Name:

OECD_SIDS / SUBSTANCE : 2,4-Dimethylbenzenesulfonic acid
dimethylbenzenesulfonic acid / 88-61-9 Tue, 29 Nov 2022, 15:15:34+0900 /Legal entity owner: National Institute of Health Sciences
Printing date: 2022-11-29T15:15:35.007+09:00

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2,4-

Table of Contents
0/0 ............................................................................................................................. 1
National Institute of Health Science ..................................................................... 2
2,4-Dimethylbenzenesulfonic acid ......................................................................... 3
1 General information .................................................................................. 3
1.1 Identification ................................................................................. 3
Identification ............................................................................... 3
Identification ...................................................................... 3
7 Toxicological information ......................................................................... 4
7.5 Repeated dose toxicity ................................................................... 4
7.5.1 Repeated dose toxicity: oral ................................................. 4
Repeated dose toxicity: oral.001 ........................................... 4
7.6 Genetic toxicity ............................................................................ 13
7.6.1 Genetic toxicity in vitro ...................................................... 13
Genetic toxicity in vitro.001 ................................................ 13
Genetic toxicity in vitro.002 ................................................ 19
7.8 Toxicity to reproduction ............................................................... 23
7.8.1 Toxicity to reproduction ..................................................... 23
Toxicity to reproduction.001 ............................................... 23
References ................................................................................................................ 34
Reference Substances ...................................................................................... 34
m-xylene-4-sulphonic acid .......................................................................... 34
Test Materials .................................................................................................. 36
2,4-Dimethylbenzenesulfonic acid ............................................................... 36
Literatures ....................................................................................................... 37
Combined repeated dose toxicity study with the reproductive/
developmental toxicity screening test of 2,4-Dimethylbenzenesulfonic acidby oral administration in rats ..................................................................... 37
In Vitro Chromosomal Aberration Test of on 2,4-Dimethylbenzenesulfonicacid Cultured Chinese Hamster Cells. ......................................................... 38
Reverse Mutation Test of 2,4-Dimethylbenzenesulfonic acid on Bacteria. ...... 39
Legal Entities ................................................................................................... 40
National Institute of Health Sciences .......................................................... 40ii

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Dossier header
Dossier submission type
Name
OECD SIDS
Versioncore 7.0
Name (given by user)

Dossier subject
Dossier subject
2,4-Dimethylbenzenesulfonic acid / 2,4-dimethylbenzenesulfonic acid / 88-61-9
Public nameSubmitting legal entity
National Institute of Health Science
Dossier creation date/time
Tue, 29 Nov 2022, 15:15:34+0900
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LEGAL_ENTITY: National Institute of Health
Science
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General information
Legal entity nameNational Institute of Health Science22,4-Dimethylbenzenesulfonic acid
General information
Identification
Identification
SUBSTANCE: 2,4-Dimethylbenzenesulfonic acid

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Substance name
2,4-Dimethylbenzenesulfonic acid
Legal entity
National Institute of Health Sciences / Kawasaki / Japan

Identification of substance
Reference substancem-xylene-4-sulphonic acid / 2,4-dimethylbenzenesulfonic acid / 88-61-9 / 201-843-9
EC number

EC name201-843-9

EC Inventory

CAS number

CAS name

88-61-9
IUPAC name
2,4-dimethylbenzenesulfonic acid

Role in the supply chain
Manufacturer
false
Importer
false
Only representativefalse
Downstream user
false

3

Toxicological information
Repeated dose toxicity
Repeated dose toxicity: oral
ENDPOINT_STUDY_RECORD: Repeated dose toxicity: oral.001

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Administrative data
Endpoint
short-term repeated dose toxicity: oral
Type of information
experimental study
Adequacy of study
key study
Robust study summary
false
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
other: OECD Test Guideline study under GLP condition
Cross-reference
Reason / purpose for cross-reference
reference to same study 7.8.1 Toxicity to reproduction: Toxicity to reproduction. 001
Related information
OECD / Toxicity to reproduction / Toxicity to reproduction.001 / 2,4-Dimethylbenzenesulfonic acid /2,4-dimethylbenzenesulfonic acid / 88-61-9

Data source
Reference
Combined repeated dose toxicity study with the reproductive/developmental toxicity screening test of /Ministry of Health, Labour and Welfare (MHLW), Japan / study report
Data accessdata published4Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction /Developmental Toxicity Screening Test)
Deviations
yes During the power outage (October 2, 2010) due to the inspection of the electrical equipmen
t of the facility, the humidity increased (78%), the lights were turned off, and the ventilation was
stopped.
However, no effect was observed on the test results.
GLP compliance
yes
Limit test
no

Test material
Test material information
2,4-Dimethylbenzenesulfonic acid
Specific details on test material used for the study- Name of test material (as cited in study report): 2,4-Dimethylbenzenesulfonic acid
- Analytical purity: 99.7% (anhydrous equivalent, contained 16.1% water)- Storage condition of test material: Room temperature, shading- Stability under test conditions: The stability of test material was identified by analysis of the remainder.

Test animals
Speciesratcommon rodent species
Strainother: Crl:CD(SD)Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source: Charles River Japan, Inc., Atsugi Breeding Center.
- Age at study initiation: 10 weeks old- Weight at study initiation:
Male: 355-428 g, Female: 220-249 g- Housing:
Mating period: Stainless steel mesh cages for group housing (340W × 294D × 176H mm)
After 18 days of pregnancy: Stainless steel cage for delivery (340W × 294D × 176H mm) and bedding.
Other breeding period: 2-piece stainless steel mesh cage (170W × 294D × 176H mm/animal)- Diet: Solid feed (CRF-1: Oriental Yeast Co., ltd.) was given ad libitum.
5

- Water: Tap water was given ad libitum.
- Acclimation period: 15 days
ENVIRONMENTAL CONDITIONS- Temperature (°C): 23±2°C (actual temperature: 21.9-24.1°C)- Humidity (%): 55±15% (actual humidity: 52-78%)- Air changes (per hr): 15-17- Photoperiod (hrs dark / hrs light): 12 hr dark/12 hr light (light: 8:00~20:00)

