4-methyl-m-phenylenediamine

Toxicity to reproduction

Administrative data

Endpoint:

fertility, other

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

other information

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: Not fully equivalent or similar to OECD guideline. Not fully reported.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1985

Materials and methods

Test guideline

Qualifier:

no guideline available

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

4-methyl-m-phenylenediamine

EC Number:

202-453-1

EC Name:

4-methyl-m-phenylenediamine

Cas Number:

95-80-7

Molecular formula:

C7H10N2

IUPAC Name:

4-methylbenzene-1,3-diamine

Details on test material:

0, 0.01 and 0.03% 2,4-TDA

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

Male rats (60-70 day old) were randomly distributed into 4 groups of 8-10 rats each and kept at 24ºC on a light-dark cycle of 12 hour. Rats were fed lab chow mixed with 2,4-TDA at concentrations of 0, 0.01 or 0.03%. The animals were housed one per cage and provided with their respective diets and tap water ad libitum. Body weights were measured weekly.

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: Laboratory chow

Details on exposure:

The rats were given diet containing 0, 0.01, or 0.03% 2,4–TDA.

Details on mating procedure:

After 10 weeks of treatment, each male rat was mated with 2 untreated, virgin females. Insemination was confirmed by vaginal smear, and after insemination the females were separately caged. After 1 week the females not inseminated were replaced by 2 new females, and mating continued for another week. A total of 4 females were mated with each male.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data.

Duration of treatment / exposure:

10 weeks, thereafter 4 weeks recovery

Frequency of treatment:

Continuous

Details on study schedule:

See "Further details on study design" for details.

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 0.01, or 0.03% 2,4–TDA.
Basis:
no data

No. of animals per sex per dose:

10 male rats per dosed group, 8 controls

Control animals:

yes, concurrent no treatment

Details on study design:

After 2 weeks conditioning the (60–70)–day old rats were randomly distributed into 4 groups of 8–10 rats. They were kept 1/cage, at 24°C, and with a 12 hr dark/light cycle. The rats were given a proprietary feed containing 0, 0.01, or 0.03% TDA, and water, ad libitum. The 0.03 and 0.01% doses were approximately equal to 5, and 15 mg 2,4–TDA/kg bw/day, respectively. After 10 weeks of treatment, all the animals were returned to the regular proprietary food ration, and each was mated with 2 untreated, virgin females. Insemination was confirmed by vaginal smear, and after insemination the females were separately caged. After 1 week the females not inseminated were replaced by 2 new females, and mating continued for another week. A total of 4 females were mated with each male.

Body weights were measured weekly. The females were killed 14 days after confirmed insemination, and were examined for the presence and number of implantation sites, viable implants, and resorptions. The males used in breeding were sacrificed 2 weeks after mating, ie, 4 weeks after their return to the regular feed. The testes, and epididymides were fixed, stained and examined by light microscopy.

Positive control:

Not used.

Examinations

Parental animals: Observations and examinations:

Not applicable.

Oestrous cyclicity (parental animals):

Not applicable.

Sperm parameters (parental animals):

The testes and epididymides were fixed and analysed for sperm analyses.

Litter observations:

Not applicable.

Postmortem examinations (parental animals):

Not applicable.

Postmortem examinations (offspring):

Not applicable.

Statistics:

The Chi-square technique was used to detect overall differences in the fertility index and mating index. When significant differences were found, pairwise comparisons between control and treatment groups were made by chi-sequare tests (one degree of freedom) using the Yates correction for continuity. The data for viable implants in pregnant females were analysed by analysis of varience. The level of significance was selected as p<0.05.

Reproductive indices:

Not applicable.

Offspring viability indices:

Not applicable.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

Details on results (P0)

Not applicable.

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

Details on results (F1)

Not applicable.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The rats fed 0 and 0.03% 2,4–TDA in the feed over 10 weeks consumed 25 and 20 g feed/day, respectively. After initial acclimatisation the food consumption did not vary significantly over the 10 weeks.

0.03% (5 mg/kg bw):
Retardation in weight gain during weeks 4 -10 probably due to some reduced food consumption (no further details provided). 5/10 males revealed to be infertile, while the remaining animals in this group exhibited generally unimpaired fertility. Mating index significantly decreased: 0.73 (p<0.01), fertility index significantly decreased: 0.41 (p< 0.05). Light-microscopic examinations of the testes revealed focal or diffuse hypospermatocytogenesis in the seminiferous tubules and in cauda epididymides.

0.01% (15 mg/kg bw):
All animals were fertile; mating index: 0.95, fertility index: 0.74

Control: all animals were fertile; mating index: 1.00, fertility index: 0.72

The pregnancies derived from treated males were not significantly different from those produced from control males in terms of viable implants. Also the mean number of resorptions were similar in all of the groups.

Executive summary:

Effects of 2,4-toluenediamine (TDA) on reproduction in adult male Sprague-Dawley rats were evaluated. Diets containing 0, 0.01 and 0.03% TDA were fed ad libitum to experimental animals for 10 week. No signs of toxicity were found. Exposure to the high dose resulted in decreased mating frequency and an increase in infertile matings. Light-microscopic examination of the testes revealed reduced numbers of sperm in the seminiferous tubules and cauda epididymides. These results indicate that TDA is capable of reducing fertility and of exerting an inhibitory or toxic effect on spermatogenesis in the rat.