Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on study conducted on rats for the test chemical.The study concluded that the LD50 value is >2000 mg/kg bw, for acute oral toxicity.Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity. 

Acute Inhalation toxicity: 

The study need not be conducted because the exposure of the test chemical via inhalation is not likely owing its low vapour pressure and/or the possibility of exposure to aerosols, particles or droplets of inhalable size. The estimated vapour pressure of test substance at 25 Deg C was 0.0000000809 Pa. Hence, this endpoint was considered for waiver.

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on experimental study conducted on rats for the test chemical. The study concluded that the LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

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 with acceptable restrictions
Justification for type of information:
Data is from experimental study.
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
The study was conducted to findout mortality, clinical signs, necropsy finding and histopathological changes in wistar albino rats by using the given test chemical.
GLP compliance:
not specified
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Institute for Industrial Research & Toxicology
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 7 to 9 weeks
- Weight at study initiation: 200±20g
- Identification:By cage tag and corresponding colour body marking
- Fasting period before study: Fasted overnight prior to treatment. Food was offered three hours after dosing.
- Housing: Groups of three 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 (e.g. ad libitum): Pelleted feed
- Water (e.g. ad libitum): Aqua Guard filter water was kept in PVC bottles, ad libitum
- Acclimation period: One week in experimental room after veterinary examination.
- Randomization:After acclimation and veterinary examination randomly selected in groups of three females.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature between 22-25°C
- Humidity (%): relative humidity 40-60%
- Air changes (per hr): Air conditioned rooms with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 2000 mg/kg bw
- Amount of vehicle (if gavage): 10ml/kg bw

DOSAGE PREPARATION (if unusual): Dose preparation of the test article was done freshly, few minutes prior to dosing. Test substance was dissolved in distilled water to obtain final concentration of 200 mg/ml.
Doses:
Two + one vehicle control
Group I: Dist. water, 10ml/kg body wt.
Group II : 2000 mg/kg body wt.
Group III : 2000 mg/kg body wt.
No. of animals per sex per dose:
Three (3 females)/step
Control animals:
yes
Remarks:
Group I: Dist. water, 10ml/kg body wt.
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality: The test compound was administered at different dose level in wistar albino rats observed for mortality at the time interval of 30 minutes, 1hr, 2hr, 4 hr, and 6hr time interval on the day of test compound administration and thereafter twice a day for 14 days.
Body weight: The body weight of all the animals was observed weekly on day 0 (pre treatment), 7th and 14th (post treatment).
Clinical Signs: The treated animals were closely observed for clinical signs of intoxication, first 4 hours and every 1 hrs interval for 24 hrs after dosing and thereafter twice a day for 14 days. All the rats were observed at least twice daily to observe any clinical signs or behavioral changes. These observations included changes in skin and fur, in the eyes and mucous membranes, respiratory, circulatory, central nervous and autonomic nervous systems, somatomotor activity and behavioral changes. The following clinical signs were observed in female mice to characterize the various systemic studies: Salivation, lacrimation, pale mucous membrane, diarrheal feaces, hunched posture, scratching, polyuria, hypoactivity etc.
- Necropsy of survivors performed: yes, necropsy was carried out on all the animals which died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes.
Statistics:
not specified
Preliminary study:
not specified
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality recorded in any of the Wistar albino rats after administration of the test compound during the entire observation period (14 days).
Clinical signs:
The test compound did not produce any clinical signs of toxicity at the tested dose level throughout the period of observation.
Body weight:
Wistar albino rats treated with the test compound at the dose level of 2000 mg/kg b.wt. showed significant normal gain in body weight on day 7th and 14th (post treatment) as compared to control group.
Gross pathology:
NECROPSY FINDING
EXTERNAL
i.Skin- Skin and hair coat was observed wet.
ii.All external orifices- Normal
B. INTERNAL
i. Subcutaneous- No change was observed.
ii. Superficial and deep lymph nodes- No change in mesenteric lymph node.
ABDOMINAL CAVITY
i.Opening and general examination- In the abdominal cavity all the organs were present in normal position.
ii.Spleen- Normal upto highest tested dose level 2000 mg/kg b.wt.
iii.Digestive system- No gross changes were observed in stomach and intestine upto highest tested dose level 2000 mg/kg b.wt.
iv.Liver and biliary ducts- No gross pathological changes were observed
v.Excretory system- No gross pathological changes were observed upto highest tested dose level 2000 mg/kg b.wt.
vi.Adrenal- Observed normal.
vii.Male/female genital organs – Showed normal colour, consistency and no inflammatory changes upto highest tested dose level 2000 mg/kg b.wt.
2. THORACIC CAVITY
i.Opening and general examination- Thoracic cavity was found to be normal without any fluid, mucous or blood etc.
ii.Lungs- observed normal.
iii.Heart- No changes were observed in color and consistency. Heart found normal upto highest tested dose level 2000 mg/kg b.wt.
iv.Thyroid- Normal in shape, size and surface upto highest tested dose level 2000 mg/kg b.wt.
3. CRANIAL CAVITY
Brain- Normal in shape and size.
Other findings:
not specified

