Fatty acids, C18-36, esters with ethylene glycol

Toxicity to reproduction

Administrative data

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

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:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPPTS 870.3800 (Reproduction and Fertility Effects)

Deviations:

yes

Remarks:

only exposure of the parental (P0) generation

GLP compliance:

yes

Remarks:

conducted under the Cuban Good Laboratory Practices (GLP) guidelines

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Octacosanoic acid

EC Number:

208-041-8

EC Name:

Octacosanoic acid

Cas Number:

506-48-9

Molecular formula:

C28H56O2

IUPAC Name:

octacosanoic acid

Details on test material:

D-003 is a mixture of long-chain fatty acids isolated and purified from sugar cane wax whose major component is 1-octacosanoic acid, followed by 1-triacontanoic, 1-dotriacontanoic, and 1-tetratriacontanoic acids. Other acids are present at lower concentrations

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CENPALAB (La Habana, Cuba)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: (P) 10-12 wks; (F1) postnatal 10-12 wks
- Weight at study initiation: (P) Males: 123 g; Females: 98 g; (F1) no data
- Housing: individually
- Diet: CENPALAB-certified Rat lab chow ad libitum
- Water: tap water ad libitum
- Acclimation period: no data
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): 50 %
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: suspension of water with 1% Acacia Gum

Remarks:

maximum dosing volume 10 mL/kg bw

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: Dosing suspensions were prepared fresh weekly and refrigerated after corroborating their stability in such conditions.

Details on mating procedure:

- M/F ratio per cage: P0 2 females/ 1 male; F1 1 male/afemale
- Length of cohabitation: no data
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

Female rats for 15 days prior to mating, through mating and gestation to day 21 of lactation and male rats for 4 weeks prior and during mating

Frequency of treatment:

daily

Details on study schedule:

- F1 parental animals not mated until 10-12 weeks after selected from the F1 litters.
- Selection of parents from F1 generation at weaning, when pups were 21 days of age.
- Age at mating of the mated animals in the study:10-12 weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

500 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

1 000 mg/kg bw/day

No. of animals per sex per dose:

30 females and 15 males

Control animals:

yes, concurrent vehicle

Details on study design:

Only exposure of the P0 animals
Dose selection rationale: 1000 mg/kg bw is the acceptable upper limit dose by ICH/OECD guidelines.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes daily

DETAILED CLINICAL OBSERVATIONS: no data

BODY WEIGHT: Yes regulary at least on day 0, 6, 13 and 20 of gestation and at day 1, 4, 7, 14 and 21 of lactation

FOOD CONSUMPTION: Yes, regulary

WATER CONSUMPTION: No

Oestrous cyclicity (parental animals):

not investigated

Sperm parameters (parental animals):

not investigated

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 4/sex/litter excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 / F2 offspring: post natal day 1 number, weight, external examination and sex
F1 offspring: survival, weight and locomotor activity on day 4, 7, 14 and 21 (for F1 animals selected for mating: body weight weekly)
F1 offspring: day 4 unfolding of the pinnae and righting in air on day 4; day 10 hair growth; day 12 eruption of the incisors; day 13 opening of the ear; day 14 visual placing and auditory startle; day 15 opening of the eyes ; day 17 pupilar, corneal and parpebral reflexes; day 21 activity measurements
such as squares entered, urination/defecation and standing on hindlegs; day 25 testicular descent; day 37 vaginal canalization

GROSS EXAMINATION OF DEAD PUPS: no data

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY: not investigated

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY: not investigated

Postmortem examinations (parental animals):

not specified

Postmortem examinations (offspring):

not specified

Statistics:

Maternal initial body weight, maternal weight gain, fetal and pup body weights were analyzed using a parametric analysis of variance followed by Tukey's multiple comparison test. Data on dead and alive fetuses, sex ratio, litter size, survival through the weaning period and percentage of pups showing physical and reflex development were analyzed by the Kruskal–Wallis (nonparametric) test followed by Mann–Whitney U-test to determine which treatment groups differed from the control.
Trend analysis via ANOVA, Jonckheere's test and the Cochran–Armitage test.

Reproductive indices:

not specified

Offspring viability indices:

not specified

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Mortality:

not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

see attached table

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

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not examined

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Description (incidence and severity):

see attached table
Pregnancy rate: 30/30. 28/30 and 29/30 at 0, 500 and 1000 mg/kg bw

Details on results (P0)

see table

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: absence of effects

Target system / organ toxicity (P0)

Critical effects observed:

no

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

see figure

Food consumption and compound intake (if feeding study):

not specified

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

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not specified

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

not specified

Effect levels (P1)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Remarks on result:

other: no details except for body weight

Target system / organ toxicity (P1)

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

see table (live pups and dead pups)

Body weight and weight changes:

no effects observed

Description (incidence and severity):

see table

Food consumption and compound intake (if feeding study):

not specified

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:

no effects observed

Description (incidence and severity):

see table (no effects on testes and vaginal opening)

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Other effects:

no effects observed

Description (incidence and severity):

no effects on behavior and development (see table)

Developmental neurotoxicity (F1)

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

see table

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Remarks on result:

other: no effects observed

Target system / organ toxicity (F1)

Critical effects observed:

no

Results: F2 generation

General toxicity (F2)

Mortality / viability:

not specified

Description (incidence and severity):

no treatment related effects

Body weight and weight changes:

no effects observed

Description (incidence and severity):

body weight on day 1 did not show any treatment related effects

Gross pathological findings:

no effects observed

Description (incidence and severity):

no treatment related effects

Details on results (F2)

There were no treatment-related effects on mortality, weight, or external morphology of the F2 pups

Effect levels (F2)

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Remarks on result:

other: absence of effects on mortality, body weight and external examinations

Target system / organ toxicity (F2)

Critical effects observed:

no

Overall reproductive toxicity

Reproductive effects observed:

no

Applicant's summary and conclusion

Conclusions:

No effects on reproductive performance in the exposed parental generation were observed at 1000 mg/kg bw.
No effects on development and reproductive performance of the F1 generation were observed at 1000 mg/kg bw.

Executive summary:

D-003 is a mixture of long-chain fatty acids isolated and purified from sugar cane wax with cholesterol-lowering properties. D003 given orally (500 and 1000 mg/kg/day) to female rats for 15 days prior to mating, through mating and gestation to day 21 of lactation and male rats for 4 weeks prior and during mating did not induce toxic effects on reproduction. There were no significant reductions in the number of animals that conceived, in the numbers of pups born to those that did conceive, in the numbers of pups that survived until weaning, and in their body weights at weaning. No treatment related effects in offspring were observed related to growth, physical and behavioral development, spontaneous activity and reproductive performance.

The NOAEL is 1000 mg/kg bw.