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Repeated dose toxicity: oral

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Administrative data

Endpoint:
repeated dose toxicity: oral, other
Remarks:
Combined repeated dose reproductive-develpmental screening
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:
other: J-check
Title:
Combined repeated dose reproductive-develpmental screening for CAS no 88-44-8
Author:
National Institute of Technology and Evaluation
Year:
2018
Bibliographic source:
Japan Chemicals Authoritative Knowledge Database, 2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Refer below principle
Principles of method if other than guideline:
Combined repeated dose reproductive-develpmental screening was performed to evaluate the toxic nature of 2-amino-5-methyl benzene sulfonic acid in male and female Crj: CD (SD) 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
Specific details on test material used for the study:
- Name of test material:2-Amino-5-methylbenzenesulfonic acid
- Molecular formula :C7H9NO3S
- Molecular weight :187.2181 g/mol
- Substance type: Organic
- Physical state:Yellow powder

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Details on species / strain selection:
No data
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River KK (Kanagawa)
- Age at study initiation: 9 weeks old approx..
- Weight at study initiation: 375 to 414 g in males and 239 to 266 g in females
- Fasting period before study:
- Housing: The animals were housed in aluminum front and floor stainless steel mesh breeding cage in a breeding room.
- Diet (e.g. ad libitum): NMF solid feed (radiation sterilized feed) ad libitum
- 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): 150 to 300 lux, for 12 hours (7 am lights, 7 pm off)

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
No data
Vehicle:
other: sesame oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with sesame oil at dose levels of 0, 100 ,300 and 1000 mg/kg/bw/day

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

VEHICLE
- Justification for use and choice of vehicle (if other than water): Sesame oil
- Concentration in vehicle: 0, 100 ,300 and 1000 mg/kg/bw/day
- Amount of vehicle (if gavage): 0.5 mL/100 g bw
- Lot/batch no. (if required): No data available.
- Purity: No data
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
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.

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:
Males, 48 days
Females, from 14 days before mating to day 3 of lactation
Frequency of treatment:
Daily (Once), 7 day/week
Doses / concentrations
Remarks:
0, 100 ,300 and 1000 mg/kg/bw/day
No. of animals per sex per dose:
0 mg/kg/bw/day- 12 male and 12 female
100 mg/kg/bw/day - 12 male and 12 female
300 mg/kg/bw/day - 12 male and 12 female
1000 mg/kg/bw/day - 12 male and 12 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: On the basis of preliminary 2 week reproductive study conducted, the doses for the main study was set as 0, 3, 300 or 1000 mg/Kg/day
- 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.
- Rationale for selecting satellite groups: No data
- Post-exposure recovery period in satellite groups: No data
- Section schedule rationale (if not random): No data
Positive control:
No data

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked in table [No.?] were included. General condition

DETAILED CLINICAL OBSERVATIONS: Yes / No / No data
- Time schedule:

BODY WEIGHT: Yes
- Time schedule for examinations: In male, the body weight gain from day 1 to day 43 of administration was calculated by measurement 1 (administration start date), 8, 15, 22, 29, 36, 43 and 49 days (autopsy day). For females, body weight gain was measured from 1st to 15th days of dosing as measured on Dosage 1 (administration start date), 8 and 15 days.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes, For males, the bait weight was measured on the administration 1 (administration start date), 8, 15, 22, 29, 36, 43 and 48 days (the day before necropsy), and the food intake amount from the measurement date to the next measurement day To calculate the average daily feed amount and calculate cumulative food intake from 1 to 15 days of administration and from 22 to 48 days of administration. In females, on the 8th and 15th dosing 1
(administration initiation day), females who did not mate were measured for bait weight at the subsequent doses
of 29, 36, 43 and 48 (the day before autopsy) The average food intake was calculated and the cumulative food intake from 1 to 15 days of administration was calculated.
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: No data
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

CLINICAL CHEMISTRY: Y No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Males: On the day following 48 days of administration, blood was euthanized by exsanguination under ether anesthesia. Organs / tissues were observed with the naked eyes, and the weight of testis and epididymis was measured to calculate organ weight / body weight ratio (relative weight). In addition, lungs, liver and skin were fixed with 10% neutral buffered formalin solution as organs / tissues in which changes were observed in seminal vesicles, prostate and macroscopic findings of all animals. The testis and epididymis were fixed with Bouin's solution.

Females: On nursing 4th day, exsanguination was euthanized by ether anesthesia. After macroscopic observation of organs and tissues, lung, skin, thymus, stomach and adrenal glands were fixed with 10% neutral buffered formalin solution as organs / tissues whose changes were observed by ovary, uterus, vagina and macroscopic findings. In addition, at the time of necropsy, the number of corpus luteum and the number of implantation traces were examined.

HISTOPATHOLOGY: Yes, the animals were subjected to histopathological examination.
Other examinations:
No data
Statistics:
For weight and food consumption 4th Multiple comparison test was performed on the survival rate, organ weight and relative weight of the mice.

Results and discussion

Results of examinations

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No change due to administration of the test substance was observed, 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.
Mortality:
no mortality observed
Description (incidence):
No mortality was reported during the study period
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
In males, no difference was observed between the control group and each test substance-administered group throughout the administration period.

In females, at 100 and 1000 mg / kg group, the statistically significant low values 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 males, the mean daily food intake for the 8 to 15 days in the 1000 mg / kg group showed a statistically significant high value, and the cumulative food consumption from 1 to 15 days of administration showed a high value.

