Registration Dossier

Administrative data

Description of key information

Acute Oral Toxicity:

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.

Acute inhalation toxicity:

LC50 of test compound was found be more than 5 mg/L. when Wistar Albino Male and Female rats were inhaled for 4 hours.

Acute dermal toxicity:

LD50 of test compound was found to be more than 2000 mg/kg b.wt (>2000 mg/kg b.wt.) when Wistar albino male and female rats treated dermally.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
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
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
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
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
Preliminary study:
No data
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

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

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.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Data is Klimiach 2 and from study report

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
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
Qualifier:
according to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Principles of method if other than guideline:
Acute Inhalation Toxicity Study of 9,10-Anthracenedione, 1, 4-Diamino-N-N’-Bis (4-C7-17-Branched Alkyl phenyl) Derives in albino rat
GLP compliance:
no
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
Name: 9, 10-Anthracenedione, 1, 4-Diamino-N-N’-Bis (4-C7-17-Branched Alkylphenyl) Derives
- Physical state: Dark blue liquid
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Source : Institute for Industrial Research & Toxicology
Age : 7 to 9 weeks
Sex : Male and Female
Body weight range :200 ±20g
Identification : By cage tag and corresponding colour body marking
No. of animals per dose group :10 (5 males and 5 females)
No. of dose group : Two :Group I – Limit test (5 mg/L) and Group II – Confirmatory test (5 mg/L)
Acclimatization :Twenty healthy albino rats were selected and acclimatized for standard laboratory condition for period of one week in experimental room under veterinary examination.
Randomization : After acclimatization and veterinary examination all the selected rats randomly divided into two groups of five females and five males.
Nutritional conditions :For feeding conventional Laboratory diets may be used with an unlimited supply of drinking water.
HUSBANDRY
Environmental conditions :Air conditioned rooms with 10-15 air changes per hour, temperature between 19-25 0C, relative humidity 30-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.
Accommodation :Groups of five animals of similar sex in polypropylene cages with stainless steel grill top, facilities for food and water bottle, and bedding of clean paddy husk.
Diet : Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India
Water : Community tap water passed through ‘Aqua Guard on line water filter’, was kept in glass bottles, ad-libitum
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: 0.1% tween 80 in Distilled water
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:Nanotek aerosol generator
- Exposure chamber volume: 8 liters
- Method of holding animals in test chamber:For the inhalation purpose the rats were placed in polycarbonate holder tubes positioned radically around exposure chamber, so that only the snouts and nostrils of the animals were exposed to the aerosol.
- Source and rate of air: No data available
- Method of conditioning air:The chamber was maintained at a slightly negative pressure to prevent leakage of the test atmosphere from system, as well as its dilution with outside air.
- System of generating particulates/aerosols:Nanotek aerosol generator
- Method of particle size determination:
- Treatment of exhaust air: The exhaust air was decontaminated by subsequent passage through 1% NaOH solution, silica gel and activated charcoal filters.
- Temperature, humidity, pressure in air chamber: Temperature- 23.29±1.08 - 23.47±1.74, Relative humidity (%): 46.31±3.24 - 47.34±3.56

TEST ATMOSPHERE
- Brief description of analytical method used: No data available
- Samples taken from breathing zone: no

VEHICLE
- Composition of vehicle (if applicable):0.1% tween 80 in Distilled water
- Concentration of test material in vehicle (if applicable): 5 mg/L
- Justification of choice of vehicle: No data available
- Lot/batch no. (if required): No data available
- Purity: No data available

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution: less than 1 micron
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): No data available

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration:No data available
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
4 h
Concentrations:
5 mg/L
No. of animals per sex per dose:
Total: 20
Group I – Limit test (5 mg/L) : 5 males and 5 females
Group II – Confirmatory test (5 mg/L): 5 males and 5 females
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:closely observed for any clinical signs of toxicity at various intervals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days.
- Necropsy of survivors performed: yes
- Other examinations performed: Mortality, clinical signs, body weight and gross pathology were examined.
Key result
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air (nominal)
Based on:
test mat.
Exp. duration:
4 h
Remarks on result:
other: No mortality observed
Mortality:
No mortality was recorded in any of the Wistar albino rats in both limit test and confirmatory test after 4 hours of exposure.
Clinical signs:
other: No clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days.
Body weight:
No significant increase or decrease in body weight on day 7th and 14th were observed when compared with day 0.
Gross pathology:
No gross pathological changes were observed in treated rats.

