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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
screening for reproductive / developmental 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

Data source

Reference
Reference Type:
other: Autharized database
Title:
Reproduction Toxicity Screening Preliminary Test of 2-Amino-5-Methylbenzenesulfonic Acid by Oral Administration in Rats
Author:
J-CHECK
Year:
2016
Bibliographic source:
Ministry of Health, Labour and Welfare", "Ministry of the Environment" and "National Institute of Technology and Evaluation, J-CHECK - (2016)

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 2-Amino-5-Methylbenzenesulfonic Acid in Rats
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report):2- Amino-5-Methylbenzenesulfonic Acid- Molecular formula (if other than submission substance):C7H9NO3S- Molecular weight (if other than submission substance):187.2181g/mole- Substance type:Organic - Physical state:fine yellowish white powder- Impurities (identity and concentrations):0.7 %

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: Charles River Co., Ltd. (Kanagawa)- Age at study initiation: 10 weeks - Weight at study initiation: 375 to 414 g in males and 239 to 266 g in females - Fasting period before study: No data available - Housing: Individual 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 until nursing 4th. - Diet (e.g. ad libitum): NMF solid feed (radiation sterilized feed) manufactured by Oriental Yeast Co., Ltd., and was taken free during the breeding period. - Water (e.g. ad libitum): Tap water, ad libitum- Acclimation period: 5 daysENVIRONMENTAL CONDITIONS- Temperature (°C): 24 ± 2 ° C- Humidity (%): 55 ± 10% - Air changes (per hr): Ventilation frequency of 15 times / hour,- Photoperiod (hrs dark / hrs light): 12 hours (7 am lights, 7 pm off)

Administration / exposure

Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
silicone oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Administration solution, which was prepared by suspending the pharmacopoeial sesame oil (Miyazawa Pharmaceutical), was sealed stored under the cold shielding until use. Test substance drug substance and administration solution of the test substance was confirmed to be stable.VEHICLE- Justification for use and choice of vehicle (if other than water):- Concentration in vehicle: 0,100 ,300 and 1000 mg/kg/bw/day - Amount of vehicle (if gavage): 0.5 ml per body weight 100 g. - Lot/batch no. (if required): No data available.- Purity:
Details on mating procedure:
- M/F ratio per cage:1;1- Length of cohabitation:14 days - Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancySperm confirmation in vaginal plaque, and that day was taken as the 0 day of pregnancy.
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 2amino5methylbenzenesulfonic acid was contained in the administration liquid used.
Duration of treatment / exposure:
Male: 48 days Female: 69 days
Frequency of treatment:
Daily (Once)
Details on study schedule:
- Dose selection rationale: A 14 days repeated oral toxicity Study was observed in 1 group 4 males and 4 females, and the 0,100,250,500,1000 and 2000 mg / kg / day dose was administered for 14 day daily by oral gavage. General state, body weight, food consumption, urinalysis, hematology testing, in the blood biochemical examination and organ weight, suggest change the toxicity of the test substance was observed. In the autopsy, the cecum of expansion was observed in male and female in all cases of 2000 mg / kg group. Therefore the final dose selected for this study was 100, 300 and 1000 mg/kg/bw/day.
Doses / concentrations
Remarks:
Doses / Concentrations: 0, 100, 300 and 1000 mg/kg/bw/day Basis:
No. of animals per sex per dose:
Total no of animals 96 0 mg/kg/bw/day- 12 male, 12 female100 mg/kg/bw/day - 12 male, 12 female300 mg/kg/bw/day - 12 male, 12 female1000 mg/kg/bw/day – 12 male, 12 female
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available

Examinations

Parental animals: Observations and examinations:
Survival, clinical sign, Body weight and Food intake was observed.
Estrous cyclicity (parental animals):
Copulation formation, average season cycle, number of days between the estrus period and the next estrus period as the sex cycle days and mating rate were examined. Corpus luteums, number of implantation traces were examined.
Sperm parameters (parental animals):
No data available
Litter observations:
Number of births, sex, body weight of 0 and 4 days were examined.
Postmortem examinations (parental animals):
Organ weight, gross pathology and histopathology were examined.
Postmortem examinations (offspring):
Gross abnormalities of organs and tissues were observed.
Statistics:
Weight, food intake, 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 24)was performed on the survival rate, organ weight and relative weight of the rats.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, the average for one newborn baby during the nursing period was taken as one sample. The level of significance was set at two levels of *: P <0.05 and **: P <0.01.
Reproductive indices:
Pregnancy period, Implantation rate, delivery rate, live birth rate, mating rate were examined.
Offspring viability indices:
Viability on day 4 were examined.

