Name: COMPLETE / SUBSTANCE : 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5triazinane-2,4,6-trithione / 638-16-4 Fri, 16 Dec 2022, 11:14:48+0900 /Legal entity owner: National Institute of Health Sciences
Printing date: 2022-12-16T11:14:48.380+09:00

iTable of Contents
0/0 ............................................................................................................................. 1
National Institute of Health Science ..................................................................... 2
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione .................................................................. 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
60 Acute toxicity: oral .................................................................. 4
Acute toxicity: oral.001 ........................................................ 4
67 Repeated dose toxicity: oral .................................................... 8
Repeated dose toxicity: oral.001 ........................................... 870 Genetic toxicity in vitro ......................................................... 17
Genetic toxicity in vitro.001 ................................................ 17
Genetic toxicity in vitro.002 ................................................ 2171 Genetic toxicity in vivo .......................................................... 25
Genetic toxicity in vivo.001 ................................................ 25
73 Toxicity to reproduction ........................................................ 30
Toxicity to reproduction.001 ............................................... 30
DOMAIN .................................................................................................... 37
Substance ......................................................................................... 37
Substance ................................................................................ 37
References ................................................................................................................ 38
Reference Substances ...................................................................................... 38
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione ........................................................ 38
Test Materials .................................................................................................. 40
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8 ................................... 40
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8 ................................... 41
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8 ................................... 42
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8 ................................... 43
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8 ................................... 44
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8 ................................... 45
Literatures ....................................................................................................... 46
A combined repeated-dose/reproductive-developmental toxicity study oftrithiocyanuric acid by oral administration in rats. ........................................ 46
In Vitro Chromosomal Aberration Test of Trithiocyanuric acid on Cultured
Chinese Hamster Cells. ............................................................................. 47
Micronucleus test of Trithiocyanuric acid on mouse .................................... 48
Reverse Mutation Test of trithiocyanuric acid on Bacteria. ........................... 49
Single Dose Oral Toxicity Test of Trithiocyanuric Acid in Rats ....................... 50
Legal Entities ................................................................................................... 51
National Institute of Health Sciences .......................................................... 51

ii

DOSSIER:UUID: 0
Dossier UUID:
Author:Date: 2022-12-16T11:14:48.122+09:00
Remarks:

Dossier header
Dossier submission type
Name
Complete table of contents
Versioncore 7.0
Name (given by user)

Dossier subject
Dossier subject
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4
Public nameSubmitting legal entity
National Institute of Health Science
Dossier creation date/time
Fri, 16 Dec 2022, 11:14:48+0900
Used in category

1

LEGAL_ENTITY: National Institute of Health
Science
UUID: f51e7b54-9211-4863-90ce-fcf8a155d647
Dossier UUID:
Author:Date: 2022-11-07T16:24:02.822+09:00
Remarks:

General information
Legal entity nameNational Institute of Health Science2

1,3,5-triazine-2,4,6(1H,3H,5H)-trithione
CORE
General information
Assessment approach (assessment entities)
FIXED_RECORD: Assessment approach

UUID: ef98227b-ca5b-3c1c-a2a3-65ccb25e850d
Dossier UUID:
Author:Date: 2017-01-04T16:32:27.000+09:00
Remarks:3

OECD
Health Effects
Acute toxicity: oral
ENDPOINT_STUDY_RECORD: Acute toxicity: oral.001

UUID: IUC5-ee8931c1-a3d9-42f4-b95f-e4564603c53f
Dossier UUID:
Author:Date: 2022-12-14T15:52:59.296+09:00
Remarks:

Administrative data
Endpoint
acute 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

Data source
Reference
Single Dose Oral Toxicity Test of Trithiocyanuric Acid in Rats / MHW (Ministry of Health and Welfare),Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
4Guideline
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations
no
Qualifieraccording to guideline
Guideline
other: Yakushoku-hatsu 1121003, Kanpoki-hatsu 031121004
GLP compliance
yes
Test type
acute toxic class method
Limit test
no

Test material
Test material information
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Test animals
Speciesratcommon species
Strainother: Crj: CD(SD), SPF
SexfemaleDetails on test animals or test system and environmental conditions
TEST ANIMALS- Source: Charles River Japan Inc.
- Age at the time of purchase: 7 weeks old- Weight at dosing: 186.2 – 202.4 g (1st dosing) , 204.2-219.6 g (2nd dosing), 227.2-242.2 g (3rddosing)– Used animal number: A total of 18 females- Fasting period before study: Approximately 16 hrs- Housing: One animal/cage- Diet (e.g. ad libitum): Ad libitum except fasting period for 16 hrs before dosing to 3 hrs after dosing- Water (e.g. ad libitum): Ad libitum- Acclimation period: 8 days.
ENVIRONMENTAL CONDITIONS- Temperature (°C): 23.0 – 24.5
- Humidity (%): 49.0 – 59.0
- Ventilation (per hr): Approximately 15 times- Photoperiod (hrs light / hrs dark): 12/12

Administration / exposure
5Route of administration
oral: gavage
Vehicleother: sodium carboxymethyl cellulose
Details on oral exposure
Concentration in vehicle: 0.05, 0.5, 3, and 20% (w/v).
Doses
300 mg/kg bw (first and second dosing)2000 mg/kg bw (third dosing)No. of animals per sex per doseFirst and second dosing (first purchase): each 3 females (animal ID No. 1 – 3 and 4 - 6)/300 mg/kg b
w doseThird dosing (second purchase): 3 females (animal ID No. 7 – 9) /2000 mg/kg bw dose
Control animals
noDetails on study design- Duration of observation period following dosing: 14 days
- Frequency of observations: Before dosing, Day 1 (day of dosing): Continuously observed until 1 hr,thereafter, every one hour until 6 hours. After day 2: once a day- Frequency of weighing: Just before dosing ( Day 1), Day 2, 4, 8, 11, and 15. And when the deadanimals found- Necropsy of survivors performed: Yes
Statistics
No

Results and discussion
Effect levels
Key result
false
Sex
female
Dose descriptor
LD50
Effect level
>

300

2000

mg/kg bw

Based onact. ingr.
Key result
false
Sex
female6

Dose descriptor
LD50
Effect level500

mg/kg bw

Based onact. ingr.
Remarks on result
other: Cut off value
Mortality
No deaths were observed from first and second dosing (300 mg/kg bw). All 3 animals died in 2000
mg/kg bw group (third dosing).
Clinical signs
other: One animal received 2000 mg/kg bw of the test substance showed creeping, lid closure, lateralposition and dyspnea and died at approximately 2 hours from the dosing. Smudge of perinasal area,
listless and reduction o(defecation were observed in two other
Gross pathology
At necropsy, while no marked abnormalities were observed in the animals treated with 300 mg/kg bw,
enlargements of the kidney and the adrenal gland, and atrophy of the spleen were observed in two
animals died on Day 3 or 4 of the observation period with 2,000 mg/kg bw. White-colored liquid in thestomach was observed in two animals died at 2 hours and 3 days after dosing.
Other findings- Organ weights: No data
- Histopathology: No lesions attributable to the test substance were observed in the 300 mg/kg bw
groups. In histological examination, all the animals treated with 2000 mg/kg bw showed necrosis and degenerations of the proximal tubular epithelium and the glomerulus of the renal cortex, and two
animals showed a decrease in the area of white pulp of the spleen.
- Potential target organs: Kidneys- Other observations: No data

Applicant's summary and conclusion
Conclusions
The LD50 value was 300 – 2000 mg/kg bw , and the cut off LD50 value was 500 mg/kg bw for female
rats.

