Registration Dossier

Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
reproductive toxicity, other
Remarks:
Dose Range finder for the extended one-generation reproductive performance study
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
From 2020-06-17 to 2020-08-29 (day of necropsy)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000

Materials and methods

Test guideline
Qualifier:
no guideline required
Principles of method if other than guideline:
Due to the preliminary nature of this study no specific regulations or guidelines are applicable.
GLP compliance:
no
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium trifluoroacetate
EC Number:
220-879-6
EC Name:
Sodium trifluoroacetate
Cas Number:
2923-18-4
Molecular formula:
C2HF3O2.Na
IUPAC Name:
sodium trifluoroacetate
Test material form:
solid
Details on test material:
- Batch number: 2019052304
- Appearance: White solid
- Storage conditions: At ambient temperature (15 to 25°C)
- Expiry date: 01 May 2021
- Purity: 99.9%
- Impurity: water, 0.0635%
Specific details on test material used for the study:
Trifluoroacetic acid (TFA) is a corrosive liquid. To identify the systemic hazards of the substance, testing with the neutral salt sodium trifluoroacetate is considered appropriate to avoid local corrosive effects at the site of administration.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS Limited, UK
- Strain: Wistar rat, RccHan™;WIST
- Sex: females; stock males used for mating
- Age of F0 animnals at the start of the study (GD0): 77 to 83 days old
- Weight range of F0 animals at the start of the study (GD0): 174 to 204 g
- Housing:
♦ During acclimatisation, up to 4 animals per sex per cage in solid bottomed polycarbonate cages, with bedding (softwood based bark-free fiber).
♦ During mating, 1 male/1 female in polycarbonate cages with a stainless steel mes lid and floor.
♦ After mating, during gestation, birth and lactation, the females were individually housed in solid bottomed polycarbonate cages, with bedding (softwood based bark-free fiber).
♦ The selected F1 animals were pair housed as siblings in solid bottomed polycarbonate cages, with bedding (softwood based bark-free fiber).
- Diet: SDS VRF1 Certified, powdered diet, ad libitum.
- Water: Potable water from the public supply via polycarbonate bottles with sipper tubes. Bottles were changed at appropriate intervals, ad libitum.
- Acclimation period: 5 days before commencement of pairing

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Monitored and maintained within the range of 20-24ºC.
- Humidity (%): Monitored and maintained within the range of 40-70%.
- Air changes: not reported, Filtered fresh air which was passed to atmosphere and not recirculated
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 24 June 2020 To: 29 August 2020

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Before commencement of treatment, the suitability of the proposed mixing procedures was determined and stability and homogeneity at a concentration of 50 to 10000 ppm was determined as part of another study (Covance Study no. LK56VW).

