Tin sulfide (Sn2S3)

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-12-10 to 2010-06-15

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP and guideline compliant study on a structural analogous read-across substance (Please refer to the Read-Across Justification in Section 13)

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009-12-02 to -2010-06-30

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP and guideline compliant study on a structural analogous read-across substance (Please refer to the Read-Across Justification in Section 13)

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Version / remarks:

, 1995

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Species and strain: Wistar rat
- Supplier: BioTest s.r.o., Czech Republic
- Quality: SPF
- Age on delivery: 6-7 weeks
- Age at treatment start: 7-8 weeks
- Body Weight at delivery: DRF Study: Males 202-227g, Females 169-178g 90-Day Subchronic Study: Males 166-188g, Females 126-160g


- Housing: Macrolon 2000P cages (Tecniplast, 2-3 animals of the same sex/cage)
- Diet: standard pelletized diet NOE H4 (Racio Břeclav, CZ) ad libitum
- Water: drinking water ad libitum
- Acclimatization time: 5 or 6 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2 °C
- Humidity: 30 - 70 %
- Photoperiod: 12 hours daily

Route of administration:

oral: gavage

Vehicle:

other: methylcellulose

Details on oral exposure:

PREPARATION OF THE TEST ITEM FORMULATION:
For the preparation of a homogenous suspension the following procedure was adopted: at first the weighted amount of the respective substance was ground in a grinding mortar with a small volume of carrier liquid. Then the content of the mortar is transferred into a calibrated vessel. Another amount of carrier liquid is poured into the mortar, stirred and the mortar with the pestle is rinsed, the rinsing liquid being added into the same calibrated vessel. This rinsing is repeated once more. Then the volume in the calibrated vessel is made up with the carrier liquid to the required volume and mixed on magnetic stirrer using 1000 rpm for 10 minutes. Immediately before the application, the suspension is again stirred by means of an electromagnetic stirrer.

VEHICLE
methylcellulose is used as vehicle in this study in a concentration of 0.5%

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

90 days

Frequency of treatment:

Three treated groups were administered with doses of 100 mg and 300mg/kg and 1000 mg/kg bw per day. Control group was administered by the vehicle (0.5%) methylcellulose, only.

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

Details on study design:

DRF Study METHOD
Three groups of rats (each group 4 males and 4 females) were daily administered with the Test Item suspension in 0.5% methylcellulose orally by gavage for 14 days. The first two groups were administered with doses of 500 mg/kg (D1 group) and 1000 mg/kg (D2 group). The third group (Control) was administered in the same manner by vehicle (0.5% methylcellulose). All animals were daily checked for clinical signs. Individual body weight and food consumption were recorded weekly. Before the first administration (Day -1) and after the last administration (Day 15) blood and urine samples were taken from all animals for hematology, clinical chemistry and urinalysis examinations. At the end of the study (Day 15) gross necropsy of all rats was performed and gross lesions were recorded.
90-Day Subchronic Toxicity Study METHOD
Four groups of rats (each of 10 males and 10 females) were daily administered with the Test Item suspension in 0.5% methylcellulose orally by gavage for 90 days. Three treated groups were administered with doses of 100 mg/kg (D1 group), 300 mg/kg (D2 group) and 1000 mg/kg body weight/day (D3 group). The fourth group (Control) was administered in the same manner by the vehicle (0.5% methylcellulose). All animals were daily checked for clinical signs. Individual body weight and food consumption were recorded weekly. Before the first administration (Day -1) and at the end of the study (Day 90) blood and urine samples were taken from all animals for hematology, clinical chemistry and urinalysis examinations. Ophthalmoscopy was also performed initially and terminally on all animals. At the end of the study (Day 90), gross necropsy of all rats was performed and organ and tissue samples according to Study plan were taken for histopathological examination.

