N'-(3-aminopropyl)-N,N-dimethylpropane-1,3-diamine

Endpoint summary

Description of key information

No repeated dose toxicity study is available on N'-(3-aminopropyl) -N, N-dimethylpropane-1,3-diamine. Its potential toxicity is evaluated by read across from repeated dose toxicity studies performed on the analogue substances, diethylenetriamine (CAS no. 111-40-0) and its dihydrochloride salt (CAS no. 3488-90-2) and dipropylenetriamine (CAS no. 56-18-8). By oral route, the 13-week NOAEL was 41 mg/kg bw/d for diethylenetriamine in rats, and the NOAEL for the F0 parental animals was found to be 15 mg/kg bw/d in an OECD 422 study performed on dipropylenetriamine. By inhalation exposure the 3-week NOAEL was 550 mg/m3 for diethylenetriamine in rats. By dermal route, the chronic NOAEL was 56.3 mg/kg bw three times a week in mice for diethylenetriamine.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories (Kingston, NY, USA)
- Age at study initiation: 6 weeks
- Weight at study initiation: body weight ranges of the males and females at first dose were 105.2-145.5g and 92.1-108.8g, respective! y.
- Fasting period before study: none
- Housing: one per side of divided stainless-steel cages
- Diet: ground, certified Rodent chow No. 5002 (Ralston Purina, St Louis, MO, USA) ad libitum
- Water: ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature: 66-77°F
- Humidity (%): 40-70
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: feed

Vehicle:

other: rat chow

Details on oral exposure:

DIET PREPARATION
Diets were prepared once weekly by mixlng rat chow with DETA.2HCI in a grinder for 30 min to produce a 30,000 ppm premix. Diets of various DETA.2HCl concentrations were prepared by diluting and further blending of this premix with the appropriate amounts of ground chow.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Experimental diets were analysed using HPLC. Homogeneity and stability of the diets were established prior to the start of the study. Homogeneity studies were performed on nine samples (three each from the top, middle and bottom of the mixing bowl) from each diet concentration. The range of the analyses was 89.4 to 109.5% of nominal. Stability studies indicated that DETA.2HCI remained stable for at !east 7 days and 21 days when stored at room temperature in open glass feeder jars and in closed polyethylene containers, respectively. Concentrations were verified for ali diets each week for the first 4 wk of the study and, in subsequent weeks, concentrations concentrations were verified for one diet selected sequentially. The analytical concentrations ranged from 90.6 to 108.6% of nominal.

Duration of treatment / exposure:

90 days

Frequency of treatment:

Ad libitum

Remarks:

Doses / Concentrations:
1000, 7500 and 15000 ppm DETA, HCl
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
70, 530 and 1060 mg DETA, HCl/kg bw/d (41, 307 and 614 mg DETA/kg bw/d) for the males
Basis:
actual ingested

Remarks:

Doses / Concentrations:
80, 620 and 1210 mg DETA, HCl/kg bw/day (46, 360 and 701 mg DETA/kg bw/d) for the females
Basis:
actual ingested

No. of animals per sex per dose:

10

Control animals:

yes, concurrent no treatment

Details on study design:

- Dose selection rationale: A preliminary study with 10 rats per sex per group fed diets containing DET A.2HCI at concentrations of 0, 5000, 10,000, 25,000 or 50,000 ppm for 14 days was conducted to aid in the selection of doses for the present study. There was no mortality in any group, but significant toxicity was observed in the two highest dose groups. Signs of toxicity included decreases in food consumption, body weight and weight gain, and organ weights (liver,
kidneys, spleen, heart and lungs). At 10,000 ppm, decreases in body weight gain, and in Jung and liver weights were noted in males only. The only effect noted at 5000 ppm was decreased Jung weight in males.
- Post-exposure recovery period in satellite groups: the high dose and control groups contained an additional 10 rats per sex that were designated for a 4-wk recovery period.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily for mortality, once daily for overt clinical signs

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): weekly
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to dosing and final killing
- Dose groups that were examined: all