Administration / exposure
Route of administration
oral: gavage
Vehiclewater for injection
Details on oral exposure- Amount of vehicle (if gavage): 5 mL/kg- Dosing volume: 5 mL/kg
Analytical verification of doses or concentrations
yes
Details on analytical verification of doses or concentrations
Concentrations of the test solutions at the time of initial preparation were analyzed with HPLC. Ana
lytical concentrations of the test solutions were all within the range of 96.0-98.5% of the nominal
concentrations and both values were within the acceptable range.
Duration of treatment / exposure
Males: 42 days including 14 days pre-mating, 14 days mating period and 14 days after the end of themating period.
Females (mating): 42-46 days including 14 days pre-mating, mating and gestation periods and thedays until day 4 of lactation. (no mating females: 41-43 days)
Females (satellite): 42 days
Frequency of treatment
Once/day, 7 days/week
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
20

mg/kg bw/day (actual dose received)

Dose / conc.
100

mg/kg bw/day (actual dose received)

6

Dose / conc.
500

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
Mating group: 12 animals/sex /dose (0, 20, 100, and 500 mg/kg bw/day). 5 males at 0 and 500 mg/kg
bw/day were assigned to the recovery group.
Non-mating group (Satellite group): 10 females/dose (0 and 500 mg/kg bw/day). 5 females at each gr
oup were assigned to the recovery group.
Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: Based on the results of a 14-day preliminary study, the high dose was set
to 500 mg/kg bw/day, and the intermediate dose and low dose were set to 100 mg/kg bw/day and 20
mg/kg bw/day, respectively.
[14-day preliminary study]
A 14-day repeated dose oral toxicity test (Crl:CD(SD) rats, doses: 0, 100, 300, 600 or 1000 mg/kg bw/
day).
A moribund condition with dyspnea was seen in 1 male at 1000 mg/kg bw/day and 1 female at 600
mg/kg bw/day and was killed as impending. In males and females at 1000 mg/kg bw/day, irregularrespiration was observed. In males and females at 600 mg/kg bw/day and above, decrease in bodyweight and food consumption, swelling of stomach wall were observed.
- Rationale for animal assignment (if not random): Body weight-balanced randomization

Examinations
Observations and examinations performed and frequency
CAGE SIDE OBSERVATIONS: Yes- Time schedule:
2 times/day (before administration, after administration) during the administration period. Once a day
during the recovery period.
DETAILED CLINICAL OBSERVATIONS: Yes- Time schedule:
Male:
Detailed observation was conducted weekly during the administration and recovery periods.
Reactivity test, grip strength measurement, and motor activity measurement were performed once
every 6 weeks after administration.
Female:Detailed observation was conducted weekly during the administration and recovery periods. However,
only the satellite group was observed for 6 weeks after administration.
Reactivity test, grip strength measurement, and motor activity measurement were performed on the
day before anatomy in the parturition animals and once every 6 weeks in the satellite group.
BODY WEIGHT: Yes- Time schedule for examinations:
Males, females (satellite):On days 1, 8, 15, 22, 29, 36, and 42 of administration period. On days 1, 8 and 14 of recovery period.
Females (mating):
Before mating, measurements were taken on days 1, 8 and 15 of administration. Mated females were
measured at gestation 0, 7, 14, and 20. The delivered females were measured on the day of delivery
and 4 days after delivery.

7

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):- Food consumption: Yes- Time schedule for examinations:
Males, females (satellite):On days 1, 8, 15, 29, 36, 42 of administration period.
On days 1, 8, 14 of recovery period.
Females (mating):
On days 1, 8, 15 of administration period, on days 0, 7, 14, 20 of gestation period, .on days 0, 4 of lactation period.
WATER INTAKE: No
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes- Time schedule for collection of blood:
At the end of administration period, or at the end of recovery period in both sexes- Anaesthetic used for blood collection: ether- Animals fasted: Yes- How many animals:
At the end of administration period:
Males: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)Females (mating): 12 females/dose (0, 20, 100, and 500 mg/kg bw/day),Females (satellite): 5 females/dose (0 and 500 mg/kg bw/day)
At the end of recovery period:
5 males/dose and 5 females (satellite)/dose (0 and 500 mg/kg bw/day)- Parameters examined: red blood cell count, hemoglobin, hematocrit, mean corpuscular volume, mea
n corpuscular hemoglobin, mean corpuscular hemoglobin concentration, reticulocyte percentage,platelet count, white blood cell count, differential white blood cell count, prothrombin time, activated p
artial thromboplastin time.
CLINICAL CHEMISTRY: Yes- Time schedule for collection of blood:
At the end of administration period, or at the end of recovery period in both sexes- Animals fasted: Yes- How many animals:
At the end of administration period:
Males: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)Females (mating): 12 females/dose (0, 20, 100, and 500 mg/kg bw/day),Females (satellite): 5 females/dose (0 and 500 mg/kg bw/day)
At the end of recovery period:
5 males/dose and 5 females (satellite)/dose (0 and 500 mg/kg bw/day)- Parameters checked: total protein, albumin, A/G ratio, total bilirubin, glucose, total cholesterol, trigly
ceride, phospholipid, AST, ALT, LDH, ALP, γ-GT, CK, BUN, creatinine, sodium, potassium, chloride, ca
lcium, inorganic phosphorus
BLOOD HORMONE: Yes- Time schedule for collection of serum:
At the end of administration period, or at the end of recovery period in both sexes- Animals fasted: Yes- How many animals:
At the end of administration period:
Males: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)Females (mating): 12 females/dose (0, 20, 100, and 500 mg/kg bw/day),Females (satellite): 5 females/dose (0 and 500 mg/kg bw/day)
At the end of recovery period:
5 males/dose and 5 females (satellite)/dose (0 and 500 mg/kg bw/day)- Parameters checked: Triiodothyronine (T3), Thyroxin (T4), and thyroid stimulating hormone (TSH)8

