Name: COMPLETE / SUBSTANCE : 1,2,4,5-Benzenetetracarboxylic acid / benzene-1,2,4,5tetracarboxylic acid / 89-05-4 Mon, 12 Dec 2022, 09:41:47+0900 /Legal entity owner: National Institute of Health Sciences
Printing date: 2022-12-12T09:41:47.355+09:00

iTable of Contents
0/0 ............................................................................................................................. 1
National Institute of Health Science ..................................................................... 2
1,2,4,5-Benzenetetracarboxylic acid ...................................................................... 3
CORE .......................................................................................................... 3
1 General information .......................................................................... 3
1.10 Assessment approach (assessment entities) ......................... 3
Assessment approach (assessment entities) ........................ 3
OECD .......................................................................................................... 4
D Health Effects .................................................................................. 4
67 Repeated dose toxicity: oral .................................................... 4
Repeated dose toxicity: oral.001 ........................................... 470 Genetic toxicity in vitro ......................................................... 11
Genetic toxicity in vitro.001 ................................................ 11
Genetic toxicity in vitro.002 ................................................ 1773 Toxicity to reproduction ........................................................ 21
Toxicity to reproduction.001 ............................................... 21
DOMAIN .................................................................................................... 28
Substance ......................................................................................... 28
Substance ................................................................................ 28
References ................................................................................................................ 29
Reference Substances ...................................................................................... 29
benzene-1,2,4,5-tetracarboxylic acid ............................................................ 29
Test Materials .................................................................................................. 31
1,2,4,5-Benzenetetracarboxylic Acid ............................................................ 31
Literatures ....................................................................................................... 32
Combined Repeated Dose Toxicity Study with the Reproduction/
Developmental Toxicity Screening Test of 1,2,4,5-Benzenetetracarboxylic
acid by Oral Administration in Rats ............................................................. 32
In Vitro Chromosomal Aberration Test of on 1,2,4,5-benzenetetracarboxylic
acid Cultured Chinese Hamster Cells. ......................................................... 33
Reverse Mutation Test of 1,2,4,5-Benzenetetracarboxylic Acid on
Bacteria. ................................................................................................... 34
Legal Entities ................................................................................................... 35
National Institute of Health Sciences .......................................................... 35ii

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1,2,4,5-Benzenetetracarboxylic acid / benzene-1,2,4,5-tetracarboxylic acid / 89-05-4
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National Institute of Health Science
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1

LEGAL_ENTITY: National Institute of Health
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General information
Legal entity nameNational Institute of Health Science2

1,2,4,5-Benzenetetracarboxylic acid
CORE
General information
Assessment approach (assessment entities)
FIXED_RECORD: Assessment approach

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Dossier UUID:
Author:Date: 2018-02-27T15:49:57.000+09:00
Remarks:3

OECD
Health Effects
Repeated dose toxicity: oral
ENDPOINT_STUDY_RECORD: Repeated dose toxicity: oral.001

UUID: 5d2623ad-aba5-430b-867e-16a36feba0ab
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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
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Testof / MHLW (Ministry of Health, Labour and Welfare), Japan / study report
Data accessdata published

Materials and methods
4Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction /Developmental Toxicity Screening Test)
GLP compliance
yes
Limit test
no

Test material
Test material information
1,2,4,5-Benzenetetracarboxylic Acid
Specific details on test material used for the study
1,2,4,5-Benzenetetracarboxylic Acid;Purity 99.9% (CAS:89-05-4)

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 (Atsugi)- Age at study initiation:
10 weeks old- Weight at study initiation:
342 g -400 g (male); 213 g - 260 g (female)- Fasting period before study:- Housing:metal cage (W250 x D350 x H200 mm), (W340 x D400 x H185 mm for pregnant animals on GD17 to
PND 5)- Diet:ad libitum- Water:ad libitum- Acclimation period:
14 days
DETAILS OF FOOD AND WATER QUALITY:ENVIRONMENTAL CONDITIONS- Temperature (°C):
520-26- Humidity (%):
30-60
- Air changes (per hr):12
- Photoperiod (hrs dark / hrs light):
19:00-7:00/7:00-19:00

