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Description of key information

The test substance was found to have an LD50 of 300< LD50 ≤2000 mg/kg in the acute oral toxicity study. The substance is classified in Category 4 for Acute Oral Toxicity, in accordance with EU CLP criteria. The LD50 found in the acute dermal toxicity study was >2500 mg/kg bw, as such EMI-DBS is not classified as an acute dermal toxin in accordance with EU CLP criteria.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
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: Study was conducted by a GLP accredited laboratory using OECD Testing Guideline 423.
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
Species and strain: Specific Pathogen Free (SPF) Sprague Dawley (SD) rats
Supplier and producer: ORIENT BIO INC. (Korea)
Reasons for choice of the species: SD rats have been applied widely in general toxicity tests as a suitable experimental animal for toxicity testing. In addition, sufficient raw data has been accumulated and is available for interpretation and evaluation of study results.
Date of acquisition: 1st acquisition: 20 October 2011, 2nd acquisition: 3 November 2011.
Number of animals received: 7 female rats for each acquisition
Age of animals received: 7 weeks
Body weight on arrival: 1st: 173.12-180.57 g, 2nd: 169.61-194.22 g.
Quarantine and acclimation: Animals were acclimated for 7-14 days. Only animals with the best appearance were selected for the test after observation during the acclimation period, animals were accepted based on the certification provided by the supplier.
Age at the intiation of the administration: 8-9 weeks
Body weight at the administration: 1st step: 300 mg/kg: 189.37-195.99, 2nd step: 300 mg/kg: 198.61-212.87 g, 3rd step: 2000 mg/kg: 197.44-206.76 kg.
Number of animals administered test substance: Each step used 3 female rats. Total 9 female rats were administered the test substance.
Grouping: Animals were weighed one day before the test substance administration, and grouped to ensure a distribution of graded body weight.
Identification of animals: Individual animals were identified by tail marking with an oily-ink felt pen. Individual cages were distinguished by the individual card labelling. The record sheets provided at the entrance of the SPF amimal room contained the study number, the study title, the duration of the SPFroom use, the name of the study director and the names of the study personnel.
Disposal of remaining animals: Remaining animals were euthanized.
Animal care room: Room 1 in the SPF animal facility area.
Temperature and humidity during test work: 22.7 ± 0.8°C and 53.0 ± 5.1 %RH.
Ventilation frequency: 10-15 air changes/hour
Lighting cycle: 12 hours duration (lighting on at 8 am and off at 8 pm)
Lighting intensity: 281 Lux
Ambient noise level: 52.8 dB
Ammonia concentration: less than 5 ppm
Housing: All animals were housed in a wire mesh cage (250 W x 350 L x 180 H mm) during quarantine, acclimation, administration and observation period.
Animal number per cage: During the experiment, not more than 3 animals were housed in a cage.
Change of cages: At grouping, test animal cages were changed.
Feed: (1) Tyep: Radiation sterilized, solid laboratory animal feed (2): Supplier: DooYeol Biotech Co., Ltd (3) Producer: Harlan Laboratories, Inc., USA (4) Supply Method: ad libitum (5) Analysis: Feed analysis data was provided from the supplier
Water: (1) Type: Incheon, Korea municipal tap water purified by reverse osmosis filtering system (2) Supplying method: ad libitum using water bottles (3) Analysis: Performed by national certificated inspection organisation.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
Vehicle Information

Name: Corn oil
Lot No: MKBF2096V
CAS No: 8001-30-7
Received Date: 23 June 2011
Received Quantity: 500 ml
Appearance: Pale yellow liquid
Storage Condition: At room temperature
Manufacturer: SIGMA
Justification for choice: According to the solubility test, prior to study initiation, corn oil was chosen because the test substance solution suspended evenly in corn oil by heating (approximately at 40°C) and shaking (sonication) for 10 minutes.