URINALYSIS: Yes- Time schedule for collection of urine:
Male, female satellite group: Fresh urine was collected before administration once at week 6 ofadministration period.
- Metabolism cages used for collection of urine: No
A urine collector to collect fresh urine samples under ad libitum feeding and drinking conditions.
- How many animals:
At the end of administration period:
Male: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)Females (satellite): 10 females/dose (0 and 500 mg/kg bw/day)- Parameters checked:
Fresh urine: pH, protein, glucose, ketones, bilirubin, occult blood, urobilinogen
NEUROBEHAVIOURAL EXAMINATION: Yes- Time schedule for examinations:
Males and females (satellite): Week 6 of administration period.
Females (mating): The animals were delivered on the day before dissection (4 days after delivery).
- Dose groups that were examined: Autopsy animals after the end of the administration period- Battery of functions tested:
1) Reflex/reactions (Manipulative Test). Visual forelimb placing response, auditory reactivity, tail pinch
response, pupillary reflex, aerial righting reaction.
2) Measurement of Grip Strength. Grip strength of forelimb and hind limb was measured by MK380CM (Muromachi kikai Co.,Ltd.).
3) Measurement of Spontaneous Motor Activity. Spontaneous motor activity was measured by
SCANET MV-10 (MELQUEST Ltd.).
Motor activity was measured for 60 min.
Sacrifice and pathology
GROSS PATHOLOGY: Yes
ORGAN WEIGHT: Yes: pituitary, thyroids, heart, liver, kidney, thymus, spleen, adrenal gland, testes, epididymides, ventral
prostate, seminal vesicles (including coagulating gland) , ovaries, uterus, brain, lung
HISTOPATHOLOGY: Yes: brain. pituitary, spinal cord, eye ball, thyroid, parathyroid, heart, nasal cavity,
trachea, lung, liver, kidney, thymus, spleen, adrenal glands, stomach, small intestine (including duo
denum), large intestine, testis, epididymis, ventral prostate, seminal vesicles (including coagulating
gland), ovaries, uterus, vagina, urinary bladder, lymph nodes (axillary, inguinal, etc.), peripheral nerve(sciatic nerve), bone marrows (femur), bone (femur), mammary gland, muscle.
Statistics
For copulation index, fertility index, gestation index, urinalysis and histopathological findings, statistical difference between each treatment group and the control group was analyzed using by the C
hi-square test.
For quantitative data, homogeneity of variance was tested using Bartlett method first. If the variance
was homogenous, statistical difference between each treatment group and the control group was
analyzed using Dunnett method. If not homogenous, statistical difference between each treatment g
roup and the control group was tested using Kurskal-Wallis rank sum test and the significant differe
nce between the medium control and treated groups was analyzed by nonparametric Dunnett multiple
comparison test. For comparison of quantitative data between two groups in female satellite animals
and male recovery test, homogeneity of variance was analyzed by F-test. Then, if homogenous,
student's t-test was applied. If not homogenous, Aspin-Welch's t-test was used.

Results and discussion
9

Results of examinations
Clinical signs
effects observed, treatment-related
Description (incidence and severity)
CLINICAL SIGNS:[At the administration period]:
Abnormal respirations (males and females: respiratory noise, irregular respiration, females: oral
breathing) were observed in males and females at 500 mg/kg bw/day. Soiled perigenital region wasobserved in females at 500 mg/kg bw/day.
[At the recovery period]:
Deep respiration was observed in males at 500 mg/kg bw/day.
DETAILED CLINICAL OBSERVATIONS:[At the administration period]:
Abnormal respiration (males and females: respiratory noise, males: irregular respiration), and soiled
perinasal were observed in males and females at 500 mg/kg bw/day.
[At the recovery period]
Deep respiration, soiled perigenital region and low temperature of the skin were observed in males at
500 mg/kg bw/day.
Mortality
mortality observed, non-treatment-related
Description (incidence)
One female in the satellite group at 500 mg/kg bw/day died on day 10. In this fatal case, erosion
and inflammation in the larynx through the upper part of the trachea were observed. Obstruction in
the upper trachea by pus and congestion/edema of the lung were also observed, suggesting thatthe cause of death was suffocation in this female. To investigate obstruction in the nasal cavity, adetailed histopathological examination in the nasal mucosa of all animals was conducted; however,there were no lesions causing obstruction in any animal. An intubation error could not be excluded as
the cause of death. Therefore, this death was treatment-related but was not toxicologically relevant.
Body weight and weight changes
effects observed, treatment-related
Description (incidence and severity)[At the administration period]:
Decreased tendency body weight gain was observed in males at 500 mg/kg bw/day.
[At the recovery period]:
Decreased tendency body weight gain was observed in males at 500 mg/kg bw/day.
Food consumption and compound intake (if feeding study)no effects observed
Food efficiency
not examined
Water consumption and compound intake (if drinking water study)
not examined
Ophthalmological findings
not examined
Haematological findings
not specified
10

Description (incidence and severity)[At the end of administration period]:
Decreased MCH was observed in males at 500 mg/kg bw/day. This finding was not associated withother changes and was confined to the highest dose and no longer apparent after the recovery period. This change may be treatment related but were not necessarily adverse.
[At the end of recovery period]:
There were no changes related to the test substance in any groups.
Clinical biochemistry findings
not specified
Description (incidence and severity)
Including blood hormones (T3, T4, TSH)
CLINICAL BIOCHEMISTRY:[At the end of administration period]:
Decreased total protein and albumin were observed in males at 500 mg/kg bw/day. Decreased BUN
creatinine and IP were observed in mating females at 500 mg/kg bw/day. Decreased total protein,increased AST were observed in satellite females at 500 mg/kg bw/day.
These findings were not associated with other signs, organ weight changes, or histopathological alterations and were confined to the highest dose and no longer apparent after the recovery period.
These changes may be treatment related but were not necessarily adverse.
[At the end of recovery period]:
There were no changes related to the test substance in any groups.
BLOOD HORMONES:
There were no changes related to the test substance in any groups at the administration and recovery
periods.
Urinalysis findings
no effects observed
Behaviour (functional findings)
no effects observed
Immunological findings
not examined
Organ weight findings including organ / body weight ratios
no effects observed
Gross pathological findings
no effects observed
Neuropathological findings
not examined
Histopathological findings: non-neoplastic
no effects observed
Histopathological findings: neoplastic
not examined

Effect levels
Key result
true
11Dose descriptor
NOAEL
Effect level100

mg/kg bw/day (actual dose received)

Based ontest mat.
Sexmale/female
Basis for effect level
clinical signs
At 500 mg/kg bw/day, abnormal respiration was observed in males and females.