7

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

UUID: IUC5-dcf7dfcd-9eeb-434e-bda2-c5418425112e
Dossier UUID:
Author:Date: 2022-12-16T11:10:56.374+09:00
Remarks:

Administrative data
Endpoint
short-term repeated dose toxicity: oral combined repeated dose and reproduction / developmental
screening
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
Remarks
7.8.1 Toxicity to reproduction: Toxicity to reproduction.001

Data source
Reference
A combined repeated-dose/reproductive-developmental toxicity study of trithiocyanuric acid by oral a /MHW (Ministry of Health and Welfare), Japan / study report
Data accessdata published8

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

Test material
Test material information
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Test animals
Speciesratcommon rodent species
Strainother: Crj: CD(SD), SPF
Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source: Atsugi Breeding Center, Charles River Laboratories Japan, Inc.
- Age at study initiation: 8 weeks of age- Weight at study initiation: 355.5 – 405.2 g for males and 208.7 – 255.0 g for females- Housing: bracket-type metallic wire-mesh cages (W 220 × D 270 × H 190 mm)- Diet (e.g. ad libitum): ad libitum- Water (e.g. ad libitum): ad libitum- Acclimation period: 14 days
ENVIRONMENTAL CONDITIONS- Temperature (°C): 22.5 to 26.0°C- Humidity (%): 50.0 to 65.0%- Air changes (per hr): 15 times per hour
- Photoperiod (hrs dark / hrs light):12-hour lighting per day

Administration / exposure
Route of administration
oral: gavage
9Vehicleother: sodium carboxymethyl cellulose
Details on oral exposure
VEHICLE- Justification for use and choice of vehicle: No data
- Amount of vehicle (if gavage): 5 ml/kg bw- Supplier: Maruishi pharmaceutical industry Co., Ltd.
- Lot/batch no. (if required): 4720
- Dosing volume: 5 mL/kg- Stability (test solutions): For 8 days
- Storage condition of test solution: Stored in a dark place with room temperature
Analytical verification of doses or concentrations
yes
Details on analytical verification of doses or concentrations
Test suspensions at each concentration (20 and 0.05 w/v%) of initial and final preparations wereanalyzed by the HPLC method at Hatano laboratory of Food and Drug Safety Center. Results show
ed that the concentration of the test article in each concentration was 94.3 to 101% of the nominal
concentration.
Duration of treatment / exposure(P) Males: 42 days including 14 days pre-mating(P)Females: Days including 14 days pre-mating, mating and gestation periods and the days until day
4 of lactation Female (no mating, satellite group): for 42 days
Frequency of treatment
Daily: 7 times / week
Doses / concentrations
Remarks
Doses / Concentrations:
0 (vehicle), 62.5, 125 and 250 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose
12/sex/dose (main group), and 5 females/dose at 0 and 250 mg/kg bw/day (satellite group)
Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: A preliminary study (study No. R-04-006) was conducted to determine thedoses to be employed. Male and female rats were receiving 0, 250, 500, and 1000 mg/kg bw/day for
14 days. As a result, red colored urine was observed in more than half numbers of rats receiving 1000
mg/kg bw/day groups and all animals died in the 1000 mg/kg bw/day groups until Day 7. Black pathzones of auricle and dark purple colored tail top were observed in the 500 mg/kg bw/day groups from
Week 2, and moreover, one female sowing red colored urine, salivation, wasting and sedation were
moribund . Body weight gains were depressed in male rats receiving 500 mg/kg bw/day at Week 2,
and swelling of the spleen was observed at necropsy. No changes related to the test substance wereobserved in males receiving 250 mg/kg bw/day. From the results of the preliminary study, 500 mg/kg
bw/day seemed to be the maximum tolerance dose for 14 days. Therefore, the high dose was set at
250 mg/kg bw/day for the main study, and the middle and low doses were set at 125 and 62.5 mg/kg
bw/day using common ratio 2.
10

Positive controlnoExaminations
Observations and examinations performed and frequency
CAGE SIDE OBSERVATIONS: Yes- Time schedule:
Males and females: once before the start of dosing, two times/day during the dosing period, and once
during the recovery period
DETAILED CLINICAL OBSERVATIONS: Yes- Time schedule:
Male animals: the end of acclimation period, Day 7, 14, 21, 28, 35 and 42 during the dosing and Day 7
and 14 during the recovery period. Female animals: the end of acclimation period, Day 7, 14, 21, 28,
35 and 42 during the dosing, Day 0 at delivery period and Day 0 from 4 at delivery animals, and Day
7 and 14 of satellite groups during the recovery period. Test items are following: Body position, loc
omotor activity, vocalization, tremor, convulsion by cage side observation, ease of removing rat fromcage, reactivity to being handled, heart rate, body temperature, fur, skin color, visible mucosa color,
lacrimation, exophthalmos ophthalmocele, pupillary, and salivation by terminology for removing rat
from cage, and body position, walking, grooming, phonation, straub tail, gait, stereotyped, abnormal
behavior, tremor, convulsion, piloerection, and ophthalmorrhexis by observation of behavior on theworking table.
FANCTIONAL OBSERVATION: Yes- Time schedule:
End of the dosing period: male, female (dam), and female (satellite group) animals. End of the dosing
period: male and female (satellite group) animals.
Test items are following: Prayer’s reaction, pupillary reflex, visual placing, startle reaction, withdrawal
reflex, eyelid reflex, and righting reflex.
BODY WEIGHT: Yes
Males in the main and females in satellite groups were weighed on Day 1, 7, 14, 21, 28, 35 and 42 of
dosing, and Day 1, 7, and 15 (necropsy day) at the end of the recovery period.
Females were weighed on Day 1, 7, 14, and 21 until successful copulation. Copulated females were
weighed on Day 0, 7, 14 and 20 of gestation, days 0 and 4 of lactation, and the necropsy day.
FOOD CONSUMPTION : Yes- Food consumption (g/day/rat) for each animal determined from the difference of the of the previous
day's feeding amount: Yes
Measurement of food consumption was conducted on all animals at the following frequencies:
Males and females in the satellite groups: on Day 1-2, 7-8, 14-15, 29-30, 35-36, and 41-42 during thedosing period and Day 6-7 and 13-14 during the recovery period. Females in the main groups: on Day
1-2, 7-8, and 14-15 (before mating), on Day 0-1, 7-8, 14-15, and 20-21 (the gestation period). On Day
3-4 (the lactation period).
FOOD INTAKE: No
HAEMATOLOGY: Yes- Time schedule for collection of blood: On Day 43 of the dosing period and Day 15 of the recovery period in males. On Day 5 of the lactation period (main group) and on Day 15 of the recovery period (satellite group) in females.
- Anaesthetic used for blood collection: Yes (pentobarbital sodium)- Animals fasted: Yes (for 18-22 hours)- How many animals: 5 animals/sex/group- Parameters examined red blood cell count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, reticulocyte count, platelet
count, white blood cell count, differential white blood cell count, prothrombin time, activated partial
thromboplastin time
CLINICAL CHEMISTRY: Yes- Time schedule for collection of blood: Same as hematology examination.
- Animals fasted: Yes (for 18-22 hours)- How many animals: 5 animals/sex/group11

