Glycerides, C16-18 mono-, di- and tri-, hydrogenated, citrates, potassium salts

Toxicity to reproduction

Administrative data

Endpoint:

fertility, other

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: see 'Remark'

Remarks:

GLP guideline study with acceptable restrictions. Report does not contain all study details. Female fertility data are identical in rats and mice, indicating a transcription error. Information is used in a read-across approach. For justification of read-across and for further details please refer to the read-across report.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD 408

Principles of method if other than guideline:

Within a 90 day oral feeding study performed equivalent to OECD guideline 408 with Castor oil, male and female fertility parameters were analyzed in rats and mice. No matings were performed.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Castor oil

EC Number:

232-293-8

EC Name:

Castor oil

Cas Number:

8001-79-4

IUPAC Name:

8001-79-4

Details on test material:

- Name of test material (as cited in study report): Castor Oil
- Synonyms: Ricinus Oil, oil of Palma Christi, tangantangan oil, phorboyl, Neoloid
- Composition of test material, percentage of components: Triglyceride of fatty acids. Fatty acid composition is approximately 87% ricinoleic,
7% oleic, 3% linoleic, 2% palmitic, 1% stearic, and trace amounts of dihydroxystearic.
- Analytical grade: USP AA grade
- Source: Cas Chemical, Inc. (Bayonne, NJ, USA)
- Stability: The stability of the study material during the toxicology studies was monitored by determination of peroxide content and by high performance liquid chromatography. No deterioration of the castor oil study material was observed over the course of the studies.

Test animals

Species:

other: rats and mice

Strain:

other: F344/N and B6C3F1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Simonsen Laboratories (Gilroy, CA, USA)
- Age at study initiation: 6 weeks
- Weight at study initiation: male rats: 126 - 132 g; female rats: 107- 110 g, male mice: 22.6 - 23.0 g, female mice: 17.2 - 17.7 g
- Fasting period before study:
- Housing: rats: 5 per cage, mice individually in Polycarbonate cages lined with heat-treated hardwood chips, covered with polyester filter sheets.
- Diet (e.g. ad libitum): Control feed (NIH 07) or diet formulations of castor oil were available ad libitum; feeders were changed twice per week throughout the study.
- Water (e.g. ad libitum): automatic watering system
- Acclimation period: 14 days


ENVIRONMENTAL CONDITIONS
- Temperature (°F): 68-76°F
- Humidity (%): 42% to 72%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
Formulated diets were prepared by blending the appropriate amount of castor oil with a small quantity of feed to prepare a premix. The premix then was layered between the required amount of feed in a twin-shell blender and blended for 15 minutes to achieve a uniform mix. The homogeneity of castor oil in feed at 10% (100 mg/g) was determined by gravimetric analysis, and blends at 0.5% (5 mg/g) were determined by HPLC analysis. These concentrations of chemical in feed were found to be homogeneously distributed by this mixing procedure. The stability of the 0.5% dose level was determined using HPLC; it was 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 a rodent cage. During the studies, formulated diets were stored for no longer than 3 weeks at 5°C; feed hoppers in the animal cages were changed twice weekly.

Details on mating procedure:

no matings performed

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The results of the analyses for all dose mixtures given to the animals ranged from 97% to 106% of the target concentrations.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

daily ad libitum feeding

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0, 2.50, 5.00, 10.0 % (w/w)
Basis:
nominal in diet

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0, 1583, 3067 and 5835 mg/kg bw/day
Basis:
other: actual ingested: male rats

Doses / concentrations 3

Remarks:

Doses / Concentrations:
0, 1569, 3045, 5725 mg/kg bw/day
Basis:
other: actual ingested: female rats

Doses / concentrations 4

Remarks:

Doses / Concentrations:
0, 3800, 7823, 15017 mg/kg bw/day
Basis:
other: actual ingested: male mice

Doses / concentrations 5

Remarks:

Doses / Concentrations:
0, 5009, 9627, 16786 mg/kg bw/day
Basis:
other: actual ingested: female mice

No. of animals per sex per dose:

10

Control animals:

yes, plain diet

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS/ CLINICAL OBSERVATIONS: Yes
- Time schedule: twice a day

BODY WEIGHT: Yes
- Time schedule for examinations: initially and 1 x wk thereafter.


