Potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulphonate

Toxicity to reproduction

Administrative data

Endpoint:

two-generation reproductive toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.3800 (Reproduction and Fertility Effects)

Version / remarks:

1998

Deviations:

no

Remarks:

No deviations occurred that adversely affected the outcome or interpretation of the study.

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulphonate

EC Number:

249-616-3

EC Name:

Potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulphonate

Cas Number:

29420-49-3

Molecular formula:

C4HF9O3S.K

IUPAC Name:

potassium 1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonate

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Lots: 1, 2, and 3
- Expiration date of the lot/batch: 31 August, 2003
- Purity test date: 31 August, 2001

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: The test article was stored at room temperature
- Stability under test conditions: Samples of each prepared bulk test substance was retained on the day after the last day of treatment and analyzed to ensure proper test article dosing.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test article was suspended in 0.1% carboxymethyl cellulose (CMC) in deionized water.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on species / strain selection:

The Crl:CD(SD)IGS BR VAF/Plus rat was selected as the Test System because of known response to toxic effects on reproductive capacity and history of use as a rodent species in these evaluation.

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Kingston, New York
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: (P) 6 wks; (F1) 3 wks
- Weight at study initiation: (P) Males: 128-157 g; Females: 118-144 g
- Fasting period before study: None
- Housing: P generation rats were individually housed in stainless steel, wire-bottomed cages, except during the cohabitation and postpartum periods. During cohabitation, each pair of rats was housed in the male rat's cage. Beginning no later than DG 20, female rats were individually housed in nesting boxes. Each dam and delivered litter was housed in a common nesting box during the postpartum period.

After weaning, the Fl generation rats were individually housed before cohabitation, housed in pairs (one male rat per female rat) during cohabitation, and individually housed after cohabitation. The same type of caging was used as described for the P generation rats. Beginning no later than DG 20, female rats were individually housed in nesting boxes. Each dam and delivered litter was housed in a common nesting box during the postpartum period.

- Diet (e.g. ad libitum): Rats were given Certified Rodent Diet #5002 (PMI Nutrition International, St. Louis, Missouri) available ad libitum from individual feeders throughout the study.
- Water (e.g. ad libitum): Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad Zib.iturn from individual water bottles attached to the cages and/or from an automatic watering access system. Chlorine was added to the processed water as a bacteriostat.
- Acclimation period: 7 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26
- Humidity (%): 30-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 19 December, 2000 To: 07 September, 2001

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

0.1% CMC in deionized water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test article was suspended in 0.1% CMC in deionized water

VEHICLE
- Justification for use and choice of vehicle (if other than water): 0.1% carboxymethyl cellulose (CMC) in deionized water
- Concentration in vehicle: The test article was prepared in vehicle so that it could be dosed at 0 (control), 30, 100, 300, and 1000 mg/kg/day at a volume of 10 mL/kg.
- Amount of vehicle (if gavage): 10 mL/kg.
- Lot/batch no. (if required): 49H1332

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: vaginal plug / sperm in vaginal smear referred to as Day 0 of pregnancy
- After successful mating each pregnant female was caged: Individually in nesting boxes.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The 3, 10, 30 and 66.7 mg/mL samples were all found to be within 2 10% of the target concentrations; the 100 mg/mL samples were 76.6% (first preparation) and 99.1% (last preparation) of target.

Duration of treatment / exposure:

P generation rats (30/sex/dose) received 0 (vehicle), 30, 100, 300, or 1000 mg/kg/day of T-7485 via oral gavage at a dose volume of 10 mL/kg beginning at approximately 6 weeks of age (at least 70 days before cohabitation) and continuing until the day before sacrifice (males: after completion of cohabitation period; females: DL 22). All F1 pups were weaned on DL 22 and were randomly selected for continued evaluation. F1 pups (30/sex/dose) received 0 (vehicle), 30, 100, 300, or 1000 mg/kg/day of T-7485 via oral gavage beginning at weaning (approximately 70 days before cohabitation) and continuing until the day before sacrifice (males: after completion of cohabitation period; females: DL 22).

Frequency of treatment:

Daily during treatment-appropriate times described in the duration section.

