Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
20. Apr 1999 - 28. Jun 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD 421)
Qualifier:
according to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Boehringer Ingelheim, Pharma KG, Biberach/Riss
- Age at study initiation: 89-90 days
- Weight at study initiation: Males: 399.7 (365.8 - 430.9) g; Females: 262.5 (235.6 - 291.5) g
- Housing: individually in type DK 111 stainless steel wire mesh cages.
- Diet: Kliba Iaboratory diet rat/mouse/hamster ad libitum
- Water: tap water ad libitum
- Acclimation period: 13 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24 °C
- Humidity (%): 30 - 70 %
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): The preparations were prepared twice a week, every 96 hours latest.


Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: maximum 2 weeks
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test substance solutions were analyzed by gas chromatography.
Duration of treatment / exposure:
males: 28 days
females: 46-56 days
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
0, 10, 50, 200 mg/kg bw
Basis:
nominal conc.
No. of animals per sex per dose:
5
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale:
10 mg/kg body weight/day: as the expected no observed adverse effect level.
50 mg/kg body weight/day: as the intermediate dose level.
200 mg/kg body weight/day: as the dose level with possible toxic effects.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily. The nesting, littering, and lactation behavior of the dams was generally evaluated in the mornings in connection with the daily clinical inspection of the dams.


DETAILED CLINICAL OBSERVATIONS: No data


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

Sperm parameters (parental animals):
Parameters examined in P male parental generations:
testis weight, epididymis weight

Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals after themating period.
- Maternal animals: All surviving animals after the last litter of each generation was weaned.


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

Postmortem examinations (offspring):
All surviving pups, all stillborn pups andthose pups that died before schedule, were examined externally, eviscerated and their organs were assessed macroscopically.
Statistics:
DUNNETT-test, FISHER'S EXACT test, WILCOXON-test
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
There were no substance-related mortalities in any of the male and female F0 parental animals in any of the groups. One high dose male animal showed piloerection on study day 2. Another male animal of this test group had respiratory sounds between days 11-23.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
The food consumption of the high dose F0 male animals was statistically significantly reduced during study weeks 0 - 1 and remained slightly reduced for the whole study period without attaining statistical significance. This was in-line with Iowered mean body weight and impaired body weight gain.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
With the exception of two F0 parental females all mated females of test groups 0 - 3 (0, 10, 50, 200 mg/kg body weight/day) became pregnant. Therefore, the fertility index varied between 90 and 100 %. This difference concerning the female fertility index was regarded to be spontaneous in nature and not associated with the treatment of the animals.

ORGAN WEIGHTS (PARENTAL ANIMALS)
The mean absolute and relative weight parameters did not show significant differences when compared with the control group.

GROSS PATHOLOGY (PARENTAL ANIMALS)
A few gross lesions were noted in the forestomach (erosion/ulcer and thickening of wall) in a high dose male; liver (small yellow focus: in a control male, two males of the mid and one of the high dose group; a mid sized necrosis in a mid dose female rat; lungs (atelectasis, diffuse red discoloration or large hematoma in three different males of the high dose group); kidneys (clay colored discoloration or focal unilateral contraction in the cortex of two different high dose males), and spleen (focal grey white deposition in a low dose group male).

Two animals were not pregnant. In one control animal a dilation of the uterine horns may have contributed to this situation.

HISTOPATHOLOGY (PARENTAL ANIMALS)
With the exception of the clay colored discoloration of the kidneys of one animal (male, high dose group), all gross lesions could be correlated with a meaningful microscopic finding. Histopathology of testes, epididymides and ovaries did not reveal any treatment related microscopic finding.
Dose descriptor:
NOAEL
Remarks:
reproductive performance and fertility
Effect level:
200 mg/kg bw/day (nominal)
Sex:
male/female
Dose descriptor:
NOAEL
Remarks:
systemic toxicity
Effect level:
50 mg/kg bw/day (nominal)
Sex:
male
Basis for effect level:
other: body weight; food consumption
VIABILITY (OFFSPRING)
There were no substance-related differences concerning pup viability and mortality.