Administration / exposure
Route of administration
oral: gavage
Vehiclemethylcellulose
Details on oral exposure
Vehicle: 0.5 w/v% methylcellulose solution
Analytical verification of doses or concentrations
yes
Duration of treatment / exposure
males: 42 days, females: 41-46 days from 14 days before mating to day 4 of lactation
Frequency of treatment
once a day
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
100

mg/kg bw/day (actual dose received)

Dose / conc.
300

mg/kg bw/day (actual dose received)

Dose / conc.
1000

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
17(12 animals as an administration group and 5 animals as a recovery group) /sex/dose (0 and 1000
mg/kg bw/day)
12/sex/dose (100 and 300 mg/kg/day)6

Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale:
Dose finding study: Rats were dosed the test substance for 14 days at 0, 100, 300 and 1000 mg/kg
bw/day. Soft feces were observed at 1000 mg/kg bw/day in both sexes, but no effects were observed
on body weight. The highest dose was set at 1000 mg/kg bw/day for the main study.
- Post-exposure recovery period in satellite groups: 14 days

Examinations
Observations and examinations performed and frequency
Clinical observation performed and frequency: General condition was observed 3 times a day during
the administration period (before dosing, and immediately after and approximately 2 hours after
dosing) and once a day (in the morning) during the recovery period.
Detailed clinical observation was done for all animals. It was done once a week during the admin
istration period and recovery period for males and non-mated females, and once a week during the
pre-mating administration period and on the designated days during the mating, gestation andlactation periods for females in the mating group (on days 1, 7, 14 and 20 of gestation for the copu
lated females and on day 4 of lactation for the females that delivered). As detailed clinical obser
vations, the animals were observed for the following items: posture, convulsion and abnormal behavio
r in the home cage observation; ease of removal from cage, reactivity to handling (ease of handling,
vocalization, etc.) condition of fur and skin (staining of fur, unkempt fur, injury, color of skin, etc.),
eyeball (exophthalmos, palpebral closure), secretions from eyes and nose, mucosal membranes,
autonomic nervous function (lacrimation, salivation, piloerection, pupil size and respiration) to handling at in-the-hand observation; and arousal, gait, posture, tremor, convulsion, rearing count, defecation (defecation count, urination), stereotypy (grooming, circling, etc.), abnormal behavior (selfbiting, backward walking, etc.) in the open field observation.
Manipulative test and measurement of grip strength and motor activity were done for 5 animals ineach group: males in the main groups were examined in the final week of administration, females in
the main groups on day 4 of lactation, and males and females in the recovery groups in the final weekof administration and in the final week of recovery. Animals were examined for auditory response,
approach response, touch response, tail pinch response, pulillary reflex, aerial righting reflex and land
ing foot splay. The grip strength of the forelimbs and hind limbs was measured. The motor activity
was measured for 1 hour and measured values of 10-minute intervals and 0-60 minute value wererecorded.
Body weights were determined on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 of administration for males and on days 1, 4, 8, 11 and 15 of administration, days 0, 4, 7, 11, 14, 17 and 20 of gestationand days 0 and 4 of lactation for females, and the day of necropsy in males and females. In addition,
males and females in the recovery groups were weighed on days 1, 4, 8, 11 and 14 of recovery and on
the day of necropsy.
Food consumption was determined on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration formales and on days 1, 4, 8, 11 and 15 of administration, days 1, 4, 7, 11, 14, 17 and 20 of gestationand days 2 and 4 of lactation in females, but it was not determined during the mating period for mal
es and females. In addition, it was determined on days 1, 4, 8, 11 and 14 of recovery for males andfemales in the recovery groups
Urinalysis was done for 5 males in each group and 5 non-mated group females in the final week ofadministration and in the final week of recovery.
In all animals in the control group and the high dose group and 5 males and 5 females in the low andmiddle dose groups, hematological examination and blood chemistry examination were carried out attime of necropsy after the end of administration or recovery period.

