Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
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:
2017
Report Date:
2017

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:
Acute Oral Toxicity of test chemical in Rats.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Name - 9,10-Anthracenedione, 1,4-diamino-, N,N'-bis(4-C7-17-branched alkylphenyl) derivs.
Consistency: Liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: approximately 8 to 12 weeks old
- Weight at study initiation: Body weight range was 199.8 to 211.7 grams.
Body weights at the start : Mean: 204.54 g (= 100 %), Minimum : 199.8 g (- 2.32 %), Maximum : 211.7 g (+ 3.50 %)
- Fasting period before study: approximately 16 hours or more
- Housing: The rats were housed in polycarbonate cages.
- Diet (e.g. ad libitum): Rodent feed supplied ad libitum.
- Water (e.g. ad libitum):Water was provided ad libitum from individual bottles attached to the cages.
- Acclimation period: at least 5 days prior to administration

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.1 to 21.9 °C
- Humidity (%): 56.3% to 59.8%
- Air changes (per hr): at least 10 to 15 air changes/hour of 100% fresh air that had been passed through the HEPA filters.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room

IN-LIFE DATES: From: 08-06-2017 To:02-08-2017

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 300 mg/kg and 2000 mg/kg
- Amount of vehicle (if gavage):10 ml/kg body weight
- Justification for choice of vehicle: Given test solution is soluble in corn oil.
- Lot/batch no. (if required): No data
- Purity: No data

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg

DOSAGE PREPARATION (if unusual): The test item was prepared by dilution of the test item in corn oil to obtain 30 mg/ml strength of solution. The formulation was prepared fresh on the day of dosing. The test item was administered in the dose volume of 10 ml/kg body weight.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: 300 mg/kg
Doses:
300 mg/kg and 2000 mg/kg
No. of animals per sex per dose:
Total - 12 (Females)
Step I (300 mg/kg) - 3
Step II (300 mg/kg) - 3
Step I (2000 mg/kg) - 3
Step II (2000 mg/kg) - 3
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations :Animals were observed for clinical signs, mortality and morbidity, until sacrifice.The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time.
- Weighing: Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, were performed.
Statistics:
No data

Results and discussion

Preliminary study:
No data
Effect levelsopen allclose all
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 50% nortality was obserbed
Mortality:
No mortality was observed at 2000 mg/kg.
Clinical signs:
Group I Step I and II : At 300 mg/kg There was no signs of toxicity on day 2 after the dosing; coloured faces and diarrhoea with onset from 4 hours to day 1 after the dosing was observed.
Group II Step I and II : At 2000 mg/kg There was no signs of toxicity on day 3 after the dosing; coloured faces with onset on day 1 after the dosing was observed.

Body weight:
Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.27% and 13.49% respectively.
Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.98% and 11.96% respectively.
Group II Step I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.73% and 11.36% respectively.
Group II Step II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.90% and 13.15% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.
Other findings:
No data

Any other information on results incl. tables

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

Group I :   

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

300

Test item coloured faces

3

1,2

3

Day 1

4 hrs. - Day 1

0/3

Diarrhoea

1

3

4 hrs. - 6 hrs.

Group I :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

II

300

Test item coloured faces

3

4

5

6

4 hrs. - Day 1

Day 1

6 hrs. - Day 1

0/3

Diarrhoea

2

4

6

4 hrs. - 6 hrs.

6 hrs.

 

Group II :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

2000

Test item coloured faces

3

7,8,9

Day 1 - Day 2

0/3

 

Group II :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

II

2000

Test item coloured faces

3

10,11,12

Day 1 - Day 2

0/3

Table No.II

Mean Body Weight and Percent Body Weight Gain (g)

Group I :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

300

Mean

201.83

214.50

6.27

229.07

6.79

13.49

± SD

1.78

3.47

0.82

4.38

0.35

1.23

 

Group I :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

II

300

Mean

204.17

216.37

5.98

228.57

5.64

11.96

± SD

4.01

4.13

0.71

3.81

0.72

0.33

 

 Group II :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

2000

Mean

208.17

220.07

5.73

231.77

5.33

11.36

± SD

4.88

4.18

0.83

2.35

1.08

1.87

 

 Group II :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

II

2000

Mean

204.00

216.03

5.90

230.83

6.85

13.15

± SD

2.36

2.30

0.19

3.73

0.62

0.52

Table No.III

Summary of Gross Pathological Findings

Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

300

1 - 3

TS

No abnormality detected

 

        Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

300

4 - 6

TS

No abnormality detected

 

                    Group II :

 

Step No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

7 - 9

TS

No abnormality detected

 

                    Group II :

 

Step No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

2000

10 - 12

TS

No abnormality detected

                     TS = Terminal Sacrifice

 

Appendix No.I

Individual Animal -Clinical Signs of Toxicity and Mortality

 Group I :   

 

Step

No.

Dose

mg/kg

Total Number of

Animals

Observed Signs

Animal Nos.

Period of signs

in days

From - to

Mortality

I

300

3

Test item coloured faces

1

Day 1

0

Test item coloured faces

2

Day 1

0

Test item coloured faces

3

4 hrs. - Day 1

0

Diarrhoea 

3

4 hrs. - 6 hrs.

 

  Group I :

Step

No.

Dose

mg/kg

Total Number of

Animals

Observed Signs

Animal Nos.