Details on study schedule:

- F1 parental animals not mated until approximately 10 weeks after from the F1 litters.
- Selection of parents from F1 generation when pups were approximately 22 days of age.
- Age at mating of the mated animals in the study: Approximately 13 weeks

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

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

Remarks:

Vehicle control (0.1% CMC in deionized water)

Doses / concentrations 2

Dose / conc.:

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

Doses / concentrations 3

Dose / conc.:

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

Doses / concentrations 4

Dose / conc.:

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

Doses / concentrations 5

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

30

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: Doses were selected based on a 28-day repeat dose oral toxicity study. The highest dosage was expected to produce some toxicity. The intermediate dosage levels were expected to produce minimal observable toxic effects, while the lowest dosage level was expected to produce no evidence of either systemic or reproductive toxicity [no-observable-adverse-effect level (NOAEL)] .
- Rationale for animal assignment: Random
- Other:

Positive control:

None

Examinations

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily
- Cage side observations checked: Viability

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: 60 minutes after dosage administration.

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights of the male rats were recorded weekly during the acclimation and dosage periods and on the day of sacrifice. Body weights of the female rats were recorded weekly during the acclimation, precohabitation and cohabitation periods, on DGs 0,7, 10, 14, 18, 21 and 25 (if necessary) and on DLs 1,5,8, 11, 15 and 22 (terminal body weight).

FOOD CONSUMPTION: Feed consumption values for the male rats were recorded weekly during the acclimation and dosage periods. Feed consumption values for the female rats were recorded weekly during the acclimation and precohabitation periods, on DGs 0,7, 10, 14, 18,21 and 25 (if necessary) and on DLs 1,5,8, 11, and 15. During cohabitation, when two rats occupied the same cage with one feed jar, replenishment of feed jars was documented but individual values were not recorded or tabulated. Because pups begin to consume maternal feed on or about DL 15, feed consumption values were not tabulated after DL 15.

WATER CONSUMPTION: No data

Oestrous cyclicity (parental animals):

P: Estrous cycling was evaluated daily by examination of vaginal cytology beginning 21 days before the scheduled cohabitation period and continuing until spermatozoa were observed in a smear of the vaginal contents andor a copulatory plug was observed in situ during the cohabitation period. On the day of scheduled sacrifice, an examination of vaginal cytology was performed to determine the stage of estrous cycle.

F1: Estrous cycling was evaluated daily by examination of vaginal cytology beginning 21 days before the scheduled cohabitation period and continuing until spermatozoa were observed in a smear of the vaginal contents and/or a copulatory plug was observed in situ during the cohabitation period. On the day of scheduled sacrifice, an examination of vaginal cytology was performed to determine the stage of estrous cycle.

Sperm parameters (parental animals):

Parameters examined in P/F1 male parental generations: testis weight, epididymis weight, sperm count in testes, sperm count in epididymides, sperm motility, sperm morphology.

Litter observations:

STANDARDISATION OF LITTERS
Litters were not culled during the lactation period because random selection of pups for culling could have resulted in potential biases in pup viabilities and body weight gains during this period. All F1 generation pups were weaned on DL, 22 based on observed growth and viability of these pups. At weaning, a table of random units was used to select 30 male and 30 female F1 generation pups per group, resulting in a total of 300 rats (150 rats per sex) chosen for continued evaluation. At least one male pup and one female pup per litter, when possible, were selected.

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring: F: Litters were examined after delivery to identify the number and sex of pups, stillbirths, live births and gross external alterations. DL 1 was defined as the day of birth and was also the first day on which all pups in a litter were individually weighed (pup body weights were recorded after all pups in a litter were delivered and groomed by the dam). Each litter was evaluated for viability at least twice each day of the 22-day postpartum period. Dead pups observed at these times 'were removed from the nesting box. F1 generation pups in each litter were counted once daily. Physical signs (including variations from expected lactation behavior and gross external alterations) were recorded for the pups each day. Pup body weights were recorded on DLs 1 (liveborn pups), 5 , 8 , 15 and 22.

F2: Litters were examined after delivery to identify the number and sex of pups, stillbirths, live births and gross external alterations. Each litter was evaluated for viability at least twice each day of the 22-day postpartum period. Dead pups observed at these times were removed from the nesting box. F1 generation pups in each litter were counted once daily. Physical signs (including variations from expected lactation behavior and gross external alterations) were recorded each day. Pup body weights were recorded on DLs 1 (liveborn pups), 5,8, 15 and 22.

GROSS EXAMINATION OF DEAD PUPS:
Yes, for external and internal abnormalities; possible cause of death was attempted to be determined for pups born or found dead.

