1,2-benzisothiazol-3(2H)-one 1,1-dioxide

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

two-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer-reviewed journal

Qualifier:

according to guideline

Guideline:

other: refer below principle

Principles of method if other than guideline:

Two-Generation reproduction toxicity study of Saccharin orally in Charles Rivers (CD)-derived Sprague-Dawley Rats

GLP compliance:

not specified

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles Rivers, Margate, Kent, United Kingdom.
- Age at study initiation: (P) x wks: No data available
(F1) x wks: 22 days old
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: No data available
- Fasting period before study: No data available
- Housing: Animals were housed in groups of three to five per cage for a period of 4 weeks prior to mating, after mating female were housed singly in wire cages for the first 19 days of gestation and then transferred to Perspex boxes with wood shavings. Offspring were housed separately.
- Diet (e.g. ad libitum): Normal diet, ad libitum
- Water (e.g. ad libitum): No data available
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: Normal diet

Details on mating procedure:

Details on study schedule:The offspring were sexed when 22 days old, and the males and females from each litter were housed in separate cages and fed the appropriate diet until killed.
- M/F ratio per cage:1: 1
- After successful mating each pregnant female was caged (how):Singly

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
0 or 5% (2500 mg/kg/day)
Basis:
no data

No. of animals per sex per dose:

No data available

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Parental animals: Observations and examinations:

No data available

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

No data available

Postmortem examinations (parental animals):

Tissue concentrations of saccharin were examined in liver, kidnye, bladder wall, plaema and amnoitic fluid.

Postmortem examinations (offspring):

Tissue concentrations of saccharin were examined in liver, kidney, bladder wall, plaema and amnoitic fluid.

Statistics:

Determined by unpaired Student’s t test

Reproductive indices:

No data available

Offspring viability indices:

No data available

Clinical signs:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

not specified

OTHER FINDINGS (PARENTAL ANIMALS):Tissue Saccharin Concentrations were decrased in female and no no evidence of excessive accumulation in the bladder wall or other tissues of male rats during in utero exposure or during lactationas were observed compared to maternal level.

Dose descriptor:

NOAEL

Effect level:

2 500 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No Effect on organ tissue concentration

OTHER FINDINGS (OFFSPRING):Fetal liver, kidney and bladder wall tissue Saccharin Concentrations were decrased as compared to maternal level

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

2 500 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No Effect on organ tissue concentration

Reproductive effects observed:

not specified

Conclusions:

NOAEL was considered to be 5% (2500 mg/kg/day) in the F0 and F1 generation when male and female Sprague-Dawley rats were treated with saccharin.

Executive summary:

In a two-generation repeated dose toxicity study, male and female Sprague-Dawley rats were treated orally in diet with saccharin in concentration of 0 or 5% (2500 mg/kg/day). In the F0 generation, tissue saccharin concentrations were decreased in female rats and no evidence of excessive accumulation were shown in the bladder wall or other tissues of female rats during in utero exposure or during lactation. The results showed decreased tissue saccharin concentrations in fetal liver, kidney and bladder wall when compared to maternal level.

Therefore, NOAEL was considered to be 2500 mg/kg/day (5%) in the F0 and F1 generation when male and female Sprague-Dawley rats were treated with saccharin

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

2 500 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

Data is from peer-reviewed journal obtained from two generation experimental study

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Short description of key information:
From the data available for experimental studies that have been reported in peer reviewed journals, it is possible to use a weight of evidence appraoch and conclude that in most of the experimental studies no adverse effects on the reproductive organs due to administration of saccharin have been observed as compared to control. Thus, it can be cncluded that saccarin is not expected to cause reproductive toxicity effects at the dose levels reported in the various studies.

