Tripropylamine

Toxicity to reproduction

Administrative data

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:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study; no data regarding GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

yes

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OPPTS 870-3650

Principles of method if other than guideline:

No data on detailed clinical observation, sensory reactivity to stimuli, assessment of grip strength, and motor activity

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Trimethylamine

EC Number:

200-875-0

EC Name:

Trimethylamine

Cas Number:

75-50-3

Molecular formula:

C3H9N

IUPAC Name:

N,N-dimethylmethanamine

Details on test material:

CAS 75-50-3 (trimethylamine), 30.8% trimethylamine solution, SOURCE: Mitsubishi Gas Chemical Company, Inc. (Niigata; lot # M381012)
The solution contained < 10 ppm of dimethylamine as impurities. Stable for at least 8 days under refrigerated and shielded conditions with a concentration of between 0.08 and 10w/v%.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

-Age: 9 week old

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

no details given

Details on mating procedure:

- M/F ratio per cage: 2
- Length of cohabitation: 2 weeks
- Proof of pregnancy: [vaginal plug and sperm in vaginal smear]

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

42 days; Females: 2 weeks prior to breeding, continuing through breeding (2 weeks), gestation (3 weeks), lactation (4 days), and until the day of necropsy (test day 40 or 54).

Frequency of treatment:

Once daily

Details on study schedule:

Not applicable

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 8, 40, 200 mg/kg/day (in water)
Basis:
no data

No. of animals per sex per dose:

13/sex/dose group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
- Rationale for animal assignment (if not random):
- Other:

Positive control:

no

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: General condition was observed at least once a day during breeding and at least twice a day before and after dosing over the administration period.


DETAILED CLINICAL OBSERVATIONS: No data


BODY WEIGHT: Yes
- Time schedule for examinations: Body weights for males were determined on days 1, 7, 14, 21, 28, 35, 42 and on the day of necropsy. Body weights were determined for all females on days 1, 7, 14. Females who took time before mating were weighed on days 35 and 42. Females who copulated were weighed on pregnancy days 0, 7, 14 and 20. Females who delivered were weighed on nursing days 0, 4, and on the day of necropsy. Females who copulated but did not deliver were weighed on the equivalent of pregnancy day 25 (day of necropsy).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):

Food consumption was measured on days 1, 7, 13, 29, 35 and 41 for males, and on days 1, 7, and 13 for all females. Females with unconfirmed copulation were measured for food consumption on days 29, 35 and 41.

OTHER:
- Urinalysis was conducted on 5 rats/sex/dose level at week 6.
- Hematology and clinical chemistry: Males prior to necropsy and on the  following day after day 42 administration; Females prior to necropsy:  females who delivered-following nursing day 4, females who mated but did  not deliver-equivalent of pregnancy day 25, and females who did not mate-  following day 54 of administration

Oestrous cyclicity (parental animals):

 Estrous cycle was observed from the vaginal smear and  the mean number of days of mating season was calculated..

Sperm parameters (parental animals):

no data

Litter observations:

-Number of delivered pups:  The number of delivered pups (live pups +  stillborns) was counted on nursing day 0, and the delivery rate ((number  of delivered pups/number of implantation traces) x 100) and the live  birth rate ((number of delivered live pups/number of implantation traces)  x 100) were calculated. With the delivered pups, external malformation  and the sex were checked and the sex ratio ((number of male live  pups/number of female live pups) x 100) was calculated.
-Number of dead pups: The number of dead pups was checked daily, and the  birth rate ((number of delivered live pups/number of delivered pups) x  100) and the viable rate of new-born pups on day 4 ((number of live pups  on nursing day 4/number of live pups on nursing day 0) x 100) were  calculated. Dead pups were autopsied and abnormality and internal organs  were observed

Postmortem examinations (parental animals):

ORGANS EXAMINED AT NECROPSY (MACROSCOPIC AND MICROSCOPIC):
-Macroscopic: organ weights: testes and epididymides; females, ovaries and uteri were extracted, the pregnancy corpora lutea number of the ovary was counted under the stereoscopic microscope, the implantation number of the uterus was counted, and the implantation rate ((implantation number/pregnancy corpora lutea number) x 100) was calculated.
-Microscopic: 5 animals/sex/control and high dose group- sperm and prostrate ventral lobes of all males and vagina, ovaries and uteri of all females; also testes, epididymides, ovaries found to be abnormal during pathologic examinations were all examined histopathologically.

Postmortem examinations (offspring):

no details given

Statistics:

Fisher's Exact Test- mating and conception rate,  Mann-Whitney U Test (2-tailed) and Fisher's Exact Test (1-tailed)-  histopathological examinations, Dunnett's Multiple Comparison Test  (significance level=5%)- body weight, food consumption, hematology,  clinical chemistry and organ weights

Reproductive indices:

-Copulation: When mating was confirmed that day was reckoned as pregnancy  day 0.  From the results of copulation, mating rate ((number of animals  mated/number of animals cohabited) x 100), conception rate ((number of  animals conceived/number of animals mated) x 100), number of days needed  for mating from the start of cohabitation and the number of times of  estrus which recurred during that time were calculated.

-Observation of delivery: With all cases where delivery was confirmed,  the pregnancy period (number of days from pregnancy day 0 to delivery  day) was calculated and the birth rate ((number of females who delivered  live pups/number of animals conceived) x 100) for each group was found. 

