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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose for cross-reference:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
adopted: 27.07.1995
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
Landesamt für Umwelt, Wasserwirtschaft und Gebäudeaufsicht, Mainz
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
-
EC Number:
485-430-0
EC Name:
-
Cas Number:
923954-49-8
Molecular formula:
SnxZnyTi2Oz with x = 0,5 – 1,7 y = 0,5 – 1,7 z = 5,5 – 7,5
IUPAC Name:
Tin Titanium Zinc kegginite B
Details on test material:
- Name of test substance: Tin titanium zinc oxide
- CAS No.: 923954-49-8
- Test substance No.: 07/0285-1
- Batch identification GV 33900-66
- Purity: 99.61g / 100g (Analytical report: 07L00144)
- Homogeneity: given
- Stability: Unlimited. The stability under storage conditions over the prospective administration period was guaranteed by the manufacturer, and the manufacturer holds this responsibility.
- Physical state/appearance: solid/orange
- Storage conditions: room temperature

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Age at study initiation: 10-11 weeks (males/females)
- Weight at study initiation: (P) Males: 316.6-342.8 g; Females: 184.4-216.0 g
- Housing: During the study period, the rats were housed individually in Makrolon type M III cages (Becker & Co., Castrop-Rauxel, Germany), with the following exceptions: • During overnight mating, male and female mating partners were housed together in Makrolon type M III cages. • Pregnant animals and their litters were housed together until PND 4 (end of lactation).
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%
- Air changes (per hr): 15 times per hour
- Photoperiod (hrs dark / hrs light): 12 hours / 12 hours

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
1% in water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Concentration in vehicle: 1% carboxymethylcellulose in drinking water
Details on mating procedure:
- M/F ratio per cage: 1:1 ratio
- Length of cohabitation: Each of the male and female animals was mated overnight for a maximum of 2 weeks. The animals were paired by placing the female in the cage of the male mating partner from about 16.00 h until 07.00-09.00 h of the following morning.
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The analytical investigations of the test substance preparations was carried out at Competence Center Analytics, Department of BASF SE, Ludwigshafen, Germany, as a separate GLP study.
At the beginning of the administration period samples were taken from the lowest and highest concentration for homogeneity analysis (were also used for concentration control). From the mid concentration a concentration control analysis was carried out. At the end of the administration period concentration control analyses of the test substance preparations were performed in samples of all concentrations.
Duration of treatment / exposure:
51 days F0 females and 44 days F0 males : The treatment lasted up to one day prior to sacrifice.
Frequency of treatment:
daily, at the same time in the morning
Doses / concentrationsopen allclose all
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once daily

BODY WEIGHT: Yes
- Time schedule for examinations: once a week at the same time of the day (in the morning). During gestation and lactation period, F0 females were weighed on gestation days (GD) 0, 7, 14 and 20, on the parturition day (postnatal day [PND] 0) and on PND 4.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption was determined once a week (in a period of 7 days) for male and female parental animals, with the following exceptions:
• Food consumption was not determined after the 2nd premating week (male parental animals) and during the mating period (male and female F0 animals). • Food consumption of the F0 females with evidence of sperm was determined on GD 0, 7, 14 and 20. • Food consumption of F0 females, which gave birth to a litter, was determined on PND 1 and 4.
Food consumption was not determined in females without positive evidence of sperm and females without litter.
Litter observations:
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, physical or behavioural abnormalities

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.

CLINICAL OBSERVATIONS
The live pups were examined daily for clinical symptoms (including gross-morphological findings) during the clinical inspection of the dams.

BODY WEIGHT DATA
The pups were weighed one day after birth (PND 1) and on day 4 after birth.
Postmortem examinations (parental animals):
GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations and assessed by gross pathology, special attention being given to the reproductive organs.

HISTOPATHOLOGY / ORGAN WEIGHTS
Weight assessment was carried out on all animals sacrificed at scheduled dates. The following weights were determined:
1. Anesthetized animals, 2. Epididymides, 3. Ovaries, 4. Testes.
Histopathologycal examination by light microscopy.
Postmortem examinations (offspring):
SACRIFICE
- On PND 4 all pups were sacrificed under isoflurane anesthesia with CO2 and examined macroscopically. Stillborn pups and all pups which died ahead of schedule were examined externally, eviscerated and their organs were assessed macroscopically.

GROSS NECROPSY
- All surviving pups (sacrificed on PND 4 under isoflurane anesthesia with CO2), all stillborn pups and those pups, which died ahead of schedule, were examined externally, eviscerated and their organs were assessed macroscopically.
Statistics:
Statistics of clinical examinations:
Food consumption (parental animals), body weight and body weight change (parental animals and pups; for the pup weights, the litter means were used), number of mating days, duration of gestation, number of pups delivered per litter, implantation sites, post implantation loss: Simultaneous comparison of all dose groups with the control group using the DUNNETT-test (two-sided) for the hypothesis of equal means.
Male and female mating index, male and female fertility index, gestation index, females with liveborn pups, females with stillborn pups, females with all stillborn pups, live birth index, pups stillborn, pups died, pups cannibalized, pups sacrificed moribund, viability index, number of litters with affected pups at necropsy: Pairwise comparison of each dose group with the control group using FISHER'S EXACT test for the hypothesis of equal proportions.
Proportions of affected pups per litter with necropsy observations: Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians.
Statistics of pathology:
Non-parametric one-way analysis using KRUSKAL-WALLIS test (two-sided). If the resulting pvalue was equal or less than 0.05, a pair wise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the equal medians.
Reproductive indices:
Male and female mating index
Male and female fertility index

gestation index
Offspring viability indices:
Live birth index

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food efficiency:
no effects observed
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
Histopathological findings: neoplastic:
not examined

