Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

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:
2012-08-29 - 2013-04-24
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline 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)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: 51 % w/w solution in water, stabilized with X % NaOH

Test animals

Species:
rat
Strain:
other: Crl: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories,Research Models and Services, Germany GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: Males/females: Both 59 days
- Weight at study initiation: Males: 272.2 g to 299.9 g, Females: 210.4 g to 233.6 g


- Fasting period before study: 16 hours
- Housing: kept individually in MAKROLON cages (type III plus)
- Diet (e.g. ad libitum): Commercial ssniff® R/M-H V1534 (ssniff Spezialdiäten GmbH, 59494 Soest, Germany; served as food ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C ± 3°C
- Humidity (%): 55% ± 15%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
VEHICLE
- Amount of vehicle (if gavage): 10 ml/kg b.w./day
Details on mating procedure:
Sexually mature male and female rats were randomly paired for mating. Mating
was monogamous: 1 male and 1 female animal were placed in one cage during the
dark period. The female was placed with the same male until pregnancy had occurred
or 2 weeks had elapsed. Each morning the females were examined for the
presence of sperm or a vaginal plug. If findings were negative, mating was repeated.
The day of conception (day 0 of gestation) was considered to be the day
on which sperm was found. In case pairing was unsuccessful, re-mating of females
with proven males of the same group was considered. This procedure was
repeated until at least 8 pregnant dams were available for each group.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For the analysis of the test item-vehicle mixtures samples of approx. 2 x 1 mL
were taken at the following time points and stored at ≤ minus 20°C until analysis
at LPT:
Start of treatment period Concentration and stability
Immediately after preparation of the test itemvehicle
mixtures as well as 8 and 24 hours after
storage of the test item preparations at room
temperature:
3 samples/dose level group (groups 2 to 4)
Number of samples: 3 x 3 = 9
Homogeneity
At start of administration, during (middle) administration
and before administration to the
last animal of each dose level group:
3 samples/dose level group (groups 2 to 4).
Number of samples: 3 x 3 = 9
End of treatment period Concentration
During treatment with the test item always before
administration to the last animal/dose level
group:
1 sample/dose level group (groups 2 to 4).
Number of samples: TD 26 (1 x 3) = 3
Number of samples: TD 39 (1 x 3) = 3
Sum of all samples: 24
Sum of all aliquots: 48
The samples were labelled with the study number, test item, test species, type of
sample, aliquot number, concentration, test day, sampling time and date.
Duration of treatment / exposure:
Males
The daily administration of the test item started
two weeks before mating and lasted until the
day before sacrifice (considering a minimum total
dosing period of at least 28 days).
Females
The daily administration of the test item started
two weeks before mating and lasted up to at
least day 3 of lactation.
Frequency of treatment:
once daily
Details on study schedule:
- Age at mating of the mated animals in the study: 10-11 week
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0 mg/kg bodyweight/day (control)
Basis:
actual ingested
Remarks:
Doses / Concentrations:
60 mg/kg bodyweight/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
200 mg/kg bodyweight/day
Basis:
actual ingested
Remarks:
Doses / Concentrations:
600 mg/kg bodyweight/day
Basis:
actual ingested
No. of animals per sex per dose:
80 animals (40 male and 40 female rats),
10 animals/sex/group.
A sufficient number to grant at least 8 pregnant
females per group for evaluation of the F0 generation
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: See supporting study: 14-DAY DOSE-RANGE-FINDING STUDY OF IBP1-Na (DANAFLOATTM 245) BY ORAL ADMINISTRATION TO RATS

Examinations

Parental animals: Observations and examinations:
AGE SIDE OBSERVATIONS: Yes
- Time schedule: at least daily, The frequency was increased when signs of toxicity were observed.
Cageside observations included skin/fur, eyes, mucous membranes, respiratory
and circulatory systems, somatomotor activity and behaviour patterns. The
onset, intensity and duration of any signs observed were recorded.
Individual animals were observed before and after dosing at each time of dosing for
any signs of behavioural changes, reaction to treatment or illness.
In addition, animals were checked regularly throughout the working day from 7:00
a.m. to 3:45 p.m. On Saturdays and Sundays animals were checked regularly from
7:00 a.m. to 11:00 a.m. with a final check performed at approximately 3:30 p.m.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: at least daily, The frequency was increased when signs of toxicity were observed.
Additionally, once before the first exposure (to allow within-subject comparisons)
and once a week thereafter, detailed clinical observations were made in all animals
outside the home cage in a standard arena and at the same time, each time preferably
by observers unaware of the treatment. Signs noted included changes in
skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and
autonomic activity (e.g. lacrimation, piloerection, pupil size, and unusual respiratory
pattern). Changes in gait, posture and response to handling as well as the
presence of clonic or tonic movements, stereotypies (e.g. excessive grooming,
repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. selfmutilation,
walking backwards) were also recorded.
Dated and signed records of appearance, change, and disappearance of clinical
signs were maintained on clinical history sheets for individual animals.

