Hydroxyoctadecanoic acid, monoester with glycerol

Toxicity to reproduction

Some information in this page has been claimed confidential.

Administrative data

Endpoint:

fertility, other

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 1988-July 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: Fertility measures added to a 13 week repeated dose toxicity study

Principles of method if other than guideline:

Fertility measures and pathology assessment of reproductive organs added to a guideline 13-week repeated dose toxicity study

GLP compliance:

not specified

Remarks:

Peer reviewed

Limit test:

no

Test material

Test material information

Constituent 1

Test material form:

liquid: viscous

Details on test material:

Castor oil (CAS 8001-79-4), lot #L-5G30-01, was obtained from Cas Chemical, Inc., Bayonne, NJ. Purity analysis indicated that it was consistent with the USP specifications and the reported composition for castor oil: Analysis was conducted by Midwest Research Institute (MRI), in Kansas City, MO, utilizing infrared, UV/Vis and NMR spectroscopy, Karl Fischer water analysis, TLC and HPLC, and a battery of USP standard analyses for castor oil.
The stability of the test material during the toxicology studies was monitored by determination of peroxide content and by HPLC. The substance was stable during the course of the studies.

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

Simonsen Laboratories, Gilroy, CA
6 weeks of age
held 14-15 days in quaranteen after receipt and before dosing
Rats housed 5/cage, mice housed individually
Polycarbonate with heat-treated wood chips
NIH-07 diet, ad libitum. Water ad libitum
Temp-68-76°F; relative humidity-42-72%; fluorescent light 12 h/d; 10 room air changes/h.
Animals were observed twice daily, weighed initially and weekly thereafter.
10 rats/sex/group in the core study.

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: rat chow (NIH 07 diet)

Details on exposure:

Formulated diets were prepared from a premix of castor oil with a small quantity of feed (NIH-07 diet). The premix was then blended with additional food using a twin shell blender. Homogeneity of castor oil in feed was documented by gravimetric analysis or HPLC. Diets were found to be stable for at least 21 days when stored in the dark at 5°C, and for 3 days when stored open to air and light in the rodent cages. Formulated diets were stored for no longer than 3 weeks at 5°C, and food in hoppers were changed twice weekly.

Details on mating procedure:

No mating

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Periodic analysis of castor oil-formulated diets was conducted by HPLC. All but a single sample were within specifications (± 10% of target concentration); the one sample was remixed before being given to animals. The range of analyses was 97% to 106% of target concentration.

Duration of treatment / exposure:

Food with added test material was available ad libitum, daily

Frequency of treatment:

daily

Details on study schedule:

Dosing was performed daily for 13 weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0.62 other: % in diet

Remarks:

Approximately equivalent to 400 mg/kg bw/d (nominal)

Doses / concentrations 2

Dose / conc.:

1.25 other: % in diet

Remarks:

Approx equal to 800 mg/kg bw/d (nominal)

Doses / concentrations 3

Dose / conc.:

2.5 other: % in diet

Remarks:

Approx equal to 1500 mg/kg bw/d (nominal)

Doses / concentrations 4

Dose / conc.:

5 other: % in diet

Remarks:

approximately equal to 3000 mg/kg bw/d (nominal)

Doses / concentrations 5

Dose / conc.:

10 other: % in diet

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations checked were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No. By cage, with 5 animals/cage.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

Oestrous cyclicity (parental animals):

In Females, vaginal cytology was evaluated on core-study animals during the week just preceding necropsy, using the procedure of Morrissey, et.al., 1988. For the 12 days prior to termination, females were subject to vaginal lavage with saline. The aspirated cells were air-dried onto slides, stained with Toluidine Blue O and cover slips applied. The relative preponderance of leukocytes, nucleated epithelial cells and large squamous cells were used to identify the stages of the estrual cycle.

Sperm parameters (parental animals):

In males, sperm motility and morphology were evaluated on core-study at necropsy, according to Morrissey, et.al., 1988. The left epididymis was removed and quickly weighed; the cauda epididymis was removed at the junction of the vas deferens and the corpus epididymis, then weighed. A small cut was made in the distal cauda epididymis, from which the sperm were removed, and dispersed. The number of moving and non-moving sperm were counted in 5 fields of 30 sperm or less on each slide. After sperm sampling, the cauda was placed in phosphate buffered saline (PBS) and gently chopped. The solution was mixed gently and heat fixed at 65 deg C. Sperm density was then determined using a hemocytometer.

The left testes was frozen and stored. After thawing, testicular spermatid head count was determined by removing the tunica albuginea and homogenizing in PBS containing 10% DMSO. Homogenization spermatid nuclei were counted using a hemocytometer; the data were expressed as spermatid heads per total testis and per gram of testis.