- Parameters checked: ASAT (GOT), ALAT (GPT), γ-GT, ALP, total bilirubin, blood urea nitrogen, crea
tinine, glucose, total cholesterol, triglyceride, total protein, albumin, A/G ratio, calcium, inorganic
phosphorus, sodium, potassium, chloride
URINALYSIS OF MALES: Yes- Time schedule for collection of urine: Five males on Day 33 during the dosing period and all animals
on Day 13 during the recovery period.
Five females on Day 34 (main group). All animals on Day 34 during the dosing period and Day 13 dur
ing the recovery period (satellite group).
- Metabolism cages used for collection of urine: Yes- Animals fasted: Yes- Parameters checked: color, muddy, pH, occult blood, protein, glucose, ketones. bilirubin, urobilino
gen, and sediments
BLOOD HORMONE: No
NEUROBEHAVIOURAL EXAMINATION: Yes
Detailed clinical observation: on Day 7, 14, 21, 28, 35, and 42 of the dosing period, on Day 7 and 14 of
the recovery period,
Functions test: on Day 42 of the dosing period and on Day 14 of the recovery period (males and satell
ite group females). On Day 4 of the lactation period (main group females).
Sacrifice and pathology
GROSS PATHOLOGY AND ORGAN WEIGHTS：Yes Brain, heart, liver, kidneys, adrenals, thymus, spleen, testes, and epididymis.
HISTOPATHOLOGY: Yes Brain, puitality, thymus, lymph nodes (including mesenteric and mandibul
ar lymph nodes), trachea, lung (including bronchus), stomach, intestinal tract (duodenum, jejunum, ileum, cecum, colon, rectum), thyroids, heart, liver, spleen, kidneys, adrenals, urinary bladder, testes,
epididymis, seminal vesicles (including the coagulating gland), prostate (ventral lobe), ovaries, uterus,
vagina, bone marrow (one side femur), sciatic nerve (one side femur), spinal cord, and gross abnorm
alities site.
Statistics
Changes in estrous cyclicity and conception rate were analyzed by Fisher’s test. Graded pathological
data was analyzed by Mann-Whitney’s U test (significance level = 0.05) and pathological data with number of positive and negative animals was analyzed by one-sided Fisher’s test.
Other data, obtained values in each animal or mean of a litter was one data, and these data were compared among the satellite groups and other among the groups. These data were analyzed using Ftest for homogeneity of distribution. The Student’s t-test and the Aspin-Welch’s t-test were conducted
for homogenous and non-homogenous distribution, respectively to compare the control and individual treatment groups. Three or more groups setting, these data were analyzed using Bartlett’s testfor homogeneity of distribution. The Dunnett’s multiple comparison test after the ANOVA and the Du
nnett’s-type mean rank sum test after Kruskal-Wallis’s H test were conducted for homogenous andnon-homogenous distribution, respectively to compare the control and individual treatment groups.
Significance level was set at 0.05 compared with the control group and among the groups.

Results and discussion
Results of examinations
Clinical signs
effects observed, treatment-related
Mortality
mortality observed, treatment-related
Body weight and weight changes
effects observed, treatment-related
12

Food consumption and compound intake (if feeding study)
effects observed, treatment-related
Haematological findings
effects observed, treatment-related
Clinical biochemistry findings
effects observed, treatment-related
Urinalysis findings
effects observed, treatment-related
Organ weight findings including organ / body weight ratios
effects observed, treatment-related
Gross pathological findings
effects observed, treatment-related
Histopathological findings: non-neoplastic
effects observed, treatment-related
Details on results
CLINICAL SIGNS AND MORTALITY: Mortality: One male and one female died in the 250 mg/kg bw/
day group on Day 8 and Day 42, respectively. Clinical signs: Incomplete eyelid opening and emaciatio
n were observed in one dead female receiving 250 mg/kg bw/day, but no clinical sign was observedin one dead male receiving 250 mg/kg bw/day. The followings were observed in the 250 mg/kg bw/day groups: black area of the pinna in one male and three females, dark purple distally of the tail in
two males, nodule of the tail in four males and one female, nodule of the pinna and emaciation in
one male and one female, anemic in one male, induction of the scrotum in one male, nodule of thescrotum in three males, swelling of the left forelimb in one male, and crust formation of the snout in
one female. Reddish urine was observed in one male receiving 125 mg/kg bw/day and three males
and one female receiving 250 mg/kg bw/day groups. Sporadically salivation just after dosing wasobserved in six males receiving 125 mg/kg bw/day and eight males and 10 females receiving 250
mg/kg bw/day groups. Loss of fur was observed in two males in the control group, one male and one
female receiving 125 mg/kg bw/day, and one female receiving 250 mg/kg bw/day. Crust formation of
the dorsal neck was observed in one female receiving 62.5 mg/kg bw/day and mass of the perineal
region was observed in one female receiving 125 mg/kg bw/day. No adverse changes were observedin both sexes during the recovery period.
DETAILED CLINICAL OBSERVATIONS AND FANCTIONAL OBSERVATION: Detailed clinical observa
tions, no changes were observed in both sexes during the recovery period. When removing rats fromthe cage or handling rats, vocalization and flinches were observed in both sexes sporadically, and running around in the cage was observed in one female rat receiving 250 mg/kg bw/day. When handlingrats, tenseness or rigidity was observed in one female rat receiving 125 mg/kg bw/day and 250 mg/
kg bw/day. Rearing of stereotypy was observed in two females receiving 250 mg/kg bw/day.
Functional observation, no changes were observed in both sexes during the dosing and recovery per
iods.
BODY WEIGHT: Significant decreases in body weights, weight gains, and cumulative weight gains
were observed in males receiving 250 mg/kg bw/day from Day 21 of the dosing period to Day 14 of t
he recovery period, from Day 7 to Day 35 of the dosing period, and from Day 14 to Day 42 of the dosi
ng period. Significant decreases in body weights, weight gains, and cumulative weight gains wereobserved in females receiving 250 mg/kg bw/day from Day 42 of the dosing period to Day 14 of therecovery period, from Day 21 to Day 35 of the dosing period, and from Day 35 to Day 42 of the dosing
period. No changes in body weights were observed in both sexes receiving 125 mg/kg bw/day and 6
2.5 mg/kg bw/day groups compared with the control groups during the dosing and recovery periods.
FOOD CONSUMPTION: Significant decreases in food consumption were observed in males receiving
250 mg/kg bw/day from Day 7 to Day 8 and Day 29 to Day 30 during the dosing period. Significant
decreases in food consumption were observed in females receiving 250 mg/kg bw/day of the sate13llite group from Day 1 to Day 2 and Day 29 to Day 30 during the dosing period. No changes in food consumption were observed in both sexes receiving 125 mg/kg bw/day and 62.5 mg/kg bw/day groups
compared with the control groups during the dosing and recovery periods.
URINALYSIS: Slight turbidity was observed in one male receiving 250 mg/kg bw/day during the dosing
period. Occult blood was observed in each one male in the 250 mg/kg bw/day group and the control
group during the dosing period. Red blood cells in sediments were observed in six males receiving
250 mg/kg bw/day and remained in one male during the recovery period.
HAEMATOLOGY: At the end of the dosing period, significant decreases in hematocrit value and In
creasing tendency in reticulocyte ratio were observed in males receiving 250 mg/kg bw/day. Nochanges were observed in female receiving 250 mg/kg bw/day. At the end of the recovery period, significant decreases in hemoglobin concentration, hematocrit value, MCV, and MCH were observed infemales receiving 250 mg/kg bw/day. No significant differences in males receiving 250 mg/kg bw/
day compared with the control group.
CLINICAL CHEMISTRY: At the end of the dosing period, significant decreases in albumin value wereobserved in males receiving 62.5 mg/kg bw/day and 250 mg/kg bw/day. A significant decrease increatinine value was observed in males receiving 250 mg/kg bw/day and remained after the recovery
period. No changes were observed in female receiving 250 mg/kg bw/day compare with the control
group after the dosing period. At the end of the recovery period, a significant decrease in a creatinine
value and a significant increase in triglyceride value were observed in females receiving 250 mg/kg
bw/day.
ORGAN WEIGHTS: At the end of the dosing, A significant decrease in body weight was observedin male receiving 250 mg/kg bw/day; therefore, significant increases in relative brain, heart, andadrenals weights were observed in this group. No changes in organ weight were observed in females.
At the end of the recovery period, significant decreases in absolute liver and kidneys weights andsignificant increases in relative brain, heart, kidneys, spleen, adrenals, testes, and epididymis wereobserved in males receiving 250 mg/kg bw/day. Significant decreases in absolute thymus weight andsignificant increases in relative brain, heart, and liver weights were observed in females receiving 250
mg/kg bw/day compared with the control group.
GROSS PATHOLOGY: Edematous and subinvolution in the lungs and enlargement and dark red colored in the kidneys were observed in the dead male. Edematous in the lungs and thymus, enlarges
and pale colored kidneys, pale colored and small sized spleen, and pale colored bone marrow offemur were observed in the dead female. At the end of the dosing period, abscess, yellow zone, andadhesion with scrotum in the epididymis were observed in one or two males receiving 250 mg/kg bw/
day. Enlargement and rough surface in the kidneys, small sized spleen, nodule and dark red zone in tai
l were observed in males receiving 250 mg/kg bw/day. Enlargement of the kidneys and white spot in
the liver were observed in males receiving 62.5 mg/kg bw/day. Small sized spleen and thymus, black
spot or recessed area in mucosa of the glandular stomach, and nodule and black spot of skin in theauricle were observed in females receiving 250 mg/kg bw/day. Cyst in the kidney, mass of aroundthe vulva in the subcutis, black spot of mucosa in the glandular stomach, and white spot in the liver
were observed in females receiving 125 mg/kg bw/day. At the end of the recovery period, abscessand adhesion with scrotum, nodule of scrotum in skin, and black spot of auricle in skin were observed in one male receiving 250 mg/kg bw/day. Alopecia of skin in clavicular area were observed in one female receiving 250 mg/kg bw/day.
HISTOPATHOLOGY: NON-NEOPLASTIC: Granuloma with multinucleated giant cells and inflammatory
cell infiltration was observed in subcutaneous tissues in lesion of the auricle, tail and scrotum. Necro
sis or edemas of the papilla of the kidneys were observed in males receiving 62.5 mg/kg bw/day andmore and females receiving 250 mg/kg bw/day groups. Deposit of pigment of proximal tubule in thekidneys was observed in both sexes receiving 62.5 mg/kg bw/day or more. Diffuse hypertrophy offascicular cells in the adrenal glands was observed in both sexes receiving 62.5 mg/kg bw/day or mo
re. Lesions in the kidneys or adrenal glands were observed in both sexes receiving 250 mg/kg bw/day
at the end of the recovery period.

