Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
27/06/2016-19/07/2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
The test material corresponded to the approximately 45% of the registration substance. The given dose refers to the amount of the registration substance.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Sainath Agencies, Bapujinagar, Musheerabad, Hyderabad 500 020

- Age at study initiation: 8 - 9 Weeks

- Weight at study initiation: Females: 211.6 to 229.2 g & Males: 235.8 to 266.8 g


- 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 an enrichment object and were changed along with the cage at least once a week. Bedding: steam sterilized corn cob was used and changed once a week along with the cage.

- Diet (ad libitum):Hypro Rat & Mice Pellet Feed, manufactured by Pranav Agro Industries Ltd., Pune 411 030, Maharashtra, India, was provided to animals.

- Water (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: After physical examination, the animals were acclimatized for five days for females and seven days for males before treatment. Animals were observed once daily during acclimatization period. Females were nulliparous and non-pregnant.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 23°C
- Humidity (%):65 to 67%
- Air changes (per hr):12.9 to 13.1 air changes/hour
- Photoperiod (hrs dark / hrs light):12 hours light and 12 hours dark cycle

IN-LIFE DATES: From: 27 May 2016 To: 17 June 2016

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Remarks:
The applied dose level refers to the active ingredient, the registration substance.
Details on dermal exposure:
TEST SITE
- Area of exposure: approximately 10 x 8 cm
- % coverage: 10%
- Type of wrap if used: Adhesive tape

REMOVAL OF TEST SUBSTANCE
- Washing: with water
- Time after start of exposure: after 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Refer Table 1.
Based on the individual body weight, the undiluted test item at the dose of 2000 mg/kg body weight and dose volume was 4.52 mL/kg body weight
[1.90 mL/kg (dose volume as per density) x 2.38 (correction factor)] was calculated based on the density of the test item i.e., 1.05 g/cm3 (as per TIDS provided by the sponsor). For example, for rat Rm4741, the body weight was 222.4 g, the dose volume administered was 0.87 mL [i.e., body weight of rat 222.4 g x dose volume 4.52 mL/kg / 1000 = 222.4 x 4.52 / 1000 = 1.005 mL rounded off to 1.01 mL]

- Concentration (if solution): Undiluted test item
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 per sex
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) four times (at hourly intervals after application) during day 1 and twice daily on day 2 and 3 and once daily during days 4 to 15. Individual body weights of animals were recorded on test days 1 (Pre-application), 8 (7 days post application), and 15 (14 days post application).
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, gross pathology.

Results and discussion

Preliminary study:
Body weights, body weight changes and pre-terminal deaths are presented in Tables 1.
Individual clinical signs of toxicity and necropsy findings are presented in Table 2.
Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No Mortality
Clinical signs:
Refer Table 2.
There were no clinical signs observed during the study. However, in females, the skin reactions of erythema, edema, scale formation, peeling/desquamation were observed during days 3 to 10 post dose application. The rat numbers Rm4741 and Rm4745 were normal from day 7 onwards, Rm4742 was normal from day 8 onwards, Rm4743 was normal from day 10 onwards and rat Rm4744 were normal from day 11 onwards.
Body weight:
All rats gained body weight throughout the observation period when compared to their initial weight.
Gross pathology:
No abnormalities detected.

Any other information on results incl. tables

Table 1. Individual Body Weight, Body Weight Changes and Pre-Terminal Deaths

Group and

Dose

(mg/kg

body weight)

Rat

No.

S

e

x

Body weight (g)

No. dead /

No. tested

Pre- terminal deaths (%)

Day 1

Initial

(at treatment)

8th  

day

Weight change

(day 8 – Initial)

15th

 day

Weight change

(day 15 – Initial)

 

 

 

 

 

 

 

G1

2000

(4.52 mL/kg)*

 

 

Rm4741

F

222.4

231.6

9.2

239.8

17.4

 

 

 

 

 

 

 

 

0/10

 

 

 

 

 

 

 

 

0

Rm4742

F

229.2

242.3

13.1

247.6

18.4

Rm4743

F

224.6

230.3

5.7

236.9

12.3

Rm4744

F

211.6

218.9

7.3

222.7

11.1

Rm4745

F

218.2

229.5

11.3

238.5

20.3

Rm4746

M

235.8

247.2

11.4

259.4

23.6

Rm4747

M

240.3

251.9

11.6

263.1

22.8

Rm4748

M

266.8

276.9

10.1

287.5

20.7

Rm4749

M

262.2

277.1

14.9

286.5

24.3

Rm4750

M

255.8

272.7

16.9

281.8

26.0

F : Female         M: Male   

 

*: Calculated based on the density of the test item: 1.05 g/cm3(as per TIDS provided by the sponsor), i.e., 1.90 mL/kg (dose volume as per density) x 2.38 (correction factor).

