Ethylene bis[1,3-dihydro-1,3-dioxoisobenzofuran-5-carboxylate]

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was conducted between 7 May 2007 and 28 June 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

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

Test material information

Constituent 1

Reference substance name:

Ethylene bis[1,3-dihydro-1,3-dioxoisobenzofuran-5-carboxylate]

EC Number:

217-062-1

EC Name:

Ethylene bis[1,3-dihydro-1,3-dioxoisobenzofuran-5-carboxylate]

Cas Number:

1732-96-3

Molecular formula:

C20H10O10

IUPAC Name:

2-(1,3-dioxo-1,3-dihydro-2-benzofuran-5-carbonyloxy)ethyl 1,3-dioxo-1,3-dihydro-2-benzofuran-5-carboxylate

impurity 1

Reference substance name:

Benzene-1,2,4-tricarboxylic acid 1,2-anhydride

EC Number:

209-008-0

EC Name:

Benzene-1,2,4-tricarboxylic acid 1,2-anhydride

Cas Number:

552-30-7

Molecular formula:

C9H4O5

IUPAC Name:

Benzene-1,2,4-tricarboxylic acid 1,2-anhydride

impurity 2

Reference substance name:

not assignable

IUPAC Name:

not assignable

Specific details on test material used for the study:

Substance referred to as RIKACID TMEG-100
Chemical name is given as 1,2,4-Benzenetricarboxylic acid, ester with 1,2-ethanediol, but substance is actually Ethylene bis[1,3-dihydro-1,3-dioxoisobenzofuran-5-carboxylate]
Lot No. 0020
CAS No. 71342-70-6
Description: White-colored powder
Purity: 99.2%
Storage temperature: Room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

Number of animals (sex), age, body weight range (on receipt): 13 (females), 7-week old, 161.50 - 180.82g
Number of selected animals (sex), age, body weight range (at the start of administration): 6 (females), 8-week old, 179.11 - 198.52 g

Temporary identification for received animals
Upon receipt of animals, all animals were given a clinical inspection to health conditions. Body weights were measured using the automatic animal balance (CP3202S, Sartorious, Germany) and temporary identification numbers were marked on the tail using a red indelible pen.

Quarantine and acclimation
During the quarantine and acclimation period, animals were observed for clinical signs once a day. After they were acclimated to laboratory conditions for 7 days, health conditions were examined and released for study use by a staff veterinarian.

Group assignment
At the end of the quarantine and acclimation period, body weights were measured with the automatic animal balance (CP3202S, Sartorius, Germany) and each 3 healthy animals were assigned to respective steps in sequence of higher body weight.

Animal identification
The animals were identified individually by black indelible marking on the tail and each cage was compartmentalized by color-coded card enable to identify the step.

Housing conditions
Housing: Stainless wire cage (260W x 350D x 210H (mm))
Number of animals per cage: 1 (quarantine and acclimation period/observation period)
Temperature: 20.0 - 24.4 °C
Humidity(Relative humidity): 39.3 - 69.3%
Air exchanges: 10 - 15 air changes per hour (All fresh air method)
Lighting: The sequence being cycle of 12 hours light and 12 hours dark(light on : 7a.m., light off: 7p.m.)
Intensity of illumination: 150 - 300 Lux

Diet
Type: TEKLAD CERTIFIED GLOBAL 18 % PROTEIN RODENT DIET 2918C (Lot No. : 2918C 111406MA, 2918C 012207MA)
Supplier: Harlan TEKLAD, Madison, Wisconsin 53744-4220, USA
Method of supply: The diet was provided ad libitum.
The diet was analyzed periodically by manufacturer to quantify nutrient contents and contaminants. In addition, contaminant in the diet were identified twice a year in Korea Testing and Research Institute for Chemical Industry (539-3, Pyeongchon-dong, Daedeok-gu, Daejeon, Korea) to validate whether all analyzed items were within the contaminant range limit set in the test facility.

Drinking water
Type: Filtered and purified municipal tap water
Method of supply: Drinking water was provided ad libitum using a polycarbonate bottle (500 mL).
The quality of supplied water in Biotoxtech Co., Ltd. was analyzed twice a year in Chungcheongbuk-do Institute of Health and Environment Research (140-50 Songjeong-Dong, Cheongju-si, Chungcheongbuk-Do Korea) and biannual water reports was reviewed according to the Regulation of Quality Criteria for Potable Water and Test (Ministry of Environment Ordinance No. 122, 2002).

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

Corn oil was chosen as the vehicle since it was confirmed that the test substance was suspended into the vehicle through the preliminary solubility test to determine the solubility and dispersion characteristics of the test substance.