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
https://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF88-61-9d.pdf

Applicant's summary and conclusion
Conclusions
Based on the changes in the clinical sign, the NOAEL of repeated dose toxicity was determined to be
100 mg/kg bw/day in male and female rats.
Executive summary
In the combined repeated dose and reproductive/developmental screening test (OECD TG422), SD rats
were treated orally with the test substance at the doses of 0, 20, 100 and 500 mg/kg bw/day. Males (12animales/dose) were dosed for 42 days including a 14 day pre-mating period. Females (12 animals/dose) were dosed for 41-46 days including 14 day pre-mating, mating, and gestation periods and days
until day 4 of lactation. In addition, as the satellite study group of females (10 females/group) was dosed
0 and 500 mg/kg/day for 42 days, with 5 animals/group treated with recovery.
Irregular respiration, deep respiration, and dyspnea, which were considered to represent main toxic
effects, were observed in males and females at the 500 mg/ kg bw/day. Decreased tendency bodyweight gain was observed in males at 500 mg/kg bw/day. In the hematological examination, decreased
MCH was observed in males at 500 mg/kg bw/day. In the clinical chemistry, decreased total protein
and albumin in males, decreased BUN, creatinine and IP in mating females, decreased total protein,
and increased AST in satellite females were observed at 500 mg/kg bw/day. These findings were notassociated with other signs, organ weight changes, or histopathological alterations and were confinedto the highest dose and no longer apparent after the recovery period. These changes may be treatment
related but were not necessarily adverse.
Since respiratory abnormalities were observed at 500 mg/kg bw/day for both sexes, The NOAELs for
repeated dose toxicity of 2,4-dimethylbenzenesulfonic acid were determined to be 100 mg/kg bw/day
in male and female rats.

12

Genetic toxicity
Genetic toxicity in vitro
ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.001

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Administrative data
Endpoint
in vitro gene mutation study in bacteria
Type of information
experimental study
Adequacy of study
key study
Robust study summary
false
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study under GLP condition
Reliability 1

Data source
Reference
Reverse Mutation Test of 2,4-Dimethylbenzenesulfonic acid on Bacteria. / Ministry of Health, Labourand Welfare (MHLW), Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
13

Guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
in vitro gene mutation study in bacteria
Deviations
no
Qualifieraccording to guideline
Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
genetic toxicity in vitro, otherDeviationsno
GLP compliance
yesType of assay
bacterial reverse mutation assay
in vitro gene mutation study in bacteria

Test material
Test material information
2,4-Dimethylbenzenesulfonic acid
Specific details on test material used for the study
Purity: 99.7% (anhydrous equivalent, contained 16.1% water)

MethodSpecies / strain
Species / strain / cell typeS. typhimurium TA 1535, TA 1537, TA 98 and TA 100
bacteria
Species / strain / cell type
E. coli WP2 uvr A pKM 101
bacteria
Metabolic activation
with and without
Metabolic activation system
S9 mix: SD male rat liver, induced by phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose
-S9 mix:
313, 625, 1250, 2500, 5000 μg/plate (TA 1535, TA 1537, TA 98 and TA 100 strains)
313, 625, 1250, 2500, 5000 μg/plate (WP2uvrA/pKM101 strain)+S9 mix (10%):313, 625, 1250, 2500, 5000 μg/plate (TA 1535, TA 1537, TA 98 and TA 100 strains)14313, 625, 1250, 2500, 50000 μg/plate (WP2uvrA/pKM101 strain)+S9 mix (30%):
313, 625, 1250, 2500, 5000 μg/plate (TA 1535, TA 1537, TA 98 and TA 100 strains)
313, 625, 1250, 2500, 50000 μg/plate (WP2uvrA/pKM101 strain)
Maximum concentration was established based on the result of the preliminary test at concentration
up to 5000 ug/plate. In this test, the growth inhibition was not observed for all strains with or without
S9 mix.
Vehicle / solvent- Vehicle(s)/solvent(s) used: Distilled water
Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes
True negative controlsnoPositive controls
yes
Positive control substance
9-aminoacridine
9-amimoacridine hydrochloride (9AA): -S9 mix: (TA1537)
sodium azide
NaN3: -S9 mix: (TA1535)
furylfuramide2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (AF-2): -S9 mix: (TA100, TA98, WP2 uvrA/pkM101)
other: 2-aminoanthracene (2AA)+S9 mix: (TA1535, TA100, TA98, TA1537 and WP2 uvrA/pkM101)
Details on test system and experimental conditions
METHOD OF APPLICATION: Preincubation
DURATION- Preincubation period: 20 min at 37°C
- Exposure duration: ca.48 hrs
NUMBER OF PLATES: 3
NUMBER OF REPLICATIONS: 2
DETERMINATION OF CYTOTOXICITY- Method: other: growth inhibition
Evaluation criteria
A chemical was judged to be mutagenic when the mean number of revertant colonies per plateincreased more than twice that of the negative control and when the dose-related and reproducible increase was observed.
StatisticsnoResults and discussion
15

Test results
Key result
trueSpecies / strain
S. typhimurium TA 1535
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
True negative controls validity
not examined
Positive controls validity
valid
Key result
trueSpecies / strain
S. typhimurium TA 1537
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
True negative controls validity
not examined
Positive controls validity
valid
Key result
true
16Species / strain
S. typhimurium TA 98
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
True negative controls validity
not examined
Positive controls validity
valid
Key result
trueSpecies / strain
S. typhimurium TA 100
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
True negative controls validity
not examined
Positive controls validity
valid
Key result
trueSpecies / strain
E. coli WP2 uvr A pKM 101
bacteria
17

Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
True negative controls validity
not examined
Positive controls validityvalid

Any other information on results incl. tables
Figures and Tables (in Japanese) are available in the following full report of the study.
https://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF88-61-9e.pdf
Please also see the attached files (Tables in English)

Applicant's summary and conclusion
Conclusions
Negative with or without metabolic activation
Executive summary
In a bacterial reverse mutation assay using Salmonella typhimurium TA100, TA1535, TA98, and TA 1537,
and Escherichia coli WP2uvrA/pKM101 (OECD TG 471), 2,4-Dimethylbenzenesulfonic acid was negativewith or without metabolic activation.

18

ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.002

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Author:Date: 2022-11-14T14:54:55.000+09:00
Remarks:

Administrative data
Endpoint
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information
experimental study
Adequacy of study
key study
Robust study summary
false
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study under GLP condition
Reliability 1

Data source
Reference
In Vitro Chromosomal Aberration Test of on 2,4-Dimethylbenzenesulfonic acid Cultured Chinese
Hamster / Ministry of Health, Labour and Welfare (MHLW), Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 473 (In Vitro Mammalian Chromosomal Aberration Test)in vitro cytogenicity / chromosomal aberration study in mammalian cells (from 26 September 2014)19

Deviations
no
Qualifieraccording to guideline
Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
genetic toxicity in vitro, otherDeviationsno
GLP compliance
yesType of assay
in vitro mammalian chromosome aberration testin vitro cytogenicity / chromosome aberration study in mammalian cells

Test material
Test material information
2,4-Dimethylbenzenesulfonic acid
Specific details on test material used for the study
Purity: 99.7% (anhydrous equivalent, contained 16.1% water)

