reaction products of 2-Propenoic acid (2 moles) and Neopentylglycol hydroxypivalate (1 mole)

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 June 2017 - 15 August 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

29 July 2016

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

reaction products of 2-Propenoic acid (2 moles) and Neopentylglycol hydroxypivalate (1 mole)

EC Number:

821-997-6

Cas Number:

2136366-99-7

Molecular formula:

C11H16O6 to C21H32O8

IUPAC Name:

reaction products of 2-Propenoic acid (2 moles) and Neopentylglycol hydroxypivalate (1 mole)

Test material form:

liquid

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: breeder: Janvier, Le Genest-Saint-Isle, France
- Age: at the beginning of the treatment period, the males were approximately 10 weeks old and the females were approximately 11 weeks old
- Mean body weight: at the beginning of the treatment period, the males had a mean body weight of 425 g (range: 392 g to 449 g) and the females had a mean body weight of 262 g (range: 226 g to 296 g).
- Housing: F0 animals were individually housed, except during mating (males + females) and lactation (females + pups), in polycarbonate cages (Tecniplast 2154, 48 x 26.5 x 21 cm, 940 cm2) with stainless steel lids and containing autoclaved sawdust (SICSA, Alfortville, France). Individual housing of F0 animals was chosen as group housing could adversely affect gestation and lactation, and to avoid aggressive behavior between males around mating.
Toward the end of gestation and during lactation, autoclaved wood shavings (SICSA, Alfortville, France) were provided to females and their litter as nesting material.
- Diet: SSNIFF R/M-H pelleted diet (free access)
- Water: tap water filtered with a 0.22 µm filter (free access)
- Acclimation period: the males were acclimated to the study conditions for 7 days before treatment and the females were acclimated to the study conditions for 5 days before the beginning of estrous cycle monitoring during the pre-treatment period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 50 ± 20%
- Air changes (per hr): approximately 8 to 15 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12 h/12 h.

IN-LIFE DATES: 20 June 2017 to 15 August 2017.

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

The oral route was selected since it is a route of administration which is recommended by the Regulatory Authorities for this type of study. The dose formulations were administered by gavage, using a plastic syringe fitted with a plastic gavage tube, once a day, at approximately the same time.

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING FORMULATIONS: solution in the vehicle
- Concentration in vehicle: 20, 60 and 120 mg/mL
- Amount of vehicle: 5 mL/kg/day.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Type of method: Gas Chromatography with FID detection (GC-FID)
Test item concentrations: remained within an acceptable variation range (-3.4% to +5.4%) when compared to the nominal values.
Homogeneity: diluted analytical samples based on the marker peak prepared from 1.6 mg/mL and 240 mg/mL dose formulations in corn oil were found to be stable for 6 days when they were stored at room temperature and protected from light.

Duration of treatment / exposure:

In the males:
- 2 weeks before mating,
- during the mating period (until 1 week),
- until euthanasia (4 weeks in total) (until study Day 28),

In the females:
- 2 weeks before mating,
- during the mating period (until 1 week),
- during gestation,
- during lactation until Day 13 post-partum inclusive,
- until euthanasia for females which had no delivery.

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

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

Doses / concentrations 4

Dose / conc.:

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

No. of animals per sex per dose:

10 animals per sex per dose

Control animals:

yes, concurrent vehicle

Positive control:

no (not required)

Results and discussion

Results of examinations

Details on results:

All the results are in the section 7.8.1 (combined study).

Effect levels

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Target system / organ toxicity

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Based on the experimental conditions of this study:
. the No Observed Effect Level (NOEL) for parental toxicity was considered to be 600 mg/kg/day in females based on the absence of findings at this dose level; in males, given the non-adverse findings in the kidneys, the No Observed Adverse Effect Level (NOAEL) was considered to be 600 mg/kg/day,

Executive summary:

The objective of this study was to evaluate the potential toxic effects of the test item following daily oral administration (gavage) to male and female rats from before mating, during mating and, for the females, throughout gestation until Day 13 post-partum (p.p.).

This study provides information:

. on the possible health hazards likely to arise from repeated exposure over a relative limited period of time,

. on male and female reproductive performance, such as gonadal function, mating behavior, conception, development of the conceptus and parturition.

 

Methods

Three groups of ten male and ten female Sprague-Dawley rats received the test item.

The test item was administered daily by the oral route (gavage) at dose levels of 100, 300 or 600 mg/kg/day, under a constant dosage volume of 5 mL/kg/day.