Table 2.        Individual Clinical / Toxic Signs and Necropsy Findings

Rat

No.

Sex

Body weight initial

(g)

Volume

 applied

(mL)

 

Day of Observations

Day 1

2

3

4

5

6

7

1

hour

2

hours

3

hours

4

hours

 

 

 

 

 

 

 

AM

PM

AM

PM

Rm4741

F

222.4

1.01

N

N

N

N

N

N

124(4)

124(4)

124(1)

124(2)

124(2)

N

Rm4742

F

229.2

1.04

N

N

N

N

N

N

124(4)

124(4)

124(1)

124(1)

124(2)

124(2)

Rm4743

F

224.6

1.02

N

N

N

N

N

N

124(4)

124(5)

124(4)

124(5)

124(4)

124(1)

124(1)

124(2)

Rm4744

F

211.6

0.96

N

N

N

N

N

N

124(4)

124(5)

124(4)

124(5)

124(4)

124(1)

124(1)

124(2)

Rm4745

F

218.2

0.99

N

N

N

N

N

N

124(4)

124(4)

124(1)

124(2)

124(2)

N

Rm4746

M

235.8

1.07

N

N

N

N

N

N

N

N

N

N

N

N

Rm4747

M

240.3

1.09

N

N

N

N

N

N

N

N

N

N

N

N

Rm4748

M

266.8

1.21

N

N

N

N

N

N

N

N

N

N

N

N

Rm4749

M

262.2

1.19

N

N

N

N

N

N

N

N

N

N

N

N

Rm4750

M

255.8

1.16

N

N

N

N

N

N

N

N

N

N

N

N

F: Female            M: Male               NAD: No Abnormality Detected      N:                      AM: Ante Meridian      PM: Post    

124(1): Scale formation           124(2): peeling / desquamation              124(4): Erythema           124(5): Edema

 

*: Calculated based on the density of the test item: 1.05 g/cm3(as per TIDS provided by the sponsor), i.e., 1.90 mL/kg (dose volume as per density) x 2.38 (correction factor).

Table 2 contd. Individual Clinical / Toxic Signs and Necropsy Findings

 Group: G1                                                                Dose: 2000 mg/kg body weight (4.52 mL/kg *)

Rat

No.

Sex

Body weight initial

(g)

Volume

 applied

(mL)

 

Days of observation

Necropsy

Findings

 

8

9

10

11

12

13

14

15

 

 

 

 

 

 

 

 

 

Rm4741

F

222.4

1.01

N

N

N

N

N

N

N

N

NAD

 

Rm4742

F

229.2

1.04

N

N

N

N

N

N

N

N

NAD

 

Rm4743

F

224.6

1.02

124(2)

124(2)

N

N

N

N

N

N

NAD

 

Rm4744

F

211.6

0.96

124(2)

124(2)

124(2)

N

N

N

N

N

NAD

 

Rm4745

F

218.2

0.99

N

N

N

N

N

N

N

N

NAD

 

Rm4746

M

235.8

1.07

N

N

N

N

N

N

N

N

NAD

 

Rm4747

M

240.3

1.09

N

N

N

N

N

N

N

N

NAD

 

Rm4748

M

266.8

1.21

N

N

N

N

N

N

N

N

NAD

 

Rm4749

M

262.2

1.19

N

N

N

N

N

N

N

N

NAD

 

Rm4750

M

255.8

1.16

N

N

N

N

N

N

N

N

NAD

 

 F: Female            M: Male               NAD: No Abnormality Detected      N:                      AM: Ante    

  PM: Post Meridian        124(1): Scale formation           124(2): peeling/desquamation          124(4): Erythema           124(5): Edema

 

*: Calculated based on the density of the test item: 1.05 g/cm3(as per TIDS provided by the sponsor), i.e., 1.90 mL/kg (dose volume as per density) x

    2.38 (correction factor).

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results of the present study, the LD50 of the registration substance is determined to be higher than 2000 mg/kg bw.
Executive summary:

The acute dermal toxicity of the registration substance was investigated according to the OECD Gudieline 402. Five female and five male rats were treated once with the registration substance at dose of 2000 mg/kg bw. No effect was observed. The LD50 of the registration substance was determined to be higher than 2000 mg/kg bw. No classification is warranted.