Postmortem examinations (parental animals):

SACRIFICE
- Male animals: All surviving male rats were sacrificed after completion of the cohabitation period days of study (DSs) 122 to 123 for the P generation rats and DPs 129 to 141 for the Fl generation rats.

- Maternal animals: All surviving female parental animals were sacrificed on DL22.

GROSS NECROPSY
- Gross necropsy included an initial physical examination of external surfaces and all orifices, as well as an internal examination of tissues and organs in situ. The following were examined: external and internal portions of all hollow organs; the external surfaces of the brain and spinal column; the nasal cavity and neck with associated organs and tissues; the thoracic, abdominal and pelvic cavities with associated organs and tissues; and the musculo/skeletal carcass. The lungs of the Fl generation male rats and P and F1 generation female rats were perfused with neutral buffered 10% formalin and the urinary bladder of postweaning rats was inflated with neutral buffered 10% formalin.

HISTOPATHOLOGY / ORGAN WEIGHTS
The following organs were individually weighed and organ-to-body weight and organ-to brain weight ratios were calculated: brain, kidneys, spleen, ovaries, testes, thymus, liver, adrenal glands, pituitary, uterus with oviduc:ts and cervix, left epididymis (whole and cauda), right epididymis, prostate and semirial vesicles (with coagulating glands and with and without fluid)'. The following tissues were retained in neutral buffered 10% formalin for possible histological evaluation: pituitary, liver, adrenal glands, kidneys, vagina, uterus with oviducts, cervix and ovaries, left testis (initially fixed in Bouin's solution for 48 to 96 hours), seminal vesicles (with coagulating gland), right epididymis and prostated. Histological examination was performed on tissues from ten randomly selected rats per sex from the control and high dosage groups. All gross lesions were examined histologically. In addition to gross lesions such as atrophy or tumors, testicular histopathological examination was conducted to identify treatment-related effects such as retained spermatids, missing germ cell layers or types, multinucleated giant cells or sloughing of spermatogenic cells into the lumen. Examination of the intact epididymis included the caput, corpus and cauda (accomplished by evaluation of a longitudinal section) and conducted in order to identify such lesions as sperm granulomas, leukocytic infiltration (inflammation), aberrant cell types within the lumen, or the absence of clear cells in the cauda epididymal epithelium. Reproductive organs of the rats suspected of reduced fertility (e.g., those rats that failed to sire healthy offspring, or for which estrous cyclicity or sperm number, motility or morphology were affected) were subjected to histological evaluationa.

Postmortem examinations (offspring):

SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

Reproductive indices:

Estrous cycling, duration of gestation, fertility index, gestation index, number and sex of offspring per litter, number of implantation sites, litter size, sex ratio, sperm count and motility.

Offspring viability indices:

Fertility index, lactation index, percent survivval.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Males treated at 1000 mg/kg/day exhibited significant increases in the incidences of excess salivation, perioral substance, chromorhinorrea, and urine-stained abdominal fur. One or two males treated at 300 mg/kg/day exhibited excess salivation and perioral substance that were considered treatment-related. Females treated at 1000 mg/kg/day exhibited a significant increase in the incidence of dried or wet red perioral substance and excess salivation during precohabitation and gestation.

Dermal irritation (if dermal study):

not examined

Mortality:

mortality observed, non-treatment-related

Description (incidence):

One male treated at 1000 mg/kg/day was found dead on Day 55 as a result of an intubation accident. All other P generation animals survived until scheduled termination.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

At 1000 mg/kg/day, male body weight gains were significantly reduced on Days 43 to 50 and 50 to 57, and significant body weight loss occurred on Days 105 to 112. Body weights and body weight gains of females during precohabitation, gestation, and lactation were unaffected by T-7485 doses as high as 300 mg/kg/day. Terminal body weights of males and females were unaffected by test article administration.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no abnormal changes in absolute or relative food consumption values.

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:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

No test article-related microscopic changes were observed in the reproductive organs of males and females treated at 1000 mg/kg/day.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

No test article-related microscopic changes were observed in the reproductive organs of males and females treated at 1000 mg/kg/day.

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

The average numbers of estrous stages per 21 days for F1 females were comparable among the dosage groups.

Reproductive function: sperm measures:

no effects observed

Description (incidence and severity):

Sperm parameters in males were unaffected by test material administration.

