Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
07 April, 2015 - 24 April, 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
(2001)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): IN-1000
- Appearance: Brown, opaque liquid
- Storage condition of test material: Refrigerated (2ºC to 8ºC) in a closed container and under nitrogen

Test animals

Species:
rat
Strain:
other: Crl:CD(SD)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Inc., Raleigh, NC
- Age at study initiation: Young adult animals (approx. 8 weeks old)
- Weight at study initiation: 171 to 184 g, ± 20% of the mean.
- Fasting period before study: Overnight prior to dosing
- Housing: Housed individually in clean, stainless steel, wire-mesh cages suspended above cage-board.
- Diet: Free access to basal diet (PMI Nutrition International, LLC, Certified Rodent LabDiet® 5002)
- Water: Free access to municipal water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.3 – 21.4
- Humidity (%): 39.1 - 58.4
- Air changes (per hr): minimum 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

MAXIMUM DOSE VOLUME APPLIED: 1.33 mL/kg body weight
The test substance was dosed undiluted based on its density (tested as received, no correction for the purity). The dose volume was determined by dividing each dose level, expressed as g/kg bw, by the density (1.503 g/mL, as determined by WIL Research Formulations Department personnel). Individual doses were calculated based on fasted body weights taken just prior to dosing.

DOSAGE PREPARATION: The following steps were performed in a glove box under nitrogen. Prior to use, the bulk test substance container was inverted/swirled. A sufficient amount of test substance (pH 5, litmus paper) was transferred into an amber glass vial and a stir bar was added.
The container was purged with nitrogen and capped immediately. The contents were stirred continuously throughout use.

Doses:
2000 mg/kg body weight

No. of animals per sex per dose:
6 (2 groups of three females in a stepwise manner)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality: The rats were observed at approximately 15 minutes (± 5 minutes) and 1, 2 and 4 hours post-dosing on study day 0 and twice daily, once in the morning and once in the afternoon, thereafter for 14 days.
Body weights: Body weights were obtained and recorded on study days 0 (initiation), 7, and 14 (termination).
Clinical observations: The rats were observed at approximately 15 minutes (± 5 minutes) and 1, 2, and 4 hours post-dosing on study day 0 and once daily thereafter for 14 days.
- Necropsy of survivors performed: On study day 14, the rats were euthanized by carbon dioxide inhalation. The major organ systems of the cranial, thoracic, and abdominal cavities and the eyes and skin were examined for all animals.
- Other examinations performed: none.
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
All animals survived to the scheduled necropsy (study day 14).
Clinical signs:
There were no test substance-related clinical observations. Clinical observations were limited to yellow material on the anogenital area and hindlimb(s) for 1 female on study day 1.
Body weight:
There were no remarkable body weight changes noted during the study.
Gross pathology:
There were no macroscopic findings at the scheduled necropsy.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
In an acute oral toxicity study with rats, performed according to OECD 423 test guideline and GLP principles, an LD50 >2000 mg/kg bw was determined for IN-1000.



Executive summary:

The acute oral toxicity of IN-1000 was determined in accordance with OECD 423 guideline and according to GLP principles.

IN-000 was administered to six female rats by a single dose of 2000 mg/kg bodyweight (2 groups of three females in a stepwise manner). All animals survived to the scheduled necropsy (study day 14). There were no test substance-related clinical observations (clinical observations were limited to yellow material on the anogenital area and hindlimb(s) for 1 female on study day 1). There were no remarkable body weight changes noted during the study and there were no macroscopic findings at the scheduled necropsy.

Based on the results of this study, the estimated LD50 of IN-1000 was greater than 2000 mg/kg body weight and the substance does not need to be classified for acute oral toxicity according to Regulation (EC) No 1272/2008 on classification, labelling and packaging.