Positive control:

No

Observations and examinations performed and frequency:

All animals were checked daily for clinical signs. Individual body weight and food consumption were recorded weekly.Before the first administration (day -1) and at the end of the study (day 90) blood and urine samples were taken from all animals for hematology, clinical chemistry and urinalysis examinations. Ophthalmoscopy was also performed initially and termininally on all animals. At the end of the study (day 90), gross necropsy of all rats was performed and organ and tissue samples according to study plan were taken for histopathological examinations.

Sacrifice and pathology:

At the end of the study (day 90), gross necropsy of all rats was performed and organ and tissue samples according to study plan were taken for histopathological examinations.

Other examinations:

Clinical observations, body weight, food consumption, ophthalmoscopy, hematology, clinical chemistry, urinalysis, pathology (organ weights, macroscopy, microscopy)

Statistics:

DRF Study
Data of body weight, food consumption, hematology including bone marrow smears and clinical chemistry as well as gross pathology were not statistically processed in the DRF study. Group mean values and SD values were counted only.
90-Day Subchronic Toxicity Study
Group mean and standard deviations were calculated for body weights, food consumption, clinical pathology (except for urinalysis) values and organ weights (absolute and relative weights). Absolute and relative organ weights for test article treated groups were compared to those of controls (D1, D2 and D3 versus C) using the Student’s t-Test. In addition, body weights, food consumption and clinical pathology values were subjected to analysis of variance (ANOVA) followed by Dunnett’s Test, if applicable; additional statistical analyses (Kruskall-Walis) were performed as required by the data. Statistical evaluation was performed using Graph Pad Prism software, version 4, validated on September 22, 2009.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Tin sulfide caused a slight increase in food consumption, associated with slight increases in body weight and serum glucose concentrations in females dosed at 1000 mg/kg. This was not considered as adverse effect.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Tin sulfide caused a slight increase in food consumption, associated with slight increases in body weight and serum glucose concentrations in females dosed at 1000 mg/kg. This was not considered as adverse effect.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Slight dose dependent decreases in serum Na and Cl, varying within physiological range, were observed. Increased serum glucose concentrations in females dosed at 1000 mg/kg was most probably due increased food consumption and not considered as adverse.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Description (incidence and severity):

No observations indicating neurotoxicity were noted.

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No adverse test item and/or treatment related changes

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

No adverse test item and/or treatment related changes

Details on results:

Tin sulfide orally administered to rats for 90 consecutive days caused a slight increase in food consuption, associated with slight increases in body weight and serum glucose concentrations in females dosed at 1000 mg/kg. These were however not considered as adverse effects. Slight dose dependent decreases in serum Na and Cl, varying within physiological range, were observed. Except for the above mentioned changes, tin sulfide did not cause any negative effect on body weight, food consumption, ophthalmoscopy, hematology and clinical chemistry parameters and organ weights.
Tin sulfide further did not cause any organ weight changes nor gross or histopathological changes in the liver, kidneys, gastrointestinal tract or other organs of survived animals indicative of a toxic effect. Other lesions found in the treated animals were either of spontaneous character or they were not in direct relation with the administration of tin sulfide.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Under the test conditions used, 90 - day administration of the test item tin sulfide to rats was safe and well tolerated up to the dose of 1000 mg/kg bw/day. The NOAEL was defined at 1000mg/kg and the NOEL at 300 mg/kg.

Critical effects observed:

not specified

RESULTS of DRF Study:

All rats survived till their scheduled necropsy. No signs of toxicity were recorded in all administered animals of all tested groups during the observation period. Variability observed in body weight and food consumption were not treatment related and both recorded parameters were not influenced with the test item administration. No treatment-related changes were found in the red blood cells, white blood cells and coagulation parameters. The decrease in APTT values in dose group D1 was probably no treatment related. No treatment related changes were observed during the study in the clinical chemistry and urinalysis parameters monitored. No gross pathology findings were found during necropsy of the animals. In conclusion, 14 days oral administration of tin sulfide at doses of 500 and 1000 mg/kg body weight/day in rats was safe and well tolerated. Based on the obtained results it was proposed to design the 90-day Subchronic Toxicity Study with doses of 100, 300 and 1000 mg/kg body weight/day.

