Registration Dossier

Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on generations indicated in Effect levels (migrated information)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2013, January-May
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP study, well conducted on the similar substance Reaction mass of dipotassium phosphonate and Phosphonic acid, potassium salt (1:1). Rationale for Read Across is attached at point 13
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Batch number : 20111216-014
Date of expiry : 13 December 2013
Appearance : White clear fluid solution
Storage conditions : Room temperature

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Animal supply and acclimatisation
A total of 90 Sprague Dawley rats (45 males and 45 virgin females), 6 to 7 weeks old and weighing 176 to 200 g for males and 151 to 175 g for females, have been ordered from Charles River Italia S.p.A., Calco (Lecco), Italy.
After arrival the weight range for each sex have been determined and the animals have been temporarily identified within the cage by means of a coloured mark on the tail. A health check has then been performed by a veterinarian.
An acclimatisation period of approximately 2 weeks has been allowed before the start of treatment, during which time the health status of the animals has been assessed by thorough observations. Rats considered unsatisfactory have been killed and were appropriate subjected to pathological examination.

Animal husbandry
The animals have been housed in a limited access rodent facility. Animal room controls have been set to maintain temperature and relative humidity at 22°C +/- 2°C and 55%+/- 15% respectively; actual conditions have been monitored, recorded and the records retained. There will be approximately 15 to 20 air changes per hour and the rooms will be lit by artificial light for 12 hours each day.
From arrival to pairing, animals have been housed up to 5 of one sex to a cage, in polisulphone solid bottomed cages measuring 59.5x38x20 cm (Techniplast Gazzada S.a.r.l., Buguggiate, Varese). Nesting material has been provided inside suitable bedding bags and changed at least twice a week.
During mating, animals have been housed one male to one female in clear polycarbonate cages measuring approximately 43x27x18cm with a stainless steel mesh lid and floor (Techniplast – Gazzada S.a.r.l.). Each cage tray holded absorbent material which has been inspected and changed daily.
After mating, the males have been recaged as they were before mating, the females have been transferred to individual solid bottomed cages (Techniplast Gazzada S.a.r.l.) for the gestation period, birth and lactation. Suitable nesting material has been provided and has been changed as necessary.
Drinking water has been supplied ad libitum to each cage via water bottles, except in the case of urinalysis investigations.
A commercially available laboratory rodent diet (4 RF 21, Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI), Italy) will be offered ad libitum throughout the study.
There is no information available to indicate that any non-nutrient substance likely to influence the effect of the test item is present in the drinking water or the diet. Records of analyses of water and diet are kept on file at RTC.
Dated and signed records of activities relating to the day to day running and maintenance of the study in the animal house will be recorded.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
Males
Animals have been dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter through the day before necropsy.
Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.

Females
Animals will be dosed once a day, 7 days a week, for a minimum of 2 consecutive weeks prior to pairing and thereafter during pairing , post coitum and post partum periods until Day 3 post partum or the day before sacrifice. Dose volumes will be adjusted once per week for each animal according to the last recorded body weight.
During the gestation period, dose volumes will be calculated according to individual body weight on Days 0, 7, 14 and 20 post coitum and on Day 1 post partum. Thereafter individual dose volumes will remain constant.
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
10 mg/Kg bw
Basis:
actual ingested
Remarks:
Doses / Concentrations:
100 mg/Kg bw
Basis:
actual ingested
Remarks:
Doses / Concentrations:
1000 mg/Kg bw
Basis:
actual ingested
No. of animals per sex per dose:
10 animal per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
The test item have been administered orally by gavage at a dose volume of 10 mL/kg body weight. Control animals received the vehicle alone at the same dose volume.
The dose has been administered to each animal on the basis of the most recently recorded body weight and the volume administered has been recorded for each animal

Examinations

Parental animals: Observations and examinations:
Mortality
Early in each working day in the morning and in the afternoon. At weekends and Public Holidays a similar procedure will be followed except that the final check has been carried out at approximately mid-day.

Clinical signs
Once before commencement of treatment and at least once daily during the study,

Clinical observations (Functional Observation Battery Tests)
Once before commencement of treatment and at least once a week thereafter

Grip strength and sensory reactivity to stimuli
Once during the study, towards the end of treatment, on 5 males and 5 females randomly selected from each group

Motor activity assessment (MA)
Once during the study, towards the end of treatment, on 5 males and 5 females randomly selected from each group

Food consumption
Recorded weekly (whenever possible) during the pre-mating period starting from allocation. Individual food consumption for the females have been measured on gestation Days 7, 14 and 20 starting from Day 0 post coitum and on Day 4 post partum starting from Day 1 post partum.

Body weight
Males weekly from allocation to termination.
Females weekly from allocation to positive identification of mating and on gestation Days 0, 7, 14 and 20.
Dams Days 1 and 4 post partum.
Oestrous cyclicity (parental animals):
Vaginal smears
Vaginal smears has been taken daily in the morning starting two weeks before pairing until a positive identification of copulation is made.
Litter observations:
Pups identification, weight and observation
As soon as possible, after parturition is considered complete (Day 0 or 1 post partum), .
Once daily for all litters

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:
no effects observed
Gross pathological findings:
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:
no effects observed
Reproductive performance:
no effects observed

Effect levels (P0)

Dose descriptor:
NOEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female

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:
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed

Effect levels (F1)

Dose descriptor:
NOEL
Generation:
F1
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
NOEL oral rat > 1000 mg/Kg bw