Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Study Initiation date: September 11, 2017 and Study Completion date: November 10, 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 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Dark blue solid particulate/powder

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Species : Rat (Rattus norvegicus)
Strain : RccHan : WIST
Age/Weight at Dosing : 9 to 11 weeks, Weight (g) Minimum: 174.0, Maximum: 201.8
Source : Animal Breeding Facility, Jai Research Foundation
Total Number of Animals Used : Twelve females Female rats were nulliparous and non-pregnant
Acclimatisation Period : 6 to 11 days
Husbandry Practices
Caging : Polypropylene rat cages covered with stainless steel grid top were used.Autoclaved clean rice husk was used as the bedding material. Wooden chew blocks were provided as enrichment material.
Water Bottle : Each cage was supplied with a polypropylene water bottle with a stainless steel nozzle.
Housing : Three rat per cage
Room Sanitation : Daily: 1. Rack was cleaned with cloth, 2. Floor of experimental procedure room was swept, 3. All work tops and the floor were mopped with a disinfectant solution
Feed : Teklad Certified Global High Fiber Rat/Mice Feed manufactured by Envigo, USA.
Water : UV sterilized water filtered through Reverse Osmosis water filtration system.
Animal Room : BMR 29, Department of Toxicology
Temperature : 20 to 23°C
Relative Humidity : 49 to 67%
Air Changes : Minimum 15 air changes/hour
Photoperiod : The photoperiod was 12 hours artificial light and 12 hours darkness, light hours being 06:00 h – 18:00 h (photoperiod was maintained through automatic timer)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
The test item was found to be insoluble in RO water and 0.5% CMC while formed homogenous suspension in corn oil, so the actual dose formulation was prepared using corn oil as vehicle.
Required quantity [300 mg (for set I and II) and 2000 mg (for set III and IV)] were mixed in corn oil and the final volume was made up 10 mL for set I, set II, set III and set IV with corn oil.
Gavage solutions were prepared freshly prior to dosing on all the occasions.

Individual dose volume was adjusted according to body weight and dose level. All rats were dosed by oral gavage (day 0) using a BD 1 mL disposable syringe. Rats were fasted overnight prior to dosing and until three hours post-dosing.
Doses:
The first set (set I) of three female rats was given a single dose of 300 mg teast substance/ kg body weight. No mortality was observed
Second set (set II) of three female rats was administered with same dose level of 300 mg teast substance/ kg body weight. No mortality was observed
Third set (set III) of three female rats was administered with higher dose level of 2000 mg teast substance/ kg body weight. No mortality was observed
Fourth set (set IV) of three female rats was administered with same dose level of 2000 mg teast substance/ kg body weight. No mortality was observed
No. of animals per sex per dose:
Three
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Observed twice a day for morbidity and mortality for a period of 14 days following oral dosing.
Clinical signs were recorded once a day.
Individual body weight was recorded prior to dosing on day 0 and on days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: End of the 14 days observation period, all the rats were euthanised by carbon dioxide asphyxiation and were subjected to gross pathological examination, consisting of external examination and opening of the abdominal and thoracic cavities.

Results and discussion

Preliminary study:
The first set (set I) of three female rats was given a single dose of 300 mg test substance/kg body weight. No mortality was observed at this dose level so a second set (set II) of three female rats was administered with same dose level of 300 mg test substance /kg body weight. No mortality was observed at this dose level so a third set (set III) of three female rats was administered with higher dose level of 2000 mg test substance /kg body weight. No mortality was observed at this dose level so a fourth set (set IV) of three female rats was administered with same dose level of 2000 mg test substance /kg body weight. No mortality was observed at this dose level hence the endpoint was achieved and further testing was not required.
Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No mortality was observed in rats treated with 300 and 2000 mg the test substance/kg body weight.
Clinical signs:
No adverse clinical sign was observed in all the rats treated with 300 and 2000 mg the test substance/kg body weight.
Body weight:
Normal gain in body weight was observed in all the rats treated with 300 and 2000 mg the test substance/kg body weight.
Gross pathology:
External: No abnormality detected
Internal: No abnormality detected
Other findings:
- Organ weights:
- Histopathology: No abnormality detected
- Potential target organs: No abnormality detected
- Other observations: No abnormality detected

Applicant's summary and conclusion

Interpretation of results:
other: Category 5 or Unclassified
Conclusions:
No mortality was observed in rats treated with 300 and 2000 mg test substance/kg body weight. The acute oral LD50 (cut-off value) of the test substance in Wistar rats was found to be 5000 mg/kg body weight.
The acute oral median lethal dose (LD50 cut- off value) the test substance in Wistar rats was found to be 5000 mg/kg body weight.
Based on the results of this study, an indication of the classification for the test substance is ; Category 5 or Unclassified, based on Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2017)
Executive summary:

In an acute oral toxicity study, four sets of fasted Wistar rats (3 females/set) (9 to 11 weeks) were given a single oral dose ofthe test substanceat 300 (for set Iand II) and 2000 (for set III and IV) mg/kg body weight and all rats were observed for 14 days. There were no treatment-related mortality, clinical sign and changes in body weight or necropsy findings observed. The acute oral median lethal dose (LD50 cut-off value) of the test substance in Wistar rats was found to be 5000 mg/kg body weight. Based on the results of this study, an indication of the classification forthe test substanceis as follows:

Globally Harmonized System of Classification and Labelling of Chemicals (GHS 2017) : Category 5 or Unclassified