Preparation
On the day of administration, the test substance was weighed in semi-micro electronic balance (CP423S, Sartorious, Germany) and transferred to a mortar. A small amount of prepared vehicle was added and both materials were mixed. More vehicle material was added in an amount sufficient to bring the formulation to the desired volume. Formulations were prepared freshly each day.

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg body weigh

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: According to the MSDS for the test substance which LD50 in rats was above 5000 mg/kg, the dose of 2000 mg/kg body weight was selected as starting dose level for step 1.

Doses:

2000 mg/kg bodyweight

No. of animals per sex per dose:

Two groups of 3 animals were treated with 2000 mg/kg bodyweight

Control animals:

no

Details on study design:

Administration method
Rats were overnight fasted for approximately 16 hours prior to dosing, but drinking water was not withheld. Following the period of fasting, individual dose was calculated for each animal based on the animal's fasted body weight taken just before test substance administration and orally administered once using a sonde attached to a disposable syringe (1 mL ). The diet was provided 4 hours after the completion of the administration.

Dose
The dose of 2000 mg/kg body weight was selected as starting dose level for step 1. Based on the results for mortality at the previous dose level (no dead found during 3 days observation after treatment), 2000 mg/kg dose was administered for a following step (step 2) in accordance with an 'TCCA-Good Laboratory Practice Standards and Test Guidelines, Annex 1 (TCCA(Toxic Chemical Control Act)-Good Laboratory Practice) [NIER Public Notice No. 2006-29] (Korean National Institute of Environmental Research, December 19, 2006 )
A constant dosage volume of 5 mL/kg body weight was administered.


Observation and pathology
The commencing day of administration was designated as Day 1.

Clinical signs
The animals were observed for 30 minutes continuously after dosing, at 1, 2, 4 and 6 hours, and thereafter once daily for 14 days (from Day 2 to Day 15) after dosing.

Body weights
Body weights were measured prior to dosing (Day 1), at Day 4, 8 and the day of - necropsy (Day 15) using the automatic animal balance (CP3202S, Sartorius, Germany).

Necropsy
After the end of the observation period, all animals were exsanguinated by transecting the abdominal aorta under ether anesthesia and observed macroscopically about full external surface, cranial cavity and organs of thoracic and abdominal cavities.

Statistics:

Other than mean value calculations from the data obtained through this study, no statistical analysis was not performed.

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 in all animals for step 1 and 2.

Clinical signs:

other: In all animals treated with 2000 mg/kg for step 1, no clinical abnormalities were observed throughout the study. Soft stool was observed on Day 2 in all animals treated with 2000 mg/kg for step 2. No other clinical abnormalities were observed through the

Gross pathology:

In the necropsy for step 1 and 2, there were no macroscopic findings on external surface and organ of body cavity (cranial, thoracic and abdominal).

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, the test substance, RIKACID TMEG-100, was classified as 'Category 5 or Unclassified' by the 'TCCA-Good Laboratory Practice Standards and Test Guidelines, Annex 1 (TCCA(Toxic Chemical Control Act)-Good Laboratory Practice)'.
LD50 > 2000 mg/kg

Executive summary:

The study was conducted to evaluate the toxic effect and LD50 value of the test substance, RIKACID TMEG-100, following single oral administration to female rats.

Each three healthy female Sprague-Dawley rats were assigned to the test group for step 1 and 2. Three female rats were overnight fasted and administered orally at the starting dose level of 2000 mg/kg for step 1. Based on the mortality from step 1 (no found dead), 2000 mg/kg dose was administered successively to three female rats for step 2.

The animals were monitored for clinical signs and body weight changes during 14 days observation period after administration and were subjected to gross necropsy on Day 15.

During the observation period, there was not observed any mortality from all animals treated with 2000 mg/kg of the test substance for step 1 and 2.

No clinical abnormalities were observed in animals treated with 2000 mg/kg of the test substance for step 1. Soft stool were observed on Day 2 from rats treated with 2000 mg/kg for step 2, but no other clinical abnormalities were observed through the remainder of study. Any abnormalities in body weight changes were not observed in all animals treated with 2000 mg/kg of the test substance.

The gross necropsy examinations in all animals treated with 2000mg/kg did not reveal any macroscopic findings attributed to the treatment with the test substance.

In conclusion, the test substance, RIKACID TMEG-100, was classified as 'Category 5 or Unclassified' by the 'TCCA-Good Laboratory Practice Standards and Test Guidelines, Annex 1 (TCCA(Toxic Chemical Control Act)-Good Laboratory Practice)'.