Amines, C16-18 (even numbered)-alkyl , salts with phosphoric acid, mono- and di-C16-18 (even numbered) alkyl esters

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: EPA regulations, TSCA (40 CFR Part 798.4700, September 1985, and revised edition May 1987)

GLP compliance:

yes

Remarks:

Springborn Laboratories, Inc., Mammalian Toxicology Division, 553 North Broadway, Spencerville, Ohio 45887, USA

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

(Z)-octadec-9-enylamine

EC Number:

204-015-5

EC Name:

(Z)-octadec-9-enylamine

Cas Number:

112-90-3

Molecular formula:

CH3(CH2)7CH=CH(CH2)7CH2NH2

IUPAC Name:

octadec-9-en-1-amine

Constituent 2

Reference substance name:

(Z)-octadec-9-en-1-amine

IUPAC Name:

(Z)-octadec-9-en-1-amine

Constituent 3

Reference substance name:

Oleyl Alkylamines

IUPAC Name:

Oleyl Alkylamines

Details on test material:

- Name of test material (as cited in study report): Oleylamine

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Portage, Michigan
- Age at study initiation: 14 weeks
- Weight at study initiation: 229-316 g
- Housing: Animals were housed individually during acclimatization and while on study, except during cohabitation, in suspended stainless steel wire mesh cages.
- Diet (ad libitum): Purina Certified Rodent Meal #5002
- Water (ad libitum): deionized tap water
- Acclimation period: 19 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): ~22
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12 / 12


IN-LIFE DATES: From: 1989-07-25 To: 1989-08-18

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Mazola® corn oil

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Dose solutions were prepared daily. Appropriate amounts of the test article for each dose group were weighed into volumetric flasks. Mazola® corn oil was added in sufficient quantity to achieve the final concentrations. The flasks were inverted several times to ensure adequate mixture. Dosing solutions were stored under a nitrogen blanket at room temperature.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

On the first day of dosing and near the end of the dosing period, a subsample from each dosing solution, including the control, was taken and analyzed for verification of the test article concentration.

Details on mating procedure:

Female rats determined to be suitable test subjects were cohabitated with proven resident Sprague Dawley Cr1:COBS®CD®BR®VAF/PLUS® male rats. The male rats were of suitable health and were nine to eleven months old. Evidence of mating was determined by the presence of a copulatory plug in the vagina of a sperm positive vaginal smear and was considered day 0 of gestation.

Duration of treatment / exposure:

from gestation day 6 through 15

Frequency of treatment:

single daily doses

Duration of test:

10 days

No. of animals per sex per dose:

28

Control animals:

yes, concurrent vehicle

Details on study design:

Range-finding study: 50, 100, 150, 250 mg/kg/day
Mortality occurred in the 100, 150 and 250 mg/kg/day groups. Outward clinical signs of toxicity and body weight losses or reduced weight gain occurred at the 50, 100, 150 and 250 mg/kg/day levels. A dose level of 100 mg/kg/day was considered to be excessive for a high dose level of the definitive teratology study due to the induced mortality. Conversely, 50 mg/kg/day did not produce sufficient maternal toxicity to be considered suitable as a high dose level. Thus, 80 mg/kg/day was selected in anticipation of producing sufficient maternal toxicity. Graduated doses of 40 and 10 mg/kg/day were selected as the mid and low dose levels, respectively. A dose level of 40 mg/kg/day was expected to induce minimal maternal toxicity while the 10 mg/kg/day dose level was selected to determine a no effect level for maternal and developmental toxicity.

Examinations

Maternal examinations:

Animals were examined daily. Clinical signs of toxicity including physical or behavioral abnormalities were recorded. Mortality checks were performedtwice daily (morning and afternoon). During the dosing period, animals were observed for toxic effects between on-half hour and two hours postdosing.

Body weights were recorded on GD 0, 6, 9, 12, 16 and 20. BWC was recorded for GD 0-6. 6-9, 9-12, 12-16, 16-20, 6-16 and 0-20. Net maternal body weight gain (adjusted for gravid uterine weight) was also reported.

Food consumption was measured on GD 0, 6, 9, 12, 16, 20. Food consumption was calculated as g/kg/day and g/animal/day for the following gestation intervals: 0-6, 6-9, 9-12, 12-16, 16-20, 6-16, 0-20.

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes

Fetal examinations:

Fetal morphological examination: external (each fetus), visceral (one-half of the fetuses from each litter), skeletal (one-half of the fetuses from each litter) abnormalities

Statistics:

Analyses were performed by a digital VAX 11/730 computer. Two-tailed tests were utilized unless otherwise indicated for a minimum significance level of 5% comparing the control group to each treatment group. Group comparisons were performed using Dunnett´s test. Count data were analyzed using Chi-square Test for fetal sex ratios, Mann-Whitney U-Test for resorptions, and Fisher´s Exact Test for the number of fetal variations and malformations.

Historical control data:

Cesarean section data, fetal malformation data, fetal variation data were provided.