14

Effect levels
Key result
false
Dose descriptor
LOAEL
Effect level
62.5

mg/kg bw/day (actual dose received)

Based ontest mat.
Sexmale/female
Basis for effect level
other: Effects of lesions in the deposit of pigment in proximal tubule in kidneys and diffuse hypertrophy in adrenals in males and females receiving 62.5 mg/kg bw/day or more groups.

Target system / organ toxicity
Key result
false
Critical effects observed
not specified

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/PDF638-16-4d.pdf

Applicant's summary and conclusion
Conclusions
Based on the effects of dosing on the kidney and adrenal gland, the LOAEL for the male and female rat repeated dose toxicity of 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione was determined to be 62.5 mg/kg
bw/day.
Executive summary
A combined repeated oral dose toxicity study with the reproduction/developmental toxicity screening
test was performed according to the OECD TG 422. Male and female rats (12 animals/sex/dose) wereadministered 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione at 0, 62.5, 125, and 250 mg/kg bw/day. Males were
dosed for 48 days, including a 14 day pre-mating period and subsequent mating period. Females were
dosed for up to 54 days, including 14 day pre-mating, mating, and gestation periods, and the time until
lactation day 4. Five out of 12 males at 0 and 250 mg/kg bw/day were treated as a recovery group.
In addition, 5 females/dose administered 0 and 250 mg/kg bw/day were dosed for 42 days without
15

mating and were treated as a recovery group. One male and one female died after 250 mg/kg bw/day
dosing. Clinical signs of toxicity included black areas on the pinna, dark purple coloration at the distal
end of the tail, reddish urine, induration of the scrotum, and nodules of the tail, pinna, and scrotum inthe 250 mg/kg bw/day group. Transient salivation was observed in males at 125 mg/kg bw/day and inboth sexes at 250 mg/kg bw/day. At 250 mg/kg bw/day, food consumption and body weight gain weredecreased in males and non-mating females. Red blood cells were observed in the urinary sedimentfrom 6 males in the 250 mg/kg bw/day group. In the blood, hematocrit and albumin were decreased inmales at 250 mg/kg bw/day. Gross pathological changes were observed in the tail, pinna and scrotum,
and the histopathological examination revealed granulation tissues with multinucleated giant cells andinflammatory cell infiltration in the subcutis of the tail, pinna, and scrotum. In the kidney, papilla necrosis
and edema were observed in males at doses of 62.5 mg/kg bw/day and higher, and in females at 250
mg/kg bw/day. Deposition of brown pigment in the basophilic tubule cortex was observed in both sexes
at doses of 62.5 mg/kg bw/day and higher. In the adrenal grand, diffuse hypertrophy of the fascicular
cells was observed in both sexes at doses of 62.5 mg/kg bw/day and higher. The histopathologicalchanges observed in the kidneys and adrenal grand did not resolve after the recovery period. Based on
the effects of dosing on the kidney and adrenal gland, the LOAEL for the male and female rat repeateddose toxicity of 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione was determined to be 62.5 mg/kg bw/day.

16

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

UUID: IUC5-6e3a7ad5-8227-4baa-bd43-d4991de751c6
Dossier UUID:
Author:Date: 2022-12-16T11:13:07.075+09:00
Remarks:

Administrative data
Endpoint
in vitro gene mutation study in bacteria Type of genotoxicity: gene mutation
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

Data source
Reference
Reverse Mutation Test of trithiocyanuric acid on Bacteria. / MHW (Ministry of Health and Welfare),Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)17

in vitro gene mutation study in bacteria
Deviations
no
Qualifieraccording to guideline
Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviationsyes
GLP compliance
yesType of assay
bacterial reverse mutation assay
in vitro gene mutation study in bacteria

Test material
Test material information
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

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
SD rat liver, induced by phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose-/+ S9 mix: 0, 313, 625, 1250, 2500, 5000 μg/plate (all test strains)
Vehicle / solvent
Dimethyl sulfoxide
Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes18True negative controlsnoPositive controls
yes
Positive control substanceother: -S9 mix: 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (TA 100, TA98 and WP2 uvrA/pKM101), s
odium azide (TA1535) and 9-aminoacridine hydrochloride (TA1537). +S9 mix: 2-aminoanthracene(all strains)
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
In a cytotoxicity test, growth inhibition was not observed at 50, 150, 500, 1500, 5000 μg/plate with orwithout S9 mix.
Evaluation criteria
A chemical was judged to be positive when the mean number of revertant colonies per plateincreased more than twice in comparison with that of the negative control and dose-responsibility andreproducibility was confirmed.
StatisticsnoResults and discussion
Test results
Key result
false
Species / strain
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
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

19

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
validRemarks on result
other: all strains/cell types tested Migrated from field 'Test system'.

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/PDF638-16-4e.pdf

Applicant's summary and conclusion
Conclusions
Interpretation of results (migrated information):negative
In a bacterial reverse mutation assay using Salmonella typhimurium TA100, TA1535, TA98, andTA1537, and Escherichia coli WP2uvrA (OECD TG 471), 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione was negative with or without metabolic activation.
Executive summary
No increase in revertant colonies was observed in the test with either the non-activation method (-S9)
or activation (+S9) method. Reverse mutation assays using microorganisms (Salmonella typhimurium,
Escherichia coli) were conducted to assess the potential of trithiocyanuric acid to induce genemutations. Trithiocyanuric acid did not induce gene mutations in the bacteria under the conditions of
this study. The positive control showed expected results.