HAEMATOLOGY: Yes
- Time schedule for collection of blood: before dosing, and in wk 6 and 13 of dosing
- Anaesthetic used for blood collection: Yes (methoxyflurane)
- Animals fasted: Yes, about 22 hr
- How many animals: 10 per sex per group + recovery animals
- Parameters: Leucocyte count, Erythrocyte count, Haemoglobin concentration, Haematocrit, Mean corpuscular volume (MCV), Mean corpuscular haemoglobin (MCH), Mean corpuscular haemoglobin concentration, Platelet count, Differential leucocyte count, Reticulocyte count, Prothrombin time, Partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: before dosing, and in wk 6 and 13 of dosing
- Anaesthetic used for blood collection: Yes (methoxyflurane)
- Animals fasted: Yes, about 22 hr
- How many animals: 10 per sex per group + recovery animals
- Parameters : Glucose, Urea nitrogen, Creatinine, Total protein, Albumin, Globulin, Total bilirubin, Direct bilirubin, Indirect bilirubin, Cholesterol, Creatinine kinase, Aspartate aminotransferase, Alanine aminotransferase, Lactic dehydrogenase, Alkaline phosphatase, Calcium, Phosphorus, Sodium, Potassium, Chloride

URINALYSIS: Yes
- Time schedule for collection of urine: for 24 hr from 10 rats per sex per group in wk 6 and 13.
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters: Volume, Specific gravity, pH, Protein, Glucose, Ketone, Bilirubin, Blood, Urobilinogen, Microscopic elements

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

After l3 and 17 (recovery groups) wk of the study, rats were weighed, anaesthetized with methoxyflurane and killed by severing the brachial vessels to permit exsanguination. A complete autopsy was performed.

GROSS PATHOLOGY: Yes

ORGAN WEIGHTS: Yes
brain, liver, kidneys, lungs, spleen, heart, adrenals, thyroid and gonads

HISTOPATHOLOGY: Yes
A histological examination covering an extensive range of tissues was performed on control and high dose (15,000 ppm) animals killed in wk 13. The lungs, liver, kidneys and gross lesions only were examined for the 1000 and 7500 ppm groups, as weil as the controls and recovery groups killed at wk 17

Statistics:

Data for continuous, parametric variables were compared with the control group by the Levene's test for homogeneity of variances, by analysis of variance, and by pooled variance t-tests. The t-tests were used, if the analysis of variance was significant, to delineate which groups differed from the control group. If Levene's test indicated heterogeneous variances, the groups were compared by the analysis of variance of unequal variances, followed if necessary, by separate variance t-tests. Non-parametric data were analysed by the Kruskal-Wallis test or by the Wilcoxon rank sum test as modified by Mann-Whitney. Frequency data were compared using Fisher's exact test where appropriate. The fiducial limit of P < 0.05 (twotailed) was used as the critical leve! of significance for ali tests.

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

CLINICAL SIGNS AND MORTALITY
A single female in the 1000 ppm group died in the wk 8. The cause of death was not apparent from the gross and microscopie examination.
There were no other clinical signs related to DETA.2HCl treatment in all groups over the 90-day exposure period.

BODY WEIGHT AND WEIGHT GAIN
Body weight and body weight gain were reduced compared with controls in the 15,000 ppm males and females throughout the study. The male rats showed only minimal recovery, and their body weights and body weight gains remained lower than controls through wk 17. In contrast, body weights and weight gains of the female rats, consistent with their increase in food consumption during the recovery period, did not differ from those of the controls at the end of the recovery period. At 7500 ppm, body weights were reduced in wk 2, 3, 6 and 9-13 in males and wk 2-6, 8-9 and 13 in females. Body weight gains were reduced throughout the study in males, and in the wk 1-6, 8-10 and 13 in females. There were no treatment-related changes in body weight or body weight gain at 1000 ppm in either sex.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Statistica!ly significant reductions in food consumption compared with controls were observed in the 15,000 ppm males in wk 1-4, 6-7, 9-10 and 12-13, and in the 15,000 ppm females in wk 1-5. A statistically significant decrease was observed in the 7500 ppm males in wk 1 and in the 1000 ppm females in wk 5-6. During the recovery phase (wk 14-17), food consumption was increased relative to control in the females but not the males.
On the basis of the food consumption and body weight data, the dietary concentrations of 1000, 7500 and 15,000 ppm corresponded to mean doses of 70, 530 and 1060 mg/kg/day for the males and 80, 620 and 1210 mg/kg/day for the females.