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

All mating and fertility parameters were unaffected by T-7485 doses as high as 300 mg/kg/day in females and 1000 mg/kg/day in males.

Effect levels (P0)

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

reproductive function (oestrous cycle)

reproductive function (sperm measures)

reproductive performance

other: No male or female reproductive toxicity was observed in P generation rats following ingestion of T-7485 at up to 1000 mg/kg/day. The NOAEL is 1000 mg/kg/day.

Results: P1 (second parental generation)

General toxicity (P1)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Animals treated at 1000 mg/kg/day exhibited a significant increase in the incidence of excess salivation.

Dermal irritation (if dermal study):

not examined

Mortality:

no mortality observed

Description (incidence):

All animals survived.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

There were no abnormal body weights and body weight gains for F1 (P1) females. Body weight gains for F1 (P1) males treated at 1000 mg/kg/day were significantly reduced for the entire precohabitation period and on the day cohabitation started.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no abnormal changes in absolute or relative food consumption values.

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:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Terminal body weights and absolute weights of the seminal vesicles (with and without fluid) of F1 (P1) males were significantly reduced in the 1000 mg/kg/day dose group. Terminal body weights were significantly increased in F1 (P1) females treated at any T-7485 dose. Relative liver weights were increased in F1 (P1) males treated at 1000 mg/kg/day.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were not test article-related gross pathological findings at any dose level.

Neuropathological findings:

not examined

Description (incidence and severity):

Microscopic examination of the liver of F1 (P1) males and the kidneys of F1 (P1) males and females revealed test article-related effects similar to those in P males and females. No test article-related microscopic changes were observed in the reproductive organs of F1 (P1) males and females treated at 1000 mg/kg/day.

Histopathological findings: neoplastic:

no effects observed

Description (incidence and severity):

Microscopic examination of the liver of F1 (P1) males and the kidneys of F1 (P1) males and females revealed test article-related effects similar to those in P males and females. No test article-related microscopic changes were observed in the reproductive organs of F1 (P1) males and females treated at 1000 mg/kg/day.

Reproductive function / performance (P1)

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

The age of vaginal patency and average numbers of estrous stages per 21 days for F1 (P1) females were comparable among the dosage groups.

Reproductive function: sperm measures:

no effects observed

Description (incidence and severity):

Sperm parameters in F1 (P1) males were unaffected by test material administration.

Reproductive performance:

no effects observed

Description (incidence and severity):

Natural delivery observations were unaffected by test material administration. The number of surviving pups per litter, the percentage of male pups, litter size, and average pup body weight per litter were comparable among the dose groups.

Effect levels (P1)

Key result

Dose descriptor:

NOAEL

Effect level:

ca. 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

reproductive function (oestrous cycle)

reproductive function (sperm measures)

reproductive performance

other: No male or female reproductive toxicity was observed in F1 (P1) generation rats following ingestion of T-7485 at up to 1000 mg/kg/day. The NOAEL is 1000 mg/kg/day.

Results: F1 generation

General toxicity (F1)

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Animals treated at 1000 mg/kg/day exhibited a significant increase in the incidence of excess salivation.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

no mortality observed

Description (incidence and severity):

All animals survived.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

There were no abnormal body weights and body weight gains for F1 females. Body weight gains for F1 males treated at 1000 mg/kg/day were significantly reduced for the entire precohabitation period and on the day cohabitation started.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no abnormal changes in absolute or relative food consumption values.

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:

not examined

Urinalysis findings:

not examined

Sexual maturation:

no effects observed

Description (incidence and severity):

The age of vaginal patency and average numbers of estrous stages per 21 days for F1 females were comparable among the dosage groups.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Terminal body weights and absolute weights of the seminal vesicles (with and without fluid) of F1 males were significantly reduced in the 1000 mg/kg/day dose group. Terminal body weights were significantly increased in F1 females treated at any T-7485 dose. Relative liver weights were increased in F1 males treated at 1000 mg/kg/day.

Gross pathological findings:

no effects observed

Description (incidence and severity):

There were not test article-related gross pathological findings at any dose level.

Histopathological findings:

no effects observed

Description (incidence and severity):

Microscopic examination of the liver of F1 males and the kidneys of F1 males and females revealed test article-related effects similar to those in P males and females. No test article-related microscopic changes were observed in the reproductive organs of F1 males and females treated at 1000 mg/kg/day.

