reaction mass of 2,2,3,3,5,5,6,6-octafluoro-4-(1,1,1,2,3,3,3-heptafluoropropan-2-yl)morpholine and 2,2,3,3,5,5,6,6-octafluoro-4-(heptafluoropropyl)morpholine

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:

28 January 2011

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

473-390-7

EC Name:

-

Cas Number:

1093615-61-2

Molecular formula:

C7F15NO

IUPAC Name:

2,2,3,3,5,5,6,6-octafluoro-4-(1,1,1,2,3,3,3-heptafluoropropan-2-yl)morpholine; 2,2,3,3,5,5,6,6-octafluoro-4-(1,1,2,2,3,3,3-heptafluoropropyl)morpholine

Constituent 2

Reference substance name:

MTDID 7145

IUPAC Name:

MTDID 7145

Details on test material:

- Name of test material (as cited in study report): MTDID-7145
- Physical state: Clear colorless liquid
- Analytical purity: 94%
- Lot/batch no.: 040031
- Expiration date of the lot/batch: December 2012
- Storage condition of test material: ambient/dark

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

EST ANIMALS
- Source: Charles River UK Limited, Margate, Kent, UK.
- Age at study initiation: 6 weeks
- Weight at study initiation: 180-200g (males), 150-170g (females)

The animals were initially housed 2 per cage, in polycarbonate cages, with solid bottoms andstainless steel mesh tops and measured ca 48 x 37.5 x 25 cm. A stainless steel food hopper
and polycarbonate water bottles were provided for each cage and sterilised wood shavingswere provided as bedding. Male and female cages were racked separately.A few days prior to pairing for mating, males were transferred to individual cages with astainless steel grid insert measuring ca 48 x 37.5 x 25 cm. Excreta were collected on a tray lined with absorbent paper suspended beneath each cage.
The mated females were transferred to individual solid bottomed cages measuring ca 48 x 37.5 x 25 cm. White paper tissue was supplied as nesting material from Day 20 of gestation. Females with litters retained this cage type until termination. After mating the males remained singly housed until termination.

- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 13 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.1 plus/minus 2 degrees
- Humidity (%): 55 plus/minus 15%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

other: oral gavage, suspension

Vehicle:

other: 0.5% Natrosol 250HX and 0.1% Tween 80 in Water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

- Rate of preparation of diet (frequency): Daily
- Mixing appropriate amounts with: 0.5% Natrosol 250HX and 0.1% Tween 80 in Water
- Storage temperature. 2 to 8 degrees C in dark.


VEHICLE
- Justification for use and choice of vehicle (if other than water):
0.5% Natrosol 250HX and 0.1% Tween 80 in Water
- Concentration in vehicle: To achieve 5 ml/kg vehicle doing concentrations
- Amount of vehicle (if gavage): 5 mg/kg

Details on mating procedure:

M/F ratio per cage: one male per one female
- Length of cohabitation: up to 14 days per protocol

- Proof of pregnancy:

Vaginal plug or sperm in vaginal smear

In treated females prior to mating, there was a slight reduction in group mean body weight gains and food consumption over the first two weeks of treatment, compared with Control; these reductions achieved statistical significance in animals treated at 500 and
1000 mg/kg/day. However, absolute body weights and food consumption of the females was also slightly lower than Control prior to the treatment period, and therefore this slight reduction that persisted from commencement of treatment could not be positively attributed to treatment.

Group mean body weight gains and food consumption in the males throughout the study, and for females during gestation and lactation were similar to Control.

Mating performance, fertility, duration of gestation, litter size and survival, and litter and pup weights did not indicate any obvious effect of treatment at any of the dose levels tested.

- After successful mating each pregnant female was caged (how):
The mated females were transferred to individual solid bottomed cages measuring ca 48 x37.5 x 25 cm.

- Any other deviations from standard protocol: None

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The results of the analysis of dosing formulations prepared for use on the first week of dosing (prepared on the 02 August 2010) for groups 2, 3 and 4 were outside the acceptance criteria of
± 20% (24.5 to 30%). Due to this result, a further analysis was scheduled for formulations prepared for use on the third day of dosing (04 August 2010); all results from this analysis
were ± 20 % acceptance criteria indicating acceptable accuracy of formulation. The low coefficients of variation (<6%) indicated that these formulations were homogeneous. A full review of all the data from the formulations prepared for the first day of dosing indicated that all formulations were made correctly and it is not clear why the initial results were outside the acceptance criteria. The analysis of dosing formulations for use on week 3 of dosing were within the ± 20% acceptance criteria (-10.4% to 13.5%) with low coefficient of variation, indicating acceptable accuracy of formulation.

Duration of treatment / exposure:

4 weeks male commencing 2 weeks prior to mating.
The females were dosed once daily from 2 weeks prior to mating then continued until at least Day 4 of lactation. Dosing for males and females continued until the day prior to termination.

Frequency of treatment:

once daily during the exposure phase

Details on study schedule:

-

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 500 and 1000 mg/kg/day
Basis:
analytical conc.

