 (4)	238 (4)	214 (4)
0,81	179 (10)	179 (10)	213 (10)	186 (7)	266 (10)	207 (7)
0,17	182 (10)	180 (10)	225 (10)	201 (10)	269 (10)	214 (10)
Control	182 (10)	182 (10)	218 (10)	199 (10)	264 (10)	213 (10)

The number in brackets indicates the number of animals alive at the respective time point.

Interpretation of results:

Category 4 based on GHS criteria

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LC50

Value:

1 500 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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

GLP compliance:

yes

Test type:

standard acute method

Limit test:

no

Specific details on test material used for the study:

- Purity: 99.7 %

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Boehringer Ingelheim Pharma KG
- Age at study initiation: young adult animals
- Weight at study initiation: 200-300 g +/- 20% of the mean weight
- Housing: single housing
- Diet (e.g. ad libitum): Kliba-Labordiaet ad libitumn
- Water (e.g. ad libitum): Tap water ad libitumn per day.
- Acclimation period: at least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%

- Photoperiod (hrs dark / hrs light): 12/12

Type of coverage:

semiocclusive

Vehicle:

water

Details on dermal exposure:

The test item solution was administered to about 50 cm2 (corresponds to at least 10 % of the body surface area) of the previously clipped skin (dorsal and dorsolateral parts of the trunk); following 24 h incubation with the test item, the dressing was removed and the application site rinsed with warm water

Duration of exposure:

24 h

Doses:

female: 400, 1000 and 2000 mg/kg
male: 400 and 1000 mg/kg

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

animals were observed for 14 days

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: no mortality occurred

Sex:

male

Dose descriptor:

LD50

Effect level:

> 1 000 mg/kg bw

Remarks on result:

other: no mortality occurred

Mortality:

No mortalities occured at any dose level

Clinical signs:

other: No systemic signs of toxicity were noted in all animals.

Gross pathology:

Local effects observed in the 400, 1000 and 2000 mg/kg dose group comprised very slight to severe erythema, very slight or slight edema, scaling, crust formation, bleeding and eczematous skin changes. Additionally, necrosis was observed in the 2000 mg/kg dose group. Due to severe local effects in the female animals of the 2000 mg/kg dose group no further investigation at this dose level was performed in male animals.

Other findings:

Necropsy findings of 4 female animals of the 2000 mg/kg dose group and 1 female animal of the 1000 mg/kg dose group sacrificed at the end of the study comprised several focal skin lesions in the region of the application site, crust formation at the surface and slight focal erythema. No abnormalities were observed in all other animals sacrificed at the end of the study.
The necrotic skin changes in female animals of the 2000 mg/kg dose group assessed by histo-pathological examination indicating full thickness necrosis.

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the conditions of this study the acute dermal median lethal dose (LD50) of the test substance was found to be greater than 2000 mg/kg bw for female rats and greater than 1000 mg/kg bw for male rats. Due to the severe local effects (necrosis) observed at the dose level of 2000 mg/kg bw in the female rats no higher dose levels were assessed in male rats. However, due to lacking systemic effects the acute dermal median lethal dose (LD50) of the test substance is very likely to be greater than 2000 mg/kg bw also in the male animals.

Executive summary:

The study was performed to determine the acute dermal median lethal dose (LD50) of the test substance, applied as a solution in doubly distilled water, in Wistar rats.

The study procedure was based on the EEC, OECD and EPA/OPPTS guidelines.

The test material was applied as a solution in doubly distilled water to the clipped epidermis (dorsal and dorsolateral parts of the trunk) and was covered by a semiocclusive dressing for 24 hours. Dose levels of 400 and 1000 mg/kg bw were applied to groups of 5 male and female rats each. 2000 mgkg bw were given to 5 female rats.

No systemic signs of toxicity were noted in all animals.

The expected body weight gain was generally observed in the course of the study, with the exception of 2 female animals of the 2000 mg/kg bw dose group, which showed stagnation of body weight in the first week of observation.

Local effects observed in the 400, 1000 and 2000 mg/kg bw dose group comprised very slight to severe erythema, vry slight edema, scaling, crust formation, bleeding and eczematoid skin change. In the 2000 mg/kg bw dose group additionally necrosis was observed. Due to severe local effects in the female animals of th 2000 mg/kg bw dose group no further investigation at this dose level was performed in male animals.

No mortality occurred.

Necropsy findings of 4 female animals of the 2000 mg/kg bw dose group and 1 female animal of the 1000 mg/kg bw dose group sacrificed at the end of the study comprised several focal skin lesions in the region of the application site, crust formation at the surface and slight focal erythema. No abnormalities were observed in all other animals sacrificed at the end of the study.

The necrotic skin changes in female animals of the 2000 mg/kg bw dose group assessed by histopathological examination indicating full thickness necrosis.

