3-ethoxy-1,1,1,2,3,4,4,5,5,6,6,6-dodecafluoro-2-(trifluoromethyl)-hexane

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:

20 November 2009 to 15 February 2010

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Deviations:

yes

Remarks:

a very minor deviation in clinical observation reporting was occurred which did not effect study interpretation

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

435-790-1

EC Name:

-

Cas Number:

297730-93-9

Molecular formula:

C9H5F15O

IUPAC Name:

3-ethoxy-1,1,1,2,3,4,4,5,5,6,6,6-dodecafluoro-2-(trifluoromethyl)hexane

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River UK Limited, Margate, Kent, UK
- Age at study initiation: 7 weeks
- Weight at study initiation: 169-187 g for males and 143-160 g on receipt from supplier. Testing commenced two weeks later
- Fasting period before study: No
- Housing:
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: 13 days prior to treatment
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 2 deg C. (17-23 measured)
- Humidity (%): 55+/- 15% (35-62% measured)
- Air changes (per hr): 15+
- Photoperiod (hrs dark / hrs light): 12 hours/12 hours
IN-LIFE DATES: From: To: November 2009 to January 18, 2010

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 20% D-alpha-Tocopherol polyethylene glycol succinate (TPGS, Vitamin E) in water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:


VEHICLE
- Justification for use and choice of vehicle (if other than water):

Selected vehicle was used to assure proper reproducible dosing of vehicle.

- Concentration in vehicle:
- Amount of vehicle (if gavage): 3mL per kg of body weight
- Lot/batch no. (if required):
- Purity:



GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:
- Method of holding animals in test chamber:
- Source and rate of air:
- Method of conditioning air:
- System of generating particulates/aerosols:
- Temperature, humidity, pressure in air chamber:
- Air flow rate:
- Air change rate:
- Method of particle size determination:
- Treatment of exhaust air:
TEST ATMOSPHERE
- Brief description of analytical method used:
- Samples taken from breathing zone: yes/no


VEHICLE (if applicable)
- Justification for use and choice of vehicle:
- Composition of vehicle:
- Type and concentration of dispersant aid (if powder):
- Concentration of test material in vehicle:
- Lot/batch no. of vehicle (if required):
- Purity of vehicle:



TEST SITE
- Area of exposure:
- % coverage:
- Type of wrap if used:
- Time intervals for shavings or clipplings:
REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:



TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution): 333mg/mL, 66.7 mg/mL, 13.3 mg/mL
- Constant volume or concentration used: Constant concentrations
- For solids, paste formed: yes/no


VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity:




USE OF RESTRAINERS FOR PREVENTING INGESTION: yes/no

Details on mating procedure:

- M/F ratio per cage: one-to-one

- Length of cohabitation: Until mated ir until 14 days

- Proof of pregnancy:
From results of evaluation of vaginal lavage

- Further matings after two unsuccessful attempts: [no / yes (explain)] n.a.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The formulations were prepared at convenient intervals and were used within the 8 days stability period (established in Charles River Study No. 425881) at 2-8°C in the dark. The test item was formulated by adding the appropriate volume of vehicle (20% TPGS) to required amounts of test item and mixed manually and magnetically stirred until a visibly homogeneous suspension was obtained.

Duration of treatment / exposure:

4-weeks for males starting 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; the females were sacrificed with their litters between Days 5 and Day 7 of lactation.

Frequency of treatment:

daily

Details on study schedule:

- Age at mating of the mated animals in the study: [...10 weeks

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 40, 200, 1000 mg/kg/day
Basis:
nominal conc.

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Based upon toxicity data from earlier studies.

Positive control:

None

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes / No / No data
Yes

- Time schedule: Daily
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes

- Time schedule: Daily

BODY WEIGHT:

Yes

- Time schedule for examinations: Daily

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data

Yes

Sperm parameters (parental animals):

Parameters examined in [all/P/F1/F2] male parental generations:
[testis weight, epididymis weight, daily sperm production, sperm count in testes, sperm count in epididymides, enumeration of cauda epididymal sperm reserve, sperm motility, sperm morphology, other:]

Sperm examined in the lavaged samples

Litter observations:

STANDARDISATION OF LITTERS

- Performed on day 4 postpartum: Yes

- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); excess pups were killed and discarded.


PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] 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:
[no / yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead.]

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving animals
- Maternal animals: All surviving animals
GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]
HISTOPATHOLOGY / ORGAN WEIGHTS
Histological examination was conducted on Control and High dose adult animals only. A section of each ovary and each epididymis, and a transverse section from each testis were stained with Haematoxylin and Eosin (H&E) and a further section from each testis was stained with PAS-Haematoxylin.

