Registration Dossier

Toxicological information

Acute Toxicity: oral

Currently viewing:

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:
comparable to guideline study
Justification for type of information:
data is from experimental reports

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 acute oral toxicity study of the given test chemical was performed in rat.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
SOURCE OF TEST MATERIAL
- Identity: Reaction mass of 4-(2,6,6-trimethylcyclohex-2-ene-1-yl)-but-3-ene-2-one and -(2,6,6-trimethylcyclohex-1-ene-1-yl)-but-3-ene-2-one 100%
- EC no: 907-706-6
- Lot/batch No.of test material: 9000533813
- Expiration date of the lot/batch: April 14, 2004
- Purity: 85.1% (sum of 2 isomers)
- Colour: colourless to pale yellow

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing:On the day of the experiment, the test item was dissolved in ethanol (purity > 99 %,MERCK, D-64293 Darmstadt).

Specific details on test material used for the study:
- Name of test material: Reaction mass of 4-(2,6,6-trimethylcyclohex-2-ene-1-yl)-but-3-ene-2-one and -(2,6,6-trimethylcyclohex-1-ene-1-yl)-but-3-ene-2-one
- Molecular formula: C13H20O
- Molecular weight: 192.3 g/mole
- Smiles : C1([C@@H](C(=CCC1)C)\C=C\C(C)=O)(C)C
- Inchl: 1S/C13H20O/c1-10-6-5-9-13(3,4)12(10)8-7-11(2)14/h6-8,12H,5,9H2,1-4H3/b8-7+
- Substance type: Organic
- Physical state: Liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: In-house animals, bred at Animal House, sa-FORD.CPCSEA Registration No. 1256/bc/09/CPCSEA.
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: Female rats of the age of approximately 8 to 9 weeks old were used.
- Weight at study initiation: Minimum: 129 g Maximum: 143 g (Individual body weights were within ± 4% prior to treatment after overnight fasting)
- Fasting period before study: The food was withheld prior to dosing and 3-4 hours post dosing but drinking water was provided ad libitum.
- Housing: The animals were housed individually in polycarbonate cages.All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) SPAR – 24/ 2013.
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No.: 400012.
- 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, prior to administration of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Room temperature was maintained at 21.00 to 24.40 degree centigrade.
- Humidity (%): Room humidity was maintained at 36.60% to 57.30%.
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2000mg/kg bw
- Amount of vehicle (if gavage):10ml
- Justification for choice of vehicle: Test material soluble in corn oil
- Lot/batch no. (if required): MKBD4650

MAXIMUM DOSE VOLUME APPLIED: The maximum dose volume administered was 10 ml/kg body weight.
DOSAGE PREPARATION (if unusual): The dosing solution was prepared fresh, prior to dose administration.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Dose 2000 mg/kg:6 female
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: once daily
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical Observations and General Appearance: 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). Subsequently, all animals were observed once a day during the 14 day observation period.
Mortality: All animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.
Body weights: All rats were weighed on days 0 (prior to dosing), 7 and 14.
Gross Pathology: At the end of 14 day observation period, all the survived rats were euthanised by overdose of CO2 for external and internal observations.
Statistics:
not specified

Results and discussion

Preliminary study:
not specified
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
No mortality was observed in the animals treated with 2000 mg/kg dose throught out the 14 days observation period
Clinical signs:
At 2000 mg/kg, animal nos. 1, 4, 5 and 6 were observed normal at 30 minutes, 1 hour, on day 1 to 14 post dosing and moderate to mild lethargy was observed at 2, 3 and 4 hours post dosing. Animal nos. 2 and 3 were observed normal at 30 minutes, 1 hour, on day 1 to 14 post dosing and mild lethargy was observed at 2, 3 and 4 hours post dosing
Body weight:
Body weight gain was observed in all surviving animals treated with 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 the animals treated with 2000 mg/kg body weight during terminal sacrifice
Other findings:
not specified

Any other information on results incl. tables

Individual Animal Body Weight (g) andBody Weight Changes(%)

