Reaction mass of 1,3-dioxane-5,5-dimethanol and 1,3-Propanediol, 2-(hydroxymethyl)-2-[(methoxymethoxy)methyl]-

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

This is a preliminary study to assist with setting dose levels for an Extended One-generation Reproductive Toxicity Study (OECD443)

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

06 Jul 2021 to 23 December 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

This study was conducted in a facility which operates in accordance with GLP. However no claim of GLP compliance was intended nor is made for this study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 443 (Extended One-Generation Reproductive Toxicity Study)

Version / remarks:

Dose range finding study (not under GLP) performed in order to select doses for the Main EOGRT OECD 443 study.

Deviations:

no

Principles of method if other than guideline:

The purpose of this study was to assess the general systemic toxic potential in rats, including reproductive/developmental effects, with administration of the test item by oral gavage in order to act as a preliminary study to assist with setting dose levels for an Extended One-generation Reproductive Toxicity Study (OECD443).

GLP compliance:

no

Remarks:

This DRF study was not performed under GLP, but conducted at in a facility which operates in accordance with GLP.

Limit test:

no

Justification for study design:

The purpose of this study was to assess the general systemic toxic potential in rats, including reproductive/developmental effects, with administration of the test item by oral gavage administration in order to act as a preliminary study to assist with setting dose levels for an Extended One-generation Reproductive Toxicity Study (OECD443).

Test material

Test material information

Constituent 1

Reference substance name:

1,3-dioxane-5,5-dimethanol

EC Number:

228-329-7

EC Name:

1,3-dioxane-5,5-dimethanol

Cas Number:

6228-25-7

Molecular formula:

C6H12O4

IUPAC Name:

1,3-dioxane-5,5-diyldimethanol

Constituent 2

Reference substance name:

2-(hydroximethyl)-2-(methoxymethoxy)methyl)-1,3- Propanediol

Cas Number:

68658-38-8

Molecular formula:

C7H16O5

IUPAC Name:

2-(hydroximethyl)-2-(methoxymethoxy)methyl)-1,3- Propanediol

impurity 1

Reference substance name:

Pentaerythritol

EC Number:

204-104-9

EC Name:

Pentaerythritol

Cas Number:

115-77-5

Molecular formula:

C5H12O4

IUPAC Name:

2,2-Bis(hydroxymethyl)-1,3- propanediol

impurity 2

Reference substance name:

2-(hydroxymethyl)-2-(methoxymethyl)propane-1,3-diol

Cas Number:

55502-62-0

Molecular formula:

C6H14O4

IUPAC Name:

2-(hydroxymethyl)-2-(methoxymethyl)propane-1,3-diol

impurity 3

Reference substance name:

5-[(hydroxymethoxy)methyl]-1,3-dioxane-5-methanol

EC Number:

301-313-8

EC Name:

5-[(hydroxymethoxy)methyl]-1,3-dioxane-5-methanol

Cas Number:

93983-24-5

Molecular formula:

C7H14O5

IUPAC Name:

{5-[(hydroxymethoxy)methyl]-1,3-dioxan-5-yl}methanol

impurity 4

Reference substance name:

Other impurities

IUPAC Name:

Other impurities

Test material form:

liquid: viscous

Details on test material:

Batch no.: Lot No. 90000029663
List no.: 911-819-6
Storage: room temperature (20 ± 5 °C); store in sealed containers at ambient temperature, in a dry area protected from direct sunlight and the elements
Expiry date: 26. Aug. 2022
Stability: stable under storage conditions
Appearance: amber, turbid, viscous liquid
Homogeneity: homogeneous
Vapour pressure: 0.02 Pa at 25 °C (MPBPWIN v1.43 Modified Grain method) Vapour study protocol approved by EPA and study to be run
Solubility in solvents H2O: > 1000 g/L (OECD 105; submitted to EPA on November 21, 2018 and accepted); EtOH: not stated; acetone: not stated; CH3CN: not stated; DMSO: not stated

Specific details on test material used for the study:

- Name of test material (as cited in study report): Polyol PX; alternative name: Reaction mass of 1, 3-Propanediol, 2-(Hydroxymethyl)-2-[(Methoxymethoxy)methyl]-and 1, 3-Dioxcine 5, 5-Dimethanol
- Source: Sponsor
- Physical state: Solid
- Purity: 71.2% (dose calculations were corrected for purity; correction factor 1.40)
- Lot/batch No.: 90000029663
- Expiration date of the lot/batch: 26 August 2022
- Stability and homogeneity: Homogeneity and stability of the test material in the vehicle was determined as part of this programme of work in Labcorp Study No. 8459847. Storage was confirmed at ambient temperature (15 to 25ºC) for up to 1 day and following refrigeration (2 to 8°C) for up to 15 days following fresh preparation in purified water for the range from 2 mg/mL and 200 mg/mL.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Remarks:

Crl:CD(SD)

Details on species / strain selection:

The rat was chosen as the test species because of the requirement for a rodent species by regulatory agencies. The Sprague Dawley [Crl:CD(SD)] strain was used because of the historical control data available at this laboratory.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: (P) 10-12 wks; (F1) nominally 28 days
- Weight at study initiation: (P) Males: 340-428 g; Females: 230-275 g
- Housing:
Cages comprised of a polycarbonate body with a stainless steel mesh lid; changed at appropriate intervals. Solid (polycarbonate) bottom cages were used throughout the study except during pairing.
Grid bottomed cages were used during pairing. These were suspended above absorbent paper which was changed daily.
Pre-pairing phase up to four animals of one sex per cage
Pairing one male and one female per cage
Males after mating up to four animals per cage
Gestation one female per cage
Lactation one female + litter per cage
Selected F1 generation (from Day 21 of age) up to four animals of one sex per cage

- Diet: ad libitum. SDS VRF1 Certified pelleted diet. The diet contained no added antibiotic or other chemotherapeutic or prophylactic agent.
- Water: ad libitum. Potable water from the public supply via polycarbonate bottles with sipper tubes. Bottles were changed at appropriate intervals.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 40-70
-Air changes: Filtered fresh air which was passed to atmosphere and not recirculated
- Photoperiod 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 14 Jul 2021 to19 Oct 2021

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The test item was heated to 40°C in a water bath and the required amounts weighed. Approximately 50% of the required vehicle was added to the test item and magnetically stirred until all the test item was uniformly mixed. Further amounts of vehicle were gradually added and mixed to produce the required concentration. The formulation was finally mixed using a high-shear homogenizer and transferred, via syringe, into final containers. Start and finish times recorded in the raw data. The dosing formulations were prepared weekly, stored in a refrigerator set to maintain 2 to 8 °C.

The dosing volume was 5 mL/kg.

A correction factor of 1.40 was used to calculate the amount of test item required during formulation.

Details on mating procedure:

One female to one male within the same treatment group until evidence of mating.
Evidence of mating was ejected copulation lugs and sperm in the vaginal smear.
Day 0 of gestation was set to the day positive sign of mating was detected.
Maximum duration of pairing was 2 weeks.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

No formulation analysis was performed on this study

Duration of treatment / exposure:

F0: For two weeks before pairing until termination after litters were weaned.
F1: From weaning on Day 21 until attainment of sexual maturity (approximately Week 7 of age).

Frequency of treatment:

Once daily at approximately the same time each day. Animals were not dosed if parturition was in progress at the scheduled time of administration.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Negative control

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Low dose

Doses / concentrations 3

Dose / conc.:

600 mg/kg bw/day (nominal)

Remarks:

Intermediate dose

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

High dose

No. of animals per sex per dose:

F0 generation: 8 males and 8 females per dose group
F1 generation: 10 males and 10 females per dose group

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale:
Dose levels of 0, 300, 600 and 1000 mg/kg/day were selected in conjunction based on the findings in an acute single dose study in the rat (Report no. CTL/T/1741), a repeat dose toxicity study in Wistar rats (Study No. 528617) and a developmental toxicity study in the Han Wistar rat (Study No. 497207).
In the Acute study the LD50 was in excess of 5000 mg/kg bw.
In the 90-day repeat dose study the no observed adverse effect level was 1000 mg/kg/day.
In the Developmental toxicity study the no observed adverse effect level was 1000mg/kg/day.

The doses selected were of a reduced dose interval in order to determine a suitable high dose for the main OECD443 study.

Positive control:

Not required

Examinations

Parental animals: Observations and examinations:

CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily
Animals were inspected visually at least twice daily for evidence of ill-health or reaction to treatment.
-During littering: Observed approximately 24 hours after birth (Day 1 of age) and then daily for evidence of ill-health or reaction to treatment.


PHYSICAL EXAMINATION: Yes
-Time schedule: A detailed physical examination was performed on each animal to monitor general health
All F0 and selected F1 generation animals: Once each week
In addition: F0 females: Days 0, 5, 12, 18 and 20 after mating and Days 1, 7, 14 and 21 of lactation.

