Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
09/07/2019-03/09/2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
OECD 402

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report Date:
2019

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
09th October 2017
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
impurity
Test material form:
not specified
Details on test material:
Not specified

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
3 female / group (1 for dose range finding study and 2 for main study)

Source : Invivo Biosciences, Shed No. 23, Khatha No.3169, Assessment No. 154, Kodigehalli Village, Magadi Road,
Bengaluru-560091.
Age at treatment: 09 to 10 weeks
Body weight range at treatment: Females: 209.3 to 211.5 g
Identification : By rat accession number. Identification of individual rats is by cage card and crystal violet and turmeric solution body
markings. The temporary body marking during acclimatization period was done with crystal violet.
The rat accession numbers were allotted during the course of the study and was included in raw data and reported.
Acclimatization: After physical examination for good health and suitability for experiment, the rats were acclimatized for six and ten days
before treatment for dose range finding and main study respectively under standard laboratory conditions.
Animals were observed once daily during acclimatization period. Females were nulliparous and non-pregnant.
Only healthy and intact skin rats free of clinical signs were used for this study.

Animal care practices were conformed to the requirements of the CPCSEA.
This protocol was approved by Institutional Animal Ethics Committee (IAEC) of Eurofins Advinus Limited (Proposal No. 002/Aug-2018, dated:
28 August 2018).

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
0.4 mL of the milli-Q water
Details on dermal exposure:
Approximately 24 hours before treatment, the hair on the dorsolateral thoracic surface of the skin was clipped (approximately 10 x 8 cm) with an electric clipper (Aesculap - Germany). Care was taken to avoid abrading the skin, which could alter its permeability.
Based on the individual body weight, the test item at the dose of 2000 mg/kg body weight was weighed on an aluminium foil and made into a paste by adding 0.4 mL of the milli-Q water and completely transferred on to the cotton gauze (size: Females: 8 x 5 cm of 6 ply) and applied directly (semi-occlusive) to the clipped skin of the rat to cover about 10% of body surface of the rat. It was secured in position by adhesive tape wound around the torso.
Duration of exposure:
The test item contact period with the skin was for 24 hours.
After the 24 hours contact period, the dressing was removed and the applied area was washed with deionised water and wiped dry using clean towel.
Doses:
As per the MSDS provided by sponsor, the acute dermal toxicity for Pyrogallol\ Pyrogallic acid in rats is >2000 mg/kg. Hence, an initial limit dose of 2000 mg/kg body weight was tested with 1 female rat (dose range finding study). As there was no mortality at dose range finding study. The main study was conducted with 2 further animals to confirm the classification outcome. There was no test item-related mortality. The subsequent dosing was done at least 48 hours after the previous dosing.
No. of animals per sex per dose:
3 female / group (1 for dose range finding study and 2 for main study) all trated whit 2000 mg/kg of test item.
Control animals:
not required

Results and discussion

Preliminary study:
An initial limit dose of 2000 mg/kg body weight was tested with 1 female rat (dose range finding study). As there was no mortality at dose range finding study. The main study was conducted with 2 further animals to confirm the classification outcome. There was no test item-related mortality.
Effect levels
Key result
Sex:
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:
There was no mortality observed at 2000 mg/kg body weight.
Clinical signs:
There were no clinical signs in any of the rats.
Body weight:
All rats gained body weight throughout the observation period at all the tested doses.
Gross pathology:
No abnormality was detected at necropsy.

