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Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
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:
11 August 2011 - 08 March 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was performed according to OECD test guidelines, and in compliance with GLP, so the data is considered reliable without restriction.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: Approximately 72 days
- Weight at study initiation: 348 - 399g (males), 230 - 287g (females)
- Housing: Polycarbonate cages with either stainless steel grid floors during mating, and solid poly carbonate floors during other phases of testing.
- Diet (e.g. ad libitum): ad libitum / free access, except overnight before routine blood sampling
- Water (e.g. ad libitum): ad libitum / free access
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 23°C
- Humidity (%): 40 to 70%
- Photoperiod (hrs dark / hrs light): 12 hours light: 12 hours dark

IN-LIFE DATES: From: 26 October 2011 (Treament commenced) To: 12 December 2011 (Last date of necropsy for main phase females)
Route of administration:
oral: gavage
Vehicle:
other: 1% (w/v) aqueous methylcellulose
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Concentration in vehicle: 10 - 100 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw/day
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: Up to two weeks, or until evidence of mating was found
- Proof of pregnancy: vaginal plug or sperm in vaginal smear referred to as day 0 of gestation
- After successful mating each pregnant female was caged (how): Individually
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Formulation samples were analysed by Atomic Absorption Spectrometry, to determine Iron in the samples.

Prior to the start of treatment, the analytical method was validated with respect to specificity, limit of detection, linearity of detector response over the calibration range, precision of measurement at the lowest and highest calibration standards, and the accuracy and precision of the method, by the determination of six procedural recoveries at 1 mg/mL and 100 mg/mL. A stability tiral was also performed; formulations were found to be stable and homogenous for up to 2 hours at ambient temperature with paddle stirring, and following 15 days' refrigerated storage.

Samples from all formulations prepared in the first and last weeks of the study were analysed; the test material concentrations were found to be within acceptable limits, confirming accurate preparation.
Duration of treatment / exposure:
Main phase males and toxicity phase females were dosed for five consecutive weeks.
Main phase females were treated daily for two weeks before pairing, throughout mating, gestation and until day 6 of lactation.
Frequency of treatment:
Daily
Details on study schedule:
- Age at mating of the mated animals in the study: Approximately 12 weeks
Remarks:
Doses / Concentrations:
100 mg/kg/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
300 mg/kg/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/kg/day
Basis:
actual ingested
No. of animals per sex per dose:
Main Phase - 10 males and 10 females per dose.
Toxicity phase - 5 females per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Dose levels were decided in a 7-day preliminary study in rats, conducted at the same laboratory - refer to "7 Day rangefinding_Huntingdon Life Sciences, 2011 (FGE0026)".

Parental animals: Observations and examinations:
Refer to Study Summary in section 7.5.1.
Oestrous cyclicity (parental animals):
For 15 days before pairing, daily vaginal smears (dry) were taken from all Main phase females, using cotton swabs moistened with saline. The smears were subsequently examined to establish the duration and regularity of the oestrous cycle. After pairing with the Main phase male, smearing was continued using pipette lavage, until evidence of mating was observed.
Sperm parameters (parental animals):
Parameters examined in male parental generations: testis weight, epididymis weight.
Seminal vesicles were subject to histopathological examination.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: No

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring:
Clinical signs, litter size, Sex ratio, and bodyweight

GROSS EXAMINATION OF DEAD PUPS:
For offspring surviving to scheduled termination, a careful external examination for gross abnormalities was performed. Offspring that appeared normal externally were discarded without internal examination. Offspring which were externally abnormal were subjected to a full internal microscopic examination. Missing offspring and those grossly autolysed or cannibalised could not be examined; all other offspring which died before day 7 of age were examined as detailed. The necropsy also included an assessment for the presence of milk in the stomach, where possible.
Postmortem examinations (parental animals):
Refer to Gross Pathology and Histopathology in section 7.5.1.
Postmortem examinations (offspring):
Microscopic examination, as described in "Litter Observations", above.
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:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
not examined
Reproductive function: oestrous cycle:
no effects observed
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
no effects observed
Refer to detailed analysis of toxicological parameters in section 7.5.1.