CLINICAL SIGNS (OFFSPRING)
The F1 pups did not show any clinical signs which could be attributed to the treatment. An unilateral microphthalmia was the only clinical observations which occurred in one high dose pup. However, this finding was considered to be spontaneous in nature in respect to the known historical background data of the rat strain investigated in the present study.

BODY WEIGHT (OFFSPRING)
Mean body weights/body weight gains were not influenced by the test substance administration.

GROSS PATHOLOGY (OFFSPRING)
The macroscopic examination of all pups did not reveal any differences between the test groups neither in the type nor in the number of pup necropsy observations.
OnIy spontaneous findings were seen at necropsy (e.g. post mortem autolysis, dilated renal pelvis and microphthalmia) in a few of the large number of examined F1 pups of all groups including the controls generally without a dose-response relationship.
Dose descriptor:
NOAEL
Remarks:
developmental toxicity
Generation:
F1
Effect level:
200 mg/kg bw/day (nominal)
Sex:
male/female
Reproductive effects observed:
not specified

Under the conditions of this reproduction/developmental toxicity screening test, the oral administration by gavage of 3-Dimethylaminopropyl- amin had no adverse effects on reproductive performance or fertility of the F0 parental animals of all substance-treated groups. There were no signs of developmental toxicity in the progeny of the F0 parents up to and including the high dose group (200 mg/kg body weight/day).

Signs of general, systemic toxicity in the F0 parental animals were confined to the male rats of the 200 mg/kg body weight/day group. Toxicity was characterized by decreased food consumption.

Conclusions:
The NOAEL (no observed adverse effect level) for reproductive performance and fertility was 200 mg/kg bw/day for the F0 parental rats. The NOAEL for general, systemic toxicity of the test substance was 200 mg/kg bw/day for the F0 parental females, while it was 50 mg/kg bw/day for the F0 parental males. The NOAEL for developmental toxicity could be fixed at 200 mg/kg bw/day for the F1 progeny.
Executive summary:

A Reproduction/Development Toxicity Screening Test of  3-Aminopropyldimethylamine (DMAPA) by oral administration to Sprague-Dawley Rats according OECD method 421 was carried out with four groups of 10 male and 10 female rats each. The rats received 0 (control), 10, 50 or 200 mg/kg bw/day; males: 14 days prior to mating and during the 14-day mating period, females: 14 days prior to mating and during the mating period, pregnancy and lactatoin period. Examinations were carried out on mortality, clinical signs, body weight, food consumption, reproduction, macroscopic post mortem findings/organ weights of parent animals and histopathology. Under the conditions of this reproduction/developmental toxicity screening test according OECD method 421, the oral administration of DMAPA by gavage to rats had no adverse effects on reproductive performance or fertility of the F0 parenteral animals of all substance-treated groups. Mating behavior, conception, gestation, parturition and lactation as well as the determined sexual organ weights, gross and histopathological findings of these organs were similar between the substance-treated animals and the corresponding controls. With the exception of each one male and female F0 parental rat in the control group and mid dose group all F0 parental rats proved to be fertile. Thus, the observable difference was regarded to be incidental in nature and not of toxicological or biological concern. Signs of general, systemic toxicity in the F0 parental animals were confined to the male rats of the 200 mg/kg bw/day group. Toxicity was characterized by decreased food consumption. If calculated for the whole treatment period, it was about 6% below the concurrent control value. The high dose F0 parental males showed also impairments in body weight gain during the whole treatment period. If calculated for the entire treatment period (weeks 0 – 4) body weight gain was about 35% lower than the concurrent control value although there was a statistically significant increase in body weight gain during weeks 2 – 3. Indications for a slight impairment in the general state of health were noted in two F0 male animals (piloerection, respiratory sounds). No compound-related adverse effects were observed in female rats up to the highest dose level of 200 mg/kg bw/day. There were no signs of developmental toxicity in the progeny of the F0 parents up to and including the high dose group (200 mg/kg bw/day). Observed differences did not show any biological relevance between the substance-treated groups and the control group. Thus, the NOAEL (no observed adverse effect level) for reproductive performance and fertility was 200 mg/kg bw/day for the F0 parental rats. The NOAEL for general, systemic toxicity of the test substance was 200 mg/kg bw/day for the F0 parental females, while it was 50 mg/kg bw/day for the F0 parental males. The NOAEL for developmental toxicity could be fixed at 200 mg/kg bw/day for the F1 progeny.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Species:
rat
Quality of whole database:
Read across to an analogue substance.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No reproductive toxicity study is available for N, N, N', N'-tetramethyltrimethylenediamine. However, a Reproduction/Development Toxicity Screening Test was performed by oral administration according to the OECD TG 421 with an analogue substance of 3-aminopropyldimethylamine (DMAPA) (BASF AG, 1999).