7

Sacrifice and pathology
Necropsy: Detailed macroscopic examination was conducted on the organs/tissues throughout the
body of each animal, including the external appearance, head, thorax and abdomen.
Measurement of organ weighs: The brain, thyroids (including parathyroids), adrenals, thymus, spleen,
heart, liver, kidneys, testes, epididymides were determined.
Histopathological examination: The stomach in males and females of all groups, the cerebrum,
cerebellum, pituitary, spinal cord (thoracic), sciatic nerve, thyroids, parathyroids, adrenals, thymus,
spleen, submandibular lymph node, mesenteric lymph node, heart, lung (including bronchus), stomach, duodenum, jejunum, ileum, cecum, colon, rectum, liver, kidneys, urinary bladder, testes, epidid
ymides, ovaries, uterus, seminal vesicles, sternum (including bone marrow), femur (including bone marrow) in males and females at 0 and 1000 mg/kg bw/day.
Statistics
Dunnett's test for continuous data, Dunnett-type mean rank test for quantal data and chi-square testwith Yates’ continuity correction or chi-square test with Yates’ continuity correction for other data
were used.

Results and discussion
Results of examinations
Clinical signs
effects observed, treatment-related
Description (incidence and severity)
soft feces were observed in males and females in the 1000 mg/kg bw/day group.
Mortality
no mortality observed
Body weight and weight changes
no effects observed
Food consumption and compound intake (if feeding study)no effects observed
Haematological findings
no effects observed
Clinical biochemistry findings
no effects observed
Urinalysis findings
no effects observed
Behaviour (functional findings)
no effects observed
Organ weight findings including organ / body weight ratios
no effects observed
Gross pathological findings
no effects observed
Histopathological findings: non-neoplastic
effects observed, treatment-related8Description (incidence and severity)
At the end of the administration period, squamous hyperplasia at the limiting was observed in stomac
h of males in the 1000 mg/kg group. However, reversibility was observed for this lesion at the end of
the recovery period.

Effect levels
Key result
false
Dose descriptor
NOAEL
Effect level300

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
histopathology: non-neoplastic

Target system / organ toxicity
Key result
false
Critical effects observed
yes
Lowest effective dose / conc.
1000

mg/kg bw/day (nominal)

Systemgastrointestinal tract
Organ
intestine
stomachTreatment related
yes
Dose response relationshipyes

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
9http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF89-05-4b.pdf

Applicant's summary and conclusion
Conclusions
Based on the effects in the gastrointestinal tract, the NOAEL for local effects on rat regarding therepeated-dose toxicity of 1,2,4,5-benzenetetracarboxylic acid was determined to be 300 mg/kg bw/
day.
Executive summary
A combined repeated oral dose toxicity study with the reproduction/developmental toxicity screening
test was performed as described in OECD TG 422. Male and female rats (12 animals/sex/dose) wereadministered 1,2,4,5-benzenetetracarboxylic acid at 0 (vehicle:0.5 w/v% methyl cellulose solution), 100,
300, and 1,000 mg/kg bw/day. Males were dosed for 42 days, including a 14-day pre-mating period andsubsequent mating period, whereas females were dosed for 41–46 days, including 14-day pre-mating,mating, and gestation periods, and until lactation day 4. Five out of 12 males administered 1,2,4,5benzenetetracarboxylic acid at 0 and 1,000 mg/kg bw/day were treated as a recovery group and examined
after a 14-day recovery period. Regarding the findings of clinical observation, soft feces were observed inmales and females of the 1,000 mg/kg bw/day group. Upon histopathological examination, hyperplasia of
the squamous epithelium at the limiting ridge, considered to be due to irritation by the test substance, wasfound in the stomach of males of the 1,000 mg/kg bw/day group at the end of the administration period.
Reversibility was observed for this lesion at the end of the recovery period. Based on the effects in thegastrointestinal tract, the NOAEL for local effects on rat regarding the repeated-dose toxicity of 1,2,4,5benzenetetracarboxylic acid was determined to be 300 mg/kg bw/day.

