Potassium trifluoroacetate

Endpoint summary

Description of key information

Oral exposure to the reaction mass of TFSK/TFAK (OECD 422, GLP) did not result in any treatment related findings and the No Observed Adverse Effect Level (NOAEL) for oral subacute toxicity was considered to be 1000 mg/kg/day of active ingredient.

Oral exposure to the reaction mass of TFSK/TFAK (OECD 408, GLP) resulted in increased relative and absolute liver weights in male and female animals and treatment-related changes in blood biochemistry parameters (ALP and T-Bil). The NOAEL for subchronic toxicity was considered to be 100 and 300 mg/kg/day of active ingredient for males and females, respectively.

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 23 May 2018 to 11 November 2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

For the Reaction mass of potassium trifluoroacetate and potassium trifluoromethanesulphinate a new chemical substance notification in China has been prepared under the Regulation ‘Measures on the Environmental Management of New Chemical Substances' (Decree No. 7 of the Ministry of Environmental Protection of the P.R. China, also known as ‘China REACH’). Under this regulation a 90-day toxicity study according to OECD 408 is part of the data requirements for substances that are produced or imported in volumes >100 t/y. For this reason an OECD 408 study was performed in 2020 at Shenyang Research Institute of Chemical Industry. The results of the study are included in the dossier and serve as the valid test related to this endpoint.

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

GLP compliance:

yes (incl. QA statement)

Remarks:

2019-02-11

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: SPF (Beijing) Biotechnology Co., Ltd, China
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 6 to 7 weeks on arrival
- Weight at study initiation: 201 to 240 g (males) and 171 to 203 g (females). At the grouping of the experiment, the weight variation in the same group used was ± 20% of the mean body weight for either sex.
- Fasting period before study: no
- Assigned to test groups randomly: yes
- Housing: animals were housed in suspended, stainless steel cages (L32.0xW28.0xH20.0cm). Animals were housed in groups of max. 2.
- Diet (e.g. ad libitum): SPF rodent growth and breeding feed supplied by Shenyang Mao Hua biological science and Technology Co., Ltd, ad libitum
- Water (e.g. ad libitum): Purified drinking water, ad libitum
- Acclimation period: 6 days. Clinical observations were performed daily until grouping.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25°C
- Humidity (%): 40-70
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: May 23, 2019 (animal arrival) To: August 29, 2019 (final sacrifice)

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Dosing solutions of the test item in the vehicle (drinking water) were prepared daily before the administration and were used within 4 hours after preparation. The test item concentrations were based on the active ingredient of the reaction mass (Potassium triflinate, TFSK: 14.4%, Potassium trifluoroacetate, TFAK: 14.1%). A dosing volume of 10 mL/kg was applied for all animals, which was adjusted based on the latest body weight.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

CONCENTRATION VERIFICATION: Before the initiation of the main toxicity study, the stability of test item solutions was determined by a validated analytical method (Study No. G1999B0040). The formulations were found to be stable for at least 4h at room temperature (15-25°C). The concentration of the formulations was determined by analysis on the first dosing day, the 7th week and the 13th week of the dosing. Formulation samples of 100 µl were collected for analysis, at least three parallel samples each time. 100 µl of the vehicle was collected for analysis, at least two parallel samples each time. Analysis was performed with Ion Chromatography (IC) with a validated analytical method for the test item. To be considered acceptable, the actual results for the analysis of the dosing formulations were to be within ±10% of the nominal concentration.

Duration of treatment / exposure:

90 days

Frequency of treatment:

Once daily

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Active ingredient

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Active ingredient

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Active ingredient

No. of animals per sex per dose:

- 10/sex/group for low and mid dose
- 16/sex/group for high dose and control

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The study doses selected were based on the pre-study results (Study No. G1857B0001A). The pre-study results indicated that the body weight gains were not affected by the test item at the dose of 1000 mg/kg a.i. during the 22-day dosing period. There were no deaths and the animals showed normal during the 22-day dosing period. Based on these results the dose level of 1000 mg/kg was choosen as the highest dose level in the main study.
- Rationale for animal assignment: Healthy animals were used for randomization grouping based on the body weight by the Provantis 9.4.3 Animal Arrangement Module. Animals were allocated to 4 groups: 16 animals of each sex in the control group and high-dose group, 10 animals of each sex in the other dose groups. At the grouping of the experiment, the weight variation in the same group used must not exceed ±20% of the mean body weight for either sex.
- Recovery group: The reversibility of effects was assessed after a recovery period of 28-days without exposure (6/sex/group for control and high dose).

