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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
repeated dose toxicity: oral
Remarks:
other: 7 day range-finding study
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
9 October 2012 to 01 November 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: A non-GLP study performed to sound scientific principles with a sufficient level of detail to assess the quality of the relevant results.

Data source

Reference
Reference Type:
other: Study notes from range-finding test
Title:
Unnamed
Year:
2012
Report Date:
2012

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Groups of 3 rats per sex per dose were exposed to the test material in a 7 day palatability test, designed to yield information to determine the dose range for an upcoming 14 day dietary dose range finding study. Animals received treated diet over a 7 day period at the following nominal dose levels; 0 (control), 140, 400, 1200 and 4000 ppm. Mortality, clinical signs, body weight, food consumption and water consumption were monitored during the study. Following the last day of treatment the animals were sacrificed and subjected to gross pathology.
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material (as cited in study report): dipotassium heptafluorotantalate
- Appearance: white crystalline powder
- Storage condition of test material: controlled room temperature (15 - 25 °C, below 70 RH%)

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Crl:WI
- Age at study initiation: at least 8 weeks.
- Weight at study initiation: 233 - 281 g (males); 191 - 215 g (females). Weights did not exceed ± 20 % of the mean weight for each sex at onset of treatment.
- Housing: animals were housed in groups of up to 3 animals per sex per goroup, in grid floor cages.
- Diet: Complete breeding and maintenance diet for rats and mice was provided, ad libitum.
- Water: tap water, ad libitum.
- Acclimation period: at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 15 - 20 air changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light.

Administration / exposure

Route of administration:
oral: feed
Vehicle:
other: sunflower oil
Details on oral exposure:
DIET PREPARATION
4 g of test material was mixed with 50 g (approximately 60 mL) of sunflower oil. This formulation was then mixed into a diet premix with powdered diet before being mixed into a total weight of 1 kg of powder diet mix. This procedure was used to prepare the 4000 ppm diet, the maximum concentration used in the study.
Animals dosed at 0 ppm (control group) received powder diet mixed with 5 % w/w sunflower oil.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
At least three samples of test material treated diets were collected into plastic tubes (from the top, middle and bottom of the container) on day 0, (for homogeneity) and day 7. At least one sample will be similarly taken from the control diet on each occasion.
The level of test material in the diet was determined by ICP.
Duration of treatment / exposure:
7 days
Frequency of treatment:
Continuously (in diet)
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
0, 140, 400, 1200, 4000 ppm
Basis:
nominal in diet
Remarks:
Doses / Concentrations:
0, 13, 39, 117, 350 mg/kg bw/day
Basis:
other: target dose levels, achieved dose levels dependent on food consumption.
No. of animals per sex per dose:
3 males and 3 females per dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: doses were selected, based on the results of previous investigations on the test material in attempt to obtain palatability information and identify toxicity associated with the test material under the conditions of this study and to permit the selection of the intended high dose level for use in a long term repeated dose toxicity study.
- Rationale for animal assignment: Animals were sorted according to body weight by computer and divided to weight ranges. There were an equal number of animals from each weight group randomly assigned to each dose group to ensure that animals of all test groups were of a uniform weight. The grouping was controlled by SPSS/PC software, according to the actual body weight verifying the homogeneity/variability between/within the groups and cages. Males and females will be randomised separately.

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Animals were inspected for signs of morbidity and mortality twice daily (at the beginning and end of each working day.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: At least twice daily, animals may be observed more frequently in cases that show signs of toxicity. The onset, degree and duration of all signs were recorded as applicable.
- Signs evaluated included: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e.g. lachrymation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), and bizarre behaviour (e.g. self-mutilation, walking backwards). Special attention was directed towards the observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.

BODY WEIGHT: Yes
- Time schedule for examinations: The body weight of each animal was recorded on Day 0, 2, 4 and 6 (prior to necropsy).

FOOD CONSUMPTION: Yes
- Time schedule: Daily

WATER CONSUMPTION: Yes
- Time schedule for examinations: Daily, estimated by weighing the daily consumption by cage.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Gross necropsy was performed on each animal irrespective of the date of death, including the animals found dead or euthanised pre-terminally in extremis. Surviving animals were euthanised under pentobarbital anaesthesia by exsanguination.
After exsanguination the external appearance was examined, cranium, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed macroscopically. Any abnormality was recorded with details of the location, colour, shape and size, as appropriate. Special attention was paid to the gastrointestinal tract (stomach mucosa and full length of intestine).

On completion of the macroscopic examination, stomach, intestine, kidneys and any tissues showing macroscopic abnormality were preserved in 10 % buffered formaldehyde solution or in Bouin’s fixative solution (testes and epididymides) or Davidson’s fixative solution (eyes).
Statistics:
Statistical evaluation of data was performed with the program package SPSS PC+4.0 (SPSS Hungary Kft, Budapest). The homogeneity of variance between groups was checked by Bartlett`s homogeneity of variance test. Where no significant heterogeneity was detected a one-way analysis of variance (ANOVA) was made. If the obtained result was significant Duncan’s Multiple Range test was used to assess the significance of inter-group differences. Significant results with inter-group comparisons were further compared using Kruskal-Wallis and Mann-Whitney U-tests.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
See "Details on results" for information.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
See "Details on results" for information.
Histopathological findings: non-neoplastic:
not examined
Histopathological findings: neoplastic:
not examined
Details on results:
MORTALITY
None of the animals died during the study.