Results and discussion

Results: P0 (first parental animals)

General toxicity (P0)

Clinical signs:
no effects observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
effects observed, treatment-related

Reproductive function / performance (P0)

Reproductive function: estrous cycle:
no effects observed
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed

Details on results (P0)

Mortality: No effect on survival of treated male and female rats were observed as compared to control.Clinical signs: No clinical signs were observed in treated male and female rats due to treatment as compared to control.In amle rats, ocular secretion in 1 rat at 100 mg / kg, hair loss in 1 rat at 300 mg / kg and Crust and hair loss were observed in 1 in case at 1000 mg / kg. In female rats, hair removal was observed in 100 mg / kg group through pregnancy and nursing periods, and in the control group, all the dead animals were found in 2 cases. Body weight: No significant effect on body weight and body weight gain were observed in treated male rats as compared to control.In female rats, at 100 and 1000 mg / kg, statistically significant decrease in body weight only at 4th day of nursing was observed as compared to the control. However, since there was no obvious difference on the other measurement days 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: In male rats, statistically significant increase in food consumption from 8 to 15 days cumulative food intake from 1 to 15 days were observed at 1000 mg / kg bw.In female rats, no significant effect was observed as compared to control. Reproductive function: estrous cycle: No effect on estrous cycle were observed in treated rats as compared to contorl. Reproductive performance: No effet on number of corpusluteums, number of implantation traces, number of births and number of stillborn were observed in treated rats as compared to control. Gestation index, Implantation rate, delivery rate, live birth rate and mating rate of treated rats as compared to control. Organ weights: When treated with 300 and 1000 mg/kg bw, statistically significant decrease in absolute weights of epididymis were observed in male rats as compared to control. No effect on relative weight of epididymis were observed as compared to contorl. No significant changes were observed in testicular weigth of treated male rats as compared to control.Gross pathology: In male rats, When treated with 300 mg/kg bw, black lesions on lung in 1 animal, red spots of the liver, testis, epididymis and thinning of the coat in 1 case of the same individual gropue were observed. When treated with 1000 mg/kg bw, thinning of the coat 1 case of the same individual gropue were observed.In female rats, black spots in lung stomach ulcers and white spots of the adrenal glands in 1 case each at 1000 mg/kg bw and ovary cyst and coat thinning were observed in 1 case each at 100 mg/kg bw. No findings that could be attributed to administration of the test substance were observed in any of the animals. Histopathology: In male rats, one case skin erosion and squamous epithelium hyperplasia and cell infiltration of epididymis at 1000 mg / kg, seminiferous tubular, testicular and epididymal atrophy of testes, stromal cell proliferation, spermatozoa of epididymis, lung inflammation and liver necrosis at 300 mg / kg were observed. However, since it is low frequency expression, it was not considered to be influence of the test substance. In female rats, 1 case of the lung inflammation, 1 case of the stomach ulcer, 1 part of the adrenal cortex and atrophyof the thymus at 1000 mg/kg bw, 1 case of atrophy of the thymus at 300 mg/kg bw and Young yolk sac cysts in one case, inflammatory infiltration of the skin and squamous epithelium proliferation were observed in one at 100 mg/kg bw, spontaneously delivered, so it was considered that there was no effect on the ovaries of the test substance. Atrophy of the thymus and purulent inflammation of the uterus were observed in one case in all the dead animals. No abnormal findings were observed in the other case.

