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Toxicological information

Developmental toxicity / teratogenicity

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Administrative data

Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2009-12-10 to 2012-06-15
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was conducted with a similar substance according to standard methods and GLP, therefore it is considered reliable and relevant, but as it is only a screening study, not adequate for classification. Various anchor points demonstrated a similar structural and physicochemical characteristics and comparable toxicological profile ( acute toxicity, genotoxicity and local tolerance endpoints), showing that read-across can be considered adequate.
Cross-reference
Reason / purpose:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: OECD 421
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): Tin Sulfide
- Molecular formula (if other than submission substance): SSn
- Molecular weight (if other than submission substance): 150.776
- Smiles notation (if other than submission substance): [SnH2]=S
- InChl (if other than submission substance): 1S/S.Sn
- Substance type: Inorganic monoconstituent
- Physical state: Dark grey powder
- Analytical purity: ca. 97.6% (77.5% Sn, 20.1% S)
- Lot/batch No.: CH S80774
- Expiration date of the lot/batch: Till 08/2011
- Stability under test conditions: In the time period used for the handling with suspension in this study, Tin sulfide was stable and not decomposed. ( The application form of Tin sulfide was prepared daily in the morning and it was immediately administered to animals.)
- Storage condition of test material: In supplied glass container in the dark and in dry room, at the ambient temperature

Test animals

Species:
rat
Strain:
Wistar
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: SPF Breeding, VELAZ s.r.o., Koleč
- Age at study initiation: 10 weeks
- Weight at study initiation: Males average 317.67 g; Females average 212.17 g
- Fasting period before study: Not provided
- Housing: In SPF animal room, 2 rats of the same sex in one plastic cage (40x25x20 cm) containing sterilized clean shavings of soft wood. During mating period-one male and one female in one cage, pregnant females-individually, offspring-with mother.
- Diet (e.g. ad libitum): Complete pelleted diet for rats and mice in SPF breeding –ST 1 BERGMAN (Mlýn Kocanda, Výroba krmných smesi, Kocanda No. 19, 252 42 Jesenice u Prahy. Diet was sterilized before using.
- Water (e.g. ad libitum): Water was sterilized before using, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%): 30-70%
- Air changes (per hr): 15/h
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5% methylcellulose in water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The concentrations of the suspensions at all dose levels were adjusted to ensure the administration of 1 mL per 100 g of body weight. The vehicle control group was administered by 0.5% methylcellulose in water for injection in the same volume. The application form (test susbstance suspension in 0.5% methylcellulose in water for injection) was prepared daily just before administration. This suspension was mixed for 10 minutes by magnetic stirrer (ca. 1000 rpm).


VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle:
- Amount of vehicle (if gavage):
- Lot/batch no. (if required): methylcellulose: DT223712; water for injection: 0204101109, 0101011209
- Purity:
Analytical verification of doses or concentrations:
yes
Details on mating procedure:
- Impregnation procedure: cohoused
- If cohoused:
- M/F ratio per cage: 1/1
- Length of cohabitation: until day 0 of pregnancy
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:
The treated groups were administered daily for the following periods:
-males and females : 2 weeks prior to the mating period and during the mating period
-pregnant females: during pregnancy and till the 3rd day of lactation
-males: after mating period; totally for 42 days
-non-pregnant females (mated females without parturition): for 25 days after the confirmed mating
Frequency of treatment:
daily
Duration of test:
at least 4 weeks
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:
12
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: dose range finding study (Annex 2) and request of the sponsor

Examinations

Maternal examinations:
CAGE SIDE OBSERVATIONS: Yes : health condition control
- Time schedule: daily during the acclimatization and the experimental part

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily during the administration period; mortality control: daily

BODY WEIGHT: Yes
- Time schedule for examinations: the first day of administration and then weekly in premating and mating period; during pregnancy: 0., 7th, 14th, 20th day; during lactation: 0. or 1st and 4th day

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes weekly during premating period and after mating period; during pregnancy and lactation –on the same days as body weight
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes : in the pregnancy and lactation period (for each week)

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on the 4th day of lactation
- Organs examined: relevant gross lesions, pituitary gland, coagulation gland, cervix of uterus, ovaries, uterus and vagina; absolute weight of ovaries, uterus (incl. uterine tube and cervix) and pituitary gland; afterwards the relative organ weight was computed (weight of organ x 100/ body weight)

