Name: COMPLETE / SUBSTANCE : perfluoro-1,2-dimethylcyclohexane / 1,1,2,2,3,3,4,4,5,6-

decafluoro-5,6-bis(trifluoromethyl)cyclohexane / 306-98-9 Fri, 16 Dec 2022, 16:31:01+0900 /Legal entity owner: National Institute of Health Sciences
Printing date: 2022-12-16T16:31:01.918+09:00

iTable of Contents
0/0 ............................................................................................................................. 1
National Institute of Health Science ..................................................................... 2
perfluoro-1,2-dimethylcyclohexane ........................................................................ 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 ......................................................... 12
Genetic toxicity in vitro.001 ................................................ 12
Genetic toxicity in vitro.002 ................................................ 1773 Toxicity to reproduction ........................................................ 20
Toxicity to reproduction.001 ............................................... 20
DOMAIN .................................................................................................... 29
Substance ......................................................................................... 29
Substance ................................................................................ 29
References ................................................................................................................ 30
Reference Substances ...................................................................................... 30
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexane .................. 30
Literatures ....................................................................................................... 32
A combined repeated dose/reproductive developmental toxicity study ofperfluoro-1,2-dimethylcyclohexane cyclohexane by oral administration in
rats. .......................................................................................................... 32
In Vitro Chromosomal Aberration Test of on perfluoro-1,2dimethylcyclohexane Cultured Chinese Hamster Cells ................................. 33
Reverse Mutation Test of perfluoro-1,2-dimethylcyclohexane ....................... 34
Legal Entities ................................................................................................... 35
National Institute of Health Sciences .......................................................... 35ii

DOSSIER:UUID: 0
Dossier UUID:
Author:Date: 2022-12-16T16:31:01.802+09:00
Remarks:

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

Dossier subject
Dossier subject
perfluoro-1,2-dimethylcyclohexane / 1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexane /306-98-9
Public nameSubmitting legal entity
National Institute of Health Science
Dossier creation date/time
Fri, 16 Dec 2022, 16:31:01+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perfluoro-1,2-dimethylcyclohexane
CORE
General information
Assessment approach (assessment entities)
FIXED_RECORD: Assessment approach

UUID: a165ff3b-4bd5-3920-8cc2-87bd649de09e
Dossier UUID:
Author:Date: 2019-03-27T09:47:42.000+09:00
Remarks:3

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

UUID: 8978d9e2-7ec0-4817-be88-3d30ea58bcb8
Dossier UUID:
Author:Date: 2022-12-16T16:28:05.305+09:00
Remarks:

Administrative data
Endpoint
repeated dose toxicity: oral, otherType of information
experimental study
Adequacy of study
key study
Robust study summary
false
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1
Cross-reference
Reason / purpose for cross-reference
reference to same study
Related information
OECD / Toxicity to reproduction / Toxicity to reproduction.001 / perfluoro-1,2-dimethylcyclohexane /
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexa

Data source
Reference
A combined repeated dose/reproductive developmental toxicity study
dimethylcyclohex / Ministry of Health, Labor and Welfare, Japan / study report
4ofperfluoro-1,2-

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)
Deviationsno
GLP compliance
yes
Limit test
no

Test material
Specific details on test material used for the study- Name of test material (as cited in study report): Perfluoro-1, 2-dimethylcyclohexane- Analytical purity: 97%- Storage condition of test material: at a room temperature (15-23°C), protected from light- Stability under test conditions: The stability of test material was identified by analysis of the remai
nder.

Test animals
Speciesratcommon rodent species
Strainother: Crl:CD(SD)Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source: Charles River Japan, Inc., Atsugi Breeding Center.
- Age at study initiation: 11 weeks old- Weight at study initiation: Male: 397 g (364 -442 g), Female: 244 g (226-268 g)- Housing: Animals were individually housed in bracket-type metallic wire-mesh cages (254W × 350D
× 170H mm), from gestation day 17 to lactation day 4, Dams were bred individually or with individual
littermates in plastic cages (340W x 400D x 185H mm) and bedding.
- Diet: Solid feed (NMF: Oriental Yeast Co., ltd.) was given ad libitum.
- Water: Tap water was given ad libitum.
- Acclimation period: 20 days
5

ENVIRONMENTAL CONDITIONS- Temperature (°C): 23±3 (actual temperature: 21-28°C)- Humidity (%): 50±20% (actual humidity: 33-47%)- Air changes (per hr): 10-15- Photoperiod (hrs dark / hrs light): 12 hr dark/12 hr light (light: 7:00~19:00)