DETAILED CLINICAL OBSERVATIONS: Yes
F0 males: Week 1 - Daily, Weeks 2 to 4 - twice weekly (middle and end of the week), Week 5 onward - once each week
F0 females: Week 1 - Daily, Weeks 2 - twice (middle and end of the week), After mating - Days 0, 7, 14 and 20, Lactation - Days 1, 7, 14 and 20
- Time schedule: Detailed observations were recorded at the following times in relation to dose administration:
*Pre-dose observation.
*One to two hours after completion of dosing.
*As late as possible in the working day.


BODY WEIGHT: Yes
- Time schedule for examinations:
F0 males: Twice weekly, Before necropsy.
F0 females: Twice weekly until mating detected. Days 0, 3, 7, 10, 14, 17 and 20 after mating.
Days 1, 4, 7, 11, 14, 18 and 21 of lactation. Before necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: yes
- Time schedule for examinations: The weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded as follows:
F0 males: Weekly, until paired for mating.
F0 females: Weekly until mating detected. Days 0-2, 3-6, 7-9, 10-13, 14-16 and 17-19 after mating.
Days 1-3, 4-6, 7-10, 11-13, 14-17 and 18-20 of lactation.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No (ad libitum)

DURATION OF GESTATION: Time that elapsed between mating and commencement of parturition.

PATURITION OBSERVATION: From Day 20 after mating animals were checked three times daily for evidence of parturition. The progress and completion of parturition was monitored; numbers of live and dead offspring were recorded and any difficulties observed were noted.

Oestrous cyclicity (parental animals):

Dry smears were taken for 15 days before pairing, using cotton swabs.
Wet smears were taken after pairing until evidence of mating confirmed, using pipette lavage.

Litter observations:

PARAMETERS EXAMINED
The clinical condition, litter size and survival, sex ratio, body weight and macropathology of all F1 offspring were assessed. After weaning, selected F1 generation animals were assessed for clinical condition, body weight, food consumption, sexual maturation and macropathology investigations.

CLINICAL OBSERVATIONS: Yes
- Time schedule: at least twice daily
Animals were inspected visually at least twice daily for evidence of ill-health or reaction to treatment.

PHYSICAL EXAMINATION: Yes
-Time schedule: A detailed physical examination was performed on selected F1 generation animals to monitor general health once each week.

DETAILED CLINICAL OBSERVATIONS: Yes
F1 selected animals: From Day 21 of age up to Week 2 of the F1 generation-daily, From Week 2 - twice weekly
- Time schedule: Detailed observations were recorded at the following times in relation to dose administration:
*Pre-dose observation.
*One to two hours after completion of dosing.
*As late as possible in the working day.

BODY WEIGHT: Yes
- Time schedule for examinations:
- Individual offspring body weight: Recorded on Days 1, 4 (before culling), 7, 14, 17 and 21 of age.
-F1 selected animals - Day 21 and Day 25 of age and then twice weekly from nominal four weeks of age to termination at approximately seven weeks of age.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
-Time schedule for examinations: F1 selected animals twice weekly from nominal four weeks of age to termination.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No (ad libitum)

LITTER SIZE
-Daily records were maintained of mortality and consequent changes in litter size from Days 1-21 of age. On Day 4 of age, litters containing more than ten offspring were reduced to ten by random culling, leaving, whenever possible, five male and five female offspring in each litter.
- Grossly normal offspring were discarded. Culled offspring with clinical signs on Day 4 of age were individually identified for subsequent macroscopic examination.

SEX RATIO OF EACH LITTER
- Recorded on Days 1, 4 (before and after culling) and on Day 21 of age.

SEXUAL MATURATION - F1 Generation
- Males: Sexual maturation was assessed by daily examination from Day 38 of age until balano-preputial separation occurred. Body weight was recorded on the day of completion of separation.
-Females: Sexual maturation was assessed by daily examination from Day 28 of age until vaginal opening occurred. Body weight was recorded on the day of vaginal opening.

Postmortem examinations (parental animals):

SACRIFICE
- F0 males: After successful littering by females.
- F0 females: Day 21 of lactation.

All adult animals were subject to a detailed necropsy. After a review of the history of each animal, a full macroscopic examination of the tissues was performed. All external features and orifices were examined visually. Any abnormality in the appearance or size of any organ and tissue (external and cut surface) was recorded and the required tissue samples preserved in appropriate fixative.

For each uterine horn the number of implantation sites were recorded.

