Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
two-generation reproductive toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
before 1984
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
Unnamed
Year:
1984

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)
GLP compliance:
not specified
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Ethylenediamine
EC Number:
203-468-6
EC Name:
Ethylenediamine
Cas Number:
107-15-3
Molecular formula:
C2H8N2
IUPAC Name:
ethane-1,2-diamine
Details on test material:
- Name of test material (as cited in study report): Ethylendiamine
- Analytical purity: 99 %
Specific details on test material used for the study:
Due to corrosivity of EDA, ethylene diammonium dichloride (ECDA.2HCl) was used (CAS no. 333-18-6; EC no. 206-369-6).

Test animals

Species:
rat
Strain:
Fischer 344
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Source: Charles River Breeding Laboratories, Inc., Portage, Michigan.
- Age at study initiation: (P) 43 wks;
- Weight at study initiation: (P) Males: 112-141 g; Females: 85-111 g;
- Housing: Two or three males or three or five females were housed per cage prior to mating;
- Diet: ad libitum, Purina Certified Rodent Chow 5002.
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 55
- Photoperiod (hrs dark / hrs light): 12 hr light cycle

Administration / exposure

Route of administration:
oral: feed
Details on exposure:
Fresh diet was prepared biweekly with the percentage of EDA*2HCl in the diet adjusted to maintain a constant dosage level in grams per kilogram for each sex according to the average animal body weight gain and diet consumption. At monthly intervals two diet samples were analysed.
Details on mating procedure:
The F0 parents were mated afier having received dietary EDA * 2HCl for 100 days. Twenty-six females and 13 males (randomly selected from 25 males) were mated per treated group while 52 females and 26 males (randomly selected from 50 males) were mated for the control group. During mating, two females were placed in a cage with one male; at this time all rats received EDA * 2HCl at the concentration in the diet for the respective female groups. Fifteen days after cohabitation, mating was discontinued and the females were placed in individual shoebox cages fitted with wire rod metal tops.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
At monthly intervals two diet samples were analysed.
Duration of treatment / exposure:
for two generations
Frequency of treatment:
daily
Details on study schedule:
At each dose level 13 male and 26 female rats were mated in both F0 and F1 generation (control group: 26 male and 52 female rats each); continuous treatment starting 100 days prior to cohabitation of F0 rats until weaning of F2 rats.
Doses / concentrationsopen allclose all
Dose / conc.:
50 mg/kg bw/day
Dose / conc.:
150 mg/kg bw/day
Dose / conc.:
500 mg/kg bw/day
No. of animals per sex per dose:
13 male and 26 female. Control group: 26 male and 52 female rats
Control animals:
yes, concurrent no treatment

Examinations

Parental animals: Observations and examinations:
BODY WEIGHT: Yes, measured weekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Food consumption determined on each cage of animals every 2 week. Dosage adjusted to keep a constant dosage level.

Parameters examined included indices of fertility, days from first mating to parturition, gestation index.
Litter observations:
Parameters examined included survival rate on lactation day 4, 14 and 21, pups born alive/litter, pup body weight (by litter) at lactation day 14 and individual pup body weight at weaning day 21.
Postmortem examinations (parental animals):
Complete necropsies were performed on 10 adults/sex/ dose group of F1 generation and on 20 rats/sex of control group. parameters examined included indices of fertility, days from first mating to parturition, gestation index, survival rate on lactation day 4, 14 and 21, pups born alive/litter, pup body weight (by litter) at lactation day 14 and individual pup body weight at weaning day 21.
Postmortem examinations (offspring):
Complete necropsies were performed on 5 weanlings from both F1 and F2 generation at each dose group.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Mortality:
no mortality observed
Description (incidence):
No substance-related parental deaths in the F0 or F1 generation.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Significant reduction in parental body weight gain of female rats in the intermediate and high dose group of the F0 generation, in the high dose group of the F1 generation and of male rats in the high dose group of both F0 and F1 generations.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
No histopathologic findings except for a significant higher incidence of hepatocellular pleomorphism in both sexes of the high dose group of the F1 generation (6/10 male, 10/10 female; control: 0/20 each) and a significant decreased prevalence of kidney tubular mineralization in female rats of the high dose group of the F1 generation (0/10 female; control: 10/20).
Other effects:
no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
23 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Changes in body weight, food consumption, organ weights, and histopathology at the next higher dose (recalculated to EDA base)
Dose descriptor:
NOAEL
Effect level:
>= 227 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects on reproductive performance (recalculated to EDA base)

Results: P1 (second parental generation)