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not examined

Effect levels (F1)

Key result

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

ca. 300 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

other: Developmental toxicity at 1000 mg/kg/day cannot be evaluated due to presence of maternal and paternal toxicity. The NOAEL for developmental effects is 300 mg/kg/day.

Results: F2 generation

General toxicity (F2)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

No clinical or necropsy observations for F2 generation pups were attributable to dosages of the test substance as high as 1000 mgkg/day because: 1) the incidences were not dosage-dependent; and 2) the observation occurred in only one or two litters. Clinical observations included cold to touch, dehydration, not nesting, not nursing, lower midline, chest, head and neck, hindpaw or entire body purple, exophthalmos, corneal opacity, yellow skin, no milk in stomach, scab or abrasion on hindlimb, emaciation and tip of tail missing or black.

Dermal irritation (if dermal study):

not examined

Mortality / viability:

mortality observed, non-treatment-related

Description (incidence and severity):

No treatment-related mortality was observed.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

F2 generation weanling rats had comparable terminal $body weights on DL 22; the values did not' significantly differ.

Food consumption and compound intake (if feeding study):

not examined

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:

not examined

Urinalysis findings:

not examined

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

None of the significant differences in organ weights or organ weight ratios for the F2 generation pups were considered treatment-related because they were not dosage dependent.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

No treatment-related gross pathological findings were observed.

Histopathological findings:

not examined

Developmental neurotoxicity (F2)

Behaviour (functional findings):

not examined

Developmental immunotoxicity (F2)

Developmental immunotoxicity:

not examined

Effect levels (F2)

Key result

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

ca. 1 000 mg/kg bw/day (actual dose received)

Based on:

act. ingr.

Sex:

male/female

Basis for effect level:

viability

clinical signs

mortality

body weight and weight gain

organ weights and organ / body weight ratios

gross pathology

other: Terminal body weights did not differ among the groups. Significant differences in organ weights, organ weight ratios and viability were not considered treatment-related. The male and female NOAEL was 1000 mg/kg.

Overall reproductive toxicity

Key result

Reproductive effects observed:

no

Lowest effective dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Treatment related:

no

Any other information on results incl. tables

Based on the results of this study, no female reproductive toxicity was observed in P or F1 generation rats following ingestion of T-7485 at up to 1000 mg/kg/day. The reproductive NOAEL is 1000 mg/kg/day.

Applicant's summary and conclusion

Conclusions:

Based on the results of this study, no male or female reproductive toxicity was observed in P or F1 generation rats following ingestion of T-7485 at up to 1000 mg/kg/day. The reproductive NOAEL is 1000 mg/kg/day. Developmental toxicity at 1000 mg/kg/day cannot be evaluated due to presence of maternal and paternal toxicity. The NOAEL for developmental effects is 300 mg/kg/day.

Executive summary:

The reproductive and developmental toxicity of T-7485 was evaluated in Sprague Dawley rats in a 2-generation study. This study was performed in compliance with OECD GLP (1997) and US EPA GLP 40 CFR 160 and 792. The study design was based on US EPA OPPTS 870.3800 (1998), and US EPA TSCA Test Guideline 799.9380 (1997). The test material was prepared in 0.1% carboxymethylcellulose in deionized water (vehicle). P generation rats (30/sex/dose) received 0 (vehicle), 30, 100, 300, or 1000 mg/kg/day of T-7485 via oral gavage at a dose volume of 10 mL/kg beginning at approximately 6 weeks of age (at least 70 days before cohabitation) and continuing until the day before sacrifice (males: after completion of cohabitation period; females: DL 22). All F1 pups were weaned on DL 22 and were randomly selected for continued evaluation. F1 pups (30/sex/dose) received 0 (vehicle), 30, 100, 300, or 1000 mg/kg/day of T-7485 via oral gavage beginning at weaning (approximately 70 days before cohabitation) and continuing until the day before sacrifice (males: after completion of cohabitation period; females: DL 22). 

Parameters evaluated in P and F1 generations: clinical observations (daily), body weights and food consumption (males: weekly; females: weekly; DGs 0, 7, 10, 14, 18, 21, and 25; and DLs 1, 5, 8, 11, 15, and 22), estrous cycling, duration of gestation, fertility index, gestation index, number and sex of offspring per litter, number of implantation sites, general condition of dam and litter during postpartum period, litter size and viability, viability indices, lactation index, percent survival, sex ratio, maternal behavior, gross necropsy, sperm count and motility, organ weights of select organs, and histopathology of select organs and tissues. F1 females and males were also evaluated for age of vaginal patency and age of preputial separation, respectively. Parameters evaluated in F2 generation: number and sex of pups, stillbirths, live births, pup body weights, and gross external alterations.