Administration / exposure
Route of administration
oral: gavage
Vehicleother: 0.5 w/v% CMC-Na solution mixed containing 0.1 v/v% Tween 80
Details on oral exposure- Amount of vehicle (if gavage): 5 mL/kg- Dosing volume: 5 mL/kg
Analytical verification of doses or concentrations
yes
Duration of treatment / exposure(P) Males: 42 days including 14 days pre-mating(P)Females: 41-45 days including 14 days pre-mating, mating and gestation periods and the days until
day 4 of lactation
Female (no mating, satellite group): 42 days
Frequency of treatment
Once/day, 7 days/week
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
Main group:12 animals/sex/dose
Satellite (Recovery) group: 5 males/dose and 5 females/dose (0 and 1000 mg/kg bw/day)6

Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: Based on the results of a 14-day preliminary study, the highest dose of1000 mg/kg bw/day was selected as an expected obvious toxic dose, and the lowest dose of 100 mg/
kg bw/day was selected as an expected no toxic dose. The middle dose levels of 300 mg/kg bw/day
were selected.
- Rationale for animal assignment (if not random): Body weight-balanced randomization[14-day preliminary study]
A 14-day repeated dose oral toxicity test (Crl:CD(SD) rats, doses: 0, 100, 300 or 1000 mg/kg bw/day). No death was observed even at 1000 mg/kg bw/day, and no effects were observed in any groups
except for a slightly decreased in MCH in males at 300 and 1000 mg/kg bw/day.

Examinations
Observations and examinations performed and frequency
CAGE SIDE OBSERVATIONS: Yes- Time schedule: 3 times/day (before administration, immediately after administration and 2 hours
after administration) during the administration period. Once a day during the recovery period.
DETAILED CLINICAL OBSERVATIONS: Yes- Time schedule:
Male main group: once before the start of administration, once every weekly during the administration
and recovery periods.
Female main group: once before the start of administration, days 1, 7, 14 and 20 of gestation, and day
4 of lactation.
Male and female recovery group: once before the start of administration, once every weekly during theadministration and recovery periods.
BODY WEIGHT: Yes- Time schedule for examinations:
Males in the main groups were weighed on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 ofadministration and on the day of necropsy, and 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 in addition to the meas
urement days for males in the main groups.
Females in the main groups were weighed on days 1, 4, 8, 11 and 15 of administration (uncopulated
animals were weighed on day 18 of administration as well), days 0, 4, 7, 11, 14, 17 and 20 of gestation, days 0 and 4 of lactation and the day of necropsy.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):- Food consumption: Yes
Measurement of food consumption was conducted on all animals at the following frequencies:
males in the main groups on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration; males and fem
ales in the recovery groups on days 1, 4, 8, 11 and 14 of recovery in addition to the measurement da
ys for males in the main groups; and females in the main groups on days 1, 4, 8, 11 and 15 of admini
stration, days 1, 4, 7, 11, 14, 17 and 20 of gestation and days 2 and 4 of lactation.
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes- Time schedule for collection of blood: At the end of administration period, or at the end of recovery
period in both sexes- Anaesthetic used for blood collection: ether
7

- Animals fasted: Yes- 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 percentage, platelet count, white blood cell count, differential white blood cell count, absolute number of each
white blood cell, prothrombin time, activated partial thromboplastin time, fibrinogen.
CLINICAL CHEMISTRY: Yes- Time schedule for collection of blood: At the end of administration period, or at the end of recovery p
eriod in both sexes- Animals fasted: Yes- How many animals: 5 animals/sex/group- Parameters checked: ALP, total cholesterol, triglyceride, phospholipids, total bilirubin, glucose, bloodurea nitrogen, creatinine, sodium, potassium, chloride, calcium, inorganic phosphorus, total protein,
albumin, A/G ratio, AST (GOT), ALT (GPT), LDH, γ-GTP
URINALYSIS OF MALES: Yes- Time schedule for collection of urine: final week of administration (days 39 to 40 of administration)and in the final week of recovery (days 11 to 12 of recovery)- Metabolism cages used for collection of urine: Yes
A urine collector to collect four-hour urine samples under fasting but ad libitum drinking conditions,followed by collection of 20-hour urine samples under ad libitum feeding and drinking conditions.
- How many animals: 5 animals/group- Parameters checked: pH, protein, ketones, glucose, occult blood, bilirubin, urobilinogen, color, sedim
ent, urine volume (4-hour volume), osmotic pressure, urine volume (20-hour volume), water intake (24hour volume)
NEUROBEHAVIOURAL EXAMINATION: Yes- Time schedule for examinations: Manipulative test and measurements of grip strength and motor
activity were conducted on 5 animals per group with the following frequencies: males in the maingroups were examined in the final week of administration (day 37 of administration), females in the main groups on day 4 of lactation (day 41 to day 44 of administration) after necropsy of F1 pups,
and males and females in the recovery groups in the final week of administration (day 37 of administ
ration) and in the final week of recovery (day 9 of recovery).
- Battery of functions tested:
1) Open field observation. Arousal, gait, posture, tremor, convulsion, rearing count, defecation(defecation count, urination), stereotypy (grooming, circling, etc.), abnormal behavior (self-biting,backward walking, etc.)
2) Manipulative Test. Auditory response, approach response, touch response, tail pinch response,
pupillary reflex, aerial righting reflex, landing foot splay
3) Measurement of Grip Strength. Following manipulative test, grip strength of forelimb and hind limb
was measured by CPU gauge MODEL-9502A (AIKOH Engineering Co., Ltd.).
4) Measurement of Motor Activity. Following measurement of grip strength, motor activity wasmeasured by a motor activity sensor for experimental animals NS-AS01 (Neuro Science, Inc). The
measurement was conducted for 1 hour, and measured values at 10-minute intervals and from 0 to
60 minutes were collected.
Sacrifice and pathology
GROSS PATHOLOGY: Yes
ORGAN WEIBHT: Yes [brain, thyroids including parathyroids), adrenal gland, thymus, spleen, heart,
liver, kidney, testis, epididymis]
HISTOPATHOLOGY: Yes, [cerebrum, cerebellum, pituitary, spinal cord (thoracic), sciatic nerve, thyroid,parathyroid, adrenal glands, thymus, spleen, submandibular lymph nodes, mesenteric lymph nodes, heart, lung (including bronchial), stomach, duodenum, jejunum, ileum, cecum, colon, rectum, liv
er, kidney, bladder, testis, epididymis, ovary, uterus, prostate, seminal vesicles, sternum and femur(including bone marrows), macroscopic lesions, and parts for identification (auricles)]8Statistics
The data were analyzed for homogeneity of variance by the Bartlett test. If variances were
homogeneous, data was analyzed by the Dunnett test, whereas heterogeneous data was analyzed by
the Dunnett type mean rank test (p<0.05, two-sided).
In the recovery test, these values of two groups were analyzed by F test. If variances were homogeneo
us, data was analyzed by the Student t-test, whereas heterogeneous data was analyzed by the AspinWelch t-test (p<0.05, two-sided).