EXPOSURE ATMOSPHERE DATA 

Parameters

Group-I (5 mg/L)

(limit test)

Group-II (5 mg/L)

(confirmatory test)

Chamber temperature °C (Mean ± S.E.)

23.29±1.08

23.47±1.74

Relative humidity (%)

 (Mean ± S.E.)

46.31±3.24

47.34±3.56

Oxygen content (%)

(Mean ± S.E.)

20.21±1.16

21.34±1.52

 

Mean Body Weight (gm)

S.No.

Groups

BODY WEIGHT (gm)

DAY 0

DAY 7th

% gain or loss

DAY14th

% gain or loss

1.

Group-I (5.0 mg/L)

200.37

205.69

2.65

211.71

5.65

2.

Group-II (5.0 mg/L)

203.11

209.82

3.30

214.24

5.47

CLINICAL SIGNS AND MORTALITY

Group: I Limit test                                                                         Dose: 5.0 mg/L

 

Parameters

Incidence of Clinical Signs Observed after Dosing on

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0/10

 

0

Clinical Signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0         =   No clinical sign (Normal)

+         =   Mild

++        =   Moderate

+++      =   High

++++  =         Severe

CLINICAL SIGNS AND MORTALITY

Group: II Confirmatory test                                                                    Dose: 5.0 mg/L

 

Parameters

Incidence of Clinical Signs Observed after Dosing on

 

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0/10

 

0

Clinical Signs

0

0

0

0

6

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 0         =   No clinical sign (Normal)

+         =   Mild

++        =   Moderate

+++      =   High

++++    =   Severe

SUMMARY OF NECROPSY FINDING

 

S. No.

 

Fate

 

Wistar albino rats

Dose (mg/l)

5.0

(limit test)

5.0

(confirmatory test)

1

Terminal sacrifice

10/10

10/10

2

Found Dead

0/10

0/10

3

Abnormalities detected

0/10

0/10

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: I Limit test                                                                          Dose: 5.0 mg/L       

Animal ID

Fate

Time

Gross Findings

20172-1

TS

Day 14

NAD

20172-2

TS

Day 14

NAD

20172-3

TS

Day 14

NAD

20172-4

TS

Day 14

NAD

20172-5

TS

Day 14

NAD

20172-6

TS

Day 14

NAD

20172-7

TS

Day 14

NAD

20172-8

TS

Day 14

NAD

20172-9

TS

Day 14

NAD

20172-10

TS

Day 14

NAD

Day 0 is the day of exposure

TS=Terminal Sacrifice

NAD=No Abnormality Detected

FD=Found Dead

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: II Confirmatory test                                                                    Dose:5.0 mg/L       

Animal ID

Fate

Time

Gross Findings

20172-11

TS

Day 14

NAD

20172-12

TS

Day 14

NAD

20172-13

TS

Day 14

NAD

20172-14

TS

Day 14

NAD

20172-15

TS

Day 14

NAD

20172-16

TS

Day 14

NAD

20172-17

TS

Day 14

NAD

20172-18

TS

Day 14

NAD

20172-19

TS

Day 14

NAD

20172-20

TS

Day 14

NAD

Day 0 is the day of exposure

TS=Terminal Sacrifice

NAD=No Abnormality Detected

FD=Found Dead

Interpretation of results:
other: Not classified
Conclusions:
Acute lethal Concentration (LC50) of test compound was found be more than 5 mg/L. when Wistar Albino Male and Female rats were inhaled for 4 hours.
Executive summary:

In a acute inhalation toxicity study,Wistar Albino Male and Female rats were exposed with test chemical in the concentration of 5 mg/L in a dynamic nose-only cylindrical chamber built from stainless steel and glass. The chamber had a volume of 8 liters with inner and outer chamber to minimize the fluctuation in concentration and temperature for 4 hours. No mortality was recorded in any of the Wistar albino rats in both limit test and confirmatory test after 4 hours of exposure. No clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days. No significant increase or decrease in body weight on day 7thand 14thwere observed when compared with day 0. No gross pathological changes were observed in treated rats. After 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration. No effects were observed in exposed rats at 5 mg/L. Therefore, Acute lethal Concentration (LC50) of test compound was found be more than 5 mg/L. when Wistar Albino Male and Female rats were inhaled for 4 hours.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
5 000 mg/m³
Quality of whole database:
Data is Klimiach 2 and from study report

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
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
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
Acute Dermal Toxicity Study of test chemical in Wistar albino rats
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Source :Institute for Industrial Research and Toxicology Ghaziabad,
Age: 8 to 10 weeks
Sex: Male and female
Body weight range : 200±20g
Identification : By cage tag and corresponding colour body marking
No. of animals per dose group : 10 (5male & 5 female)
No. of dose groups : Group-I: 2000 mg/kg b.wt (limit test), Group-II: 2000 mg/kg b.wt (confirmatory test)
Acclimatization : The healthy wistar albino rats selected for study acclimatized to standard laboratory condition for period of one week under close Veterinary supervision.
Randomization : After acclimation and Veterinary examination all the animals randomly divided into two groups and each group having five male and five female rats.
Nutritional conditions : Animals were fasted overnight prior to test and food was offered three hours after dosing.
Environmental conditions : Air conditioned rooms with 10-15 air changes per hour, temperature between 22-250C, relative humidity 40-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.
Accommodation : Groups of three animals of same sex in polypropylene cages with stainless steel grill top, facilities for food and water bottle, and bedding of clean paddy husk.
Diet : Pelleted feed supplied by Pranav agro Industries Ltd., B7/6 Ramesh Nagar, Delhi, India
Water : Fresh and clean water filtered through ‘Aqua Guard on line water filter’, was kept in glass bottles Ad libitum


Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Back skin of total body surface area
- % coverage: Approximate 10 percent
- Type of wrap if used: Impervious dressing were used.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, cleaned with lukewarm water wiping
- Time after start of exposure:24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg b.wt
- Concentration (if solution):
- Constant volume or concentration used: yes
- For solids, paste formed: no
Duration of exposure:
:24 hours
Doses:
2000 mg/kg b.wt
No. of animals per sex per dose:
Total: 20
2000 mg/kg b.wt (limit test): 5 male & 5 female
2000 mg/kg b.wt (confirmatory test): 5 male & 5 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Animals were observed for first 4 hours and thereafter for every 1 hrs interval for 24 hrs after dosing and twice a day for 14 days
- Necropsy of survivors performed: yes
- Other examinations performed:Mortality, clinical signs, body weight and gross pathology were examined.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality observed
Mortality:
No mortality at 2000 mg/kg b.wt in wistar albino rats were observed throughout the period of observation14 days.
Clinical signs:
No clinical signs of toxicity were observed in entire the observation period of 14 days.
Body weight:
Normal gain in body weight was observed on day 7th and 14th (post treatment) as compared to day 0th (pre treatment).
Gross pathology:
No significant gross pathological changes related to compound toxicity were observed.

