Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 

 

The acute oral toxicity dose (LD50) was considered based on different studies conducted on rats for the test chemical. 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 can be classified as´Not classified ´.

 

Acute Inhalation Toxicity:

 

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 0.000395 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal toxicity:

 

The acute dermal toxicity dose (LD50) was considered based on different studies conducted on rats for the test chemical. The studies concluded that 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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study repport
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Principles of method if other than guideline:
The objective of this acute oral toxicity study was to assess the toxicological profile of the test item when administered to rats by a single oral gavage.
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Invivo Biosciences, Bengaluru
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 11 Weeks
- Weight at study initiation: 199.46 g to 224.12 g
- Fasting period before study: overnight fasted (16 to 18 hours)
- Housing: Rats were housed individually in standard polysulfone cages
(Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top
grill having facilities for pelleted food and drinking water in polycarbonate
bottle. Additionally, polycarbonate rat huts were placed inside the cage as an
enrichment object and were changed along with the cage at least once a week.
Bedding: steam sterilized corn cob was used and changed at least once a
week along with the cage.
- Diet (e.g. ad libitum): Rat & Mice pellet feed, manufactured by Krishna Valley Agro Tech LLP
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed
to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by
Eureka Forbes Ltd, Mumbai
- Acclimation period: After physical examination for good health and
suitability for experiment, the animals were
acclimatized six to twenty two days prior to
treatment. Animals were observed once daily during
acclimatization period. Females were nulliparous
and non-pregnant.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24°C
- Humidity (%): 64 to 66%,
- Air changes (per hr): 13.2-13.8 air changes/hour.
- Photoperiod (hrs dark / hrs light): 12 hours light and
12 hours dark cycle.

IN-LIFE DATES: From: To:31.08.2018 to 06.10.2018
Route of administration:
oral: gavage
Vehicle:
other: 0.5% w/v Sodium carboxy methyl cellulose in Milli-Q water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 550, 730, 970, 1290,1750 and 2300 mg/kg
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle: Based on solubility test, the test item forms a suspension in 0.5% sodium
carboxy methyl cellulose (NaCMC) in Milli-Q water and used to prepare the
dose formulations.
- Lot/batch no. (if required): SLBR1692V

MAXIMUM DOSE VOLUME APPLIED: 2300 mg/kg

DOSAGE PREPARATION (if unusual):A required quantity (g) of test item was weighed in an aluminum foil and transferred into clean mortar and gently triturated using pestle to obtain fine powder. Small volume of vehicle was added gradually to the mortar with continuous trituration until a uniform suspension was obtained. The mixture was quantitatively transferred to a measuring cylinder. Further, a small volume of vehicle was added to the mortar and rinsed with the vehicle, all the rinsing was quantitatively transferred into the measuring cylinder. The final volume was made up to the mark with vehicle to get the desired test item concentration (mg/mL). The prepared dose formulation was transferred to the dedicated labeled beaker. Preparation were made prior to dosing.Homogeneity of the test item in the vehicle was maintained during treatment by constant stirring using a magnetic stirrer.
Doses:
G1A/ 1-550 mg/kg
G2/2- 730 mg/kg
G3/3- 970 mg/kg
G4/4- 1290 mg/kg
G5/5- 1750 mg/kg
G6/6- 2300 mg/kg
No. of animals per sex per dose:
1 female per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At each step, the animals were observed five times on test day 1 (day of
administration) i.e. at 30 minutes and four times at hourly intervals and once
daily during days 2 to 15 post administration.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:
Clinical signs:Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to the observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma and all observed clinical signs were recorded.

Body weights
The body weights were recorded on test day 1 (pre-administration), day 8
(7 days post administration) and day 15 (14 days post administration).

