Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from J-check

Data source

Reference
Reference Type:
review article or handbook
Title:
Simple reproductive toxicity test by oral administration of 2-amino-5- methylbenzenesulfonic acid in rats
Author:
J-check
Year:
2010
Bibliographic source:
Ministry of Health, Labour and Welfare", "Ministry of the Environment" and "National Institute of Technology and Evaluation, J-check- 2010

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
Reproduction Toxicity Screening Test of test chemical by Oral Administration in Rats
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report):2-Amino-5-methylbenzenesulfonic acid
- Molecular formula :C7H9NO3S
- Molecular weight :187.2181 g/mole
- Substance type:Organic

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Charles River KK (Kanagawa)
- Age at study initiation: (P) x wks; (F1) x wks: 10 weeks
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: 375 to 414 g for males and 239 to 266 g for females.
- Fasting period before study:
- Housing: One animal was housed and housed in an aluminum front and floor stainless steel mesh breeding cage in a breeding room. Maternal animals after gestation day 18 were kept on an aluminum front and floor stainless steel mesh breeding cage with nursery tray and nest material (manufactured by Care FRESHTM, Absorption corporation) until nursing 4th.
- Diet (e.g. ad libitum): NMF solid feed (radiation sterilized feed) and was taken free during the breeding period
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 2 ° C
- Humidity (%): 55 ± 10%
- Air changes (per hr): 15 times / hour
- Photoperiod (hrs dark / hrs light): illuminance of 150 to 300 lux, illumination time of 12 hours (7 am lights, 7 pm off)

Administration / exposure

Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: sesame oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The test substance was suspended in sesame oil (manufactured by Nacalai Tesque, Inc.), and each group of administration liquids was prepared so as to have concentrations of 20, 60 and 200 mg / mL. After preparation, it was kept under light-shielded and refrigerated conditions until use. It was confirmed that the test substance in the administration solution was stable for at least 7 days under light shielded and refrigerated conditions at concentrations of 6 and 200 mg / mL.

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): sesame oil
- Concentration in vehicle: 0 (vehicle), 100, 300, 1000 mg/kg/day
- Amount of vehicle (if gavage): 0.5 mL per 100 g body weight
- Lot/batch no. (if required):
- Purity:
Details on mating procedure:
- M/F ratio per cage: 1: 1.
- Length of cohabitation: Every night for a maximum of 14 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Sperm in vaginal plaque, and that day was taken as the 0th day of pregnancy.
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility. Not specified
- Further matings after two unsuccessful attempts: [no / yes (explain)] Not specified
- After successful mating each pregnant female was caged (how): Not specified
- Any other deviations from standard protocol: Not specified
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For the analysis of the concentration and homogeneity of the administration solution, samples were randomly extracted from batches of each group prepared at the start of preparation. As a result, the error with respect to the display density was in the range of -12.5 to -0.4% and within the reference range (within ± 15%). Therefore, it was confirmed that a prescribed amount of 2-amino-5-methylbenzenesulfonic acid was contained in the administration solution used.
Duration of treatment / exposure:
For male- 48 days
For female – approx 68 days
Frequency of treatment:
Daily
Details on study schedule:
Not specified
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day
Dose / conc.:
100 mg/kg bw/day
Dose / conc.:
300 mg/kg bw/day
Dose / conc.:
1 000 mg/kg bw/day
No. of animals per sex per dose:
Total: 96
0 mg/kg/day: 12 male, 12 female
100 mg/kg/day: 12 male, 12 female
300 mg/kg/day: 12 male, 12 female
1000 mg/kg/day: 12 male, 12 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Doses were based on 28-day repeated dose toxicity study at a concentration of 0, 30, 100, 300 and 1000 mg / kg, increased the urine specific gravity, urine pH, Reduction or decrease in white blood cell count and total cholesterol was observed in females with an increase in GPT and a decrease in blood glucose, and autopsy revealed cecal expansion in both males and females. Based on these results, a preliminary test carried out at doses of 0, 100, 300 and 1000 mg / kg. Therefore, as in the preliminary test, high dose of 1000 mg / kg was also set, divided by the common ratio of about 3, and 300 and 100 mg / kg were set.
- Rationale for animal assignment (if not random): The animals were stratified based on the body weight at the start of administration, and 12 animals per group were sorted by a random sampling method.
- Other:

Examinations

Parental animals: Observations and examinations:
Survival, general condition, body weights, food consumption were examined.
Oestrous cyclicity (parental animals):
Estrous cyclicity were examined.