10

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

UUID: 40c20b28-8c31-4ea3-8153-d63404543a5b
Dossier UUID:
Author:Date: 2022-12-12T09:36:49.556+09:00
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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
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Reverse Mutation Test of 1,2,4,5-Benzenetetracarboxylic Acid on Bacteria. / Ministry of Health, Labourand Welfare (MHLW), Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
in vitro gene mutation study in bacteria

11

GLP compliance
yesType of assay
bacterial reverse mutation assay
in vitro gene mutation study in bacteria

Test material
Test material information
1,2,4,5-Benzenetetracarboxylic Acid
Specific details on test material used for the study
1,2,4,5-Benzenetetracarboxylic Acid;Purity 99.9% (CAS:89-05-4)

MethodSpecies / strain
Species / strain / cell typeS. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2bacteria
Metabolic activation
with and without
Metabolic activation system
S9 mix: Rat liver, induced with phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose
Dosage of each strain with or without S9-S9 mix: 0, 313, 625, 1250, 2500, 5000 ug/plate(all strains)+S9 mix: 0, 156, 313, 625, 1250, 2500, 5000 ug/plate(TA strains)
0, 313, 625, 1250, 2500, 5000 ug/plate (WP2 uvrA)
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 observed at 5000 μg/plate for S. typhimu
rium TA100, TA1535, TA98 and TA1537 with S9 mix.
Vehicle / solvent
Dimethylsulfoxide
Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes
True negative controlsnoPositive controls
yes
Positive control substancesodium azide
12without S9 mix:(TA 1535)
benzo(a)pyrene
with S9 mix: (TA100, TA98, TA1537)
other:
without S9 mix:2-(2-Furyl)-3-(5-nitro -2-furyl)acrylamide (TA100, TA98, WP2uvrA), ICR-191(TA1537) with S9 mix: 2-Aminoanthracene (TA1535, WP2 uvrA)
Details on test system and experimental conditions
METHOD OF APPLICATION: Preincubation
DURATION- Preincubation period: 20 min at 37°C
- Exposure duration: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.
Statistics
not used

Results and discussion
Test results
Key result
false
Species / strain
S. typhimurium TA 100
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity at 5000 μg/plate (+S9 mix)
Vehicle controls validity
valid
Untreated negative controls validity
not examined
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 1535
13

bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity at 5000 μg/plate (+S9 mix)
Vehicle controls validity
valid
Untreated negative controls validity
not examined
Positive controls validity
valid
Key result
false
Species / strain
E. coli WP2 uvr A pKM 101
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Untreated negative controls validity
not examined
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 98
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity at 5000 μg/plate (+/- S9 mix)
14

Vehicle controls validity
valid
Untreated negative controls validity
not examined
Positive controls validity
valid
Key result
false
Species / strain
S. typhimurium TA 1537
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity at 5000 μg/plate (+S9 mix)
Vehicle controls validity
valid
Untreated negative controls validity
not examined
Positive controls validity
valid
Additional information on results
There were no precipitation in any test concentration.

Any other information on results incl. tables
Figures and Tables (in Japanese) are available in the following full report of the study. http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF89-05-4e.pdf
Tables (in English) are attachted to this document. Please download the export file to see the Tables.

Applicant's summary and conclusion
Conclusions
Genotoxic effects:
With metabolic activation: Negative
Without metabolic activation: Negative
Executive summary
In a bacterial reverse mutation assay usingS. typhimuriumTA100, TA1535, TA98, and TA1537, andE.
coliWP2uvrA/pKM101 (OECD TG 471), negative results were obtained for 1,2,4,5-benzenetetracarboxylic
acid with or without metabolic activation.
15

16

ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.002

UUID: a11081cc-e5df-404c-92c0-f44363410538
Dossier UUID:
Author:Date: 2019-09-03T11:22:59.000+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
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
In Vitro Chromosomal Aberration Test of on 1,2,4,5-benzenetetracarboxylic acid Cultured Chinese Ham /MHLW, Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)17

in vitro cytogenicity / chromosome aberration study in mammalian cells
Qualifieraccording to guideline
Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
genetic toxicity in vitro, other
GLP compliance
yesType of assay
in vitro mammalian chromosome aberration testin vitro cytogenicity / chromosome aberration study in mammalian cells

Test material
Test material information
1,2,4,5-Benzenetetracarboxylic Acid
Specific details on test material used for the study
1,2,4,5-Benzenetetracarboxylic Acid;Purity 99.9% (CAS:89-05-4)

