Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
4 March 2019 (Study Initiation) to XX 2019 (Study Completion)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

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)
Deviations:
no
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
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material: Nitika Pharmaceutical Specialities Pvt. Ltd
- Lot/batch No.of test material: IRST7H780L
- Expiration date of the lot/batch: August 2023
- Purity test (release) date: September 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (ambient), container kept tightly closed in a dry, cool and well ventilated place
- Stability under test conditions: Assumed stable for the duration of the study
- Solubility and stability of the test substance in the solvent/vehicle: solubility not applicable. The required amount of test item was applied as received (moistened with Reverse Osmosis water) over the clipped dorsolateral thoracic surface of the skin (approximately 7 cm × 5 cm body surface area). The test item contact period with the skin was 24 hours, rats were thereafter observed for a period of 14 days.

- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None,

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none, test item administered as supplied (moistened with Reverse Osmosis water) before application

OTHER SPECIFICS:
- measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical is added: The test item pH (1% solution in distilled water at room temperature) was checked and found to be acceptable at 6.96 ± 0.04.

Test animals

Species:
rat
Strain:
Wistar
Remarks:
RccHan
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Animal Breeding Facility, Jai Research Foundation, India.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 12 to 14 weeks
- Weight at study initiation: Minimum: 243.7, Maximum: 287.3
- Fasting period before study: None
- Housing: Polypropylene rat cages covered with stainless steel grid tops. Autoclaved clean rice husk as bedding material. Wooden blocks were provided as enrichment material.
- Diet: ad libitum) Teklad certified Global High Fiber Rat/Mice Feed manufactured by Envigo, USA.
- Water: ad libitum , UV sterilized water filtered through a Reverse Osmosis (RO) water filtration system.
- Acclimation period: 7 to 19 days

ENVIRONMENTAL CONDITIONS acclim
- Temperature (°C): 20 to 23 °C
- Humidity (%): 57 to 67%
- Air changes (per hr): Minimum of 15/hour
- Photoperiod (hrs dark / hrs light): The photoperiod was maintained through automatic timer (12 h artificial light and 12 h darkness), light hours being 06:00 – 18:00 h.

IN-LIFE DATES: From: To: 9 March 2019 (Experimental Start) to 11 April (2019)

Administration / exposure

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Remarks:
moistened prior to application with RO water
Details on dermal exposure:
TEST SITE
- Area of exposure: The fur of each rat was closely clipped from the dorsal area of the trunk using an electric clipper without abrading or damaging the skin. An area greater than 10 percent of the body surface area was clipped approximately 24 h prior to test item application.
- % coverage: An area greater than 10 percent of the body surface area was clipped approximately 24 h prior to test item application.
- Type of wrap if used: The test item was held in contact with the skin using porous gauze dressing (not more than 8 ply) and a non-irritating tape throughout the 24 h exposure period to prevent any loss of test item and also to ensure that the rats did not lick or ingest test item.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): At the end of the exposure period, any residual test item was removed using cotton soaked in RO water.
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (weight with unit): Based on individual body weights, the required amount of test item was applied (moistened with RO water).
- For solids, paste formed: no

VEHICLE
Not applicable, test item administered as supplied (moistened with RO water) over the clipped dorsolateral thoracic surface of the skin.
Duration of exposure:
24h
Doses:
Range finding study: 200, 1000 and 2000 mg/kg bw
Main study: 2000 mg/kg bw
No. of animals per sex per dose:
1 for each dose range finding concentration
2 for the main study at 2000 mg/kg bw
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Rats were observed for signs of toxicity and mortality within 0.5 hours and at 2, 4 and 6 h post-dermal application on the day of dosing (day 0). Subsequently, rats were observed twice a day for morbidity and mortality for a period of 14 days following dermal application. Rats were observed for erythema and oedema at 24 h, 48 h and 72 h post patch removal. The clinical signs were recorded once a day. Individual body weight was recorded prior to dermal application on day 0 and on days 7 and 14 post dermal application.

- Necropsy of survivors performed: yes, all rats at the end of the 14-day observation period were euthanised by carbon dioxide asphyxiation and subjected to a gross pathological examination consisting of an external examination and opening of abdominal and thoracic cavities.

- Other examinations performed: clinical observations and body weight.
Statistics:
None

Results and discussion

Preliminary study:
As no toxicological information was available for the test item, a range finding study was conducted. Initially, rat N˚ 1 was dosed at 200 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg body weight. As no mortality was observed after 72 h exposure, rat N˚ 2 was dosed at 1000 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg body weight. As no mortality was observed after 72 h exposure, rat N˚ 3 was dosed at the limit guideline dose of 2000 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg body weight.
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:
No morbidity and mortality was observed in any rat treated with 200, 1000 or 2000 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg bw.
Clinical signs:
No sign of toxicity was observed in any rat treated with 200, 1000 or 2000 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg bw.
No erythema or oedema was observed at any 24 h time point interval (24 h, 48 h, and 72 h) post patch removal in any rat.
Body weight:
Normal gain in body weight was observed in all rats treated with 200, 1000 or 2000 mg Fatty acids, C16-18 (even numbered), iron(III) salts/kg bw.
Gross pathology:
Necropsy (Macroscopic Findings)
External examination of terminally sacrificed rats did not reveal any abnormality.
Internal (Visceral) examination of terminally sacrificed rats did not reveal any abnormality.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
As per the Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2017), if the test item is classified under Category 5 or Unclassified it indicates that the LD50 is established as greater than 2000 mg/kg body weight.