MethodSpecies / strain
Species / strain / cell type
Chinese hamster lung (CHL/IU)
mammalian cell line
Cytokinesis block (if used)colcemid
Metabolic activation
with and without
Metabolic activation system
S9 mix: SD male rat liver, induced by phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose
Cell growth inhibition study-S9 mix (short-term treatment): 0.015, 0.03, 0.059, 0.12, 0.24, 0.48, 0.95, 1.9 mg/mL+S9 mix (short-term treatment): 0.015, 0.03, 0.059, 0.12, 0.24, 0.48, 0.95, 1.9 mg/mL-S9 mix (continuous treatment, 24hr): 0.015, 0.03, 0.059, 0.12, 0.24, 0.48, 0.95, 1.9 mg/mL-S9 mix (continuous treatment, 48hr): 0.015, 0.03, 0.059, 0.12, 0.24, 0.48, 0.95, 1.9 mg/mL
Preliminary study-S9 mix (short-term treatment): 0.24, 0.48, 0.95, 1.9 mg/mL+S9 mix (short-term treatment): 0.24, 0.48, 0.95, 1.9 mg/mL-S9 mix (continuous treatment, 24hr): 0.24, 0.48, 0.95, 1.9 mg/mL-S9 mix (continuous treatment, 48hr): 0.24, 0.48, 0.95, 1.9 mg/mL
Main study-S9 (short-term treatment): 0.48, 0.95, 1.4, 1.9 mg/mL
20

+S9 (short-term treatment): 0.48, 0.95, 1.4, 1.9 mg/mL-S9 mix (continuous treatment, 24hr): 0.48, 0.95, 1.4, 1.9 mg/mL-S9 mix (continuous treatment, 48hr): 0.48, 0.95, 1.4, 1.9 mg/mL
Confirmation study+S9 (short-term treatment): 0.95, 1.4, 1.9 mg/mL

Vehicle / solvent- Vehicle(s)/solvent(s) used: Distilled water
Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes
True negative controlsnoPositive controls
yes
Positive control substancebenzo(a)pyrene+S9mitomycin C-S9Details on test system and experimental conditions
METHOD OF APPLICATION: Exposure duration: [short-term treatment]: 6 hrs + 20 hr, [continuous
treatment]: 24 hrs or 48 hrs
SPINDLE INHIBITOR: Colcemid
STAIN: Giemsa stain (2.5 v/v%) for 12 min.
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 + 100 cells /concentration
DETERMINATION OF CYTOTOXICITY- Method: relative total growth
Evaluation criteria
For the evaluation of the frequencies of structural aberrations and of polyploidy induced, the following
criteria were employed. Appearance incidence of cell with chromosomal aberrations:Negative (-): lessthan 5%, Equivocal(±): 5% or more and less than 10%, Positive(+): 10% or more
StatisticsnoResults and discussion
Test results
Key result
trueSpecies / strain
Chinese hamster lung (CHL/IU)
21

mammalian cell line
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
True negative controls validity
not examined
Positive controls validityvalid

Any other information on results incl. tables
Figures and Tables (in Japanease) are available in the following full report of the study.
https://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF88-61-9f.pdf
Please also see the attached files (Tables in English)

Applicant's summary and conclusion
Conclusions
Negative with or without metabolic activation
Executive summary
In an in vitro chromosomal aberration test using CHL/IU cells (OECD TG 473), 2,4dimethylbenzenesulfonic acid was negative with or without metabolic activation.

22

Toxicity to reproduction
Toxicity to reproduction
ENDPOINT_STUDY_RECORD: Toxicity to reproduction.001

UUID: b99d415f-750a-4ed1-a267-347b52c101ac
Dossier UUID:
Author:Date: 2022-11-29T14:06:17.424+09:00
Remarks:

Administrative data
Endpoint
reproductive toxicity, other Combined Repeated Dose Toxicity Study with the Reproduction /Developmental Toxicity Screening Test
Type of information
experimental study
Adequacy of study
key study
Robust study summary
false
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
other: OECD Test Guideline study under GLP condition
Cross-reference
Reason / purpose for cross-reference
reference to same study 7.5.1 Repeated dose toxicity: oral: Repeated dose toxicity: oral. 001
Related information
OECD / Repeated dose toxicity: oral / Repeated dose toxicity: oral.001 / 2,4-Dimethylbenzenesulfonicacid / 2,4-dimethylbenzenesulfonic acid / 88-61-9

Data source
Reference
Combined repeated dose toxicity study with the reproductive/developmental toxicity screening test of /Ministry of Health, Labour and Welfare (MHLW), Japan / study report
Data accessdata published

Materials and methods
23Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction /Developmental Toxicity Screening Test)
Deviations
yes During the power outage (October 2, 2010) due to the inspection of the electrical equipmen
t of the facility, the humidity increased (78%), the lights were turned off, and the ventilation was
stopped.
However, no effect was observed on the test results.
GLP compliance
yes
Limit test
no

Test material
Test material information
2,4-Dimethylbenzenesulfonic acid
Specific details on test material used for the study- Name of test material (as cited in study report): 2,4-Dimethylbenzenesulfonic acid
- Analytical purity: 99.7% (anhydrous equivalent, contained 16.1% water)- Storage condition of test material: Room temperature, shading- Stability under test conditions: The stability of test material was identified by analysis of the remainder.

Test animals
Speciesrat
Strainother: Crl:CD(SD)Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source: Charles River Japan, Inc., Atsugi Breeding Center.
- Age at study initiation: 10 weeks old- Weight at study initiation: Male: 355-428 g, Female: 220-249 g- Housing:
Mating period: Stainless steel mesh cages for group housing (340W × 294D × 176H mm)
After 18 days of pregnancy: Stainless steel cage for delivery (340W × 294D × 176H mm) and bedding.
Other breeding period: 2-piece stainless steel mesh cage (170W × 294D × 176H mm/animal)- Diet: Solid feed (CRF-1: Oriental Yeast Co., ltd.) was given ad libitum.
- Water: Tap water was given ad libitum.
- Acclimation period: 15 days
24

ENVIRONMENTAL CONDITIONS- Temperature (°C): 23±2°C (actual temperature: 21.9-24.1°C)- Humidity (%): 55±15% (actual humidity: 52-78%)- Air changes (per hr): 15-17- Photoperiod (hrs dark / hrs light): 12 hr dark/12 hr light (light: 8:00~20:00)

Administration / exposure
Route of administration
oral: gavage
Vehiclewater for injection
Details on exposure- Amount of vehicle (if gavage): 5 mL/kg- Dosing volume: 5 mL/kg
Details on mating procedure- M/F ratio per cage:1/1- Length of cohabitation: up to 14 days
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
Analytical verification of doses or concentrations
yes
Details on analytical verification of doses or concentrations
Concentrations of the test solutions at the time of initial preparation were analyzed with HPLC. Ana
lytical concentrations of the test solutions were all within the range of 96.0-98.5% of the nominal
concentrations and both values were within the acceptable range.