Conclusions:

Tin sulfide showed no effects considered as adverse in a subchronic toxicity study according to OECD guideline 409. The NOAEL was defined at 1000 mg/kg and the NOEL at 300 mg/kg.

Executive summary:

A 90-day study according to OECD guideline 409 with tin sulfide was performed in rats to provide information on the possible health hazards likely to arise from repeated oral exposure over a prolonged period of time covering post-weaning maturation and growth well into adulthood. Three treated groups were administered with doses of 100 mg and 300mg/kg and 1000 mg/kg bw per day. Tin sulfide caused a slight increase in food consumption, associated with slight increases in body weight and serum glucose concentrations in females dosed at 1000 mg/kg. These were however not considered as adverse effects. Slight dose dependent decreases in serum Na and Cl, varying within physiological range, were observed. Except for the above mentioned changes, tin sulfide did not cause any negative effect on body weight, food consumption, ophthalmoscopy, hematology and clinical chemistry parameters and organ weights. Tin sulfide further did not cause any organ weight changes nor gross or histopathological changes in the liver, kidneys, gastrointestinal tract or other organs of survived animals indicate of a toxic effect. Under the test conditions used, 90-day administration of the test item to rats was safe and well tolerated up to the dose of 1000 mg/kg bw/day. The NOAEL was defined at 1000mg/kg and the NOEL at 300 mg/kg.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)

Version / remarks:

adopted by the Council on July 27th 1995

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Tin sulphide

EC Number:

215-248-7

EC Name:

Tin sulphide

Cas Number:

1314-95-0

Molecular formula:

SSn

IUPAC Name:

Tin(II) sulfide

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Species and strain: Albino laboratory rat Wistar Han ( outbred SPF quality - guaranteed)
- Supplier: SPF breeding, VELAZ s.r.o., Kolec u Kladna Czech Republic, RCH CZ 21760152
- Sex: males and virgin females
- Age at study initiation: 10 weeks
- total numer of animals: 48 males and 48 females (12 females and 12 males per group)
- Housing: Animals were housed in SPF animal room, 2 rats of the same sex in one plastic cage ( 40x25x20 cm) containing sterilised clean shavings of soft wood. During mating period - one male and one female in one cage, pregnant females -individually, offspring- with mother.
- Diet: Complete pelleted diet for rats and mice in SPF breeding- ST 1 BERGMAN was used, manufacturer: Mlyn Kocanda, V)'roba krmnych smesf, Kocanda No.19, 252 42 Jesenice u Prahy. Diet was sterilised before using .
- Water: Free access to drinking water (ad libitum). Water quality corresponded to Regulation No. 252/2004 Czech Coil. of Law, Health Ministry. Water was sterilised before using.
- Acclimatization time: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19.9 to 23.2°C
- Humidity: 35.8 to 66.6 %
- Air changes: approximately 15 air exchanges/hour
- Photoperiod: 12 hours daily

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% methylcellulose in water for injection

Details on exposure:

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 substance 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).
The test substance was administered to the stomach by gavage as suspension in 0.5% methylcellulose in water for injection.Oral administration of rats was made daily.