OPHTHALMOSCOPIC EXAMINATION
Prestudy ophthalmic examinations indicated that all but seven of the rats had minimal to mild corneal dystrophy. Because of the historically high incidence of this lesion for this strain of rats from all animal suppliers, and the lack of potential impact on interpreting the toxicity of DETA.2HCI, these animals were accepted for study despite the presence of this corneal lesion. In the terminal ophthalmic examination, corneal dystrophy was diagnosed in all but one animal. The lesions were again mild and occurred with equal frequency across all groups. No other ocular lesions were observed.

HAEMATOLOGY
At wk 6, increases in mean corpuscular volume (MCV) in the 7500 and 15,000 ppm males and increases in mean corpuscular haemoglobin (MCH) in the 15,000 ppm males were observed. The MCV and MCH in the 7500 and 15,000 ppm males at wk 13 and in the 15,000 ppm males at wk 17 (recovery) were elevated. In all cases, the magnitude of these changes was 3% or less compared with control. These changes were related to statistically significant increases in reticulocytes, observed at wk 6 and 13. Increases in MCV were also observed in the 7500 and 15,000 ppm females at wk 6, and in the 15,000 ppm females at wk 17.
A statisticaliy significant increase in total leucocyte count, attributed to a statisticaliy significant increase in lymphocytes, was observed in the 7500 ppm and 15,000 ppm females at wk 6 and 13. At wk 13, statisticaliy significant increases in segmented neutrophils also contributed to the increase
in total leucocyte numbers. Although total leucocytes were no longer significantly altered following a 4-wk recovery, the lymphocyte increase was still
statisticaliy significant.

CLINICAL CHEMISTRY
The serum glucose concentrations were depressed in females exposed to 7500 and 15,000 ppm at wk 13, and persisted foliowing a 4-wk recovery period.

URINALYSIS
Urine pH was significantly increased in the 7500 and 15,000 ppm groups at wk 6 and 13. This alteration, however, was no longer observed foliowing the recovery period.

ORGAN WEIGHTS
Statisticaliy significant increases in absolute kidney and brain weights and in relative kidney, brain and testes weights were observed in the males at
15,000 ppm. The relative kidney and testes weights remained elevated foliowing the 4-wk recovery period. The females showed a significant, dose-related increase in relative kidney, brain, and Iiver weights at 7500 and 15,000 ppm, and in relative heart and adrenal weights at 15,000 ppm. Following
the recovery period, only the relative Jiver and adrenal weights remained elevated.

GROSS PATHOLOGY
One female rat from the 1000 ppm group died on study day 55. This rat was found to be emaciated at autopsy, but the specifie cause of death could not be determined. There were no treatment-related gross findings in any group.

HISTOPATHOLOGY: NON-NEOPLASTIC
There were no statistically significant differences in the incidences of microscopic findings between the 15,000 ppm and control groups killed at wk 13 or 17.

Dose descriptor:

NOAEL

Effect level:

1 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: depressed body weights and minor alterations in clinical pathology measurements and organ weights. NOAEL corresponding to 41 mg DETA/kg bw/d in male rats and 46 mg DETA/kg bw/d in female rats

Critical effects observed:

not specified

Conclusions:

In summary, dietary exposure to DETA.2HCI at 7500 and 15,000 ppm for 90 consecutive days resulted in systemic toxicity, as indicated by depressed
body weights and minor alterations in clinical pathology measurements and organ weights. A no-observed-effect leve! of 1000 ppm (corresponding
to 41 mg/kg/day in male rats and 46 mg DETA/kg/day in female rats) was established in this study.

Executive summary:

Fischer 344 rats were fed a diet containing the dihydrochloride salt of diethylenetriamine (DETA, 2HCl) at concentrations of 1000, 7500 or 15,000 ppm for 90 consecutive days. Based on food consumption and body weight, the mean corresponding dosages were 70, 530 and 1060 mg/kg/day for the males and 80, 620 and 1210 mg/kg/day for the females. Decreases in food consumption were observed intermittently throughout the dosing period in both sexes at 15,000 ppm. Dose-related decreases in body weight or weight gain were observed for both sexes at 7500 and 15,000 ppm. Changes in clinical pathology measurements observed at 7500 and 15,000 ppm included increases in mean corpuscular volume and mean corpuscular haemoglobin in males, and increases in mean corpuscular volume, total leucocytes and urinary pH, and a decrease in serum glucose concentration in females. The relative kidney, brain and testes weights were increased in the 15,000 ppm males. In the females, the relative kidney, brain and liver weights were increased at 7500 and 15,000 ppm, and the relative heart and adrenal weights were elevated at 15,000 ppm. There were no treatment-related clinicat signs, gross autopsy or histopathological findings for either sex at any dose level. Animals improved only slightly from the effects of treatment following a 4-wk recovery period. A no-observed-effect leve! of 1000 ppm (corresponding

to 41 mg DETA/kg/day in male rats and 46 mg DETA/kg/day in female rats) was established in this study.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

41 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Read across to diethylenetriamine

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: inhalation

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

no guideline followed

Principles of method if other than guideline:

Rats were exposed 6 hours/day for 15 exposures.