Positive control:

Not included

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: All animals were inspected for signs of morbidity and mortality at the beginning and the end of work at least once daily.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: General clinical signs were recorded at least once daily at about 1.5h after dosing for all animals during the study. During the recovery period, the observation time was performed at the same time each day. The healthy conditions and toxicity signs wase recorded.
Detailed physical examinations were performed for all animals prior to the first exposure (at grouping) and once a week during the treatment period and during the recovery period. The animals were taken out of the home cage for observation, and all the findings will be recorded in detail. Observation includes changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, piloerection, pupil size, unusual respiratory pattern), changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypes (e.g. excessive grooming, repetitive circling) or bizarre behaviors (e.g. self-mutilation, walking backwards).

BODY WEIGHT: Yes
- Time schedule for examinations: Animals were weighed at grouping, once a week during the treatment period and recovery period, and in a moribund state or at death. Animals were fasted overnight (16-18h) prior to necropsy, and empty-stomach body weights were collected before necropsy.

FOOD CONSUMPTION: yes
- Time schedule for examinations: The weight of food supplied and of that remaining at the end of the food consumption period was recorded weekly for all animals during the treatment period. From these records the mean daily consumption was calculated.

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Ophthalmological examinations were conducted in all animals after grouping and the day before the end of treatment.

CLINICAL PATHOLOGY INVESTIGATIONS: Yes
- Time schedule for collection of blood: All surviving animals at termination of dosing and after the recovery period were anesthetized by CO2 inhalation, and blood was collected via abdominal aorta for hematology, coagulation and serum biochemistry.
- Animals fasted: yes, all animals were fasted overnight before blood collection.
- Anaesthetic used for blood collection: Yes, CO2 anaesthesia
- Haematology: Parameters checked in Table 1 (see 'Any other information on materials methods') were examined.
- Coagulation: Prothrombin time, activated partial thromboplastin time and fibrinogen were investigated.
- Clinical chemistry: Parameters checked in Table 2 (see 'Any other information on materials methods') were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: Before necropsy after termination of dosing and the recovery period, urine samples were collected from all surviving animals by abdominal extrusion.
- Metabolism cages used for collection of urine: No
- Animals fasted: Animals were fasted overnight prior to necropsy, but water was available.
- The following measurements were performed on urine: Appearance (visual observation), pH, urine specific gravity, occult blood, ketones, glucose, total protein, bilirubin, urobilinogen, nitrites, leukocytes, vitamin

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: week 11 of treatment
- Dose groups that were examined: all surviving animals in control and high dose group were assessed for general behavior, sensory reactivity by different types of stimulation, grip strength and motor activity.
- Battery of functions tested:
+ General behaviour assessment: Responses to the following manipulations was assessed in the open-field observations, home-cage observations, handling and manipulative observations. The following parameters were checked: Spontaneous Activity, Gait and Posture, Reactivity and Alertness, Abnormal Motor Movements (involuntary motor movements/abnormal/stereotypic behavior), Observations of Autonomic Nerve (Lacrimation/Salivation/Piloerection/Palpebral closure/Eye abnormal/Urination/Defecation/ Breath/Muscle tension), Reflexes (Pupil/Palpebral/Pinna/Extensor Thrust Reflex)
+ Motor activity Rat: Activity Record Instrument was used to assess motor activity. The evaluation period was three minutes for each animal.
+ Forelimb and Hindlimb Grip Strength: A tensile strength tester was used. Each animal was allowed to grip the board of the meter with its forepaws and hindpaws. The animal was pulled by the base of the tail until its grip was broken. A record of the force required to break the grip for each animal was made. Three consecutive trials were performed for each animal and the average readings were used.
+ Sensory reactivity: Each animal was individually assessed for sensory reactivity to visual, auditory, and proprioceptive stimuli. Then the following parameters were observed: approach response, click response, tail pinch, touch response.