Postmortem examinations (offspring):

SACRIFICE
- Premature deaths before weaning (excluding culled offspring): Subject to complete macroscopic examination with assessment of stomach for presence of milk, where possible. Abnormal tissues retained in an appropriate fixative.
- Culled offspring on Day 4 of age: Culled offspring with clinical signs on Day 4 of age were subject to complete macroscopic examination with assessment of stomach for presence of milk, where possible. Abnormal tissues retained in an appropriate fixative. Culled offspring with no clinical signs on Day 4 of age were killed and discarded without necropsy examination.
-Unselected offspring at scheduled kill on Day 21 of age and selected spares: Subject to complete macroscopic examination. Abnormal tissues retained in an appropriate fixative.
-F1 adult offspring: Scheduled kill- approximately week 7 of age (after completion of sexual maturation)

Statistics:

Statistical analyses were performed on the majority of data presented and results of these
tests, whether significant or non-significant, are presented on the relevant tables. For
pre-coital interval, mating performance and fertility and gestation index the similarity of the
data was such that analyses were not considered to be necessary.

All statistical analyses were carried out separately for males and females. Data relating to food consumption were analyzed on a cage basis. For all other adult parameters, the analyses were carried out using the individual animal as the basic experimental unit. For litter/fetal findings the litter was taken as the treated unit and the basis for statistical analysis and biological significance was assessed with relevance to the severity of the anomaly and the incidence of the finding within the background control population.

The following data types were analyzed at each timepoint separately:
- Body weight, using absolute weights and gains over appropriate study periods
- Food consumption, over appropriate study periods
- Estrous cycles
- Gestation length
- Litter (implantations, litter size, sex ratio - percentage male, post implantation survival index, live birth index and viability index), for before cull study periods
- Sexual maturation, age and body weight at completion

The following pairwise comparisons were performed: Group 1 vs 2, 3 and 4
Significant differences between the groups compared were expressed at the 5% (p<0.05) or 1% (p<0.01) level.

Parametric/Non-Parametric tests include:
- Bartlett's test, Willams' test, t-tests, Dunnett's tests, Shirley's test Kruskal-Wallis' tests, Fisher's exact test, Wilcoxon rank sum tests, Cochran-Armitage tests, Chi-square tests, one- and two-tailed linear-by-linear tests.

Reproductive indices:

Percentage mating (%) = (Number of animals mating/Animals paired) x 100

Conception rate (%) = (Number of animals achieving pregnancy/Animals mated) x 100

Fertility index (%) = (Number of animals achieving pregnancy/Animals paired) x 100

Gestation index (%) = (Number of live litters born/Animals paired) x 100

Post implantation survival index (%) = (Total number of offspring born)/(Total number of uterine implantation sites) x 100

Live birth index (%) = (Number of offspring on Day 1 after littering)/(Total number of offspring born) x 100

Offspring viability indices:

Viability index (%) = (Number of live offspring on Day 4 before culling)/(Number of live offspring on Day 1 after littering) x 100

Lactation index (%) = (Number of live offspring on Day 21 after littering)/(Number of live offspring on Day 4 (after culling)) x 100

Percentage males = (Number of males in litter/Total number of offspring in litter) x 100

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Initially over Days 1 to 5 of treatment males at 1000 mg/kg/day showed high body weight gain when compared with Controls (p<0.05) and during Days 15 to 19 of treatment males at 600 or 1000 mg/kg/day showed low weight gain (p<0.05). However overall body weight and body weight gain for males during the treatment period was unaffected by administration of Polyol PX at dose levels up to and including 1000 mg/kg/day.

During the two week treatment period before pairing females receiving Polyol PX showed low mean body weight gain when compared with Controls with statistical significance attained at 600 and 1000 mg/kg/day (p<0.05) but with no dose response.

During gestation bodyweight gain for females receiving 1000 mg/kg/day was slightly low from Day 7 of gestation and overall, the body weight gain from GD0-20 was statistically significantly low when compared with Controls (p<0.05); gestation body weight gain at 300 or 600 mg/kg/day showed no adverse effect of treatment.

Overall, the bodyweight gain for females during lactation was higher for females receiving Polyol PX with statistical significance attained at 300 and 1000 mg/kg/day (p<0.01); there was no evidence for a dose response.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

Overall, food consumption for both males and females during the two-week treatment period before pairing and females during gestation showed no adverse effects at dose levels up to and including 1000 mg/kg/day.

During Days 11-13 of lactation food consumption for females at 1000 mg/kg/day was slightly low when compared with Controls (p<0.05); food consumption at 100 or 300 mg/kg/day was essentially similar to Controls.