MethodSpecies / strain
Species / strain / cell typeother: Chinese hamster lung(CHL/IU) cell
Metabolic activation
with and without
Metabolic activation system
S9 mix: Rat liver, induced with phenobarbital and 5,6- benzoflavone
Test concentrations with justification for top dose
-S9 mix(short-term treatment): 0, 676.8, 947.5, 1327, 1857, 2600 μg/mL+S9 mix(short-term treatment): 0, 750, 900, 1050, 1200, 1500, 1650 μg/mL-S9 mix(24hr-continuous treatment): 0, 900, 1050, 1200, 1500, 1650, 1800 μg/mL-S9 mix(48hr-continuous treatment): 0, 1000, 1100, 1200, 1300, 1400 μg/m
Cell-growth inhibition test was conducted up to the limited concentration of 2600 μg/mL (10 mM)-Short term treatment, +S9 mix: concentration of 50% cell-groth inhibition was determined as 2523.5
μg/mL-Short term treatment, -S9 mix: concentration of 50% cell-groth inhibition was determined as 2321.4
μg/mL-Continous treatment (24 h): concentration of 50% cell-groth inhibition was determined as 2600.0 μg/
mL-Continous treatment (48 h): concentration of 50% cell-groth inhibition was determined as 1246.9 μg/
mL
Vehicle / solvent
DMSO
18

Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes
True negative controlsnoPositive controls
yes
Positive control substancecyclophosphamide(with S9)
mitomycin C(without S9)
Details on test system and experimental conditions
METHOD OF APPLICATION: Exposure duration: [short-term treatment]:6 hrs + 18 hr, [continuous
treatment]: 24h, 48 h
Plates/test: 2
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
Statistics
Not used

Results and discussion
Test results
Key result
false
Species / strainother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
Positive controls validity
valid

19

Any other information on results incl. tables
1,2,4,5-Benzenetetracarboxylic acid did not induce polyploidy under the conditions of this study, but apositive result in the incidence of the occurrence of chromosome structural aberrations was reproduced,
though it was not dose-dependent. Since remarkable lowering in the pH of the culture was observed ,it was judged that the structural aberrations induced by the test article were caused by the culturalenvironment and thus non-specific.

Genetic effects:

Clastogenicity

Polyploidy

+ ? -

+ ? -

Without metabolic activation[ ] [ ] [*][ ] [ ] [*]With metabolic activation[ ] [ ] [*][ ] [ ] [*]24hr-continuous treatment[*] [ ] [ ][ ] [ ] [*]clastogenicity is considered to be
due to low pH condition

Figures and Tables (in English) are available in the following full report of the study.
http://dra 4.nihs.go.jp/mhlw_data/home/pdf/PDF89-05-4f.pdf

Applicant's summary and conclusion
Executive summary
In anin vitrochromosomal aberration test using CHL/IU cells (OECD TG 473), negative results wereobtained with or without metabolic activation.

20

Toxicity to reproduction
ENDPOINT_STUDY_RECORD: Toxicity to reproduction.001

UUID: ec21741c-15da-4df3-865b-f757841e8f8a
Dossier UUID:
Author:Date: 2022-12-12T09:35:30.729+09:00
Remarks:

Administrative data
Endpoint
screening for reproductive / developmental toxicity
Type of information
experimental study
Adequacy of study
key study
Robust study summary
true
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1

Data source
Reference
Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Testof / MHLW (Ministry of Health, Labour and Welfare), Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction /Developmental Toxicity Screening Test)

21

GLP compliance
yes

Test material
Test material information
1,2,4,5-Benzenetetracarboxylic Acid
Specific details on test material used for the study
1,2,4,5-Benzenetetracarboxylic Acid;Purity 99.9% (CAS:89-05-4)

Test animals
Speciesrat
Strainother: Crl:CD(SD)Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source:
Charles river (Atsugi)- Age at study initiation:
10 weeks old- Weight at study initiation:
342 g -400 g (male); 213 g - 260 g (female)- Fasting period before study:- Housing:metal cage (W250 x D350 x H200 mm), (W340 x D400 x H185 mm for pregnant animals on GD17 to
PND 5)- Diet:ad libitum- Water:ad libitum- Acclimation period:
14 days
DETAILS OF FOOD AND WATER QUALITY:ENVIRONMENTAL CONDITIONS- Temperature (°C):
20-26- Humidity (%):
30-60
- Air changes (per hr):12
- Photoperiod (hrs dark / hrs light):
19:00-7:00/7:00-19:00

