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Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
End on 11 JULY 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: study performed according to OECD guideline and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Vegeflux soy
- Substance type: UVCB
- Physical state: viscous liquid
- Stability under test conditions: data not available
- Storage condition of test material: at room temperature

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Animal House, SGS Life Science Services
- Age at study initiation: 6 - 8 weeks
- Weight at study initiation: 216 to 272 g (day 0)
- Fasting period before study: data not available
- Housing: animals were housed group wise (5 animals/sex/cage) in standard polypropylene cages with stainless steel top grill
- Diet: rat pellet feed (Amrut brand), ad libitum
- Water: potable water, ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19.9 - 23.1 °C
- Humidity: 59 - 69%
- Air changes (per hr): data not available
- Photoperiod (hrs dark / hrs light): data not available

IN-LIFE DATES: from 03 APRIL 2008 to 30 MAY 2008

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5% methyl cellulose
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: the test item was prepared afresh daily with 0.5% methyl cellulose

VEHICLE
- Justification for use and choice of vehicle (if other than water): data not available
- Concentration in vehicle: data not available
- Amount of vehicle (if gavage): 10 mL/kg body weight
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
28 days
Frequency of treatment:
daily, 7 days each week
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 150, 450, 1000 mg/kg bw
Basis:
other: nominal
No. of animals per sex per dose:
5 animals/sex/group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on a pilot study at 1000 and 1500 mg/kg bw for 7 days
- Rationale for animal assignment: animals were assigned to the different groups using a randomization procedure
- Post-exposure recovery period in satellite groups: 14 days in 2 satellite groups (0 and 1000 mg/kg, 10 animals/group)
Positive control:
not applicable

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: animals were observed daily

BODY WEIGHT: Yes
- Time schedule for examinations: body weight was recorded prior to treatment and at weekly interval thereafter

FOOD CONSUMPTION: calculated at weekly interval throughout the study

FOOD EFFICIENCY: not determined

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: prior to the first treatment and prior to sacrifice (using ophtalmoscope)
- Dose groups that were examined: all animals

HAEMATOLOGY: Yes
- Time schedule for collection of blood: blood samples were collected at the end of dosing or post-treatment observation period
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: all animals
- Parameters checked in table 1 were examined: haemoglobin, RBC, WBC (total and differential), haematocrit (HCT), platelet count, MCH, MCHC, MCV and clotting time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of dosing and post-treatment observation period
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked in table 1 were examined: glucose (fasted), urea, creatinine, total protein, albumin, total bilirubin, Alanine aminotransferase, Aspartate aminotransferase, sodium, cholesterol and potassium

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: during the 4th week of dosing or during the 2nd week of post-treatment observation period
- Dose groups that were examined:all animals
- Battery of functions tested: sensory reactivity / grip strength / motor activity
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table 2)

HISTOPATHOLOGY: Yes (see table 2)
Statistics:
the change in body weight, feed consumption, haematology, clinical chemistry, and organ weights of animals belonging to the treatment groups were compared to the control group by statistical analysis using Student's 't'-test.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
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 examined
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
- CLINICAL SIGNS AND MORTALITY: No mortality occurred in any of the treatment and control group. However loose stools were observed after test item administration in all groups of treated and control animals that persisted up to 5th day of observation period. The sign exhibited by the animals could not be related to test item administration and it may be due to the vehicle used.

BODY WEIGHT AND WEIGHT GAIN: statistically significant changes were observed on second and thirD week of 1000 mg/kg bw/day (G5) and third week of 450 mg/kg bw/day (G4) in the male body weight when compared with control groups of animals.

FOOD CONSUMPTION: No significant changes were observed in feed consumption data of control and treated groups.

HAEMATOLOGY: Significant changes were observed on 29th day in haematological parameters such as clotting time, MCV (G6 high dose reverse males - 1000 mg/kg bw/day) and MCHC (G3 low dose males - 150 mg/kg bw/day) when compared with the control (G1 and G2) groups. However these variations were observed to be within the biological range of these species. No significant changes were observed on the 43th day.

