Registration Dossier

Administrative data

Description of key information

Trimethylenediamine is structurally very similar to ethylenediamine (difference: chain length / one CH2-group) or ethylenediammonium dichloride (applied doses of ammonium and chloride of no toxicological relevance). Thus, repeated oral dose studies' results generated by these substances are justified for read across to assess the repeated doses toxicity of trimethylenediamine.
Results:
- Oral route: For repeated oral toxicity a NOAEL of 27.9 mg/kg bw /day trimethylenediamine could be derived by read across from a respective study with ethylenediammonium dichloride.
- Inhalation: An available NOAEC for EDA (analytical) could be calculated to be 177.6 mg/m³ air trimethylenediamine.
- Dermal route: There were no treatment-related macroscopic or histopathologic findings in a lifetime dermal carcinogenicity study with EDA at the highest possible concentration in mice. Thus, respective results are also expected for trimethylenediamine (read across). A NOAEL of 10.2 mg/kg bw /day trimethylenediamine was derived due to the maximum tolerated dose (for local effects).

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:
experimental study
Adequacy of study:
key study
Study period:
1983
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was performed pre-GLP. No guideline was available. Trimethylenediamine is structural very similar to ethylenediamine (difference: chain length / one CH2-group) or ethylenediammonium dichloride (doses of ammonium and chloride not toxicological relevant), respectively. Thus, the test result is justified for read across to assess the toxicity of trimethylenediamine.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
not specified
GLP compliance:
no
Remarks:
pre-GLP
Limit test:
no
Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Microbiological Associates, Inc., Walkersville, MD
- Age at study initiation: 41 days
- Weight at study initiation: 101-152 g (males) 91-110 g (females)
- Fasting period before study: no information provided
- Housing: suspended wire-bottom-and-front stainless steel cages. Three males or 5 females were housed in each cage.
- Water: ad libitum
Route of administration:
oral: feed
Vehicle:
other: no data (most likely feed only)
Details on oral exposure:
New concentrations of feed were prepared weekly, with the percentage of EDA.2HCl in the diet adjusted to maintain a constant dosage level in mg/kg for each sex according to the average body weight gain and diet consumption.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
A spectrophotometric method was employed for the analysis of EDA.2HCI in the diet.
Duration of treatment / exposure:
3 months
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:
50 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
260 mg/kg bw/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1040 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose:
10 animals per sex
Control animals:
yes, plain diet
Observations and examinations performed and frequency:
BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- 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

WATER CONSUMPTION : Yes
- Time schedule for examinations: Monthly

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: Immediately before sacrifice
- Parameters c examined: Red and white blood cell counts, measurement of hemoglobin and mean corpuscular volume, calculation of hematocrit, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and determination of differential white blood cell counts

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: Immediately before sacrifice
- Animals fasted: No data
- How many animals:
- Parameters examined: Measurement of serum concentrations of glucose, urea nitrogen, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, total protein, albumin and creatinine.

URINALYSIS: Yes
- Time schedule for collection of urine: One week before sacrifice

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
All rats were given a complete gross necropsy examination and organ weights were recorded for the brain, liver, kidneys, spleen, heart, adrenals and
testes. Tissues were taken and fixed in 10% neutral buffered formalin. All tissues were processed for paraffin embedding, sectioned at 5 microns and stained with hematoxylin and eosin. Microscopic lesions were graded as to severity, where possible, into 4 categories (mild, moderate, marked or severe)
Details on results:
CLINICAL SIGNS AND MORTALITY
No deaths or clinical signs of toxicity.

BODY WEIGHT AND WEIGHT GAIN
Body weight gain was depressed markedly for both sexes at the high dosage level

FOOD AND WATER CONSUMPTION
There was marked reduction in diet consumption in the high level dose females and a significant increase in the low level dose females. While water
consumption rates were equivalent in the male test and control animals, there was a dose-related reduction for the female test animals.

CLINICAL CHEMISTRY
Changes in clinical chemistry values included a reduction in serum glucose level and an elevation of alkaline phosphatase activity, AST and ALT activities

HAEMATOLOGY
In male rats, depression of the red blood cell counts and increased mean corpuscular volumes were seen. In the female rats, in addition to the above changes, there were also depression of hematocrit and hemoglobin values and an increase in mean corpuscular hemoglobin.

URINALYSIS
The median urinary pH at the high dosage level was lowered significantly in both sexes

ORGAN WEIGHTS
In the male rats, there was a significant reduction in liver, kidney, spleen and heart weight and a concomitant decrease in some of the relative organ weights. Similarly, in the females, there was a reduction in liver, heart, adrenal and brain weights with an decrease of relative liver weight.

