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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not applicable
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline and GLP study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report date:
1993

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: OECD draft guideline no. 421
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
2,2'-iminodi(ethylamine)
EC Number:
203-865-4
EC Name:
2,2'-iminodi(ethylamine)
Cas Number:
111-40-0
Molecular formula:
C4H13N3
IUPAC Name:
N-(2-aminoethyl)ethane-1,2-diamine
Details on test material:
- Physical state: colorless to light straw
DETA 99.4%
pH: 12.5 (25% solution)
Batch number: 0511658A

Test animals

Species:
rat
Strain:
Wistar
Details on test animals or test system and environmental conditions:
Ninety-one virgin female, and 95 male Wistar W.U. albino rats of about 6 weeks old, bred under SPF-conditions, were obtained from Charles River Wiga, Sulzfeld, Germany. Upon arrival, the animals were taken in their unopened shipping boxes to a quarantine room, and checked for overt signs of ill health and anomalies. Also a serological control was conducted. Once the results of the serology tests were known and satisfactory, the animals were first moved to study room and then subsequently to a study room. In this room they were acclimatized to the animal room conditions for a period of 14 days (preliminary study) or 30 days (reproduction study) .

HOUSING
Upon arrival in the study room, males and females were housed in groups of four per sex per group in suspended stainless steel cages (45x32~18c m) with wire mesh floor and front.
Preliminary study:
The animals assigned to the preliminary study were housed in groups of four per sex per group in similar stainless steel cages (45x32x18 cm).

Reproduction study:
During the premating period of the reproduction study, the animals were housed individually in similar but smaller cages (18x32x18 cm). During mating the animals were housed in pairs of either sex belonging to the same dose group in these cages (18x32x18 cm). Upon evidence of mating, males and females were again housed individually. Around day 18 of gestation the females were moved to macrolon cages (45x32x18 cm) with a wire-mesh top and were provided with saw-dust for the remainder of the gestation period and for the lactation period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5 and 23.5
- Humidity (%): 55 and 70%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: capsule
Vehicle:
other: micropore filtered water
Details on exposure:
Diethylene triamine was administered daily by gavage to male and female Wistar rats at levels of 30, 100 and 300 mg/kg b.w. during a 2-week premating period, and during mating and gestation up to day 4 post partum or at least during a 4-week period, to screen its effect on male and female reproductive performance and on the development of the offspring .
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Analysis for the stability of the test substance in the vehicle showed that DETA was stable in the vehicle for at least 10 days when stored at approx. 4ºC. The content of the samples taken from the solutions actually given to the animals showed that generally the concentrations of DETA were close to the nominal values.
Details on mating procedure:
Premating exposure period (male and female): 2 weeks

After a 2 week premating period, the males and females were paired 1:1 within the same dose groups. Vaginal smears were taken the next morning to determine whether mating had occurred. If no s erm cells were detected, the female in question was placed again with the male. Upon the presence of sperm in the smear or the finding of a vaginal plug, the females were housed individually, and this moment was considered as day 0 of gestation. This procedure was continued until the females were sperm-positive, or 2 weeks had elapsed.
Duration of treatment / exposure:
29-54 days
Frequency of treatment:
Once per day. Daily by gavage to male and female Wistar rats during a 2-week premating period, and during mating and gestation up t o day 4 post partum or at least during a 4-week period,
Duration of test:
2-4 weeks
No. of animals per sex per dose:
The preliminary study was carried out with 20 male and 20 female rats.

The reproduction study was carried out with 48 male and 48 female rats.
Control animals:
yes
Details on study design:
The preliminary study was carried out with 20 male and 20 female rats, selected from a group of 30 males and 30 females. At the start of the study the body weights of the males ranged from 271.3 to 335.0 g, and of the females from 180.5 to 218.8 g. The reproduction study was carried out with 48 male and 48 female rats, selected from a group of 56 males and 56 females. At the start of the study the body weights of the males ranged from 296.2 to 362.3 g, and of the females from 192.5 to 228.8 g.

At the end of the study, the non-pregnant females were sacrificed well after the date of expected delivery, and the pregnant females when their pups were 4 days old

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: at least once a day


BODY WEIGHT: Yes
- Time schedule for examinations:daily during the study, just prior to dosing, and at the end of the study at autopsy


FOOD CONSUMPTION: Yes
- Food consumption of mated females was determined for the following periods: gestation day 0-7, 7-14, and 14-21, and lactation day 1-4.