Number of corpus luteum and the number of implantation were examined.
Sperm parameters (parental animals):
Not specified
Litter observations:
Number of births, clinical signs and body weight were examined.
Postmortem examinations (parental animals):
Organ weight, Gross pathology and histopathology were examined.
Postmortem examinations (offspring):
Gross pathology were examined.
Statistics:
Weight, food consumption, number of corpus luteums, number of implantation traces, number of births, number of stillbirths, sex ratio, average sex, pregnancy period, implantation rate, delivery rate, birth rate, abnormal incidence of abnormal outer table, newborn 4th Multiple comparison test 2-4) was performed on the survival rate, organ weight and relative weight of the subjects. For the birth rate, mating rate and conception rate, χ ^ 2 tests 5, 6) were used. The incidence of findings of pathological examination was tested using Fisher's direct probability test method 6). In addition, as for the results on newborns during the nursing period, the average per mother was taken as one sample. The level of significance was set at two levels of *: P <0.05 and **: P <0.01.
Reproductive indices:
Copulation index, fertility index, implantation index, gestation index, delivery index were examined.
Offspring viability indices:
viability index on day 0 and 4 were examined.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
In male rats, 1 ocular secretion in the 100 mg / kg group of male, 1 hair loss in the 300 mg / kg group, 1 in the 1000 mg / kg group Crust and hair loss were observed in one case.
In females, hair removal was observed in the 100 mg / kg group through pregnancy and nursing periods
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No mortality were observed in treated male and femlae rats as compared to control.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
In male, no effect on body weight were observed as compared to control. In female at 100 and 1000 mg / kg, statistically significant decrease in body weight were observed only on the 4th day of nursing compared to the control group. However, since there was no obvious difference on the other measurement day and it was a change only for 1 day, and since there was no obvious change in body weight gain during the nursing period, the influence of administration of the test substance It was not thought to be an accidental change.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
In male rats at 1000 mg/kg bw, statistically significant increase in daily food intake for the 8 to 15 days and the cumulative food consumption from 1 to 15 days were observed as compared to control.
In female rats, no effect on food consumption was observed as compared to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
When treated with 1000 mg/kg bw, cell infiltration of the epididymis, case in skin erosion and squamous cell epithelial hyperplasia , 1 atrophy of the thymus, 1 lung inflammation, 1 stomach ulcer and 1 part of the adrenal cortex one hypertrophy were observed in male rats.
When treated with 300 mg/kg bw, seminiferous tubular atrophy of the testes, pulmonary inflammation, 1 case in the liver necrosis and 1 atrophy of the thymus were observed in male rats.

When treated with 100 mg/kg bw, 1 Young yolk sac cysts in females who spontaneously delivered, so it was considered that there was no effect on the ovaries of the test substances. 1 Skin inflammatory infiltration and squamous epithelial hyperplasia were observed.

In males and females who did not mate, seminiferous tubular atrophy of the testes was observed in males. No abnormal findings were observed in females.

No abnormal findings were observed in males who did not establish pregnancy and infertile females.

Atrophy of the thymus and purulent inflammation of the uterus were observed in one case in all the dead animals. No abnormal finding was observed in the other case.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
No effect on copulation, implantation and sexual cycle of treated female rats were observed as compared to control.
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
No effect on gestation length, number of corpora lutea or implantations, implantation index, gestation index, delivery index, purturition or maternal behavior, numbers of offspring or live offspring, sex ratio and live birth index were observed in treated rats as compared to control.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (oestrous cycle)
reproductive performance
other: No effect

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Description (incidence and severity):
No Clinical sign were observed in neonates as compared to control.
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
No effect on Viability of neonates were observed on day 0 and 4 as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No effect on Body weight of neonates were observed on day 0 and 4 as compared to control.
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
No abnormal findings were observed in females.
At autopsy on 4th day of nursing, in the male, thymus neck remnant was 1 in the control group, 1 in the renal pelvic enlargement in the 300 mg / kg group, in the female in the control group and in the 300 mg / kg group in the renal pelvic enlargement in the kidney It was recognized in 1 and 3 cases respectively. In both cases, expression was expressed in a few cases, which was not related to administration of the test substance.
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