Sprague-Dawley rats (10/sex/dose) received 0 (control), 10, 50 or 200 mg DMAPA/kg bw/day; males: 14 days prior to mating and during the 14-day mating period, females: 14 days prior to mating and during the mating period, pregnancy and lactation period. Examinations were carried out on mortality, clinical signs, body weight, food consumption, reproduction, macroscopic post mortem findings/organ weights of parent animals and histopathology. Under the conditions the oral administration of DMAPA by gavage to rats had no adverse effects on reproductive performance or fertility of the F0 parenteral animals of all substance-treated groups. Mating behavior, conception, gestation, parturition and lactation as weil as the determined sexual organ weights, gross and histopathological findings of these organs were similar between the substance-treated animals and the corresponding controls. With the exception of each one male and female F0 parental rat in the control group and mid dose group all F0 parental rats proved to be fertile. Thus, the observable difference was regarded to be incidental in nature and not of toxicological or biological concern. Signs of general, systemic toxicity in the F0 parental animals were confined to the male rats of the 200 mg/kg bw/day group. Toxicity was characterized by decreased food consumption. If calculated for the whole treatment period, it was about 6% below the concurrent control value. The high dose F0 parental males showed also impairments in body weight gain during the whole treatment period. If calculated for the entire treatment period (weeks 0 - 4) body weight gain was about 35% lower than the concurrent control value although there was a statistically significant increase in body weight gain during weeks 2 - 3. Indications for a slight impairment in the general state of health were noted in two F0 male animals (piloerection, respiratory sounds). No compound-related adverse effects were observed in female rats up to the highest dose level of 200 mg/kg bw/day. There were no signs of developmental toxicity in the progeny of the F0 parents up to and including the high dose group (200 mg/kg bw/day). Observed differences did not show any biological relevance between the substance-treated groups and the control group. Thus, the NOAEL (no observed adverse effect level) for reproductive performance and fertility was 200 mg/kg bw/day for the F0 parental rats. The NOAEL for general, systemic toxicity of the test substance was 200 mg/kg bw/day for the F0 parental females, while it was 50 mg/kg bw/day for the F0 parental males. The NOAEL for developmental toxicity could be fixed at 200 mg/kg bw/day for the F1 progeny.


Short description of key information:
In the study in rats conducted according to the OECD 421 with the analogue substance, 3-aminopropyldimethylamine, a clear-cut no-observed-adverse-effect level (NOAEL) of 200 mg/kg bw/day for reproductive performance and fertility was determined by gavage