PATHOLOGY
There was no dose-related gross lesions in any animal on the study. The most significant histologic changes were present in the livers of the high dosage level animals, particularly in the females. The liver changes, termed "hepatocellular pleomorphism", consisted of an increase in the size of hepatocytes and hepatocyte nuclei, increased variation in nuclear size and shape, and an increase in the number of multinucleate hepatocytes. Occasional degenerating hepatocytes were also seen.
Dose descriptor:
NOAEL
Effect level:
ca. 50 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Study result generated with EDA*2HCl as test material.
Dose descriptor:
LOAEL
Effect level:
260 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Study result generated with EDA*2HCl as test material. Clinical chemistry: elevation of alanine aminotransferase activity in males;  Haematology :Increased mean corpuscular volumes in females.
Dose descriptor:
NOAEL
Effect level:
22.6 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
/ recalculated to EDA
Sex:
male/female
Basis for effect level:
other: Recalculation to EDA base from the test results with EDA*2HCl
Dose descriptor:
LOAEL
Effect level:
117 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
/ recalculated to EDA
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Dose descriptor:
NOAEL
Effect level:
27.9 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
/calculated from EDA to trimethylenediamine
Sex:
male/female
Basis for effect level:
other: Study result generated with EDA*2HCl as test material was recalculated to EDA base and then - for read across - calculated for trimethylenediamine.
Dose descriptor:
LOAEL
Effect level:
144 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
/calculated from EDA to trimethylenediamine
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Critical effects observed:
not specified

High dose group: 

Diet and water consumption significantly reduced in the high dose female rats. Significant reduction in body weight gain of both sexes in the high dose group; significant reduction in absolute weights of liver and heart (both sexes), adrenal and brain (female), kidney and spleen (male) in the high dose group; increase of relative weight of brain (both sexes), spleen and heart (female) and testis in the high dose group. Significant elevation of alkaline phosphatase activity in males and females. Significant elevation of alanine aminotransferase activity in males and females of high dose groups. Increased mean corpuscular volumes in males and females. Significant increase of mean corpuscular hemoglobin and significant depression of hematocrit and hemoglobin values; significant depression of red blood cell counts, serum glucose level and urinary pH (from 6.0 to 5.0) and significant elevation of aspartate aminotransferase activity in both sexes of the high dose group; hepatocellular pleomorphism in 7/10 female and 2/10 male (control: 0/10 of each sex) in high dose group, hepatocellular degeneration and significant increased prevalence of tracheitis in male of the high dose group.

Intermediate dose group: 

Water consumption significantly reduced in female rats. Significant elevation of alanine aminotransferase activity in males of intermediate dose groups. Increased mean corpuscular volumes in females.

Low dose group:

 Water consumption significantly reduced in female rats. Significant elevation of alkaline phosphatase activity in males, this effect was not noted in the intermediate dose group.

Endpoint conclusion
Dose descriptor:
NOAEL
27.9 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Non GLP, but scientifically well documented studies.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1954
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
pre-GLP, no guideline available at time of study performance Trimethylenediamine is structurally very similar to the test substance ethylenediamine (difference: chain length / one CH2-group). Thus, the test result is justified for read across with respect to trimethylenediamine.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 412 (Subacute Inhalation Toxicity: 28-Day Study)
Deviations:
not specified
Principles of method if other than guideline:
Four exposure groups of 30 rats (15 female/15 male) were exposed to EDA vapour. Each exposure group had a control group.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Sherman
Sex:
male/female
Route of administration:
inhalation: vapour
Type of inhalation exposure:
whole body
Vehicle:
air
Remarks on MMAD:
MMAD / GSD: Not determined.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Chamber air was passed through an acidified glassbead adsorption tube. The captured Ethylenediamine was quantified by titration.
Duration of treatment / exposure:
6 weeks
Frequency of treatment:
7 h/day, 5 days/week
Remarks:
Doses / Concentrations:
484 ppm
Basis:
analytical conc.
Remarks:
Doses / Concentrations:
225 ppm
Basis:
analytical conc.
Remarks:
Doses / Concentrations:
182 ppm
Basis:
analytical conc.
Remarks:
Doses / Concentrations:
59 ppm
Basis:
analytical conc.
No. of animals per sex per dose:
15 male and 15 female
Control animals:
yes
Clinical signs:
effects observed, treatment-related
Mortality:
mortality observed, treatment-related
Body weight and weight changes:
effects observed, treatment-related
Dose descriptor:
NOAEC
Effect level:
59 ppm (analytical)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: clinical signs; body weight; organ weights
Dose descriptor:
NOAEC
Effect level:
144 mg/m³ air
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: clinical signs; body weight; organ weights (as above)
Remarks on result:
other: calculated from 59 ppm
Dose descriptor:
NOAEL
Effect level:
48 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Recalculated using rat weight 0.425 kg (mean male/female) and inhalation volume 0.33 m3/7h (values from guidance document)
Dose descriptor:
LOAEC
Effect level:
132 ppm (analytical)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
Dose descriptor:
LOAEC
Effect level:
323 mg/m³ air
Based on:
test mat.
Sex:
male/female
Basis for effect level:
mortality
Remarks on result:
other: calculated from 132 ppm
Dose descriptor:
LOAEL
Effect level:
107 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Recalculated using rat weight 0.425 kg (mean male/female) and inhalation volume 0.33 m3/7h (values from guidance document)
Dose descriptor:
NOAEC
Effect level:
177.6 mg/L air (analytical)
Based on:
test mat.
Remarks:
/calculated from EDA to trimethylenediamine
Sex:
male/female
Basis for effect level:
other: /calculated from EDA to trimethylenediamine
Remarks on result:
other: /calculated from EDA to trimethylenediamine
Critical effects observed:
not specified

