Perfluamine

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 421, Reproduction/Developmental Toxicity Screening Test (2016)

Version / remarks:

2016

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

PTPA

IUPAC Name:

PTPA

Details on test material:

- Name of test material (as cited in study report):TCR11-0027-1/1 (FC-3283)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3M Company, Batch 3435912
- Expiration date of the lot/batch: 07 July, 2021
- Purity test date: 12 December, 2017

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: NA, dosed neat

TREATMENT OF TEST MATERIAL PRIOR TO TESTING : None, dosed neat.

Test animals

Species:

rat

Strain:

Wistar

Details on species / strain selection:

The strain is recommended per OECD 421.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany or Charles River Laboratories France, L'Arbresle Cedex, France
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 10-12 weeks
- Weight at study initiation: Males: 250-350 g, Females: 200-250 g
- Fasting period before study: None
- Housing: On arrival and following the pretest (females only) and pre-mating period, animals will be group housed (up to 5 animals of the same sex and same dosing group together) in polycarbonate cages (Macrolon, MIV type, height 18 cm). During the mating phase, males and females will be cohabitated on a 1:1 basis in Macrolon plastic cages (MIII type, height 18 cm). During the post-mating phase, males will be housed in their home cage (Macrolon plastic cages, MIV type, height 18 cm) with a maximum of 5 males/cage. Females will be individually housed in Macrolon plastic cages (MIII type, height 18 cm). During the lactation phase, females will be housed in Macrolon plastic cages (MIII type, height 18 cm). Pups will be housed with the dam. The cages will contain appropriate bedding (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and will be equipped with water bottles. The housing conditions will be maintained unless deemed inappropriate by the Study Director and/or Clinical Veterinarian. The room(s) in which the animals will be kept will be documented in the study records. Animals will be separated during designated procedures/activities. Each cage will be clearly labeled with a color-coded cage card indicating Test Facility Study No., group, animal number(s), and sex.
- Diet (e.g. ad libitum): Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) will be provided ad libitum throughout the study, except during designated procedures. The feed is analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis are provided by the supplier and are on file at the Test Facility. It is considered that there are no known contaminants in the feed that would interfere with the objectives of the study.
- Water (e.g. ad libitum): Municipal tap water will be freely available to each animal via water bottles. Periodic analysis of the water is performed, and results of these analyses are on file at the Test Facility. It is considered that there are no known contaminants in the water that would interfere with
the objectives of the study.
- Acclimation period: At least 5 days.

DETAILS OF FOOD AND WATER QUALITY: See diet and water sections above.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24
- Humidity (%): 40-70
- Air changes (per hr): At least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 02 January,m 2019 To: 21 March, 2019

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

The test article was dosed neat via oral gavage.

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

- PREPARATION OF DOSING SOLUTIONS: The test item, MTDID 42916 was administered as received. An adequate amount of the test item was dispensed into daily aliquots, which were stored at room temperature.

- VEHICLE : None

Analytical verification of doses or concentrations:

no

Remarks:

The test item was used as received from the Sponsor; therefore, samples for dose formulation analysis were not collected by the Test Facility.

Details on analytical verification of doses or concentrations:

The test item was used as received from the Sponsor; therefore, samples for dose formulation analysis were not collected by the Test Facility.

Duration of treatment / exposure:

The test item (and Elix water for the control group) was administered to the appropriate animals by once daily oral gavage 7 days a week for a minimum of 28 days. Males were treated for 29 days, up to and including the day before scheduled necropsy. This included a minimum of 14 days prior to mating and during the mating period. Females that delivered were treated for 51-64 days, i.e. 14 days prior to mating (with the objective to cover at least two complete estrous cycles), the variable time to conception, the duration of pregnancy and at least 13 days after delivery, up to and including the day before scheduled necropsy. Females which failed to deliver were treated for 40-54 days.