20

ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.002

UUID: IUC5-0f48a174-5ea0-4c50-9a94-e9edcc53a764
Dossier UUID:
Author:Date: 2017-01-04T16:29:49.000+09:00
Remarks:

Administrative data
Endpoint
in vitro cytogenicity / chromosome aberration study in mammalian cells Type of genotoxicity:
chromosome aberration
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

Data source
Reference
In Vitro Chromosomal Aberration Test of Trithiocyanuric acid on Cultured Chinese Hamster Cells. /MHW (Ministry of Health and Welfare), Japan / study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)in vitro cytogenicity / chromosome aberration study in mammalian cells
21

Deviations
no
Qualifieraccording to guideline
Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviationsno
GLP compliance
yesType of assay
in vitro mammalian chromosome aberration test
chromosome aberration

Test material
Test material information
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

MethodSpecies / strain
Species / strain / cell typeother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
with and without
Metabolic activation system
SD rat liver, induced by phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose
-S9 mix (continuous treatment): 0.14, 0.21, 0.31, 0.47, 0.70 mg/mL-S9 mix (short-term treatment): 0.36, 0.53, 0.80, 1.2, 1.8 mg/mL+S9 mix (short-term treatment): 0.36, 0.53, 0.80, 1.2, 1.8 mg/mL
Vehicle / solvent- Vehicle(s)/solvent(s) used: DMSO
Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes
True negative controlsnoPositive controls
yes
22

Positive control substanceother: [continuous treatment -S9]: mitomycin C; [continuous treatment +S9]: Benzo[a]pyrene
Details on test system and experimental conditions
METHOD OF APPLICATION: Exposure duration: [continuous treatment]: 24 hrs [short-term treatment]:
No description
SPINDLE INHIBITOR: Colcemid
STAIN: Giemsa stain (3%)
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 200 cells / dose
DETERMINATION OF CYTOTOXICITY- Method: statistic method and biological evaluation by the relative total growth
Evaluation criteria
Increased uumber of cells with aberations were statisticaly examined.
Statistics
Fisher’s test (P < 0.01 by one-sided test) and Chochran-Armitage trend test (P < 0.01 by one-sided t
est)

Results and discussion
Test results
Key result
false
Species / strainother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
with
Genotoxicity
positive
Cytotoxicity / choice of top concentrations
cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not examined
Positive controls validity
valid
Key result
false
Species / strainother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
without
Genotoxicity
negative23

Cytotoxicity / choice of top concentrations
cytotoxicity
Vehicle controls validity
valid
Untreated negative controls validity
not valid
Positive controls validity
validRemarks on result
other: strain/cell type: Migrated from field 'Test system'.

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/PDF638-16-4f.pdf

Applicant's summary and conclusion
Conclusions
Interpretation of results (migrated information):
positive with metabolic activation
An in vitro chromosomal aberration test using CHL/IU cells (OECD TG 473) was positive with met
abolic activation.

24Genetic toxicity in vivo
ENDPOINT_STUDY_RECORD: Genetic toxicity in vivo.001

UUID: IUC5-b741db6c-1836-485b-9a8c-d9818db114bb
Dossier UUID:
Author:Date: 2022-12-14T15:47:04.148+09:00
Remarks:

Administrative data
Endpoint
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus Type of genotoxicity:
chromosome aberration
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: GLP guideline study

Data source
Reference
Micronucleus test of Trithiocyanuric acid on mouse / MHW (Ministry of Health and Welfare), Japan /study report
Data accessdata published

Materials and methods
Test guideline
Qualifieraccording to guideline
Guideline
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)25

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Deviations
no
Qualifieraccording to guideline
Guideline
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviationsno
GLP compliance
yesType of assay
micronucleus assay
chromosome aberration

Test material
Test material information
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
Specific details on test material used for the study
Name of test material (cited in study report): 2,4,6-trimercapto-S-triazine

Test animals
Speciesmouse
Strainother: Crlj:CD1(ICR)
Sexmale
Details on test animals or test system and environmental conditions
TEST ANIMALS- ICR, [CD1 (ICR), SPF- Source: Charles River Laboratories Japan, Inc.
- Age at study initiation: 9 weeks- Weight at study initiation: range findings, males: 33.9-38.1 g, females: 25.8-29.3 g: main study:
males: 32.4-39.2 g- Assigned to test groups randomly: yes- Fasting period before study: no- Housing: bracket type TPX resin cage, (143W×293D×148Hmm)- Diet: ad libitum- Water: ad libitum- Acclimation period: 9 days
ENVIRONMENTAL CONDITIONS- Temperature (°C): 21-25- Humidity (%): 40-75- Air changes (per hr): 10-15/h
26

- Photoperiod : 12 h dark/12 h light (light time: 8:00 AM to 8:00 PM)

Administration / exposure
Route of administration
oral: gavage
Vehicle- Vehicle(s)/solvent(s) used: 0.5%CMC- Concentration of test material in vehicle: 250, 500, 1000, and 2000 mg/mL- Amount of vehicle: 10 mL/kg bw
Details on exposure
RESULTS OF RANGE-FINDING STUDY- Dose range: 0, 250, 500, 1000, and 2000 mg/kg bw/day (24 h interval, twice)- Clinical signs of toxicity in test animals: Decreases in locomotor activity and piloerection were observed in one male receiving 2000 mg/kg bw/day and one female receiving 1000 mg/kg bw/day.
One male and one female mice died at 2000 mg/kg bw/day.
Duration of treatment / exposure
48 hFrequency of treatment
Twice, 24 h interval
Doses / concentrations
Remarks
Doses / Concentrations:
250, 500, 1000, and 2000 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose
3 males/doseControl animals
yes, concurrent vehicle
Positive control(s)Cyclophosphamide monohydrate (CP)- Route of dosing: oral gavage- Doses / concentrations: 50 mg/kg bw (single dose)

Examinations
Tissues and cell types examined
polychromatic erythrocytes from the femur bone marrow
Details of tissue and slide preparation
TREATMENT AND SAMPLING TIMES: Cells for specimen were collected 24 h after the last dosing.
DETAILS OF SLIDE PREPARATION: Cell suspensions were spread on a slide glass, dried, and fixed wi
th methanol for five min. Each specimen was stained with acridine orange.
27

METHOD OF ANALYSIS: fluorescence microscopy, blind method
Evaluation criteria
The test substance was determined to be positive if the micronucleated cells were statistically incr
eased in the dosing groups as compared with negative control group.
Statistics
Appearance frequency of micronuclei: Fisher’s test (one-sided test), Test was used to correct the Bo
nferroni in consideration of multiplicity. Significant level was 5 and 1% levels setting. Appearance
frequency of micronuclei was used trend test of Chochran-Armitage (one-sided test).
Polychromatic erythrocytes in erythrocytes: These rates were analyzed using Bartlett’s test forhomogeneity of distribution excluding positive control. Result of analysis, homogenous was observ
ed. Next, difference between negative control group and each treated groups wad analysed byDunnett’s multiple comparison test (one-sided test). Difference between negative and positive controls was analyses by F-test and Student’s t-test. These analyses method, Bartlett’s test and F-test
were setting 5% significant levels. Dunnett’s test and Student t-test were 5% and 1% levels setting.

Results and discussion
Test results
Key result
false
Sex
male
Genotoxicity
negative
Toxicity
yes
Vehicle controls validity
valid
Negative controls validity
not examined
Positive controls validity
valid
Additional information on results- Induction of micronuclei: appearance frequency of micronucleated cells (%MNPCE) for dose levelsof 0,250, 500, and 1000 mg/kg bw/day were 0.15%, 0.17%, 0.14%, and 0.13%, respectively.
- Ratio of PCE/NCE::ratio for dose levels, 0,250, 500, and 1000 mg/kg bw/day were 56.8%, 57.6%%,
59.1%, and 59.5%,, respectively.
- Body weight: not examined- Statistical evaluation: statistically significant increases were not observed.

Any other information on results incl. tables
Figures and Tables (in English) are available in the following full report of the study.
28

http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF638-16-4g.pdf

Applicant's summary and conclusion
Conclusions
Interpretation of results (migrated information): negative
The in vivo micronucleus study (OECD TG 474) was negative up to the maximum tolerated dose (1000mg/kg bw/day for 2 days) in mice.