Reproductive function / performance (P0)

Reproductive performance:
no effects observed

Details on results (P0)

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS): In all male and female animals of test group 3 (1000 mg/kg bw/d) discoloured feces were
observed during premating, gestation and lactation period. No treatment-related findings were observed for all male and female animals of the remaining test groups 1 and 2(100 and 300 mg/kg bw/d). In a single female of test group 0 (0 mg/kg bw/d) no sperm in vaginal smear was detected during the entire mating period, but delivered pups. One female animal of test group 0 (0 mg/kg bw/d) and one female animal of test group 2 (300 mg/kg bw/d) did not deliver. These findings were assessed as being incidental and not related to the test substance administration.
No mortality occurred during the entire study period.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS): No treatment-related changes in body weight data were observed in male and female animals of all test groups. The single significantly increased value in body weight change in females of test group 3 (1000 mg/kg bw/d) from day 0 to 7 during gestation (+24%) was assessed as being incidental.
No significant changes in food consumption were observed for male and female animals during the study period.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS). For all F0 parental males mating was confirmed. Thus, the male mating index was 100%.
The female mating index calculated after the mating period for F1 litter was 100%. The mean duration until sperm was detected (GD 0) amounted to 3.9, 2.6, 1.9 and 3.0 days (0, 100, 300 and 1000 mg/kg bw/d, respectively). The differences between the test groups were assessed as being spontaneous in nature and without biological or toxicological relevance. All animals of all test groups (0, 100, 300 and 1000 mg/kg bw/d) delivered pups or had implantations in utero with exception of one animal of test group 0 and one animal of test group 2. The female fertility index was 90% in test group 0 and 2 and 100% in test group 1 and 3. Thus, no treatment-related effect was observed. The mean duration of gestation was similar in all test groups, i.e. between 22.0 and 22.2 days. Implantation was not affected by the treatment in all test groups since the mean number of implantation sites was comparable among all groups. Postimplantation loss was between 1 and 4, these values reflected the normal range of biological variation inherent in the strain of rats used for this study as all respective values were within the range of the historical control data. The gestation index was 100% in all test groups. The mean number of F1 pups delivered per dam was 11.6, 11.9, 11.9 and 12.2 pups/dam in test groups 0-3 (0, 100, 300 and 1000 mg/kg bw/d, respectively). The rate of liveborn pups was not affected by the test substance as indicated by the live birth indices of 100% in test group 0 and 1. In test group 2 the live birth index was significantly decreased (94%) and in test group 3 slightly decreased (98%). As no dose-response relationship was observed and all respective values were within the range of the historical control data, the lower indices were not assumed to be treatment related.

GROSS PATHOLOGY (PARENTAL ANIMALS): In females of test group 3 (1000 mg/kg bw/d) the contents of the stomach (2/10 females) and rectum (4/10 females) showed an orange discoloration. In a previous study with the same substance (Kaspers, U., et al.) the discoloration of contents occurred without any histopathological correlate. Therefore, stomach and rectum were not examined for histopathology and the discoloration of contents was regarded as non adverse and was considered to respond to the orange color of the test substance.
All other gross lesions observed in the test animals occurred singularly and were considered to be incidental and not related to treatment.

HISTOPATHOLOGY (PARENTAL ANIMALS): One male of the control group showed in the kidneys an unilateral nephroblastoma. This finding, as well as all other histopathological findings were either single observations or biologically equally distributed between control and treatment groups. Such findings were considered to be incidental and/or spontaneous in origin and without any relation to treatment.

Effect levels (P0)

open allclose all
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: reproductive performance and fertility was not affected by the test substance as no dose-response relationship
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no signs on general, systemic toxicity were observed

Target system / organ toxicity (P0)

Critical effects observed:
no

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed

Details on results (F1)

VIABILITY (OFFSPRING): No significant increase of pups that died during lactation was observed for test groups 1-3 (100, 300 and 1000 mg/kg bw/d). The viability index as indicator for pup mortality between PND 0-4 was between 96% and 100% for all test groups.
The mean number of delivered pups per dam and the rate of liveborn pups were evenly distributed among test groups 0-3 (0, 100, 300 and 1000 mg/kg bw/d). The respective values reflect the normal range of biological variation inherent in the strain used in this study.
In test group 2 (300 mg/kg bw/d), the number of liveborn pups was significantly decreased (94%) and the number of stillborn pups was significantly increased. Due to the lack of a dose-response, this was assessed as being incidental.

CLINICAL SIGNS (OFFSPRING): All F1 pups did not show adverse clinical signs up to scheduled sacrifice on PND 4.

BODY WEIGHT (OFFSPRING): Mean pup body weights/pup body weight changes of all pups in test groups 1-3 (100, 300 and 1000 mg/kg bw/d) were comparable to the concurrent control group (0 mg/kg bw/d). The observable differences between the groups were assessed as being spontaneous in nature and without biological relevance.
One runt was seen in female pups of test group 1 (100 mg/kg bw/d). This was within the normal range inherent of this strain and not related to treatment.

SEXUAL MATURATION (OFFSPRING): The sex distribution and sex ratios of live F1 pups on the day of birth and on PND 4 did not show biologically relevant differences between all test groups.

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no developmental toxicity was observed
Remarks on result:
other: Observed up to postnatal day 4

Target system / organ toxicity (F1)

Critical effects observed:
no

Overall reproductive toxicity

Reproductive effects observed:
no

Any other information on results incl. tables

Table 1: Female fertility index

Test groups

Fertility index (%)

0mg/kg bw/d

90

100mg/kg bw/d

100

300mg/kg bw/d

90

1000mg/kg bw/d

100

Applicant's summary and conclusion