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
The necropsy included careful visual examinations of the external features of the carcass, external body orifices, the abdominal, thoracic, and cranial cavities and their contents. The location, size, hardness and the color of the abnormal findings were recorded. The following organ weights were recorded from all animals on the scheduled day of necropsy: brain, thyroid, heart, lung, liver, spleen, adrenal glands, kidney, thymus, uterus (female), testis (male), epididymis (male), prostate (male)

HISTOPATHOLOGY: Yes.
The samples of the tissues indicated in Table 3 (see 'Any other information on materials methods') were collected and preserved. The tissues from each animal were preserved in 10% neutral-buffered formalin (eyes were preserved in Davidson’s solution, and testis in Improved Davidson’s solution). Lungs were infused with fixative prior to immersion in 10% neutral-buffered formalin. Stomach, intestine and bladder were infused with formalin prior to immersion in 10% neutral-buffered formalin. Tissues and organs collected from the control and high dose group of animals and gross lesions found in the other dose group animals were sectioned, fixed and dehydrated with alcohol, embedded in paraffin, sliced using microtomy, and stained with hematoxylin and eosin, extracted/washed with dimethylbenzene, sealed with rosin and examined microscopically.
Histopathology was examined on the preserved organs and tissues of all animals in the control and high-dose group during the study.

Statistics:

Data of hematology, coagulation, serum biochemistry and urinalysis was collected by Clinical Test Data Administration 1.0. The other data was collected by the Provantis 9.4.3 except nerve function observation and ophthalmological examinations. Where appropriate, quantitative data was analysed by the Provantis 9.4.3 Tables and Statistics Module.
- If > 2 groups, the data were analyzed by Bartlett test for variance homogeneity first. In case of heterogeneity of variance (p<0.05), the Kruskal-Wallis non-parametric analysis of variance for pair wise comparison was used to evaluate the difference between each treated group and the control group. If the test was significant (p<0.05), non-parametric Dunnett's test was then used for pairwise comparison between each treated group and the control group. Statistical analysis was finished when the Kruskal-Wallis test was non-significant (p≥0.05). In the case of homogeneity of variance (p≥0.05), one-way analysis of variance (ANOVA) was used to analyze above parameters. If the test was significant (p<0.05) and the number of animals of each group was the same, Dunnett's test was conducted for pairwise comparison between each treated group and the control group. If the test was significant (p<0.05) and the number of animals of each group was different, Duncan’s test was conducted. Statistical analysis was finished when the test will be non-significant (p≥0.05).
- If 2 groups, pair-wise tests was used by T-test (parametric) or Mann-Whitney U test (non-parametric). The incidences of pathological findings (gross pathological findings, non-graded histopathological findings) was analyzed by the Fisher’s exact test (one-tailed probability level).

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

There were no toxicologically significant effects detected throughout the study. Animals showed hair loss which occurred in both the treatment and control groups and appeared sporadically. So the symptoms noted were considered unrelated to treatment.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

A statistically significant reduction in mean body weight on Day 91 and 98 were noted for the females in 1000 mg/kg dose group only during the recovery period. But no adverse effect was evident in the overall body weight (Day 91-119) and the reduction was considered not to be of toxicological importance. No test item-related body changes were noted. No significant differences were noted in mean body weight or body weight gains during the treatment period for either sex.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

No adverse effect in food consumption was detected in treated animals when compared to controls. Throughout the whole test, including the treatment and recovery phase, animals in the treatment groups showed normal variation in the mean food consumption, incidentally including significant increases or decreases as compared with the control group, but without any toxicological meaning.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Description (incidence and severity):

Ophthalmic examinations were conducted on the high-dose (1000 mg/kg) and control group animals (0 mg/kg) prior to the dosing and at the end of treatment. The results indicated that there were no treatment-related changes in the periocular area of eyes, conjunctiva, iris, cornea, optic disk and blood vessel of the eye fundus.