GLP compliance:

no

Limit test:

no

Species:

rat

Strain:

other: Alderley Park specific-pathogen-free

Details on test animals or test system and environmental conditions:

Average weight 200 grams

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

clean air

Remarks on MMAD:

MMAD / GSD: Not applicable

Details on inhalation exposure:

Design of exposure chambers
In all of these experiments the animals have been exposed to dynamic atmospheres, that is, to atmospheres continuously generated and passed through the exposure chamber. The design of exposure chamber has varied with the number of animals involved and with the nature of the substance under investigation. For groups of four or fewer rats a glass desiccator, containing wire mesh partitions to separate the animals, was used. Larger numbers, up to eight rats, were exposed in the chamber described elsewhere (Gage, 1959); usually the inner Perspex chamber of that design was replaced by a glass cylinder, 30 cm diameter and 25 cm high. For atmospheres containing particulate matter a chamber with a more pyramidal top (Gage, 1968) was used.

Generation of the test atmospheres
The air used foi the atmospheres was filtered, dried to a relative humidity of less than 10%, and supplied at a line pressure of 1 atm (1.013 x 10 E5 Nm-2). A nearly saturated vapour obtained by passing air through a liquid contained in a bubbler with a sintered glass air-distributor disc. The volume of the liquid was usually 10-20 ml and, if the size of the sample available permitted, it was replaced daily. Unless otherwise stated, the bubbler was maintained in a water-bath at room temperature, about 20°C.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The nearly saturated concentration was estimated by weighing the sample before and after the day's run, and telating the weight loss to the volume of air passing. This concentration, expressed in milligrammes per litre, was converted to parts per million on the assumption that the sample was pure. Both of these estimates are only approximate.

Duration of treatment / exposure:

3 weeks

Frequency of treatment:

6-hour/day, 5 days/week

Remarks:

Doses / Concentrations:
550 mg/m3 (130 ppm)
Basis:
analytical conc.

No. of animals per sex per dose:

2

Control animals:

not specified

Details on study design:

- Dose selection rationale: In the initial experiments the concentrations were selected to produce, if possible, acute effects after short exposures.

If at any stage effects were observed which could be attiributed to the exposure, the experiment was repeated with progressively lower concentrations until a concentration was reached which was without effects on the animals. At intervals of about two months, batches of control rats were maintained in a chamber for the exposure period, in order to check the characteristics of the colony.

Observations and examinations performed and frequency:

The rats were weighed each morning, and their conditions and behaviour were recorded throughout the exposure period. Urine was collected overnight after the last exposure day for biochemical tests. The animals were left overnight with food and drink. On the following day the rats were anaesthetized with halothane and partially exsanguinated by heart puncture for haematological tests.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

After a gross examination of the organs, the lungs were inflated with formol-saline and immersed in the same fixative. The following organs were also taken for microscopical examination after fixation in formol-corrosive: lungs, liver, kidneys, spleen, and adrenals; and occasionally heart, jejunum, ileum, and thymus.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

not specified

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

Details on results:

DETA at 15 X 6-hour exposures produced no toxic signs (hair coarsened). The autopsy and all organs were normal.

Dose descriptor:

NOAEC

Effect level:

550 mg/m³ air (analytical)

Sex:

male/female

Basis for effect level:

other: no toxic signs (hair coarsened): autopsy, organs normal

Critical effects observed:

not specified

The effects of inhaled DETA were examined in a subacute study. Groups of rats were exposed to an essentially saturated vapor of DETA for 6 hrs/day for 15 exposures. There were no effects based on urinalysis or hematological parameters and gross or histopathological examinations.

