Reference type

Author

Year

Title

Bibliographic source

Testing laboratory

Report no.

Owner company

Company study no.

Report date

study report

MHLW, Japan

2009

A combined repeated-dose/reproductive-developmental toxicity study of Benzenesulfonic acid, 4-hydroxy-, tin(2+)salt by oral administration in rats.

Japan Existing Chemical Data Base (JECDB)

BoZo Research Center Inc

R-945

2009-04-10

data published

7.8.1 Toxicity to reproduction: Toxicity to reproduction.001

subacute

no

Qualifier

Guideline

Deviations

according to

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

no

yes

yes

Identifier

Identity

CAS number

70974-33-3

- Name of test material (as cited in study report): Benzenesulfonic acid, 4-hydroxy-, tin(2+)salt
- Lot No.: 060501
- Supplier: Daiwakasei Industry Co., Ltd
- Chemical structure: [C6H4(OH)SO3]2Sn
- Chemical formula: C12H10O8S2Sn
- Molecular weight: 465.05
- Puitality: 96.0%
- Impurities: Benzenesulfonic acid, 2-hydroxy-, tin(2+)salt; <2% ; Benzenesulfonic acid, 2,4-hydroxy-, tin(2+)salt; <2%
- Boiling point: 300°C or more
- Solubility: White turbidity in 2% ethanol
- Storage condition of test material: Cold dark place at 2 - 8°C

rat

Crj: CD(SD)

male/female

TEST ANIMALS
- Source: Atsugi Breeding Center, Charles River Laboratories Japan, Inc.
- Age at study initiation:10 weeks of age
- Weight at study initiation: 340-409 g for males and 205-257 g for females
- Housing: bracket-type metallic wire-mesh cages (W 250 × D 350 × H 200 mm)
- Diet (e.g. ad libitum):ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period:15 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C):21 to 26°C
- Humidity (%): 38 to 63%
- Air changes (per hr):10 to 15 times per hour
- Photoperiod (hrs dark / hrs light):12-hour lighting per day

oral: gavage

water

PREPARATION OF DOSING SOLUTIONS: Test substance was dissolved in distilled water for injection.
VEHICLE
- Justification for use and choice of vehicle: No data
- Amount of vehicle (if gavage): 5 mL/kg bw
- Lot/batch no. (if required): No data
- Dosing volume: 5 mL/kg
- Storage condition of test solution: Stored in a refrigerator

yes

Test suspensions at each concentration to be used for males in week 1 and 6 of administration were analyzed by the HPLC method at Bozo Research Center Inc. Results showed that the concentration of the test article in each suspension was 94.5 to 107.5% of the nominal concentration, and both values were within the acceptable range (concentration: percentage of the nominal concentration, 100 ± 10%; C.V.: 10% or below)

All males and females without mating: 42 days
Females with mating: up to 51 days including 14 days pre-mating, mating and gestation periods and until day 4 of lactation

Daily: 7 times / week

0 (vehicle), 12, 60 and 300 mg/kg bw/day

Basis

actual ingested

12 animals/sex/dose with mating (main dose group), 5 animals/sex/dose at 0 and 300 mg/kg bw/day without mating (recovery group)

yes, concurrent vehicle

- Dose selection rationale: A preliminary study was conducted to determine the doses to be employed. Male and female rats were receiving 0, 100, 300, and 1000 mg/kg bw/day (5 animals/sex/dose) of the substance was administered for 14 days. As the results, thickening in limiting ridge of the stomach was observed in 2 males and 2 females receiving 100 mg/kg bw/day. Thickening in limiting ridge of the stomach and dilatation in the cecum were observed in almost all animals receiving 300 mg/kg bw/day or more. Decreases in body weights in males and food consumption in both sexes, and increases in AST and ALT activities were observed in an early administration period at 1000 mg/kg bw/day. Therefore, the high dose was set at 300 mg/kg/day, and the middle and low dose were set at 60 and 12 mg/kg/day using common ratio 5.