DIET PREPARATION
- Rate of preparation of diet (frequency): Once weekly
- Mixing appropriate amounts with (Type of food): The test substance was mixed with SDS VRF1 Certified, powdered diet. The test substance was incorporated into the diet to provide the required concentrations by initial preparation of a premix. The amount of test substance required for the premix was added to an equal amount of plain diet and stirred. An amount of plain diet equal to the weight of the mixture was added and the mixture was stirred again until visibly homogenous. The doubling up process was repeated until approximately half the premix diet was added. At this stage the mixture was ground with a mechanical grinder. The mixture was made up to the weight of the premix with plain diet. The premix was then mixed using a turbula mixer for 200 cycles. This premix was diluted with further quantities of plain diet using the doubling up process to prepare the test mixes. Each formulation was mixed using a Turbula mixer for 200 cycles.
- Storage temperature of food: Ambient (15 to 25°C). A stability of 22 days under these conditions was confirmed.
Details on mating procedure:
- M/F ratio per cage: 1/1
- Duration of pairing: Nominally four days
- Proof of pregnancy: vaginal plug/sperm in vaginal smear referred to as Day 0 of pregnancy (GD 0)
- After successful mating each pregnant female was caged: individually
Analytical verification of doses or concentrations:
no
Details on analytical verification of doses or concentrations:
No formulation analysis was performed in this study.
Duration of treatment / exposure:
- F0 females: From Day 6 after mating up to weaning of the F1 offspring.
- F1 animals: From mid lactation (when offspring would be expected to start to consume diet) until Day 35 of age.
During the lactation phase, from Day 1 to Day 21, females received reduced dietary concentrations of 700, 1700 or 4200 ppm. Due to the preliminary nature of this study no specific regulations or guidelines are applicable.
Frequency of treatment:
Continuously via the diet
Doses / concentrationsopen allclose all
Dose / conc.:
1 400 ppm (nominal)
Remarks:
Main phase
Dose / conc.:
3 400 ppm (nominal)
Remarks:
Main phase
Dose / conc.:
8 400 ppm (nominal)
Remarks:
Main phase
Dose / conc.:
700 ppm (nominal)
Remarks:
Lactation phase
Dose / conc.:
1 700 ppm (nominal)
Remarks:
Lactation phase
Dose / conc.:
4 200 ppm (nominal)
Remarks:
Lactation phase
No. of animals per sex per dose:
F0 generation: 6 females per group
F1 generation: 6 males and 6 females per group. The offspring with the lowest within-litter identification per sex from each selected litter was selected to form the F1 generation, after exclusion of grossly atypical animals (where possible, one male and one female from each litter). Selected animals were microchipped on Day 18-20 of age and separated from littermates on Day 21 of age.
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: The dose levels of 1400, 3400, and 8400 ppm, corresponding to approximately 100, 250 and 625 mg/kg/day, were selected based on the results of a combined Repeated Dose Toxicity Study with Reproduction/Developmental Toxicity Screening (OECD TG422, CitoxLab 2012) supported by a 90-day repeat dose study in rats (Bayer 2007, Report no: SA 06080).

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Animals were inspected visually at least twice daily for evidence of ill-health or reaction to treatment.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: A detailed physical examination was performed on each animal to monitor general health at the following time points:
♦ F0 females: Gestation days 0, 5, 12, 18 and 20. Lactation days 1, 7, 14 and 21.
♦ Selected F1 generation: once each week from from PND 21 to PND 35.

BODY WEIGHT: Yes
- Time schedule for examinations: F0 Females: Gestation days 0, 3, 6, 10, 14, 17 and 20 and Lactation days 1, 4, 7, 11, 14, 18 and 21. F1 selected animals: Twice weekly from PND 21 to PND 35.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Food consumption was measured as follows:
♦ F0 females: Gestation days 0-2, 3-5, 6-9, 10-13, 14-16 and 17-19. Lactation days 1-3, 4-6, 7-10, 11-13, 14-17 and 18-20.
♦ Selected F1 generation: Twice weekly from PND 21 to PND 35.
From these records the mean daily consumption per animal (g/animal/day) was calculated for each phase.

OTHER:
- Parturition observations: From Day 20 after mating, females were inspected three times daily for evidence of parturition. The progress and completion of parturition was monitored, numbers of live and dead offspring were recorded and any difficulties observed were recorded.
Oestrous cyclicity (parental animals):
Not examined.
Sperm parameters (parental animals):
Not examined.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: yes
- If yes, maximum of 8 pups/litter (4/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in F1 offspring during littering phase:
- Clinical observations: Examined at approximately 24 hours after birth (Day 1 of age) and then daily thereafter for evidence of ill health or reaction to treatmentd
- Litter size: Daily records were maintained of mortality and consequent changes in litter size from Days 1 to 21 of age.
- Sex ratio of each litter: Recorded on Days 1, 4 (before and after culling) and 11, 14, 17 and 21 of lactation
- Individual offspring body body weights: Days 1, 4, 7, 11, 14, 17 and 21 of age.

GROSS EXAMINATION OF DEAD PUPS:
yes, where possible, a fresh macroscopic examination with an assessment of stomach for milk content was performed. Abnormal tissues were retained in an appropriate fixative.
Postmortem examinations (parental animals):
SACRIFICE
- Maternal animals: Surviving F0-females were sacrificed on LD21. F0-females that failed to produce viable litters were sacrificed on day 25 after mating. Mothers of litters that dyed before weaning were killed on day on which the last offspring died.

GROSS NECROPSY
- After a review of the history of each animal, a full macroscopic examination of the tissues was performed. All external features and orifices were examined visually. Any abnormality in the appearance or size of any organ and tissue (external and cut surface) was recorded and the required tissue samples preserved in appropriate fixative.