Details on mating procedure:

Animals were mated from the 15th day of study. Mating 1: 1 ( one male to one female) was used in this study. Each morning the females were examined for presence of spermatozoa in vaginal smears. Day 0 of pregnancy was defined as the day the sperms were found.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analytical report includes the results of testing a homogeneity of Tin sulfide application form.
The application form for "Tin sulfide- Reproduction/Developmental Toxicity Screening Test" is the suspension of test substance in 0.5% methylcellulose. The test purpose was found out whether the application form prepared is homogenous.
Homogeneity of the application form was checked by determination of a concentration of the test substance in three levels of solution (at the bottom, in the middle and at the surface ).
The measurement was performed on two concentration levels (50 mg / 10 mL and 1000 mg / 10 mL). The application form was prepared in a way that is conformable with "Tin sulfide – Reproduction/Developmental Toxicity Screening Test" (the similar equipment: container, mixing stirrer). The analyses were performed by ICP - OE spectrophotometric method.
From the results of analyses follows that the Tin sulfide application form in vehicle at defined laboratory conditions (laboratory temperature, preparation of solution by defined manner) is homogeneous. The differences in concentrations detected on the single levels of measurement to average are less then uncertainty of measurement.

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:

once per day, 7 days per week

Details on study schedule:

- Age at mating of the mated animals in the study: 12 weeks (10 weeks at start of study; mating from the 15th day of study)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

300 mg/kg bw/day (actual dose received)

Doses / concentrations 3

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:
The dose levels for study- 100, 300 and 1000 mg/kg/day -were chosen with respect to the results of the dose-range finding experiment and on request of the sponsor.
The DRF experiment with 14-day application period was performed with 4 groups of treated rats. The doses for the DRF experiment were chosen with respect to the literature data : 125, 250, 500 and 1000 mg/kg/day. In the DRF experiment with Tin sulfide, the body weight increments and haematological parameters were relatively well-balanced at all dose levels during the 14-day application period. The pathological examination did not reveal any pathological changes. No animal died during the 14-day application period.
- Rationale for animal assignment (if not random): random

Positive control:

no

Examinations

Parental animals: Observations and examinations:

Health condition control: daily - during the acclimatization and the experimental part;
Body weight: males - the first day of administration and then weekly,females - the first day of administration and then weekly in prematingand mating period; during pregnancy: 0., 7th, 14th, 20th day; during lactation: 0. or 1st and 4th day;
Food consumption: males- weekly (except the mating period), females - weekly during premating period and after mating period, during pregnancy and lactation on the same days as body weight;
Clinical observations: males and females - daily during the administration period;
Mortality control: daily.

Oestrous cyclicity (parental animals):

The oestrous cycle length and pattern was not evaluated before mating but the phases of oestrous cycle were recorded during histopathological examination.

Sperm parameters (parental animals):

In all males of all groups surviving to scheduled necropsy the sperm parameters were examined: sperm motility and sperm morphology. Sperm specimens were prepared and examined according to the SOP No. M/45.
Sperm motility:
Sperm samples were taken from one epididymis and sperm motility was assessed from these samples. The motility of sperm was determined by microscopic examination of the prepared sperm suspension (mixture of one epididymis cutted into pieces and 5 ml of 0.9% NaCl). The result of observation was evaluated subjectively according to following grades: 1 - fast progressive motility, 2 - slow progressive motility, 3 -no progressive motility, 4- nonmotile sperm.
Sperm morphology:
Sperm samples (taken from one epididymis) and sperm morphology was assessed from these samples. A smear from the sperm suspension was prepared and stained (Giemsa staining). The morphology of sperm was determined by microscopic examination. All deviations - e.g. broken tail, abnormal form of tail, double head, amorphous head and abnormal form of neck - were recorded.

Litter observations:

All pups were observed in natural conditions in their cages daily during the lactation period. Changes in behavioural abnom1alities were recorded. Detailed examination of each litter was performed as soon as possible after delivery (day 0 or 1 post-partum) and the 4th day of lactation. The number and sex of pups, stillbirths, live births and presence of gross anomalies were recorded.

Postmortem examinations (parental animals):

Parental males were killed at the end of the administration period after 42 days of administration. Parental females were killed on the 4th day of lactation. Mated females without delivery was killed 26th day after confirmed mating. Then they were macroscopically examined for any pathological changes with special attention to the organs of the reproductive systems. All macroscopic abnormalities were recorded. The absolute weights of testes, one epididymis, prostate gland and pituitary gland were recorded in males and absolute weight of ovaries; uterus (incl. uterine tube and cervix) and pituitary gland were recorded in females. Afterwards the relative weight of the organ was computed according to the following formula: weight of organ x 100/ body weight.

