Reaction mass of Fatty acids, montan-wax and Montan wax

Toxicity to reproduction

Administrative data

Endpoint:

multi-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable well-documented publication which meets basic scientific principles (similar to OECD guideline 416 with some deviations)

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1997

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 416 (Two-Generation Reproduction Toxicity Study)

Deviations:

yes

Remarks:

: lower extent of examinations compared to OECD 416 (no sperm parameters, oestrus cycle examinations, time of sexual maturation of offspring)

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

142583-61-7

Cas Number:

142583-61-7

IUPAC Name:

142583-61-7

Constituent 2

Reference substance name:

-

IUPAC Name:

-

Constituent 3

Reference substance name:

Policosanol

IUPAC Name:

Policosanol

Details on test material:

octacosanol: 67%
triacontanol: 14%
hexacosanol: 8%
tetracosanol: 5%
heptacosanol: 3%
nonacosanol: < 1%
dotriacontanol: > 1%
tetratriacontanol: < 1%

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: CENPALAB, Havanna
- Age at study initiation: (P) 6-8 wks; (F) 3 wks
- Housing: one male and two females for mating, pregnant females individually
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/-2
- Humidity (%): 50 +/- 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: acacia gum/water

Details on exposure:

VEHICLE
- Concentration in vehicle: 10 mg/ml
- Amount of vehicle: 2 ml/d

Details on mating procedure:

- M/F ratio per cage: 1/2
- Length of cohabitation: 3 weeks
- Proof of pregnancy: vaginal plug / sperm in vaginal smear
- After successful mating each pregnant female was caged: individually

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Concentration was verified by gas chromatography

Duration of treatment / exposure:

from 6-8 weeks of age (F0) through three successive generations

Frequency of treatment:

daily

Details on study schedule:

- F1 parental animals not mated until 2 weeks after selected from the F1 litters.
- Selection of parents from F1 generation at weaning.
- Age at mating of the mated animals in the study: approx. 16 weeks

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0 mg/kg bw/day
Basis:
other: vehicle control

Doses / concentrations 2

Remarks:

Doses / Concentrations:
5 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 3

Remarks:

Doses / Concentrations:
50 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 4

Remarks:

Doses / Concentrations:
500 mg/kg bw/day
Basis:
actual ingested

No. of animals per sex per dose:

initial 15 males and 30 females, further matings 10 males and 20 females per group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: highest dose is 1500-fold the maximal therapeutic dose in humans (20 mg/d)

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily


BODY WEIGHT: Yes
- Time schedule for examinations: weekly

Oestrous cyclicity (parental animals):

not reported

Sperm parameters (parental animals):

not reported

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter; excess pups were killed and discarded.


PARAMETERS EXAMINED
The following parameters were examined in F1 / F2 / F3 offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities

F3 offspring: behavioural tests at day 21: righting on surface, air righting, corneal, pirmal and pain reflexes, auditory startle, visual placing


GROSS EXAMINATION OF PUPS:
yes,

GROSS EXAMINATION OF DEAD PUPS:
not stated

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All animals (time point not stated)
- Maternal animals: All animals (time point not stated)


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations (not stated in detail)


HISTOPATHOLOGY / ORGAN WEIGHTS
Histopathology was performed if organs revealed abnormalitites

Postmortem examinations (offspring):

SACRIFICE
- The F1a and F2a offspring was not selected as parental animals and sacrificed after weaning.
- all animals were subjected to postmortem examinations (macroscopic and microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations (not stated in detail)


HISTOPATHOLOGY / ORGAN WEIGTHS
Histopathology was performed if organs revealed abnormalitites

Statistics:

ANOVA was used for statistical analysis of body weights, the Kruskal-Wallis test for analysis of pup numbers (live or stillborn, survivors at different time points) and duration of pregnancy, and the Fisher's exact probability test for fertility, gestation, viability and lactation indices as well as sex ratio.

Reproductive indices:

fertility index: pregancies/matings
gestation: females with live pups/females pregant

Offspring viability indices:

viability index: live pups (day 4)/live pups (day 0)
lactation index: live pups (day 21)/live pups (day 4)

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:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Effect levels (P0)

1

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: overall effects

2

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: overall effects

Remarks on result:

other: Generation: F3a/F3b (migrated information)

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Details on results (F1)

OTHER FINDINGS (OFFSPRING): the sex ratio was unaffected by the treatment. No effects were observed in the behavioural tests performed in the F3b generation at postnatal day 21

Effect levels (F1)

1

Dose descriptor:

NOAEL

Generation:

F1a

Effect level:

500 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: overall effects

2

Dose descriptor:

NOAEL

Generation:

F1b

Effect level:

500 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: overall effects

Results: F2 generation

Effect levels (F2)

Dose descriptor:

NOAEL

Generation:

F2a

Effect level:

500 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: overall effects

Overall reproductive toxicity

Reproductive effects observed:

not specified

Any other information on results incl. tables

No significant substance-related effects were observed in this study with respect to parental body weight gain, clinical signs of toxicity, organ toxicity or reproductive performance. The treatment up to the highest dose was also not affecting the offspring. No adverse effects were observed with regard to body weight gain, viability, sexual maturation, organ toxicity and neurobehavioural alterations.

Applicant's summary and conclusion

Conclusions:

Under the conditions of this 2-generation reproductive toxicity study, the test item Policosanol did not produce any deleterious effect on fertility, reproduction and development of offspring in a multigeneration study in rats at daily doses up to 500 mg/kg bw/day.

Executive summary:

The test substance Policosanol was administered to Sprague-Dawley rats in doses from 0, 5, 50 and 500 mg/kg bw/day for three consecutive generations. Two litters were reared for each generation until at least 3 weeks of age. Body weight gain was unaffected at all doses in parents and offspring. There were no clinical signs of toxicity or other substance-related toxic effects. There were no significant differences in the reproductive performance of parental animals and development of offspring (fertility, litter size, number of stillborns, sex ratio of offspring, pup survival and body weight gain, behavioural tests of the F3b generation).