Under the conditions of this study the acue dermal median lethal dose (LD50) of the test substance was found to be greater than 2000 mg/kg bw for female rats and greater than 1000 mg/kg bw for male rats. Due to the severe local effects (necrosis) observed at the dose level of 2000 mg/kg bw in the female rats no higher dose levels were assessed in male rats. However, due to lacking systemic effects the acute dermal median lethal dose (LD50) of the test substance is very likely to be greater than 2000 mg/kg bw also in the male animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute oral toxicity:

The acute oral toxicity of the test item was assessed similar to OECD401 (BASF AG, 1972). Five rats per dose were given the test item at a 2% aqueous solution once per oral gavage. Animals were observed for seven days and mortality as well clinical signs were monitored. All animals died following application of the two highest doses (i.e. 7200 and 5600 mg/kg), 4/5 males and all female rats at 4500 mg/kg, 2/5 males and 3/5 females at 3600 mg/kg, 0/5 males and 1/5 females at 2800 mg/kg and 1/5 males and 0/5 females at 2250 mg/kg. No clinical signs were noted in the rats; Findings at gross pathology included dilatation of the heart, stomach filled with liquid and dilated, extensive hemorrhagic erosions as well as sanguineous, diarrheic stomach content. Based on these results, the oral LD50 was calculated as 3450 mg/kg for males and females.

Acute inhalation toxicity:

The acute inhalation toxicity of the test item was assessed similar to OECD403. The test substance was applied in a constant quantity with the help of an infusion pump to a 2-step blast pipe. Synthetic air (2.6 bar) was used to generate an aerosol, which was infused into the system. Sprague-Dawley rats were exposed for 4 hrs via the nose/head-only exposure method. Analytic concentrations were 3.72, 1.76, 0.81, 0.17 mg/l. Animals were observed up to 14 days following exposure and clinical signs and symptoms monitored regularly. 9/10 males and 10/10 females died when exposed to 3.72 mg/l, 6/10 males and females at 1.76 mg/l, 0/10 males and 3/10 females at 0.81 mg/l and no animals at 0.17 mg/l. Clinical signs at 3.72 and 1.76 mg/l included reddish head, dyspnea, scrubby, shaggy fur and tremor; at 0.81 mg/l accelerated respiration. No clinical signs were noted at 0.17 mg/L. The only male animal surviving exposure to 3.75 mg/l showed a significantly reduced body weight; furthermore, males at 1.76 mg/l showed some reduction on body weight, whereas all other animals exhibited body weights similar to control animals 14 days subsequent to exposure. Animals that died suffered from acute dilatation of the right heart chamber and strong hyperemia of the lung; furthermore, some animals showed tubule necrosis of the kidney. The clinical effects and the pathological findings in the lung can be attributed to the local toxicity caused by the corrosiveness of the test substance. The other pathological findings are considered to be secondary to the observed lung damage. Using the statistical method described by Finley et al. (1971), the LC50 was calculated as 1.5 mg/L.

In two older studies (BASF AG 1972; BASF AG, 1979) six rats per sex were exposed to a saturated atmosphere of the test substance for 8 hours (inhalation risk test). Mortality did not occur. Accelerated breathing and irritation of the mucosa were observed as clinical signs.

Acute dermal toxicity:

The acute dermal toxicity of the test item was assessed according to OECD402. The test item solution was administered to about 50 cm2 (corresponds to at least 10 % of the body surface area) of the previously clipped skin (dorsal and dorsolateral parts of the trunk) of Wistar rats as semi-occlusive coverage. 24 h later, the dressing was removed and the application site rinsed with warm water to remove residual amounts of the test item. Animals were observed for 14 days. No mortalities occurred at any dose level (up to 2000 mg/kg for females and 1000 mg/kg males). Local effects observed in the 400, 1000 and 2000 mg/kg dose group (females) comprised very slight to severe erythema, very slight or slight edema, scaling, crust formation, bleeding and eczematous skin changes. Additionally, necrosis was observed in the 2000 mg/kg dose group. Due to severe local effects in the female animals of the 2000 mg/kg dose group no further investigation at this dose level was performed in male animals. No systemic clinical signs of toxicity were noted in all animals. Necropsy findings of 4 females of the 2000 mg/kg dose group and 1 female of the 1000 mg/kg dose group sacrificed at the end of the study comprised several focal skin lesions in the region of the application site, crust formation at the surface and slight focal erythema. No abnormalities were observed in all other animals sacrificed at the end of the study. Taken together, the test item does not depict any risk of systemic dermal toxicity and the LD50 is > 1000 mg/kg in males (highest dose tested) and > 2000 mg/kg in females.

There are two older acute dermal toxicity studies available (one in rats, one in rabbits; BASF AG, 1978 and BASF AG, 1979). However, these studies were disregarded for assessment due to methodological deficiencies.

Acute intraperitoneal toxicity:

In an acute toxicity study (BASF AG, 1972) the undiluted test substance was administered to 5 mice per sex via intraperitoneal injection at dose levels of 100, 125, 160, 200, 250, 320 and 400 µL/kg bw. Observation period was until day 14 following treatment. All animals of the highest four dose groups died. Clinical signs were observed like apathy, dyspnoe, and tremor. Gross pathology revealed hyperaemia in the liver and intraabdominal adhesions. Based on the mortality incidences of 0/10, 5/10, 4/10, 10/10, 10/10, 10/10, and 10/10 noted at 100, 125, 160, 200, 250, 320 and 400 µL/kg bw, respectively, the LD50 for the i.p. route of exposure was considered to be approximately 125 µL/kg bw (corresponding to approx. 120 mg/kg bw based on the test substance density of 0.96 g/cm³).

Justification for classification or non-classification

According to Regulation (EC) No 1272/2008 (CLP), the registered substance is classified as acute inhalation toxic category 4 and labelled with "Warning" and H332 "harmful if inhaled".