OTHER:
- vaginal smears were prepared daily during the mating period (until the presence of spermatozoa)
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: No (uterus weight incl. uterine tube and cervix)
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes (number of corpora lutea – number of implantations)
- Number of late resorptions: Yes (number of implantations – number of live births)
- Other: Post-natal loss (number of live births - number of alive at postnatal day 4)
Fetal examinations:
- External examinations: Yes (Pups killed on the 4th day of lactation were subjected to external examination of the cranium, and to macroscopic examination of the thoracic and abdominal tissues and organs)
- Soft tissue examinations: No
- Skeletal examinations: No
- Head examinations: No

Statistics:
The ANOVA test –Analysis of Variance (QC. Expert 2.5) at significance level 0.05 was used for the statistical analysis. This statistical analysis was used for the results of body weight –necropsy weight in males, body weight of females on the 20th day of pregnancy, necropsy weight of females; biometry of organs of males and females; number of live pups on the 0/1st day and 4th day of lactation period and average weight of pup on the 0/1st day and 4th day of lactation period. Control group with vehicle was compared with the three treated groups.
The results statistically significant on probability level 0.05 are indicated by figures with asterisk in the summary tables (no findings at probability level 0.01 and 0.001).

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Slight decreases in body weights were sporadically (in 1 or 2 females from the group) recorded in the 3rd week at all groups including control.
In treated females of all dose levels, no signs of disease were found during the check-in, acclimatisation and application period. Treatment-related effects were not detected during health condition control and clinical observation of females ( except of vocalisation in one female at the dose level of 1000 mg/kg/day in the 5th week).
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths during the study.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
The average animal body weights at all dose levels were relatively balanced with the control group d
uring the whole application period. No statistically significant differences were detected.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Pre-mating period: Average food consumption of treated females was analogous to food consumption of control females.
Pregnancy: Females without parturition (non pregnant or aborted females) were not included in the evaluation of food consumption during pregnancy. Average food consumption of treated females was wellbalanced with control females.
Lactation: Only mothers (females with live pups) were included in evaluation of food consumption during lactation period. Average food consumption of treated females was similar to consumption of control females.
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
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The incidence of affected females is expressed in numeric form and ranged in sequence of the dose levels of 0-100-300-1000 mg/kg/day further in the text. Examination of the external surface of the body revealed no changes. In the thoracic and abdominal cavities, no changes were observed. In the pelvic area, dilatation of uterine horns by clear liquid was detected in 3-1-1-0 females. No other macroscopic findings were recorded.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed

Maternal developmental toxicity

Number of abortions:
effects observed, non-treatment-related
Description (incidence and severity):
Only Female No. 128 aborted (during necropsy implantations in uterus were found); as this was a low dosed animal, the finding was not related to treatment.
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
No significant differences in pre-implantation and postimplantation llosses were detected between the control and treated mothers.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
No resorptions were reported.
Early or late resorptions:
no effects observed
Dead fetuses:
no effects observed
Changes in pregnancy duration:
no effects observed
Description (incidence and severity):
Duration of mating and pregnancy of treated groups was similar to the control group.

Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.DescriptionIncidenceAndSeverityEffectsOnPregnancyDuration): Duration of mating and pregnancy of treated groups was similar to the control group.
Changes in number of pregnant:
no effects observed
Description (incidence and severity):
The number of females achieving pregnancy and the accompanying conception index were comparable between the control and 300 mg/kg/day dose level. At the dose levels of 100 and 1000 mg/kg/day, the numbers of females achieving pregnancy and accompanying conception indexes were increased compared to the control group.
Other effects:
no effects observed
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
See section 7.8.1

Effect levels (maternal animals)

open allclose all
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: developmental toxicity
Dose descriptor:
NOEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Basis for effect level:
other: maternal toxicity

Results (fetuses)