BODY WEIGHT AND WEIGHT GAIN
Decrease in body weight gain was observed in males and females, but this was not unequivocally dose-related.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
There was no negative effect recorded in food consumption, however, spillage of the diet was observed in all groups (including the control group). Calculation of food consumption (and test material intake) was therefore not possible.

GROSS PATHOLOGY
Gross pathology revealed significant effects on the stomach mucosa, as follows:
- Concentration 4000 ppm: ulcers, glandular mucosa, dark red focuses on mucosa
- Concentration 1200 ppm: ulcers, glandular mucosa, dark red focuses on mucosa
- Concentration 400 ppm: ulcers, glandular mucosa
- Concentration 140 ppm: no effect, NOAEL under the conditions of this study.

Effect levels

Dose descriptor:
NOAEL
Effect level:
140 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Based on significant effects observed in the stomach and mucosa.

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1: Body Weight and Weight Gain

Concentration (mg/kg diet)

Sex

Individual body weights

Individual body weight gain from Day 0 to Day 7

Mean body weight gain Day 0 to Day 7

Day 0

Day 2

Day 4

Day 6

 Day 7

0

Male

251

264

265

282

286

35

50.33

246

263

273

294

304

58

250

276

281

303

308

58

Female

200

198

206

216

223

23

27.00

194

211

210

226

223

29

201

215

217

231

230

29

140

Male

257

268

274

291

291

34

49.00

259

278

288

308

318

59

233

248

256

278

287

54

Female

205

218

218

224

219

14

17.67

203

209

217

219

222

19

203

203

213

214

223

20

400

Male

254

268

281

301

315

61

57.67

239

257

261

283

291

52

264

278

290

311

324

60

Female

202

218

224

235

236

34

23.67

192

200

206

210

214

22

200

197

211

208

215

15

1200

Male

253

262

271

290

298

45

54.00

245

260

269

292

306

61

273

287

297

317

329

56

Female

193

208

209

218

211

18

22.00

208

222

225

236

230

22

191

206

202

212

217

26

4000

Male

281

268

281

185

298

17

0.67

248

255

261

280

197

-51

251

246

257

268

287

36

Female

200

199

197

211

216

16

12.00

215

212

211

223

230

15

195

188

194

186

200

5

 

Table 2: Food Consumption

Concentration (mg/kg diet)

Sex

Food consumption (g/animal/day)*

Mean food consumption g/ animal/Day

Day 0

Day 1

Day 2

Day 3

Day 4

Day 5

Day 6

0

Male

28

29

27

30

27

26

28

27.86

Female

23

22

12

24

23

21

23

21.14

140

Male

27

28

27

32

27

24

28

27.57

Female

21

21

26

26

22

17

28

23.00

400

Male

30

29

26

31

30

27

31

29.14

Female

26

26

20

29

21

21

25

24.00

1200

Male

27

28

28

31

38

28

31

30.14

Female

26

33

22

35

24

28

21

27.00

4000

Male

19

22

25

39

25

32

41

29.00

Female

23

27

27

39

23

25

42

29.43

* calculated data = cage level measured food consumption/number of animals in the cage (3)

Applicant's summary and conclusion

Conclusions:
Under the conditions of the study none of the animals died. Decrease in body weight gain was observed in males and females, but this was not unequivocally dose-related. There was no negative effect recorded in food consumption, however, spillage of the diet was observed in all groups (including the control group). Calculation of food consumption (and test material intake) was therefore impaired. Gross pathology revealed significant effects on the stomach mucosa, most probably due to degradation of the test material in the stomach to Ta2O5, KF and HF. KF and HF are considered responsible for acute corrosive effects seen in the gastrointestinal tract.
Executive summary:

The repeated dose toxicity, and palatability, of the test material was investigated in a preliminary study in which groups of 3 rats per sex per dose were exposed to the test material over 7 consecutive days. Animals received treated diet at the following nominal dose levels; 0 (control), 140, 400, 1200 and 4000 ppm formulated in sunflower oil. Mortality, clinical signs, body weight, food consumption and water consumption were monitored during the study. Following the last day of treatment the animals were sacrificed and subjected to gross pathology.

None of the animals died. Decrease in body weight gain was observed in males and females, but this was not unequivocally dose-related. There was no negative effect recorded in food consumption, however, spillage of the diet was observed in all groups (including the control group). Calculation of food consumption (and test material intake) was therefore impaired. Gross pathology revealed significant effects on the stomach mucosa, most probably due to degradation of the test material in the stomach to Ta2O5, KF and HF. KF and HF are considered responsible for acute corrosive effects seen in the gastrointestinal tract.

Under the conditions of the study the No Observed Adverse Effect Level (NOAEL) was considered to be 140 ppm in diet.