Preparation of the test substance

The test substance was measured on the administration day, prepared and suspended in corn oil. For the first and second step, 0.30 g of the test substance was suspended in corn oil by heating (approximately at 40°C) and shaking (sonication) for 10 minutes to make a total volume of 10 ml at the dose level of 300 mg/kg. For the third step, 2 g of the test substance was suspended in corn oil by heating (approximately at 40°C) and shaking (sonication) for 10 minutes to make a total volume of 10 ml at the dose level of 2000 mg/kg. To clarify the homogeneity of test samples, a homogeneity test was performed with HPLC (MAI-072-01). In the test result, it was found that the standard curve had a linearity at 25 µg/ml - 100 µg/ml concentrations and these data emerged reproducible. The coefficient of variation for the EMI-DBS is 1.91%. According to homogeneity analysis result, it was confirmed that the test samples had homogeneity.
Doses:
The dose level was determined in accordance with 'Acute Toxic Class Method' the Test Guidelines of the National Institute of Environment Research (NIER) [Notice No. 2010-29, (revised 16 August 2010)] and the OECD Guideline for Testing of Chemicals No. 423 'Acute Toxic Class Method' (Adopted 17 December 2001). The first step dose was 300 mg/kg due to an absence of toxicity data of the test substance. The second step dose was 300 mg/kg and the third step dose was 2000 mg/kg.
No. of animals per sex per dose:
3 female rats per dose.
Control animals:
no
Details on study design:
Route of administration: Oral
Reason for choice of the route of administration: Common method to evaluate acute oral toxicity
Method of administration: Rats were fasted one night before the administration day to empty their stomach. An intubation cannula was used for the oral administration. All fasted test animals were fed approximatelt at three to four hours after test substance administration.
Calculation of dosing volume: Individual dosage was adjusted based on fasted body weight measured right before the administration. Dosing volume was 10 ml/kg.
Frequency and period of administration: Once

Observations

(1) Clinical signs and mortalities: General clinical signs or mortalities of all treated animals were observed continuously during the first half-hour and one hour from the administration time. After that, the animals were observed once hourly up to the first six hours on the administration day. From the next day, each animal was observed once every day up to 14 days after the administration.
(2) Body weight: Individual animals were weighed before the administration and at 1, 4, 7 and 14 days after the administration.
(3) Necropsy and gross findings examination: On day 14 after the administration, all surviving animals were anesthetized with CO2 gas, and terminated by exsanguination from the aorta. Complete post-mortem examinations were performed on all vital organs.
Statistics:
Data analysis

Each body weight change was evaluated during the observation period. The mean value and the standard deviation of each animal group were applied to resulting data. No additional statistical assessment was performed in data analysis. The LD50 value was classified according to the Notice No. 2010-29 (revised 16 August 2010) of the National Institute of Environmental Research.
Sex:
female
Dose descriptor:
LD50
Effect level:
300 - <= 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No 95% CL were provided
Mortality:
No mortalities were observed during the observation period in the first step (300 mg/kg) and the second step (300 mg/kg). There were two dead animals (animal number 7 and 8) 1 day after administration in the third step (2000 mg/kg).
Clinical signs:
No unusual clinical signs were observed during the observation period in the first step (300 mg/kg) and the second step (300 mg/kg). In the third step (2000 mg/kg) clinical signs were observed. These were mucous stool and inanimation from 2 to 6 hours after administration.
Body weight:
A decrease in body weight was seen in one animal (animal number 1) in the first step 4 days after administration, compared with the body weight 1 day after administration. In the other animals there were normal body weight gains. In the second step, there was a decrease in the body weight of one animal (animal number 5) 7 days after administration compare with the body weight 4 days after administration. In other animals, there were normal body weight gains. In the third step, one animal (animal number 9) was found to have a decreased body weight 1 day after administration, compared with body weight before administration. There were normal body weight gains after that time.
Gross pathology:
No abnormal gross findings were observed during necropsy on all animals in the first and second steps. In the third step, no gross abnormalities were found in necropsy of the surviving animals or the dead animals.
Interpretation of results:
Toxicity Category IV
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the results of the of the acute oral toxicity study of EMI-DBS to Sprague-Dawley rats, the lethal dose 50 (LD50) is considered to be GHS classification 4 (300< LD50 ≤2000 mg/kg) in this study.
Executive summary:

The acute oral toxicity of the test substance was determined in accordance with the OECD Guideline for Testing of Chemicals 423. The test substance was orally administered to female Sprague-Dawley rats. The first and second step doses were 300 mg/kg bw/day, the third step dose was 2000 mg/kg bw/day. The first step dose was chosen due to an absence of toxicity data on the test substance. Mortalities, clinical signs and body weight changes were monitored during the 14 -day observation period. Necropsy and gross findings were performed for all animals. There were no dead animals or unusal clinical signs or unusual observations at necropsy in the animals in the first and second steps (300 mg/kg). There were two dead animals 1 day after administration of the test substance in the third step (2000 mg/kg). Mucous stool and inanimation on the administration day were recorded as clinical signs in the third step. No abnormal gross findings were observed during necropsy. Based on the results of the study, EMI-DBS was found to have an LD50 of 300< LD50 ≤2000 mg/kg. The substance is classified in Category 4 for Acute Oral Toxicity, in accordance with EU CLP criteria.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Remarks:
This study was available at the time of registration and is submitted as part of an obligation to submit all relevant information on the substance. The study was not commisioned for REACH registration purposes.
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted by a GLP accredited laboratory using a test method eqiuivalent to OECD Testing Guideline 402.
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
not specified
GLP compliance:
yes
Test type:
standard acute method
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Acdemy of Militray Medical Sciences
- Weight at study initiation: 200 g
- Fasting period before study: 3 days
- Housing: SPF
- Acclimation period: 3 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20~25
- Humidity (%): 40%~70%

Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- % coverage: 40%
- Type of wrap if used: gauze

REMOVAL OF TEST SUBSTANCE
- Washing (if done): warm water
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2500mg/kg bw
- Constant volume or concentration used: no
- For solids, paste formed: no
Duration of exposure:
24 h
Doses:
2500 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
Duration of observation period following administration: 14 days
Frequency of observations and weighing: 0 d; 7 d; 14 d
Necropsy of survivors performed: no
Other examinations performed: clinical signs, body weight,organ weights, histopathology
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 500 mg/kg bw
Remarks on result:
other: No 95% CL information was provided
Mortality:
No deaths were observed.
Clinical signs:
No clinical sign were observed.
Body weight:
No effects were observed.

Summary of results

Animal

Dose Level

Animal

Weight (g, Average±SD)

Death

Mortality

LD50

Sex

(mg/kg bw)

Number

0 day

7 days

14 days

Number

%

(g/kg bw)

Male

2500

5

211.4 ± 5.7

253.2 ± 9.7

288.8 ± 9.8

0

0

>2500

Female

2500

5

213.2 ± 3.2

226.2 ± 4.4

237.2 ± 3.3

0

0

>2500

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
There were no deaths and no clinical signs were observed during the observation period. The LD50 found in the study was >2500 mg/kg bw, as such EMI-DBS is not classified as an acute dermal toxin in accordance with EU CLP criteria.
Executive summary:

The acute dermal toxicity of the test substance was determined using a method equivalent to the OECD Guideline for Testing of Chemicals 402. A dose of 2500 mg/kg bw was applied occlusively by gauze to the skin of male and female Wistar rats. The rats were exposed to the test substance for 24 hours. There were no deaths and no clinical signs were observed during the observation period. The LD50 found in the study was >2500 mg/kg bw, as such EMI-DBS is not classified as an acute dermal toxin in accordance with EU CLP criteria.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Additional information

A study was conducted on the registered substance to determine the acute oral toxicity of the substance. The acute oral toxicity of the test substance was determined in accordance with the OECD Guideline for Testing of Chemicals 423. The test substance was orally administered to female Sprague-Dawley rats. The first and second step doses were 300 mg/kg bw/day, the third step dose was 2000 mg/kg bw/day. The first step dose was chosen due to an absence of toxicity data on the test substance. Mortalities, clinical signs and body weight changes were monitored during the 14 -day observation period. Necropsy and gross findings were performed for all animals. There were no dead animals or unusal clinical signs or unusual observations at necropsy in the animals in the first and second steps (300 mg/kg). There were two dead animals 1 day after administration of the test substance in the third step (2000 mg/kg). Mucous stool and inanimation on the administration day were recorded as clinical signs in the third step. No abnormal gross findings were observed during necropsy. Based on the results of the study, EMI-DBS was found to have an LD50 of 300< LD50 ≤2000 mg/kg. The substance is classified in Category 4 for Acute Oral Toxicity, in accordance with EU CLP criteria.

A study was conducted on the registered substance to determine the acute dermal toxicity of the substance. The acute dermal toxicity of the test substance was determined using a method equivalent to the OECD Guideline for Testing of Chemicals 402. A dose of 2500 mg/kg bw was applied occlusively by gauze to the skin of male and female Wistar rats. The rats were exposed to the test substance for 24 hours. There were no deaths and no clinical signs were observed during the observation period. The LD50 found in the study was >2500 mg/kg bw, as such EMI-DBS is not classified as an acute dermal toxin in accordance with EU CLP criteria.

Justification for selection of acute toxicity – oral endpoint

This key study was performed in a GLP accredited laboratory using OECD Testing Guideline 423.

Justification for selection of acute toxicity – dermal endpoint

This key study was performed in a GLP accredited laboratory using a method equivalent to OECD Testing Guideline 402.

Justification for classification or non-classification

The test substance was found to have an LD50 of 300< LD50 ≤2000 mg/kg in the acute oral toxicity study. The substance is classified in Category 4 for Acute Oral Toxicity, in accordance with EU CLP criteria. The LD50 found in the acute dermal toxicity study was >2500 mg/kg bw, as such EMI-DBS is not classified as an acute dermal toxin in accordance with EU CLP criteria.

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