Results and discussion
Results of examinations
Clinical signs
no effects observed
Mortality
no mortality observed
Body weight and weight changes
no effects observed
Food consumption and compound intake (if feeding study)no effects observed
Food efficiency
not examined
Water consumption and compound intake (if drinking water study)
not examined
Ophthalmological findings
not examined
Haematological findings
effects observed, treatment-related
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
Histopathological findings: neoplastic
not examined
Details on results
CLINICAL SIGNS AND MORTALITY:
Mortality: There was no death.
Clinical signs: There were no effects related to the test substance.
9

DETAILED CLINICAL OBSERVATIONS, MANIPULATIVE TEST, GRIP STRENGTH TEST AND LOC
OMOTOR ACTIVITY MEASUREMENT: There were no changes related to the test substance.
BODY WEIGHT:
There were no changes related to the test substance in any groups at the dosing and recovery
periods.
FOOD CONSUMPTION:
There were no changes related to the test substance in any groups at the dosing and recovery
periods.
URINALYSIS:There were no changes related to the test substance in any groups at the dosing and recovery pe
riods.
HAEMATOLOGY:[At the end of dosing period]: At 1000 mg/kg bw/day, decrease in hemoglobin and hematocrit were ob
served in males.
[At the end of recovery period]: There were no changes related to the test substance in any groups.
CLINICAL CHEMISTRY:
There were no changes related to the test substance in any groups at the dosing and recovery periods.
ORGAN WEIGHTS:
There were no changes related to the test substance in any groups at the dosing and recovery
periods.
GROSS PATHOLOGY:
There were no changes related to the test substance in any groups at the dosing and recovery periods.
HISTOPATHOLOGY: NON-NEOPLASTIC:[At the end of dosing period]: At 1000 mg/kg bw/day, minimal hypertrophy of centrilobular hepat
ocytes were observed in males.
[At the end of recovery period]: There were no changes related to the test substance in any groups.

Effect levels
Key result
true
Dose descriptor
NOAEL
Effect level300

mg/kg bw/day (actual dose received)

Based ontest mat.
Sexmale/female
10Basis for effect level
haematology
At 1000 mg/kg bw/day, decrease in hemoglobin and hematocrit were observed in males.
histopathology: non-neoplastic
At 1000 mg/kg bw/day, minimal hypertrophy of centrilobular hepatocytes were observed in males.