P generation: One male treated at 1000 mg/kg/day was found dead on Day 55 as a result of an intubation accident. All other P generation animals survived until scheduled termination. Males treated at 1000 mg/kg/day exhibited significant increases in the incidences of excess salivation, perioral substance, chromorhinorrea, and urine-stained abdominal fur. One or two males treated at 300 mg/kg/day exhibited excess salivation and perioral substance that were considered treatment-related. Females treated at 1000 mg/kg/day exhibited a significant increase in the incidence of dried or wet red perioral substance and excess salivation during precohabitation and gestation. At 1000 mg/kg/day, male body weight gains were significantly reduced on Days 43 to 50 and 50 to 57, and significant body weight loss occurred on Days 105 to 112. Body weights and body weight gains of females during precohabitation, gestation, and lactation were unaffected by T-7485 doses as high as 300 mg/kg/day. Terminal body weights of males and females were unaffected by test article administration. There were no abnormal changes in absolute or relative food consumption values. All mating and fertility parameters were unaffected by T-7485 doses as high as 300 mg/kg/day in females and 1000 mg/kg/day in males. The average numbers of estrous stages per 21 days for F1 females were comparable among the dosage groups. There were no test article-related gross necropsy findings in males or females. Absolute and relative liver weights were significantly increased in males treated at 300 or 1000 mg/kg/day. Absolute brain weight was significantly reduced in 1000 mg/kg/day-treated females. Treatment-related microscopic changes were observed in the liver of males treated at 300 or 1000 mg/kg/day and in the kidneys of males and females treated at 300 or 1000 mg/kg/day. No test article-related microscopic changes were observed in the reproductive organs of males and females treated at 1000 mg/kg/day. Sperm parameters in males were unaffected by test material administration. Natural delivery observations were unaffected by test article administration. Terminal body weights of F1 pups were comparable among the dose groups. There were no test article-related clinical or necropsy observations in F1 pups at any dose. 

F1 generation: All F1 animals survived. Animals treated at 1000 mg/kg/day exhibited a significant increase in the incidence of excess salivation. There were no abnormal body weights and body weight gains for F1 females. Body weight gains for F1 males treated at 1000 mg/kg/day were significantly reduced for the entire precohabitation period and on the day cohabitation started. There were no abnormal changes in absolute or relative food consumption values. Preputial separation of F1males was slightly delayed (approximately 2 days later than control group value) in the 1000 mg/kg/day dose group. This observation was considered to be associated with the significantly reduced body weights at the time of sexual maturation. The age of vaginal patency and average numbers of estrous stages per 21 days for F1 females were comparable among the dosage groups. All female and male mating and fertility parameters were unaffected by test article administration. All necropsy observations were considered unrelated to test article administration. Terminal body weights and absolute weights of the seminal vesicles (with and without fluid) of F1 males were significantly reduced in the 1000 mg/kg/day dose group. Terminal body weights were significantly increased in F1 females treated at any T-7485 dose. Relative liver weights were increased in F1 males treated at 1000 mg/kg/day. Microscopic examination of the liver of F1 males and the kidneys of F1 males and females revealed test article-related effects similar to those in P males and females. No test article-related microscopic changes were observed in the reproductive organs of F1 males and females treated at 1000 mg/kg/day. Sperm parameters in F1 males were unaffected by test material administration. Natural delivery observations were unaffected by test material administration. The number of surviving pups per litter, the percentage of male pups, litter size, and average pup body weight per litter were comparable among the dose groups. There were no test article-related clinical or necropsy observations in F2 pups at any dose. No test article-related differences in F2 pup absolute and relative organ weights or terminal body weights among the dose groups.

Based on the results of this study, no male or female reproductive toxicity was observed in P or F1 generation rats following ingestion of T-7485 at up to 1000 mg/kg/day. The reproductive NOAEL is 1000 mg/kg/day. Developmental toxicity at 1000 mg/kg/day cannot be evaluated due to presence of maternal and paternal toxicity. The NOAEL for developmental effects is 300 mg/kg/day.