Fetal body weight changes:
no effects observed
Description (incidence and severity):
The average weight of pups was slightly higher in treated groups against the control group. All values however fell within historical range, therefore this variation was considered to be coincidental.
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): no effects observed
Field "Description (incidence and severity)" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.DescriptionIncidenceAndSeverityFetalPupBodyWeightChanges): The average weight of pups was slightly higher in treated groups against the control group. All values however fell within historical range, therefore this variation was considered to be coincidental.
Reduction in number of live offspring:
effects observed, treatment-related
Description (incidence and severity):
A slightly lower average number of pups per mother accompanied by lower weight of the litter was detected at the dose levels of 300 and 1000 rng/kg/day but the numbers were not out of historic control.
Changes in sex ratio:
no effects observed
Description (incidence and severity):
Sex ratio of pups was balanced in treated and control groups.
Changes in litter size and weights:
no effects observed
Description (incidence and severity):
A slightly lower average number of pups per mother accompanied by lower weight of the litter was detected at the dose levels of 300 and 1000 mg/kg/day but the numbers were not out of historic control.
Changes in postnatal survival:
no effects observed
Description (incidence and severity):
The number of pups on the 4th day of lactation were insignificantly influenced by the test substance treatment.
External malformations:
no effects observed
Description (incidence and severity):
Macroscopie abnormalities were only sporadically detected at necropsy of pups and microscopical evaluation of these findings did not reveal relevant pathological effects.
Skeletal malformations:
not examined
Visceral malformations:
not examined
Other effects:
no effects observed
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
The total numbers of live pups (on the day of parturition/1 st day after parturition and the 4th day after parturition) were similar at the control and the dose level 1000 mg/kg. At the dose level 100 mg/kg/day the number was higher, and vice versa at the dose level 300 mg/kg/day, it was slightly lower. Average numbers of pups per litter were well-balanced at the control and the close level 100 mg/kg/day. At the dose levels 300 and 1000 mg/kg/day, the average number of pups per litter was decreased compared to the control, however they fell within the normal range of historica! control data (see Annex 3). Sex ratio of pups was balanced in treated and control groups.
The average weights of the litters and pups at the close level 100 mg/kg/day were slightly increased against controL At the close levels of 300 and 1000 mg/kg/day, the average weight of the litter was lower compared to control, but the average body weights of pups were slightly higher compared to control group.
One pup of control group and one pup of the close level 300 mg/kg/day died after birth - in the period 0/1 st day - 4th day. No differences in development of pups were observed at the control group and at all treated groups. No statistically significant changes were found.

Effect levels (fetuses)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: no effects

Fetal abnormalities

Abnormalities:
not specified

Overall developmental toxicity

Developmental effects observed:
not specified

Any other information on results incl. tables

See section 7.8.1 (Toxicity to reproduction):

Table 4. Number of live pups and sex

Table 5. Average body weight-Pups

Table 6. Reproduction data

Table 7. Fertility parameters

Applicant's summary and conclusion

Conclusions:
NOAEL for the Development of pups: 1000 mg/kg bw/day.

Referring to the results of OECD 421 test, there is no evidence that the test item Tin sulfide is causing any developmental as well as teratogenetical toxicity. After detailed checking "pubmed" source and other sources, even there is no evidence found in epidemiological studies.
Executive summary:

The OECD 421 Screening Test can be used to provide initial information on possible effects on reproduction and/or development, either at an early stage of assessing the toxicological properties of chemicals, or on chemicals of concern. It can also be used as part of a set of initial screening tests for existing chemicals for which little or no toxicological information is available, as a dose range finding study for more extensive reproduction/developmental studies, or when otherwise considered relevant. Males were dosed for a minimum of four weeks and up to and including the day before scheduled kill (this includes a minimum of two weeks prior to mating, during the mating period and, approximately, two weeks post-mating). Females were dosed for at least 2 weeks prior to mating, during mating and gestation up to the third day of lactation.

For the reproductive findings, reference is made to Section7.8.1. For the developmental part of the study, the number of pups, average weight of litter, average body weight of pups on the day of parturition, 1st day after parturition and the 4th day of lactation were insignificantly influenced by the test substance treatment. A slightly lower average number of pups per mother accompanied by lower weight of the litter was detected at the dose levels of 300 and 1000 mg/kg/day but the numbers were not out of historic control. Vice versa, the average weight of pups was higher in treated groups against the control group. Development of pups was not affected by the test substance treatment. Macroscopic abnormalities were only sporadically detected at necropsy of pups and microscopical evaluation of these findings did not reveal relevant pathological effects. In conclusion, NOAEL for the Development of pups was 1000 mg/kg bw/day.