Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report Date:
2017

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Geniron Biolabs Pvt. Ltd., No.93, Solur, Anekal - Thally Road, Anekal, Bengaluru – 562106, India

- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 9 to 10 weeks
- Weight at study initiation: Males : 270.8 to 285.3 g ; Females: 220.3 to 229.4 g
- Fasting period before study: No
- Housing: Animals were housed individually in standard polysulfone cages
- Diet (e.g. ad libitum): Hypro rat & mice pellet feed, manufactured by Pranav Agro Industries Ltd., Pune 411 030, Maharashtra, India
- 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 400 001, India
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 25°C
- Humidity (%): 64 to 67 %
- Air changes (per hr): 14.1 to 14.3 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle

IN-LIFE DATES: From: 28-Feb-2017 To: 20-Mar-2017

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
other: Milli-Q water
Details on dermal exposure:
TEST SITE
- Area of exposure: Males: 9 x 6 cm; Females: 8 x 5 cm
- % coverage: 10% of body surface of the rat
- Type of wrap if used: Cotton gauze was secured in position by adhesive tape wound around the torso

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Water
- Time after start of exposure: Approximately 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Refer Appendix 1
- For solids, paste formed: Yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.4 mL
Duration of exposure:
24 hours
Doses:
2000 mg/kg body weight
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 (or other?)
- 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. Body weight on days 1, 8 and 15.
- Necropsy of survivors performed: Yes
- Other examinations performed: clinical signs, body weight,gross pathology

Results and discussion

Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
no indication of skin irritation up to the relevant limit dose level
Mortality:
No mortality
Clinical signs:
There were no clinical signs observed during the study. However, blue colour stain was observed on test item application area in all rats on day 2 and 3 post dose observation
Body weight:
All rats gained body weight throughout the observation period
Gross pathology:
No abnormality was detected at necropsy

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 (%)

Initial

(at treatment)

8th  

day

Weight change

(day 8 – Initial)

15th

day

Weight change

(day 15 – Initial)

 

G1 and

2000

Limit test

Rm5881

M

278.9

294.8

15.9

316.8

37.9

 

0/10

 

0

Rm5882

M

285.3

295.1

9.8

315.7

30.4

Rm5883

M

283.5

299.6

16.1

321.2

37.7

Rm5884

M

284.4

295.7

11.3

314.4

30.0

Rm5885

M

270.8

282.7

11.9

310.7

39.9

Rm5886

F

220.3

227.9

7.6

236.2

15.9

Rm5887

F

220.5

228.5

8.0

233.9

13.4

Rm5888

F

229.4

240.9

11.5

249.7

20.3

Rm5889

F

221.4

230.1

8.7

237.2

15.8

Rm5890

F

222.9

229.6

6.7

238.6

15.7

F: Female            M: Male   

APPENDIX 1.      Individual test item application, clinical signs and necropsy findings

Group and Dose

(mg/kg body weight)

Date &

Time of

Application

Rat

No.

Sex

Body weight initial

(g)

Quantity

 applied

(mg)

 

Day of Observations

Day 1

2

3

1

hour

2

hours

3

hours

4

hours

 

 

AM

PM

AM

PM

G1

and

2000

Limit test

06 March 2017

&

10.42 AM

to

10.49 AM

Rm5881

M

278.9

558

N

N

N

N

N*

N*

N*

N*

Rm5882

M

285.3

571

N

N

N

N

N*

N*

N*

N*

Rm5883

M

283.5

567

N

N

N

N

N*

N*

N*

N*

Rm5884

M

284.4

569

N

N

N

N

N*

N*

N*

N*

Rm5885

M

270.8

542

N

N

N

N

N*

N*

N*

N*

Rm5886

F

220.3

441

N

N

N

N

N*

N*

N*

N*

Rm5887

F

220.5

441

N

N

N

N

N*

N*

N*

N*

Rm5888

F

229.4

459

N

N

N

N

N*

N*

N*

N*

Rm5889

F

221.4

443

N

N

N

N

N*

N*

N*

N*

Rm5890

F

222.9

446

N

N

N

N

N*

N*

N*

N*

F: Female        M: Male         N: Normal          AM: Ante Meridian                 PM: Post Meridian    

*:Blue colour stain on the test item application area.

APPENDIX 1 contd. Individual test item application, clinical signs and necropsy findings

 

Group and Dose

(mg/kg body weight)

Rat

No.

Sex

Day of Observations

Necropsy

Findings

4

5

6

7

8

9

10

11

12

13

14

15

G1

and

2000

Limit test

Rm5881

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5882

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5883

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5884

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5885

M

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5886

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5887

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5888

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5889

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm5890

F

N

N

N

N

N

N

N

N

N

N

N

N

NAD

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



Applicant's summary and conclusion

Interpretation of results:
other: not classified
Conclusions:
Based on the present study results, single dermal application of the test item was associated with no mortality and neither signs of toxicity nor signs of irritation.
The acute dermal LD50 of the test item is more than 2000 mg/kg body weight in male and female Wistar rats.
Executive summary:

The acute dermal toxicity of the test item was tested in 5 male and 5 female Wistar rats at the limit dose of 2000 mg/kg body weight.

Based on the individual body weight, the finely ground test item at the dose of 2000 mg/kg body weight was weighed on an aluminium foil and made into a paste by adding sufficient volume of the Milli-Q water (approximately 0.4 mL) and completely transferred on to the cotton gauze (size: Males: 9 x 6 cm; Females: 8 x 5 cm of 6 ply) and applied                  (semi-occlusive) directly to the clipped skin of the rat to cover about 10% of body surface of the rat. Then the cotton gauze was secured in position by adhesive tape wound around the torso. The test item contact period with the skin was 24 hours.

After the 24 hour contact period, the adhesive tape and cotton gauze were removed and the application area was washed with water and wiped dry using a clean towel. 

All the rats were observed for clinical signs of toxicity and mortality for
14 days post application. There were no clinical signs of toxicity and mortality. However, blue colour stain was observed at the test item application area in all rats on day 2 and 3 post dose observation. 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 necropsy
.

Based on the present study results, single dermal application of the test item was associated with no mortality and neither signs of toxicity nor signs of irritation.

The acute dermal LD50of the test item is more than 2000 mg/kg body weight in male and female Wistar rats.