SUMMARY OF BODY WEIGHT (GM

Group

Animal ID

Day 0

Day 7

% Gain/loss

Day 14

% Gain/loss

Group-I

2000 mg/kg b. wt

 

 

 

20172-1

201.3

207.1

2.88

213.4

6.01

20172-2

200.2

205.3

2.54

210.3

5.04

20172-3

200.5

206.4

2.94

213.2

6.33

20172-4

203.4

210.3

3.39

212.4

4.42

20172-5

199.6

206.7

3.56

210.5

5.46

20172-6

206.4

210.4

1.93

215.3

4.31

20172-7

202.7

207.4

2.31

212.1

4.63

20172-8

207.5

212.4

2.36

217.3

4.72

20172-9

203.4

209.3

2.90

212.4

4.42

20172-10

202.1

205.5

1.68

212.4

5.09

Group-II

2000 mg/kg b. wt

20172-11

198.4

209.2

5.44

208.1

4.88

20172-12

205.5

211.3

2.82

216.6

5.40

20172-13

206.4

212.2

2.81

217.4

5.32

20172-14

202.4

207.4

2.47

213.4

5.43

20172-15

203.5

207.1

1.76

213.1

4.71

20172-16

207.7

214.3

3.17

217.4

4.67

20172-17

200.4

207.3

3.44

209.2

4.39

20172-18

197.4

202.7

2.68

210.2

6.48

20172-19

205.3

210.4

2.48

216.1

5.26

20172-20

206.6

211.4

2.32

218.4

5.71

CLINICAL SIGNS AND MORTALITY

Group: I Limit test                                                 Dose: 2000 mg/kg b.wt

 

Parameters

Incidence of Clinical Signs Observed after Dosing on

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/10

0

Clinical Signs- Local

 

Redness

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Pain

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Swelling

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Systemic signs

Clinical signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

-          =Observed after 24 hrs

0        = No clinical signs

+        = Mild

++      = Moderate

+++    = High

++++  = Severe

CLINICAL SIGNS AND MORTALITY

Group: II Confirmatory test                                               2000 mg/kg b.wt

 

Parameters

Incidence of Clinical Signs Observed after Dosing on

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/10

0

Clinical Signs- Local

 

Redness

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Pain

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Swelling

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Systemic signs

Clinical signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

-          =Observed after 24 hrs

0        = No clinical signs

+        = Mild

++      = Moderate

+++    = High

++++  = Severe

SUMMARY OF NECROPSY FINDINGS

S. No.

 

Fate

 

Wistar albino rats

Dose (mg/kg b. wt)

2000

(limit test)

2000

(confirmatory test)

1

Terminal sacrifice

10/10

10/10

2

Found Dead

0/10

0/10

3

Abnormalities detected

0/10

0/10

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group-I (limit test)                                                                         2000 mg/kg b.wt.

Animal ID

Fate

Time

Gross Findings

20172-1

TS

Day 14

NAD

20172-2

TS

Day 14

NAD

20172-3

TS

Day 14

NAD

20172-4

TS

Day 14

NAD

20172-5

TS

Day 14

NAD

20172-6

TS

Day 14

NAD

20172-7

TS

Day 14

NAD

20172-8

TS

Day 14

NAD

20172-9

TS

Day 14

NAD

20172-10

TS

Day 14

NAD

Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

INDIVIDUAL ANIMAL FATE & NECROPSY FINDING

Group: II(confirmatory test)                                                     Dose: 2000 mg/kg b.wt.

Animal ID

Fate

Time

Gross Findings

20172-11

TS

Day 14

NAD

20172-12

TS

Day 14

NAD

20172-13

TS

Day 14

NAD

20172-14

TS

Day 14

NAD

20172-15

TS

Day 14

NAD

20172-16

TS

Day 14

NAD

20172-17

TS

Day 14

NAD

20172-18

TS

Day 14

NAD

20172-19

TS

Day 14

NAD

20172-20

TS

Day 14

NAD

 Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

FD-Found dead

FD-Found dead

Interpretation of results:
other: Not classified
Conclusions:
LD50 of test compound was found to be more than 2000 mg/kg b.wt. (>2000 mg/kg b.wt.) when Wistar albino male and female rats.treated dermally.
Executive summary:

In a acute dermal toxicity study, Wistar albino male and female rats treated with test chemical dermally at 2000 mg/kg b.wt applied uniformly over approximate 10 percent back skin of total body surface area for 24 hrs with an impervious dressing secured in place with an adhesive tape. The animals were then housed individually in cages with a collar around the neck in order to avoid the ingestion of the test compound. After 24 hours, the dressing was removed and the site of application was cleaned with lukewarm water wiping. No mortality at 2000 mg/kg b.wt in wistar albino rats were observed throughout the period of observation14 days. No clinical signs of toxicity were observed in entire the observation period of 14 days. Normal gain in body weight was observed on day 7th and 14th (post treatment) as compared to day 0th (pre treatment). No significant gross pathological changes related to compound toxicity were observed. Therefore, LD50 of test compound was found to be more than 2000 mg/kg b.wt. (>2000 mg/kg b.wt.) when Wistar albino male and female rats treated dermally.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Data is Klimiach 2 and from study report

Additional information

Acute Oral toxicity:

In a study, test chemical has been reviewed for acute oral toxicity to a greater or lesser extent. Study based on in vivo experiments in rodents, i.e. most commonly in rats.

In a experimental 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 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.

Thus, based on the above study on test chemical, it can be concluded that LD50 value is greater 2000 mg/kg bw for acute oral toxicity. Thus comparing this value with the criteria of CLP regulation, test chemical can be Not classified for acute oral toxicity.

Acute inhalation toxicity:

In a study, test chemical has been reviewed for acute inhalation toxicity to a greater or lesser extent. Study based on in vivo experiments in rodents, i.e. most commonly in rats.

In a experimental study, Wistar Albino Male and Female rats were exposed with test chemical in the concentration of 5 mg/L in a dynamic nose-only cylindrical chamber built from stainless steel and glass. The chamber had a volume of 8 liters with inner and outer chamber to minimize the fluctuation in concentration and temperature for 4 hours. No mortality was recorded in any of the Wistar albino rats in both limit test and confirmatory test after 4 hours of exposure. No clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days. No significant increase or decrease in body weight on day 7thand 14thwere observed when compared with day 0. No gross pathological changes were observed in treated rats. After 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration. No effects were observed in exposed rats at 5 mg/L. Therefore, Acute lethal Concentration (LC50) of test compound was found be more than 5 mg/L. when Wistar Albino Male and Female rats were inhaled for 4 hours.

Thus, based on the above study on test chemical, it can be concluded that LD50 value is greater 5 mg/L. for acute inhalation toxicity. Thus comparing this value with the criteria of CLP regulation, test chemical can be Not classified for acute inhalation toxicity.

Acute dermal toxicity:

In a study, test chemical has been reviewed for acute dermal toxicity to a greater or lesser extent. Study based on in vivo experiments in rodents, i.e. most commonly in rats.

In a experimental study, Wistar albino male and female rats treated with test chemical dermally at 2000 mg/kg b. wt applied uniformly over approximate 10 percent back skin of total body surface area for 24 hrs with an impervious dressing secured in place with an adhesive tape. The animals were then housed individually in cages with a collar around the neck in order to avoid the ingestion of the test compound. After 24 hours, the dressing was removed and the site of application was cleaned with lukewarm water wiping. No mortality at 2000 mg/kg b.wt in wistar albino rats was observed throughout the period of observation14 days. No clinical signs of toxicity were observed in entire the observation period of 14 days. Normal gain in body weight was observed on day 7th and 14th (post treatment) as compared to day 0th (pre treatment). No significant gross pathological changes related to compound toxicity were observed. Therefore, LD50 of test compound was found to be more than 2000 mg/kg b.wt (>2000 mg/kg b.wt.) when Wistar albino male and female rats treated dermally.

Thus, based on the above study on test chemical, it can be concluded that LD50 value is greater 2000 mg/kg bw for acute dermal toxicity. Thus comparing this value with the criteria of CLP regulation, test chemical can be Not classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above study on test chemical, it can be concluded that LD50 value is greater 5 mg/L and 2000 mg/kg bw for acute oral, inhalation and dermal toxicity. Thus comparing this value with the criteria of CLP regulation, test chemical can be Not classified for acute oral, inhalation and dermal toxicity.