Offspring viability indices:

-Number of delivered pups:  The number of delivered pups (live pups +  stillborns) was counted on nursing day 0, and the delivery rate ((number  of delivered pups/number of implantation traces) x 100) and the live  birth rate ((number of delivered live pups/number of implantation traces)  x 100) were calculated. With the delivered pups, external malformation  and the sex were checked and the sex ratio ((number of male live  pups/number of female live pups) x 100) was calculated.
-Number of dead pups: The number of dead pups was checked daily, and the  birth rate ((number of delivered live pups/number of delivered pups) x  100) and the viable rate of new-born pups on day 4 ((number of live pups  on nursing day 4/number of live pups on nursing day 0) x 100) were  calculated. Dead pups were autopsied and abnormality and internal organs  were observed

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Details on results (P0)

-Mortality and time to death: 1 male given 200 mg/kg/day died on day 25, 1 male given 200 mg/kg/day died on day 42 and 1 female died on pregnancy day 22 (administration day 38)
-Clinical signs prior to death (200 mg/kg/day): Day 25 male, showed salivation, emaciation, abnormal breathing noise and dyspnea from administration day 19, and vulval periphery fur soil and loose passage were observed from the day before the death. Day 42 male, showed salivation, emaciation, abnormal breathing noise, dyspnea, a drop in body temperature, faded auricle, tottering and brown soil around the nose were observed from administration day 10, although discontinuously. The female was observed with salivation and abnormal breathing noise sporadically from administration day 11.
-Clinical signs in surviving animals: 200 mg/kg/day: Males- salivation 10/13, abnormal breathing noise 3/13, and decreased contact response 1/13; Females-salivation 10/13, abnormal breathing noise 3/13, and emaciation 1/13. 40 mg/kg/day: no abnormalities
-Body weights: Males- No significant differences in body weights at 200 mg/kg/day; however, body weight gains were decreased when compared to controls. There were no significant differences in body weight or body weight gains in males administered 40 mg/kg/day. Females- No significant differences in body weight or body weight gains.
There was no effect of trimethylamine administration on body weights and food consumption of the females and on organ weights, urine examination and hematological examination results in the males and females
-Clinical chemistry: Significant increase in urea nitrogen at  40  mg/kg/day

Effect levels (P0)

1

Dose descriptor:

NOAEL

Remarks:

systemic effects

Effect level:

40 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: Systemic toxicity was observed at 200 mg/kg bw

2

Dose descriptor:

NOAEL

Remarks:

fertility

Effect level:

200 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: no effects

Results: F1 generation

General toxicity (F1)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

systemic toxicity at 200 mg/kg bw

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings:

not specified

Details on results (F1)

There were no effects on the viability of the delivered pups, sex ratio, body weight and gross development.

Effect levels (F1)

Dose descriptor:

NOAEL

Remarks:

offspring

Generation:

F1

Effect level:

200 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: no effects on the viability of the delivered pups, sex ratio, body weight and gross development

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

Daily oral administration of trimethylamine by gavage resulted in the death of two males and 1 female administered 200 mg/kg/day. Abnormal breathing noise, salivation immediately after the administration, ulcers and inflammatory changes in the stomach and intestinal tracts, squamous hyperplasia and edema in submucosa were observed in both males and females in the 200 mg/kg/day group. An inhibition tendency in body weight increase, decrease in food consumption, total protein concentration and albumin concentration were also observed in the males in the same group.

There was no effect of trimethylamine administration on body weights and food consumption of the females and on organ weights, urine examination and hematological examination results in the males and females. Therefore it was inferred that the general toxicological NOAEL (No Observed Adverse Effect Level) is 40 mg/kg/day.

Executive summary:

A combined repeated dose toxicity study with the reproduction/developmental toxicity screening test was performed in accordance with OECD guideline 422 (Takashima et al., 2003). Male and female rats (Sprague-Dawley, 13 animals/sex/dose group) were exposed to a daily oral (gavage) administration of trimethylamine at 0, 8, 40, and 200 mg/kg/day during 42 day. Males were exposed for 28 days (2 weeks prior breeding and 2 weeks through breeding). Females were exposed 2 weeks prior breeding, continuing through breeding (2 weeks), gestation (3 weeks), lactation (4 days), and until the day of necropsy (test day 40 or 54).

Each one male given 200 mg/kg/day died on administration day 25, 38, and 42 respectively. Day 25 male, showed salivation, emaciation, abnormal breathing noise and dyspnoea from administration day 19, and vulval periphery fur soil and loose passage were observed from the day before the death. Day 42 male, showed salivation, emaciation, abnormal breathing noise, dyspnoea, a drop in body temperature, faded auricle, tottering and brown soil around the nose were observed from administration day 10, although discontinuously. The female was observed with salivation and abnormal breathing noise sporadically from administration day 11.

Clinical signs in surviving animals: 200 mg/kg/day: Males- salivation 10/13, abnormal breathing noise 3/13, and decreased contact response 1/13; Females-salivation 10/13, abnormal breathing noise 3/13, and emaciation 1/13. 40 mg/kg/day: no abnormalities -Body weights: Males- No significant differences in body weights at 200 mg/kg/day; however, body weight gains were decreased when compared to controls. There were no significant differences in body weight or body weight gains in males administered 40 mg/kg/day. Females- No significant differences in body weight or body weight gains. There was no effect of trimethylamine administration on body weights and food consumption of the females and on organ weights, urine examination and hematological examination results in the males and females.There was a significant increase in urea nitrogen at 40 mg/kg/day.

Maternal toxicity did not occur at 200 mg/kg, and no embryo-/fetotoxicity was observed. No abnormality which can be considered the effects of trimethylamine administration was observed in the reproductive organs, and no effect on the observation results of estrous cycle was found. Also no effect was found on the mating rate, conception rate, the birth rate, pregnancy period, and nursing state, the number of corpora lutea, the number and rate of implantation, the viability of the delivered pups, sex ratio, body weight and gross development. From the above, it was considered that reproductive/developmental toxicity NOEL is 200 mg/kg/day for both males and females and also for delivered pups. The parental systemic toxicity is 40 mg/kg bw.