Glycerides, C16-18

Toxicity to reproduction

Administrative data

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not available

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

Long-term and short-term effects of dietary cholesterol and fats in the Mongolian gerbil

Author:

Temmerman AM, Vonk RJ, Niezen-Koning K, Berger R and Fernandes J

Year:

1988

Bibliographic source:

Ann. Nutr. Metab. 11:177-185

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

A two generation reproductive toxicity study was conducted in Mongolian gerbils to assess the effects of dietary exposure to 8.75% w/w palm kernel oil (PO).
5 couples of adult gerbils with their sucklings (generation 1) were assigned to various dietary groups: palm kernel oil (8.75% w/w), sunflower seed oil (8.75% w/w) or cholesterol (0.2% w/w). The ensuing generations were put on the same diets as their parents with the exception of the cholesterol-containing diets which was reduced to 0.05%. After 4 months, 4 adults in each group were terminated. and histopathological changes in liver and spleen were observed along with liver and serum cholesterol levels. In the second generation, parameters such as frequency of litters, number of pups, mean litter size, mean weight, postnatal mortality were also observed.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Glycerides, C8-18 and C18-unsatd.

EC Number:

266-946-3

EC Name:

Glycerides, C8-18 and C18-unsatd.

Cas Number:

67701-28-4

Molecular formula:

Triglycerides containing a glycerol backbone esterified to fatty acids with a carbon chain length of C8-18 as well as unsaturated C18 fatty acids

IUPAC Name:

Tri(C8-18 and C18-unsatd. fatty acyl)glycerol

Details on test material:

- Name of test material (as cited in study report): Palm kernel oil (CAS N° 8023-79-8, EC N° 232-425-4). Under the SDA nomenclature, the name of this substance is 'Glycerides, C8-18 and C18-unsatd.'

Test animals

Species:

other: gerbil

Strain:

other: Mongolian

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: Adult gerbils and their sucklings (F1) were used at the initiation
- Housing: Housed in couples in standard laboratory cages with a wire mesh lid with food hopper. The bedding consisted of wood shavings.
- Diet (e.g. ad libitum): Diets manufactured by Hope Farms BV (Woerden, The Netherlands) were used. The test diet contained increased protein (26 %), biotin, choline, inositol and vitamin E (50 %) and, cellulose (1 %).
- Water (e.g. ad libitum): Water acidified with HCI (pH 3), ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature: 20 °C
- Humidity: 50 %
- Photoperiod: 10 hrs light/14 hrs dark

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

Not reported

Details on mating procedure:

No data

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Not applicable

Duration of treatment / exposure:

2 generations (specific period not mentioned)

Frequency of treatment:

Daily

Details on study schedule:

Not reported

Doses / concentrations

Dose / conc.:

8.75 other: % w/w in basic diet

Remarks:

Basis: nominal conc.

No. of animals per sex per dose:

5 couples per group

Control animals:

yes, plain diet

Details on study design:

No data

Positive control:

Not applicable

Examinations

Parental animals: Observations and examinations:

Not examined

Oestrous cyclicity (parental animals):

Not examined

Sperm parameters (parental animals):

Not examined

Litter observations:

PARAMETERS EXAMINED
The following parameters were examined in [F2] offspring: Frequency of litters, number of pups, mean litter size, mean weight at 6 months of age and postnatal mortality

Postmortem examinations (parental animals):

Not examined

Postmortem examinations (offspring):

SACRIFICE
- Four adult animals out of each dietary group of the F1 generation were sacrificed after 4 months.

HISTOPATHOLOGY / ORGAN WEIGHTS
- Tissues/organs examined: Liver and spleen
- Brief description on slide preparation: After fixation and paraffin embedding of parts of the liver and spleen, 4-µm thick sections were cut for analysis, followed by staining with hemaloxylin, eosin and Gomori's reticulin.

CLINICAL CHEMISTRY
- Total serum cholesterol and liver cholesterol content were also determined along with histopathological parameters in F1 generation after sacrifice

Statistics:

The Student-t test for unpaired samples was used to compare the variables of the different groups. For large differences in variance between the samples, the Wilcoxon test for unpaired samples was used. p < 0.05 was considered significant.

Reproductive indices:

Not reported

Offspring viability indices:

Not reported

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

not examined

Body weight and weight changes:

not examined

Food consumption and compound intake (if feeding study):

not examined

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

not examined

Other effects:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

not examined

Details on results (P0)

No data

Effect levels (P0)

Key result

Dose descriptor:

NOAEL

Effect level:

8.75 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant treatment-related effect

Target system / organ toxicity (P0)

Key result

Critical effects observed:

no

Results: P1 (second parental generation)

Effect levels (P1)

Key result

Dose descriptor:

NOAEL

Effect level:

8.75 other: % w/w in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant treatment-related effect

Target system / organ toxicity (P1)

Key result

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

not examined

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Histopathological findings:

no effects observed

Details on results (F1)

BODY WEIGHT (OFFSPRING): No significant difference compared to controls was observed (F1 and F2)


HISTOPATHOLOGY (F1): The livers and spleens of the animals on diets basal diet and palm kernel oil diet had a normal histology.


CLINICAL CHEMISTRY (F1):
- Serum cholesterol (mmol/L): Slight increase in PO fed group (3.9±0.5) in comparison with control (2.8±0.3).
- Liver cholesterol content: No significant difference was observed.


FREQUENCY OF LITTERS (F2): No significant effects were observed.


NUMBER OF PUPS (F2): No significant difference was observed (PO: 116 and basal diet (control): 146).


MEAN LITTER SIZE (F2): No significant difference was observed (PO: 4.7±1.7 and basal diet (control): 4.9±1.6).


POSTNATAL MORTALITY (F2): No significant mortality was observed in comparision to basal diet group ( PO: 4 % and basal diet (control): 3 %)

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

8.75 other: % in diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no significant treatment-related effect

Target system / organ toxicity (F1)

Key result

Critical effects observed:

no

Results: F2 generation

Effect levels (F2)

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

> 8.75 other: % w/w

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect on frequency of litters, mean litter size, total no. of newborns and no suckling death

Target system / organ toxicity (F2)

Key result

Critical effects observed:

no

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Applicant's summary and conclusion

Conclusions:

Under the conditions of the two generation study, the test substance exhibited no significant reproductive toxicity at 8.57% in diet.

Executive summary:

A two generation reproductive toxicity study was conducted in Mongolian gerbils to assess the effect of ‘glycerides, C8-18 and C18-unsatd.’ (as palm kernel oil). Adult Mongolian gerbils (highly resistant to atherosclerosis) and their sucklings (first generation) were randomly assigned to two groups of 5 couples each (group 1: basal diet; group 2: 8.75% test substance in diet). Four months after starting the diets, four adult animals of each dietary group of the first generation were sacrificed and histopathological changes in liver and spleen were observed along with liver and serum cholesterol levels. In the second generation parameters such as frequency of litters, number of pups, mean litter size, mean weight, postnatal mortality were also observed. Both in the first and second generations, no significant differences of body weight were observed compared to controls. The liver and spleen of the animals of the first generation also had a normal histology. However, a slightly elevated serum cholesterol level was observed in the test substance fed group. No significant effect on frequency of litters, mean litter size, total no. of newborns, and no suckling death were found in the second generation of the test substance fed group compared to the basal dietary group. Hence, under the test conditions, the substance did not exhibit any significant reproductive toxicity at 8.57% in diet (Temmerman, 1988).