29

Toxicity to reproduction
ENDPOINT_STUDY_RECORD: Toxicity to reproduction.001

UUID: IUC5-a60fe279-2ba7-4dea-9abd-4cbd6e5a6ee6
Dossier UUID:
Author:Date: 2022-12-16T11:14:13.699+09:00
Remarks:

Administrative data
Endpoint
screening for reproductive / developmental toxicity based on test type (migrated information)
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
Remarks
7.5.Repeated dose toxicity: oral: Repeated dose toxicity: oral.001

Data source
Reference
A combined repeated-dose/reproductive-developmental toxicity study of trithiocyanuric acid by oral a /MHW (Ministry of Health and Welfare), Japan / study report
Data accessdata published

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

Test material
Test material information
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Test animals
Speciesrat
Strainother: Crj: CD(SD), SPF
Sexmale/female

Administration / exposure
Route of administration
oral: gavage
Vehicleother: 0.5% sodium carboxymethyl cellulose
Details on exposure
VEHICLE- Justification for use and choice of vehicle: No data
- Amount of vehicle (if gavage): 5 ml/kg bw- Supplier: Maruishi pharmaceutical industry Co., Ltd.
- Lot/batch no. (if required): 4720
- Dosing volume: 5 mL/kg- Stability (test solutions): For 8 days
- Storage condition of test solution: Stored in a dark place with room temperature
Details on mating procedure- M/F ratio per cage:1:1- Length of cohabitation:up to two weeks- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as day 0 of pregnancy
Analytical verification of doses or concentrations
yes
31

Details on analytical verification of doses or concentrations
Test suspensions at each concentration (20 and 0.05 w/v%) of initial and final preparations wereanalyzed by the HPLC method at Hatano laboratory of Food and Drug Safety Center. Results show
ed that the concentration of the test article in each concentration was 94.3 to 101% of the nominal
concentration.
Duration of treatment / exposure(P) Males: 42 days including 14 days pre-mating, mating, and thereafter 14 periods (subsequent 28 days)(P) Females: Days including 14 days pre-mating, mating and gestation periods and the days until day
4 of lactation
Frequency of treatment
Daily: 7 times / week
Doses / concentrations
Remarks
Doses / Concentrations:
0 (vehicle), 62.5, 125 and 250 mg/kg bw/day
Basis:
actual ingested
No. of animals per sex per dose
12/sex/dose (main group), and 5 females/dose at 0 and 250 mg/kg bw/day (satellite group)
Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: A preliminary study (study No. R-04-006) was conducted to determine thedoses to be employed. Male and female rats were receiving 0, 250, 500, and 1000 mg/kg bw/day for
14 days. As a result, red colored urine was observed in more than half numbers of rats receiving 1000
mg/kg bw/day groups and all animals died in the 1000 mg/kg bw/day groups until Day 7. Black pathzones of auricle and dark purple colored tail top were observed in the 500 mg/kg bw/day groups from
Week 2, and moreover, one female sowing red colored urine, salivation, wasting and sedation were
moribund . Body weight gains were depressed in male rats receiving 500 mg/kg bw/day at Week 2,
and swelling of the spleen was observed at necropsy. No changes related to the test substance wereobserved in males receiving 250 mg/kg bw/day. From the results of the preliminary study, 500 mg/kg
bw/day seemed to be the maximum tolerance dose for 14 days. Therefore, the high dose was set at
250 mg/kg bw/day for the main study, and the middle and low doses were set at 125 and 62.5 mg/kg
bw/day using common ratio 2..

Examinations
Parental animals: Observations and examinations
CAGE SIDE OBSERVATIONS: Yes- Time schedule:
Males and females: once before the start of dosing, two times/day during the dosing period, and once
during the recovery period
DETAILED CLINICAL OBSERVATIONS: Yes- Time schedule:
Male animals: the end of acclimation period, Day 7, 14, 21, 28, 35 and 42 during the dosing and Day
7 and 14 during the recovery period. Female animals: the end of acclimation period, Day 7, 14, 21,
28, 35 and 42 during the dosing, Day 0 at delivery period and Day 0 from 4 at delivery animals, andDay 7 and 14 of satellite groups during the recovery period. Test items are following: Body position, lo
comotor activity, vocalization, tremor, convulsion by cage side observation, ease of removing rat from32

cage, reactivity to being handled, heart rate, body temperature, fur, skin color, visible mucosa color,
lacrimation, exophthalmos ophthalmocele, pupillary, and salivation by terminology for removing rat
from cage, and body position, walking, grooming, phonation, straub tail, gait, stereotyped, abnormal
behavior, tremor, convulsion, piloerection, and ophthalmorrhexis by observation of behavior on theworking table.
BODY WEIGHT: Yes
Males in the main and females in satellite groups were weighed on Day 1, 7, 14, 21, 28, 35 and 42 of
dosing, and Day 1, 7, and 15 (necropsy day) at the end of the recovery period.
Females were weighed on Day 1, 7, 14, and 21 until successful copulation. Copulated females were
weighed on Day 0, 7, 14 and 20 of gestation, days 0 and 4 of lactation, and the necropsy day.
FOOD CONSUMPTION : Yes- Food consumption (g/day/rat) for each animal determined from the difference of the of the previous
day's feeding amount: Yes
Measurement of food consumption was conducted on all animals at the following frequencies:
Males and females in the satellite groups: on Day 1-2, 7-8, 14-15, 29-30, 35-36, and 41-42 during thedosing period and Day 6-7 and 13-14 during the recovery period. Females in the main groups: on Day
1-2, 7-8, and 14-15 (before mating), on Day 0-1, 7-8, 14-15, and 20-21 (the gestation period). On Day
3-4 (the lactation period).
FOOD INTAKE: No
COMPOUND INTAKE: No
FOOD EFFICIENCY: No
WATER CONSUMPTION: No
Oestrous cyclicity (parental animals)
Vaginal smears were collected from all females in the main groups and microscopically examined
every day from the day after the start of dosing until the day copulation was confirmed. Mean estrous
cycle (day) and abnormal estrous cycle animals were examined by dams.
Sperm parameters (parental animals)
testis weight, epididymis weight
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.
Postmortem examinations (parental animals)
METHOD OF SACRIFICED: All animals were sacrificed by exsanguination under pentobarbital sodium
anesthesia, intraperitoneally.
SACRIFICE: Male animals: On Day 42, Maternal animals: on Day 4 of lactation, and Male recovery andfemale satellite animals: on Day 56.
GROSS PATHOLOGY AND ORGAN WEIGHTS：Yes Brain, heart, liver, kidneys, adrenals, thymus, sp
leen, testes, and epididymis.
HISTOPATHOLOGY: Yes Brain, puitality, thymus, lymph nodes (including mesenteric and mandibularlymph nodes), trachea, lung (including bronchus), stomach, intestinal tract (duodenum, jejunum,
ileum, cecum, colon, rectum), thyroids, heart, liver, spleen, kidneys, adrenals, urinary bladder, testes,
epididymis, seminal vesicles (including the coagulating gland), prostate (ventral lobe), ovaries, uteru
s, vagina, bone marrow (one side femur), sciatic nerve (one side femur), spinal cord, and gross abnormalities site.
Postmortem examinations (offspring)
SACRIFICE: F1 pups were euthanized on PND 4 by exsanguination pentobarbital sodium anesthesia,
intraperitoneally.
GROSS NECROPSY: Yes