Necropsy
The rats surviving to the end of the observation period were euthanized by
using isoflurane anaesthesia and subjected to detailed necropsy. Gross
pathological findings were recorded and reported. Microscopic examination
was not carried out as no gross pathological changes were observed.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 300 mg/kg bw
Based on:
test mat.
Remarks on result:
other:
Remarks:
No mortality observed
Mortality:
No mortality observed
Clinical signs:
G1/Step-1 to G6/Step-6: There were no clinical sings and pre-terminal
deaths.
Body weight:
G1/Step-1 to G6/Step-6: The body weights of all the rats increased
throughout the observation period.
Gross pathology:
There were no gross pathological changes at necropsy
Other findings:
Not specified

Table 3:Body weight, body weight change and pre-terminal deaths

 

Group/Step and Dose (mg/kg

body weight)

Rat No.

Sex

Body weight (g)

Day of Death(Time of

Death)

No. dead/

No.

tested

Preterminal deaths (%)

 

Initial

(Day 1)

8th day

Weight

change

(day 8 –

Initial)

15th day

 

Weight

change

(day 15

– Initial)

At Death

G1/Step 1/550

Rw979

F

221.37

225.61

4.24

230.26

8.89

NA

NA

NIL

-

G1A/Step 1/550

Rw980

F

224.12

226.76

2.64

231.79

7.67

NA

NA

NIL

-

G2/Step 2/730

Rw981

F

210.84

213.26

2.42

221.12

10.28

NA

NA

NIL

-

G3/Step 3/970

Rw982

F

209.16

214.03

4.87

220.24

11.08

NA

NA

NIL

-

G4/Step 4/1290

Rw983

F

201.90

214.17

12.27

219.38

17.48

NA

NA

NIL

-

G5/Step 5/1750

Rw984

F

206.64

210.32

3.68

214.96

8.32

NA

NA

NIL

-

G6/Step 6/2300

Rw985

F

199.46

203.61

4.15

208.52

9.06

NA

NA

NIL

-

F: Female NA: Not Applicable mg: milligram kg: kilogram g: gram

Note: G1/Step 1 was not considered for results and assessment of toxicity as the food was provided before dosing

Interpretation of results:
other: Not classified
Conclusions:
Based on the results of the present study, the LD50 for test chemical is greater than 2000 mg/kg body weight
Executive summary:

The acute oral toxicity study was conducted to assess the toxicological profile of the test item as per OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure) in Wistar rats.

The dose formulation was prepared by using Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) female rats at each step. There were no clinical signs of toxicity and pre-terminal deaths.

The stopping criteria met as 3 consecutive animals survive at the upper bound. Hence, the further dosing was stopped as per AOT 425 Stat program result (version: 1.0). There were no clinical signs of toxicity and pre-terminal deaths.

The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and on days 8 and 15. Necropsy was performed for all the rats at termination. All survived rats gained weight during experimental period. There were no gross pathological changes at necropsy.

Based on the results of the present study, the LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute oral toxicity. CLP criteria "Not Classified".

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 300 mg/kg bw
Quality of whole database:
Data is Klimisch 1 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:
other:
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:
1 (reliable without restriction)
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:
The objective of this acute dermal toxicity study was to assess the toxicological profile of the test item on application as a single semi-occlusive dermal application to rats.
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Invivo Biosciences Bengaluru
- Females (if applicable) nulliparous and non-pregnant:yes
- Age at study initiation: 8 to 10 weeks
- Weight at study initiation: 200.06 to 238.66 g
- Fasting period before study:
- Housing: Animals were housed individually in standard polysulfone cages
(Size: approximately L 425 x B 266 x H 185 mm), with stainless steel top
grill having facilities for pelleted food and drinking water in polycarbonate
bottle. Additionally, polycarbonate rat huts were placed inside the cage as
enrichment objects and were changed along with the cage at least once a
week.
Bedding: Steam sterilized corn cob was used and changed at least once a
week along with the cage.
- Diet (e.g. ad libitum): Hypro Rat & Mice pellet feed, manufactured by Krishna Valley Agro Tech, was provided to animals.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by
Eureka Forbes Ltd, Mumbai , India, was provided to animals in polycarbonate bottles with stainless steel sipper tubes
- Acclimation period: After physical examination for good health and suitability for experiment, the rats were acclimatized for seven days for G1DRF, eleven days for G2DRF, fifteen days for G3DRF, eighteen days for G3 Main group before treatment under standard laboratory conditions. Animals were observed once daily during acclimatization period. Females were nulliparous and non-pregnant