HISTOPATHOLOGY / ORGAN WEIGHTS
- The liver was weighed and samples fixed for all F0 females. These samples were not processed histologically, but retained against any future requirement for microscopic examination. For F0-females the number of implantation sites was recorded. In addition, for females whose litters died before LD 21 the appearance of mammary tissue was checked.
Postmortem examinations (offspring):
SACRIFICE
- Unselected F1 offspring were sacrificed at culling on PND 4, at scheduled kill on LD 21, or selected spares after establishment of selected F1 generation.
- Selected F1 animals were sacrificed on Day 35 of age

GROSS NECROPSY
- Culled offspring with no clinical signs on Day 4 of age were killed and discarded without necropsy examination. Unselected offspring at scheduled kill on Day 21 of age and selected offspring on Day 35 were subject to complete macroscopic examination. Abnormal tissues retained in an appropriate fixative.

HISTOPATHOLOGY / ORGAN WEIGTHS
The liver was weighed and samples fixed for all F0 females. These samples were not processed histologically, but retained against any future requirement for microscopic examination. The number of implantation sites were recorded for each females.
Statistics:
The following sequence of statistical tests was used for body weight, food consumption, implantations, litter size, sex ratio, post implantation survival index and organ weight data:
A parametric analysis was performed if Bartlett's test for variance homogeneity (Bartlett, 1937) was not significant at the 1% level. For pre-treatment data, analysis of variance was used to test for any group differences. Where this was significant (p<0.05) inter group comparisons using t-tests, with the error mean square from the one-way analysis of variance, were made. For all other analyses the F1 approximate test was applied.
If the F1 approximate test for monotonicity of dose-response was not significant at the 1% level, Williams' test for a monotonic trend was applied. If the F1 approximate test was significant, suggesting that the dose response was not monotone, Dunnett's test (Dunnett 1955, 1964) was performed instead.
A non-parametric analysis was performed if Bartlett's test was still significant at the 1% level following both logarithmic and square-root transformations. For pre-treatment data, Kruskal-Wallis’ test (Kruskal and Wallis, 1952; 1953) was used to test for group differences. Where this was significant (p<0.05) inter group comparisons using Wilcoxon rank sum tests were made. For all other analyses the H1 approximate test, the non parametric equivalent of the F1 test, was applied.
- For live birth and viability indices dichotomized to 1 when 100% and 0 otherwise, if the Cochran Armitage test was significant at the 5% level, then the direction of the trend was established and one-tailed step-down testing in this direction was performed. If the Cochran-Armitage test was not significant at the 5% level, then a Chi-square test was applied. If the Chi-square test was significant at the 5% level, the treatment groups were compared using pairwise comparisons of each dose group against the Control using Fisher’s exact tests; otherwise, no further comparisons were made.
Reproductive indices:
♦ Gestation index (%) = (Number of live litters born) / (Number of pregnant) x 100
Offspring viability indices:
♦ Post-implantation survival index (%) = (Total number of offspring born) / (Total number of uterine implantation sites) x 100
♦ Live birth index (%) = (Number of live offspring on Day 1 after littering) / (Total number of offspring born) x 100
♦ Viability index (%) = (Number of live offspring on Day 4 before culling) / (Number live offspring on Day 1 after littering) x 100
♦ Lactation index (%) = (Number of live offspring on Day 21 after littering) / (Number of live offspring on Day 4 (after culling) x 100
♦ Percentage males = (Number of males in litter) / (Total number of offspring in litter) x 100. The percentage of male offspring in each litter was calculated at Day 1, and for live offspring on Days 1, 4 (before and after culling) 11, 14, 17 and 21 of age.