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Elix water control (Group 1)

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Group 2

Doses / concentrations 3

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Group 3

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Group 4

No. of animals per sex per dose:

10

Control animals:

yes, sham-exposed

Details on study design:

- Dose selection rationale: Other repeated dose studies.
- Rationale for animal assignment (if not random): Random
- Rationale for selecting satellite groups: NA
- Post-exposure recovery period in satellite groups: NA

Positive control:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Throughout the study, animals were observed for general health/mortality and moribundity twice daily, in the morning and at the end of the working day. Animals were not removed from the cage during observation, unless necessary for identification or confirmation of
possible findings.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Clinical observations were performed once daily, beginning during the first administration of the test item and lasting throughout the dosing periods up to the day prior to necropsy. During the dosing period, these observations were performed directly after dosing. The time of onset, grade and duration of any observed sign was recorded. Signs were graded for severity and the maximum grade was predefined at 3 or 4. Grades were coded as slight (grade 1), moderate (grade 2), severe (grade 3) and very severe (grade 4). For certain signs, only its presence (grade 1) or absence (grade 0) was scored. In the data tables, the scored grades were reported, as well as the percentage of animals affected in summary tables.

BODY WEIGHT: Yes
- Time schedule for examinations: Animals were weighed individually on the first day of treatment (prior to dosing), and weekly thereafter. Mated females were weighed on Days 0, 4, 7, 11, 14, 17, and 20 post-coitum and during lactation on PND 1, 4, 7, and 13. Thirteen days after start of mating additional body weights were determined from females Nos. 59, 64 and 74 to check their pregnancy status because of the absence of evidence of mating. A terminal body weight was recorded on the day of scheduled necropsy.

FOOD CONSUMPTION: Food consumption was quantitatively measured weekly, except for males and females which were housed together for mating and for females without evidence of mating. Food consumption of mated females was measured on Days 0, 4, 7, 11, 14, 17, and 20 post-coitum
and during lactation on PND 1, 4, 7, and 13.

WATER CONSUMPTION: Subjective appraisal was maintained during the study, but no quantitative investigation was introduced as no effect was suspected.

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood:
Blood of F0-animals was collected on the day of scheduled necropsy. F0-males were fasted overnight with a maximum of 24 hours before blood sampling, but water was available. F0-females were not fasted overnight. Blood of F1-animals was collected on PND 4 and PND 14-16, if possible. This was performed in the necropsy room.
- Anaesthetic used for blood collection: Yes (Isofluorane)
- Animals fasted: Males were fasted, females were not
- How many animals:
All F0 animals and F1 animals at culling. On PND 14-16 separate blood samples were collected from two pups per litter (from one male and one female).
- Parameters checked: Serum T4 levels

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

OTHER: Estrous cycles were evaluated by examining the vaginal cytology of samples obtained by vaginal lavage. Daily vaginal lavage was performed for all females beginning 14 days prior to treatment (pretest period), the first 14 days of treatment and during mating until evidence of copulation
was observed. Vaginal lavage was continued for those females with no evidence of copulation until termination of the mating period. On the day of necropsy, a vaginal lavage was also taken to determine the stage of estrous.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes, All animals were subjected to a full post mortem examination, with special attention being paid to the reproductive organs. Necropsy procedures were performed by qualified personnel with appropriate training and experience in animal anatomy and gross pathology. A veterinary pathologist, or other suitably qualified person, was available. The numbers of former implantation sites were recorded for all paired females. In case no macroscopically visible implantation sites were present, non-gravid uteri were stained using the Salewski technique in order to detect any former implantation sites and the number of corpora lutea was recorded in addition.

HISTOPATHOLOGY: Yes

The following tissues were embedded in paraffin, sectioned, mounted on glass slides, and stained with hematoxylin and eosin:
All animals: Gross lesions/masses
All animals of Groups 1-4: Epididymis, coagulation gland, prostate gland, seminal vesicle gland, thyroid gland, ovaries, testes

Results and discussion

Results of examinations

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

One male (no.24, Group 3) showed abnormal gait during the last week of mating (fourth week of treatment). In the study files it was recorded that it did not properly used its hind legs. A few days before planned necropsy, veterinary inspection was conducted on this male. It was noted that it had its ears moved backwards and a rough fur, possible indications of pain. Moreover, a slight body weight loss was observed during the two weeks mating period. Furthermore, the male didn’t succeed in successful mating with its female partner. The cause of the injury of the hind legs was unknown, but might be related to the other findings in this male. It was considered in incidental finding that was not related to treatment. Alopecia noted in a single female (no. 56, Group 2) during the lactation phase occurred within the range of background findings to be expected for rats of this age and strain which are housed and treated under the conditions in this study.
In the absence of a dose-related trend and at the incidence observed, these findings in a single male and female were considered to be incidental findings and of no toxicological significance.