Period of signs

in days

From - to

Mortality

II

300

3

Test item coloured faces

4

4 hrs. - Day 1

0

Diarrhoea 

4

4 hrs. - 6 hrs.

Test item coloured faces

5

Day 1

0

Test item coloured faces

6

6 hrs. - Day 1

0

Diarrhoea 

6

6 hrs.

 

  Group II :

Step

No.

Dose

mg/kg

Total Number of

Animals

Observed Signs

Animal Nos.

Period of signs

in days

From - to

Mortality

I

2000

3

Test item coloured faces

7

Day 1 - Day 2

0

Test item coloured faces

8

Day 1 - Day 2

0

Test item coloured faces

9

Day 1 - Day 2

0

 

  Group II :

Step

No.

Dose

mg/kg

Total Number of

Animals

Observed Signs

Animal Nos.

Period of signs

in days

From - to

Mortality

II

2000

3

Test item coloured faces

10

Day 1 - Day 2

0

Test item coloured faces

11

Day 1 - Day 2

0

Test item coloured faces

12

Day 1 - Day 2

0

 

 

Appendix No.II

Individual Animal - Body Weight and Percent Body Weight Gain (g)

Group : I                     Step I :                                Dose  : 300 mg/kg body weight

Animal No.

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

1

199.8

210.7

5.46

224.4

6.50

12.31

2

203.1

217.5

7.09

233.1

7.17

14.77

3

202.6

215.3

6.27

229.7

6.69

13.38

 

Group : I                     Step II :                              Dose  : 300 mg/kg body weight

Animal No.

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

4

208.6

220.2

5.56

232.8

5.72

11.60

5

203.1

216.9

6.79

227.5

4.89

12.01

6

200.8

212.0

5.58

225.4

6.32

12.25

 

Group : II                    Step I :                                Dose  : 2000 mg/kg body weight

Animal No.

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

7

210.2

223.1

6.14

234.1

4.93

11.37

8

211.7

221.8

4.77

231.8

4.51

9.49

9

202.6

215.3

6.27

229.4

6.55

13.23

 

Group : II                    Step II :                               Dose  : 2000 mg/kg body weight

Animal No.

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

10

201.8

213.7

5.90

227.4

6.41

12.69

11

206.5

218.3

5.71

234.8

7.56

13.70

12

203.7

216.1

6.09

230.3

6.57

13.06

 

Appendix No.III

Individual Animal - Gross Pathological Findings

Group : I

Step I :

Dose  : 300 mg/kg body weight

Animal No.

Fate

Gross Pathological Findings

1

TS

No abnormality detected

2

TS

No abnormality detected

3

TS

No abnormality detected

 

Group : I

Step II :

Dose  : 300 mg/kg body weight

Animal No.

Fate

Gross Pathological Findings

4

TS

No abnormality detected

5

TS

No abnormality detected

6

TS

No abnormality detected

 

Group : II

Step I :

Dose  : 2000 mg/kg body weight

Animal No.

Fate

Gross Pathological Findings

7

TS

No abnormality detected

8

TS

No abnormality detected

9

TS

No abnormality detected

 

Group : II

Step II :

Dose  : 2000 mg/kg body weight

Animal No.

Fate

Gross Pathological Findings

10

TS

No abnormality detected

11

TS

No abnormality detected

12

TS

No abnormality detected

 

TS = Terminal sacrifice

Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
The LD50 value of the test compound was found to be more than 2000 mg/kg bw (>2000 mg/kg bw) when Sprague Dawley female rats treated via oral gavage route.
Executive summary:

In a acute oral toxicity study, 12 Sprague Dawley female rats treated with test chemical by oral gavage route at the concentration of 300 and 2000 mg/kg bw in 4 different steps.The dose of 300 mg/kg of test item was prepared by dilution of the test item in corn oil to obtain 30 mg/ml strength of solution. The formulation was prepared fresh on the day of dosing. The test item was administered in the dose volume of 10 ml/kg body weight. The dose of 2000 mg/kg of test item was administered undiluted . Female rats of the age of approximately 8 to 12 weeks old were used and its weight were within ± 20% of the mean weight of any animal used for dosing. Body weight range was 199.8 to 211.7 grams. The rats were housed in polycarbonate cages.The animal room was independently provided with at least 10 to 15 air changes per hour of 100% fresh air that had been passed through the HEPA filters. Room temperature was maintained at 20.1 to 21.9° C and room humidity was maintained at 56.3% to 59.8%.An artificial light and dark cycle of 12 hours each was provided to the room.Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.The single dose of test item was administered to fasted rats (approximately 16 hours or more) by oral intubation, using a ball-tipped intubation needle fitted onto a syringe of appropriate size.

Initially, 3 female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in test item coloured faces and diarrhoea with onset from 4 hours to day 1 after the dosing and no mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, 3 female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg resulted in test item coloured faces and diarrhoea with onset from 4 hours to day 1 after the dosing and no mortality after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional 3 female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in test item coloured faces with onset on day 1 after the dosing and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, additional 3 female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in test item coloured faces with onset on day 1 after the dosing and no mortality after the dosing. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups. Therefore, LD50 value of the test chemical was found to be more than 2000 mg/kg bw (>2000 mg/kg bw) when Sprague Dawley female rats treated via oral gavage route.