Justification for selection of Effect on fertility via oral route:
In a Two-Generation repeapted dose toxicity study, Charles Rivers (CD)-derived Sprague-Dawley  male and female rats treated with Saccharin in concentration of 0 and 2500 mg/kg/day (5%) orally in diet. In F0 generation,Tissue Saccharin Concentrations were decreased in female and no evidence of excessive accumulation in the bladder wall or other tissues of male rats during in utero exposure or during lactationas were observed compared to maternal level and steady-state concentrations of saccharin in the liver and kidneys of fetuses from mothers were observed.

Effects on developmental toxicity

Description of key information

From the various data available and adopting the weight of evidence approach, it has been concluded that 1,2-benzisothiazol-3(2H)-one 1,1-dioxide is not likely to have developmental/teratogenic effects.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviewed journal

Qualifier:

according to guideline

Guideline:

other:

Principles of method if other than guideline:

Teratogenic effects of Saccharin in mouse

GLP compliance:

not specified

Limit test:

no

Species:

mouse

Strain:

other: ICR albino

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:(Camm Research Institute, Wayne, NJ)

Route of administration:

oral: drinking water

Type of inhalation exposure (if applicable):

not specified

Vehicle:

water

Details on exposure:

Details on exposure
PREPARATION OF DOSING SOLUTIONS: No data available

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Water
- Concentration in vehicle: 0, 5, 10 or 20% solution
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on exposure
PREPARATION OF DOSING SOLUTIONS: No data available

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Water
- Concentration in vehicle: 0, 5, 10 or 20% solution
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- M/F ratio per cage: No data available
- Length of cohabitation: No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: No data available
- After … days of unsuccessful pairing replacement of first male by another male with proven fertility: No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]: No data available
- After successful mating each pregnant female was caged (how): No data available
- Any other deviations from standard protocol: No data available

Frequency of treatment:

Daily

Duration of test:

18 days, i.e. from day 0 through day 17 of pregnancy

Remarks:

Doses / Concentrations:
0, 5, 10 or 20% (0, 500, 1000, 2000 mg/Kg bw)
Basis:

No. of animals per sex per dose:

Total: 25 females
Control: 10 females
5%: 5 females
10%: 5 females
20%: 5 females

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Maternal examinations:

All dams were killed by cervical dislocation on day 17 and fetuses removed and examined.

Ovaries and uterine content:

The number of resorptions was observed.

Fetal examinations:

Fetuses were removed from maternal uteri and were examined for gross internal and external malformations. Externally, limbs, back, facies, and head were inspected for gross malformations. Internally, the palate, brain, heart, lungs, kidneys, bladder, gut, and gonads were examined for gross anomalies.

Statistics:

No data available

Indices:

No data available

Historical control data:

No data available

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
Maternal uteri examined for fetal resorption showed no significant increase compared with dams who received no saccharin.

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
None of the fetuses from dams treated with saccharin showed significant increases in either external or internal malformations when compared to controls.

Dose descriptor:

NOAEL

Effect level:

2 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

NOAEL was considered to be 2000 mg/kg bw in female ICR albino mice and their offspring when they the pregnant females were exposed to saccharin

Executive summary:

In a teratogenicity study, the toxic effects of saccharin was evaluated in pregnant female ICR albino mice. The dams received saccharin in drinking water at a dosage of 0, 500, 1000 or 2000 mg/kg bw from day 0 to day 17 of gestation. The results showed no significant increase for fetal resorption in the maternal uteri as compared with dams who received no saccharin. None of the fetuses from dams treated with saccharin in drinking water showed significant increases in either external or internal malformations when compared to control. Therefore, NOAEL was considered to be 2000 mg/kg bw in the maternal F0 generation and in the F1 generation after administration of saccharin in drinking water from day 0 to day 17 of gestation.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

2 000 mg/kg bw/day

Study duration:

subacute

Species:

mouse

Quality of whole database:

The data is K2 level as the data has been obtained from the experimental study from the reliable journal ‘Arch. Toxicol.’.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Weight of evidence summary for reproductive and developmental toxicity

In different studies, saccharin has been investigated to a greater or lesser extent for the reproductive and developmental effects. Often are the studies based on in vivo experiments in rodents, i.e. most commonly in mice or rats.