CLINICAL CHEMISTRY: Statistically significant changes were observed on 29th day in biological parameters such as AST (G5-1000 mg/kg bw/ day and G6 high dose reversal - 1000 mg/kg bw/ day), total protein (G6) and cholesterol (G6). However these variations were observed to be within the biological range of this species. No significant changes were observed on the 43th day.

ORGAN WEIGHTS: No significant changes were observed in absolute and relative organ weights of control and treated groups.

GROSS PATHOLOGY AND HISTOPATHOLOGY: No test item related toxicologically significant and histopathological lesions were recorded in any of the treated groups of animals when compared to control groups.

Effect levels

Dose descriptor:
NOEL
Effect level:
> 1 000 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: (no effect observed)

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 3: summary of weekly mean body weights (g)

Group

Sex: male

Week

 

0

1

2

3

4

5

6

G1 – Control

223

240

271

291

312

 

 

G2 –Control Reversal

221

243

265

286

303

322

336

G3 – 150 mg/kg bw

220

244

267

287

307

 

 

G4 – 450 mg/kg bw

219

240

260

280 *

299

 

 

G5 – 1000 mg/kg bw

217

237

258 *

278 *

303

 

 

G6 – 1000 mg/kg bw – Reversal

216

236

262

282

302

323

338

 

Sex: Female

G1 – Control

272

284

293

303

312

 

 

G2 –Control Reversal

266

279

292

304

316

325

332

G3 – 150 mg/kg bw

256

269

280

291

299

 

 

G4 – 450 mg/kg bw

256

267

277

290

300

 

 

G5 – 1000 mg/kg bw

264

275

288

299

309

 

 

G6 – 1000 mg/kg bw – Reversal

267

279

291

303

314

326

336

*: significant when compared with control group (P<0.05)

 

Table 4: summary of clinical chemistry parameters Day-29

Group

Total protein (g/dl)

Albumin (g/dl)

Glucose (mg/dl)

AST (IU/l)

ALT (IU/l

Urea (mg/dl)

Creatinine (mg/dl)

Cholesterol (mg/dl)

Total bilirubine (mg/dl)

Sodium (mEq/L)

Potassium (mEq/L)

Sex: Male

G1 control

6.62

1.22

115.60

103.4

49.00

4.33

0.38

85.40

0.14

143.00

3.53

G2 control reversal

6.76

1.24

105.20

136.0

50.80

6.45

0.58

74.80

0.16

143.00

3.70

G3 150 mg/kg

7.12

1.30

121.00

111.8

53.20

4.17

0.42

99.20

0.18

145.40

3.78

G4 450 mg/kg

7.36

1.30

119.40

112.6

55.20

7.00

0.52

99.40

0.16

143.20

3.71

G5 1000 mg/kg

6.94

0.98

119.60

121.6*

54.20

5.62

0.40

95.00

0.14

145.80

3.66

G6 1000 mg/kg reversal

6.62

1.28

116.00

120.0

53.00

3.82

0.48

99.40 *

0.16

144.80

3.93

 

Sex: Female

G1 control

6.86

1.10

115.80

119.4

47.80

8.10

0.48

98.00

0.18

142.80

3.71

G2 control reversal

7.04

1.42

119.00

117.6

52.60

5.50

0.50

94.80

0.12

142.40

3.70

G3 150 mg/kg

7.04

1.24

126.40

113.4

52.40

7.37

0.46

95.40

0.14

141.80

3.61

G4 450 mg/kg

6.84

1.28

111.40

130.2

45.60

8.27

0.50

100.80

0.12

144.20

3.51

G5 1000 mg/kg

6.66

1.30

114.40

127.6

48.60

7.78

0.42

106.80

0.14

143.20

3.54

G6 1000 mg/kg reversal

6.68 *

1.36

116.80

110.4 *

50.20

6.27

0.52

103.80

0.14

145.40

3.50

*: significant when compared with control group (P<0.05)

Table5: summary of haematological parameters Day-29

Group

RBC (106/µl)

Hb (g/dl)

HCT (%)

WBC (106/µl)

PLT (103/µl)

CT (Sec)

L (%)

M (%)

G (%)

MCH (pg)

MCHC (g/dl)

MCV (fl)

 