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:
viability
clinical signs
mortality
body weight and weight gain
gross pathology
other: No effect observed

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 1000 mg/kg/day for P and F1 generation when male and female rats treated with test chemical orally by gavage for approx 68 days.
Executive summary:

In a Preliminary Reproduction Toxicity Screening Test,Crj:CD(SD) male and female rat were treated with test chemical in the concentration of 0 (vehicle), 100, 300, 1000 mg/kg/day orally by gavage in sesame oil for male consecutive 48 days of 14 days before mating and 14 days of mating period and 20 days after the end of the mating period. The female administration period was 14 days before the mating and during the mating period (maximum 14 days) and through 3 days of postpartum feeding (41 to 46 days) throughout the gestation period of the female. The females who did not deliver after copulation establishment were for 41 and 43 days up to the day before dissection on 25th gestation. Females that failed to mate were consecutive 48 days of 20 days after the mating period. No mortality was observed in treated male and female rats as compared to control. In male rats, 1 ocular secretion in the 100 mg / kg group of male, 1 hair loss in the 300 mg / kg group, 1 in the 1000 mg / kg group Crust and hair loss were observed in one case and in females, hair removal was observed in the 100 mg / kg group through pregnancy and nursing periods. In male, no effect on body weight and in female at 100 and 1000 mg / kg, statistically significant decrease in body weight were observed only on the 4th day of nursing compared to the control group. However, since there was no obvious difference on the other measurement day and it was a change only for 1 day, and since there was no obvious change in body weight gain during the nursing period, the influence of administration of the test substance It was not thought to be an accidental change.In male rats at 1000 mg/kg bw, statistically significant increase in daily food intake for the 8 to 15 days and the cumulative food consumption from 1 to 15 days and in female rats, no effect on food consumption was observed as compared to control. Similarly,No reproductive effect on copulation, implantation and sexual cycle, gestation length, number of corpora lutea or implantations, implantation index, gestation index, delivery index, purturition or maternal behavior, numbers of offspring or live offspring, sex ratio and live birth index were observed in treated rats as compared to control. At 300 and 1000 mg/kg bw, statistically significant decrease in absolute epididymis weight were observed but no relative weight change and no effect on testicular weight were observed in treated male rats as compared to control. No gross pathological changes were attributed to the administration of the test substance. At 1000 mg/kg bw, cell infiltration of the epididymis, case in skin erosion and squamous cell epithelial hyperplasia , 1 atrophy of the thymus, 1 lung inflammation, 1 stomach ulcer and 1 part of the adrenal cortex one hypertrophy were observed in male rats. At 300 mg/kg bw, seminiferous tubular atrophy of the testes, pulmonary inflammation, 1 case in the liver necrosis and 1 atrophy of the thymus were observed in male rats. At 100 mg/kg bw, 1 Young yolk sac cysts in females who spontaneously delivered, so it was considered that there was no effect on the ovaries of the test substances. 1 Skin inflammatory infiltration and squamous epithelial hyperplasia were observed. In males and females who did not mate, seminiferous tubular atrophy of the testes was observed in males. No abnormal findings were observed in females. No abnormal findings were observed in males who did not establish pregnancy and infertile females. Atrophy of the thymus and purulent inflammation of the uterus were observed in one case in all the dead animals. No abnormal finding was observed in the other case. In addition,No effect on Viability and body weight of neonates were observed on day 0 and 4 andnoclinical sign were observed inneonates as compared to control.No abnormal findings were observed in females. At autopsy on 4th day of nursing, in the male, thymus neck remnant was 1 in the control group, 1 in the renal pelvic enlargement in the 300 mg / kg group, in the female in the control group and in the 300 mg / kg group in the renal pelvic enlargement in the kidney It was recognized in 1 and 3 cases respectively. In both cases, expression was expressed in a few cases, which was not related to administration of the test substance. Therefore, NOAEL was considered to be 1000 mg/kg/day for P and F1 generation when male and female rats treated with test chemical orally by gavage for approx68 days.