Ethyl 1,3-dihydro-1,3-dioxo-2H-isoindole-2-carboxylate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 March 2016 - 05 April 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

2001

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Version / remarks:

#440/2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1100 (Acute Oral Toxicity)

Version / remarks:

2002

Deviations:

no

Test guideline 4

Qualifier:

according to guideline

Guideline:

other: JMAFF

Version / remarks:

2000, including partial revisions

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl 1,3-dihydro-1,3-dioxo-2H-isoindole-2-carboxylate

EC Number:

245-048-5

EC Name:

Ethyl 1,3-dihydro-1,3-dioxo-2H-isoindole-2-carboxylate

Cas Number:

22509-74-6

Molecular formula:

C11H9NO4

IUPAC Name:

ethyl 1,3-dioxo-2,3-dihydro-1H-isoindole-2-carboxylate

Test material form:

solid

Details on test material:

- Appearance: white solid
- Storage conditions of test material: in refrigerator (2-8°C)

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Stability under test conditions: stable
- Solubility and stability of the test substance in the solvent/vehicle: soluble in propylene glycol

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The preparations (w/w) were kept at room temperature and were dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies. Adjustment was made for specific gravity of the vehicle (1.036). No correction was made for purity of the test item.

OTHER SPECIFICS:
pH (1% in water, indicative range)
4.36 – 3.38 (determined by WIL Research Europe)

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: ca. 8 weeks
- Weight at study initiation: 143-153 g
- Fasting period before study: yes, overnight until 3-4 hours after the administration
- Housing: group housing of 3 animals per cage in labeled Makrolon cages (MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom).
- Diet (e.g. ad libitum): pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.7-21.4
- Humidity (%): 42-50
- Air changes (per hr): at leat 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 16 March 2016 To: 05 April 2016

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: The vehicle was selected based on trial preparations performed at WIL Research Europe and on test item data supplied by the Sponsor.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION (if unusual): The preparations (w/w) were kept at room temperature and were dosed within 4 hours after adding the vehicle to the test item. Homogeneity was assessed by visual inspection of the solutions and the formulations were stirred during dosing, which ensures homogeneity sufficient for these kinds of studies. Adjustment was made for specific gravity of the vehicle (1.036). No correction was made for purity of the test item.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: based on the cut-off value set by OECD Guideline 423

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3 females/dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Twice daily for mortality. Clinical signs were checked at periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15. Body weights were determined on Days 1 (pre-administration), 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weights

Statistics:

Not performed (the method is not intended to establish LD50)

Results and discussion

Preliminary study:

Not performed

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

other: Hunched posture and piloerection were noted in the three animals on Day 1.

Gross pathology:

No abnormalities were found at macroscopic post mortem examination of the animals.

Applicant's summary and conclusion

Interpretation of results:

other: not classified under Regulation 1272/2008/EC

Conclusions:

In a GLP-compliant guildeline study, acute oral LD50 of the test substance was above 2000 mg/kg bw.

Executive summary:

In a GLP-compliant OECD guideline study 423, the test substance was administered at the limit dose of 2000 mg/kg bw to a group of three female rats. There were no mortalities. Hunched posture and piloerection were noted the three animals on Day 1. The body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain. No abnormalities were found at macroscopic post mortem examination of the animals. Based on the results of the study, the oral LD50 was established to exceed 2000 mg/kg bw. No classification of the substance for acute toxicity is warranted based on these results according to Regulation 1272/2008/EC.