Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from experimental study report.

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 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
This study was designed to determine the dermal LD50 of the test item (up to 2000 mg/kg) or to establish a non-lethal dose level of 2000 milligram of test item per kilogram of body weight.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report): Sodium 2-amino-4, 6-dinitrophenoxide
- Molecular formula : C6H4N3NaO5
- Molecular weight : 221.1036 g/mol
- Smiles notation: [Na+].Nc1cc(cc(c1[O-])[N+](=O)[O-])[N+](=O)[O-]
- InChl : 1S/C6H5N3O5.Na/c7-4-1-3(8(11)12)2-5(6(4)10)9(13)14;/h1-2,10H,7H2;/q;+1/p-1
- Substance type: Organic
- Physical state: Solid powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult (8 to 10 weeks old) female rats were used.
- Weight at study initiation: The weight range of approximately 221.5 to 227.4 grams at initiation of dosing.
Body weights at the start : Female Mean: 225.23 g (= 100 %); Minimum : 221.5 g (- 1.66 %); Maximum : 227.4 g (+ 0.96 %)
- Identification: Each rat was individually identified by the cage number.
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period:at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 to 21.8 degree centigrade
- Humidity (%): 50.4% to 60.3%.
- Air changes (per hr): The animal room was independently provided with at least ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: From: 07-05-2018 To: 17-06-2018

Administration / exposure

Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: the trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: approximately 10% of the total body surface area
- Type of wrap if used: The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.
- Time after start of exposure: 24 hours
Duration of exposure:
24 hours
Doses:
Dose Range Finding Study: Group I : 2000 mg/kg
Main Study: Group II : 2000 mg/kg
No. of animals per sex per dose:
Dose Range Finding Study: Group I : 2000 mg/kg - 1
Main Study: Group II : 2000 mg/kg - 2
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: yes, necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).
- Other examinations performed: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality, until sacrifice. Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.
Evaluation of Dermal Reaction: Dermal reaction was observed daily for study period of 14 days.
Body weights: Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.
Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.
Statistics:
not specified

Results and discussion

Preliminary study:
Dose Range Finding Study: Based on the available literature a single dose of 2000 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity were observed during first 48 hours.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Dose Range Finding Study: Group I : Animal treated at the dose level of 2000 mg/kg body weight survived through the study period of 14 days.
Main Study: Group II : Animals treated at the dose level of 2000 mg/kg body weight survived through the study period of 14 days.
Clinical signs:
Dose Range Finding Study: Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.
Main Study: Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.
Body weight:
Dose Range Finding Study: Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 8.35% and 15.85% respectively.
Main Study: Group II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.88% and 12.57% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group from dose range finding study and main study sacrificed terminally.
Other findings:
Evaluation of Dermal Reaction - Dose Range Finding Study: Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Main Study: Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Any other information on results incl. tables

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal No.

Period of signs in days

 From - to

 

Mortality

I

2000

No clinical signs observed

1

1

Day 0 - Day 14

0/1

 

Main Study:

Group

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

 From - to

 

Mortality

II

2000

No clinical signs observed

2

2, 3

Day 0 - Day 14

0/2

 

Table No. II 

Summary of Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group

No.

Dose mg/kg

Dermal Reaction

Total Number of

Animals

Animal

No.

Period of signs

in days

 From – to

I

2000

No dermal reaction observed

1

1

Day 0 - Day 14

 

Main Study:

Group

No.

Dose mg/kg

Dermal Reaction

Total Number of

Animals

Animal Nos.

Period of signs

in days

 From – to

II

2000

No dermal reaction observed

2

2, 3

Day 0 - Day 14

  

Table No.III

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group No.

Dose

(mg/kg body weight)

 

Body weight Day 0

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

2000

Mean

221.50

240.00

8.35

256.60

6.92

15.85

± SD

-

-

-

-

-

-

 

Main Study:

Group No.

Dose

(mg/kg body weight)

 

Body weight Day 0

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

II

2000

Mean

227.10

240.45

5.88

255.65

6.32

12.57

± SD

0.42

0.07

0.23

0.49

0.17

0.43

 

 

Table No.IV

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group No.

Dose

mg/kg

Animal Number

Animal Fate

Gross Pathological Findings

I

2000

1

TS

No abnormality detected

 

                    Main Study:

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

2000

2, 3

TS

No abnormality detected

 

                     TS = Terminal Sacrifice

Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.
Executive summary:

The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

In the dose range finding study a single dose of 2000 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity were observed during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs of toxicity at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals.  The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals. Animals from dose range finding study and main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Under the condition of the study, it was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.