POST-MORTEM EXAMINATIONS: Yes


LITTER OBSERVATIONS
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:

- Number of implantations: Yes
Fetal examinations:
- Pup weight
- Pup observations
Statistics:
Clinical findings of the adult rats were evaluated by Fisher's exact probability test.
Body weights and food consumption were subjected to one-way analysis of variance (ANOVA) followed by Dunnett's Multiple Comparison test.
Fisher's exact probability test was used to evaluate the numbers of: mated and pregnant females, females with liveborn pups, females surviving delivery, females with stillborn pups or lost litters, liveborn and stillborn pups, pups lost at various stages, pups surviving 4 days, and male pups on postpartum days 1 and 4.
Indices:
mating, fertility, fecundity, gestation, live birth, viability, sex ration day
Historical control data:
No data

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
Mean parental body weights of females in t h e 300 mg/kg b.w. DETA group were slightly decreased when compared to the control values. For the females of this high dose group body weight change during the premating period (days 0-14) and during gestation (days 0-21) showed a statistically significantly decrease when compared to the control females in both periods.

Food consumption data showed a statistically significant decrease in the females of the 300 mg/kg b.w. DETA group.

Maternal performance of the females in the 100 mg and 300 mg/kg b.w. DETA groups were affected: duration of gestation was increased statistically significantly, and post-implantation loss was increased in a dose-related way.

Only one female, in the 300 mg DETA group (D89), had a stillborn pup.

post-implantation loss for the 100 mg and 300 mg DETA groups was increased. For the 300 mg DETA group the increase was statistically significant, and not within the historical range. The sex ratio was slightly high, but similar in all groups.

Effect levels (maternal animals)

Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No adverse effects were observed concerning the general health and the body weights of the pups.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
30 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: Developmental toxicity

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Any other information on results incl. tables

Data obtained through clinical observations, and macroscopic and microscopic examinations, revealed no effects of the treatment.

Applicant's summary and conclusion

Conclusions:
On the basis of the results obtained it can be concluded that:
- Diethylenetriamine (DETA), when administered by gavage for 4 weeks, causes a light parental toxicity at levels of 300 mg/kg b.w. Consequently, the no-adverse effect level for parental toxicity as established in this study is 100 mg/kg b.w. per day.
- In view of the reproduction effects, the no-adverse effect level for reproduction and development is 30 mg/kg b.w. per day.
Executive summary:

Diethylenetriamine (DETA) was administered by gavage to male and female Wistar rats at levels of 0, 100, 300, 500 and 1000 mg/kg b.w. during 7 days to set dose levels for a reproduction/developmental toxicity screening study (OECD draft guideline no. 421). Animals in the 1000 mg DETA group showed a severe reaction to the treatment, as indicated by the mortality rate and macroscopic observations. Animals in the 500 mg DETA group showed a moderate reaction as concluded from pathological examination and decreased body weight. For the reproduction/developmental study levels of 30, 100 and 300 mg/kg b.w./day were chosen.

Diethylenetriamine was administered daily by gavage to male and female Wistar rats at levels of 30, 100 and 300

mg/kg b.w. during a 2-week premating period, and during mating and gestation up to day 4 post partum or at least

during a 4-week period, to screen its effect on male and female reproductive performance and on the development of

the offspring.

During the reproduction study no mortality occurred that could be ascribed to the administration of DETA.

Data obtained through clinical observations, and macroscopic and microscopic examinations, revealed no effects of the treatment.

Mean parental body weights of both males and females in the 300 mg/kg b.w. DETA group were slightly decreased when compared to the control values. Body weight change of the males of the 300 mg DETA group was statistically significantly decreased when compared to the males from the control group from days 0-28. For the females of this high dose group body weight change during the premating period (days 0-14) and during gestation (days 0-21) showed a statistically significantly decrease when compared to the control females in both periods.

Food consumption data showed a statistically significant decrease in the females of the 300 mg/kg b.w. DETA group.

Maternal performance of the females in the 100 mg and 300 mg/kg b.w. DETA groups were affected: duration of

gestation was increased statistically significantly, and post-implantation loss was increased in a dose-related way.

No adverse effects were observed concerning the general health and the body weights of the pups.

On the basis of the results obtained it can be concluded that:

- Diethylenetriamine (DETA), when administered by gavage for 4 weeks, causes a light parental toxicity at levels of 300 mg/kg b.w. Consequently, the no-adverse effect level for parental toxicity as established in this study is 100 mg/kg b.w. per day.

- In view of the reproduction effects, the no-adverse effect level for reproduction and development is 30 mg/kg

b.w. per day.