Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
February 09 - April 16, 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): CH-CA
- Appearance: transparant liquid
- Storage condition of test material: airtight at room temperature
- Stability under storage conditions: no data

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: ORIENTBIO INC., 143-1 Sangdaewon-dong, Joongwon-gu, Seongnam-si, Gyeonnggi-do, Korea
- Age at study initiation: Young adult animals (8 weeks old)
- Weight at study initiation: 176.94-190.81 g
- Housing: Individual 1 animal per cage in labeled stainless wire cages (height 21 cm)
- Diet (e.g. ad libitum): Free access to pelleted rodent diet (TEKLAD CERTIFIED GLOBAL 18% PROTEIN RODENT DIET 2918C, lot no.: 2918C 100306MA, Harlan TEKLAD, Madison, USA).
- Water (e.g. ad libitum): Free access to tap water.
- Acclimation period: The acclimatization period was 7 days before the start of treatment under laboratory conditions.
- Health inspection: A health inspection was performed prior to treatment, to ensure that the animals were in a good state of health.
- Rats were fasted for approximately 16 hours prior to dosing of the test substance, but drinking water was not withheld. The diet was provided 4 hours after the completion of the administration.

The diet was analyzed periodically by manufacturer to quantify nutrient contents and contaminants. There were no known contaminants in the diet at levels that would be expected to interfere with or affect the results of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.7 – 24.4
- Humidity (%): 32.4 - 58.0
- Sanitation and sterilization of cages, fodder tub, water bottles and animal room:
Cages, fodder tub and water bottles were washed by the cage washer using Ultra-Clean Neighbour-PC (alkali surfactant, NE-SARANG Co., Ltd., Korea) and Ultra-Clean Neighbour-RM300 (surfactant, NE-SARANG Co., Ltd., Korea) and they were sterilized by an autoclave. Cages, fodder tub and water bottles were replaced after acclimation period.
After the completion of daily work, animal room was sanitized using disinfect solutions such as Pacom - A (component: Octyldecyl dimethyl ammonium chloride, 200 fold, SAMU MEDIAN Co., Ltd., Kora), MICRO-QUAT (component: Alkyldimethyl-benzyl ammonium chloride, 400 fold, ECOLAB Co., Ltd., Korea) and BROMO-SEPT (component : Di-decyl di-methyl ammonium bromide, 300 fold, DAESUNG MICROBIOLOGICAL LABS. Co., Ltd., Korea) which were changed every 10 days, respectively.
- Air changes (per hr): approximately 10-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: February 26 and february 28, 2007

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Justification for choice of vehicle: Corn oil was chosen as a vehicle since it was confirmed that the test substance was dissolved into the vehicle through the pre-preparation of the test substance.

MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg body weight.

DOSAGE PREPARATION: For dosing formulation, the necessary amount of the test substance was weighed and dissolved in the vehicle. The dosing formulation was mixed with a vortex mixer until in solution. Dosing formulations were prepared on each day of dosing step 1 and 2.

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/Viability and Clinical signs: The animals were observed continuous during the 30 minutes, at 1, 2, 4 and 6 hours after dosing and subsequently once daily up to 14 days (from Day 2 to 15).
Body weights: Body weights were measured prior to dosing (Day 1), at Day 4, 8 and the day of necropsy (day 15) with the automatic animal balance.
- Necropsy of survivors performed: Yes
Statistics:
Average values were calculated with measured values obtained through this study and statistical analysis of data related with this study was not 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:
During the observation period, there were no deaths for all animals in step 1 and 2.
Clinical signs:
There were no abnormal clinical signs noted for all animals in step 1 and 2.
Body weight:
Body weights were normally increased in all animals over the study period.
Gross pathology:
In the necropsy of step 1 and 2, there were no macroscopic findings on external surface and organs of body cavity (cranial, thoracic and abdominal).

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 performed according to OECD 423 and GLP, the LD50 value of CH-CA in Sprague-Dawley rats was established to exceed 2000 mg/kg body weight.