9,10-Anthracenedione, 1,4-diamino-, N,N'-bis(4-C7-17-branched alkylphenyl) derivs.

Acute Toxicity: oral

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

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

Test material information

Constituent 1

Reference substance name:

9,10-Anthracenedione, 1,4-diamino-, N,N'-bis(4-C7-17-branched alkylphenyl) derivs.

EC Number:

308-067-0

EC Name:

9,10-Anthracenedione, 1,4-diamino-, N,N'-bis(4-C7-17-branched alkylphenyl) derivs.

Cas Number:

97862-23-2

Molecular formula:

C48H62N2O2

IUPAC Name:

9,10-Anthracenedione, 1,4-diamino-, N,N'-bis(4-C7-17-branched alkylphenyl) derivs.

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 or test system 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 levels

Effect levels 1

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

Effect levels 2

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.