TABLE – 2

SUMMARY OF BODY WEIGHT (GM)

Group

Day 0

Day 7

% Gain/loss

Day 14

% Gain/loss

Group-I distilled water 10ml/kg

202.40

208.90

3.21

217.40

7.41

Group-II

2000 mg/kg b. wt

203.70

209.2

2.70

219.9

7.95

Group-III

2000 mg/kg b. wt

201.90

208.30

3.16

216.70

7.33


TABLE – 3

CLINICAL SIGNS AND MORTALITY

Group: I (Vehicle Control)                                                           Dose: 10 ml/kg b.wt

                                           

FEMALE RATS 

Parameters

Incidence of clinical signs observed after dosing

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

 

0/3

 

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)

+         =   Clinical Sign

 


TABLE – 3 (Contd.)

CLINICAL SIGNS AND MORTALITY

Group: II                                                                                           Dose: 2000 mg/kg b. wt.

 

FEMALE RATS

 

Parameters

Incidence of clinical signs observed after dosing

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

 

0/3

Clinical Signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

  0           =   Normal

+            =    Mild

++           =   Moderate

+++         =   High

 ++++     =   Severe


TABLE – 3 (Contd.)

CLINICAL SIGNS AND MORTALITY

Group: III                                                                                          Dose: 2000 mg/kg b.wt

                                            

FEMALE RATS 

 

Parameters

Incidence of clinical signs observed after dosing

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

 

0/3

Clinical Signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 0            =   Normal

+            =    Mild

++           =   Moderate

+++         =   High

 ++++     =   Severe


TABLE – 4

SUMMARY OF NECROPSY FINDINGS

S. No.

 

Fate

 

Wistar albino rats

Dose (mg/kg b. wt)

Distilled water (10 ml/kg)

 

2000

 

 

2000

1

Terminal sacrifice

3/3

3/3

3/3

2

Found Dead

0/3

0/3

0/3

3

Abnormalities detected

NAD

NAD

NAD

NAD - No abnormality recorded


TABLE - 5

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: I (Vehicle Control)                                                           Dose: 10ml/kg b.wt

                                                                                                                    

FEMALE RATS

 

Animal ID

Fate

Time

Gross Findings

20164-1

TS

Day 14

NAD

20164-2

TS

Day 14

NAD

20164-3

TS

Day 14

NAD

 

Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

 

 

TABLE – 5 Contd……..

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: II                                                                                            Dose: 2000 mg/kg b.wt.                  

FEMALE RATS

 

Animal ID

Fate

Time

Gross Findings

20164-4

TS

Day 14

NAD

20164-5

TS

Day 14

NAD

20164-6

TS

Day 14

NAD

 

Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

FD- Found Dead

 

TABLE – 5 contd……….

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: III                                                                                          Dose: 2000 mg/kg b.wt.      

