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Administrative data

Key value for chemical safety assessment

Effects on fertility

Effect on fertility: via oral route
Endpoint conclusion:
no study available
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

No evidence of an effect on the reproductive organs was seen in a 90-day study at dose levels of up to and including 1000 mg/kg bw/d. Similarly, no evidence of any effects on the developing fetus was seen in a prenatal developmental toxicity study at the limit dose level of 1000 mg/kg bw/d. The absence of histopathological changes in reproductive organs in the repeated dose toxicity study indicate that the substance will not have any adverse effects on fertility. Based on these considerations, no further testing is required for fertility assessment.


Short description of key information:
No evidence of any potential for reproductive toxicity was seen in a 90-day study at dose levels of up to and including 1000 mg/kg bw/d. Similarly, no evidence of any effects on the developing fetus was seen in a prenatal developmental toxicity study at the limit dose level of 1000 mg/kg bw/d. A waiver is therefore proposed for the extended one generation reproductive toxicity study based on the absence of effects seen in the repeat dose study and the developmental toxicity study.

Justification for selection of Effect on fertility via oral route:
No study available - a waiver is proposed.

Effects on developmental toxicity

Description of key information
An oral prenatal developmental toxicity is available for Polyol PX; no effects were observed up to and including the highest dose tested therefore the NOELs were considered to be 1000 mg/kg bw/d for both maternal toxicity and embryofetal toxicity
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23 September 2015 to 01 February 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Modern GLP and guideline compliant study
Qualifier:
according to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Han Wistar (Crl:WI(Han)) - time mated females
- Source: Charles River UK Limited, Margate, UK
- Age at study initiation: 9 weeks old at time of mating
- Weight at study initiation: 177-267 g (170 -250 g on arrival)
- Housing: 2 per cage in suspended polycarbonate/polypropylene cages with stainless steel grid tops and solid bottoms. Bedding was sterilised white wood shavings.
- Diet (e.g. ad libitum): SDS VRF-1 breeder diet, ad libitum
- Water (e.g. ad libitum): water from the public supply, ad libitum
- Acclimation period: 3-5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23°C
- Humidity (%): 38-62%
- Air changes (per hr): Ten or more with 100% fresh air
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle

IN-LIFE DATES: From: 25 September 2015 To: 14 October 2015
Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
The required amount of test item was accurately weighed and transferred into a pre-labelled container according to instructions from the formulation computerised system. The appropriate amount of control item was then added to the container and the formulation was magnetically stirred until visibly homogenous. The dosing formulations were prepared weekly, stored in a refrigerator set to maintain 4°C, and dispensed daily. The dosing formulations were removed from the refrigerator and were stirred for at least 30 minutes before dosing and continuously during dosing.

The dosing volume was 10 mL/kg.

VEHICLE
- Concentration in vehicle: 0, 10, 30, 100 mg/mL)

Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Concentration analyses were performed from all dosing formulations during Week 1 and Week 2 by HPLC (reverse phase) with ELSD detection using a validated analytical procedure. Homogeneity was determined in all test substance groups during Week 1 and Week 2.

Duplicate top, middle and bottom samples (middle only for control) of 1 mL for each sampling time point were collected and sent to the analytical laboratory for analysis. Triplicate sets of top, middle and bottom samples (middle only for control) of the same volume were also collected and retained as backup samples. Concentration results were considered acceptable if mean sample concentration results were within 10% of theoretical concentration. Homogeneity results were considered acceptable if the relative standard deviation of the mean value at each sampling location was ≤10% for each group.