Justification for selection of Effect on fertility via oral route:
Key study

Effects on developmental toxicity

Description of key information
In the study conducted according to the OECD 421 with the analogue substance, 3-aminopropyldimethylamine, a clear-cut no-observed-adverse-effect level (NOAEL) of 200 mg/kg bw/day for developmental toxicity was determined by gavage.
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
20. Apr 1999 - 28. Jun 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
other: OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Boehringer Ingelheim, Pharma KG, Biberach/Riss
- Age at study initiation: 89-90 days
- Weight at study initiation: Males: 399.7 (365.8 - 430.9) g; Females: 262.5 (235.6 - 291.5) g
- Housing: individually in type DK 111 stainless steel wire mesh cages.
- Diet: Kliba Iaboratory diet rat/mouse/hamster ad libitum
- Water: tap water ad libitum
- Acclimation period: 13 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24 °C
- Humidity (%): 30 - 70 %
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The preparations were prepared twice a week, every 96 hours latest.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test substance solutions were analyzed by gas chromatography.
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: maximum 2 weeks
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
Duration of treatment / exposure:
males: 28 days
females: 46-56 days (including gestational day 1 to 21)
Frequency of treatment:
daily
Remarks:
Doses / Concentrations:
10, 50, 200 mg/kg bw
Basis:
nominal conc.
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily. The nesting, littering, and lactation behavior of the dams was generally evaluated in the mornings in connection with the daily clinical inspection of the dams.


DETAILED CLINICAL OBSERVATIONS: No data


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
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: No
Fetal examinations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, weight gain.
All surviving pups, all stillborn pups and those pups that died before schedule, were examined externally, eviscerated and their organs were assessed macroscopically.
Statistics:
DUNNETT-test, FISHER'S EXACT test, WILCOXON-test
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
There were no substance-related mortalities in any of the male and female F0 parental animals in any of the groups. One high dose male animal showed piloerection on study day 2. Another male animal of this test group had respiratory sounds between days 11-23.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
The food consumption of the high dose F0 male animals was statistically significantly reduced during study weeks 0 - 1 and remained slightly reduced for the whole study period without attaining statistical significance. This was in-line with Iowered mean body weight and impaired body weight gain.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
With the exception of two F0 parental females all mated females of test groups 0 - 3 (0, 10, 50, 200 mg/kg body weight/day) became pregnant. Therefore, the fertility index varied between 90 and 100 %. This difference concerning the female fertility index was regarded to be spontaneous in nature and not associated with the treatment of the animals.

ORGAN WEIGHTS (PARENTAL ANIMALS)
The mean absolute and relative weight parameters did not show significant differences when compared with the control group.

GROSS PATHOLOGY (PARENTAL ANIMALS)
A few gross lesions were noted in the forestomach (erosion/ulcer and thickening of wall) in a high dose male; liver (small yellow focus: in a control male, two males of the mid and one of the high dose group; a mid sized necrosis in a mid dose female rat; lungs (atelectasis, diffuse red discoloration or large hematoma in three different males of the high dose group); kidneys (clay colored discoloration or focal unilateral contraction in the cortex of two different high dose males), and spleen (focal grey white deposition in a low dose group male).

Two animals were not pregnant. In one control animal a dilation of the uterine horns may have contributed to this situation.

HISTOPATHOLOGY (PARENTAL ANIMALS)
With the exception of the clay colored discoloration of the kidneys of one animal (male, high dose group), all gross lesions could be correlated with a meaningful microscopic finding. Histopathology of testes, epididymides and ovaries did not reveal any treatment related microscopic finding.
Dose descriptor:
NOAEL
Effect level:
>= 200 mg/kg bw/day (nominal)
Basis for effect level:
other: maternal toxicity
Dose descriptor:
NOAEC
Effect level:
>= 200 mg/kg bw/day (nominal)
Basis for effect level:
other: developmental toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
VIABILITY (OFFSPRING)
There were no substance-related differences concerning pup viability and mortality.

CLINICAL SIGNS (OFFSPRING)
The F1 pups did not show any clinical signs which could be attributed to the treatment. An unilateral microphthalmia was the only clinical observations which occurred in one high dose pup. However, this finding was considered to be spontaneous in nature in respect to the known historical background data of the rat strain investigated in the present study.

BODY WEIGHT (OFFSPRING)
Mean body weights/body weight gains were not influenced by the test substance administration.