                                                     

                                                           FEMALE RATS 

Animal ID

Fate

Time

Gross Findings

20164-7

TS

Day 14

NAD

20164-8

TS

Day 14

NAD

20164-9

TS

Day 14

NAD

 

Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

FD- Found Dead 

Interpretation of results:
other: Not classified
Conclusions:
From the results obtained from present investigation, the LD50 value was considered to be >2000 mg/kg bw. Thus, it can be concluded that the test chemical is non toxic to Wistar albino rats. CLP classification "Not classified"
Executive summary:

The acute oral toxicity study was conducted under the OECD Guideline-423 for testing of chemicals in wistar albino rats. The healthy wistar albino rats of body weight 200±20 gm were selected for study after acclimatization to standard laboratory condition and divided into test compound and vehicle control group each having three animals. The study was conducted stepwise as follow:

Starting dose 2000 mg/kg body weight: Step-I -  The test compound was mixed with distilled water and administered orally at the dose level of 2000 mg/kg body weight (dose volume 10ml/kg) to three female rats. However; vehicle control group treated with the distilled water at the dose level of 10 ml/kg b.wt.  The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days.  The necropsy was performed on all animals at the termination of the study.

The test compound did not elicit any clinical signs of intoxication throughout the period of observation at the tested dose level 2000 mg/kg body weight. Furthermore, no mortality was recorded at test dose level. Vehicle control group of animals were also free from any mortality and clinical signs. The necropsy finding did not reveal any gross pathological changes at the tested dose level. Furthermore, no gross pathological change was observed in vehicle control group.

Step -II: After 72 hrs, the result of step-I was confirmed by administration of same dose level (2000 mg/kg. b.wt) of test compound in additional three animals of same sex (OECD-423 guidelines) under same test condition.

The test compound administered at the dose level of 2000 mg/kg b.wt did not produce any mortality and clinical sign of intoxication throughout the observation period of 14 days. Body weight of each rat was also recorded on day 7th and 14th showed normal gain as compared to control group. Necropsy was conducted at the end of the study (15th day) on all the animals which did not reveal any gross pathological changes.

From the results obtained from present investigation, the LD50 value was considered to be >2000 mg/kg bw. Thus,  it can be concluded that the test chemical is non toxic to Wistar albino rats. CLP classification "Not classified"

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

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
other justification
Justification for data waiving:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
Endpoint conclusion
Quality of whole database:
waiver