Stability analyses were performed during a previous study at the test facility and confirmed that the test substance is stable in the vehicle when prepared and stored under the same conditions at concentrations bracketing those used in the present study for at least 8 days at 2-8°C, in the dark.
Details on mating procedure:
Time-mated female rats were delivered to the test facility
Duration of treatment / exposure:
Days 6-19 of gestation
Frequency of treatment:
Daily
Duration of test:
Days 6-19 of gestation (scheduled euthanasia on Day 20)
No. of animals per sex per dose:
24 females/dose
Control animals:
yes, concurrent vehicle
Details on study design:
The dose levels were chosen by the Sponsor following review of data from a 14 day study using Polyol PX (Charles River Study Number 528596) and in attempt to produce graded responses to the test item. On Study 528596, there were no treatment related clinical observations or any body weight or food consumption effects at dose levels of up to 1000 mg/kg bw/d.
Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily; once in the morning and once towards the end of the working day for general health/mortality and moribundity. Animals were also observed prior to dosing and regularly throughout the day for reaction to treatment on each day of dosing.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once pretreatment, and daily from Day 6 of gestation until study termination

BODY WEIGHT: Yes
- Time schedule for examinations: on Day 4 gestation and daily from Days 6 to 20 of gestation

FOOD CONSUMPTION: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes

WATER CONSUMPTION): Yes
- Time schedule for examinations: daily visual inspection of water bottles

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 20
- Organs examined: External examination, contents of cranial, thoracic and abdominal cavities examined macroscopically (samples of abnormal tissues fixed in neutral buffered 10% formalin). The reproductive tract was examined separately (see below).

OTHER:
Ovaries and uterine content:
Gravis uterus weight was recorded and the fetuses were removed. The ovaries and uterus were examined for number and distribution of corpora lutea, implantation sites, placentae (size, colour or shape - only abnormalities were recorded), live and dead fetuses, early and late resorptions.
Fetal examinations:
Each fetus was examined for external abnormalities. Late resorptions and dead fetuses were examined for external abnormalities to the extent possible. Each implant was classified as being live, or a dead fetus (dead full term fetus that showed no signs of maceration), or a late embryonic death (macerated tissue identifiable as an embryofetus, with recognizable external features such as tail, limbs, mouth and nares present; attached to distinct identifiable placentae), or an early embryonic death (discrete, formless, discoloured tissue mass attached to the internal uterine wall; may have been of varying size). The body weight of each fetus was recorded and fetuses were individually identified within each litter.

Visceral examination and sex: Half of the viable fetuses from each uterus were fixed in methylated ethyl alcohol, examined internally for sex and eviscerated following fixation; the viscera were not examined from fetuses prior to disposal. The remaining half of viable fetuses from each uterus were fixed in Bouins’ fluid. The fetuses fixed in Bouins’ fluid were examined for soft tissue abnormalities and sex by a freehand sectioning technique derived from Wilson.

Skeletal examination: The eviscerated carcasses were then macerated in potassium hydroxide, the skeletons stained with Alizarin Red S, and then the fetuses cleared with aqueous glycerol solutions. These preparations were examined for the presence of skeletal abnormalities and for the extent of ossification.
Statistics:
Means and standard deviations were calculated for body weight, food consumption and selected pregnancy data. Numerical data collected on scheduled occasions were analysed according to sex and occasion. Descriptive statistics number, mean and standard deviation (or %CV or SE when deemed appropriate) are reported whenever possible. Inferential statistics were performed when possible, but excluded semi-quantitative data and any group with less than 3 observations. Levene’s test was used to assess the homogeneity of group variance parametric assumption at the 5% significance level. Datasets with at least three groups were compared using an overall one-way ANOVA F-test or Kruskal-Wallis test (if parametric assumptions were not met) at the 5% significance level. The pairwise comparisons were conducted using a twosided Dunnett’s or Dunn’s test, respectively, if the overall test is significant. Datasets with two groups were compared using a two-sided t-test or Wilcoxon Rank-Sum test, respectively. All significant pairwise comparisons were reported at the 0.1, 1 and 5% significance levels.
Indices:
Not applicable.
Historical control data:
Available at the test facility.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
There were no deaths and no clinical signs of toxicity. Group mean bodyweights and group mean food consumption were similar between all groups throughout the study. There were no abnormalities detected at necropsy.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Pregnancy performance and fetal weights were similar between all groups. Slight intergroup differences were considered to be incidental as they did not follow treatment related patterns and/or were too small to be attributed to treatment with Polyol PX.
The type and distribution of major and minor fetal abnormalities and skeletal ossification parameters did not indicate any association with treatment. At 300 or 1000 mg/kg/day, a slightly lower number of fetuses with 13 complete ribs and slightly increased number of fetuses with vestigial/reduced supernumerary ribs on the 1st lumbar vertebra were noted when compared with Controls. These findings are normal variation seen within litters; therefore, since the number of litters with these findings was generally similar across all groups, and there was no maternal toxicity or other correlating fetal findings to indicate that the differences were treatment related effects, the findings were considered to be incidental and unrelated to treatment with Polyol PX. Slight intergroup differences in other fetal parameters did not follow treatment related patterns and were also considered to be incidental and unrelated to treatment with Polyol PX.
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Abnormalities:
not specified
Developmental effects observed:
not specified