Results and discussion

Results: maternal animals

Maternal developmental toxicity

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
Clinical signs of toxicity were observed at the 40 and 80 mg/kg/day levels. Salient clinical signs included salivation. Dose dependent body weight losses or reduced weight gain, along with a corresponding reduction in food consumption occurred.

Effect levels (maternal animals)

1

Dose descriptor:

NOAEL

Effect level:

10 mg/kg bw/day (nominal)

Basis for effect level:

other: maternal toxicity

2

Dose descriptor:

NOAEL

Effect level:

> 80 mg/kg bw/day (nominal)

Basis for effect level:

other: developmental toxicity

Results (fetuses)

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No treatment-related changes were apparent at any level tested concerning necropsy observations, cesarean section data or fetal morphological examinations.

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

> 80 mg/kg bw/day (nominal)

Basis for effect level:

other: teratogenicity

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Any other information on results incl. tables

Gross necropsy evaluation of the gastrointestinal tracts from females sacrificed on gestation day 15 revealed no irritative effects.

The oral administration of 40 and 80 mg/kg/day of Oleylamine to pregnant CD rats, induced dose dependent maternal toxicity exhibited by adverse clinical signs, body weight losses and reduced food consumption. A dosage level of 10 mg/kg/day was determined to be a no effect level for maternal toxicity. Oleylamine was neither developmentally toxic nor teratogenic at dosage levels of 10, 40 and 80 mg/kg/day.

Applicant's summary and conclusion

Conclusions:

Oral administration of 40 and 80 mg/kg body weight per day of Oleylamine to pregnant CD rats induced dose dependent maternal toxicity exhibited by adverse clinical signs, body weight loss and reduced food consumption. A dosage level of 10 mg/kg body weight per day was determined to be a no effect level for maternal toxicity. Oleylamine was neither developmental toxic nor teratogenic at dosage levels of 10, 40 or 80 mg/kg body weight per day.

Executive summary:

For octadecenylamine ("Oleylamine") a guideline conform teratology study in Sprague Dawley rats has been performed. Prior to initiation of the main study, a range-finding study had been conducted. During the range finding-study, treatment-related deaths had occurred in the 100, 150 and 250 mg/kg bw/day groups. Outward clinical signs of toxicity and body weight loss or reduced weight gain occurred at the 50, 100, 150 and 250 mg/kg bw/day dose levels. A dose level of 100 mg/kg bw/day was considered to be excessive for a high dose level of the main study due to the induced mortality. Conversely, 50 mg/kg bw/day did not produce sufficient maternal toxicity to be considered suitable as a high dose level. Thus, 80 mg/kg bw/day was selected for the main study in anticipation of producing sufficient maternal toxicity.

In the main study groups of 28 pregnant females were treated orally (gavage) with dosages of10, 40 or 80 mg/kg bw/day or with the vehicle (Mazola corn oil) during gestation days 6 to 15. During the study animals were examined daily. Any clinical signs of toxicity including physical or behavioural abnormalities were recorded. Individual body weights and food consumption were recorded on gestation days 0, 6, 9, 12, 16, and 20. Two animals in each group were selected to be sacrificed and necropsied after treatment on gestation day 15 to determine the appearance and severity of gastrointestinal tract irritation. On gestation day 20 caesarean section was performed on all surviving animals. The numbers of viable fetuses, early and late resorptions as well as the number of corpora lutea were recorded. Fetuses were examined for external, visceral and skeletal abnormalities.

All animals survived to scheduled sacrifice. Outward clinical signs of toxicity were observed at the 40 and 80 mg/kg bw/d dose levels. The observations most likely indicated a generalised irritative effect of the test substance as characterised by rales, salivation, unkempt appearance and changes in the amount, colour and consistency of the feces. However, no other signs of treatment-related gastrointestinal irritation or other internal changes were observed at the gestation day 15 and 20 necropsies. More pronounced signs of toxicity were apparent only in the 80 mg/kg bw/d dose group and included emaciation, rough coat and dark red material around the eyes, nose and/or mouth. Similar clinical signs were infrequently noted during the post-dose observations. Dose-dependent body weight loss (during gestation days 6-9) or reduced weight gain (during gestation days 12-16), along with a corresponding reduction in food consumption occurred during the treatment period in the 40 and 80 mg/kg bw/d dose groups. Net body weight gain (adjusted for gravid uterine weight) was also lower at these levels. Following cessation of treatment (days 16-20), increase in weight gain and food consumption were noted at both dose levels. No such effects were observed in the dose group treated with 10 mg/kg bw/d. Caesarean section data obtained from the treated groups did not reveal any meaningful differences (concerning number of corpora lutea, implantation sites, viable fetuses, fetal sex and fetal weight) when compared with the controls. Fetal evaluations of type and frequency of malformations and variations did not reveal any indications for a treatment related teratogenic effect. In summary, oral administration of octadecenylamine to pregnant rats produced dose-dependent maternal toxicity in the 40 and 80 mg/kg bw/d dose groups. No indications of an embryotoxic, fetotoxic or teratogenic effect was observed at any tested level. A NOAEL/maternal toxicity of 10 mg/kg bw/d and a NOAEL/developmental toxicity of>80 mg/kg bw/d can be derived from the study.