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 with acceptable restrictions
Justification for type of information:
Data is from experimental report.
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
The study was conducted to find out the mortality, clinical sign of toxicity and histopathological effect of the given test chemical at different dose level in wistar albino rats.
GLP compliance:
not specified
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Institute for Industrial Research and Toxicology Ghaziabad
- Age at study initiation: 8 to 10 weeks
- Weight at study initiation: 200±20g
- Fasting period before study: Animals were fasted overnight prior to test and food was offered three hours after dosing.
- Housing: 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.
- Identification: By cage tag and corresponding colour body marking
- Diet (e.g. ad libitum): Pelleted feed
- Water (e.g. ad libitum): Fresh and clean water filtered through ‘Aqua Guard on line water filter’, was kept in glass bottles Ad libitum
- Acclimation period: 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.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): temperature between 22-25°C
- Humidity (%): relative humidity 40-60%
- Air changes (per hr): Air conditioned rooms with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark
Type of coverage:
occlusive
Vehicle:
water
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: The test compound was held in contact with the skin 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.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The dressing was removed and the site of application was cleaned with lukewarm water wiping the test compound.
- Time after start of exposure: 24 hours
Duration of exposure:
24 hours
Doses:
Group-I: 2000 mg/kg b.wt (limit test)
Group-II: 2000 mg/kg b.wt (confirmatory test)
No. of animals per sex per dose:
10 (5male & 5 female)
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality - All the animals were observed for mortality at 30 minutes time interval for first six hours on the day of test compound administration and thereafter twice a day for 14 days.
Body weight: The body weight of all the animals was observed weekly on day 0 (pre treatment), 7th and 14th (post treatment).
Clinical signs - The treated animals were closely observed for clinical signs of intoxication, first 4 hours and thereafter for every 1 hrs interval for 24 hrs after dosing and twice a day for 14 days. All the rats were observed at least twice daily with the purpose of recording any symptoms of ill-health or behavioral changes. These observations included changes in skin and fur in the eyes and mucous membranes, respiratory, circulatory, central nervous and autonomic nervous systems, somatomotor activity and behavioral changes. The following clinical signs were observed in female rats to characterize with erythema, hypersensitivity, edema etc.
- Necropsy of survivors performed: yes, necropsy was carried out on all the animals that died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes.
Statistics:
not specified
Preliminary study:
not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
The test compound when applied dermally at the dose level of 2000 mg/kg b.wt. on Wistar albino rats did not produce any mortality during the observation period of 14 days.
Clinical signs:
The Wistar albino rats treated with the test compound were free from any systematic and local clinical signs when observed after 24th hours of patch removal.
Body weight:
The body weight of all the animals was recorded on day 0 (pre treatment) and then 7th and 14th (post treatment) and it has been observed that there was no significant increase and decrease in the body weight. However, normally increase in the body weight was recorded on day 14th.
Gross pathology:
NECROPSY FINDING
EXTERNAL
i.Skin- Skin and hair coat was observed wet.
ii.All external orifices- Normal
B. INTERNAL
i. Subcutaneous- No changes were observed.
ii. Superficial and deep lymph nodes- No change in mesenteric lymph node.
ABDOMINAL CAVITY
i.Opening and general examination- In the abdominal cavity all the organs were present in normal position.
ii.Spleen- No changes were recorded.
iii.Digestive system- No gross changes were observed in stomach and intestine.
iv.Liver and biliary ducts- No gross pathological changes were observed
v.Excretory system- No gross pathological changes were observed.
vi.Adrenal- Observed normal.
vii.Male/female genital organs – Showed normal colour, consistency and no inflammatory changes.
2. THORACIC CAVITY
i.Opening and general examination- Thoracic cavity was found to be normal without any fluid, mucous or blood etc.
ii.Lungs- No changes were recorded.
iii.Heart- No changes were observed in color and consistency. Heart found normal.
iv.Thyroid- Normal in shape, size and surface.
3. CRANIAL CAVITY
Brain- Normal in shape and size.
Other findings:
not specified

TABLE – 2

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

 

 

 

20164-1

203.15

206.12

1.46

210.90

3.80

20164-2

201.91

204.95

1.50

208.99

3.50

20164-3

201.12

213.47

2.08

217.50

4.0

20164-4

211.12

215.91

2.26

219.98

3.82

20164-5

200.19

204.93

2.36

208.13

3.96

20164-6

198.99

203.01

2.02

207.0

4.02

20164-7

203.40

206.50

2.15

210.75

3.61

20164-8

207.6

212.12

1.96

215.36

4.00

20164-9

204.91

207.98

1.47

21.03

3.47

20164-10

207.11

210.19

1.48

214.50

3.56

Group-II

2000 mg/kg b. wt

20164-11

200.32

203.94

1.80

207.92

3.79

20164-12

201.91

204.99

1.52

208.62

3.32

20164-13

207.21

211.29

1.96

215.92

4.20

20164-14

205.14

209.31

2.03

213.34

3.99

20164-15

211.20

215.93

2.23

219.56

3.99

20164-16

212.12

216.18

1.91

220.38

3.89

20164-17

203.12

207.45

2.13

211.76

4.25

20164-18

200.19

204.91

2.35

218.87

4.33

20164-19

199.80

209.99

2.59

208.77

4.48

20164-20

208.11

212.09

1.91

215.74

3.68

TABLE – 3

CLINICAL SIGNS AND MORTALITY

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

 

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

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

                                                                                                                                                          

-          =Observed after 24 hrs

0        = No clinical signs

+        = Mild

++      = Moderate

+++    = High

++++  = Severe

TABLE – 3 Contd…………….