Executive summary:

ln a subacute inhalation study, 2 male and 2 female rats (Alderley Park, SPF, 200 g) inhaled diethylenetriamine for 6 hours/ day, 5 days/week for 15 days. The diethylenetriamine-containing atmosphere was produced by passing air through about 10 to 20 ml of the product (which was renewed every day) in a glass inhalation chamber. The calculated concentration of the substance in the chamber was 0.55 mg/1 (130 ppm). The behaviour of the rats and their weight was recorded every day. The rats had ruffled fur but no other signs of intoxication were observed during the entire study. At the end of the study, the animals were killed and post-mortems carried out. The lungs, liver, kidneys, spleen and adrenal glands were examined histologically, but no effects on the organs were found.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEC

550 mg/m³

Study duration:

subacute

Species:

rat

Quality of whole database:

Read across to diethylenetriamine

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records

Reference

Endpoint:

chronic toxicity: dermal

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

Mice were dosed dermally three times/week for a lifetime. Gross and histopathological examinations were performed on all animals.

GLP compliance:

not specified

Species:

mouse

Strain:

other: C3H/HeJ

Sex:

male

Type of coverage:

open

Vehicle:

water

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

complete life span (ca. 20 months)

Frequency of treatment:

3 times weekly

Remarks:

Doses / Concentrations:
1.25 mg/animal
Basis:
other: nominal in water 25 µl in 5% solution v/v

Remarks:

Doses / Concentrations:
56.3 mg/kg bw
Basis:
other: Estimate as calculated from data from average of initial body weights and milligrams administered

Control animals:

yes, concurrent vehicle

Details on results:

See section 7.7

Dose descriptor:

NOAEL

Effect level:

>= 56.3 mg/kg bw/day

Sex:

male

Basis for effect level:

other: 3 times weekly

Critical effects observed:

not specified

Executive summary:

Diethylenetriamine (DETA) was tested in a total of 100 male C3H/HeJ mice in a study involving dermal application of two products of different degrees of purity, DETA High Purity (DETA-HP, 96.77% diethylenetriamine, 1.78% N-(2-aminoethyl)piperazine, remaining components other amines) and DETA Commercial Grade (DETA-C 90.8% diethylenetriamine, 8.9% N-(2-aminoethyl)piperazine, 0.34% ethylenediamine). The test substances were applied as a 5% aqueous solution. A 10% diethylenetriamine solution had caused open skin wounds, and so the non-irritating 5% solution was considered to be the highest possible concentration that could be used in a lifetime study. The treatment involved the application of 25 µl of the test solution (1.25 mg) to the dorsal skin of the mice (which was shaved once a week) three times a week for their entire lives. The mice were observed daily. After 18 months, the skin, liver, kidneys and lungs from 10 mice/group were fixed for histological examination. The average survival time for the mice was 587 days in the DETA-HP group and 662 days in the DETA-C group. The animals in the control group, which were treated with 25 µl water, lived for 626 days on average. 2 tumours were found in total. One mouse in the control group had a sebaceous gland adenoma of the thorax, and one in the DETA-C group had a cavernous haemangioma of the neck region. This type of haemangioma was also observed in historical controls in the institute carrying out the study. Also the effects on other organs corresponded to the spontaneous occurrence in the control groups of the strain studied. Diethylenetriamine thus had no carcinogenic activity in this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

56.3 mg/kg bw/day

Study duration:

chronic

Species:

mouse

Quality of whole database:

Read across to diethylenetriamine

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

chronic toxicity: dermal

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study well documented, meets generally accepted scientific principles, acceptable for assessment

Reason / purpose for cross-reference:

reference to same study

Principles of method if other than guideline:

Mice were dosed dermally three times/week for a lifetime. Gross and histopathological examinations were performed on all animals.

GLP compliance:

not specified

Species:

mouse

Strain:

other: C3H/HeJ

Sex:

male

Type of coverage:

open

Vehicle:

water

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

complete life span (ca. 20 months)

Frequency of treatment:

3 times weekly

Remarks:

Doses / Concentrations:
1.25 mg/animal
Basis:
other: nominal in water 25 µl in 5% solution v/v

Remarks:

Doses / Concentrations:
56.3 mg/kg bw
Basis:
other: Estimate as calculated from data from average of initial body weights and milligrams administered

Control animals:

yes, concurrent vehicle

Details on results:

See section 7.7

Dose descriptor:

NOAEL

Effect level:

>= 56.3 mg/kg bw/day

Sex:

male

Basis for effect level:

other: 3 times weekly

Critical effects observed:

not specified

Executive summary:

