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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on test type (migrated information)
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
1993-06-01 to 1993-11-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study is classified as reliable with restrictions because it was compliant with GLPs and appeared to follow OECD 415 guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 415 [One-Generation Reproduction Toxicity Study (before 9 October 2017)]
Principles of method if other than guideline:
This study was the preliminary part of a 91-day subchronic study with in utero exposure. Males and females were mated for 4 weeks prior to mating and through mating until sacrifice, which for females included through gestation and lactation day 20.
GLP compliance:
yes
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Ethylflo 166 poly alpha olefin
- Substance type: Poly alpha olefins (1-decene homopolymer hydrogenated)
- Physical state: Liquid
- Analytical purity: Not reported
- Composition of test material, percentage of components: C30=31.27%; C40=45.02%; C50=17.44%; C60=6.27%
- Lot/batch No.: 200-135 and 200-185
- Expiration date of the lot/batch: Not reported
- Stability under test conditions: Not reported
- Storage condition of test material: Room temperature
- Other: Chemical identity, purity, strength, and stability were stated to be the responsibility of the sponsor

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Raleigh, North Carolina
- Age at study initiation: (P) 10 weeks; The F1 generation was used for a separate subchronic study and not reproduction
- Weight at study initiation: (P) Males: 319 to 422 grams; Females: 202 to 274 grams; F1 animals were not used as a part of the reproduction study
- Housing: Individually except during cohabitation
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 17 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24
- Humidity (%): 40 to 70%
- Air changes (per hr): 10 to 12 per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light


IN-LIFE DATES: From: 1995-06-01 To: 1993-08-11

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Specific amount of test compound was weighed into a beaker with the appropriate amount of vehicle. The mixture was stirred by hand then sufficient vehicle was added to achieve the desired concentration. Solutions were then stirred for 30 minutes using a stir bar.


VEHICLE
- Justification for use and choice of vehicle (if other than water): Not reported
- Concentration in vehicle: 0, 20, 100, or 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg of vehicle or solution was administered
- Lot/batch no. (if required): 920897 and 933384
- Purity: Not reported
Details on mating procedure:
- M/F ratio per cage: 1 to 1 ratio
- Length of cohabitation: 15 days
- Proof of pregnancy: Vaginal plug or sperm in vaginal smear referred to as day 0 of pregnancy
- After 10 days of unsuccessful pairing replacement of first male by another male from the same group.
- Further matings after two unsuccessful attempts: No
- After successful mating each pregnant female was caged in nesting boxes
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Prior to study initiation, homogeneity and stability tests were performed. All test mixtures were analysed for verification of the test article concentration. Samples were found to be homogeneous and stable. All concentrations used were found to be within 20% of the nominal concentration.
Duration of treatment / exposure:
Males and females were dosed for 4 weeks prior to mating and through mating until sacrifice, which for females included through gestation and lactation day 20.
Frequency of treatment:
Daily
Details on study schedule:
- F1 animals not mated.
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 500, or 1000 mg/kg/day
Basis:
other: nominal in polyethylene glycol 400
No. of animals per sex per dose:
Thirty animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Not reported
- Rationale for animal assignment (if not random): Random
Positive control:
No positive control was used.

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least once a day
- Cage side observations included mortality, morbidity, and overt signs of toxicity.


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly


BODY WEIGHT: Yes
- Time schedule for examinations: Males: weekly; Females: Gestational days 0, 7, 14, and 20 and lactation days 1, 7, 14, and 21


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


WATER CONSUMPTION: No


Oestrous cyclicity (parental animals):
Estrous cyclicity was not examined.
Sperm parameters (parental animals):
Sperm parameters were not examined.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: Yes
- If yes, maximum of eight pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.


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, and clinical signs


GROSS EXAMINATION OF DEAD PUPS:
Yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals at the end of breeding.
- Maternal animals: All surviving animals on lactation day 21.


GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.


HISTOPATHOLOGY / ORGAN WEIGHTS: Only abnormal tissues were preserved for possible microscopic examination. No organs were specified as weighed.
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected for subchronic study were sacrificed at 21 days of age.
- These animals were subjected to postmortem examinations macroscopic examination as follows:

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.


HISTOPATHOLOGY / ORGAN WEIGHTS: No histopathology or organ weights were conducted.
Statistics:
Continuous data were analysed using a one way analysis of variance followed by a Dunnett's test (sometimes a modified Dunnett's test). Count data were analysed using a Chi-square or a Mann-Whitney U test.
Reproductive indices:
Gestation and lactation body weight, copulation index, fertility index, precoital interval, gestation length, gravid and non-gravid females that did not deliver, and presence of corpora lutea
Offspring viability indices:
Alive pups on day 1, 4, 8, 14, and 21; dead and live pups on lactation day 0; sex ratio; mean litter size; and body weight

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
not examined
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS): Soft stools and faecal staining occurred in all groups including the control and were considered related to the vehicle used. There were no clinical signs related to treatment.


BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS): There were no treatment-related effects.


REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS): There were no treatment-related effects.


GROSS PATHOLOGY (PARENTAL ANIMALS): There were no treatment-related effects.


Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: overall effects

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 examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Histopathological findings:
not examined

Details on results (F1)

VIABILITY (OFFSPRING): There were no treatment-related effects on the viability parameters measured.


CLINICAL SIGNS (OFFSPRING): There were no clinical signs related to treatment.


BODY WEIGHT (OFFSPRING): There were no treatment-related effects on pup body weight.


GROSS PATHOLOGY (OFFSPRING): There were no gross pathology findings related to treatment.


Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
Ethylflo 166 did not appear to have any effects on reproduction.
Executive summary:

In a one-generation reproduction study, ethylflo 166 was administered to 30 Sprague-Dawley Crl:CD®BR VAF/Plus® rats/sex/dose by gavage at dose levels of 0, 100, 500, or 1000 mg/kg bw/day. Both males and females were treated for 4 weeks prior to mating and through mating. At the end of mating, males were sacrificed. Females were treated through gestation and until lactation day 21. On lactation day 4, litters were culled to 8 pups (4 per sex).

 

There were no treatment-related effects on clinical signs, mortality, body weight, or gross pathology in the parental generation or in the pups through lactation day 21. There were no treatment related effects on reproduction or pup viability. Some pups were used further in a subchronic study with the remainder sacrificed on lactation day 21. There is no parental or offspring systemic or reproduction LOAEL, based n the lack of effects. The parental systemic and reproduction NOAEL is 1000mg/kg bw/day in males and females. The offspring NOAEL is 1000 mg/kg bw/day even after the additional 91 day subchronic exposure.

 

This study received a Klimisch score of 1 and is classified as reliable with restrictions because it was compliant with GLPs and appeared to follow OECD 415 guidelines. It should be noted that this study provides information on the reproduction in rats, but was part of a subchronic study with exposure in utero and therefore did not examine all aspects of reproduction. Consequently, it does not meet REACh's requirement for a two-generation study. This study will not influence the DNEL.