Duration of treatment / exposure
Males: 42 days including 14 days pre-mating, 14 days mating period and 14 days after the end of themating period.
Females (mating): 42-46 days including 14 days pre-mating, mating and gestation periods and thedays until day 4 of lactation. (no mating females: 41-43 days)
Females (satellite): 42 days
Frequency of treatment
Once/day, 7 days/week
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
20

mg/kg bw/day (actual dose received)25

Dose / conc.
100

mg/kg bw/day (actual dose received)

Dose / conc.
500

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
Mating group: 12 animals/sex /dose (0, 20, 100, and 500 mg/kg bw/day). 5 males at 0 and 500 mg/kg
bw/day were assigned to the recovery group.
Non-mating group (Satellite group): 10 females/dose (0 and 500 mg/kg bw/day). 5 females at each gr
oup were assigned to the recovery group.
Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: Based on the results of a 14-day preliminary study, the high dose was set
to 500 mg/kg bw/day, and the intermediate dose and low dose were set to 100 mg/kg bw/day and 20
mg/kg bw/day, respectively.
[14-day preliminary study]
A 14-day repeated dose oral toxicity test (Crl:CD(SD) rats, doses: 0, 100, 300, 600 or 1000 mg/kg bw/
day).
A moribund condition with dyspnea was seen in 1 male at 1000 mg/kg bw/day and 1 female at 600
mg/kg bw/day and was killed as impending. In males and females at 1000 mg/kg bw/day, irregularrespiration was observed. In males and females at 600 mg/kg bw/day and above, decrease in bodyweight and food consumption, swelling of stomach wall were observed.
- Rationale for animal assignment (if not random): Body weight-balanced randomization

Examinations
Parental animals: Observations and examinations
CAGE SIDE OBSERVATIONS: Yes- Time schedule:
2 times/day (before administration, after administration) during the administration period. Once a day
during the recovery period.
DETAILED CLINICAL OBSERVATIONS: Yes- Time schedule:
Male:
Detailed observation was conducted weekly during the administration and recovery periods.
Reactivity test, grip strength measurement, and motor activity measurement were performed once
every 6 weeks after administration.
Female:Detailed observation was conducted weekly during the administration and recovery periods. However,
only the satellite group was observed for 6 weeks after administration.
Reactivity test, grip strength measurement, and motor activity measurement were performed on the
day before anatomy in the parturition animals and once every 6 weeks in the satellite group.
BODY WEIGHT: Yes- Time schedule for examinations:
Males, females (satellite):On days 1, 8, 15, 22, 29, 36, and 42 of administration period. On days 1, 8 and 14 of recovery period.
26

Females (mating):
Before mating, measurements were taken on days 1, 8 and 15 of administration. Mated females were
measured at gestation 0, 7, 14, and 20. The delivered females were measured on the day of delivery
and 4 days after delivery.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):- Food consumption: Yes- Time schedule for examinations:
Males, females (satellite):On days 1, 8, 15, 29, 36, 42 of administration period.
On days 1, 8, 14 of recovery period.
Females (mating):
On days 1, 8, 15 of administration period, on days 0, 7, 14, 20 of gestation period, .on days 0, 4 of lactation period.
WATER INTAKE: No
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes- Time schedule for collection of blood:
At the end of administration period, or at the end of recovery period in both sexes- Anaesthetic used for blood collection: ether- Animals fasted: Yes- How many animals:
At the end of administration period:
Males: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)Females (mating): 12 females/dose (0, 20, 100, and 500 mg/kg bw/day),Females (satellite): 5 females/dose (0 and 500 mg/kg bw/day)
At the end of recovery period:
5 males/dose and 5 females (satellite)/dose (0 and 500 mg/kg bw/day)- Parameters examined: red blood cell count, hemoglobin, hematocrit, mean corpuscular volume, mea
n corpuscular hemoglobin, mean corpuscular hemoglobin concentration, reticulocyte percentage,platelet count, white blood cell count, differential white blood cell count, prothrombin time, activated p
artial thromboplastin time.
CLINICAL BIOCHEMISTRY: Yes- Time schedule for collection of blood:
At the end of administration period, or at the end of recovery period in both sexes- Animals fasted: Yes- How many animals:
At the end of administration period:
Males: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)Females (mating): 12 females/dose (0, 20, 100, and 500 mg/kg bw/day),Females (satellite): 5 females/dose (0 and 500 mg/kg bw/day)
At the end of recovery period:
5 males/dose and 5 females (satellite)/dose (0 and 500 mg/kg bw/day)- Parameters checked: total protein, albumin, A/G ratio, total bilirubin, glucose, total cholesterol, tri
glyceride, phospholipid, AST, ALT, LDH, ALP, γ-GT, CK, BUN, creatinine, sodium, potassium, chloride,
calcium, inorganic phosphorus
BLOOD HORMONE: Yes- Time schedule for collection of serum:
At the end of administration period, or at the end of recovery period in both sexes- Animals fasted: Yes- How many animals:
At the end of administration period:
Males: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)27

