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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 February 2018 - 07 March 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:In-Vivo Biosciences, Kodigehalli Village, Magadi Road, Bangalore, Code-29, Karnataka State
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 11 weeks
- Weight at study initiation: 187.8 to 201.6 g
- Fasting period before study: overnight prior to dosing until 3-4 hours post-dosing
- 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 once a week. Bedding: steam sterilized corn cob was used and changed once a week along with the cage.
- Diet (e.g. ad libitum): Ad libitum. Hypro Rat & Mice pellet feed, manufactured by Krishna Valley Agro Tech LLP, MIDC Kupwad block, Sangli, Maharashtra, was provided to animals.
- Water (e.g. ad libitum): 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 400 001, India, was provided to animals in polycarbonate bottles with stainless steel sipper tubes.
- Acclimation period: the animals were acclimatized for six days (G1-first treatment step) and for eight days (G1-second treatment step) before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24ºC
- Humidity (%): 64 to 67%
- Air changes (per hr): 13.5 to 13.6 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 2.12 mL/kg (Calculated based on the density of the test item: 0.945 g/mL (as per TIDS and COA))

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Available information suggests that mortality is unlikely at the highest starting dose level (2000 mg/kg body weight). Hence, the study is initiated with the starting dose of 2000 mg/kg body weight.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6
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. 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 of survivors performed: yes. At termination, gross pathological findings were recorded and reported.
- Other examinations performed:
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.

Results and discussion

Effect levelsopen allclose all
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Key result
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred during the study.
Clinical signs:
No clinical sign was observed in rats treated with 2000 mg/kg body weight.
Body weight:
Normal gain in body weight was observed in all the rats treated with 2000 mg/kg body weight.
Gross pathology:
There were no gross pathological changes at necropsy.

Any other information on results incl. tables

Table 1. Body weight, body weight change and pre-terminal deaths

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Body weight (g)

Day of Death (Time of Death)

No. dead/

 No. tested

Pre-terminal deaths

(%)

Initial

(Day 1)

8thday

Weight change

(day 8 – Initial)

15thday

Weight change          

(day 15 – Initial)

At Death

G1

(FTS)

2000

(2.12 mL/kg *)

 

Rm8055

F

196.2

203.9

7.7

215.0

18.8

NA

NA

 

0/3

 

0

Rm8056

F

188.6

198.7

10.1

210.3

21.7

NA

NA

Rm8057

F

187.8

198.1

10.3

210.8

23.0

NA

NA

G1

(STS)

2000

(2.12 mL/kg *)

 

Rm8058

F

192.8

203.6

10.8

214.2

21.4

NA

NA

 

0/3

 

0

Rm8059

F

188.5

199.7

11.2

211.6

23.1

NA

NA

Rm8060

F

201.6

210.9

9.3

222.5

20.9

NA

NA

FTS: First Treatment Step; STS: Second Treatment Step

*: Calculated based on the density of the test item: 0.945 g/mL (as per TIDS and COA).

Table 2. Individual clinical signs, dose administration and necropsy findings

Group and

Dose

(mg/kg

body weight)

Date and

time of administration

Rat

No.

Sex

Body

weight

(Day 1)

Total volume administered

(mL)

Day of observation

Day 1

30 min

1hour

2 hours

3 hours

4 hours

G1

(FTS)

2000

(2.12 mL/kg*)

19 February 2018 and

09:55 AM

to

09.57 AM

Rm8055

F

196.2

0.42

N

N

N

N

N

Rm8056

F

188.6

0.40

N

N

N

N

N

Rm8057

F

187.8

0.40

N

N

N

N

N

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Day of observation

Necropsy

Findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G1

(FTS)

2000

(2.12 mL/kg*)

Rm8055

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm8056

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm8057

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Group and

Dose

(mg/kg

body weight)

Date and

time of administration

Rat

No.

Sex

Body

weight

(Day 1)

Total volume administered

(mL)

Day of observation

Day 1

30 min

1hour

2 hours

3 hours

4 hours

G1

(STS)

2000

(2.12 mL/kg*)

21 February 2018 and

09.53 AM

to

09.55 AM

Rm8058

F

192.8

0.41

N

N

N

N

N

Rm8059

F

188.5

0.40

N

N

N

N

N

Rm8060

F

201.6

0.43

N

N

N

N

N

Group and

Dose

(mg/kg

body weight)

Rat

No.

Sex

Day of observation

Necropsy

Findings

2

3

4

5

6

7

8

9

10

11

12

13

14

15

G1

(STS)

2000

(2.12 mL/kg*)

Rm8058

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm8059

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm8060

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

FTS: First Treatment Step; STS: Second Treatment Step; NAD: No Abnormality Detected

*: Calculated based on the density of the test item: 0.945 g/mL (as per TIDS and COA).

Applicant's summary and conclusion

Interpretation of results:
other: Not classified (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. The LD50 cut-off may be considered to be 5000 mg/kg body weight by oral route in the rat.
Executive summary:

The acute oral toxicity of the test item has been tested in accordance with OECD Test Guideline 423, following GLP. 6 female Wistar rats divided in 2 groups were administered sequentially with test item by oral gavage. The first set of 3 female rats was given a single dose of 2000 mg/kg body weight. No mortality was observed at this dose level, so the second set of 3 female rats was treated at the same dose level. No mortality was observed at this dose level either. The body weight evolution of the animals remained normal during the study. No clinical signs were observed. The macroscopic examination of the animals at the end of the study did not reveal treatment related effects. The LD50 of the test item is higher than 2000 mg/kg body weight by oral route in the rat. As per OECD Test Guideline 423, the LD50 cut-off of the test item may be considered to be 5000 mg/kg body weight by oral route in the rat.