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

- At the end of the treatment period: Among males, the mean Total Leukocyte counts (WBC) value was statistically significantly increased at 1000 mg/kg (P<0.05, +29.9% vs.controls), but not at 300 mg/kg and only observed in one sex only. Therefore, this was casual without toxicological meaning. Other statistically significant differences were minimal, without any dose relationship.
- At the end of the recovery period: For the males, the high-dose group of 1000 mg/kg showed statistically significant increases in PDW, P-LCR (P <0.05) and RDW-CV, RDW-SD (P<0.01) compared with the untreated control group. For the females, the high-dose group of 1000 mg/kg showed statistically significant increases in RDW-CV and RDW-SD (P<0.05) compared with the control group. These above changes only occurred in the recovery period and were not associated with other indices. No test item related changes were found during the recovery period.
There were no toxicologically significant effects detected in the hematology and coagulation parameters examined.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

At the end of the treatment period: The males at the dose of 300 and 1000 mg/kg showed a significant increase in ALP (+31.3% at P<0.001, +63.6% at P<0.05) compared with the control group. Males in the 100, 300 and 1000 mg/kg dose groups showed a significant decrease in T-Bil, -100%, -95.8% and -129.2% respectively. The females at the dose of 1000 and 300 mg/kg also showed a significant decrease in T-Bil (-90.6% at P<0.001 and -57.6% at P<0.05). The above changes returned to normal after four weeks' withdrawal.
The males in 1000 mg/kg dose group showed a significant decrease in TP and GLB compared with the control group, and in all treated groups showed a significant increase in Urea and A/G, but without a dose-response relationship in the above findings, it was considered there was no toxicological significance.
Females in the 1000 mg/kg dose group showed a significant decrease in GLB (- 8.5% at P<0.05) and a significant increase in A/G (+8.6% at P<0.01). The above changes were minimal and returned to normal after four weeks' withdrawal and it was considered to be a normal biological variation.
At the end of the recovery period: The males at the dose of 1000 mg/kg showed a significant increase in TP and GLB (P<0.05). The males at the dose of 1000 mg/kg showed a significant decrease in A/G (P<0.05). The difference was very small in the males at the end of the recovery period and the results were opposite to the treatment period, it was considered unrelated to the test item. No abnormal findings were observed in the female animals.
(see Table 5 in 'any information on results including tables' for an overview of the results).

Urinalysis findings:

no effects observed

Description (incidence and severity):

There were no statistically significant or treatment-related changes in urinalysis parameters in male or female animals.

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

The results of the behavioural observations during the 11th week of the treatment period indicated that no neurotoxicity was found related to the test item.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Mean absolute and relative liver weights were statistically significantly higher in male rats at 300 and 1000 mg/kg and in female rats at 1000 mg/kg when compared to controls (see Table 4 in 'any information on results including tables'). The liver weights in both male and female animals of the 1000 mg/kg dose groups showed statistically significant in mean absolute increases (+26.3% for the males and +19.2% for the females both at p<0.01) and relative increases (organ-to-body weight ratios: +41.2% for the males at and +26.2% for the females, both at p<0.001; organ-to-brain weight ratios: +30.5% for the males at p<0.001 and +19.2% for the females at p<0.01). The liver weights in male animals of the 300 mg/kg dose groups showed statistically significant in mean absolute increases (+17.1% at p<0.05) and relative increases (organ-to-body weight ratios: +22.7% at p<0.001; organ-to-brain weight ratios: +16.3% at p<0.05). In the male and female test groups of the recovery phase, liver weights were normal without significant deviation from the respective controls.
The liver weights were increased at the highest dose level and the clearly treatment-related effect occurred in both sexes. Although in the absence of any associated histology correlates, the intergroup differences were considered to be of toxicological significance taking into account of the changes of ALP and T-Bil in blood biochemistry.
All other statistically significant organ weight differences were judged to be incidental in view of their individual variation and in the absence of any correlated histopathological finding.

Gross pathological findings:

no effects observed

Description (incidence and severity):

Gross necropsy and macroscopic observation did not provide evidence of any signs of toxicity related to the test item during the whole test.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

There was no evidence of treatment-related microscopic abnormalities.

