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Toxicological information

Toxicity to reproduction

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Administrative data

two-generation reproductive toxicity
based on test type (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Very well documented publication. As part of the EDA toxicology program, the present study was undertaken to evaluate the potential reproductive toxicity of EDA in Fischer 344 rats exposed to dietary EDA • 2HG for two generations.

Data source

Reference Type:

Materials and methods

Test guideline
equivalent or similar to
EU Method B.35 (Two-Generation Reproduction Toxicity Test)
study performed in USA, with silmilar set up, very well documented
GLP compliance:

Test material

Details on test material:
Approximately 80 kg of ethylenediamine dihydrochloride (EDA.2HCl) was received from Union Carbide Corporation (UCC, Chemicals and Plastics Division, South Charleston, West Virginia.

Test animals

Fischer 344
Details on test animals and environmental conditions:
Three hundred twenty Fischer 344 rats (170 males, 150 females) were purchased from the Charles River Breeding Laboratories, Inc., Portage, Michiga n. All rats were identified by a toe-clip method and were housed in suspended wire-bottom and wire-front stainless steel cages. Two or three males or three or five females were housed per cage prior to mating; food and water were available ad libitum. Municipal tap water was provided
by an automatic dispensing system with demandcontrol valves in each cage. Ground diet was supplied in 12-oz opal glass jars. The animals were maintained in a room that was kept at approximately 22°C and 55% relative humidity and on a 12-hr fluorescent light cycle.
The Fo rats were 43 day* of age at first exposure. Bodyweight ranges for the males and females at the start of the study were 112-141 and 85-111 g, respectively.

Administration / exposure

Route of administration:
oral: feed
other: ground Purina Certified Rodent Chow 5002
Details on exposure:
For the Fo generation, 25 male rats and 26 female rats were fed a diet containing EDA • 2HC1 at one of three dosage levels (0.50,0.15, or 0.05 g/kg/day) while one group of 50 males and 52 females was fed a control diet.
For the F1 generation, 15 male rats and 26 female rats were fed a diet containing EDA • 2HC1 at one of three dosage levels (0.50, 0.15, or 0.05 g/kg/day) while one group of 30 males and 52 females was fed a control diet.

The basic diet was ground Purina Certified Rodent Chow 5002. Fresh diet was prepared biweekly with the percentage of EDA • 2HQ in the diet adjusted to maintain a constant dosage level in grams per kilogram for each sex according to the average animal body we ight gain and diet consumption.
Since earlier unpublished results from this laboratory indicated that the diet consumption of the dam during Lactation Weeks 2 and 3 increased approximately two- and threefold respectively, the concentration of EDA • 2HC1 in the diet during Lactation Weeks 2 and 3 were adjusted downward two- and threefold accordingly to avoid excessive dosing of the dam.

Details on mating procedure:
The first generation, designated Fo, served as parents for the F, offspring which subsequently became the parents of the next generation. The study was terminated when the offspring of the last mating trial (FJ were weaned and sacrificed.
Duration of treatment / exposure:
The Fo parents received their first exposure at 43 days of age. Diet consumption was determined on each cage of animals every 2 weeks, while the body weight of each animal on the study was measured weekly. The Fo parents were mated after having received dietary EDA • 2HC1 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 mate; at this time all rats received EDA • 2HC1 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. Ab-sorb-dri hardwood chips were used for bedding in the shoebox cages. Diet consumption was determined on each lactating female during Lactation Weeks 1, 2, and 3. The Fo males were used in a related EDA • 2HC1 dominant lethal study (Slesinski et at.. 1983) following the successful establishment of the F, generation.
Details on study schedule:
The Fo rats were assigned to treatment groups using a computer-generated random number scheme. Treatment effects were determined by statistical comparison of mo rtality, body weight gain, food consumption, organ weight change, and histologies] evaluation of tissues from sacrificed animals. In addition, the following reproductive parameters were evaluated statistically, fertility index, days from first mating to parturition, gestation index (the fraction of pregnancies that resulted in litters with live pups), gestation survival index (the fraction of newborn pups that were alive at birth), 0- to 4-day survival index (viability index), 4- to 14-day survival index, 4- to 21-day survival index (lactation index), pups bom alive per litter, pup body weight (by litter) at Lactation Day 4, pup body weight (by litter) at Lactation Day 14, and individual pup body weight at Weaning Day 21
Doses / concentrations
Doses / Concentrations:
0.50, 0.15, 0.05, 0 g/kg/day
nominal in diet

Results and discussion

Results: F2 generation

Effect levels (F2)

Dose descriptor:
Effect level:
ca. 150 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: No indication of reproductive effects, (mating after 100 days of exposure). NOAEL = 150 mg/kg/day

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

No indication of reproductive effects, (mating after 100 days of exposure).
NOAEL = 150 mg/kg/day
Executive summary:

Ethylenediamine dihydrochloride (EDA • 2HC1) was incorporated in the diet of Fischer 344 rats at dosage goals of 0.50, 0.15, 0.05, or 0 g/kg/day in a two-generation reproduction study. The parent generation (Fo) and the F1 generation were each bred once.

Parameters examined included indices of fertility, gestation of dams, gestation survival, 0- to 4-, 4- to 14- and 4- to 21-day survival of pups, number of pups born alive, and number of pups weaned per litter. Furthermore, observations were made on mortality, diet consumption, and body weight of the adult rats in Fo and F, generation. Randomly selected F, weanlings and adults and F2 weanlings were sacrificed and organ weights were obtained; in addition, gross and histologic examinations were conducted on these rats. No reproductive toxicity was observed in this study. Some effects were observed in both sexes for the Fo and F1 parent rats. These effects were mainly associated with the high dosage level and included reduction of body weight gain and changes in liver (decrease) and kidney (increase) weights in the adult rats. The only microscopic lesion observed was hepatocellular pleomorphism in the high level F, adult males and females; a greater prevalence and severity of this lesion was seen in the female rats.

NOAEL = 150 mg/kg/day (mating after 100 days of exposure)