Postmortem examinations (offspring):

Dead pups were sexed and externally examined; the stomach was examined for the presence of milk. Pups killed on the 4th day of lactation were sexed and subjected to external examination of the cranium, and to macroscopic examination of the thoracic and abdominal tissues and organs. All macroscopic changes were recorded.

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 0/1 st day and 4th day of lactation period and average weight of pup on the 0/1 st 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).

Reproductive indices:

Reproduction parameters: number of females achieving pregnancy, number of females bearing live pups and number of females with live pups at day 4 after parturition in treated groups were similar to the control or higher than the control groups. Duration of mating and pregnancy were similar in the control and treated females. Pre-implantation, post-implantation and postnatal losses were relatively well balanced at the treated groups and control group.

Offspring viability indices:

Pre-implantation loss, post-implantation loss, post-natal loss, mating index, fertility index, conception index, gestation index, percentage of postnatal loss days 0-4 post partum and viability index were calculated.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Males:
No treatment-related effects were found in treated males during the health condition control.
No relevant clinical changes were observed in males at all dose levels after application of the test substance.
Slight fall of weight ( one male at the dose level of 100 mg/kg/day in the 6th week) and thinner excrements ( one male at the dose of 1000 mg/kg/day in the 1 st week) were observed sporadically in treated males.
Females:
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 during 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):

Males: Average food consumption values of males at all treated close levels were slightly increased against the control group during the whole study. These differences were not dependent on the dose level and therefore not considered relevant.
Fermales:
Females:
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 well-balanced 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:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

The effect on the microscopical structure of the testes (sporadic degeneration and/or atrophy of germ epithelium, residual bodies in germ epithelium and vacuolation of cytoplasm of spermiogonia) had no effect on spermiogenesis, thus, were not relevant.

Histopathological findings: 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

Details on results (P0)

Body weight, food consumption, clinical status and macroscopic structure of reproductive organs of treated parental males were unaffected by treatment of the test substance. A statistically significant difference was detected in the weight of the pituitary gland. An absolute weight of pituitary gland was statistically significantly incrased in males at the dose level of 1000 mg/kg/day. In absence of any changes of microscopic structure of the pituitary gland, the effect could be considered not to be of toxicological importance. The test substance had effect on the microscopical structure of the tests without damage of spermiogenesis. Histopathological examination of testes of parental males showed increased incidence of irrelevant changes: degenerations and/or atrophies of germ epithelium and vacuolations of cytoplasm of spermiogonia in males of the dose level 1000 mg/kg/ day. However, there were no histopathological findings in testes in a 90-day study on tin sulfide. Food consumption, clinical observations, weight and structure of reproductive organs of treated parental females were not significantly affected by treatment of the test substance. Growth of parental females was slightly but insignificantly influenced by the test substance administration: the body weight of females at the dose levels of 300 and 1000 mg/kg/day were slightly decreased during the pregnancy and lactation period.
Reproduction parameters - number of females achieving pregnancy, number of females bearing live pups and number of females with live pups at day 4 after parturition in treated groups were similar to the control or higher than the control groups. Duration of mating and pregnancy were similar in the control and treated females. Pre-implantation, post-implantation and postnatal losses were relatively well balanced at the treated groups and control group.