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Details on results:

Formulation analysis: The concentration of the formulations was determined by analysis on the first dosing day, the 7th week and the 13th week of the dosing, and the actual results for the analysis of the dosing formulations were within ±10% of the nominal concentration.

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

male

Basis for effect level:

clinical biochemistry

organ weights and organ / body weight ratios

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day (nominal)

Based on:

act. ingr.

Sex:

female

Basis for effect level:

clinical biochemistry

organ weights and organ / body weight ratios

Critical effects observed:

not specified

Table 4. Liver weight changes at terminal sacrifice (% change when compared to controls)

Liver weight changes at terminal sacrifice (% change when compared to controls
Sex		Male				Female
Dose level TFSK/TFAK (mg/kg)		0	100	300	1000	0	100	300	1000
Mean absolute liver weight (g) [a]	Mean SD N	13.0354 1.8718 10	13.8423 1.5024 10	15.2701^* 1.4764 10	16.4643^** 2.7111 10	7.6480 0.6255 10	8.5629 1.2161 10	8.3204 1.0144 10	9.1165^** 0.6580 10
Mean liver to body weight ratio (%) [a]	Mean SD N	2.474 0.195 10	2.726 0.210 10	3.034^*** 0.278 10	3.492^*** 0.264 10	2.569 0.159 10	2.892^** 0.155 10	2.951^*** 0.258 10	3.242^*** 0.217 10
Mean liver to brain weight ratio (%) [a]	Mean SD N	587.798 80.178 10	622.568 67.219 10	683.494^* 81.651 10	767.266^*** 111.444 10	372.627 24.254 10	411.884 55.083 10	402.721 48.395 10	444.302^** 38.637 10

  [a] - Anova & Dunnett: * = p < 0.05; ** = p < 0.01; *** = p < 0.001

Table 5. Clinical chemistry parameter changes in males and females (% change when compared to controls)