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:
other: No effect on Survival, clinical sign, Body weight and Food intake, reproductive performance, organ weight, gross pathology and histopathology
Remarks on result:
other: not specified

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
not specified
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
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P1)

Reproductive function: estrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
not specified

Effect levels (P1)

Dose descriptor:
other: not specified
Based on:
not specified
Sex:
not specified
Remarks on result:
other: not specified

Target system / organ toxicity (P1)

Critical effects observed:
not specified
System:
other: not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
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
Histopathological findings:
no effects observed
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Details on results (F1)

Mortality: No effect on number of live pups were observed as compared to contorl. Body weight: No effect on body weight of pups was observed as compared to control. Gross pathology: No gross pathological changes were observed in pups as compared to control.Histopathology: Renal pelvic enlargement in 1 case, renal pelvic enlargement in the kidney It was recognized in 1 and 3 cases in control and 300 mg / kg respectively. In both cases, the expression was expressed in a few cases, which was not related to the administration of the test substance.

Effect levels (F1)

Dose descriptor:
other: not specified
Generation:
other: not specified
Based on:
not specified
Sex:
not specified
Remarks on result:
other: not specified

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F2 generation

General toxicity (F2)

Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
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:
not specified
Histopathological findings:
not specified
Other effects:
not specified

Developmental neurotoxicity (F2)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F2)

Developmental immunotoxicity:
not specified

Effect levels (F2)

Dose descriptor:
other: not specified
Generation:
other: not specified
Based on:
not specified
Sex:
not specified
Remarks on result:
other: not specified

Target system / organ toxicity (F2)

Critical effects observed:
not specified
System:
other: not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables

Absolute and relative organ weight of rats treated orally with 2-Amino-5-methylbenzenesulfonic acid in the preliminary reproduction toxicity screening test.

Dose levelmg/kg/day

0

100

300

1000

Male

No. of animals examined

12

12

12

12

Body weight(g)

542±33

554±45

534±36

546±3

Absolute organ weight

 

 

 

 

Testes (g)

3.62±0.31

3.65±0.44

3.28±0.59

3.70±0.23

Epididymides(mg)

1313±98

1295±138

1168±121*

1316±115

Relative organ weight

 

 

 

 

Testes (g %)

0.671±0.074

0.664±0.094

0.616±0.119

0.680±0.666

Epididymides

 (mg %)

243.378±27.682

235.898±34.456

219.293±27.989

242.189±27.983

Values are expressed as Mean±SD

Significant difference from control group*P<0.05

Summary of reproductive performance in rats treated orally with2-Amino-5-methylbenzenesulfonic acid in the preliminary reproduction toxicity screening test

Dose levelmg/kg/day

0

100

300

1000

No. of pairs mated

12

12

12

12

No. of pairs copulated

12

12

12

12

No. of pregnant female

11

10

12

12

Copulation index%a

100.0

91.7

100.0

100.0

Fertility index %b

91.7

90.9

100.0

100.0

Estrus cycle( days ,Mean±SD)

4.5±0.7

4.2±0.5

4.2±0.4

4.5±0.5

a-No. of animals with successful copulation/No. of animals mated) X100

b-No. Of pregnant animals/ No. of animals with successful copulation X 100

Values in parentheses are expressed no. of animals observed

Finding of delivery in dams treated orally with -Amino-5-methylbenzenesulfonic acid and observation on their pups(F1) in the preliminary reproduction toxicity screening test

Dose level mg /kg/day

0

100

300

1000

No. of dams observed

11

10

12

12

No. of dams delivered live pups

11

10

12

12

Duration of gestation (Mean±SD)

22.7±0.6

22.4±0.5

22.3±0.5

22.7±0.4

No. of Corpora lutea(Mean±SD)

216

170

218

222

No. of total implants(Mean±SD)

188

161

186

175

No of total implants(Mean±SD)

172

150

178

160

No. of total pups born(Mean±SD)

168

150

178

160

Male

81

69

91

79

Female

87

81

87

81

Sex ratio (male and female) (Mean±SD)

1.00

0.93

1.13

1.21s

No of total live pups on days 4(Mean±SD)

 

 

 

 

Male

66

66

85

78

Female

66

77

80

79

No. of total dead pups born(Mean±SD)