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/PDF595-90-4d.pdf

Applicant's summary and conclusion
Executive summary
A combined repeated-dose toxicity study with a reproduction/developmental toxicity screening test
(OECD TG422) was conducted to specify the general toxic effects of repeated administrationof perfluoro-1,2-dimethylcyclohexane and its effects on reproduction and development. The test
substance was administered to male rats (12 males/dose) for 14 days before mating and throughout
the mating period until the day before necropsy (42 days) and to female rats (12 females/dose) for
14 days before mating and throughout the mating and gestation periods until day 4 of lactation (41 to
45 days) at doses of 0 (vehicle: 0.5w/v% carboxymethylcellulose sodium solution containing 0.1v/v%
Tween 80), 100, 300, and 1,000 mg/kg bw/day. Furthermore, a 14-day recovery period was set for eachfive males from the control and the 1,000 mg/kg bw/day groups. Five additional females at 0 and 1,000
mg/kg bw/day were dosed with the test substance for 42 days without mating and were examined aftera 14-day recovery period.
No effects were observed from the administration of the test article in clinical signs, the functional
observation battery (FOB: detailed clinical signs, manipulative test, grip strength, motor activity), bodyweight, food consumption, urinalysis, blood chemistry, or organ weight. At the end of the administration
period, in hematological examination, low values of hemoglobin and hematocrit were found in males
in the 1,000 mg/kg bw/day group. Histopathological examination showed minimal hypertrophy ofcentrilobular hepatocytes in males in the 1,000 mg/kg bw/day group. At the end of the recovery period,
the changes observed were no longer observed. The changes in the hematological and histopathologicalexaminations at 1,000 mg/kg bw/day indicated that the NOAEL of repeated-dose toxicity is 300 mg/
kg bw/day in rats.

11

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

UUID: f51293f9-962e-4846-87f0-9e54ad3514ea
Dossier UUID:
Author:Date: 2022-12-15T11:20:22.622+09:00
Remarks:

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)

Data source
Reference
Reverse Mutation Test of perfluoro-1,2-dimethylcyclohexane / Ministry of Health, Labour and 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
GLP compliance
yesType of assay
bacterial reverse mutation assay
12

in vitro gene mutation study in bacteria

Test material
Specific details on test material used for the study- Name of test material (as cited in study report): Perfluoro-1, 2-dimethylcyclohexane- Lot No.: 11821MD (Sigma-Aldrich corporation)- Purity: >97.7%- Solubility: soluble in water, slightly soluble in 0.5% carboxymethyl cellulose, and insoluble in dimethyl
sulfoxide and acetone.
- Physical state: Colorless liquid- Storage condition of test material: in cool (1-8 degree C)

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, 250, 5000 μg /plate
Maximum concentration was established based on the result of the preliminary test at concentration
up to 5000 ug/plate. In this test, the growth inhibition was not observed at 5000 μg/plate (highest
concentration) for all strains with and without S9 mix.
Vehicle / solvent
water
Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes
True negative controlsnoPositive controls
yes
Positive control substanceother: without S9 mix: sodium azide (TA 1535) without S9 mix:2-(2-Furyl)-3-(5-nitro -2-furyl)acry
lamide (TA100, TA98, WP2uvrA), without S9 mix: 9-Aminoacridine (TA1537) with S9 mix: 2-(2Furyl)-3-(5-nitro-2-furyl)acrylamide (all strains)13

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
trueSpecies / strain
S. typhimurium 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
Positive controls validity
valid
Key result
trueSpecies / strain
S. typhimurium TA 1535
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
14

Vehicle controls validity
valid
Positive controls validity
valid
Key result
trueSpecies / strain
S. typhimurium TA 1537
bacteria
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Positive controls validity
valid
Key result
trueSpecies / strain
S. typhimurium TA 98
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
Positive controls validity
valid
Key result
trueSpecies / strain
E. coli WP2 uvr A pKM 101
bacteria
Metabolic activation
with and without
15

Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity
valid
Positive controls validityvalid

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/PDF306-98-9e.pdf

Applicant's summary and conclusion
Conclusions
Genotoxic effects:
With metabolic activation: Negative
Without metabolic activation: Negative
Executive summary
In a bacterial reverse mutation assay using S. typhimuriumTA100, TA1535, TA98, and TA1537,
and E. coli WP2uvrA/pKM101 (OECD TG 471), negative results were obtained for perfluoro-1,2dimethylcyclohexane with or without metabolic activation.

16

ENDPOINT_STUDY_RECORD: Genetic toxicity in vitro.002

UUID: 0f0f075e-9ba7-467c-8d1b-8639c730a2d3
Dossier UUID:
Author:Date: 2022-12-15T11:21:11.797+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 perfluoro-1,2-dimethylcyclohexane Cultured Chinese
Hamste / Ministry of Health, Labour and Welfare (MHLW), Japan / study report
Data accessdata published

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

Test material
17

Specific details on test material used for the study- Name of test material (as cited in study report): Perfluoro-1, 2-dimethylcyclohexane- Lot No.: 11821MD (Sigma-Aldrich corporation)- Purity: >97.7%- Solubility: soluble in water, slightly soluble in 0.5% carboxymethyl cellulose, and insoluble in dimethyl
sulfoxide and acetone.
- Physical state: Colorless liquid- Storage condition of test material: in cool (2-8 degree C)