REPRODUCTIVE FUNCTION: ESTROUS CYCLE (PARENTAL ANIMALS)
Assessment of oestrous cycles during the two week pre-pairing period showed that nearly all of the main phase females had regular 4 or 4/5 day oestrous cycles and it was considered that this parameter was not affected by treatment with Fe3P.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
The seminiferous tubules of the testes were evaluated with respect to their stage in the spermatogenic cycle and the integrity of the various cell types present within the different stages. No cell or stage abnormalities were noted.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
The pre-coital interval was unaffected by treatment, with animals mating at the earliest opportunity, when the female came into oestrus.
Mating performance and fertility as assessed by percentage mating, conception rate and fertility index, were unaffected by treatment.
The gestation length was within the normal range of 22 to 23 days for all animals and the gestation index was unaffected by treatment.
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Sexual maturation:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
not examined
VIABILITY (OFFSPRING)
The mean number of implantations and the live litter size on Days 1, 4 and 7 were similar to control values and unaffected by treatment.
Offspring survival up to Day 7 of age was unaffected by parental treatment.

CLINICAL SIGNS (OFFSPRING)
Refer to table in "Any other information on results".

BODY WEIGHT (OFFSPRING)
Mean bodyweights of male and female offspring of all groups of parents treated with Fe3P were marginally lower than in Controls on Day 1 of age but differences did not attain statistical significance and there was considered to be no effect of treatment on mean bodyweight gains of the offspring between Days 1 and 4 of age. However, between Day 4 and Day 7 of age, mean bodyweight gains of male and female offspring of all groups of parents treated with Fe3P were lower than in Controls and the differences attained statistical significance in the 1000 or 300 mg/kg/day groups (77-80% of Controls) and they showed a dose response. Overall bodyweight gain, between Days 1-7, was statistically lower in all male offspring of all groups of parents treated with Fe3P.

GROSS PATHOLOGY (OFFSPRING)
Macroscopic examination of offspring dying before scheduled termination or killed at scheduled termination on Day 7 of age did not reveal any findings that were attributed to parental treatment.

OTHER FINDINGS (OFFSPRING)
Sex ratio, as expressed in terms of % males, was not affected by parental treatment.
Reproductive effects observed:
not specified

Summary of offspring clinical observations

Observation

Group (Dose mg/kg/day)

1 (0)

2 (100)

3 (300)

4 (1000)

Number of offspring (litters) affected

Head: bruising and swelling

1(1)

 

 

 

Head: bruising

 

1(1)

1(1)

1(1)

Umbilical cord attached

1(1)

 

 

 

Missing

2(2)

 

1(1)

2(2)

Tail absent

1(1)

 

 

 

Dorsal surface: Scab and reddening

 

1(1)

 

 

Found dead

 

1(1)

 

 

Forepaw: Scab

 

1(1)

 

 

Colour: Pale

 

 

1(1)

 

Conclusions:
The No Observed Adverse Effect Level (NOAEL) for Fe3P for reproductive and developmental effects was considered to be 1000 mg/kg/day.
Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

A combined repeat dose toxicity study and reproductive / developmental toxicity screening study in rats was conducted (Huntingdon Life Sciences, 2012) to assess the effect of Fe3P on rats following repeated oral administration. The study was conducted according to OECD test guideline 422, and in compliance with GLP.

Male and female rats were administered Fe3P by oral gavage for five weeks (males, and toxicity phase females), or for two weeks before mating, during mating and gestation, and until day six of lactation (main phase females). The dose groups were 100, 300, and 1000 mg/kg/day, and a concurrent control group was administered with untreated vehicle.

Reproductive performance was not affected by treatment with test material; the assessed factors included mating performance, fertility and offspring survival. No adverse effects were seen on offspring development up to Day 7 of age.

On the basis of this study, the No Observed Adverse Effect Level for reproductive and developmental toxicity is considered to be 1000 mg/kg/day.


Short description of key information:
NOAEL, rat, oral exposure = 1000 mg/kg/day

Effects on developmental toxicity

Description of key information
NOAEL, rat, oral exposure = 1000 mg/kg/day
Link to relevant study records
Reference
Endpoint:
developmental toxicity
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Abnormalities:
not specified
Developmental effects observed:
not specified
Effect on developmental toxicity: via oral route
Endpoint conclusion:
no study available
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

Refer to fertility discussion for details.

Justification for classification or non-classification

In the above reproductive / developmental toxicity screening study, no effects were observed on the reproduction of rats following repeated exposure to Fe3P, and no adverse effects were seen on the development of offspring. There is therefore no basis on which to justify calssification of Fe3P for reproductive or developmental toxicity.