Males were treated for an overall period of 4 weeks: two weeks before mating, during the mating period (up to one week) until the day before sacrifice. Females were treated for an overall period of 7 to 8 weeks: two weeks before mating, throughout mating (up to one week) and gestation (3 weeks) until Day 13post-partum(p.p.) inclusive. Another group of 10 males and 10 females received the vehicle only (corn oil) under the same experimental conditions and acted as a control group.

The actual test item concentrationsin the dose formulations were determined in Weeks 1 and 6 using a validated GC-FID analytical method.

Animals were checked daily for clinical signs and mortality. Body weights and food consumption were recorded weekly until mating and then at designated intervals throughout gestation and lactation. Estrous cycle stage was determined each morning from two weeks before mating until the females had mated and on Day 14p.p.before euthanasia.

The animals were paired for mating after two weeks of treatment and the dams were allowed to litter and rear their progeny until Day 13p.p. The total litter sizes and numbers of pups of each sex were recorded after birth. The size of each litter was adjusted on Day 4p.p.by culling extra pups to obtain as nearly as possible four males and four females per litter. Pups not selected on Day 4p.p.were sacrificed and discarded without further examination.

The pups were observed daily for clinical signs or abnormal behaviour and were weighed on Days 1, 4, 8 and 13p.p.The physical development of pups was assessed by measuring the anogenital distance on Day 1p.p.and by counting the number of nipples and areolae in male pups on Day 12p.p.

Hematology and blood biochemistry investigations were performed on at least five males and females in each group at scheduled sacrifice. Thyroid hormones (TSH and T4) were determined in males at sacrifice and in pups sacrificed on Day 13p.p.

Males were sacrificed after completion of the mating period. Dams were sacrificed on Day 14p.p.

A full macroscopicpost-mortemexamination was performed, with a particular attention to the reproductive organs. Designated organs were weighed and selected tissue specimens were preserved. A microscopic examination was performed on selected tissues from five males and lactating females in the control and high-dose groups.

Pups were sacrificed on Day 13p.p.and submitted to a detailed external examination with a particular attention to the external genital organs.

Results

Actual concentrations of the test item in the dose formulations analyzed during the study remained within an acceptable range of variation (-3.4% to +5.4%) when compared to the nominal concentrations.

 

F0 animals:

No test item-related deaths occurred in males or females during the study.

Ptyalism was observed with a dose-related incidence at all dose levels of the test item. This sign, commonly noted when a test item is administered by gavage, was not considered as an adverse effect.

Body weight and food consumption were unaffected by the test item treatment.

The estrous cycle was not impacted by the test item treatment.

The mating, fertility and delivery data were unaffected by the test item treatment.

Hematology and blood biochemistry parameters were not impacted by the test item treatment.

Thyroid hormone analyses did not reveal any disturbance in F0 males at sacrifice.

At pathology examination, no effects on organ weights and no test item-related macroscopic or microscopic findings were observed in females. In male rats, an increase in renal tubular hyaline droplets was observed. This finding is specific to male rats and has no relevance for human risk assessment.

 

Pups:

Observations of the pups from birth to Day 13p.p.did not show any effects on mortality, viability, clinical signs, body weight evolution, sex ratio or anogenital distance. The only observation consisted of the presence of areolae in one and five pups at 300 and 600 mg/kg/day, respectively. As this finding was isolated with low incidence and was no longer observed the day after in two pups at 600 mg/kg/day, this was considered to be non-adverse.

Thyroid hormone analyses did not reveal any disturbance in pups on Day 13p.p.

 

Conclusion

The test item was administered daily by oral gavage to male and female Sprague-Dawley rats, for 2 weeks before mating, during mating, and (for females) throughout gestation and until Day 13 post-partum, at dose levels of 100, 300 or 600 mg/kg/day.

 

Based on the experimental conditions of this study:

. the No Observed Effect Level (NOEL) for parental toxicity was considered to be 600 mg/kg/day in females based on the absence of findings at this dose level; in males, given the non-adverse findings in the kidneys, the No Observed Adverse Effect Level (NOAEL) was considered to be 600 mg/kg/day,

. the No Observed Effect Level (NOEL) for reproductive performance (mating and fertility) was considered to be 600 mg/kg/day based on the absence of effects on mating or fertility at this dose level,

. the NOAEL for toxic effects on progeny was considered to be 600 mg/kg/day based on the absence of adverse findings on pups at this dose level.