Effect levels (P1)

open allclose all
Dose descriptor:
NOAEL
Effect level:
23 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Changes in body weight, food consumption, organ weights and histopathology at the next higher dose (recalculated to EDA base)
Dose descriptor:
NOAEL
Effect level:
>= 227 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects on reproductive performance (recalculated to EDA base)

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
>= 227 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: absence of embryotoxicity (recalculated to EDA base)

Results: F2 generation

Effect levels (F2)

Dose descriptor:
NOAEL
Generation:
F2
Effect level:
>= 227 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: absence of embryotoxicity (recalculated to EDA base)

Overall reproductive toxicity

Reproductive effects observed:
not specified

Any other information on results incl. tables

Significant reduction in parental body weight gain of female rats in the intermediate and high dose group of the F0 generation, in the high dose group of the F1 generation and of male rats in the high dose group of both F0 and F1 generations.

No substance-related parental deaths in the F0 or F1 generation.

Significant decrease of absolute liver weight in male rats of the high dose F1 generation.

No macroscopic or histopathologic findings except a significant higher incidence of hepatocellular pleomorphism in both sexes of the high dose group of the F1 generation (6/10 male, 10/10 female; control: 0/20 each) and a significant decreased prevalence of kidney tubular mineralization in female rats of the high dose group of the F1 generation (0/10 female; control: 10/20).

In conclusion there was no evidence of fertility impairment or embryotoxic effect at dose levels that show maternal or paternal toxicity.

REPRODUCTIVE INDICES        
         
  Dosage (g/kg,lday)
  0.50 0.15 0.05 0
  F0 → F1
Fertility index (%)        
Male 92 100 92 92
Female 81 77 73 77
Gestation index (%)  100 100 100 100
Gestation survival index (%)  99.0 ± 3.4  100.0 ± 0.0  97.2 ± 11.8  99.2 ± 5.2
0- to 4-Day survival index (%)  90.0 ± 30.0  100.0 ± 0.0  94.4 ± 23.6  93.2 ± 22.4
4- to 14-day survival indexb (%)  99.4 ± 2.5  100.0 ± 0.0  94.1 ± 24.3  100.0 ± 0.0
4- to 2I-day survival indexb (%)  99.4 ± 2.5  100.0 ± 0.0  94.1 ± 24.3  99.7 ± 2.0
Pups born alive/litter  6.8 ± 3.5  8.6 ± 2.8  6.2 ± 3.9  8.4 ± 3.0
Days from first mating to parturition 28.3 ± 3.8 29.6 ± 2.0  29.8 ± 3.0  28.1 ± 3.0
  F1 → F2
Fertility index (%)        
Male  100 92 92 100
Female  92 88 73 88
Gestation index (%)  100 100 100 100
Gestation survival index (%)  98.6 ± 6.7  99.1 ± 3.1  99.4 ± 2.5  99.5 ± 2.0
0- to 4-Day survival index (%) 98.0 ± 4.1  99.3 ± 3.5 98.6 ± 3.3  98.5 ± 4.1
4- to 14-day survival index (%) 100.0 ± 0.0  98.7 ± 6.3 100.0 ± 0.0  99.6 ± 3.0
4- to 21-day survival index (%) 100.0 ± 0.0  98.3 ± 8.3 100.0 ± 0.0  99.6 ± 3.0
Pups born alive/litter 8.5 ± 3.0  8.8 ± 2.2 9.4 ± 2.8 10.0 ± 2.5
Days from first mating to parturition 28.5 ± 3.2  28.8 ± 3.3 27.3 ± 3.6  28.4 ± 3.8

Applicant's summary and conclusion

Conclusions:
There was no indication of reproductive and embryotoxicity in Fischer 344 rats, following exposure to dietary EDA for two generations.
Executive summary:

In a two-generation study, male and female Fischer 344 rats were fed diets containing 0, 50, 150 or 500 mg/kg bw/day of EDA*2HCl. At each dose level 13 male and 26 female rats were mated in both F0 and F1 generation. In the high dose group effects noted were decreased body weight gain in both F0 and F1 generations, a higher incidence of hepatocellular pleomorphism in F1 generation, and decreased liver weights in male rats in F1 generation. In the intermediate dose group female rats of F0 generation showed a decreased body weight gain. No evidence of impaired fertility or embryotoxicity was seen at any dose level. NOAEL parental toxicity was 50 mg/kg bw/day (23 mg/kg bw/day calculated as Ethylenediamine base), NOAEL reproductive performance at least 500 mg/kg bw/day (227 mg/kg bw/day calculated as Ethylenediamine base) and NOAEL F1 offspring was at least 500 mg/kg (227 mg/kg bw/day calculated as Ethylenediamine base; highest dose tested).