Registration Dossier

Toxicological information

Acute Toxicity: oral

Currently viewing:

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14 November 2018 - 13 December 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:
no

Test material

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

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: In-house bred animals
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 9 to 10 weeks
- Weight at study initiation: 177.28-205.76 (minimum and maximum mean weights of the 4 groups tested)
- Fasting period before study: Animals were fasted overnight (16 to 18 hours) prior to dosing. Water was provided ad libitum during fasting period. Feed was offered 3 to 4 hours after dosing.
- Housing: Three animals were housed in standard polypropylene cage (Size: L 430 × B 285 × H 150 mm) with stainless steel mesh top grill having facilities for holding pelleted feed and drinking water in water bottle fitted with stainless steel sipper tube. Clean sterilized paddy husk was provided as bedding material.
- Diet (e.g. ad libitum): Ad libitum. Altromin Maintenance Diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG).
- Water (e.g. ad libitum): Ad libitum. Deep bore-well water passed through Reverse osmosis unit was provided in plastic water bottles with stainless steel sipper tubes.
- Acclimation period: the animals were acclimatized for at least 5 days before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.3ºC to 22.8ºC
- Humidity (%): 46 to 66%
- Air changes (per hr): 12-15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/mL and 200 mg/mL
- Amount of vehicle (if gavage): 10 mL
- Justification for choice of vehicle: As per the in-house miscibility test, test item was miscible in corn oil. Corn oil is universally accepted and routinely used vehicle in oral toxicity studies.
- Lot/batch no. (if required): A1712001
- Purity: not reported

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg body weight

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The animals were dosed in a stepwise procedure with three female animals per step. A starting dose of 300 mg/kg body weight was selected from the fixed dose levels of 5, 50, 300 and 2000 mg/kg body weight since the LD50 for test item was unavailable.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
6 (3 female rats per step. 2 steps per dose were conducted as per OECD TG 423)
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 for clinical signs of toxicity and mortality at 30 to 40 min, 1 hr (±10 mins), 2 hrs (±10 mins), 3 hrs (±10 mins) and 4 hrs (±10 mins) post dosing on day 1 and once daily thereafter for clinical signs of toxicity and twice daily for mortality during the 14 days observation period. The body weights were recorded on day 1 (before test item administration) and on day 8 and 15 during the observation period.
- Necropsy of survivors performed: yes. At termination, gross pathological findings were recorded and reported.
- Other examinations performed:
Clinical observations included: changes in skin, fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern.

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 signs of toxicity were observed in Step-I and Step-I confirmation animals dosed with 300 mg/kg bw.
In Step-II and Step-II confirmation, animals were dosed with 2000 mg/kg bw. No clinical signs were observed on day 1; diarrhea and wet perineum was observed on day 2 and wet perineum was observed on day 3. The observed clinical signs reversed back to normal by day 4 observation.
Body weight:
No changes were observed in body weight and percent change in body weight with respect to day 1. All the animals revealed physiologically normal increase in the body weight.
Gross pathology:
No gross pathological changes were observed in any of the animals.

Any other information on results incl. tables

Table 1: Individual animal clinical signs of toxicity and mortality record

Study Steps

&

Dose

(mg/kg body weight)

Animal No.

Sex

Clinical Signs of Toxicity and Mortality on

Day 1

Clinical Signs of Toxicity and Mortality on Day

30 to 40

min

1 hr

(±10 min)

2hrs

(±10 min)

3hrs

(±10

min)

4hrs

(±10

min)

2

3

4

5

6

7

8

9

10

11

12

13

14

15

Step-I

&

300

Rd2486

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2487

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2488

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Step-I

Confirmation &

300

Rd2489

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2490

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Rd2491

F

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

N

Step-II

&

2000

Rd2492

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2493

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2494

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Step-II

Confirmation &

2000

Rd2495

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2496

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

Rd2497

F

N

N

N

N

N

36, 110

110

N

N

N

N

N

N

N

N

N

N

N

N

N: Normal; F: Female; min/mins: minutes; hr/hrs: Hour/Hours; 36: Diarrhea; 110: Wet perineum

Table 2: Individual animal body weight (g) and percent change in body weight with respect to day 1

Study Steps

&

Dose

(mg/kg body weight)

Animal No.

Sex

Body Weight (g) on Day

 

Percent Change in Body Weight with Respect to Day

1

8

15

 

1 to 8

1 to 15

Step-I

&

300

Rd2486

F

180.86

200.45

220.37

 

10.83

21.85

Rd2487

F

171.06

190.71

209.52

 

11.49

22.48

Rd2488

F

179.93

197.34

215.24

 

9.68

19.62

 

Mean

 

177.28

196.17

215.04

 

10.66

21.32

 

±SD

 

5.41

4.97

5.43

 

0.92

1.50

Step-I

Confirmation

&
300

Rd2489

F

188.74

205.41

224.83

 

8.83

19.12

Rd2490

F

191.22

210.57

229.70

 

10.12

20.12

Rd2491

F

212.24

230.23

251.49

 

8.48

18.49

 

Mean

 

197.40

215.40

235.34

 

9.14

19.25

 

±SD

 

12.91

13.10

14.20

 

0.86

0.82

Step-II

&
2000

Rd2492

F

187.11

205.49

223.81

 

9.82

19.61

Rd2493

F

202.21

220.31

241.70

 

8.95

19.53

Rd2494

F

215.18

232.84

251.93

 

8.21

17.08

 

Mean

 

201.50

219.55

239.15

 

8.99

18.74

 

±SD

 

14.05

13.69

14.23

 

0.81

1.44

Step-II

Confirmation

&
2000

Rd2495

F

209.20

230.71

258.74

 

10.28

23.68

Rd2496

F

211.92

232.42

255.27

 

9.67

20.46

Rd2497

F

196.16

215.89

239.16

 

10.06

21.92

 

Mean

 

205.76

226.34

251.06

 

10.00

22.02

 

±SD

 

8.42

9.09

10.45

 

0.31

1.61

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. 12 female Sprague-Dawley rats divided in 4 groups were administered sequentially with test item previously dissolved in corn oil by oral gavage. The first set of 3 female rats was given a single dose of 300 mg/kg bw. No mortality was observed at this dose level, so a confirmatory set of 3 female rats was treated at the same dose level. No mortality was observed at this dose level. Thus, a third set of 3 female rats was given a single dose of 2000 mg/kg bw. No mortality was observed at this dose level, so finally a confirmatory set of 3 female rats was treated at the same dose level. The body weight evolution of all animals remained normal during the study. No clinical signs of toxicity were observed in Step-I and Step-I confirmation animals dosed with 300 mg/kg bw. In Step-II and Step-II confirmation (animals dosed with 2000 mg/kg bw) no clinical signs were observed on day 1; diarrhea and wet perineum was observed on day 2 and wet perineum was observed on day 3. The observed clinical signs reversed back to normal by day 4 observation. The macroscopic examination of the animals at the end of the study did not reveal treatment related effects. Based on these results, the LD50 of the test item is deterined to be higher than 2000 mg/kg bw. As per OECD TG 423, the LD50 cut-off of the test item may be considered to be 5000 mg/kg bw.