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 24°C,
- Humidity (%): 64 to 66 %
- Air changes (per hr): 13.2-13.8 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours
light and 12 hours dark cycle

IN-LIFE DATES: From: To:31.08.2018-02.10.2018
Type of coverage:
semiocclusive
Vehicle:
other: Milli-Q water
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsolateral thoracic surface of the skin
- % coverage: 10 %
- Type of wrap if used: adhesive tape and cotton gauge

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washed with deionized water and wiped
dry using clean towel
- Time after start of exposure:24 hrs

TEST MATERIAL
- Amount(s) applied :200, 1000 and 2000 mg/kg bw
- Concentration (if solution):
- Constant volume or concentration used: no
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit):10 mg/l
Duration of exposure:
24 hours
Doses:
G1: 200 mg/kg bw
G2: 1000 mg/kg bw
G3: 2000 mg/kg bw
No. of animals per sex per dose:
1 female per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:The animals were observed for clinical signs and pre-terminal deaths (mortality) once during first 30 minutes after application, and at hourly intervals for 6 hours after application on the day of treatment (day 1) and once daily during Days 2 to 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:treatment site was observed 24, 48 and 72 hours after removal of test chemical using the Draize criteria (Refer Annexure 4 of this report). All rats were observed for changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Statistics:
Not specified
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other:
Remarks:
No mortality observed
Mortality:
No mortality observed
Clinical signs:
There were no clinical signs and pre-terminal deaths (mortality) observed during the study. There were no skin reactions at the site of application at 24, 48 and 72 hours after test patch removal (as per draize method).
Body weight:
All rats gained body weight throughout the observation period
Gross pathology:
No abnormality was detected at necropsy

Table 3: Individual body weight, body weight changes and pre-terminal deaths

Group and Dose

(mg/kg body weight)

Rat No.

Sex

Body weight (g)

Pre-terminal deaths

Initial deaths

(Day 1 - at

treatment)

8thday

Weight change

(day 8 – Initial)

15thday

Weight change

(day 15 – Initial)

G1and200DRF

Rw991

F

238.66

243.39

4.73

246.84

8.18

0

G2and1000DRF

Rw992

F

209.41

213.09

3.68

222.64

13.23

0

G3and2000DRF

Rw993

F

207.43

210.56

3.13

215.41

7.98

0

G3and2000Mainstudy

Rw994

F

201.54

204.86

3.32

209.47

7.93

0

Rw995

F

200.06

202.45

2.39

205.79

5.73

0

DRF: Dose Range Finding    F: Female

Table 4: Individual test item application, clinical signs, skin reaction and necropsy findings

Dose range finding study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G1 and 200

7 September

2018

and

11.07 AM

Rw991

F

238.66

47.73

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G1 and 200

Rw991

F

N

N

N

N

N

N

N

N

N

N

NAD

 

 

Table 5:Dose range finding study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G2 and 1000

11 September

2018

and

11.54 AM

Rw992

F

209.41

209

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G2 and 1000

Rw992

F

N

N

N

N

N

N

N

N

N

N

NAD

 

 Table 6:Dose range finding study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G3 and 2000

14 September

2018

and

11.18 AM

Rw993

F

207.43

415

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G3 and 2000

Rw993

F

N

N

N

N

N

N

N

N

N

N

NAD

 

 

Table 7:Main study

 

Group &

Dose

(mg/kg

body weight)

Date and time

of application

Rat

Number

Sex

Initial

Bwt

(g)

Quantity

(mg)

applied

Observation and skin reaction

Days

1

2

3

4

5

30 min

1h

2h

3h

4h

5h

6h

*

Er@

Ed@

*

Er@

Ed@

*

Er@

Ed@

G3 and 2000

18 September

2018

and

10.1 AM and 10.4 AM

Rw994

F

201.54

403

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Rw995

F

200.06

400

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

 

 

Group & Dose

(mg/kg

body weight)