Sex		Male				Female
Dose level TFSK/TFAK (mg/kg)		0	100	300	1000	0	100	300	1000
ALP [a] (U/L)	Mean SD N %Diff	104.9 18.9 10	126.6 20.4 10 20.7	137.7* 38.0 10 31.3	171.6*** 45.5 10 63.6	52.6 10.7 10	58.3 14.7 10 10.8	54.4 16.0 10 3.4	52.1 7.4 10 -1.0
ALT [a] (U/L)	Mean SD N %Diff	54.4 20.5 10	53.4 12.9 10 -1.8	55.4 14.1 10 1.8	57.7 16 10 6.1	49.1 18.6 10	48.0 13.6 10 -2.2	57.9 33.1 10 17.9	43.1 10.9 10 -12.2
AST [a] (U/L)	Mean SD N %Diff	117.8 55.4 10	124.7 42.6 10 5.9	112.4 19.3 10 -4.6	114.6 27.2 10 -2.7	109.5 29.8 10	106.8 27.2 10 -2.5	125.1 58.7 10 14.2	96.1 32.5 10 -12.2
Urea [a] (mmol/L)	Mean SD N %Diff	5.700 0.823 10	8.371** 4.448 10 46.9	6.950** 0.890 10 21.9	7.323*** 0.769 10 28.5	8.032 2.632 10	8.653 1.215 10 7.7	9.091 2.083 10 13.2	8.913 1.386 10 11.0
Creatinine [a] (µmol/L)	Mean SD N %Diff	29.5 3.6 10	40.3 26.5 10 36.6	32.7 3.0 10 10.8	31.9 4.4 10 8.1	40.1 6.3 10	38.3 5.7 10 -4.5	39.6 4.2 10 -1.2	38.4 6.1 10 -4.2
T-BIL [a] (µmol/L)	Mean SD N %Diff	0.48 0.40 10	0.00** 0.25 10 -100.0	0.02** 0.23 10 -95.8	-0.14*** 0.32 10 -129.2	0.66 0.39 10	0.38 0.21 10 -42.4	0.28* 0.31 10 -57.6	0.06*** 0.23 10 -90.9
GLU [a] (mmol/L)	Mean SD N %Diff	9.348 1.361 10	9.287 2.001 10 -0.7	10.589 1.421 10 13.3	12.281** 2.209 10 31.4	9.132 1.910 10	7.449 1.900 10 -18.4	7.412 1.431 10 -18.8	6.983 2.491 10 -23.5
Triglyceride [a] (mmol/L)	Mean SD N %Diff	0.492 0.190 10	0.510 0.148 10 3.7	0.437 0.095 10 -11.2	0.465 0.058 10 -5.5	0.504 0.191 10	0.456 0.117 10 -9.5	0.473 0.100 10 -6.2	0.477 0.140 10 -5.4
CHO [a] (mmol/L)	Mean SD N %Diff	1.702 0.496 10	1.728 0.243 10 1.5	1.674 0.304 10 -1.6	1.526 0.309 10 -10.3	2.469 0.532 10	2.246 0.296 10 -9.0	2.344 0.314 10 -5.1	2.522 0.349 10 2.1
TP [a] (mmol/L)	Mean SD N %Diff	66.01 2.55 10	64.89 2.21 10 -1.7	65.09 3.24 10 -1.4	61.17** 2.95 10 -7.3	73.61 4.09 10	73.53 4.47 10 -0.1	75.01 3.06 10 1.9	71.02 2.26 10 -3.5
ALB [a] (g/L)	Mean SD N %Diff	41.33 1.51 10	41.88 1.28 10 1.3	41.82 1.49 10 1.2	40.59 0.99 10 -1.8	46.46 2.44 10	46.78 2.67 10 0.7	47.97 2.47 10 3.3	46.17 1.59 10 -0.6
Potassium [a] (mmol/L)	Mean SD N %Diff	6.149 0.459 10	6.049 0.347 10 -1.6	6.184 0.419 10 0.6	6.155 0.396 10 0.1	5.670 0.771 10	6.143 0.832 10 8.3	5.877 0.595 10 3.7	5.554 0.452 10 -2.0
Sodium [a] (mmol/L)	Mean SD N %Diff	149.5 3.3 10	152.6 1.9 10 2.1	151.6 2.1 10 1.4	149.9 3.3 10 0.3	152.0 1.9 10	151.3 2.3 10 -0.5	152.0 1.9 10 0.0	151.9 2.5 10 -0.1
GLB [a] (g/L)	Mean SD N %Diff	24.68 1.33 10	23.01 1.82 10 -6.8	23.27 1.90 10 -5.7	20.58*** 2.21 10 -16.6	27.15 1.89 10	26.75 2.23 10 -1.5	27.04 1.31 10 -0.4	24.85* 1.31 10 -8.5
A/G [a]	Mean SD N %Diff	1.68 0.07 10	1.83** 0.17 10 9.2	1.80** 0.10 10 7.5	1.99*** 0.22 10 18.8	1.71 0.07 10	1.76 0.11 10 2.4	1.78 0.11 10 3.6	1.86** 0.11 10 8.6
Total bile acid [a] (µmol/L)	Mean SD N %Diff	28.18 17.36 10	30.94 35.03 10 9.8	32.61 24.46 10 15.7	37.66 27.57 10 33.6	38.04 17.82 10	38.27 23.57 10 0.6	32.53 17.19 10 -14.5	25.66 7.99 10 -32.5

[a] - Anova & Dunnett: * = p < 0.05; ** = p < 0.01***; = p < 0.001

 

Conclusions:

In the 90-day oral study with Sprague Dawley rats, the reaction mass of TFSK/TFAK was administered daily by oral gavage to male and female rats at 100, 300 and 1000 mg a.i./kg bw/day. The NOAEL for males was considered to be 100 mg/kg, based on marked increase in liver absolute and relative weights and ALP and also decrease in T-Bil at the high and mid dose level. The NOAEL for females was considered to be 300 mg/kg, based on marked increase in liver absolute and relative weights and decrease in T-Bil at the high dose level.