132 ppm: The death of 4/30 animals was attributed to lung infection (not substance-related); slight depilation; body weight gain and relative weights of liver and kidney were not affected; no substance-related macroscopic or histologic changes.

225 ppm: The death of 16/30 was substance-related and another 10 death were considered not to be substance-related; the 4 rats which survived showed significantly lower weight gain and higher relative weights of liver and kidney; cloudy swelling of the liver and of the loop and convoluted tubules of the kidney; lung congestion was observed in exposed as well as in control rats in similar proportions.
484 ppm: All rats died within 20 days of initial exposure; depilation was first observed on the 6th day of exposure; cloudy swelling of the liver (in 23/28 animals), cloudy swelling and degeneration of convoluted tubules (in 7/28 animals); congestion of the lung (in 17/28 animals) and of the adrenal cortex (in 5/28 animals).
59 ppm: No effect on weight gain or organ weights. No depilation. No significant damage to examined tissues; lung, liver and kidney.

In conclusion.

NOAEC in a subacute inhalation study was 144 mg/m³ air (EDA, analytical), corresponding to a NOAEC of 177.6 mg/m³ air trimethylenediamine (read across).

Endpoint conclusion
Dose descriptor:
NOAEC
177.6 mg/m³
Study duration:
subacute
Species:
rat
Quality of whole database:
Non GLP, but scientifically well documented study.

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: dermal
Remarks:
other: cancerogenicity
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: No data on guideline and GLP. Trimethylenediamine is structural very similar to the test substance ethylenediamine (difference: chain length / one CH2-group). Thus, the test result is justified for read across with respect to trimethylenediamine.
Qualifier:
no guideline followed
Principles of method if other than guideline:
The maximum tolerated dose (for local effects) was applied to the back of 50 male mice three times weekly throughout their lifespan. Positive (3-methylcholanthrene) and negative (water) control groups (50 mice/group) were included.
GLP compliance:
no
Limit test:
no
Species:
mouse
Strain:
C3H
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: C3H/HeJ mice from Jackson Laboratories, Bar Harbor, Maine
- Age at study initiation: 74 to 79 days of age
- Housing: Housed individually in stainless steel cages with wire mesh floors. All mice were housed in the same room with
controlled lighting
- Diet (e.g. ad libitum): Ziegler Bros. NIH 07 pellets (Gardners, Pa.)
- Water (e.g. ad libitum): water from an automatic watering system
Type of coverage:
open
Vehicle:
water
Details on exposure:
Mice were treated three times weekly for their complete life span with 25 µL per application of each substance. Substances were applied with an Eppendorf pipet to the back of each mouse from which the fur was clipped once weekly.

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentrations verified monthly during the study
Duration of treatment / exposure:
complete life span

Frequency of treatment:
3x/wk
Remarks:
Doses / Concentrations:
25 μL of 1% aqueous solution/application
Basis:

No. of animals per sex per dose:
Treatment group singly housed: 50 mice
Control animals:
yes, concurrent vehicle
Details on study design:
Control group singly housed: 50 mice received distilled water. Control group housed 5/cage: 40 mice received water
Positive control:
Positive control group housed 5/cage: 40 mice received 0.1% 3-methylcholanthrene in acetone
Observations and examinations performed and frequency:
All mice were examined daily, and the onset and progress of tumor growth were recorded monthly.
Dose descriptor:
NOAEL
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Mild fibrosis suggesting irritation to skin was noted
Remarks on result:
not measured/tested
Remarks:
25 μL of 1% solution used; Effect level not specified in more detail
Dose descriptor:
NOAEL
Effect level:
8.3 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Weight of male mice 0.03 kg
Dose descriptor:
NOAEL
Effect level:
10.2 mg/kg bw/day (actual dose received)
Based on:
test mat.
Remarks:
/calculated from EDA to trimethylenediamine
Sex:
male
Basis for effect level:
other: calculated from EDA to trimethylenediamine
Critical effects observed:
not specified

Mean survival time of the exposure group for substance 2 (598 days) was shorter than that of the singly housed control group (626 days); no treatment-related macroscopic or histopathologic findings; one mouse of the exposure group had a dermal fibrosis at application site and another one had a mammary adenocarcinoma. One sebaceous adenoma of the skin of the thorax was noted in the control group individually housed.