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

Endocrine 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 examined

Histopathological findings: neoplastic:

not examined

Other effects:

not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

not specified

Description (incidence and severity):

The gestation length of treated females was within the expected range of 22 to 23 days, however females receiving Polyol PX tended to show a longer gestation period when compared with Controls, with the shift in distribution showing statistical significance at 1000 mg/kg/day (p<0.05). The gestation index was unaffected by treatment.

Details on results (P0)

Pre-coital interval, mating performance and fertility were unaffected by treatment.

Effect levels (P0)

Key result

Dose descriptor:

other:

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other:

Target system / organ toxicity (P0)

Key result

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

On Day 1 of age offspring bodyweight at 1000 mg/kg/day was high at approximately 111%
of Controls for male offspring (p<0.01) and 107 % of Controls for female offspring (not
statistically significant). Subsequent body weight gain up to weaning was unaffected by
administration of Polyol PX at dose levels up to and including 1000 mg/kg/day.

Body weight and body weight gain for both selected males and females from Day 21 to Day 25 of age were unaffected by treatment.

Body weight on Day 1 of the F1 generation at nominal PND28±2 and subsequent gain up to Day 4 for selected males was low at all dose levels (p<0.05 and p<0.01 respectively); there was no evidence of a dose response. Subsequent gain was similar to Controls and mean body weight prior to termination was essentially similar across the groups.

Body weight on Days 1 and 4 of the F1 generation (PND28±2) was low for selected females at 600 mg/kg/day (p<0.01); however, this was not apparent at either 300 or 1000 mg/kg/day. Overall body weight gain for females receiving 600 or 1000 mg/kg/day was high when compared with Controls (p<0.05 and p<0.01, respectively).

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

Sexual maturation:

no effects observed

Anogenital distance (AGD):

not examined

Nipple retention in male pups:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

not examined

Other effects:

not specified

Description (incidence and severity):

At 1000 mg/kg/day the mean number of implantations and litter size on PND1 and 4 were slightly low at approximately 84 to 89% of Controls but these differences did not attain statistical significance. Litter size at 100 or 300 mg/kg/day was unaffected by treatment.

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Details on results (F1)

Offspring survival and sex ratio at dose levels up to and including 1000 mg/kg/day showed no adverse effects of treatment.

Effect levels (F1)

Key result

Dose descriptor:

other:

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other:

Target system / organ toxicity (F1)

Key result

Critical effects observed:

no

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Applicant's summary and conclusion

Conclusions:

Based on the results of this preliminary study, and in the absence of any overt toxicity, there is nothing to preclude the use of 1000 mg/kg/day as the high dose in the subsequent OECD 443 study.

Executive summary:

The purpose of this study was to assess the general systemic toxic potential in rats, including reproductive/developmental effects, with administration of the test item by oral gavage administration in order to act as a preliminary study to assist with setting dose levels for an Extended One-generation Reproductive Toxicity Study (OECD 443).

In the F0 generation, three groups of eight male and eight female Sprague Dawley rats received Polyol PX at dose levels of 300, 600 or 1000 mg/kg/day at a volume dose of 5 mL/kg/day. Males were treated for two weeks before pairing, up to necropsy after litters were weaned. Females were treated for two weeks before pairing, throughout pairing up to necropsy on Day 20 of lactation. In the F1 generation, ten males and ten females were treated from weaning to their scheduled termination (after attainment of sexual maturation) at the same dose levels and volume-dose as the F0 generation. Similarly constituted Control groups received the vehicle, purified water, at the same volume dose and throughout the same relevant period. 

During the study, clinical condition, body weight, food consumption, mating performance, gestation length, parturition observations and macropathology investigations were undertaken on F0 adults. The clinical condition, litter size and survival, sex ratio, body weight and macropathology of all F1 offspring were assessed. After weaning, selected F1 generation animals were assessed for clinical condition, body weight, food consumption, sexual maturation and macropathology investigations.

Oral gavage administration of Polyol PX at dose levels of 300, 600 and 1000 mg/kg/day was generally well tolerated with no adverse effects on clinical condition, food consumption, reproductive performance, offspring survival, offspring development, sexual maturation or macropathology. Minor effects were limited to the high dose group which exhibited a slightly longer gestation length which may be associated with the slightly lower number of implantations and smaller litter size. Based on the results of this preliminary study, and in the absence of any overt toxicity, there is nothing to preclude the use of 1000 mg/kg/day as the high dose in the subsequent
OECD 443 study.