Females (mating): 12 females/dose (0, 20, 100, and 500 mg/kg bw/day),Females (satellite): 5 females/dose (0 and 500 mg/kg bw/day)
At the end of recovery period:
5 males/dose and 5 females (satellite)/dose (0 and 500 mg/kg bw/day)- Parameters checked: Triiodothyronine (T3), Thyroxin (T4), and thyroid stimulating hormone (TSH)
URINALYSIS: Yes- Time schedule for collection of urine:
Male, female satellite group: Fresh urine was collected before administration once at week 6 ofadministration period.
- Metabolism cages used for collection of urine: No
A urine collector to collect fresh urine samples under ad libitum feeding and drinking conditions.
- How many animals:
At the end of administration period:
Male: 7, 12, 12, and 7 males/dose (0, 20, 100, and 500 mg/kg bw/day)Females (satellite): 10 females/dose (0 and 500 mg/kg bw/day)- Parameters checked:
Fresh urine: pH, protein, glucose, ketones, bilirubin, occult blood, urobilinogen
NEUROBEHAVIOURAL EXAMINATION: Yes- Time schedule for examinations:
Males and females (satellite): Week 6 of administration period.
Females (mating): The animals were delivered on the day before dissection (4 days after delivery).
- Dose groups that were examined: Autopsy animals after the end of the administration period- Battery of functions tested:
1) Reflex/reactions (Manipulative Test). Visual forelimb placing response, auditory reactivity, tail pinch
response, pupillary reflex, aerial righting reaction.
2) Measurement of Grip Strength. Grip strength of forelimb and hind limb was measured by
MK-380CM (Muromachi kikai Co.,Ltd.).
3) Measurement of Spontaneous Motor Activity. Spontaneous motor activity was measured by SCAN
ET MV-10 (MELQUEST Ltd.).
Motor activity was measured for 60 min.
Oestrous cyclicity (parental animals)
Vaginal smears were collected from all females except satellites and microscopically examined every
day from the day after the start of administration until the day copulation was confirmed.
Sperm parameters (parental animals)
Parameters examined in all P male parental generations: testes, epididymis ventral prostate, and sem
inal vesicle weight, histopathological examinations for testes, epididymis ventral prostate, and sem
inal vesicle.
Litter observations
PARAMETERS EXAMINED: The following parameters were examined in F1 offspring: Number and sexof pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, and weight gain.
GROSS EXAMINATION OF DEAD PUPS: Yes, for external and internal abnormalities.
Postmortem examinations (parental animals)
METHOD OF SACRIFICED: All animals were sacrificed by exsanguination under ether anesthesia.
SACRIFICE: Males, females (satellite): On next day after the last administration, Maternal animals: on
Day 5 of lactation, Females who copulated but did not deliver: The day corresponding to GD 26, Males
and females recovery group: on Day 14 of recovery.
GROSS PATHOLOGY: Yes
ORGAN WEIGHT: Yes
28

: pituitary, thyroids, heart, liver, kidney, thymus, spleen, adrenal gland, testes, epididymides, ventral
prostate, seminal vesicles (including coagulating gland) , ovaries, uterus, brain, lung
HISTOPATHOLOGY: Yes: brain. pituitary, spinal cord, eye ball, thyroid, parathyroid, heart, nasal cavity,
trachea, lung, liver, kidney, thymus, spleen, adrenal glands, stomach, small intestine (including duodenum), large intestine, testis, epididymis, ventral prostate, seminal vesicles (including coagulating
gland), ovaries, uterus, vagina, urinary bladder, lymph nodes (axillary, inguinal, etc.), peripheral nerve (sciatic nerve), bone marrows (femur), bone (femur), mammary gland, muscle.
Postmortem examinations (offspring)
SACRIFICE- The F1 offsprings were euthanized on PND4 by exsanguination under ether anesthesia.
GROSS NECROPSY: Yes- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, andabdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGTHS- Not examined.
Statistics
For copulation index, fertility index, gestation index, urinalysis and histopathological findings, statistical difference between each treatment group and the control group was analyzed using by the C
hi-square test.
For quantitative data, homogeneity of variance was tested using Bartlett method first. If the variance
was homogenous, statistical difference between each treatment group and the control group was
analyzed using Dunnett method. If not homogenous, statistical difference between each treatment g
roup and the control group was tested using Kurskal-Wallis rank sum test and the significant differe
nce between the medium control and treated groups was analyzed by nonparametric Dunnett multiple
comparison test. For comparison of quantitative data between two groups in female satellite animals
and male recovery test, homogeneity of variance was analyzed by F-test. Then, if homogenous,
student's t-test was applied. If not homogenous, Aspin-Welch's t-test was used.
Reproductive indices
Each parameter was determined by the following equations:
Copulation index (%) = (No. of copulated pairs / No. of mated pairs) × 100
Fertility index (%) = (No. of pregnant females / No. of copulated pairs) × 100
Gestation index (%) = (No. of females with complete parturition / No. of pregnant females) × 100
Gestation length (days)
Implantation index (%) = (No. of implantations / No. of corpora lutea) × 100
Delivery index (%) = (No. of pups born / No. of implantation) × 100
Birth index (%) = (No. of live pups on day 0 / No. of implantation) × 100
Live birth index (%) = (No. of live pups on day 0 / No. of pups born) × 100
Sex ratio on pups born = No. of male pups / No. of male and female pups born
Sex ratio on day 4 = No. of male live pups on day 4 / No. of live pups on day 4
Offspring viability indices
Viability index (%) = (No. of live pups on day 4 / No. of live pups on day 0) × 100

Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)29

Clinical signs
effects observed, treatment-related
Description (incidence and severity)
See 7.5.1 Repeated dose toxicity. 001
Mortality
mortality observed, non-treatment-related
Description (incidence)See 7.5.1 Repeated dose toxicity. 001
Body weight and weight changes
effects observed, treatment-related
Description (incidence and severity)
See 7.5.1 Repeated dose toxicity. 001
Food consumption and compound intake (if feeding study)no effects observed
Food efficiency
not examined
Water consumption and compound intake (if drinking water study)
not examined
Ophthalmological findings
not examined
Haematological findings
not specified
Description (incidence and severity)
See 7.5.1
Clinical biochemistry findings
not specified
Description (incidence and severity)
See 7.5.1 Repeated dose toxicity. 001
Urinalysis findings
no effects observed
Behaviour (functional findings)
no effects observed
Immunological findings
not examined
Organ weight findings including organ / body weight ratios
no effects observed
Gross pathological findings
no effects observed
Neuropathological findings
not examined
Histopathological findings: non-neoplastic
no effects observed30Histopathological findings: neoplastic
not examined

Reproductive function / performance (P0)
Reproductive function: oestrous cycle
no effects observed
Reproductive function: sperm measures
no effects observed
Reproductive performanceno effects observed

Details on results (P0)General toxicity: See 7.5.1 Repeated dose toxicity. 001
Reproductive function / performance: There were no effects on reproductive parameters up to 500
mg/kg bw/day.

Effect levels (P0)
Key result
true
Dose descriptor
NOAEL
Effect level500

mg/kg bw/day (actual dose received)

Based ontest mat.
Sexmale/female
Basis for effect level
other: No effects on reproduction

Results: F1 generation
General toxicity (F1)Clinical signs
no effects observed
Mortality / viability
no mortality observed
Body weight and weight changes
no effects observed
Gross pathological findings
no effects observed31

Details on results (F1)
There were no effects on developmental parameters up to 500 mg/kg bw/day.