Diethylenetriamine (DETA) was tested in a total of 100 male C3H/HeJ mice in a study involving dermal application of two products of different degrees of purity, DETA High Purity (DETA-HP, 96.77% diethylenetriamine, 1.78% N-(2-aminoethyl)piperazine, remaining components other amines) and DETA Commercial Grade (DETA-C 90.8% diethylenetriamine, 8.9% N-(2-aminoethyl)piperazine, 0.34% ethylenediamine). The test substances were applied as a 5% aqueous solution. A 10% diethylenetriamine solution had caused open skin wounds, and so the non-irritating 5% solution was considered to be the highest possible concentration that could be used in a lifetime study. The treatment involved the application of 25 µl of the test solution (1.25 mg) to the dorsal skin of the mice (which was shaved once a week) three times a week for their entire lives. The mice were observed daily. After 18 months, the skin, liver, kidneys and lungs from 10 mice/group were fixed for histological examination. The average survival time for the mice was 587 days in the DETA-HP group and 662 days in the DETA-C group. The animals in the control group, which were treated with 25 µl water, lived for 626 days on average. 2 tumours were found in total. One mouse in the control group had a sebaceous gland adenoma of the thorax, and one in the DETA-C group had a cavernous haemangioma of the neck region. This type of haemangioma was also observed in historical controls in the institute carrying out the study. Also the effects on other organs corresponded to the spontaneous occurrence in the control groups of the strain studied. Diethylenetriamine thus had no carcinogenic activity in this study.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Study duration:

chronic

Species:

mouse

Quality of whole database:

Read across to diethylenetriamine

Additional information

No repeated dose toxicity study is available on N'-(3-aminopropyl) -N, N-dimethylpropane-1,3-diamine. Its potential toxicity is evaluated by read across from repeated dose toxicity studies performed on the analogue substances, diethylenetriamine (CAS no. 111-40-0) and its dihydrochloride salt (CAS no. 3488-90-2) and dipropylenetriamine (CAS no. 56-18-8).

Oral route

Fischer 344 rats were fed a diet containing the dihydrochloride salt of diethylenetriamine (DETA.2HCl) at concentrations of 1000, 7500 or 15,000 ppm for 90 consecutive days (Leung and van Miller, 1997). Based on food consumption and body weight, the mean corresponding dosages were 70, 530 and 1060 mg/kg/day for the males and 80, 620 and 1210 mg/kg/day for the females. Decreases in food consumption were observed intermittently throughout the dosing period in both sexes at 15,000 ppm. Dose-related decreases in body weight or weight gain were observed for both sexes at 7500 and 15,000 ppm. Changes in clinical pathology measurements observed at 7500 and 15,000 ppm included increases in mean corpuscular volume and mean corpuscular haemoglobin in males, and increases in mean corpuscular volume, total leucocytes and urinary pH, and a decrease in serum glucose concentration in females. The relative kidney, brain and testes weights were increased in the 15,000 ppm males. In the females, the relative kidney, brain and liver weights were increased at 7500 and 15,000 ppm, and the relative heart and adrenal weights were elevated at 15,000 ppm. There were no treatment-related clinical signs, gross autopsy or histopathological findings for either sex at any dose level. Animals improved only slightly from the effects of treatment following a 4-wk recovery period. A no-observed-effect level of 1000 ppm (corresponding to 41 mg DETA/kg/day in male rats and 47 mg DETA/kg/day in female rats) was established in this study.

In an OECD 422 study (BASF, 2012), the test substance Dipropylenetriamine was administered as an aqueous solution at 0, 5, 15 and 50 mg/kg bw/d daily by gavage to groups of 10 male and 10 female Wistar rats to test for repeated dose toxicity and screen for potential reproductive and developmental effects. After a two-week premating period, these parental animals were mated and the females were allowed to give birth and suckle the offspring until sacrifice on PND 4. Males were treated for 30 days, females for 49 days. Food consumption and body weights of F0 parents were determined. Clinico-chemical and hematological examinations as well as urinalyses were performed in 5 animals per sex and group towards the end of the administration period. At the end of the administration period a FOB was performed and motor activity was measured in 5 parental males and females per group. After sacrifice all F0 parental animals were assessed by gross pathology. Weights of selected organs were recorded and a histopathological examination was performed.