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: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

Oestrous cyclicity (parental animals):

Proestrus, Estrous, Metestrous, Diestrous, Cycle Length (days)

Sperm parameters (parental animals):

Caudal weight, epididymal weight, testis weight, Sperm count/g testis, Sperm motility (%)

Litter observations:

not performed

Postmortem examinations (parental animals):

Complete histopathology examinations were conducted on all rats and mice from the control and 10% dose groups. Livers were examined from male rats in all other dose groups; histologic sections of gross lesions were examined from all rats. Organ weights were determined to the nearest milligram for the liver, right kidney, right testicle, heart, thymus, and lungs. All tissues were preserved in 10% neutral buffered formalin.

The following tissues were routinely processed for preparation of histologic sections and microscopic examination: adrenal
glands, brain, cecum, colon, duodenum, epididymis/seminal vesicles/prostate/testes or ovaries/uterus, esophagus, eyes (if
grossly abnormal), femur (including marrow), heart, ileum, jejunum, kidneys, liver, lungs and mainstem bronchi, mammary
gland, mandibular and mesenteric lymph nodes, nasal cavity and turbinates, pancreas, parathyroid glands, pituitary gland,
preputial or clitoral glands, rectum, salivary glands, skin, spinal cord and sciatic nerve (if neurologic signs present), spleen,
forestomach and glandular stomach, thymus, thyroid gland, trachea, urinary bladder, zymbal glands, and all gross lesions
and tissue masses including regional lymph nodes. A complete histopathologic examination was conducted on all rats and
mice from the control and 10% dose groups. Liver was examined from male rats in all other dose groups, and histologic
sections of gross lesions were examined from all rats.

Postmortem examinations (offspring):

not performed

Statistics:

Dunn's test; Shirley's test.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

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:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

not examined

Details on results (P0)

CLINICAL SIGNS AND MORTALITY
No effects

BODY WEIGHT AND WEIGHT GAIN
Group mean body weights of rats receiving diets containing castor oil did not differ significantly from controls. Mean body weights of exposed female rats were slightly lower than the mean body weights of controls but the differences were not dose-related.
Mean body weights of exposed male mice generally were lower than controls, while mean body weights of exposed females generally were higher. There were no obvious indications that these differences were related to dietary concentrations of castor oil, except that mean body weights of male mice receiving the 10% castor oil diet were consistently lower than those of control mice from week 3 through the end of the study.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
No significant differences in average food consumption among each sex were observed, although food consumption of male and female rats receiving diets containing 10% castor oil was slightly lower than that of controls. Similarly, food consumption by female mice receiving diets containing 10% castor oil was slightly lower than controls.

ORGAN WEIGHTS
In male rats, there was a slight decrease in epididymal weight (6-7%) which occurred in the middle- and high-dose groups, but this was not dose-related. There were no effects on any other male rat reproductive endpoint, or on any female rat reproductive endpoint. Although there was
some variation in epididymal weights, their small magnitude and the absence of changes in other endpoints suggested that there was little or no evidence of any reproductive toxicity associated with castor oil exposure. Histopathologic examination revealed an absence of compound-related lesions in any organ or tissue of rats exposed to castor oil in the diet.

Castor oil exposure produced no adverse effects on any male (testes weight, epididymal sperm motility, density, or testicular spermatid head count) or female (estrual cycle length, or time spent in each phase of the cycle) reproductive parameter among mice. The low value for sperm motility
in control mice was attributed to poor preparative technique.

Effect levels (P0)

1

Dose descriptor:

NOAEL

Remarks:

parental fertility parameters

Effect level:

ca. 5 000 mg/kg bw/day (actual dose received)

Based on:

other: calculated test material intake based on food consumption and body weight

Sex:

male/female

Basis for effect level:

other: for rats based on oestrus stage and cycle length and sperm characterization.