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
There were no signs at routine physical examination that could be attributed to administration of Sodium Trifluoroacetate.
Dermal irritation (if dermal study):
not examined
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Following the start of treatment on GD6 animals receiving Sodium Trifluoroacetate showed low body weight gain up to GD10 (p<0.01); a dose response was apparent. At 8400 ppm the overall body weight gain for females from GD6 to GD20 was marginally low at approximately 92% of Controls although this difference did not attain statistical significance; body weight gain at 1400 or 3400 ppm over the same period was similar to Controls.
Absolute body weights for all treated females were low when compared with Controls during lactation with the difference at 4200 (8400) ppm attaining statistical significance (p<0.05) on Lactation Day 1 (LD1); a dose response was not apparent. However, overall body weight gains for treated females showed no adverse effect of treatment. Detailed results are provided in Table 1 (see attached document).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At 8400 ppm food consumption was low from GD6 to GD19 at approximately 88% of Controls; with the mean value for GD6-9 attaining statistical significance (p<0.05). Food consumption at 1400 or 3400 ppm over the same period was similar to Controls. During lactation food consumption for females receiving 4200 (8400) ppm was low at approximately 87% Controls, with the difference attaining statistical significance (p<0.05) on LD4 to LD6; food consumption at 700 (1400) or 1700 (3400) ppm was similar to Controls.
Achieved dose levels in F0 females are provided in Table 2 (see attached document).
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
Endocrine 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:
not examined
Histopathological findings: neoplastic:
not examined
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not examined
Reproductive function: sperm measures:
not examined
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
- Gestation index: The duration of gestation and the gestation index were unaffected by administration of Sodium Trifluoroacetate at dose levels up to and including 8400 ppm
- Litter size/Survival indices: Live birth index was slightly low at 4200 (8400) ppm, with viability index on Day 4 unaffected, resulting in a marginally low mean litter size prior to litter standardisation on Day 4 of age. Offspring survival from Day 4 of age was unaffected by administration of the test item. Litter size and survival at 700 (1400) or 1700 (3400) ppm showed no differences that could be related to treatment.
- Sex ratio: There was no effect of maternal treatment on sex ratio at dose levels up to and including 8400/4200ppm.

Results: F1 generation

General toxicity (F1)

Clinical signs:
no effects observed
Description (incidence and severity):
The clinical condition of offspring was unaffected by administration of Sodium Trifluoroacetate to maternal animals at dose levels up to and including 4200 (8400) ppm.
Dermal irritation (if dermal study):
not examined
Mortality / viability:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Unselected offspring (LD1-21): On Day 1 of age the mean absolute body weight for both male and female offspring were low when compared with Controls (p<0.05) at 4200 (8400) ppm and marginally low at 1700 (3400) ppm. Body weight gain up to Day 4 of age was essentially similar across the groups, however during Days 4-7 of age weight gain for offspring at all dose levels was significantly low when compared with Controls (p<0.05); a dose response was apparent for males but not females. Thereafter, body weight gains remained marginally low at 4200 (8400) ppm. Overall body weight gain (LD1-21) for offspring at 4200 (8400) ppm was low when compared with Controls (p<0.05; approximately 87% of Controls); at 700 (1400) or 1700 (3400) ppm overall body weight gain was similar to Controls.
Selected offspring (Day 21-35): At weaning on Day 21 of age mean bodyweight for males and females at 8400 ppm was low at approximately 89% of Controls (p<0.05); mean bodyweights at 1400 or 3400 ppm were similar to Controls. Subsequent body weight gain from Day 21 to Day 24 of age was similar to Controls for both male and females at all dietary concentrations. From Day 24 of age selected male animals showed low body weight gain when compared with Controls and the overall weight gain at 8400 ppm from Day 21 to Day 35 of age was approximately 79% of Controls; males receiving 1400 or 3400 ppm showed weight gain that was similar to Controls. From Day 24 to Day 28 female animals at 3400 or 8400 ppm showed slightly but significantly low body weight gain when compared with Controls (p<0.05). The overall body weight gain for selected females from Day 21 to Day 35 of age was low at approximately 85% of Controls for females at 3400 ppm however this difference was not apparent at 1400 or 8400 ppm. Detailed results are provided in Table 4 (see attached document).
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Overall food consumption from Day 21 to Day 34 of age was marginally low for animals at 3400 or 8400 ppm at approximately 89/90% of Controls; food consumption at 1400 ppm was similar to Controls. Achieved dose levels in F1 animals are provided in Table 5 (see attached document).
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
Anogenital distance (AGD):
not examined
Nipple retention in male pups:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The mean body weight relative liver weight for both male and female animals were high when compared with Controls (p<0.01) at all dietary concentrations; a dose response was apparent. Detailed results are provided in Table 6 (see attached document).
Gross pathological findings:
no effects observed
Description (incidence and severity):
Macroscopic examination of offspring that died prematurely and those at scheduled termination on Day 21 of age did not reveal any findings that could be attributed to Sodium Trifluoroacetate. Macroscopic examination of selected F1 animals on Day 35 of age did not reveal any findings that could be attributed to administration of Sodium Trifluoroacetate.
Histopathological findings:
not examined
Other effects:
not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not examined