Mortality:

no mortality observed

Description (incidence):

No mortality occurred during the study period.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Body weights and body weight gain were considered to have been unaffected by treatment.
In high dose males, a slightly lower body weight gain was observed after treatment at 1000 mg/kg/day, achieving levels of statistical significance in several occasions when compared to controls. However, the average body weight gain in high dose animals was normal in comparison with the historical control data for rats of this strain and age (see Appendix 5), whereas that in control males was slightly higher than expected. Since the difference in mean body weights between control and high dose males did not exceed 3% during the study, no toxicological significance was attached to this finding. In high dose females at 1000 mg/kg/day, the mean body weights were slightly lower when compared to controls throughout the study, achieving a level of statistical significance on some occasions, i.e. Day 4 and 7 post coitum and Day 1 of lactation. However, during the study the relative difference in mean body weights varied from approximately 2% to 5% between controls and high dose females, indicating that the body weight gain was comparable between the two groups. Since the relative differences in mean body weights between controls and high dose females were similar at start and at the end of the study (i.e. approximately 3% lower), no toxicological significance was attached to this finding.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Food consumption before or after correction for body weight in the high dose group at 1000 mg/kg/day (Group 4) was similar to the control group (Group 1) over the treatment period.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

Serum levels of T4 in F0 males were considered not to be affected by treatment.

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Organ weights and organ to body weight ratios of treated animals were considered to be similar to those of control animals.

Gross pathological findings:

no effects observed

Description (incidence and severity):

Macroscopic observations at necropsy did not reveal any alterations that were considered to have arisen as a result of treatment.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

There were no test item-related microscopic observations.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

There were no test item-related microscopic observations.

Other effects:

no effects observed

Description (incidence and severity):

Length and regularity of the estrous cycle were considered not to have been affected by treatment. Mating index was considered not to be affected by treatment. All Group 1 and 4 females showed evidence of mating. Precoital time was considered not to be affected by treatment. All Group 1 and 4 females showed evidence of mating within 4 days after start of mating. Number of implantation sites was considered not to be affected by treatment. Fertility index was considered not to be affected by treatment.

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food consumption and compound intake

gross pathology

histopathology: neoplastic

histopathology: non-neoplastic

mortality

organ weights and organ / body weight ratios

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Based on the results of the study, the No Observed Adverse Effect Level (NOAEL) for MTDID 42916 is 1000 mg/kg/day.

Executive summary:

The reproductive/developmental and repeated dose toxicity of the test article was evaluated in male and female Wistar Han rats.  The study was conducted according to OECD 421 in compliance with OECD GLP regulations.  Rats (10/sex/dose) were dosed with 0 (control), 100, 300, or 1000 mg/kg/day via oral gavage for a minimum of 28 days.  The following parameters and end points were evaluated in this study: mortality/moribundity, clinical signs, body weight and food consumption, estrous cycle determination, measurement of thyroid hormone T4 (F0-males), gross necropsy findings, organ weights and histopathologic examinations.  In addition, the following reproduction/developmental parameters were determined: mating and fertility indices, precoital time, number of implantation sites, gestation index and duration, parturition, maternal care, sex ratio and early postnatal pup development, mortality, clinical signs, body weights, sex, anogenital distance, areola/nipple retention and macroscopy, measurement of thyroid hormone T4 (PND 14-16 pups)).  Due to a dosing error occurring during the pre-mating period of the study, the animals of low and mid dose groups had received at least five 5 times less than intended. The results of these two groups were, therefore, excluded from interpretation. No parental toxicity was observed at the highest dose tested (1000 mg/kg/day).  No adverse treatment-related changes were noted in any of the parameters investigated in this study (clinical appearance, body weight, food consumption, T4 thyroid hormone levels (in males only), macroscopic examination, organ weights, and microscopic examination.  No treatment-related changes were noted in any of the reproductive parameters investigated (mating and fertility indices, precoital time, number of implantations, estrous cycle, spermatogenic profiling, and histopathological examination of reproductive organs).  No treatment-related changes were noted in any of the developmental parameters investigated in this study (gestation, viability and lactation indices, duration of gestation, parturition, sex ratio, maternal care and early postnatal pup development consisting of mortality, clinical signs, body weight, anogenital distance, areola/nipple retention, T4 thyroid hormone levels and macroscopic examination).  Based on the results of the study, the parental and reproductive/developmental No Observed Adverse Effect Level (NOAEL) is 1000 mg/kg/day.