Effect levels (F1)
Key result
true
Dose descriptor
NOAEL
Generation
F1
Effect level500

mg/kg bw/day (actual dose received)

Based ontest mat.
Sexmale/female
Basis for effect level
other: No effects on development

Overall reproductive toxicity
Key result
true
Reproductive effects observednoAny other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
https://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF88-61-9d.pdf

Applicant's summary and conclusion
Conclusions
In the combined repeated oral dose toxicity study with the reproduction/developmental toxicity screening test (OECD TG 422) described above, there were no effects on reproductive and developmental
parameters up to 500 mg/kg bw/day. The NOAEL for the rat reproductive/developmental toxicity of
2,4-Dimethylbenzenesulfonic acid was regarded as 500 mg/kg bw/day, the highest dose tested.
Executive summary
In the combined repeated dose and reproductive/developmental screening test (OECD TG422), SD rats
were treated orally with the test substance at the doses of 0, 20, 100 and 500 mg/kg bw/day. Males were32

dosed for 42 days including a 14 day pre-mating period. Females (12 animals/dose) were dosed for
41-46 days including 14 day pre-mating, mating, and gestation periods and days until day 4 of lactation.
In addition, as the satellite study group of females (10 females/group) was dosed 0 and 500 mg/kg/
day for 42 days, with 5 animals/group treated with recovery. There were no effects on reproductive anddevelopmental parameters up to 500 mg/kg bw/day. The NOAEL for the rat reproductive/developmental
toxicity of 2,4-dimethylbenzenesulfonic acid was regarded as 500 mg/kg bw/day, the highest dosetested.

33

References
Reference Substances
REFERENCE_SUBSTANCE: m-xylene-4-sulphonic acid
UUID: ECB5-25d98114-4c1a-4381-882e-fa63892577f7
Dossier UUID:
Author:Date: 2007-05-10T18:00:00.000+09:00
Remarks:
Reference substance namem-xylene-4-sulphonic acid
IUPAC name
2,4-dimethylbenzenesulfonic acidInventory
Inventory number
Inventory namem-xylene-4-sulphonic acidInventory
EC Inventory
Inventory number
201-843-9
CAS number
88-61-9
Molecular formulaC8H10O3S
Description
CAS number
88-61-9

Synonyms
Synonyms
Identity
Benzenesulfonic acid, 2,4-dimethylIdentity
Benzenesulfonic acid, 2,4-dimethyl-

Molecular and structural information
34

Molecular formulaC8H10O3S
Molecular weight186.2282
SMILES notation
Cc1ccc(c(C)c1)S(=O)(=O)O
InChl
InChI=1/C8H10O3S/c1-6-3-4-8(7(2)5-6)12(9,10)11/h3-5H,1-2H3,(H,9,10,11)
Structural formula

Related substancesGroup / category information
DSL Category: Organics

35

Test Materials
TEST_MATERIAL_INFORMATION: 2,4Dimethylbenzenesulfonic acid
UUID: 889d6bfe-5432-4110-bc28-9bf04b35ad65
Dossier UUID:
Author:Date: 2021-03-04T09:50:09.000+09:00
Remarks:
Name
2,4-Dimethylbenzenesulfonic acid36

Literatures
LITERATURE: Combined repeated dose toxicity study withthe reproductive/developmental toxicity screening test of
2,4-Dimethylbenzenesulfonic acid by oral administrationin rats
UUID: 5c5e4887-b688-420d-a735-389d5aa77069
Dossier UUID:
Author:Date: 2021-03-04T09:35:01.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Combined repeated dose toxicity study with the reproductive/developmental toxicity screening test of
2,4-Dimethylbenzenesulfonic acid by oral administration in rats
Author
Ministry of Health, Labour and Welfare (MHLW), Japan
Bibliographic source
available in the web of Japan Existing Chemical Data Base (JECDB)
https://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF88-61-9d.pdf
Testing facility
Japan Bioassay Research Center
Report number
0765

37

LITERATURE: In Vitro Chromosomal Aberration Test of
on 2,4-Dimethylbenzenesulfonic acid Cultured Chinese
Hamster Cells.
UUID: 54541599-236b-4c5c-b2aa-7afdcd5e9032
Dossier UUID:
Author:Date: 2021-03-15T16:17:50.000+09:00
Remarks:

General information
Reference Typestudy report
Title
In Vitro Chromosomal Aberration Test of on 2,4-Dimethylbenzenesulfonic acid Cultured Chinese
Hamster Cells.
Author
Ministry of Health, Labour and Welfare (MHLW), Japan
Year
2011
Bibliographic source
Japan Existing Chemical Data Base (JECDB)
https://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF88-61-9f.pdf
Testing facility
Japan Bioassay Research Center
Report number
7416

38

LITERATURE: Reverse Mutation Test of 2,4Dimethylbenzenesulfonic acid on Bacteria.
UUID: 478e3901-a792-4655-941f-4bcb69910f50
Dossier UUID:
Author:Date: 2021-03-10T15:31:49.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Reverse Mutation Test of 2,4-Dimethylbenzenesulfonic acid on Bacteria.
Author
Ministry of Health, Labour and Welfare (MHLW), Japan
Year
2011
Bibliographic source
Japan Existing Chemical Data Base (JECDB)
https://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF88-61-9e.pdf
Testing facility
Japan Bioassay Research Center
Report number
6337

39

Legal Entities
LEGAL_ENTITY: National Institute of Health Sciences
UUID: IUC4-b036ff75-0f3c-323b-b200-ed5f46cf5101
Dossier UUID:
Author:Date: 2022-11-07T15:49:29.000+09:00
Remarks:

General information
Legal entity nameNational Institute of Health Sciences
Remarks
Disclaimer: The contents in this document were created based on the MHLW (Ministry of Health,Labour and Welfare) peer reviewed study reports (in Japanese) in JECDB (Japan Existing Chemical
Database) at http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp. Authorship is in the Divisionof Risk Assessment, the National Institute of Health Sciences, and the contents do not reflect any official MHLW opinions or any other regulatory policies.

Address
Address 1
Tonomachi 3-25-26
Address 2
Kawasaki-ku
Postal code
210-9501
Town
KawasakiRegion / State
Kanagawa
CountryJapan
JP

Identifiers
Other IT system identifiers
IT system
LEO
ID
10767
IT systemIUCLID440

ID16558402024DIV750

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