No. of animals per sex per dose:

10

Control animals:

yes

Details on study design:

- Dose selection rationale: Other toxicity studies

Positive control:

none

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice/day
- Cage side observations checked in table were included.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily
BODY WEIGHT: Yes
- Time schedule for examinations:Daily

Litter observations:

STANDARDISATION OF LITTERS

- Performed on day 4 postpartum: Yes


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

GROSS EXAMINATION OF DEAD PUPS:
Yes

Postmortem examinations (parental animals):

SACRIFICE

Adult animals were sacrificed by exposure to carbon dioxide followed by exsanguination.

Necropsy of Adults
All adult animals were subjected to necropsy. Necropsy consisted of an external examination, followed by macroscopic examination of the tissues and organs of the cranial, thoracic and abdominal cavities. Any gross lesions were described in terms of location, size, shape, colour, consistency, number and any other relevant characteristics. Representative samples of abnormal tissues were taken and fixed in neutral buffered 10% formalin. The following
organs were also fixed:

Ovaries uterus, cervix and vagina
Testes (weighed individually), fixed in Modified Davidson’s fluid
Epididymides (weighed individually), fixed in Modified Davidson’s fluid
Seminal vesicles and coagulating gland
Prostate gland
Pituitary gland
Skin and Mammary

The reproductive tract of all females was examined for signs of implantation with the number of implantation sites being recorded.

Postmortem examinations (offspring):

The pups were sacrificed by intra-peritoneal injection of sodium pentobarbitone.

Offspring killed or found dead were sexed, and then checked for the presence of milk in the stomach and the presence of externally visible abnormalities. Any abnormal pups were preserved in 10% formalin or methylated ethyl alcohol as appropriate for possible future examination. Externally normal decedents were discarded.

Reproductive indices:

fertitily index gestation index, Birth Index, Live Birth Index, Viability Index

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

Details on results (P0)

In treated females prior to mating, there was a slight reduction in group mean body weight gains and food consumption over the first two weeks of treatment, compared with Control; these reductions achieved statistical significance in animals treated at 500 and
1000 mg/kg/day. However, absolute body weights and food consumption of the females was also slightly lower than Control prior to the treatment period, and therefore this slight reduction that persisted from commencement of treatment could not be positively attributed to treatment.

Group mean body weight gains and food consumption in the males throughout the study, and for females during gestation and lactation were similar to Control.

Mating performance, fertility, duration of gestation, litter size and survival, and litter and pup weights did not indicate any obvious effect of treatment at any of the dose levels tested.

Effect levels (P0)

Key result

Dose descriptor:

NOEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Target system / organ toxicity (P0)

Key result

Critical effects observed:

no

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Details on results (F1)

VIABILITY (OFFSPRING): Number of litters with live born pups were 10, 10, 10, and 10 for the control, 100, 500 and 1000 mg/kg dose groups

CLINICAL SIGNS (OFFSPRING): No clinical signs in pups were noted.
BODY WEIGHT (OFFSPRING): Pup weights were comparable across all groups
SEXUAL MATURATION (OFFSPRING)no data
ORGAN WEIGHTS (OFFSPRING)no data
GROSS PATHOLOGY (OFFSPRING)no data
HISTOPATHOLOGY (OFFSPRING)no data

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

ca. 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

viability

clinical signs

mortality

body weight and weight gain

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:

The no observed effect level (NOEL) for adults and for reproductive parameters was 1000 mg/kg-day.

Executive summary:

OBJECTIVE: This study evaluated the potential reproduction and/or development effects following treatment with FC-770 (colorless liquid, purity 100%, lot 040031) in Sprague Dawley rats.

METHODS: This study was performed in compliance with OECD GLP. The test method was based on OECD 421. FC-770 was diluted in a vehicle composed of 0.5% Natrosol 250HX and 0.1% Tween 80 in Water for Irrigation. Rats (10/sex/group) received vehicle, 100, 500, or 1000 mg/kg-day MTDID 7145 via oral gavage. Males were dosed once daily for 4 weeks overall, commencing 2 weeks prior to mating. Females were dosed once daily from 2 weeks prior to mating and then continued until at least Day 4 of lactation. Females were euthanized with their litters between Days 5 and 6 of lactation. Parameters evaluated: Clinical observations (daily); body weights (weekly and Days 0, 7, 14, 16, 20 of gestation and Days 1 and 4 of lactation for females); food consumption (weekly and Days 0-4 of lactation for females); litter live/dead evaluation (Day 0 of lactation); necropsy; ovary and testis weights; histopathology of ovary, epididymis, and testis of control and 1000 mg/kg-day animals. Reproductive indices were calculated. Pups were examined for external abnormalities. Externally normal pups were discarded at necropsy. Externally abnormal pups were fixed in formalin for possible future analysis.

RESULTS: In treated females prior to mating, there was a slight reduction in group mean body weight gains and food consumption over the first two weeks of treatment, compared with control; these reductions achieved statistical significance in animals treated at 500 and 1000 mg/kg-day. However, absolute body weights and food consumption of the females was also slightly lower than control prior to the treatment period, and therefore this slight reduction that persisted from commencement of treatment was not likely attributed to treatment. Group mean body weight gains and food consumption in the males throughout the study, and for females during gestation and lactation were similar to control. Mating performance, fertility, duration of gestation, litter size and survival, and litter and pup weights did not indicate any obvious effect of treatment at any of the dose levels tested.

CONCLUSION: Based on the results of this study, the no observed effect level (NOEL) for adults and for reproductive parameters was considered to be 1000 mg/kg-day.