MethodSpecies / strain
Species / strain / cell typeother:
Details on mammalian cell type (if applicable)
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
0, 1000, 2000, 4000 μg/mL
Cell-growth inhibition test was conducted up to the limited concentration of 4000 μg/mL (10 mM)
Cell growth inhibition was not observed for the short-term (+/-S9 mix) and continuous treatment
Vehicle / solvent
water
Controls
Untreated negative controlsnoNegative solvent / vehicle controls
yes
True negative controlsnoPositive controls
yes
Positive control substancebenzo(a)pyrene(with S9 mix)
mitomycin C(without S9 mix)
Details on test system and experimental conditions
METHOD OF APPLICATION: Exposure duration: [short-term treatment]:6 hrs + 18 hr, [continuous
treatment]: 24h
NUMBER OF CELLS EVALUATED: 200 cells /concentration (100 cells/plate x 2)
18

Plates/test: 4
Evaluation criteria
The followings were judged as positive: 1) total chromosomal aberrations increased >10% andconcentration dependent increase was observed, 2) total chromosomal aberrations increased >5%
and reproducible.
Statistics
not used

Results and discussion
Test results
Key result
trueSpecies / strainother: Chinese hamster lung(CHL/IU) cells
Metabolic activation
with and without
Genotoxicity
negative
Cytotoxicity / choice of top concentrations
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity
valid
Positive controls validityvalid

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/PDF306-98-9f.pdf

Applicant's summary and conclusion
Conclusions
without S9 mix: negativewith S9 mix: negative
Executive summary
In an in vitro chromosomal aberration test using CHL/IU cells (OECD TG 473), negative results wereobtained with or without metabolic activation.

19

Toxicity to reproduction
ENDPOINT_STUDY_RECORD: Toxicity to reproduction.001

UUID: 16b6ebf8-e3bd-4d69-9a8b-de5c461b349d
Dossier UUID:
Author:Date: 2022-12-16T16:30:34.204+09:00
Remarks:

Administrative data
Endpoint
reproductive toxicity, other A combined repeated dose/reproductive developmental toxicity study
Type of information
experimental study
Adequacy of study
key study
Robust study summary
false
Used for classification
false
Used for SDS
false
Reliability
1 (reliable without restriction)
Rationale for reliability incl. deficiencies
guideline study
Reliability 1
Cross-reference
Reason / purpose for cross-reference
reference to same study
Related information
OECD / Repeated dose toxicity: oral / Repeated dose toxicity: oral.001 / perfluoro-1,2dimethylcyclohexane / 1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexa

Data source
Reference
A combined repeated dose/reproductive developmental toxicity study
dimethylcyclohex / Ministry of Health, Labor and Welfare, Japan / study report
Data accessdata published

Materials and methods
20ofperfluoro-1,2-

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
Specific details on test material used for the study- Name of test material (as cited in study report): Perfluoro-1, 2-dimethylcyclohexane- Analytical purity: 97%- Storage condition of test material: at a room temperature (15-23°C) , protected from light- Stability under test conditions: The stability of test material was identified by analysis of the remainder.

Test animals
Speciesrat
Strainother: Crl:CD(SD)Sexmale/female
Details on test animals or test system and environmental conditions
TEST ANIMALS- Source: Charles River Japan, Inc., Atsugi Breeding Center.
- Age at study initiation: 11 weeks old- Weight at study initiation: Male: 397 g (364 -442 g), Female: 244 g (226-268 g)- Housing: Animals were individually housed in bracket-type metallic wire-mesh cages (254W × 350D
× 170H mm), from gestation day 17 to lactation day 4, Dams were bred individually or with individual
littermates in plastic cages (340W x 400D x 185H mm) and bedding.
- Diet: Solid feed (NMF: Oriental Yeast Co., ltd.) was given ad libitum.
- Water: Tap water was given ad libitum.
- Acclimation period: 20 days
ENVIRONMENTAL CONDITIONS- Temperature (°C): 23±3 (actual temperature: 21-28°C)- Humidity (%): 50±20% (actual humidity: 33-47%)- Air changes (per hr): 10-15- Photoperiod (hrs dark / hrs light): 12 hr dark/12 hr light (light: 7:00~19:00)

21

Administration / exposure
Route of administration
oral: gavage
Vehicleother: 0.5 w/v% CMC-Na solution mixed containing 0.1 v/v% Tween 80
Details on exposure- Amount of vehicle (if gavage): 5 mL/kg- Dosing volume: 5 mL/kg
Analytical verification of doses or concentrations
yes
Duration of treatment / exposure(P) Males: 42 days including 14 days pre-mating(P)Females: 41-45 days including 14 days pre-mating, mating and gestation periods and the days until
day 4 of lactation
Female (no mating, satellite group): 42 days