4

0

0

0

Still birth

0

0

0

0

Cannnitelism

4

0

0

0

Gestation index(Mean±SD)a

100

100

100

100

Implantation index(Mean±SD)b

89.5±12.3

94.7±2.0

86.6±15.8

80.2±18.2

Delivery index(Mean±SD)c

91.6±5.6

93.5±7.7

96.2±4.8

90.9±8.6

Live birth index(Mean±SD)d

97.4±8.6

100±0

100±0

100±0

Viability index on day 4(Mean±SD)e

Male

77.8±39.1

96.08.4

93.1±9.7

99.1±3.2

Female

77±39.1

95.7±5.6

94.4±15.8

98±4.8

a-(No of females with live pups/ No. of pregnant females) x100

b-( No of implants/No. of corpora lutea)x100

c-(( No .of pups born/No. of implants)X100

d.(No. of live pups born / No. of pups born)x100

e.-No. of live pups on day4 after birth/No. of live pups born)X100

(Includes live pups died before observation)

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 1000 mg/kg bw for P and F1 generation when Crj: CD (SD) male and female rats were treated with 2- Amino-5-Methylbenzenesulfonic Acid orally by gavage in Sesame oil for 69 days.
Executive summary:

In a reproductive toxicity study,Crj: CD (SD) male and female rats were treated with2- Amino-5-Methylbenzenesulfonic Acid in the concentration of0, 100, 300 and 1000 mg/kg/bw/day orally by gavage in Sesame oil. No effect on survival of treated male and female rats were observed. In amle rats, ocular secretion in 1 rat at 100 mg /kg, hair loss in 1 rat at 300 mg /kg and Crust and hair loss were observed in 1 in case at 1000 mg/kg. In female rats, hair removal was observed in 100 mg/kg group through pregnancy and nursing periods, and in the control group, all the dead animals were found in 2 cases. No clinical signs were observed in treated male and female rats due to treatment as compared to control.In female rats, at 100 and 1000 mg / kg, statistically significant decrease in body weight only at 4th day of nursing was observed as compared to the control. However, since there was no obvious difference on the other measurement days 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, statistically significant increase in food consumption from 8 to 15 days cumulative food intake from 1 to 15 days were observed at 1000 mg / kg bwandin female rats, no significant effect was observed as compared to control. Similarly, no reproductive toxic effect were observed onestrous cycle, corpus luteums, number of implantation traces, number of births and number of stillborn, gestation index, implantation rate, delivery rate, livebirth rate and mating rate of treated rats as compared to control. In addition,statistically significant decrease in absolute weights of epididymiswere observedin male rats but no effect on relative weight of epididymis were observed at 300 and 1000 mg/kg bw as compared to contorl.No significant changes were observed intesticular weigth of treated male rats as compared to control. In male rats, black lesions on lung in 1 animal, red spots of the liver, testis, epididymis and thinning of the coat in 1 case of the same individual gropue were observed at 300 mg/kg bw and thinning of the coat 1 case of the same individual gropue were observed at 1000 mg/kg bw. In female rats, black spots in lungstomach ulcers and white spots of the adrenal glands in 1 case each at 1000 mg/kg bw and ovary cyst and coat thinning were observed in 1 case each at 100 mg/kg bw. No findings that could be attributed to administration of the test substance were observed in any of the animals. In male rats, one case skin erosion and squamous epithelium hyperplasia and cell infiltration of epididymis at 1000 mg / kg, seminiferous tubular, testicular and epididymal atrophy of testes, stromal cell proliferation, spermatozoa of epididymis, lung inflammation and liver necrosis at 300 mg / kg were observed. However, since it is low frequency expression, it was not considered to be influence of the test substance. In female rats, 1 case of the lung inflammation, 1 case of the stomach ulcer, 1 part of the adrenal cortex and atrophy of the thymus at 1000 mg/kg bw, 1 case of atrophy of the thymus at 300 mg/kg bw and Young yolk sac cysts in one case,inflammatory infiltration of the skin and squamous epithelium proliferation were observed in one at 100 mg/kg bw, spontaneously delivered, so it was considered that there was no effect on the ovaries of the test substance. Atrophy of the thymus and purulent inflammation of the uterus were observed in one case in all the dead animals. No abnormal findings were observed in the other case. Therefore, NOAEL was considered to be 1000 mg/kg bw for P and F1 generation when Crj: CD (SD) male and female rats were treated with2- Amino-5-Methylbenzenesulfonic Acid orally by gavage inSesame oil for 28 days.