Administration / exposure
Route of administration
oral: gavage
22

Vehicleother: 0.5 w/v% methylcellulose solution
Details on mating procedure
Method of mating: Males and females in the same dose group of the main groups were co-housed
overnight on a one-to-one basis after the end of the pre-mating administration period. Copulation
was considered successful if the formation of vaginal plugs or presence of sperm in vaginal smears
was confirmed the following morning. The length of the mating period for the same male andfemale was 5 days at maximum.
Analytical verification of doses or concentrations
yes
Duration of treatment / exposure
males: 42 days, females: 41-46 days from 14 days before mating to day 4 of lactation
Frequency of treatment
once a day
Doses / concentrations
Dose / conc.
0mg/kg bw/day (actual dose received)

Dose / conc.
100

mg/kg bw/day (actual dose received)

Dose / conc.
300

mg/kg bw/day (actual dose received)

Dose / conc.
1000

mg/kg bw/day (actual dose received)

No. of animals per sex per dose
17(12 animals as an administration group and 5 animals as a recovery group) /sex/dose (0 and 1000
mg/kg bw/day)
12/sex/dose (100 and 300 mg/kg/day)Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale:
23

Dose finding study: Rats were dosed the test substance for 14 days at 0, 100, 300 and 1000 mg/kg
bw/day. Soft feces were observed at 1000 mg/kg bw/day in both sexes, but no effects were observed
on body weight. The highest dose was set at 1000 mg/kg bw/day for the main study.
- Post-exposure recovery period in satellite groups: 14 days

Examinations
Parental animals: Observations and examinations
Clinical observation performed and frequency: General condition was observed 3 times a day during
the administration period (before dosing, and immediately after and approximately 2 hours after
dosing) and once a day (in the morning) during the recovery period.
Body weights were determined on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 of adminis
tration for males and on days 1, 4, 8, 11 and 15 of administration, days 0, 4, 7, 11, 14, 17 and 20 of
gestation and days 0 and 4 of lactation for females, and the day of necropsy in males and females. In addition, males and females in the recovery groups were weighed on days 1, 4, 8, 11 and 14 ofrecovery and on the day of necropsy.
Food consumption was determined on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration for males and on days 1, 4, 8, 11 and 15 of administration, days 1, 4, 7, 11, 14, 17 and 20 of gestation and days
2 and 4 of lactation in females, but it was not determined during the mating period for males andfemales. In addition, it was determined on days 1, 4, 8, 11 and 14 of recovery for males and females
in the recovery groups.
Oestrous cyclicity (parental animals)
Vaginal smears were collected from all females in the main groups and microscopically examined
every day (in the morning) from the day after the start of administration until the day copulation was
confirmed. Vaginal smear pictures were classified as proestrus, estrus, metestrus and diestrus andexamined for the frequency of estrus and interval between estruses (estrous cycle).
Litter observations
Examination of liveborn pups: The numbers of liveborn pups and stillborn pups were counted on the
day of birth. After liveborn pups were examined for any external abnormality, sexed and weighed,dams were allowed to nurse their pups. Liveborn pups were observed for mortality once daily until
day 4 after birth. All liveborn pups were exsanguinated after measurement of body weight on day
4 after birth, necropsied and examined for any abnormality in organs/tissues, including those in thehead, thorax and abdomen. Individual body weights of liveborn pups were recorded, and the average
body weight per litter was calculated by sex.
Pathological examinations were performed.
Postmortem examinations (parental animals)
See 7.5.1 Repeated dose toxicity
Postmortem examinations (offspring)
GROSS NECROPSY- Gross necropsy consisted of external examination
Statistics
Dunnett's test for continuous data, Dunnett-type mean rank test for quantal data and chi-square testwith Yates’ continuity correction or chi-square test with Yates’ continuity correction for other data
were used.
Reproductive indicesNo. of copulated animals, No. of males that impregnated females, No. of pregnant females, No. offemales that delivered liveborn pups, estrous cycle, gestational length, No.of corpora lutea, No. of im
plantation sites, total No. of liveborn and stillborn pups
Offspring viability indicesNo. of liveborn pups, sex ratio on day 0 and day 4 after birth, copulation index (No. of copulated animals / No. of animals housed together x 100), insemination index (No. of pregnant females / No.
24

