O,O'-dioctadecylpentaerythritol bis(phosphite)

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 December 2005 to 6 March 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EPA OPPTS 870.3550 (Reproduction/ Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

O,O'-dioctadecylpentaerythritol bis(phosphite)

EC Number:

223-276-6

EC Name:

O,O'-dioctadecylpentaerythritol bis(phosphite)

Cas Number:

3806-34-6

Molecular formula:

C41H82O6P2

IUPAC Name:

3,9-bis(octadecyloxy)-2,4,8,10-tetraoxa-3,9-diphosphaspiro[5.5]undecane

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): 2, 4, 8, 10-Tetraoxa-3,9-diphosphasipiro[5.5]undecane, 3,9-bis(octadecyloxy)-(9Cl)
- Physical state: solid
- Storage condition of test material: refrigeration (2 - 8 °C)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Advinus Therapeutics Private Ltd., Bangalore - 560 058, India
- Age at study initiation: 12 weeks
- Weight at study initiation:
Males: Control 375 ± 17.1 g, 100 mg/kg 378 ± 16.2 g, 400 mg/kg 378 ± 18.9 g and 1000 mg/kg 377 ± 13.9 g (mean ± SD)
Females: Control 235 ± 10.4 g, 100 mg/kg 234 ± 13.7 g, 400 mg/kg 233 ± 11.9 g and 1000 mg/kg 233 ± 9.9 g (mean ± SD)
- Housing: groups of 2 per sex in sterilised suspended standard polyproylene cages (males, females during pre-mating); individually in polypropylene cages with steam sterilised paddy husk and sterilised nesting material from gestation day 20 up to lactation day 4 (females)
- Diet: Ssniff rats/mice food (Sniff Spezialdiäten GmbH., Ferdinand-Gabriel-Weg 16, D-59494 Söest, Germany) ad libitum
- Water: purified deep bore-well water
- Acclimation period: 12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 23 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 12 - 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

IN-LIFE DATES: From 31 December 1995 to 6 March 2006

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: test material suspensions in CMC were prepared daily before gavage administration. Test material was ground in a pestle and mortar before being added to 0.5 % aqueous carboxymethyl cellulose. The suspension was made up to the final volume of 90 mL to get test material concentrations of 10, 40 and 100 mg/mL.Test material was administered at an equivolume of 10 mL/kg bw.

VEHICLE
- Justification for use and choice of vehicle (if other than water): based on a solubility test, the test material was found to be insoluble in water but a clear suspension was formed in aqueous 0.5 % carboxymethyl cellulose.

Details on mating procedure:

- M/F ratio per cage: 1 male / 1 female
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

- Once daily for 2 weeks, continuing during mating period and approximately 2 weeks post mating (males)
- Once daily throughout treatment period. Treatment started 2 weeks prior to the mating period and continued through mating, pregnancy and up to lactation day 4 (females)

Frequency of treatment:

Daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 400, 1000 mg/kg
Basis:
nominal conc.

No. of animals per sex per dose:

10 males and 10 females per dose level

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: dose levels selected in consideration of results from a 7-day oral (gavage) toxicity study with 2,4,8,10-tetraoxa-3,9-diphosphaspiro [5.5]undecane, 3,9-bis(octadecyloxy)-(9Cl) in Wistar rats treated at 100, 400 and 1000 mg/kg bw/day. Under the conditions of the study no clinical signs were observed, there were no deaths and body weights, food intake, organ weights and gross pathology did not reveal any significant differences in treatment groups compared to the respective controls.
- Rationale for animal assignment (if not random): Grouping was done one day before treatment using an in-house method of bodyweight stratification and distribution. The rats procured for the study were weighed and segregated depending on bodyweight ragnes. The bodyweight randes were 331-410 for males and 201-270 g for females. Animals with bodyweight ranges 341-350, 351-360, 361-370, 371-380, 381-390 and 391-400 g for males and 211-220, 221-230, 231-240, 241-250 and 251-260 g for females were selected and rats within each bodyweight range were randomly distributed to all groups to attain groups whose mean bodyweight variation were not more than 20 %.

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily
- Cage side observations: checks for morbidity and mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily
- Cage side observations: appearance, behaviour and clinical signs. Females were also observed for signs of difficult and prolonged parturition.

BODY WEIGHT: Yes
- Time schedule for examinations: day 1 of treatment and at least weekly thereafter. All dams were weighed on gestation days 0, 7, 14 and 20 and on lactation days 0 and 4.

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes (based on weekly intakes).

Litter observations:

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

GROSS EXAMINATION OF DEAD PUPS: yes, all dead and sacrificed pups were examined for malformations and subjected to gross pathological examination

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals following a minimum treatment period of 4 weeks (this included a minimum of 2 weeks prior to mating, during the mating period and approximately 2 weeks post mating)
- Maternal animals: All surviving animals following a minimum treatment period of 4 weeks (this included at least 2 weeks prior to mating to cover at least two complete oestrous cycles, the variable time to conception, the duration of pregnancy and at least 4 days after delivery)

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following tissues were prepared for microscopic examination and weighed, respectively: ovaries, testes, epididymides, seminal vesicles, coagulating glands, prostate together with all gross lesions. The testes and epididymides of all adult males were weighed. The organ weights as percentage of bodyweights were also determined and recorded.

Statistics:

The significance of group differences was tested using Dunnett's 't' test and the Student's 't' test.