The following organs were examined:
Ovaries
Uterus, cervix and vagina
Testes (weighed individually)
Epididymides (weighed individually)
Seminal vesicles and coagulating gland,
Prostate gland
Pituitary gland

Postmortem examinations (offspring):

SACRIFICE
Offspring killed or found dead were sexed, 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:

Fertility index (male and female), gestation index

Offspring viability indices:

Birth index, live birth index, viability index

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Description (incidence and severity):

There were no clinical observations that were considered to be related to treatment with the test article.

Mortality:

mortality observed, non-treatment-related

Description (incidence):

At 0 mg/kg/day, one female (Animal 45) was killed prematurely on Day 23 of study following a dosing error. This animal swallowed part of the gavage during the dosing procedure; at necropsy part of the gavage tube was found in the oesophagus.

Body weight and weight changes:

no effects observed

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

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Other effects:

no effects observed

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Reproductive function: sperm measures:

not examined

Reproductive performance:

no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
There were no clinical observations or necropsy findings that were considered to be related to treatment with the test article.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
Group mean body weight gain in treated males was similar to Control. No significantchanges were identified in the treated group mean values compared to Control. For females prior to mating, at 1000 mg/kg/day, group mean body weight gain over Week 0-2
was higher compared to Control, attaining statistical significance. However, this significance was achieved due to a low body weight gain for Control animals 42 and 43 over the first week
of treatment. Excluding these Control animals, group mean body weight gain was essentially similar to Control. Group mean body weight gains during gestation and lactation were similar
to Control.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)

There were no obvious effects of treatment on mating performance, fertility or duration of gestation.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
There were no obvious effects of treatment on mating performance, fertility or duration of gestation.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)

There were no obvious effects of treatment on mating performance, fertility or duration of
gestation.

ORGAN WEIGHTS (PARENTAL ANIMALS)
The weights of the male reproductive organs were essentially similar in all groups.


GROSS PATHOLOGY (PARENTAL ANIMALS)

HISTOPATHOLOGY (PARENTAL ANIMALS)
There were no histology findings following examination of the ovaries from the females or the epididymides and testes of the males that were considered to be related to treatment with the test article.

Effect levels (P0)

Dose descriptor:

NOEL

Effect level:

> 1 000 mg/kg bw/day

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Remarks on result:

other: Generation: Parental and offspring (migrated information)

Results: P1 (second parental generation)

Effect levels (P1)

Key result

Dose descriptor:

NOEL

Effect level:

ca. 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

clinical signs

mortality

body weight and weight gain

food consumption and compound intake

organ weights and organ / body weight ratios

histopathology: non-neoplastic

reproductive performance

other: Mating performance, fertility, duration of gestation and litter size were not impacted by treatment with the test article at dose levels up to 1000 mg/kg/day. The NOEL is 1000 mg/kg/day.

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 specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

Details on results (F1)

VIABILITY (OFFSPRING)
There were no obvious effects of treatment on litter size, implant count or survival.
CLINICAL SIGNS (OFFSPRING) No adverse effects
BODY WEIGHT (OFFSPRING)
Mean litter and pups weights were essentially similar in all groups.
SEXUAL MATURATION (OFFSPRING)
ORGAN WEIGHTS (OFFSPRING)
GROSS PATHOLOGY (OFFSPRING)
HISTOPATHOLOGY (OFFSPRING)
OTHER FINDINGS (OFFSPRING): None

Effect levels (F1)

Key result

Dose descriptor:

NOEL

Generation:

F1

Effect level:

ca. 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

viability

body weight and weight gain

gross pathology

other: VIABILITY: There were no obvious effects of treatment on litter size, implant count or survival. CLINICAL SIGNS: No adverse effects BODY WEIGHT: Mean litter and pups weights were essentially similar in all groups. The NOEL is 1000 mg/kg/day.

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Applicant's summary and conclusion

Conclusions:

The no observed effect level (NOEL) for both parental and reproductive effects was 1000 mg/kg/day.

Executive summary:

At levels up to 1000 mg/kg/day of MTDID 7910, there were no obvious effects of treatment on clinical observations, necropsy findings, body weight gain or food consumption. Mating performance, fertility, duration of gestation, litter size and survival, and litter and pup weights did not indicate any obvious adverse effect of treatment at any of the dose levels tested. Under the conditions of this study, the no observed effect level (NOEL) for both parental and reproductive effects was 1000 mg/kg/day.