of copulated males x 100), fertility index (No. of pregnant females / No. of copulated females x100), delivery index (No. of females that delivered liveborn pups / No. of pregnant females x 100),
implantation index (No. of implantation sites / No. of corpora lutea x100), stillbirth index (No. of stillborn pups / No. of pups born x 100), index of external abnormalities (No. of pups with external abn
ormalities / No. of pups born x 100), live birth index (No. of liveborn pups / No. of pups born x 100),and viability index on day 4 after birth (No. of live pups on day 4 after birth / No. of liveborn pups x
100)

Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
Clinical signs
effects observed, treatment-related
Mortality
no mortality observed
Body weight and weight changes
no effects observed
Food consumption and compound intake (if feeding study)no effects observed
Haematological findings
no effects observed
Clinical biochemistry findings
no effects observed
Urinalysis findings
no effects observed
Behaviour (functional findings)
no effects observed
Organ weight findings including organ / body weight ratios
no effects observed
Gross pathological findings
no effects observed
Histopathological findings: non-neoplastic
effects observed, treatment-related

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

Effect levels (P0)25Key result
true
Dose descriptor
NOAEL
Effect level1000

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
reproductive performanceNo effects observed
Key result
false
Dose descriptor
NOAEL
Effect level300

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect level
clinical signs
soft feces were observed in males and females in the 1000 mg/kg grouphistopathology: non-neoplasticsquamous hyperplasia at the limiting was observed in stomach of males in the 1000 mg/kg bw/
daygroup

Results: F1 generation
General toxicity (F1)Mortality / viability
no mortality observed
Body weight and weight changes
no effects observed

Effect levels (F1)26

Key result
false
Dose descriptor
NOAEL
Generation
F1
Effect level1000

mg/kg bw/day (actual dose received)

Based onact. ingr.
Sexmale/female
Basis for effect levelviability
no effects
body weight and weight gainno effects

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF89-05-4b.pdf

Applicant's summary and conclusion
Executive summary
A combined repeated oral dose toxicity study with the reproduction/developmental toxicity screening
test was performed as described in OECD TG 422. Male and female rats (12 animals/sex/dose) wereadministered 1,2,4,5-benzenetetracarboxylic acid at 0 (vehicle:0.5 w/v% methyl cellulose solution), 100,
300, and 1,000 mg/kg bw/day. Males were dosed for 42 days, including a 14-day pre-mating period
and subsequent mating period, whereas females were dosed for 41–46 days, including 14-day premating, mating, and gestation periods, and until lactation day 4. Five out of 12 males administered
1,2,4,5-benzenetetracarboxylic acid at 0 and 1,000 mg/kg bw/day were treated as a recovery group andexamined after a 14-day recovery period. Regarding the findings of clinical observation, soft feces wereobserved in males and females of the 1,000 mg/kg bw/day group. Upon histopathological examination,
hyperplasia of the squamous epithelium at the limiting ridge, considered to be due to irritation by thetest substance, was found in the stomach of males of the 1,000 mg/kg bw/day group at the end of theadministration period. Reversibility was observed for this lesion at the end of the recovery period. There
were no effects on reproductive and developmental parameters at 1,000 mg/kg bw/day. The NOAEL forthe rat reproductive/developmental toxicity of 1,2,4,5-benzenetetracarboxylic acid was thus regarded
as 1,000 mg/kg bw/day, the highest dose tested.