Frequency of treatment
Once/day, 7 days/week
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
Main group:12 animals/sex/dose
Satellite (Recovery) group: 5 males/dose and 5 females/dose (0 and 1000 mg/kg bw/day)Control animals
yes, concurrent vehicle
Details on study design- Dose selection rationale: Based on the results of a 14-day preliminary study, the highest dose of1000 mg/kg bw/day was selected as an expected obvious toxic dose, and the lowest dose of 100 mg/
kg bw/day was selected as an expected no toxic dose. The middle dose levels of 300 mg/kg bw/day
were selected.
- Rationale for animal assignment (if not random): Body weight-balanced randomization22[14-day preliminary study]
A 14-day repeated dose oral toxicity test (Crl:CD(SD) rats, doses: 0, 100, 300 or 1000 mg/kg bw/day). No death was observed even at 1000 mg/kg bw/day, and no effects were observed in any groups
except for a slightly decreased in MCH in males at 300 and 1000 mg/kg bw/day.

Examinations
Parental animals: Observations and examinations
CAGE SIDE OBSERVATIONS: Yes- Time schedule: 3 times/day (before administration, immediately after administration and 2 hours
after administration) during the administration period. Once a day during the recovery period.
DETAILED CLINICAL OBSERVATIONS: Yes- Time schedule:
Male main group: once before the start of administration, once every weekly during the administration
and recovery periods.
Female main group: once before the start of administration, days 1, 7, 14 and 20 of gestation, and day
4 of lactation.
Male and female recovery group: once before the start of administration, once every weekly during theadministration and recovery periods.
BODY WEIGHT: Yes- Time schedule for examinations:
Males in the main groups were weighed on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 ofadministration and on the day of necropsy, and 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 in addition to the meas
urement days for males in the main groups.
Females in the main groups were weighed on days 1, 4, 8, 11 and 15 of administration (uncopulated
animals were weighed on day 18 of administration as well), days 0, 4, 7, 11, 14, 17 and 20 of gestation, days 0 and 4 of lactation and the day of necropsy.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):- Food consumption: Yes
Measurement of food consumption was conducted on all animals at the following frequencies:
males in the main groups on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration; males and fem
ales in the recovery groups on days 1, 4, 8, 11 and 14 of recovery in addition to the measurement da
ys for males in the main groups; and females in the main groups on days 1, 4, 8, 11 and 15 of admini
stration, days 1, 4, 7, 11, 14, 17 and 20 of gestation and days 2 and 4 of lactation.
OPHTHALMOSCOPIC EXAMINATION: No
HAEMATOLOGY: Yes- Time schedule for collection of blood: At the end of administration period, or at the end of recovery
period in both sexes- Anaesthetic used for blood collection: ether- Animals fasted: Yes- How many animals: 5 animals/sex/group- Parameters examined: red blood cell count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, reticulocyte p
ercentage, platelet count, white blood cell count, differential white blood cell count, absolute numberof each white blood cell, prothrombin time, activated partial thromboplastin time, fibrinogen.
CLINICAL CHEMISTRY: Yes- Time schedule for collection of blood: At the end of administration period, or at the end of recovery
period in both sexes- Animals fasted: Yes- How many animals: 5 animals/sex/group23

- Parameters checked: ALP, total cholesterol, triglyceride, phospholipids, total bilirubin, glucose, bloodurea nitrogen, creatinine, sodium, potassium, chloride, calcium, inorganic phosphorus, total protein,
albumin, A/G ratio, AST (GOT), ALT (GPT), LDH, γ-GTP
URINALYSIS OF MALES: Yes- Time schedule for collection of urine: final week of administration (days 39 to 40 of administration)and in the final week of recovery (days 11 to 12 of recovery)- Metabolism cages used for collection of urine: Yes
A urine collector to collect four-hour urine samples under fasting but ad libitum drinking conditions,followed by collection of 20-hour urine samples under ad libitum feeding and drinking conditions.
- How many animals: 5 animals/group- Parameters checked: pH, protein, ketones, glucose, occult blood, bilirubin, urobilinogen, color, sedime
nt, urine volume (4-hour volume), osmotic pressure, urine volume (20-hour volume), water intake (24hour volume)- Time schedule for examinations: Manipulative test and measurements of grip strength and motor activity were conducted on 5 animals per group with the following frequencies: males in the main grou
ps were examined in the final week of administration (day 37 of administration), females in the maingroups on day 4 of lactation (day 41 to day 44 of administration) after necropsy of F1 pups, and ma
les and females in the recovery groups in the final week of administration (day 37 of administration)and in the final week of recovery (day 9 of recovery).
- Battery of functions tested:
1) Open field observation. Arousal, gait, posture, tremor, convulsion, rearing count, defecation (defecation count, urination), stereotypy (grooming, circling, etc.), abnormal behavior (self-biting, ba
ckward walking, etc.)
2) Manipulative Test. Auditory response, approach response, touch response, tail pinch response, p
upillary reflex, aerial righting reflex, landing foot splay
3) Measurement of Grip Strength. Following manipulative test, grip strength of forelimb and hind limb
was measured by CPU gauge MODEL-9502A (AIKOH Engineering Co., Ltd.).
4) Measurement of Motor Activity. Following measurement of grip strength, motor activity wasmeasured by a motor activity sensor for experimental animals NS-AS01 (Neuro Science, Inc). The
measurement was conducted for 1 hour, and measured values at 10-minute intervals and from 0 to 60
minutes were collected.
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 administration until the day copulation was confirmed.
During the pre-mating administration period, vaginal smear pictures were classified as proestrus,
estrus, metestrus or diestrus and examined for the frequency of estrus and interval between estruses(estrous cycle). During the mating period, vaginal smears were examined for the presence of sperm.
Sperm parameters (parental animals)
Parameters examined in all P male parental generations: testis weight, epididymis weight, histopatho
logical examinations for testes, epididymides, seminal vesicle and ventral prostate.
Litter observations
PARAMETERS EXAMINED: The following parameters were examined in F1 offspring: Number and sexof pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, and weight gain.
GROSS EXAMINATION OF DEAD PUPS: Yes, for external and internal abnormalities.
Postmortem examinations (parental animals)
METHOD OF SACRIFICED: All animals were sacrificed by exsanguination under ether anesthesia.
SACRIFICE: Male animals: On next day after the last administration (Day 43), Maternal animals: on
Day 4 of lactation, and Male recovery and female satellite animals: on Day 14 of recovery.