33

Statistics
Changes in estrous cyclicity and conception rate were analyzed by Fisher’s test. Graded pathological
data was analyzed by Mann-Whitney’s U test (significance level = 0.05) and pathological data with number of positive and negative animals was analyzed by one-sided Fisher’s test.
Other data, obtained values in each animal or mean of a litter was one data, and these data were compared among the satellite groups and other among the groups. These data were analyzed using Ftest for homogeneity of distribution. The Student’s t-test and the Aspin-Welch’s t-test were conducted
for homogenous and non-homogenous distribution, respectively to compare the control and individual treatment groups. Three or more groups setting, these data were analyzed using Bartlett’s testfor homogeneity of distribution. The Dunnett’s multiple comparison test after the ANOVA and the Du
nnett’s-type mean rank sum test after Kruskal-Wallis’s H test were conducted for homogenous andnon-homogenous distribution, respectively to compare the control and individual treatment groups.
Significance level was set at 0.05 compared with the control group and among the groups.
Reproductive indices
1) Each parameter was determined by the following equations:
Mean estrus cycle, incidence of females with irregular estrus cycle, mating periods,
Copulation index (%) = (No. of copulated animals/No. of co-housed animals) × 100
Fertility index (%) = (No. of pregnant females/No. of copulated females) × 100
Gestation length, number of corpora lutea, number of implantation sites, total number of offspring,
Implantation index (%) = (No. of implantation sites/No. of corpora lutea) × 100
Delivery index (%) = (No. of females delivered liveborn pups/No. of pregnant females) × 100
Gestation index (%) = (No. of pregnant animals delivered live offspring/number of pregnant animals) ×100
Offspring viability indicesTotal number of offspring at birth, number of live offspring at birth,
Number of live pups on day 0 of lactationBirth index (%) = (Number of live pups on day 0/Number of
implantation sites) ×100
Viability index = (Number of live pups on day 4 after birth/Number of live pups born) ×100
External examination of offspring, necropsy finding
Pups weight on day 0 of lactation
Sex ratio on day 0 of lactation
Number of live pups on day 4 of lactation
Pups weight on day 4 of lactation
Sex ratio on day 4 of lactation

Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
Clinical signs
effects observed, treatment-related
Body weight and weight changes
effects observed, treatment-related
Food consumption and compound intake (if feeding study)
effects observed, treatment-related
Organ weight findings including organ / body weight ratios
no effects observed34

Gross pathological findings
effects observed, treatment-related
Histopathological findings: non-neoplastic
effects observed, treatment-related

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

Details on results (P0)1) Estrous Cycle
A few changes in estrous cycles were observed in females receiving 125 mg/kg bw/day and 250 mg/
kg bw/day, however these frequencies were not significantly different compared with the control group.
2) Results of Mating
Mating was successful in all the animals, but Infertility was observed in three females receiving 250
mg/kg bw/day.
3) Delivery Data and Delivery
There were no significant differences in the gestation length, number of implantation sites, impl
antation index, and delivery index between the control group and any treatment groups. Significant decreases in number of corpora lutea were observed in females receiving 250 mg/kg bw/day
GROSS PATHOLOGY
See 7.5.1 Repeated dose toxicity: oral
HISTOPATHOLOGY
See 7.5.1 Repeated dose toxicity: oral

Effect levels (P0)
Key result
false
Dose descriptor
NOAEL
Effect level125

mg/kg bw/day (actual dose received)

Based ontest mat.
Sex
femaleBasis for effect level
other: Infertility was observed in 3 females at 250 mg/kg bw/day. The number of corpora lutea dec
reased in rats given 250 mg/kg bw/day

Results: F1 generation
35

General toxicity (F1)Clinical signs
no effects observed
Mortality / viability
no mortality observed
Body weight and weight changes
no effects observed

Details on results (F1)no effects

Overall reproductive toxicity
Key result
false
Reproductive effects observed
not specified

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/PDF638-16-4d.pdf

Applicant's summary and conclusion
Conclusions
The NOAEL for the rat reproductive/developmental toxicity of 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione
was determined to be 125 mg/kg bw/day based on infertility and a decrease in corpora lutea.
Executive summary
In the combined repeated oral dose toxicity study with the reproduction/developmental toxicity
screening test (OECD TG 422), infertility was observed in 3 females at 250 mg/kg bw/day. The numberof corpora lutea decreased in rats given 250 mg/kg bw/day. No effects were observed in any pups. The
NOAEL for the rat reproductive/developmental toxicity of 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione wasdetermined to be 125 mg/kg bw/day based on infertility and a decrease in corpora lutea.

36

DOMAIN
Substance
SUBSTANCE: 1,3,5-triazine-2,4,6(1H,3H,5H)-trithione
UUID: IUC5-80e43b49-60cb-49c6-87bc-4abf9b428515
Dossier UUID:
Author:Date: 2022-12-16T11:14:34.696+09:00
Remarks:
Substance name
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione
Legal entity
National Institute of Health Sciences / Kawasaki / Japan

Identification of substance
Reference substance
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
EC number

EC name

211-322-8

EC Inventory

CAS number

CAS name638-16-4
IUPAC name
1,3,5-triazinane-2,4,6-trithioneRole in the supply chain
Manufacturer
false
Importer
false
Only representativefalse
Downstream user
false

37

References
Reference Substances
REFERENCE_SUBSTANCE: 1,3,5triazine-2,4,6(1H,3H,5H)-trithione
UUID: ECB5-c72a77ea-657c-4fbf-a95d-f61240e1c79d
Dossier UUID:
Author:Date: 2007-05-10T18:00:00.000+09:00
Remarks:
Reference substance name
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione
IUPAC name
1,3,5-triazinane-2,4,6-trithione

Inventory
Inventory number
Inventory name
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione
Inventory
EC Inventory
Inventory number
211-322-8
CAS number
638-16-4
Molecular formula
C3H3N3S3
Description
CAS number
638-16-4

Synonyms
Synonyms
Identity
1,3,5-triazine-2,4,6(1h,3h,5h)-trithione
Identity
1,3,5-Triazine-2,4,6(1H,3H,5H)-trithione

Molecular and structural information
38

Molecular formula
C3H3N3S3
Molecular weight
177.271
SMILES notation
S=C1NC(=S)NC(=S)N1
InChl
InChI=1/C3H3N3S3/c7-1-4-2(8)6-3(9)5-1/h(H3,4,5,6,7,8,9)
Structural formula

Related substancesGroup / category information
USEPA Category: Substituted Triazines;Thiols

39

Test Materials
TEST_MATERIAL_INFORMATION: 1,3,5-triazinane-2,4,6trithione / 638-16-4 / 211-322-8
UUID: 1cc62be1-b4ac-3ddf-9266-ccd0243f7850
Dossier UUID:
Author:Date: 2017-01-04T16:29:25.000+09:00
Remarks:
Name
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Composition
Composition
Type
Constituent
Reference substance
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
EC number

EC name

211-322-8

EC Inventory

CAS number

CAS name638-16-4
IUPAC name
1,3,5-triazinane-2,4,6-trithione

Other characteristics
Details on test material- Name of test material (as cited in study report): Trithiocyanuric acid
- CAS No.: 638-16-4- Molecular formula: C3H3N3S3- Molecular weight: 177.27- Purity: 99.8% (HPLC)- Impurities: 0.13% sulfur- Physical state: Slightly pale yellow powder- Melting point: >300°C- Boiling point, specific gravity, partition coefficient, vapor pressure: No data
- Solubility: Sparingly soluble in water, slightly soluble in methanol, acetone, and dioxane, and soluble in cellosolve and THF- Stability: The stable to normal handling- Supplier: Kawaguchi Chemical Industry Co., Ltd.
- Lot No.: 407518- Storage condition until use: Room temperature

40

TEST_MATERIAL_INFORMATION: 1,3,5-triazinane-2,4,6trithione / 638-16-4 / 211-322-8
UUID: 7f161f9c-ae98-3924-aa62-73524d965804
Dossier UUID:
Author:Date: 2017-01-04T16:30:03.000+09:00
Remarks:
Name
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Composition
Composition
Type
Constituent
Reference substance
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
EC number

EC name

211-322-8

EC Inventory

CAS number

CAS name638-16-4
IUPAC name
1,3,5-triazinane-2,4,6-trithione

Other characteristics
Details on test material- Name of test material (as cited in study report): Trithiocyanuric acid
- CAS No.: 638-16-4- Molecular formula: C3H3N3S3- Molecular weight: 177.27- Purity: 99.8% (HPLC)- Impurities: 0.13% sulfur- Physical state: Slightly pale yellow powder- Melting point: >300°C- Boiling point, specific gravity, partition coefficient, vapor pressure: No data
- Solubility: Sparingly soluble in water, slightly soluble in methanol, acetone, and dioxane, and solublein cellosolve and THF- Stability: The stable to normal handling- Supplier: Kawaguchi Chemical Industry Co., Ltd.
- Lot No.: 407518- Storage condition until use: Room temperature