2

Dose descriptor:

NOAEL

Remarks:

parental fertility parameters

Effect level:

ca. 15 000 mg/kg bw/day (actual dose received)

Based on:

other: calculated test material intake based on food consumption and body weight

Sex:

male/female

Basis for effect level:

other: for mice based on oestrus stage and cycle length and sperm characterization.

Results: F1 generation

General toxicity (F1)

Clinical signs:

not examined

Mortality / viability:

not examined

Body weight and weight changes:

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

Details on results (F1)

not examined

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Table 1: Reproductive System Data for F344/N Rats in the 13-Week Feed Studies

of Castor Oil

 

	Percent in Feed
	0	2.5	5	10
Male^a
Left caudal weight (mg)	151	153	145	153
Left epididymal weight (mg)	502	498	464*	476
Left testis (mg)	1539	1550	1463	1492
Sperm count (x10^6)/g testis	72.8	65.9	71.7	77.5
Sperm motility (%)	73.6	65.9	72.1	69.8
Female^b
Estrous stage (%)
Proestrus	12.5	14.2	15.8	16.7
Estrous	28.3	32.5	25.8	25.8
Metestrous	18.3	19.2	18.3	19.2
Diestrous	40.8	34.2	39.2	38.3
Not clear or no cells observed	0.0	0.0	0.8	0.0
Cycle Length (days)	5.0	5.1	5.2	5.1

^aMean for groups of 10 animals; no significant difference vs. the controls by Dunn's test

^bMean for groups of 10 animals unless otherwise specified

* Significantly different from control groups by Shirley's test ; p < 0.05.

 

Table 2: Reproductive System Data for B6C3F1 Mice in the 13-Week Feed Studies

of Castor Oil

 

	Percent in Feed
	0	2.5	5	10
Male^a
Left caudal weight (mg)	15	13	16	16
Left epididymal weight (mg)	45	46	46	44
Left testis (mg)	121	120	121	119
Sperm count (x106)/g testis	179.2	162.4	170.1	158.3
Sperm motility (%)	39.2	53.7	45.4	52.2
Female^b
Estrous stage (%)
Proestrus	12.5	14.2	15.8	16.7
Estrous	28.3	32.5	25.8	25.8
Metestrous	18.3	19.2	18.3	19.2
Diestrous	40.8	34.2	39.2	38.3
Not clear or no cells observed	0.0	0.0	0.8	0.0
Cycle Length (days)	5.0	5.1	5.2	5.1

^aMean for groups of 10 animals; no significant difference vs. the controls by Dunn's test

^bMean for groups of 10 animals unless otherwise specified

 

Applicant's summary and conclusion

Conclusions:

A NOAEL of 5000 mg/kg bw/day for rats and a NOAEL of 15000 mg/kg bw/day for mice could be identified based on parental fertility parameters.
Matings were not performed in this study.

Executive summary:

In a subchronic toxicity study castor oil (CAS 8001-79-4) was administered to 10 rats & 10 mice/sex/dose in the diet at dose levels up to 10%.

To screen for potential reproductive toxicity, sperm motility and morphology were evaluated at necropsy, and vaginal cytology was evaluated on core-study animals during the week just preceding necropsy. For the 12 days prior to termination, females were subject to vaginal lavage with saline. The relative preponderance of leukocytes, nucleated epithelial cells, and large squamous epithelial cells were used to identify the stages of the estrual cycle. Sperm motility and sperm density was evaluated at necropsy, as well as the number of spermatid heads per total testis and per gram of testis. No adverse effects have been reported at any dose. The NOAEL is 5000 mg/kg/d (rat) and 15.000 mg/kg/d (mice).

This subchronic toxicity study in the rat/mouse is acceptable and satisfies the guideline requirement for a subchronic oral study (OECD 408).