Food consumption was mildly reduced in the parental females of the high-dose group (50 mg/kg bw/d) during late-gestation (GD 14-20). This lower food consumption corresponded to slightly decreased parental female body weights beginning during this same time period, but only becoming statistically significant on PND 7. Similarly, there was a statistically significant decrease in body weight gain between GD 14 and GD 20. While there does seem to be an overall downward trend in the body weight parameters of the rats of the high dose group that may indicate the beginnings of systemic maternal toxicity, this reduction was small (<8%). This trend is only a borderline case; however, as toxicity cannot be excluded, it is considered adverse. No alterations to either parental female or male body weight parameters were observed during the premating or mating phases. No changes in either food consumption or body weight parameters were noted at either the low- (5 mg/kg bw/d) or mid-doses (15 mg/kg bw/d). Regarding clinical pathology, a slight lymphopenia was noted in males of the high-dose group (50 mg/kg bw/d). A reason for this alteration could not be found. No deviations from normal were observed in parental females. Regarding pathology, the thymus of 9 out of 10 males (dose group 50 mg/kg bw/d) showed minimal to severe decreased cellularity in the cortex and medulla. This change, which is regarded as treatment-related and adverse was associated with significantly decreased mean absolute and relative thymus weight and reduced organ size in 5 of 10 male animals. A minimal but significant increase of the mean relative liver weight in males of the high dose group 3 (50 mg/kg bw/d) occurred without any histopathological correlate. Therefore, this change is regarded as treatment-related but not adverse. No alterations to reproductive performance or fertility were identified at any dose in the parental rats..

Thus, under the conditions of this study, the NOAEL for general, systemic toxicity of the test substance is 15 mg/kg bw/d for the F0 parental males based on the lymphopenia and/or thymus pathology identified at 50 mg/kg bw/d. The NOAEL for F0 parental females was found to be 15 mg/kg bw/d based on the changes to body weight parameters at 50 mg/kg bw/d.

Inhalation exposure

In a subacute inhalation study, 2 male and 2 female rats (, SPF, 200 g) inhaled diethylenetriamine for 6 hours/ day, 5 days/week for 15 days (Gage, 1970). The diethylenetriamine-containing atmosphere was produced by passing air through about 10 to 20 ml of the product (which was renewed every day) in a glass inhalation chamber. The calculated concentration of the substance in the chamber was 0.55 mg/1 (130 ppm). The behaviour of the rats and their weight was recorded every day. The rats had ruffled fur but no other signs of intoxication were observed during the entire study. At the end of the study, the animals were killed and post-mortems carried out. The lungs, liver, kidneys, spleen and adrenal glands were examined histologically, but no effects on the organs were found.

Dermal route

Diethylenetriamine (DETA) was tested in a total of 100 male C3H/HeJ mice in a study involving dermal application of two products of different degrees of purity, DETA High Purity (DETA-HP, 96.77% diethylenetriamine, 1.78% N-(2-aminoethyl) piperazine, remaining components other amines) and DETA Commercial Grade (DETA-C 90.8% diethylenetriamine, 8.9% N-(2-aminoethyl) piperazine, 0.34% ethylenediamine) (de Pass et al., 1987). The test substances were applied as a 5% aqueous solution. A 10% diethylenetriamine solution had caused open skin wounds, and so the non-irritating 5% solution was considered to be the highest possible concentration that could be used in a lifetime study. The treatment involved the application of 25 µl of the test solution (1.25 mg/mouse equivalent to 56.3 mg/kg bw) to the dorsal skin of the mice (which was shaved once a week) three times a week for their entire lives. The mice were observed daily. After 18 months, the skin, liver, kidneys and lungs from 10 mice/group were fixed for histological examination. The average survival time for the mice was 587 days in the DETA-HP group and 662 days in the DETA-C group. The animals in the control group, which were treated with 25 µl water, lived for 626 days on average. 2 tumours were found in total. One mouse in the control group had a sebaceous gland adenoma of the thorax, and one in the DETA-C group had a cavernous haemangioma of the neck region. This type of haemangioma was also observed in historical controls in the institute carrying out the study. Also the effects on other organs corresponded to the spontaneous occurrence in the control groups of the strain studied. Diethylenetriamine thus had no carcinogenic activity in this study.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Key study

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
Key study

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
Key study

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
Key study

Justification for classification or non-classification

According to REGULATION (EC) No 1272-2008 and Annex VI of Commission Directive 2001/59/EC:

Not classified, according to the data available on the analogue substances, diethylenetriamine and dipropylenetriamine.