41

TEST_MATERIAL_INFORMATION: 1,3,5-triazinane-2,4,6trithione / 638-16-4 / 211-322-8
UUID: 4ddf3c8e-6843-3714-9bc2-20503be39c0b
Dossier UUID:
Author:Date: 2017-01-04T16:29:34.000+09:00
Remarks:
Name
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Composition
Composition
Type
Constituent
Reference substance
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
EC number

EC name

211-322-8

EC Inventory

CAS number

CAS name638-16-4
IUPAC name
1,3,5-triazinane-2,4,6-trithione

Other characteristics
Details on test material- Name of test material (as cited in study report): Trithiocyanuric acid
- CAS No.: 638-16-4- Molecular formula: C3H3N3S3- Molecular weight: 177.27- Purity: 99.8% (HPLC)- Impurities: 0.13% sulfur- Physical state: Slightly pale yellow powder- Melting point: ＞300°C- Boiling point, specific gravity, partition coefficient, vapor pressure: No data
- Solubility: Sparingly soluble in water, slightly soluble in methanol, acetone, and dioxane, and solublein cellosolve and THF- Stability: The stable to normal handling- Supplier: Kawaguchi Chemical Industry Co., Ltd.
- Lot No.: 407518- Storage condition until use: Room temperature

42

TEST_MATERIAL_INFORMATION: 1,3,5-triazinane-2,4,6trithione / 638-16-4 / 211-322-8
UUID: aa3065f1-b8b3-3f79-9a41-c5d6004bfc5a
Dossier UUID:
Author:Date: 2017-01-04T16:29:49.000+09:00
Remarks:
Name
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Composition
Composition
Type
Constituent
Reference substance
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
EC number

EC name

211-322-8

EC Inventory

CAS number

CAS name638-16-4
IUPAC name
1,3,5-triazinane-2,4,6-trithione

Other characteristics
Details on test material- Name of test material (as cited in study report): Trithiocyanuric acid
- CAS No.: 638-16-4- Molecular formula: C3H3N3S3- Molecular weight: 177.27- Purity: 99.8% (HPLC)- Impurities: 0.13% sulfur- Physical state: Slightly pale yellow powder- Melting point:> 300°C- Boiling point, specific gravity, partition coefficient, vapor pressure: No data
- Solubility: Sparingly soluble in water, slightly soluble in methanol, acetone, and dioxane, and soluble in cellosolve and THF- Stability: The stable to normal handling- Supplier: Kawaguchi Chemical Industry Co., Ltd.
- Lot No.: 407518- Storage condition until use: Room temperature

43

TEST_MATERIAL_INFORMATION: 1,3,5-triazinane-2,4,6trithione / 638-16-4 / 211-322-8
UUID: 112cb490-c985-347f-a67a-75b77768ba53
Dossier UUID:
Author:Date: 2022-12-14T16:01:14.616+09:00
Remarks:
Name
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Composition
Composition
Reference substance
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
EC number

EC name

211-322-8

EC Inventory

CAS number

CAS name638-16-4
IUPAC name
1,3,5-triazinane-2,4,6-trithione

Other characteristics
Details on test material- Name of test material (as cited in study report): Trithiocyanuric acid
- CAS No.: 638-16-4- Molecular formula: C3H3N3S3- Molecular weight: 177.27- Purity: 99.8% (HPLC)- Impurities: 0.13% sulfur- Physical state: Slightly pale yellow powder- Melting point:> 300°C- Boiling point, specific gravity, partition coefficient, vapor pressure: No data
- Solubility: Sparingly soluble in water, slightly soluble in methanol, acetone, and dioxane, and soluble in cellosolve and THF- Stability: The stable to normal handling- Supplier: Kawaguchi Chemical Industry Co., Ltd.
- Lot No.: 407518- Storage condition until use: Room temperature

44

TEST_MATERIAL_INFORMATION: 1,3,5-triazinane-2,4,6trithione / 638-16-4 / 211-322-8
UUID: c6be500f-a596-303a-a70d-da93de6e578a
Dossier UUID:
Author:Date: 2017-01-04T16:30:18.000+09:00
Remarks:
Name
1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8

Composition
Composition
Type
Constituent
Reference substance
1,3,5-triazine-2,4,6(1H,3H,5H)-trithione / 1,3,5-triazinane-2,4,6-trithione / 638-16-4 / 211-322-8
EC number

EC name

211-322-8

EC Inventory

CAS number

CAS name638-16-4
IUPAC name
1,3,5-triazinane-2,4,6-trithione

Other characteristics
Details on test material- Name of test material (as cited in study report): Trithiocyanuric acid
- CAS No.: 638-16-4- Molecular formula: C3H3N3S3- Molecular weight: 177.27- Purity: 99.2%- Physical state: Slightly pale yellow powder- Supplier: Wako Pure Chemical Industries, Ltd.
- Lot No.: KWM0501- Storage condition until use: Room temperature (17.0-23.2°C)

45

Literatures
LITERATURE: A combined repeated-dose/reproductivedevelopmental toxicity study of trithiocyanuric acid byoral administration in rats.
UUID: 6db66ae3-4239-3ec3-b05c-5b4ebfe32d4e
Dossier UUID:
Author:Date: 2017-01-04T16:30:03.000+09:00
Remarks:

General information
Reference Typestudy report
Title
A combined repeated-dose/reproductive-developmental toxicity study of trithiocyanuric acid by oral administration in rats.
AuthorMHW (Ministry of Health and Welfare), Japan
Year
2007
Bibliographic source
Japan Existing Chemical Data Base (JECDB)
Testing facility
Food and Drug Safety Center
Study number
R-04-007

46

LITERATURE: In Vitro Chromosomal Aberration Test ofTrithiocyanuric acid on Cultured Chinese Hamster Cells.
UUID: 01544345-7007-3fb0-a7ca-9655250d7e7c
Dossier UUID:
Author:Date: 2017-01-04T16:29:49.000+09:00
Remarks:

General information
Reference Typestudy report
Title
In Vitro Chromosomal Aberration Test of Trithiocyanuric acid on Cultured Chinese Hamster Cells.
AuthorMHW (Ministry of Health and Welfare), Japan
Year
2007
Bibliographic source
Japan Existing Chemical Data Base (JECDB)
Testing facility
Food and Drug Safety Center
Study number
G-04-065

47

LITERATURE: Micronucleus test of Trithiocyanuric acidon mouse
UUID: f531482c-693f-3b5a-b0fd-6cf9ac151c22
Dossier UUID:
Author:Date: 2017-01-04T16:30:18.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Micronucleus test of Trithiocyanuric acid on mouse
AuthorMHW (Ministry of Health and Welfare), Japan
Year
2010
Bibliographic source
Japan Existing Chemical Data Base (JECDB)
Testing facility
Food and Drug Safety Center
Study number
G-09-021

48

LITERATURE: Reverse Mutation Test of trithiocyanuric
acid on Bacteria.
UUID: 50307a75-a19c-3293-8805-666e4e4e1f9a
Dossier UUID:
Author:Date: 2017-01-04T16:29:42.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Reverse Mutation Test of trithiocyanuric acid on Bacteria.
AuthorMHW (Ministry of Health and Welfare), Japan
Year
2007
Bibliographic source
Japan Existing Chemical Data Base (JECDB)
Testing facility
Food and Drug Safety Center
Study number
M-04-075

49

LITERATURE: Single Dose Oral Toxicity Test ofTrithiocyanuric Acid in RatsUUID: 9c90037a-d506-35bc-ac16-4d03353e2a2d
Dossier UUID:
Author:Date: 2017-01-04T16:29:25.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Single Dose Oral Toxicity Test of Trithiocyanuric Acid in Rats
AuthorMHW (Ministry of Health and Welfare), Japan
Year
2007
Bibliographic source
Japan Existing Chemical Data Base (JECDB)
Testing facility
Food and Drug Safety Center
Study number
A-04-07450

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 systemIUCLID451

ID
16558402024DIV750

52