CAGE SIDE OBSERVATIONS: Yes
- Time schedule:
Males and females: once a day before the start of administration, 3 times/day during the administration period, and once a day during the recovery period
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule:
Males: once before the start of administration, once a week during the administration and recovery periods
Females: once before the start of administration, days 1, 7, 14 and 20 of gestation, and day 4 of lactation
BODY WEIGHT: Yes
- Time schedule for examinations:
Males in the main groups were weighed on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 of administration and on the day of necropsy, and males and females in the recovery groups were weighed on days 1, 4, 8, 11 and 14 of recovery and on the day of necropsy.
Females in the main groups were weighed on days 1, 4, 8, 11 and 15 of administration (uncopulated animals were weighed on day 18 and 22 of administration as well), days 0, 4, 7, 11, 14, 17 and 20 of gestation, days 0 and 4 of lactation and the day of necropsy.
FOOD CONSUMPTION : Yes
Measurement of food consumption was conducted on all animals at the following frequencies:
males in the main groups: on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration; males and females in the recovery groups on days 1, 4, 8, 11 and 14 of recovery in addition to the measurement days for males in the main groups; and females in the main groups on days 1, 4, 8, 11 and 15 of administration, days 1, 4, 7, 11, 14, 17 and 20 of gestation and days 2 and 4 of lactation.
FOOD INTAKE: No
HAEMATOLOGY: Yes
- Time schedule for collection of blood: On the next day of the last administration and on the last day of the recovery period
- Anaesthetic used for blood collection: Yes (identity)
- Animals fasted: Yes
- How many animals:5 animals/sex/group
- Parameters examined: red blood cell count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, reticulocyte percentage, platelet count, white blood cell count, differential white blood cell count, absolute number of each white blood cell, prothrombin time, activated partial thromboplastin time, fibrinogen volume
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: On the next day of the last administration and on the last day of the recovery period
- Animals fasted: Yes
- How many animals: 5 animals/sex/group
- Parameters checked: ALP, total cholesterol, triglyceride, phospholipids, total bilirubin, glucose, blood urea nitrogen, creatinine, sodium, potassium, chloride, calcium, inorganic phosphorus, total protein, albumin, A/G ratio, AST (GOT), ALT (GPT), LDH, γ-GTP
URINALYSIS: Yes
- Time schedule for collection of urine (males): final week of administration period (days 37 to 38 of administration) and in the final week of recovery period (days 9 to 10 of recovery)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
Collection of four-hour urine samples under fasting but ad libitum drinking conditions, followed by collection of 20-hour urine samples under ad libitum feeding and drinking conditions.
- Parameters checked: pH, protein, ketones. glucose, occult blood, bilirubin, urobilinogen, color, sediment, urine volume (4-hour sample), osmotic pressure, urine volume (20-hour sample), water intake (4-hour+20-hour volume)
BLOOD HORMONE: No
- Time schedule for collection of serum: No
- Parameters checked: No
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Manipulative test and measurements of grip strength and motor activity were conducted on 5 animals per group with the following frequencies: males in the main groups were examined in the final week of administration (day 36 of administration), females in the main groups on day 4 of lactation (day 42 and 43 of administration) after necropsy of F1 pups, and males and females in the recovery groups in the final week of administration (day 36 of administration) and in the final week of recovery (day 8 of recovery).
- Dose groups that were examined: All animals were examined for detailed clinical signs once before the start of administration. Thereafter, males in the main groups were examined once weekly during the administration period, whereas females were observed once weekly during the pre-mating administration period and mating period as well as on designated days during the gestation and lactation periods (days 1, 7, 14 and 20 of gestation, and day 4 of lactation). Animals in the recovery groups were examined once weekly during the administration and recovery periods.
- Battery of functions tested:
1) Open field observation. Arousal, gait, posture, tremor, convulsion, rearing count, defecation (defecation count, urination), stereotypy (grooming, circling, etc.), abnormal behavior (self-biting, backward walking, etc.)
2) Manipulative Test. Auditory response, approach response, touch response, tail pinch response, pupillary reflex, aerial righting reflex, landing foot splay
3) Measurement of Grip Strength. Following manipulative test, grip strength of forelimb and hind limb was measured by CPU gauge MODEL-9502A (AIKOH Engineering Co., Ltd.).
4) Measurement of Motor Activity.Following measurement of grip strength, motor activity was measured by a motor activity sensor for experimental animals NS-AS01 (NeuroScience, Inc). The measurement was conducted for 1 hour, and measured values at 10-minute intervals and from 0 to 60 minutes were recorded.