Effect levels (P0)

1

Key result

Dose descriptor:

NOAEL

Remarks:

for the REPRODUCTION

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Systemic toxicity

2

Key result

Dose descriptor:

NOEL

Remarks:

toxic effect on reproduction organs

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Sex:

male

Basis for effect level:

organ weights and organ / body weight ratios

histopathology: non-neoplastic

3

Key result

Dose descriptor:

NOEL

Remarks:

toxic effect on reproduction organs

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

reproductive performance

Results: F1 generation

General toxicity (F1)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Description (incidence and severity):

One pup of control group and one pup of the dose level 300 mg/kg/day died after birth – in the period 0/1st day - 4th day.
No differences in development of pups were observed at the control group and at all treated groups.
Statistical evaluation was performed on the number of live pups.
The total numbers of live pups (on the day of parturition/1st 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 dose 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 historical control data.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The average weights of the litters and pups at the dose level 100 mg/kg/day were slightly increased against control. At the dose 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. No statistically significant changes were found.

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Urinalysis findings:

not examined

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Description (incidence and severity):

The macroscopical examination was performed in all pups. Sporadic pathologic findings were recorded in pups of the treated and control groups. The number of pups with macroscopic changes was comparable in the control and 100 and 300 mg/kg dosed groups; an increase in livers with marked structure was noted in the 1000 mg/kg dose group.

Histopathological findings:

no effects observed

Description (incidence and severity):

The microscopical examination was performed in the pups with macroscopic changes: susp. hydrocephalus (litter No. 142 at the dose level 300) - no histopathological changes in cranial cavity; dark spot on testis (litter No. 167 at the dose level 1000) - no histopathological changes on testis and epididymis; oversized teeth (litter No. 127 at the dose level 100) – no histopathological changes and marked structure of liver (litter No. 164 and 171 at the dose level 1000) – extramedullary haemopoiesis was recorded (it is a physiological finding in pups).

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Details on results (F1)

The number of corpora lutea, implantations, number of pups and accompanied weight of litter were slightly but insignificantly influenced by the administration of the test substance (decrease esp. at dose levels 300 and 1000 mg/kg/day). Sex ratio, average weight of pups and postnatal development of pups were unaffected. Macroscopic abnormalities were described sporadically in pups of all treated and the control groups.

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Remarks:

for the development

Generation:

F1

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

viability

clinical signs

mortality

body weight and weight gain

gross pathology

Overall reproductive toxicity

Reproductive effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

A reproduction and development toxicity screening study according to OECD guideline 421 in rats with tin sulfide revealed the following values:
NOAEL for the Reproduction: 1000 mg/kg bw/day,
NOEL for the toxic effect on reproduction organs of males: 300 mg/kg bw/day,
NOEL for the toxic effect on reproduction organs of females: 1000 mg/kg bw/day,
NOAEL for the Development of pups: 1000 mg/kg bw/day.

Executive summary:

A reproduction and development toxicity screening study according to OECD guideline 421 in rats with tin sulfide revealed the following values:

NOAEL for the Reproduction: 1000 mg/kg bw/day,

NOEL for the toxic effect on reproduction organs of males: 300 mg/kg bw/day,

NOEL for the toxic effect on reproduction organs of females: 1000 mg/kg bw/day,

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

The irrelevant negative influence of the test substance treatment expressed in males of the highest dose level (limit dose) consisted of an increase in absolute weight of pituitary gland (without histopathological changes) and an effect on the microscopical structure of the testes (sporadic occurrence of degeneration and/or atrophy of germ epithelium, residual bodies in germ epithelium and vacuolation of cytoplasm of spermiogonia) without effect on the spermiogenesis. Moreover, there were no histopathological findings in testes in a 90-day study on tin sulfide. The average number of pups and accompanied weight of the litters varied among groups, but fell withing historical ranges and these differences were not considered to be toxicologically relevant.

Body weight, food consumption, clinical observations, weight of reproductive organs of parental males were not markedly affected by treatment of the test substance. Body weight, food consumption, clinical observations, organ weights and structure of reproductive organs of parental females were also not adversely affected by treatment of the test substance. Reproductive performance - ability of male and female animals to successfully mate and produce viable offspring was unaffected by the test substance treatment. Also sex ratio and development of pups were not changed in treated groups.