Reproductive indices:

Gestation index = (no. of females with live pups / no. of pregnant females) x 100

Male fertility index = (no. of males impregnating females / no. of males exposed to females) x 100

Female fertility index = (no. of pregnant females / no. of paired females) x 100

Fecundity index = (no. of pregnant females / no. of mated females) x 100

Mean no. of corpora lutea = total no. of corpora lutea / no. of dams used for corpora lutea count

Mean no. of implantations = total no. of implantation / no. of dams used for implantation count

Implantations = (total no. of implantations / total no. of corpora lutea) x 100

Parturition = (no. of parturitions / no. of pregnancies) x 100

Pre-terminal deaths of pregnant dams = (no. of pregnant animals dead / total no. of pregnant animals) x 100

Live pups born = (no. of lives pups born / no. of implantations) x 100

Pre-implantation loss = (no. of corpora lutea – no. of implantations / no. of corpora lutea) x 100

Post implantation loss = (no. of implantations – no. of live pups / no. of implantations) x 100

Offspring viability indices:

Mean litter size index = no. of pups on day 0 / no. of females littered

Mean viable litter size = no. of viable pups on day 0 / no. of females littered

Live birth index = (no. of pups alive on day at first observation / no. of pups born at first observation) x 100

24 hour survival index = (no. of viable pups on lactation day 1 / no. of viable pups born) x 100

4th day survival index = (no. of pups viable pups on lactation day 4 / no. of viable pups born) x 100

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:

not examined

Description (incidence and severity):

Test substance intake: not applicable

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
There were no pre-terminal deaths at any of the doses tested and no clinical signs were observed. The partial cannibalism of pups was observed in three females in control and three females of the high dose group and all pups cannibalism was observed in one female of the mid dose group.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Body weights and food consumption were unaffected by treatment with the test material at all dose levels.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
There were no treatment related effects observed on the mean number of days of pre-coital interval between control and the treated groups and there were no treatment related effects observed in the length of the pregnancy between control and treated groups. The mean number of male and female pups and the mean number of pups for combined sex during different intervals of lactation period at all tested doses were unaffected by the treatment when compared to control. Fertility indices of sires and dams, fedundity index, percentage of implantations, gestation index, parturition percentage, percentage of pre-implantation loss, post-implantation loss and live pups born were unaffected by the treatment at all doses tested when compared to control. Incidentally, there were higher numbers of live pups born at the mid and high doses due to the lower percentage of post implantation loss. The significantly higher percentage of pre-implantation loss resulted in lower percentage of implantations at the low and mid doses. These findings were considered incidental as there were no changes observed at the high dose.

ORGAN WEIGHTS (PARENTAL ANIMALS)
There were no significant differences in organ weights and organ weight ratios in the males.

GROSS PATHOLOGY (PARENTAL ANIMALS)
There were no treatment related gross changes in males and females. the few incidences of lesions observed were randomly distributed among the various groups and were hence considered incidental.

HISTOPATHOLOGY (PARENTAL ANIMALS)
No lesions were observed in the ovaries, testes and epididymides in the control and high dose animals examined for histopathology.

Effect levels (P0)

Dose descriptor:

NOEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No treatment related effects were observed at this dose level in any of the parameters evaluated

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:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Details on results (F1)

VIABILITY (OFFSPRING)
The number of live litters, mean litter size index, sex ratio at birth, number of pups dead at first observation, number of pups dead/cannibalised on day 1, up to day 4, the number of pups alive on day 0 and 1, live birth index and 24 hour survival index were unaffected by the treatment at all doses tested when compared to control. Any differences that occurred were a result of cannibalism and were considered incidental and not related to the treatment.

BODY WEIGHT (OFFSPRING)
the mean weight of males and female pups and the mean weight of pups for combined sex during different intervals of lactation period were unaffected by the treatment when compared to control.

GROSS PATHOLOGY (OFFSPRING)
There was a single incidence of a rudimentary tail in one high dose male pup. This was considered an incidental change and thus there were no treatment related gross changes observed in the pups.

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

It is concluded that the oral administration of the test material to rats at the concentration of 1000 mg/kg, had no effects on general health, body weights and food consumption. There were no treatment-related signs and no pre-terminal deaths. No treatment-related effects were observed on gestation and lactation body weights and food consumption, number and weight of pups, viability of pups, fertility indices of parental animals, terminal body weights, organ weights and their ratios in males. There were also no treatment-related gross and histopathological changes. Under the conditions of the study, the No Observed Effect Level was therefore determined to be 1000 mg/kg bw.

Executive summary:

The reproductive and developmental toxicity of the test material was investigated in accordance with standardised guidelines OECD 421 and EPA OPPTS 870.3550. During the study rats were dosed test material by gavage, at daily concentrations of 0 (vehicle control), 100, 400 and 1000 mg/kg bw. Following two weeks of treatment, animals were mated and treatment continued for another two weeks (minimum). Animals were observed for mortality and clinical signs during the treatment period; body weights and food consumption were also recorded. At the end of the treatment period animals were sacrificed and subjected to gross and histopathological examination. Under the conditions of the study, it was concluded that the oral administration of the test material to rats at the concentration of 1000 mg/kg, had no effects on general health, body weights and food consumption. There were no treatment-related signs and no pre-terminal deaths. No treatment-related effects were observed on gestation and lactation body weights and food consumption, number and weight of pups, viability of pups, fertility indices of parental animals, terminal body weights, organ weights and their ratios in males. There were also no treatment-related gross and histopathological changes. In consideration of the findings, the No Observed Effect Level was determined to be 1000 mg/kg bw.