Animal

Number

Sex

Observation

Necropsy findings

Days

6

7

8

9

10

11

12

13

14

15

G3 and 2000

Rw994

F

N

N

N

N

N

N

N

N

N

N

NAD

 

Rw995

F

N

N

N

N

N

N

N

N

N

N

NAD

F: Female

N: Normal

h: hour

min: minutes

NAD: No abnormality detected

Er: Erythema

Ed: Edema Score

0: No Erythema / Edema

*: Clinical signs; @: Skin scoring as per Draize method (approximately 24, 48 and 72 hours) after test patch removal

Interpretation of results:
other: Not classified
Conclusions:
Based on the present study results, the acute dermal LD50 of test chemical is more than 2000 mg/kg body weight in female Wistar rats.
Executive summary:

The acute dermal toxicity study was conducted to assess the toxicological profile of the test item as per OECD Guideline 402 (Acute Dermal Toxicity) in Wistar rats.

The test item at the dose of 200, 1000 and 2000 mg/kg body weight was transferred on to the cotton gauze and applied directly (semi-occlusive) to the clipped skin of the animal to cover about 10% of the body surface of the rat It was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was for 24 hours. There were no clinical signs of toxicity and pre-terminal deaths.

All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the treatment site was observed for skin reactions at 24, 48 and 72 hours after removal of test item using the Draize criteria. There were no clinical signs of toxicity and mortality observed. There was no skin reaction observed at test item applied area. Body weight was measured on days 1, 8 and 15 and all rats gained weight during experimental period. At the end of observation period, all surviving animals were euthanized and subjected to necropsy. There were no abnormalities detected at the necropsy.

Based on the results of the present study, the LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute dermal toxicity. CLP criteria "Not Classified".

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from study report

Additional information

Acute oral toxicity:

 

In different studies, the given test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats and mice for test chemical. The studies are summarized as below –

 

1.The acute oral toxicity study was conducted to assess the toxicological profile of the test item as per OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure) in Wistar rats.

The dose formulation was prepared by using Milli-Q water and administered as a single oral gavage to overnight fasted (16 to 18 hours) female rats at each step. There were no clinical signs of toxicity and pre-terminal deaths.

The stopping criteria met as 3 consecutive animals survive at the upper bound. Hence, the further dosing was stopped as per AOT 425 Stat program result (version: 1.0). There were no clinical signs of toxicity and pre-terminal deaths.

The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and on days 8 and 15. Necropsy was performed for all the rats at termination. All survived rats gained weight during experimental period. There were no gross pathological changes at necropsy.

Based on the results of the present study, the LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute oral toxicity. CLP criteria "Not Classified".

In another study Acute oral toxicity study of test chemical was conducted on mice at the dose concentration of 599 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 14 days.Thus, LD50 value was considered to be 599 mg/kg bw, when mice were treated with test chemical via oral route.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw according to experimental report. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified ´Not classified ´

 

Acute Inhalation Toxicity:

 

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 0.000395 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal Toxicity:

 

  The acute dermal toxicity study was conducted to assess the toxicological profile of the test item as per OECD Guideline 402 (Acute Dermal Toxicity) in Wistar rats.

The test item at the dose of 200, 1000 and 2000 mg/kg body weight was transferred on to the cotton gauze and applied directly (semi-occlusive) to the clipped skin of the animal to cover about 10% of the body surface of the rat It was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was for 24 hours. There were no clinical signs of toxicity and pre-terminal deaths.

All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the treatment site was observed for skin reactions at 24, 48 and 72 hours after removal of test item using the Draize criteria. There were no clinical signs of toxicity and mortality observed. There was no skin reaction observed at test item applied area. Body weight was measured on days 1, 8 and 15 and all rats gained weight during experimental period. At the end of observation period, all surviving animals were euthanized and subjected to necropsy. There were no abnormalities detected at the necropsy.

Based on the results of the present study, the LD50 value of the given test chemical is >2000 mg/kg bw. Thus, the test item is not classified for acute dermal toxicity. CLP criteria "Not Classified".

 

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

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw for acute oral and dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified as not classified for acute oral and dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.