In females, there was no difference between the control group and each test substance-administered group over the administration period.
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:
no effects observed
Description (incidence and severity):
In the 300 mg / kg group, the actual weight of the epididymis showed a statistically significant lower value than the control group, but there was no difference in the relative weight, and a similar change was observed in the 1000 mg / kg group. There was no difference between the control group and each test substance administered group for testicular weight.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No findings that could be attributed to the administration of the test substance were observed in any of the animals. In males, black lesions of the lungs were observed in the control group and 1 group in the 300 mg / kg group in the male, 1 red subject in the liver and white spots in the 300 mg / kg group, 1 in the same individual, testis and epididymal atrophy was observed in one group of the same individual in the 300 mg / kg group, and one in each group in the 300 and 1000 mg / kg group of thinning of the coat.

No abnormal findings were observed in one female in the 100 mg / kg group that did not mate.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Tissue findings showed seminiferous tubular atrophy in male. However, although it is suggested that it may be a factor of mating failure, it was judged that there is no relation with the administration of the test substance from the frequency of occurrence.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Effect levels

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: No significant alterations were noted at the mentioned dose level

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table: Absolute and relative organ weight

Dose level

0

100

300

1000

Males

12

12

12

12

No. of animals examined

542±33

554±45

534±36

546±3

Body weight (g)

3.62±0.31

3.65±0.44

3.28±0.59

3.70±0.23

Absolute organ weight

1313±98

1295±138

1168±121*

1316±115

Testes (g)

0.671±0.074

0.664±0.094

0.616±0.119

0.680±0.066

Epididymides (mg)

243.378±27.682

235.3898±34.456

219.293±27.989

242.189±27.983

Relative organ weight

 

 

 

 

Testes (g)

0.670± 0.074

0.664± 0.094

0.616± 0.119

0.680± 0.066

Epididymides (mg)

243.378± 27.682

235.898± 34.456

219.293± 27.989

242.189± 27.983

* Significant difference from control group P<0.05

Table: Histological findings

Dose level (mg/Kg)

Male animals

Female animals

0

100

300

1000

0

100

300

1000

No. of animals necropsied

11

10

12

12

9

10

12

12

Thymus atrophy

-

-

-

-

-

-

1 (1)

2 (2)

Lung inflammation

1 (1)

-

1 (1)

-

-

-

-

1 (1)

Stomach ulcer

-

-

-

-

-

-

-

1 (1)

Liver necrosis

-

-

1 (1)

-

-

-

-

-

Testis atrophy

0

-

1 (1)

0

-

-

-

-

Interstitial cell hyperplasia testis

0

-

0 (1)

 

-

-

-

-

Ovary cyst, brusa

-

-

-

-

0

1 (1)

-

0

Adrenal gland hypertrophy

-

-

-

-

-

-

-

1 (1)

Skin erosion

-

-

0 (1)

0 (1)

-

0 (1)

-

-

Skin inflammatory infiltration

-

-

0 (1)

1 (1)

-

1 (1)

-

-

Squamous hyperplasia

-

-

0 (1)

1 (1)

-

1 (1)

-

-

 

Applicant's summary and conclusion

Conclusions:
The No observed adverse effect level (NOAEL) was considered to be 1000 mg/kg/day for 2-amino-5-methyl benzene sulfonic acid in male and female Crj: CD (SD) rats during subchronic repeated dose toxicity study.
Executive summary:

Repeated dose toxicity study was performed to evaluate the toxic nature of 2-amino-5-methyl benzene sulfonic acid in male and femaleCrj: CD (SD) rats. The test chemical was dissolved in sesame oil and used at dose levels of 0, 30, 300 or 1000 mg/Kg/day. The treated animals were observed for clinical signs, mortality, changes in body weight and food consumption, and the animals were subjected to gross and histopathology. In both males and females, there was no effect of administration of test substance on change in general condition and body weight.In males, the mean daily food intake for the 8 to 15 days in the 1000 mg / kg group showed a statistically significant high value, and the cumulative food consumption from 1 to 15 days of administration showed a high value. In females, there was no difference between the control group and each test substance-administered group over the administration period. In males, no difference was observed between the control group and each test substance-administered group throughout the administration period. In females, at 100 and 1000 mg / kg group, the statistically significant low values 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 the 300 mg / kg group, the actual weight of the epididymis showed a statistically significant lower value than the control group, but there was no difference in the relative weight, and a similar change was observed in the 1000 mg / kg group. There was no difference between the control group and each test substance administered group for testicular weight. No findings that could be attributed to the administration of the test substance were observed in any of the animals. In males, black lesions of the lungs were observed in the control group and 1 group in the 300 mg / kg group in the male, 1 red subject in the liver and white spots in the 300 mg / kg group, 1 in the same individual, testis and epididymal atrophy was observed in one group of the same individual in the 300 mg / kg group, and one in each group in the 300 and 1000 mg / kg group of thinning of the coat. No abnormal findings were observed in one female in the 100 mg / kg group that did not mate. Tissue findings showed seminiferous tubular atrophy in male. However, although it is suggested that it may be a factor of mating failure, it was judged that there is no relation with the administration of the test substance from the frequency of occurrence. Based on the observations made, the No observed adverse effect level (NOAEL) was considered to be 1000mg/kg/day for 2-amino-5-methyl benzene sulfonic acid in male and female Crj: CD (SD) rats during subchronic repeated dose toxicity study.