Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2015-03-24 to 2015-04-14
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study

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:
adopted 2001-12-17
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:
- Physical state: colourless to light yellow, clear liquid
- Storage condition of test material: room temperature (measurement value: 20.8 to 22.2°C, permissible range: 15 to 25°C)

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS - Crl:CD(SD)
- Source: ORIENTBIO INC., Korea
- Age: 8 week old
- Weight: 173.6 to 189.5 g
- Fasting period before study: animals were fasted overnight, approximately 16 hours prior to dosing. feed was provided approximately 4 hours post dosing.
- Housing: stainless wire mesh cage (260W x 350D x 210H (mm)); one animal/cage
- Diet (ad libitum, except for a fasting period before the study as described above): pelleted rodent chow (Teklad Certified Irradiated Global 18% Portein Rodent Diet 2918C; Harlan Laboratories, Inc., U.S.A.))
- Water (ad libitum): public tap water in Cheongju-si was filtered and irradiated by ultraviolet light
- Quarantine/Acclimation period: 3 day / 4 days

ENVIRONMENTAL CONDITIONS
- Temperature: 21.0 to 23.8°C
- Relative humidity: 42.2 to 64.8%
- Air changes: 10 to 15 clean, fresh, filtered air changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Lot no.: MKBP7039V

MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw
Individual doses were calculated based on the animals' body weight recorded just prior to dosing.

DOSAGE PREPARATION:
The required amount of the test substance was weighed on an electronic balance and placed in a bottle. A small amount of vehicle was added and mixed using a vortex mixer until dissolved. The vehicle was gradually added to yield the desired concentration (400 mg/mL). All preparations were conducted just prior to use.

CLASS METHOD
- Rationale for the selection of the starting dose: due to the low expected toxicity of the test substance, based on information supplied by the sponsor, 2000 mg/kg bw was selected as the starting dose for this study.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 female rats (3 female rats per dosing step)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: all animals were observed at 30 minutes after dosing and at 1, 2, 4 and 6 hours after dosing on Day 0 and once daily thereafter for 14 days.
Body weights were recorded prior to dosing (Day 0), on Days 1, 3 and 7 and on the day of necropsy, Day 14.
- Necropsy of survivors performed: yes, all surviving animales were sacrificed on day 14. Complete gross postmortem examinations were performed on all animals.
Statistics:
not applicable

Results and discussion

Effect levelsopen allclose all
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
female
Dose descriptor:
LD50
Effect level:
>= 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: LD50 cut-off value based on the flow chart according to the OECD guideline 423 .
Mortality:
All animals dosed at 2000 mg/kg survived the duration of the study.
Clinical signs:
Lacrimation and/or salivation were evident in five animals on the day of dosing. Mucous stool and/or soiled perineal region were observed in 3 animals on Day 1, but they returned to a normal appearance on Day 2. One rat presented clinical signs and those were decreased fecal volume or no stool from Day 1 to Day 3, refusal to feed on Day 2 only, decrease in locomotor activity and irregular respiration from Day 2 until Day 4, but all clinical signs returned to normal from Day 5 onwards.
These clinical signs were considered to be test substance-related effects.
Body weight:
A decrease in body weight or a tendency to suppress body weight was evident in five animals on Day 1 and Day 3. Then, five animals retunred to normal appearance on Day 7. These changes were considered to be test substance-related effects.
Gross pathology:
No grossly visible evidence of morphologic abnormalities was evident in any animals.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
LD50 cut-off value (female rats) ≥ 5000 mg/kg bw
According to the Directive 67/548/EEC and its subsequent amendments, the test substance is not acutely toxic via the oral route.
According to the Regulation (EC) No 1272/2008 and subsequent regulations, the test item is not acutely toxic via the oral route.
Executive summary:

The acute oral toxicity of the test substance was investigated according to the OECD guideline 423 (2001). Two dose groups of three females received a single dose of 2000 mg/kg bw in corn oil via gavage. Clinical signs, mortality, and body weight were recorded. All animals were subjected to gross necropsy.

No mortality was observed during the study. Lacrimation and/or salivation were evident in five animals on the day of dosing. Mucous stool and/or soiled perineal region were observed in 3 animals on Day 1, but they returned to a normal appearance on Day 2. One rat presented clinical signs and those were decreased fecal volume or no stool from Day 1 to Day 3, refusal to feed on Day 2 only, decrease in locomotor activity and irregular respiration from Day 2 until Day 4, but all clinical signs returned to normal from Day 5 onwards. These clinical signs were considered to be test substance-related effects. Furthermore, a decrease in body weight or a tendency to suppress body weight was evident in five animals on Day 1 and Day 3. The five animals retunred to normal appearance on Day 7. These changes were considered to be test substance-related effects. Lastly, no grossly visible evidence of morphologic abnormalities was evident in any animals.

Based on these results the LD50 cut-off value was considered to be equal or greater than 5000 mg/kg bw for female rats.

According to the Directive 67/548/EEC and its subsequent amendments, the test substance is not acutely toxic via the oral route.

According to the Regulation (EC) No 1272/2008 and subsequent regulations, the test item is not acutely toxic via the oral route.