Litter observations:

not applicable

Postmortem examinations (parental animals):

The following reproductive tissues were routinely processed for preparation of histologic sections and microscopic examination: epididymis, seminal vesicles, prostate and testes in males, and ovaries and uterus in females. A complete histopathologic examination was conducted on all rats and mice from the control and 10% dose groups.

Postmortem examinations (offspring):

not applicable

Statistics:

Body weight and organ weight data were statistically analyzed within each sex by one-way Analysis of Variance tests, followed by Dunnett's t-test if pair-wise comparisons were indicated (p < 0.05)(Dunnett, 1955).

Offspring viability indices:

not applicable

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

Description (incidence and severity):

Not specifically examined, although clinical chemistry values are consistent with a high lipid diet.

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Hematological: males: statistically significant decrease in males in MCV in the 10% group, transient decrease in MCHC in males in the 10% group, decrease in the MCH in males of the 5 and 10% group, decreased platelets in the 1.25%, 5.0% and 10% groups. In females, there was a transient decrease in reticulocytes in the 0.62% and 10.0% groups. These effects were not biologically significant.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

At days 5, 21 and 90, in males and females, there was a consistent dose-related increase in serum alkaline phosphatase activity in the 5 and 10% groups. Total bile acids were also statistically significantly elevated in males of the 5 and 10% groups at 5, 21 and 90 days. This is likely an adaptation to increased absorption and metabolism of lipids from the intestinal tract.

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

not examined

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

Details on results (P0)

No toxicologically relevant adverse effects were observed in any treatment group. Absolute liver weight and liver-to-body ratio were increased in male rats at the high dose (10% group). In males, there was a slight decrease in epididymal weight (< 10%) in the middle and high dose groups, but this was not dose-related. The small magnitude and the absence of changes in other reproductive endpoints suggested that there was little or no evidence of any reproductive toxicity associated with castor oil exposure. There were no adverse effects on sperm morphology or function. There were no adverse effects on estrous cycling in females.

In males in the highest dose group (10%), there was a statistically significant decrease in MCV; this was not considered biologically significant. Also in this group, there was a slight decrease in MCH and the MCHC was transiently decreased at day 21. Other minor effects were observed in hematologic parameters: at 90 days, the platelet count in males of the 1.25, 5 and 10% groups was significantly increased. These variations were evaluated as not biologically significant by the study authors.

Serum alkaline phosphatase activity was increased in a treatment- and dose-related manner at days 5, 21 and 90, in both males and females in the 5 and 10% groups. Total bile acids were also statistically significantly elevated in males of the 5 and 10% groups at 5, 21 and 90 days. This is likely an adaptation to increased absorption and metabolism of lipids from the intestinal tract.

Histological examination revealed an absence of compound related lesions in any organ or tissue of rat exposed to castor oil in the diet.

The NOAEL was 10% in the diet, equivalent in females to 5725 mg/kg bw/d, and in males to 5835 mg/kg bw/d.

Effect levels (P0)

Key result

Dose descriptor:

NOAEL

Effect level:

6 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects were observed at the highest dose tested (10% Castor oil in the diet, equivalent to 6000 mg/kg bw/d). The NOAEL is > 6000 mg/kg bw/d.

Target system / organ toxicity (P0)

Key result

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

not examined

Mortality / viability:

not examined

Body weight and weight changes:

not examined

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

not examined

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Details on results (F1)

No F1 generation was established.

Effect levels (F1)

Key result

Dose descriptor:

other: No F1 generation established

Remarks on result:

not measured/tested

Target system / organ toxicity (F1)

Key result

Critical effects observed:

no

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Any other information on results incl. tables

There were no observed clinical effects of dietary administration of castor oil to F344 rats for 13 weeks. The administration of diets containing up to 10% castor oil was not associated with toxicity to any specific organ, organ system or tissue, including reproductive organs, under the conditions of this study. There were no effects on sperm morphology or function in males, nor on estrous cycling in females.

Actual Dosage of Test Material in Rats

Sex	Target Concentration (% in feed)	Feed Consumption (g/kg bw/d)	Test Material Consumption (mg/kg bw/d)
Males	0	65	0
	0.62	65	404
	1.25	65	809
	2.5	63	1583
	5	61	3067
	10	58	5835
Females	0	64	0
	0.62	65	401
	1.25	64	797
	2.5	63	1569
	5	61	3045
	10	57	5725

 

Applicant's summary and conclusion

Conclusions:

The effects of dietary administration of castor oil on the reproductive organs and estrous cycle of F344 rats were investigated in a U.S. NTP guideline repeated dose toxicity study. Diets containing up to 10% castor oil for 13 weeks were not associated with toxicity to reproductive organs, organ systems/ tissues, or reproductive function, under the conditions of this study. The NOAEL is greater than 10% in the diet, equivalent to 5725 mg/kg bw/d in females, 5835 mg/kg bw/d in males (actual).