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day

0-7

Day

0-14

1

G1/ 2000

143

176

187

23.08

30.77

2

138

170

187

23.19

35.51

3

141

169

185

19.86

31.21

4

129

155

167

20.16

29.46

5

142

155

179

9.15

26.06

6

134

169

174

26.12

29.85

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/ 2000

Mean

137.83

165.67

179.83

20.26

30.47

SD

5.42

8.66

8.11

5.90

3.06

n

6

6

6

6

6

Keys:- = Not Applicable, SD = Standard Deviation, n = Number of Animals

Individual Animal Clinical Signs and Symptoms

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Hours (Day 0)

1/2

1

2

3

4

1

G1/ 2000

1

1

99++

99++

99+

2

1

1

99+

99+

99+

3

1

1

99+

99+

99+

4

1

1

99++

99+

99+

5

1

1

99++

99+

99+

6

1

1

99++

99+

99+

Animal No.

Group/ Dose (mg/kg)

Days post dosing

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1

G1/ 2000

1

1

1

1

1

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

1

1

1

1

1

1

4

1

1

1

1

1

1

1

1

1

1

1

1

1

1

5

1

1

1

1

1

1

1

1

1

1

1

1

1

1

6

1

1

1

1

1

1

1

1

1

1

1

1

1

1

Keys:1 = Normal, 99 = Lethargy, + = Mild, ++ = Moderate

Individual Animal Mortality Record

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Day of Observation (Day 0 to 14)

Morning Observations

Evening Observations

1

G1/ 2000

No mortality and morbidity

No mortality and morbidity

2

No mortality and morbidity

No mortality and morbidity

3

No mortality and morbidity

No mortality and morbidity

4

No mortality and morbidity

No mortality and morbidity

5

No mortality and morbidity

No mortality and morbidity

6

No mortality and morbidity

No mortality and morbidity

Gross Necropsy Observation

 Sex:Female

Animal No.

Group/ Dose (mg/kg)

Mode of Death

Gross Observation

External

Internal

1

G1/ 2000

Terminal sacrifice

No abnormality detected

No abnormality detected

2

Terminal sacrifice

No abnormality detected

No abnormality detected

3

Terminal sacrifice

No abnormality detected

No abnormality detected

4

Terminal sacrifice

No abnormality detected

No abnormality detected

5

Terminal sacrifice

No abnormality detected

No abnormality detected

6

Terminal sacrifice

No abnormality detected

No abnormality detected


Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
The LD50 value was considered to be >2000 mg/kg body weight, when female wistar rats were treated with the given test chemical orally.
Executive summary:

The acute oral toxicity study of the given test chemical was performed as per OECD 423 guideline in 6 female wistar rats. The test material dissolved in corn oil. The volume was made up to 10 ml to achieve the appropriate concentrations. The dosing solution was prepared fresh, prior to dose administration and given in dose concentration 2000mg/kg bw by oral gavage route. 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 provided ad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs. Three rats of first group were dosed with starting dose of 2000 mg/kg body weight and the animals did not show any mortality so another three animals of the same group were dosed with 2000 mg/kg body weight and no mortality was observed. Hence, further dosing was stopped. Body weights were re­corded on day 0 (prior to dosing) 7 and 14. Body weight gain was observed in all surviving animals treated with 2000 mg/kg body weight, during the 14 day observation period, as compared to day 0. At 2000 mg/kg, animal nos. 1, 4, 5 and 6 were observed normal at 30 minutes, 1 hour, on day 1 to 14 post dosing and moderate to mild lethargy was observed at 2, 3 and 4 hours post dosing. Animal nos. 2 and 3 were observed normal at 30 minutes, 1 hour, on day 1 to 14 post dosing and mild lethargy was observed at 2, 3 and 4 hours post dosing. No external and internal gross pathological changes were seen in the animals treated with 2000 mg/kg body weight during terminal sacrifice. Hence, the LD50 value was considered to be >2000 mg/kg body weight, when female wistar rats were treated with the given test chemical orally.