Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
This study was conducted according to the following Good Laboratory Practices: - “OECD Principles of Good Laboratory Practice” Organisation for Economic Co-operation and Development, ENV/MC/CHEM (98)17 (as revised in 1997) This study was conducted according to the following test guideline: - “OECD Guideline for Testing of Chemicals 423, Acute Oral Toxicity-Acute Toxic Class Method” Organisation for Economic Co-operation and Development (Adopted: 17th December 2001)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
2.3.1 Species, Strain Rat, Sprague-Dawley (Crl:CD(SD)), SPF
2.3.2 Producer & supplier ORIENTBIO INC., Korea
2.3.3 Justification for species selection
Sprague-Dawley rats are commonly used in toxicity studies with a large historical control data base.
2.3.4 Sex, number, age and the body weight range (at receipt)
Females, 13 rats, 7 weeks old, 166.0–176.7 g
2.3.5 Sex, number, age and the body weight range (on administration)
Females, 12 rats, 8–9 weeks old, 173.9–214.0 g

2.4.1 Quarantine room No. A315
2.4.2 Animal room No. A323
2.4.3 Type & size of a cage Stainless wire mesh cage, 260W×350D×210H (mm)
2.4.4 Number of animals per cage One animal/cage (during the study)
2.4.5 Temperature 21.0–24.4°C
2.4.6 Relative humidity 43.3–62.9%
2.4.7 Air changes 10–15 clean, fresh, filtered air changes per hour
2.4.8 Lighting 12 hour light / dark cycle
(7 AM to 7 PM via automated timer)
2.4.9 Intensity of illumination 150–300 Lux
2.4.10 Cage washing Every two weeks
Cages and feeders were washed in the cage washer and sterilized by an autoclave.

Administration / exposure

Route of administration:
other: gastric intubation
Vehicle:
water
Details on oral exposure:
2.7.1 Route
Oral via gastric intubation
2.7.2 Justification for the route of administration
The oral route was chosen to assess the toxicity by oral exposure of the test substance.
2.7.3 Method of administration
Individual doses were calculated based on the animals body weight recorded just prior to dosing at a dose volume of 10 mL/kg body weight. Animals were dosed via gastric intubation with 3 mL disposable syringes fitted with intubation tubes. Animals were fasted overnight, approximately 16 hours prior to dosing. Drinking water was provided ad libitum. Feed was provided approximately 4 hours post dosing.
Doses:
300mg/kg, 2000mg/kg
No. of animals per sex per dose:
3F
Control animals:
no
Details on study design:
The starting dose level for this study was selected at 300 mg/kg, since there was no
available information on the toxicity of the test substance.
The following sequential dosing steps were based on mortality and clinical observations for 2 to 5 days after the previous dose level. The following dosage regimen was selected according to the ‘ ANNEX 2c : TEST PROCEDURE WITH A STARTING DOSE OF 300 MG/KG BODY WEIGHT’.

2.9 Parameters Evaluated
2.9.1 Clinical signs
All animals were observed for mortality, general condition and clinical signs (time, onset, severity and recovery) 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 (Days 1 to 14).
2.9.2 Disposition of dead animal
During the observation period, necropsy was done on one animal found dead immediately after body weight measurement.
2.9.3 Body weights
Body weights were recorded prior to dosing (Day 0), on Days 1, 3 and 7 and on the day of necropsy, Day 14.
2.9.4 Necropsy
On Day 14, all surviving animals were anesthetized with CO2 and exsanguinated from the abdominal aorta. Complete gross postmortem examinations were performed on all animals in the study.
2.9.5 Histopathology
Since no gross findings were observed at necropsy, histopathological examinations were not performed at 300 mg/kg.
Histopathological evaluations were performed on the forelimb, jejunum, skin and/or stomach. There was a visible indication of morphologic abnormalities in one dead animal and two surviving animals at 2,000 mg/kg.
Statistics:
Statistical analysis was not performed. Mean scores and values are presented.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
All animals at 300 mg/kg survived the duration of the study. There were no effects on the mortality.
One animal at 2,000 mg/kg was found dead on Day 2 after dosing. It was considered to be under the effect of the test substance.
Clinical signs:
There was no death in any animals at 300 mg/kg. Irregular respiration was evident in all animals at 300 mg/kg on the day of dosing and on Day 1 after dosing. These animals returned to a normal appearance on Day 2 after dosing.
In one dead dosed with 2,000 mg/kg, irregular respiration and compound-colored stool were evident on the day of dosing and on Day 1 after dosing. Irregular respiration decrease of fecal volume, hypothermia, incised wound (left forelimb, right forelimb), paleness, refusal to feed, scratched wound (abdomen), self biting and in a state of lying on side were evident on Day 2 after dosing. Then, one animal was found dead on Day 2 after dosing. In five surviving animals at 2,000 mg/kg, irregular respiration was evident on the day of dosing. In three surviving animals at 2,000 mg/kg, irregular respiration and/or compound-colored stool were evident from Day 1 to Day 2 after dosing. Then, these animals returned to a normal appearance on Day 3 after dosing. In two surviving animals at 2,000 mg/kg, irregular respiration, compound-colored stool, decrease of fecal volume, hypothermia, incised wound (left forelimb, right forelimb), paleness, decrease in food consumption, blackish stool, piloerection and/or self biting were evident from Day 1 to Day 6 after dosing. Crust formation (left forelimb, right forelimb) was evident from Day 3 to Day 14 after dosing.
Body weight:
Normal body weight gain was evident in all animals at 300 mg/kg.
A tendency to suppress body weight gain was evident in all surviving animals on Day 1 and/or on Day 3. The decrease in body weight was evident in four animals on Day 3 after dosing at 2,000 mg/kg. Then, five animals returned to normal on Day 7. These changes were considered to be test substance-related effects.
Gross pathology:
No remarkable findings were noted in all remaining females at 2,000 mg/kg.

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Remarks:
Migrated information
Conclusions:
Based on the results of the acute oral toxicity study in Sprague-Dawley rats, GHS classification of the test substance, CMS Red 620, was classified to be Category 5.