27

DOMAIN
Substance
SUBSTANCE: 1,2,4,5-Benzenetetracarboxylic acid
UUID: 916619f2-21b5-4bc8-8261-6a1c647257b2
Dossier UUID:
Author:Date: 2022-12-12T09:36:49.556+09:00
Remarks:
Substance name
1,2,4,5-Benzenetetracarboxylic acid
Legal entity
National Institute of Health Sciences / Kawasaki / Japan

Identification of substance
Reference substancebenzene-1,2,4,5-tetracarboxylic acid / benzene-1,2,4,5-tetracarboxylic acid / 89-05-4 / 201-879-5
EC number

EC name201-879-5

EC Inventory

CAS number

CAS name

89-05-4
IUPAC namebenzene-1,2,4,5-tetracarboxylic acid

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

28

References
Reference Substances
REFERENCE_SUBSTANCE: benzene-1,2,4,5tetracarboxylic acid
UUID: ECB5-b71b5f8a-c0f0-46d8-91ea-ab7fbd98c99d
Dossier UUID:
Author:Date: 2007-05-10T18:00:00.000+09:00
Remarks:
Reference substance namebenzene-1,2,4,5-tetracarboxylic acid
IUPAC namebenzene-1,2,4,5-tetracarboxylic acidInventory
Inventory number
Inventory namebenzene-1,2,4,5-tetracarboxylic acidInventory
EC Inventory
Inventory number
201-879-5
CAS number
89-05-4
Molecular formula
C10H6O8
Description
CAS number
89-05-4

Synonyms
Synonyms
Identity
1,2,4,5-Benzenetetracarboxylic acid

Molecular and structural information
29

Molecular formula
C10H6O8
Molecular weight254.1498
SMILES notation
OC(=O)c1cc(C(=O)O)c(cc1C(=O)O)C(=O)O
InChl
InChI=1/C10H6O8/c11-7(12)3-1-4(8(13)14)6(10(17)18)2-5(3)9(15)16/h1-2H,(H,11,12)(H,13,14)(H,15,16)(H,17,18)
Structural formula30Test Materials
TEST_MATERIAL_INFORMATION: 1,2,4,5Benzenetetracarboxylic Acid
UUID: 7ae2f323-f6d3-4dd8-90b0-50035b807a11
Dossier UUID:
Author:Date: 2018-03-02T16:28:06.000+09:00
Remarks:
Name
1,2,4,5-Benzenetetracarboxylic Acid

31

Literatures
LITERATURE: Combined Repeated Dose Toxicity Studywith the Reproduction/Developmental Toxicity Screening
Test of 1,2,4,5-Benzenetetracarboxylic acid by Oral
Administration in RatsUUID: 89f9517b-c78d-4b09-bd20-f94f1315025c
Dossier UUID:
Author:Date: 2018-03-06T09:17:24.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening
Test of 1,2,4,5-Benzenetetracarboxylic acid by Oral Administration in Rats
AuthorMHLW (Ministry of Health, Labour and Welfare), Japan
Year
2009
Bibliographic source
Japan Existing Chemical Data Base (JECDB) http://dra4.nihs.go.jp/mhlw_data/jsp/Sea rchPage
ENG.jsp
Testing facility
Gotemba Laboratory, Bozo Research Center Inc. Gotemba Shizuoka

32

LITERATURE: In Vitro Chromosomal Aberration Test of
on 1,2,4,5-benzenetetracarboxylic acid Cultured Chinese
Hamster Cells.
UUID: 9de6c7ea-59f8-4ab2-8ba7-809fb6fed7a2
Dossier UUID:
Author:Date: 2018-03-08T15:09:43.000+09:00
Remarks:

General information
Reference Typestudy report
Title
In Vitro Chromosomal Aberration Test of on 1,2,4,5-benzenetetracarboxylic acid Cultured Chinese
Hamster Cells.
Author
MHLW, Japan
Year
2008
Bibliographic source
Japan Existing Chemical Data Base (JECDB) http://dra4.nihs.go.jp/mhlw_data/jsp/Sea rchPage
ENG.jsp
Testing facility
Bozo Research Center Inc.

33

LITERATURE: Reverse Mutation Test of 1,2,4,5Benzenetetracarboxylic Acid on Bacteria.
UUID: 28e216ef-11ab-4f22-a9e6-05cf1412f7c8
Dossier UUID:
Author:Date: 2018-08-27T11:46:23.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Reverse Mutation Test of 1,2,4,5-Benzenetetracarboxylic Acid on Bacteria.
Author
Ministry of Health, Labour and Welfare (MHLW), Japan
Year
2007
Bibliographic source
JECDB http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp
Testing facility
Bozo Research Center Inc.

34

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 systemIUCLID435

ID16558402024DIV750

36