Executive summary:

In a GLP-compliant OECD Guideline 408 study, the reaction mass of TFSK/TFAK was administered daily by oral gavage to separate groups of Sprague Dawley rats (10/sex/group for low and mid dose and 16/sex/group for high dose) at concentrations of 100, 300 and 1000 mg active ingredient/kg bw/day for 90 days. A similarly constituted group of 16 males and 16 females received the vehicle (purified drinking water) and acted as a control. At the end of dosing, surviving animals were sacrificed and subjected to a full gross necropsy, while the residual animals (6 males and 6 females) in the control and high-dose were kept for at least four weeks (the recovery period) without treatment to detect delayed occurrence or persistence of, or recovery from toxic effects.

All animals survived until scheduled sacrifice. No treatment-related changes were noted in clinical appearance, functional observations, body weight, food consumption, haematology, urinalysis and ophthalmoscopy during the study. Dose-related increases were found in ALP for the males at 300 and 1000 mg/kg. Males in all treated groups showed a significant decrease in total bilirubin. Females in the 300 and 1000 mg/kg dose groups also showed a significant decrease in total bilirubin. These changes returned to normal after the four week recovery period.

Dose-related increases in absolute and relative liver weights were found for the 1000 and 300 mg/kg dose groups in males (mean absolute increase of +26.3% and +17.1%, respectively) and for the 1000 mg/kg dose group in females (mean absolute increase of +19.2%). Within the four weeks recovery period, the liver weights regressed to normal.

Based on the results of this study, the no-adverse-observed-effect-level (NOAEL) for the reaction mass of TFSK/TFAK was considered to be 100 mg a.i./kg/day for males, based on marked increase in relative and absolute liver weights and ALP and decrease in T-Bil at the high and mid dose level. The NOAEL for females was considered to be 300 mg a.i./kg/day for females, based on marked increase in relative and absolute liver weights and decrease in T-Bil at the high dose level.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

50 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

GLP-compliant guideline study, klimisch 1

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The repeated dose toxicity studies used for potassium trifluoroacetate evaluation were actually performed with a reaction mass of potassium trifluoroacetate (TFAK) and  potassium trifluoromethanesulphinate (TFSK). Since this reaction mass consists of 50% TFAK, the results are considered appropriate to evaluate the effects after repeated exposure to potassium trifluoroacetate. For the reaction mass of TFSK/TFAK, a combined oral repeated dose toxicity study with reproduction/developmental toxicity screening test performed according to OECD 422 and a sub-chronic oral study according to OECD 408 are available for assessment.

In the 90-day oral study with Sprague Dawley rats, the reaction mass of TFSK/TFAK was administered daily by oral gavage to male and female rats at 100, 300 and 1000 mg a.i./kg bw/day. The NOAEL for males was considered to be 100 mg/kg, based on marked increase in liver absolute and relative weights and ALP and also decrease in T-Bil at the high and mid dose level. The NOAEL for females was considered to be 300 mg/kg, based on marked increase in liver absolute and relative weights and decrease in T-Bil at the high dose level. Therefore, the actual NOAEL for TFAK is considered to be 50 mg/kg bw/day.

In addition, for the reaction mass of TFSK/TFAK, a combined oral repeated dose toxicity study with reproduction/developmental toxicity screening test performed according to OECD 422. In this GLP compliant study, the test item was administered daily by gavage to male and female Sprague Dawley rats, at dose levels of 100, 300 and 1000 mg/kg/body weight/day, for approximately 38 days for males and at maximum 57 days for females. In parent animals, the test item administration at 300 and 1000 mg/kg/day active ingredient induced centrilobular hepatocellular hypertrophy. This microscopic change correlated with increased liver weights. This change was not considered to be adverse in view of the absence of associated degenerative microscopic findings or clinical pathology changes in liver enzyme activities. Based on these results, the general NOAEL (No Adverse Observed Effect Level) was considered to be 1000 mg/kg/day. In this study, about 50 % (49.9 %) of the active ingredients is TFAK, the remaining being TFSK. Therefore, the actual NOAEL for TFAK is considered >500 mg/kg bw/day.

 

Justification for classification or non-classification

Based on the available repeated dose toxicity studies, the test substance does not meet the criteria of the EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008 and therefore no classification is needed.