Sex: Male

G1 control

9.37

11.54

53.86

12.02

854.80

94.20

77.09

6.00

16.91

12.40

21.54

58.26

G2 control reversal

8.34

13.30

57.44

10.75

895.20

133.20

75.44

6.30

18.26

16.11

23.34

69.72

G3 150 mg/kg

9.49

12.42

48.96

8.65

1033.80

103.60

74.12

7.00

18.88

13.16

25.46 *

51.79

G4 450 mg/kg

10.02

12.72

59.24

10.53

949.80

102.00

81.06

2.98

15.96

12.76

21.50

59.35

G5 1000 mg/kg

9.37

11.24

59.24

8.65

1033.80

96.20

78.58

4.72

16.70

12.04

19.06

63.88

G6 1000 mg/kg reversal

9.75

13.26

52.78

10.06

949.80

101.00 *

74.94

8.54

16.52

13.75

25.20

54.52

 

Sex: Female

G1 control

8.52

11.62

51.56

11.71

882.60

84.60

69.62

4.62

25.76

13.79

22.68

61.46

G2 control reversal

9.06

13.08

52.86

14.88

746.40

100.60

73.08

6.36

20.56

14.48

25.02

58.39

G3 150 mg/kg

8.81

11.40

48.96

10.68

922.40

90.80

74.27

4.68

21.05

13.15

23.38

56.56

G4 450 mg/kg

8.73

11.44

52.60

10.01

952.40

93.20

74.69

6.66

18.65

13.11

21.91

60.46

G5 1000 mg/kg

8.49

11.40

52.60

10.25

953.60

88.80

78.58

3.24

18.18

13.60

21.88

63.26

G6 1000 mg/kg reversal

8.67

12.26

50.91

11.92

1088.60

103.20

70.80

8.54

20.66

14.24

24.29

59.04

*: significant when compared with control group (P<0.05)

Table 6: summary of relative organ weights (g)

Group

Adrenal

Thymus

Spleen

Heart

Brain

Kidney

Liver

Sex: Female

G1 control

0.011

0.130

0.290

0.270

0.573

0.516

2.763

G2 control reversal

0.013

0.127

0.273

0.260

0.552

0.542

2.801

G3 150 mg/kg

0.012

0.124

0.311

0.285

0.611

0.577

2.766

G4 450 mg/kg

0.013

0.136

0.316

0.290

0.603

0.540

2.954

G5 1000 mg/kg

0.016

0.117

0.307

0.271

0.593

0.530

2.536

G6 1000 mg/kg reversal

0.009

0.106

0.278

0.257

0.544

0.517

2.525

 

Group

Adrenal

Epididymes

Thymus

Spleen

Heart

Brain

Testes

Kidney

Liver

Sex: Male

G1 control

0.012

0.306

0.116

0.366

0.313

0.596

0.965

0.603

3.118

G2 control reversal

0.011

0.277

0.110

0.348

0.285

0.572

0.861

0.564

3.006

G3 150 mg/kg

0.011

0.296

0.131

0.440

0.319

0.630

0.978

0.601

3.153

G4 450 mg/kg

0.015

0.315

0.126

0.424

0.313

0.628

0.974

0.623

3.244

G5 1000 mg/kg

0.011

0.302

0.140

0.425

0.306

0.619

0.939

0.617

3.119

G6 1000 mg/kg reversal

0.011

0.269

0.115

0.365

0.284

0.575

0.863

0.556

2.865

Applicant's summary and conclusion

Conclusions:
Based on the above observations, the NOEL of Vegeflux Soy for Wistar rats can be considered as greater than 1000 mg/kg bw/day.
Executive summary:

In a subacute toxicity study (Thanikaivel V, 2008), Vegeflux Soy was administered to 5 Wistar rats/sex/dose by gavage at dose levels of 0, 150, 450 and 1000 mg/kg bw/day for 28 days. The test was performed according to OECD guideline 407 and GLP. This subacute toxicity study in the rat is considered as acceptable.

 

There were no compound related effects in mortality, clinical signs, body weight, food consumption, haematology, clinical chemistry, organ weights, or gross and histological pathology.

Based on these results, the NOEL of Vegeflux Soy is greater than 1000 mg/kg bw/day.