In a two-generation repeated dose toxicity study by Sweatman et al (1982), male and female Sprague-Dawley rats were treated orally in diet with saccharin in concentration of 0 or 5% (2500 mg/kg/day). In the F0 generation, tissue saccharin concentrations were decreased in female rats and no evidence of excessive accumulation were shown in the bladder wall or other tissues of female rats during in utero exposure or during lactation. The results showed decreased tissue saccharin concentrations in fetal liver, kidney and bladder wall when compared to maternal level.  Therefore, NOAEL was considered to be 2500 mg/kg/day (5%) in the F0 and F1 generation when male and female Sprague-Dawley rats were treated with saccharin

In a multi generation toxicity study conducted by Kroes et al (1977), male and female Swiss SPF-derived outbred mice were treated orally in diet with saccharin in the concentration of 0, 0.2% (286 mg/kg/day) or 0.5% (714 mg/kg/day). No effect were observed on survival, clinical sign and body weight in the offspring of F0 and F1a, F1a’, F2a, F3a, F3b, F4a, F5a or F6a generation as compared to control. No effect were observed on number of implantation, living fetuses per implantation and number of resorption per total number of implantation, gross pathology and histopathology in the F0 generation after treatment with saccharin. There are few effects were observed in F3c, F2b, F4b and F5b compared to control. But the significances found for all investigates parameters were not consistent and are considered not to be due to the treatment at 286 mg/kg/day dose group. Therefore, NOAEL was considered to be 714 mg/kg/day for the F0 generation and 286 mg/kg/day for F1a, F1a’, F2a, F3a, F3b, F4a, F5a and F6a generations when male and female Swiss SPF-derived outbred mice treated with saccharin orally for 21 months. Also based on this study the lowest toxic dose was published to be 16800 mg/kg bw/d of saccharin.

Also from the studies conducted in read across substance sodium salt of saccharin the NOAEL values are found to be 200 mg/kg bw/d and LOAEL was found to be 2866 mg/kg bw/ in two different studies on mice.

Also In a teratogenicity study conducted by Dropkin et al (1984), the toxic effects of saccharin was evaluated in pregnant female ICR albino mice. The dams received saccharin in drinking water at a dosage of 0, 500, 1000 or 2000 mg/kg bw from day 0 to day 17 of gestation. The results showed no significant increase for fetal resorption in the maternal uteri as compared with dams who received no saccharin. None of the fetuses from dams treated with saccharin in drinking water showed significant increases in either external or internal malformations when compared to control. Therefore, NOAEL was considered to be 2000 mg/kg bw in the maternal F0 generation and in the F1 generation after administration of saccharin in drinking water from day 0 to day 17 of gestation.

In the same study the dams received a single i.p. injection of saccharin at a dosage of 0, 500, 1000 or 2000 mg/kg on the 10th day of gestation. The results showed no significant increase for fetal resorption in the maternal uteri as compared with dams who received no saccharin. None of the fetuses from dams treated with saccharin by i.p. injection showed significant increases in either external or internal malformations when compared to control. Therefore, NOAEL was considered to be 2000 mg/kg body weight/day in the maternal F0 generation and in the F1 generation after a single i.p. injection of saccharin on gestation day 10.

Also numbers of teratology studies have been performed in experimental animals using high doses of saccharin by Klotzsche C (1969) and negative results have been documented in rabbits.

Thus based on the above studies it can be conclude that substance saccharin does not have reproductive as well as developmental toxicity effects on higher dose levels as well and considered to be safe.

Justification for selection of Effect on developmental toxicity: via oral route:
NOAEL was considered to be 2000 mg/kg bw in female ICR albino mice and their offspring when they the pregnant females were exposed to saccharin

Justification for classification or non-classification

The chemical 1,2-benzisothiazol-3(2H)-one 1,1-dioxide does not exhibit toxicity to the reproductive system within the doses mentioned in the study end points.