24

GROSS PATHOLOGY: Yes
ORGAN WEIBHT: Yes [brain, thyroids including parathyroids), adrenal gland, thymus, spleen, heart,
liver, kidney, testis, epididymis]
HISTOPATHOLOGY: Yes, [cerebrum, cerebellum, pituitary, spinal cord (thoracic), sciatic nerve, thyroid, parathyroid, adrenal glands, thymus, spleen, submandibular lymph nodes, mesenteric lymph nodes,
heart, lung (including bronchial), stomach, duodenum, jejunum, ileum, cecum, colon, rectum, liver, kidney, bladder, testis, epididymis, ovary, uterus, prostate, seminal vesicles, sternum and femur (includingbone marrows), macroscopic lesions, and parts for identification (auricles)]Postmortem examinations (offspring)
SACRIFICE- The F1 offspring were sacrificed at 4 days of age.
GROSS NECROPSY- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, andabdominal viscera.
HISTOPATHOLOGY / ORGAN WEIGTHS- Not examined.
Statistics
The data were analyzed for homogeneity of variance by the Bartlett test. If variances were
homogeneous, data was analyzed by the Dunnett test, whereas heterogeneous data was analyzed by
the Dunnett type mean rank test (p<0.05, two-sided).
In the recovery test, these values of two groups were analyzed by F test. If variances were homogeneo
us, data was analyzed by the Student t-test, whereas heterogeneous data was analyzed by the AspinWelch t-test (p<0.05, two-sided).
Reproductive indices
Each parameter was determined by the following equations:
Copulation index (%) = (No. of copulated animals/No. of co-housed animals) × 100
Fertility index (%) = (No. of pregnant females/No. of copulated females) × 100
Insemination index (%) = (No. of pregnant females/No. of copulated males) × 100
Duration of gestation (days) = day 0 of lactation – day 0 of gestation
Delivery index (%) = (No. of females delivered liveborn pups/No. of pregnant females) × 100
Implantation index (%) = (No. of implantation sites/No. of corpora lutea) × 100
Stillborn index (%) = (No. of stillborn pups/Total No. of pups born) × 100
Liveborn index (%) = (No. of liveborn pups/Total No. of pups born) × 100
External abnormalities (%) = (No. of pups with external abnormalities/No. of liveborn pups) × 100
Sex ratio = No. of liveborn male pups/(No. of liveborn male pups + No. of liveborn female pups)
Offspring viability indices
Viability index (%) = (No. of live pups on day 4/No. of live pups on day 0) × 100

Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
Clinical signs
no effects observed25Mortality
no mortality observed
Body weight and weight changes
no effects observed
Food consumption and compound intake (if feeding study)no effects observed
Food efficiency
not examined
Water consumption and compound intake (if drinking water study)
not examined
Ophthalmological findings
not examined
Haematological findings
effects observed, treatment-related
Description (incidence and severity)
See 7.5.1
Clinical biochemistry findings
no effects observed
Urinalysis findings
no effects observed
Behaviour (functional findings)
no effects observed
Immunological findings
not examined
Organ weight findings including organ / body weight ratios
no effects observed
Gross pathological findings
no effects observed
Neuropathological findings
not examined
Histopathological findings: non-neoplastic
effects observed, treatment-related
Description (incidence and severity)
See 7.5.1
Histopathological findings: neoplastic
not examined

