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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
The aim of this study was to assess the toxicity potential of test chemical after single oral administration in rats.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report):4,4'-methylenedi-2,6-xylenol
- Molecular formula :C17H20O2
- Molecular weight :256.34 g/mol
- Substance type:Organic
- Physical state:White Solid (Powder)
- Analytical purity:99.79%
- Lot/batch No.:Lot 1/03
- Storage condition of test material:Stored in cool, dry place. Kept container closed when not in use.
- Other:
Handling and Disposal
Safety precautions: Avoided excessive heat and light. Did not breath dust. Aprons, caps, gloves and goggles were used to ensure the health and safety of the Personnel.
Disposal:The remaining unused test item formulations were disposed as per internal SOPs of sa-FORD.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Bharat Serum and Vaccines Limited.
- Age at study initiation:8- 11 weeks at the time of dosing.
- Health Status :Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
- Weight at study initiation:Minimum: 183 g and Maximum: 216 g (Individual body weights were within ± 5% prior to treatment after overnight fasting).
- Fasting period:The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing.
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding:All cages were provided with corn cobs.
- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle:All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum):All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum.
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period:Animal nos. 1-3 were acclimatized for five days, 4-6 for seven days, 7-9 for nine days and 10-12 for six days prior to administration of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 20.00 °C and Maximum: 23.60 °C.
- Humidity (%):Minimum: 37.40% and Maximum: 61.80%
- Air changes (per hr):More than 12 changes per hour.
- Photoperiod (hrs dark / hrs light):12:12

Administration / exposure

Route of administration:
oral: unspecified
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Amount of vehicle:10 ml
- Justification for choice of vehicle:Corn oil was selected because test item was not soluble in distilled water.
- Lot/batch no. (if required):MKBD4650
- Purity:N/A

MAXIMUM DOSE VOLUME APPLIED:10 ml/kg body weight.

Doses:
G1 = 300 mg/kg bw
G2 = 2000 mg/kg bw
No. of animals per sex per dose:
Twelve female rats
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Daily
- Necropsy of survivors performed: yes
At the end of 14 day observation period, all the survived rats were euthanised by overdose of CO2 for external and internal observations.
- Other examinations performed:
Clinical Observation
After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Additionally, on day 0 animal nos. 7-9 were observed at 6 hours post dosing. Subsequently, all the animals were observed once a day during the 14 day observation period.

Body weight
All rats were weighed on days 0 (prior to dosing), 7 and 14. Animals were weighed immediately after found dead.

other:
Mortality
All the animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: Non toxic to rat.
Mortality:
No mortality was observed in the animals treated with 300 and 2000 mg/kg dose throught out the 14 days observation period post dosing.
Clinical signs:
At 300 mg/kg, all the six animals (animal nos. 1-6) were observed normal throughout experimental period. At 2000mg/kg, animal no. 7 was observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3, 4 and 6 hours and mild ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 8 was observed normal at 30 minutes and 1 hour post dosing, mild to severe lethargy at 2, 3, 4 and 6 hours and mild to moderate ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 9 was observed normal at 30 minutes and 1 hour post dosing, moderate to severe lethargy and mild to moderate ataxia at 2, 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal nos. 10-12 were observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3 and 4 hours and mild ataxia at 3 and 4 hours followed by normal clinical sign till day 14.
Body weight:
Mean body weight was observed with gain was observed in the animals treated with 300 and 2000 mg/kg body weight, during the 14 day observation period, as compared to day 0.
Gross pathology:
No external and internal gross pathological changes were seen in all the six animals treated with 300 and 2000 mg/kg body weight during terminal sacrifice

Any other information on results incl. tables

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day

0-7

Day

0-14

1

G1/ 300

210

223

234

6.19

11.43

2

204

229

234

12.25

14.71

3

200

230

237

15.00

18.50

4

216

240

253

11.11

17.13

5

204

240

246

17.65

20.59

6

198

237

218

19.70

10.10

7

G2/ 2000

210

219

229

4.29

9.05

8

197

228

224

15.74

13.71

9

195

220

235

12.82

20.51

10

187

217

229

16.04

22.46

11

183

219

228

19.67

24.59

12

185

215

229

16.22

23.78

 

Table 2: Summary of Animal Body Weight (g) and Body Weight Changes (%)

 

Sex:Female

Group/ Dose (mg/kg)

Rats Body Weight (g)

Body Weight Changes (%)

Day 0

Day 7

Day 14

0-7

0-14

G1/ 300

Mean

205.33

233.17

237.00

13.65

15.41

SD

6.65

6.91

11.97

4.87

4.10

n

6

6

6

6

6

G2/ 2000

Mean

192.83

219.67

229.00

14.13

19.02

SD

10.09

4.46

3.52

5.29

6.25

n

6

6

6

6

6

Keys:SD = Standard Deviation, n = Number of Animals


Table 3: Individual Animal Clinical Signs and Symptoms

 

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Hours (Day 0)

1/2

1

2

3

4

6

1

G1/ 300

1

1

1

1

1

-

2

1

1

1

1

1

-

3

1

1

1

1

1

-

4

1

1

1

1

1

-

5

1

1

1

1

1

-

6

1

1

1

1

1

-

7

G2/ 2000

1

1

99+

99++

21+

99++

21+

99++

21+

8

1

1

99+

99++

21+

99+++

21+

99+++

21++

9

1

1

99++

21+

99+++

21++

99+++

21++

99+++

21++

10

1

1

99+

99++

21+

99++

21+

-

11

1

1

99+

99++

21+

99++

21+

-

12

1

1

99+

99++

21+

99++

21++

-

Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
Under the conditions of this; acute oral toxicity study of test chemical in female rats is as given below:
The acute oral LD50 value of test chemical was 5000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute oral toxicity i.e it is acutely non toxic to animals.
Executive summary:

This study was performed as per OECD No. 423. Twelve female Wistar rats were selected for acute oral toxicity study.

The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing, with food withheld but drinking water providedad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs.

Three rats of group G1 were dosed with starting dose of 300 mg/kg body weight and the animals did not show any mortality so another three animals of the group G1 were dosed with 300 mg/kg body weight and no mortality was observed. Three animals of the group G2 were dosed with 2000 mg/kg body weight and no mortality was observed, therefore another three animals of same group G2 were dose with 2000 mg/kg and no mortality was observed. Hence, further dosing was stopped.

Body weights were re­corded on day 0 (prior to dosing) 7 and 14. Mean body weight was observed with gain was observed in the animals treated with 300 and 2000 mg/kg body weight, during the 14 day observation period, as compared to day 0.

At 300 mg/kg, all the six animals (animal nos. 1-6) were observed normal throughout experimental period. At 2000mg/kg, animal no. 7 was observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3, 4 and 6 hours and mild ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 8 was observed normal at 30 minutes and 1 hour post dosing, mild to severe lethargy at 2, 3, 4 and 6 hours and mild to moderate ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 9 was observed normal at 30 minutes and 1 hour post dosing, moderate to severe lethargy and mild to moderate ataxia at 2, 3, 4 and 6 hours followed by normal clinical sign till day 14.Animal nos. 10-12 were observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3 and 4 hours and mild ataxia at 3 and 4 hours followed by normal clinical sign till day 14.

No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice.

The acute oralL D50 value of test chemical was 5000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute oral toxicity i.e it is acutely non toxic to animals.