BODY WEIGHT: Yes
- Time schedule for examinations: Males and females were weighed on the first day of dosing, weekly thereafter and
at termination of the study. During gestation, females were weighed on days 0, 7,
14 and 20 and within 24 hours of parturition (day 1 post-partum) and day 4 postpartum.
Body weights were recorded individually for each adult animal.

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
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
No test item-related premature death was noted
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
A slight reduction in body weight was noted until the end of the study on test day 38 for the male rats of the high dose group (600 mg IBP1-Na/kg b.w./day). Body weight at autopsy was reduced accordingly
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
A slight reduction in body weight was noted until the end of the study on test day 38 for the male rats of the high dose group (600 mg IBP1-Na/kg b.w./day). Body weight at autopsy was reduced accordingly
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
The histopathological examination revealed test item related changes in the forestomach from male and female rats of the intermediate and the high dose group (200 and 600 mg IBP1-Na/kg b.w./day) in form of hyperplasia and hyperkeratosis of the squamous ce

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)

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: General health parameters
Key result
Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: Reproductive parameters

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In the high dose group the mean litter weight of the male and female pups was decreased by 5.6% on lactation day 1 and by 9.6% on lactation day 4 (without statistical significance). No noticeable differences were noted for the total litter weight
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

Effect levels (F1)

Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: General health parameters

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
The aim of the experiment was to obtain information on possible effects of the test item IBP1-Na on reproduction and/or development according to OECD guideline 422. The test item was administered orally to rats at dose levels of 60, 200 or 600 mg IBP1-Na/kg b.w./day.
The application started two weeks before mating on test day one and ended on the day or one day before sacrifice. Day of sacrifice was on test day 38 for the male rats and between lactation day 4 and 5 for the female rats.

Effects on the parental generation
No test item-related premature death was noted.
At the intermediate and the high dose level (200 and 600 mg IBP1-Na/kg b.w./day) a slight to extreme salivation was noted in nearly all animals, with a higher incidence at the high dose level. Piloerection and an increased water consumption were noted for a few animals of the intermediate and the high dose group.
The neurological screening revealed a reduction for the hindlimb grip strength of the male and female animals of the high dose group (600 mg IBP1-Na/kg b.w./day).
A slight reduction in body weight was noted until the end of the study on test day 38 for the male rats of the high dose group (600 mg IBP1-Na/kg b.w./day). Body weight at autopsy was reduced accordingly.
Examination of the haematological and biochemical parameters revealed statistically significant changes in the male and female animals of the high dose group (600 mg IBP1-Na/kg b.w./day), which were considered as to be test item-related.
Macroscopic inspection at autopsy revealed test item related changes in the cardiac part of the stomach from 3 male animals of the intermediate dose group (200 mg IBP1-Na/kg b.w./day) and in the cardiac part of the stomach from 1 female animal of the high dose group (600 mg IBP1-Na/kg b.w./day).
Examination of the organ weights revealed a slightly statistically significant increase in the relative liver weight of the male rats of the high dose group (600 mg IBP1-Na/kg b.w./day).
The histopathological examination revealed test item related changes in the forestomach from male and female rats of the intermediate and the high dose group (200 and 600 mg IBP1-Na/kg b.w./day) in form of hyperplasia and hyperkeratosis of the squamous cell epithelium, infiltration with neutrophilic granulocytes, granulation tissue and ulcerations.

Additionally, a test item-related centrilobullar hepatocellular hypertrophy was noted In the liver from male and female animals of the high dose group (600 mg IBP1-Na/kg b.w./day).

Effects on reproduction parameters and organs
No test item-related influence was noted on the reproduction parameters and organs in any treatment group.
The qualitative sperm staging revealed no test item-related spermatogenic changes.

Effects on the development of the F1 offspring (pups)
A non-statistically significant reduction in mean litter weight of approximately 6% to 9% was noted on lactation day 1 and 4 for the male and female pups of the high dose group (600 mg IBP1-Na/kg b.w./day), which is regarded to be test item-related.
No test item related influence was noted on the survival rate of the pups.

The following no-observed-effect levels were established for systemic effects:
Effects on the F0-generation
NOAEL (no-observed-adverse-effect level): 200 mg IBP1-Na/kg b.w./day, p.o.

Effects on reproduction
NOAEL (no-observed-adverse-effect level): 200 mg IBP1-Na/kg b.w./day, p.o.

Remark: The test solution contains NaOH due to the production method.