Reproductive function / performance (P0)
Reproductive function: oestrous cycle
no effects observed
Reproductive function: sperm measures
no effects observed
26

Reproductive performanceno effects observed

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

mg/kg bw/day (actual dose received)

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

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

Effect levels (F1)
Key result
true
Dose descriptor
NOAEL
Generation
F1
Effect level1000

mg/kg bw/day (actual dose received)27Based on
test mat.
Sexmale/female
Basis for effect level
other: No effects on development

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/PDF306-98-9d.pdf

Applicant's summary and conclusion
Executive summary
In the above-described OECD TG 422 study, no effects were observed for reproduction or development.
The NOAEL for rat reproduction/developmental toxicity of perfluoro-1, 2-dimethylcyclohexane wasestimated to be 1, 000 mg/kg bw/day (the highest dose tested).

28

DOMAIN
Substance
SUBSTANCE: perfluoro-1,2-dimethylcyclohexane
UUID: ae31cb58-6a0d-42e4-b092-1d1cb9ec9989
Dossier UUID:
Author:Date: 2022-12-16T16:30:50.963+09:00
Remarks:
Substance nameperfluoro-1,2-dimethylcyclohexane
Legal entity
National Institute of Health Sciences / Kawasaki / Japan

Identification of substance
Reference substance
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexanebis(trifluoromethyl)cyclohexane / 306-98-9 / 206-195-0
EC number

EC name206-195-0

EC Inventory

CAS number

CAS name

306-98-9
IUPAC name
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexane

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

29

/

1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-

References
Reference Substances
REFERENCE_SUBSTANCE: 1,1,2,2,3,3,4,4,5,6decafluoro-5,6-bis(trifluoromethyl)cyclohexane
UUID: ECB5-21f79ee5-31d6-493a-bdef-04069e6c5f5b
Dossier UUID:
Author:Date: 2019-02-15T14:29:53.000+09:00
Remarks:
Reference substance name
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexane
IUPAC name
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexane

Inventory
Inventory number
Inventory name
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexane
Inventory
EC Inventory
Inventory number
206-195-0
CAS number
306-98-9
Molecular formula
C8F16
Description
CAS number
306-98-9

Synonyms
Synonyms
Identity
1,1,2,2,3,3,4,4,5,6-decafluoro-5,6-bis(trifluoromethyl)cyclohexane

Molecular and structural information
30

Molecular formula
C8F16
Molecular weight400.0601
SMILES notation
FC(F)(F)C1(F)C(F)(F)C(F)(F)C(F)(F)C(F)(F)C1(F)C(F)(F)F
InChl
InChI=1/C8F16/c9-1(7(19,20)21)2(10,8(22,23)24)4(13,14)6(17,18)5(15,16)3(1,11)12
Structural formula31Literatures
LITERATURE: A combined repeated dose/reproductive
developmental toxicity study of perfluoro-1,2dimethylcyclohexane cyclohexane by oral administrationin rats.
UUID: b50cc229-f562-4295-b872-8794524835ce
Dossier UUID:
Author:Date: 2022-12-15T11:14:48.001+09:00
Remarks:

General information
Reference Typestudy report
Title
A combined repeated dose/reproductive developmental toxicity study of perfluoro-1,2-dimeth
ylcyclohexane cyclohexane by oral administration in rats.
Author
Ministry of Health, Labor and Welfare, Japan
Bibliographic source
Japan Existing Chemical Data Base (JCDB)
Testing facility
Bozo Research Center Inc.
Report number
R-967

32

LITERATURE: In Vitro Chromosomal Aberration Test ofon perfluoro-1,2-dimethylcyclohexane Cultured Chinese
Hamster Cells
UUID: 8ae74eb1-b9e0-40a3-9deb-604b590b5be4
Dossier UUID:
Author:Date: 2019-02-15T14:16:35.000+09:00
Remarks:

General information
Reference Typestudy report
Title
In Vitro Chromosomal Aberration Test of on perfluoro-1,2-dimethylcyclohexane Cultured Chinese
Hamster Cells
Author
Ministry of Health, Labour and Welfare (MHLW), Japan
Bibliographic source
http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp

33

LITERATURE: Reverse Mutation Test of perfluoro-1,2dimethylcyclohexane
UUID: 5ae4c893-1e85-46a7-ab87-7a22f378cfe1
Dossier UUID:
Author:Date: 2019-02-15T10:58:20.000+09:00
Remarks:

General information
Reference Typestudy report
Title
Reverse Mutation Test of perfluoro-1,2-dimethylcyclohexane
Author
Ministry of Health, Labour and Welfare (MHLW), Japan
Bibliographic source
http://dra4.nihs.go.jp/mhlw_data/jsp/SearchPageENG.jsp
Testing facility
Research Institute for Animal Science in Biochemistry and Toxicology (RIAS)

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