GROSS PATHOLOGY AND ORGAN WEIGHTS：Yes Brain, thyroids(including parathyroids), thymus, heart, liver, spleen, kidneys, adrenals, testes, and epididymis
HISTOPATHOLOGY: Yes Cerebrum, cerebellum, pituitary, spinal cord (thoracic), thyroid, parathyroid, adrenal glands, thymus, spleen, submandibular lymph nodes, mesenteric lymph nodes, heart, lung (including bronchial), stomach, duodenum, jejunum, ileum, cecum, colon, rectum, liver, kidney, bladder, testis, epididymis, ovary, uterus, seminal vesicles, sternum and femur (including bone marrows), macroscopic lesions, and parts for identification (auricles)

The data were analyzed for homogeneity of variance by the Bartlett test (level of significance: 0.01, two-tailed). If variances were homogeneous, data were analyzed by the Dunnett test, whereas heterogeneous data were analyzed by the Steel test. In the recovery test, these values of two groups were analyzed by F test and Student or Aspin-Welch t-test. Frequency data were analysed by Fisher test. Statistical significance was set at < 5% by two-sided

Endpoint

Effect level

Based on

Sex

Basis for effect level / Remarks

NOAEL

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

act. ingr.

male/female

Decreases in hemoglobin and hematocrit levels(females), increases in ALT (males), and lesions in the caecum, stomach and duodenum at 300 mg/kg bw/day.

yes

no effects

no effects

not examined

yes

yes

no effects

no effects

no effects

yes

yes

no data

CLINICAL SIGNS AND MORTALITY: Mortality: No animal died in any group.Clinical signs: Salivation was observed in six males (main group) receiving 300 mg/kg bw/day on week 4 of the dosing period. This effect was considered to be due to irritating property of the test substance.
DETAILED CLINICAL OBSERVATIONS, MANIPULATIVE TEST, GRIP STRENGTH TEST AND LOCOMOTOR ACTIVITY MEASUREMENT:
Home cage observation: No effects
In-the-hand observation: No effects
Open field observation: No effects
Manipulative test: A significant decrease in landing foot splay was observed in females receiving 300 mg/kg bw/day on Day 4 of the lactation period, but it was considered to be spontaneous.
Measurement of grip strength: No effects
Measurement of Motor Activity: No effects
BODY WEIGHT: No effects
FOOD CONSUMPTION: No effects
URINALYSIS: No effects
HAEMATOLOGY: Significant decreases in hemoglobin and hematocrit levels were observed in females receiving 300 mg/kg bw/day at the end of the dosing period. Significant increases in hemoglobin and hematocrit levels and a significant decrease in reticulocyte counts were observed in females receiving 300 mg/kg bw/day at the end of the recovery period.
CLINICAL CHEMISTRY: A significant increase in ALT and a decrease in total protein level were observed in males receiving 300 mg/kg bw/day at the end of the dosing period.
ORGAN WEIGHTS: There were no changes related to the test substance in any group during the dosing and recovery periods.
GROSS PATHOLOGY:
At the End of the Administration Period
Cecum: Dilatation was observed in eight males and two females receiving 300 mg/kg bw/day.
Stomach: Thickening in limiting ridge was observed in four males and two females receiving 300 mg/kg bw/day.
At the End of the Recovery Period
There were no lesions related to the test substance in any group.
HISTOPATHOLOGY: NON-NEOPLASTIC:
At the End of the Administration Period
Both sexes had minimal hypertrophy of the duodenal mucosal epithelia at 300 mg/kg bw/day
Intestine: Diverticulum in ileum was observed in one control male.
Kidney: Cyst was observed in one control male.
Spleen: Focus in raised was observed in one control male.
At the End of the Recovery Period
There were no lesions related to the test substance in any group.

Based on the changes in the blood and gastrointestinal organs, the NOAEL of repeated dose toxicity was determined to be 60 mg/kg bw/day in male and female rats.

UUID

IUC5-4d2b2168-6941-4ce9-b59d-04e3420efc36

Dossier UUID

0

Author

dra / National Institute of Health Sciences / Tokyo / Japan

Date

2017-01-18 16:47:14 JST

Remarks

Study result type

experimental result

Reliability

1 (reliable without restriction)

Rationale for reliability incl. deficiencies

OECD Test Guideline study under GLP condition