CLINICAL SIGNS AND MORTALITY

Group: II Confirmatory 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

TABLE – 4

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

TABLE – 5

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

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

Animal ID

Fate

Time

Gross Findings

20164-1

TS

Day 14

NAD

20164-2

TS

Day 14

NAD

20164-3

TS

Day 14

NAD

20164-4

TS

Day 14

NAD

20164-5

TS

Day 14

NAD

20164-6

TS

Day 14

NAD

20164-7

TS

Day 14

NAD

20164-8

TS

Day 14

NAD

20164-9

TS

Day 14

NAD

20164-10

TS

Day 14

NAD

Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

FD-Found dead

 TABLE-5 Contd………..

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

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

Animal ID

Fate

Time

Gross Findings

20164-11

TS

Day 14

NAD

20164-12

TS

Day 14

NAD

20164-13

TS

Day 14

NAD

20164-14

TS

Day 14

NAD

20164-15

TS

Day 14

NAD

20164-16

TS

Day 14

NAD

20164-17

TS

Day 14

NAD

20164-18

TS

Day 14

NAD

20164-19

TS

Day 14

NAD

20164-20

TS

Day 14

NAD

 Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

FD-Found dead

Interpretation of results:
other: Not classified
Conclusions:
From the results obtained from present investigation, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when male/female Wistar albino rats were treated with the given test chemical by dermal application. Thus, it can be concluded that the test compound is non toxic to Wistar albino rats as per the criteria of CLP regulation.
Executive summary:

The acute dermal toxicity study was conducted according to OECD guideline 402 for testing of chemicals on Wistar albino rats. The summary of the study was as follows -

LIMIT TEST (2000 mg/kg b.wt): Ten healthy wistar albino rats of both sex (ranging b.wt 200±20 gm) selected for study after acclimatization. Approximate 10 percent back skin of total body surface area was prepared 24 hrs prior to application of test compound. The test compound was applied dermally at the dose level of 2000 mg/kg b.wt for each animal. The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days. The body weight of each rat was observed on day 0 (pre treatment), 7th and 14th (post treatment). The necropsy was performed on all animals at termination of the study.

The test compound applied at the dose level of 2000 mg/kg b.wt in Wistar albino rats did not show any mortality as well as clinical signs of toxicity throughout the observation period of 14 days.  Necropsy finding did not reveal any gross pathological changes under test condition.

CONFIRMATORY TEST: After 72 hrs, a confirmatory test was conducted in same species of animals to confirm the results obtained from limit test (OECD-402 guidelines). Ten healthy Wistar albino rats of both sex (ranging b.wt 200±20 gm) selected for study after acclimatization. Approximate 10 percent back skin of total body surface area was prepared 24 hrs prior to application of test compound. The test chemical was applied dermally at the dose level of 2000 mg/kg b.wt for each animal. The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days. The body weight of each rat was observed on day 0 (pre treatment), 7th and 14th (post treatment). The necropsy was performed on all animals which was died during the study or were sacrificed at termination of the study.

No mortality was recorded in Wistar albino rats after administration of test compound at the dose level of 2000 mg/kg b.wt throughout the period of observation (14 days). The test compound did not elicit any clinical signs of toxicity during the observation period. No skin reaction was observed after 24th hrs of patch removal. The body weight of each animal recorded on day 7th and 14th showed normal increase as compared to day 0 (pre treatment).

From the results obtained from present investigation, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when male/female Wistar albino rats were treated with the given test chemical by dermal application. Thus, it can be concluded that the test compound is non toxic to Wistar albino rats as per the criteria of CLP regulation.

