Nonanoic acid

Toxicity to reproduction

Administrative data

Endpoint:

three-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

significant methodological deficiencies

Remarks:

Not the recommended number of animals. histopathology only on selected organs. No information on GLP status, limited investigation depth

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1993

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Deviations:

yes

Remarks:

3 generations; lower animal numbers; histopathology only on selected organs; No information on GLP status

GLP compliance:

not specified

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

cuphea oil

IUPAC Name:

cuphea oil

Constituent 2

Reference substance name:

Cuphea oil

IUPAC Name:

Cuphea oil

Details on test material:

- Name of test material (as cited in study report): cuphea oil
- Molecular formula (if other than submission substance): triglyceride
- Fatty acid composition:
octanoate 4.8%
decanoate 75.9%
dodecanoate 2.5%
myristate 2.2%
palmitate 3.4%
stearate 0.7%
oleate 3.3%
linoleate 5.5%h
- Composition of test material, percentage of components: see above; analysed using gas chromatography

Test animals

Species:

mouse

Strain:

other: two strains were tested: CBA/2 and C57B1/6

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SASCO (Omaha, NE)
- Age at study initiation: (P) 8-10 wks
- Housing:
P: two females and one male together; females singly when vaginal plug was observed
F1, F2: weaned at 4 weeks, then housed together until sexual maturation, mating two females: one male. Females tehn hosued singly.
Males of each generation housed singly an dfed for 13 weeks; some males were fed for 5-12 months

- Diet: ad libitum
Feeding: Cuphea oil was limited, schedule was therefore as follows:
Strain 57B1/6:
F1: 10 months
F2: 8 months
F3: 11-12 months

Strain CBA/2:
F1: 11-12 months
F2: 9-11 months
F3: 6-8 months

- Water: ad libitum

Administration / exposure

Route of administration:

oral: feed

Vehicle:

other: basal diet

Details on exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): monthly
- Mixing appropriate amounts with (Type of food): basal diet with reduced beef tallow 886 g/kg instead of 172g/kg
- Storage temperature of food: 4°C

Details on mating procedure:

- M/F ratio per cage: 1:2
- Length of cohabitation: until vaginary plug was noted
- Proof of pregnancy: vaginal plug
- After successful mating each pregnant female was caged: singly

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

GC determination of fatty acids in Cuphea oil

Duration of treatment / exposure:

Feeding: Cuphea oil was limited, schedule was therefore as foollows:
Strain 57B1/6:
F1: 10 months
F2: 8 months
F3: 11-12 months

Strain CBA/2:
F1: 11-12 months
F2: 9-11 months
F3: 6-8 months

Frequency of treatment:

7 days/week

Doses / concentrations

Remarks:

Doses / Concentrations:
8.6%
Basis:
nominal in diet

No. of animals per sex per dose:

Strain 57B1/6: F1: 16; F2: 16; F3: 13. Controls 15-17/generation

Strain CBA/2: F1: 25; F2: 11; F3: 5. Controls 5-29/generation

Control animals:

yes

Positive control:

no

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations:
Animals fed for 13 weeks: weeks 4, 8, and 13
Animals fed for 5-12 months: : weeks 4, 13. 26, 39, and 45


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. Total food intake in grams is given.
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No.

OTHER:

Oestrous cyclicity (parental animals):

No data

Sperm parameters (parental animals):

No data

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no data

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
number of live births, postnatal mortality, presence of gross anomalies, weight gain

GROSS EXAMINATION OF DEAD PUPS:
no

Postmortem examinations (parental animals):

no data

Postmortem examinations (offspring):

no data

Statistics:

no data

Reproductive indices:

pregnancy index, pubs born/female

Offspring viability indices:

pubs at weaning/pups born

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

Description (incidence and severity):

no consistent variation, but cuphea-fed mice had slightly lower body weights and lower feed intake

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

no consistent variations, but cuphea-fed mice had slightly lower body weights and lower feed intake

Food efficiency:

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Liver: excessive lipid accumulation (liver of most mice).
Kidneys: fat vacuoles in proximal cortical tubules of all mice; spleen in some mice enlarged

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Description (incidence and severity):

Test substance intake: no consistent variations, but cuphea-fed mice had slightly lower body weights and lower feed intake

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Effect levels (P0)

Key result

Dose descriptor:

NOAEL

Effect level:

8.6 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

no effects observed

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

CBA/2 mice produced fewer offspring than females fed the basal diet.