Dose formulation analyses: All analysed Group 2, 3 and 4 formulations from Study Weeks 1 and 2 were within the concentration acceptance criteria of ±10% of theoretical concentration, with the exception of Group 4 Week 2 samples, which were -11.6% of theoretical concentration. Back samples Group 4 Week 2 samples were therefore analysed, which were found to be within the acceptance criteria. A low relative standard deviation (≤3.2%) indicated that all formulations were homogenous. From review of data, no obvious reason could be found for the original Group 4 Week 2 results being out of acceptance; however, as the deviation was minor and backup samples were within acceptance, it is considered that animals likely received accurate formulations. The absence of Polyol PX from analysed control formulations was also confirmed.

Conclusions:
The NOAEL for maternal and developmental toxicity was considered to be 1000 mg/kg bw/d.
Executive summary:

The effects of oral administration of Polyol PX in pregnant rats during the embryo-fetal period was investigated in a study according to OECD 414. Time-mated female Han Wistar rats were administered the test substance in water at 0, 100, 300 or 1000 mg/kg bw/d by once daily by gavage. Animals were dosed over Days 6-19, inclusive, of gestation (where the day of detection of mating was Day 0 of gestation). The animals were checked regularly for clinical signs of toxicity, body weight and food consumption performance and were killed on Day 20 of gestation for examination of pregnancies and embryo-fetal development.

Dosing with Polyol PX at dose levels of up to 1000 mg/kg/ bwd was not associated with any treatment related clinical signs, body weight or food consumption effects or any macroscopic necropsy findings. Pregnancy performance and fetal weights were similar between Controls and treated groups, and the type and distribution of major and minor fetal abnormalities and variants, including skeletal ossification parameters, did not indicate any association with treatment with Polyol PX. It was, therefore, concluded from the results of this study, that the maternal and fetal no observed adverse effect levels were 1000 mg/kg bw/d.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
A modern GLP and guideline-compliant study is available for Polyol PX
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

A modern GLP and guideline compliant (OECD 414) prenatal developmental toxicity study was conducted with Polyol PX in the rat (McConnachie, 2016). Four groups of 24 females Sprague-Dawley rats were dosed orally by gavage on Days 6-19 of gestation (where Day 0 was the day of detection of mating), at dose levels of 0, 100, 300 or 1000 mg/kg bw/d in water for irrigation. At dose levels up to and including 1000 mg/kg/day were no test item-related effects on clinical observations, body weight gain, food consumption, gross pathology findings, pregnancy performance parameters, embryofetal survival or fetal weights, fetal abnormalities and variants. In conclusion, under the conditions of this study, the maternal and embryofetal no observed adverse effect levels (NOAEL) were considered to be 1000 mg/kg bw/d.


Justification for selection of Effect on developmental toxicity: via oral route:
Modern GLP and guideline compliant study in the rat

Justification for classification or non-classification

No evidence of maternal toxicity or developmental toxicity potential was observed in a prenatal developmental toxicity study conducted withe pentaerythritol, at dose levels up to and including 1000 mg/kg bw/d.

No classification is therefore required according to EC Regulation 1272/2008.