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not examined

Overall reproductive toxicity

Reproductive effects observed:
no

Any other information on results incl. tables

Proof of absorption

Blood samples were collected 0.5-1hr, 4.5-5hr and 8.5-9hr after light was switched on. Blood was collected during the gestation period (GD 17) and the lactation period (LD11) from n=6/females per dose group. The first data point at 0.5-1hr was also used as a 24-h value in the TK calculations[1].


TFA groups:

F0: At GD 17 mean maximum concentrations of 214, 307 and 485 mg/L were measured at the 0.5-1 hour sampling time point on GD 17. AUC242values were 4819, 6892 and 10285 µg*h/mL for the low, mid-and high-dose group. Compared to the external applied / achieved dose level the internal dose (described as AUC24) showed a sub-proportional increase, indicating that the absorption is affected.

Comparable results were found for the measurements on LD 11. Mean maximum concentrations were 171 mg/L at 4.5-5 hours at the low dose and 279 and 405 mg/L at 0.5-1 h in the mid- and high-dose group. AUC24 values were 4032, 6334 and 9304 µg*h/mL.

 

F1: For F1 animals on PND 4 and 21 only one sample was taken, therefore it is not possible to calculate an AUC24.

In pooled samples from PND 4 males and females TFA plasma concentrations were much lower without showing a dose-correlation. Mean cMaxvalues on PND 4 (measured for pooled samples from males and females) were comparable in all dose groups, with values of 36.4, 33.7 and 34.2 mg/L at the low, mid- and high dose. At PND 21 (unclear when plasma samples were taken) TFA plasma concentrations in F1 weanlings were similar to the values of the first sampling timepoint at 0.5-1 hr in dams. Mean plasma concentrations were 140, 289 and 342 mg/L in males, and 142, 276 and 394 mg/L in females at the low- mid- and high-dose, respectively.

 

In dams, relatively high initial plasma concentrations were noted at the 0.5-1 hr sampling time point across all dose groups which did not decline relevantly within the sampling interval of ≈ 9 hours, assuming either a strong oral absorption and/or a slow clearance from blood (probably saturation of excretion/metabolism). Increasing the external dose from 97.5 to 230 mg/kg bw was associated with a slightly less than dose-proportional increase in the cMaxand the AUC24. Internal exposure still increased from ≈ 230 to 547 mg/kg bw, but in a clearly less-than dose proportional manner.

[1] The AUC24 is calculated, as described by Jochemsen et. al. 1993, by taking the concentrations measured during the daylight hours and using the concentration measured at lights on again at 24 hours post the original time point to cover night-time exposure.

Jochemsen R, Bazot D, Brillanceau MH and Lupart M (1993). Assessment of drug exposure in rat dietary studies. Xenobiotica, 23(10), 1145-1154

Applicant's summary and conclusion

Conclusions:
On the basis of this preliminary reproductive study in rats, dose levels of 10, 50 and 250 mg/kg bw/day were selected for the subsequent Extended One-generation Reproductive Study.
Executive summary:

This study was performed to assess the influence of sodium trifluoroacetate on reproductive performance when administered continuously via the diet to female Han Wistar rats, and to establish suitable treatment levels for the Extendend one-generation reproductive performance study. In the F0 generation, three groups of six female rats received Sodium Trifluoracetate at dietary concentrations of 1400, 3400 or 8400 ppm orally, during gestation from Day 6 after mating. During the lactation phase, from Day 1 to Day 21, females received reduced dietary concentrations of 700, 1700 or 4200 ppm. A similarly constituted Control group received untreated basal diet. In the F1 generation, six males and six females were treated from weaning to their scheduled termination on Day 35 of age at the same dietary concentrations as the F0 generation main phase. A similarly constituted Control group received untreated basal diet.