GROSS PATHOLOGY (OFFSPRING)
The macroscopic examination of all pups did not reveal any differences between the test groups neither in the type nor in the number of pup necropsy observations.
OnIy spontaneous findings were seen at necropsy (e.g. post mortem autolysis, dilated renal pelvis and microphthalmia) in a few of the large number of examined F1 pups of all groups including the controls generally without a dose-response relationship.
Abnormalities:
not specified
Developmental effects observed:
not specified
Conclusions:
The NOAEL (no observed adverse effect level) for reproductive performance and fertility was 200 mg/kg bw/day for the F0 parental rats. The NOAEL for general, systemic toxicity of the test substance was 200 mg/kg bw/day for the F0 parental females, while it was 50 mg/kg bw/day for the F0 parental males. The NOAEL for developmental toxicity could be fixed at 200 mg/kg bw/day for the F1 progeny.
Executive summary:

A Reproduction/Development Toxicity Screening Test of  3-Aminopropyldimethylamine (DMAPA) by oral administration to Sprague-Dawley Rats according OECD method 421 was carried out with four groups of 10 male and 10 female rats each. The rats received 0 (control), 10, 50 or 200 mg/kg bw/day; males: 14 days prior to mating and during the 14-day mating period, females: 14 days prior to mating and during the mating period, pregnancy and lactatoin period. Examinations were carried out on mortality, clinical signs, body weight, food consumption, reproduction, macroscopic post mortem findings/organ weights of parent animals and histopathology. Under the conditions of this reproduction/developmental toxicity screening test according OECD method 421, the oral administration of DMAPA by gavage to rats had no adverse effects on reproductive performance or fertility of the F0 parenteral animals of all substance-treated groups. Mating behavior, conception, gestation, parturition and lactation as well as the determined sexual organ weights, gross and histopathological findings of these organs were similar between the substance-treated animals and the corresponding controls. With the exception of each one male and female F0 parental rat in the control group and mid dose group all F0 parental rats proved to be fertile. Thus, the observable difference was regarded to be incidental in nature and not of toxicological or biological concern. Signs of general, systemic toxicity in the F0 parental animals were confined to the male rats of the 200 mg/kg bw/day group. Toxicity was characterized by decreased food consumption. If calculated for the whole treatment period, it was about 6% below the concurrent control value. The high dose F0 parental males showed also impairments in body weight gain during the whole treatment period. If calculated for the entire treatment period (weeks 0 – 4) body weight gain was about 35% lower than the concurrent control value although there was a statistically significant increase in body weight gain during weeks 2 – 3. Indications for a slight impairment in the general state of health were noted in two F0 male animals (piloerection, respiratory sounds). No compound-related adverse effects were observed in female rats up to the highest dose level of 200 mg/kg bw/day. There were no signs of developmental toxicity in the progeny of the F0 parents up to and including the high dose group (200 mg/kg bw/day). Observed differences did not show any biological relevance between the substance-treated groups and the control group. Thus, the NOAEL (no observed adverse effect level) for reproductive performance and fertility was 200 mg/kg bw/day for the F0 parental rats. The NOAEL for general, systemic toxicity of the test substance was 200 mg/kg bw/day for the F0 parental females, while it was 50 mg/kg bw/day for the F0 parental males. The NOAEL for developmental toxicity could be fixed at 200 mg/kg bw/day for the F1 progeny.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Species:
rat
Additional information

In the study conducted according to the OECD 421 with the analogue substance, 3-aminopropyldimethylamine, there were no signs of developmental toxicity in the progeny of the F0 parents up to and including the high dose group (200 mg/kg bw/day).

Justification for classification or non-classification

In the study conducted according to the OECD 421 with the analogue substance, 3 -aminopropyldimethylamine, there were no signs of effects on reproductive performance and fertility and on developmental toxicity in the progeny of the F0 parents up to and including the high dose group (200 mg/kg bw/day). Therefore, N, N, N', N'-tetramethyltrimethylenediamine does not warrant classification for reproductive and developmental toxicity according to Regulation (EC) No 1272-2008 and Annex VI of Commission Directive 2001/59/EC.