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

no effect in C57B1/6, in contrast to strain CBA/2 (strain-specific effect). Viability of pups from F1 and F2 parents reduced compared to those from F0 pups, but independent of the diet

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

10% reduced body weights in Cuphea fed C57B1/6 pups after 8 and 13 weeks. Reduced body weights in Cuphea fed CBA/2 pups after 13 weeks

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

18% reduced food intake in cuphea fed C57B1/6 mice after week 8. Reduced food intake in cuphea fed CBA/2 pups after 8 and 13 weeks.

Food efficiency:

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

no effects observed

Anogenital distance (AGD):

not specified

Nipple retention in male pups:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

comparable between grouops; where statistically significant changes were noted, Cuphea-fed mcie had lower body and liver weights

Gross pathological findings:

not specified

Histopathological findings:

effects observed, treatment-related

Description (incidence and severity):

Liver: excessive lipid accumulation (liver of most mice); Kidneys: fat vacuoles in proxicmal cortical tubules of all mice; spleen in some mice enlarged

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

8.6 other: %

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Results: F2 generation

General toxicity (F2)

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

In C57B1/6 mice, body weights in Cuphea fed pups were decreased by 20% at four weeks of age.
Reduced body weights in Cuphea fed CBA/2 pups after 8 weeks.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Reduced food intake in Cuphea fed CBA/2 pups after 8 weeks

Developmental neurotoxicity (F2)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not specified

Effect levels (F2)

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

ca. 8.6 other: %

Sex:

male/female

Basis for effect level:

other:

Remarks on result:

not determinable due to absence of adverse toxic effects

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

Comments to histopathology in F0, F1, and F3 animals of both tested strains:

Liver: lipid accumulation due to fat and cholesterol content of the diets. No differences between diets and generations in fatty liver development. The two strains showed marked differences, possibly because of differences in hepatic fat metabolism.

Renal vacuolar changes: likely caused by the high total lipid content of the fee No differences between diets or generations (table 7).

Preputial glands: strain-specific differences, no effect of treatment.

Lungs:. Changes noted, but not related to treatment or strain.

 

 

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:

Feeding Cuphea oil to mice (two strains, 3 generations each) did not affect reproduction and had no adverse effect on parental animals. Cuphea oil appears to be nontoxic, but was of no particular health benefit. The NOAEL according to this study is > 8.6% cuphea oil in feed.

Executive summary:

In this 3 generation feeding study two strains of mice (CBA/2 and C57B1/6) were fed a diet of 8.6% cuphea oil. Male and female mice were used. Controls were fed a basal diet with 17.2% beef tallow and 3.5% corn oil; in the diet for treated mice. In the treated mice, 8.6% Cuphea oil replaced 50% of the beef tallow, i.e. the diets were essentially isocaloric. The triglycerides of the Cuphea oil contained 75% decanoic acid, 4.8% octanoic acid, and small proportions of some long chain fatty acids.

Feeding Cuphea oil to mice (two strains, 3 generations each) did not affect reproduction and had no adverse effect on parental animals. Failureof mothers to succesfully nurture pups to weanling age (four weeks) was the only reproductive problem noted and affected the F1 and F2 mice of both diets, thus this is not related to cuphea oil. The F0 mice were exposed to a different diet and environment than were the F1 and F2 moce before breeding, but it is not clear how these differences might have led to the impairment of survival of pups in the later generations.

Animals were treated up to 12 months. Thus, Cuphea oil appears to be nontoxic, but was of no particular health benefit.

The study is not comparable with current test guidelines. However, basic scientific principles are met. According to this study, the NOAEL is 8.6% cuphea oil in both strains of mice.