During the study, for the F0 generation, clinical condition, body weight, food consumption, gestation length and parturition observations, organ weight and macroscopic pathology investigations were undertaken. For the F1 generation, clinical condition, body weight, food consumption, organ weight and macroscopic pathology investigations were undertaken. The clinical condition, litter size and survival, sex ratio and body weight for all offspring were also assessed. In addition, to determine systemic exposure, plasma concentrations of TFA were determined at each dosage level.

Results

F0 Maternal responses

During gestation females receiving 1400, 3400 or 8400 ppm the mean achieved dose levels were 97.5, 230 or 547 mg/kg/day and during the first two weeks of lactation the achieved dose levels for females receiving 700, 1700 or 4200 ppm were 110.2, 262 or 578 mg/kg/day. Clinical condition, gestation length, gestation index and macropathology of F0 females was unaffected by administration of Sodium Trifluoroacetate. At 8400 ppm the overall body weight gain and food consumption for females from GD6 to GD20 was low at approximately 92% and 88% of Controls respectively; body weight gain and food consumption at 1400 or 3400 ppm over the same period was similar to Controls. During lactation the absolute body weights for all treated females were low when compared with Controls; however, overall body weight gains for treated females showed no adverse effect of treatment.  During lactation food consumption for females receiving 4200(8400) ppm was low at approximately 87% Controls; food consumption at 700(1400) or 1700(3400) ppm was similar to Controls. On Day 21 of lactation the mean body weight relative liver weights for females that received 1700(3400) or 4200(8400) ppm were slightly high at approximately 111% of Controls; however, these differences did not attain statistical significance. Liver weight at 700(1400) ppm was similar to Controls.

F1 Litter Responses

General condition and macropathology for offspring were unaffected by administration of Sodium Trifluoroacetate to maternal animals at dose levels up to and including 4200 (8400) ppm. Live birth index was slightly low at 4200 (8400) ppm, with viability index on Day 4 unaffected, resulting in a marginally low mean litter size prior to litter standardisation on Day 4 of age. Offspring survival from Day 4 of age was unaffected by administration of the test item.

Litter size and survival at 700(1400) or 1700(3400) ppm showed no differences that could be related to treatment and there was no effect of maternal treatment on sex ratio at dose levels up to and including 8400/4200ppm.

On Day 1 of age the mean absolute body weight for both male and female offspring were low when compared with Controls (p<0.05) at 4200(8400) ppm and marginally low at 1700(3400) ppm. Overall body weight gain (LD1-21) for offspring at 4200(8400) ppm was low when compared with Controls (p<0.05; approximately 87% of Controls); at 700(1400) or 1700(3400) ppm overall body weight gain was similar to Controls.

F1 Generation Responses

The mean achieved dose levels for selected animals from weaning up to Day 34 of age was 202, 476 and 1290 mg/kg/day for males and 228, 557 and 1369 mg/kg/day for females at 1400, 3400 and 8400 ppm, respectively. Clinical condition and macropathology of F1 females were unaffected by administration of Sodium Trifluoroacetate. At weaning on Day 21 of lactation mean bodyweight for males and females at 8400 ppm was low at approximately 89% of Controls (p<0.05); mean bodyweights at 1400 or 3400 ppm were similar to Controls. At 8400 ppm the overall weight gain from Day 21 to Day 35 of age was approximately 79% of Controls for males and 85% of Controls for females; overall weight gain for animals receiving 1400 or 3400 ppm was similar to Controls.

Overall food consumption from Day 21 to Day 34 of age was marginally low for animals at 3400 or 8400 ppm at approximately 89/90% of Controls; food consumption at 1400 ppm was similar to Controls.

The mean body weight relative liver weight for both male and female animals were high when compared with Controls (p<0.01) at all dietary concentrations; a dose response was apparent.

In dams, relatively high initial plasma concentrations were noted at the 0.5-1 hr sampling time point across all dose groups which did not decline relevantly within the sampling interval of ≈ 9 hours, assuming either a strong oral absorption and/or a slow clearance from blood (probably saturation of excretion/metabolism). Increasing the external dose from 97.5 to 230 mg/kg bw was associated with a slightly less than dose-proportional increase in the cMaxand the AUC24. Internal exposure still increased from ≈ 230 to 547 mg/kg bw, but in a clearly less-than dose proportional manner.