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

Additional information

Acute oral toxicity:

The acute oral toxicity study was conducted under the OECD Guideline-423 for testing of chemicals in wistar albino rats. The healthy wistar albino rats of body weight 200±20 gm were selected for study after acclimatization to standard laboratory condition and divided into test compound and vehicle control group each having three animals. The study was conducted stepwise as follow: Starting dose 2000 mg/kg body weight: Step-I - The test compound was mixed with distilled water and administered orally at the dose level of 2000 mg/kg body weight (dose volume 10ml/kg) to three female rats. However; vehicle control group treated with the distilled water at the dose level of 10 ml/kg b.wt.  The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days.  The necropsy was performed on all animals at the termination of the study.The test compound did not elicit any clinical signs of intoxication throughout the period of observation at the tested dose level 2000 mg/kg body weight. Furthermore, no mortality was recorded at test dose level. Vehicle control group of animals were also free from any mortality and clinical signs. The necropsy finding did not reveal any gross pathological changes at the tested dose level. Furthermore, no gross pathological change was observed in vehicle control group. Step -II: After 72 hrs, the result of step-I was confirmed by administration of same dose level (2000 mg/kg. b.wt) of test compound in additional three animals of same sex (OECD-423 guidelines) under same test condition. The test compound administered at the dose level of 2000 mg/kg b.wt did not produce any mortality and clinical sign of intoxication throughout the observation period of 14 days. Body weight of each rat was also recorded on day 7th and 14th showed normal gain as compared to control group. Necropsy was conducted at the end of the study (15th day) on all the animals which did not reveal any gross pathological changes. From the results obtained from present investigation, the LD50 value was considered to be >2000 mg/kg bw. Thus, it can be concluded that the test chemical is non toxic to Wistar albino rats. CLP classification "Not classified".

 

Acute Inhalation toxicity: 

The study need not be conducted because the exposure of the test chemical via inhalation is not likely owing its low vapour pressure and/or the possibility of exposure to aerosols, particles or droplets of inhalable size.The estimated vapour pressure of test substance at 25 Deg C was 0.0000000809 Pa. Hence, this endpoint was considered for waiver.

 

Acute Dermal Toxicity:

The acute dermal toxicity study was conducted according to OECD guideline 402 for testing of chemicals on Wistar albino rats. The summary of the study was as follows -

LIMIT TEST (2000 mg/kg b.wt): Ten healthy wistar albino rats of both sex (ranging b.wt 200±20 gm) selected for study after acclimatization. Approximate 10 percent back skin of total body surface area was prepared 24 hrs prior to application of test compound. The test compound was applied dermally at the dose level of 2000 mg/kg b.wt for each animal.The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days. The body weight of each rat was observed on day 0 (pre treatment), 7th and 14th (post treatment). The necropsy was performed on all animals at termination of the study.The test compound applied at the dose level of 2000 mg/kg b.wt in Wistar albino rats did not show any mortality as well as clinical signs of toxicity throughout the observation period of 14 days.  Necropsy finding did not reveal any gross pathological changes under test condition.CONFIRMATORY TEST: After 72 hrs, a confirmatory test was conducted in same species of animals to confirm the results obtained from limit test (OECD-402 guidelines). Ten healthy Wistar albino rats of both sex (ranging b.wt 200±20 gm) selected for study after acclimatization. Approximate 10 percent back skin of total body surface area was prepared 24 hrs prior to application of test compound. The test chemical was applied dermally at the dose level of 2000 mg/kg b.wt for each animal.The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days. The body weight of each rat was observed on day 0 (pre treatment), 7th and 14th (post treatment). The necropsy was performed on all animals which was died during the study or were sacrificed at termination of the study.No mortality was recorded in Wistar albino rats after administration of test compound at the dose level of 2000 mg/kg b.wt throughout the period of observation (14 days). The test compound did not elicit any clinical signs of toxicity during the observation period. No skin reaction was observed after 24th hrs of patch removal. The body weight of each animal recorded on day 7th and 14th showed normal increase as compared to day 0 (pre treatment).From the results obtained from present investigation, the acute dermal LD50 value was considered to be >2000 mg/kg bw, when male/female Wistar albino rats were treated with the given test chemical by dermal application. Thus, it can be concluded that the test compound is non toxic to Wistar albino rats as per the criteria of CLP regulation.

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that the LD50 value is >2000 mg/kg bw, for acute oral toxicity and acute dermal toxicity and LC50 value is >5 mg/L air, for acute inhalation toxicity. Thus, comparing these values with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral, acute inhalation and acute dermal toxicity.