Any other information on results incl. tables

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)

Pre-terminal deaths

Initial

(Day 1 - at treatment)

8th  

day

Weight change

(day 8 – Initial)

15th

day

Weight change

(day 15 – Initial)

G1 and

2000

DRF

Rw4511

F

210.0

219.6

9.6

225.1

15.1

0

G1 and

2000

Main study

Rw4512

F

209.3

221.8

12.5

227.6

18.3

0

Rw4513

F

211.5

217.4

5.9

231.0

19.5

0

 Individual test item application, clinical signs, skin reactionand necropsy findings

Group & Dose

(mg/kg

body weight)

Date and time of application

Rat

Number

S

e

x

Initial

Bwt

(g)

Quantity

(mg)

applied

Observations and skin reaction

Days

1

2

3

4

5

30

min

1 h

2 h

3 h

4 h

5 h

6 h

*

Er

@

Ed

@

*

Er

@

Ed

@

*

Er

@

Ed

@

G1 and

2000

 

16 August 2019 and

10:38 AM

Rw4511

F

210.0

420

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Group & Dose

(mg/kg

body weight)

Date and time of application

Rat

Number

S

e

x

Initial

Bwt

(g)

Quantity

(mg)

applied

Observations and skin reaction

Days

1

2

3

4

5

30

min

1 h

2 h

3 h

4 h

5 h

6 h

*

Er @

Ed @

*

Er @

Ed @

*

Er @

Ed @

G1 and

2000

 

20 August 2019

10:39 AM to
10:40 AM

Rw4512

F

209.3

419

N

N

N

N

N

N

N

N

N

0

0

N

0

0

N

0

0

Rw4513

F

211.5

423

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

S

e

x

Observation

Necropsy

findings

Days

6

7

8

9

10

11

12

13

14

15

G1 and

2000

 

Rw4511

F

N

N

N

N

N

N

N

N

N

N

NAD

Group & Dose

(mg/kg

body weight)

Animal

Number

S

e

x

Observations

Necropsy

findings

Days

6

7

8

9

10

11

12

13

14

15

G1 and

2000

 

Rw4512

F

N

N

N

N

N

N

N

N

N

N

NAD

Rw4513

 

F

N

N

N

N

N

N

N

N

N

N

NAD

Legend:

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

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Conclusions:
Based on the results of the present study (acute dermal toxicity), the test item, Pyrogallol/ Pyrogallic acid are classified as follows:

The test item is classified as “Category 5” (the dermal LD50 range of 2000 < Acute Toxicity Estimates ≤ 5000 mg/kg) as per Globally Harmonized System of Classification and Labelling of Chemicals (GHS) Seventh Revised Edition, United Nations (2017). ST/SG/AC.10/30/Rev.7, as there was no mortality observed at 2000 mg/kg body weight.
Executive summary:

The acute dermal toxicity of Pyrogallol/Pyrogallic acid was tested with dose 2000 mg/kg body weight with 1 female for dose range finding study, followed by additional 2 females for main study in Wistar rats.

Based on the individual body weight, the test item at the dose of 2000 mg/kg body weight was weighed on an aluminium foil and made into a paste by adding 0.4 mL of milli-Q water and completely transferred on to the cotton gauze (size: Females: 8 x 5 cm of 6 ply) and applied directly (semi-occlusive) to the clipped skin of the rat to cover about 10% of 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.

After the 24 hours contact period, the dressing was removed, and the applied area was washed with water and wiped dry using clean towels. 

All the rats were observed for clinical signs of toxicity and mortality for 14 days post application. In addition, the site of application was observed for skin reactions at 24, 48 and 72 hours after removal of test chemical using the Draize method. Body weights were recorded on days 1, 8 and 15. All the rats gained weight during experimental period. At the end of observation period, all surviving animals were euthanized and subjected to necropsy.

There were no skin reactions and clinical signs in any of the rats. There were no abnormalities detected at the necropsy.

Based on the present study results, the acute dermal LD50of Pyrogallol/ Pyrogallic acidis more than 2000 mg/kg body weight in female Wistar rats. Test item is also considered as not irritant to the skin in female Wistar rats.

The test item,Pyrogallol\ Pyrogallic acidis classified as follows:

The test item is classified as “Category 5” (the dermal LD50range of 2000< Acute Toxicity Estimates ≤5000 mg/kg) as per Globally Harmonized System of Classification and Labelling of Chemicals (GHS) Seventh Revised Edition, United Nations (2017). ST/SG/AC.10/30/Rev.7,as there was no mortality observed at 2000mg/kg body weight.