In the exposure group for substance 1, 1 mouse had a myxosarcoma at the base of the tail, and 11 animals had mild to moderate dermal fibrosis, suggesting skin irritation. Survival time did not differ from negative control groups.

In the 3-methylcholanthrene group, 39 of 40 mice had skin tumors including 37 with confirmed squamous cell carcinomas.

In conclusion:

The dermal repeated dose NOAEL was 8.3 mg/kg bw/day EDA (actually received dose), corresponding to 10.2 mg/kg bw/day trimethylenediamine (read across).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
10.2 mg/kg bw/day
Study duration:
chronic
Species:
mouse
Quality of whole database:
Non GLP, but scientifically well documented study.

Additional information

Repeated dose toxicity: oral

In a three month dietary study (Yang, 1983), male and female rats were fed targeted doses of 0, 50, 250 or 1000 mg/kg/day of EDA-2HCl. There were no deaths and no abnormal clinical signs noted during the study. Body weight gains were significantly decreased in the high dose group which affected a number of absolute and relative organ weights in both males and females. Water consumption was comparable to control values at all dose levels in males but was decreased in a dose-response manner in female rats at all 3 dose levels. Slight reductions in serum glucose levels and an elevation of alkaline phosphatase, AST and ALT activities were observed in the high dose group. An elevation of ALT activity was also observed in the intermediate dose male rats. Urinary pH in the high dose group was decreased in both males and females. There were no dose-related gross lesions in any animal on the study. The most significant histopathologic lesion, hepatocellular pleomorphism, was observed primarily in the high dose female and, to a lesser extent, male rats. The received study results were recalculated to EDA and then calculated for trimethylenediamine (read across approach).

The resulting effect levels are a NOAEL of 27.9 and a LOAEL of 144 mg/kg bw/day trimethylenediamine.

The read across approach is justified as trimethylenediamine is structurally very similar to ethylenediamine (difference: chain length / one CH2-group) or ethylenediammonium dichloride (applied doses of ammonium and chloride of no toxicological relevance), respectively.

The received effects levels as supported by two other results (also after read across) from studies performed with ethylenediamine (Peters, 1982: 90 days gavage study. NOAEL could not be established. LOAEL 123 mg/kg bw/day trimethylenediamine; effect: eye lesions) and with ethylenediammonium dichloride (Hermansky, 1999: 2 years feeding study. NOAEL: 11 mg/kg bw/day trimethylenediamine. LOAEL 55.7 mg/kg bw/day trimethylenediamine. Effects: Decreased body weight gain and organ weights, hepatocellular pleomorphism, rhinitis, tracheitis).

Repeated dose toxicity: inhalation

In a 6 week study four exposure groups of 30 Sherman rats (15 female/15 male) were exposed to EDA vapour (484 ppm, 225 ppm, 182 ppm and 59 ppm) 7 h /day for 5 days per week. Each exposure group had a control group (Pozzani, 1954). Based on the result for EDA (analytical), the NOAEC was calculated to be 177.6 mg/m³ air trimethylenediamine (read across).

Repeated dose toxicity: dermal

The doses of EDA were selected in preliminary 2-week studies in which various concentrations, 1 to 10%, were applied daily. The skin was closely observed for signs of irritation, and the mice were weighed several times to assess any effects on weight gain. Application of a 5% solution resulted in open sores on the skin of 80% of the treated mice. The 1% solution was the highest EDA concentration which resulted in neither gross skin irritation nor reduced weight gain and was, therefore, chosen for the lifetime dermal carcinogenicity study in 50 male mice (application of 25 µL three times a week). Two EDA samples (purity 99.91% and 99.1%) from different producers were tested. There were no treatment-related macroscopic or histopathologic findings (DePass, 1984). Based on the result for EDA, the dermal chronic NOAEL was calculated to be 10.2 mg/kg bw/day trimethylenediamine.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Most reliable read across study.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
A reliable read across study

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
A reliable read across study.

Repeated dose toxicity: via oral route - systemic effects (target organ) digestive: liver

Repeated dose toxicity: dermal - systemic effects (target organ) other: skin

Justification for classification or non-classification

Based on the available data no classification for repeated dose toxicity / organ toxicity is warranted according to Directive 67/548/EEC (DSD) and Regulation (EC) No 1272/2008 (CLP, GHS).