Indoline-2,3-dione

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 March 2015 - 31 March 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test method according to OECD Guideline 420. GLP study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Indoline-2,3-dione

EC Number:

202-077-8

EC Name:

Indoline-2,3-dione

Cas Number:

91-56-5

Molecular formula:

C8H5NO2

IUPAC Name:

2,3-dihydro-1H-indole-2,3-dione

Details on test material:

- Name of test material (as cited in study report): Indoline-2,3-dione
- Physical state: orange, crystalline powder
- Analytical purity: 98.0%
- Lot/batch No.: MP 1076.3
- Storage condition of test material: storage temperature: 5-35ºC.

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Experimental Medicine Centre at the Medical University in Białystok
- Age at study initiation: 11 weeks old
- Weight at study initiation: 207.3 g
- Fasting period before study:
- Housing: plastic cages covered with wi re bar lids. The dimensions of the cages were 58 x 37 x 21 cm (length x width x height).
- Diet (e.g. ad libitum): “Murigran” standard granulated laboratory fodder, ad libitum.
- Water (e.g. ad libitum): drinking tap water, ad libitum.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 – 23 °C
- Humidity (%): 40 – 58%
- Air changes (per hr): about 16 times/hour
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 400 mg/mL
- Amount of vehicle (if gavage): 1 mL

MAXIMUM DOSE VOLUME APPLIED: 0.5 mL/100 g b.w.

- Rationale for the selection of the starting dose: The starting dose for the preliminary experiment was selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg b.w. as a dose expected to produce evident toxicity. Since no data were available, the preliminary experiment started with the administration of the test item at a dose of 300 mg/kg b.w to one female rat. Since no evident toxicity was produced, the test item at a single dose of 2000 mg/kg b.w. was administered to the second animal. The animal treated with the test item at a dose of 2000 mg/kg b.w. survived the experiment. On the grounds of the preliminary experiment results, the dose of 2000 mg/kg b.w. was selected to be used in the main experiment.

Doses:

Sighting study: 300 and 2000 mg/kg b.w.
Main study: 2000 mg/kg b.w.

No. of animals per sex per dose:

2 females in the sighting study.
4+1 (from the preliminary study) females in the main study.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: body weights were determined on days 0 (directly before the administration of the test item), 7, and 14 (before euthanasia).
- Necropsy of survivors performed: yes.
- Other examinations performed:
General condition: the observation of all animals for morbidity and mortality was conducted twice a day or once a day (on days off) during the 14-day experiment.
Detailed clinical observations: the detailed clinical observations were performed on the day of the test item administration (day 0), i.e. 10, 30, and 60 minutes after the administration and then at hourly intervals up to the 5th hour after the administration. From the 1st to the 14th day of the experiment, the detailed clinical observations were performed once a day.
Gross examinations: After the 14-day observation period, all animals were euthanized by intraperitoneal administration of Morbital at a dose of 200 mg/kg b.w. and subjected to gross examinations. The detailed gross examinations involved the observation of the external body surface, all natural apertures, and the cranial, thoracic, and abdominal cavities with their content.

Results and discussion

Preliminary study:

Following single administration of the test item at a dose of 300 mg/kg b.w. to the first animal used in the preliminary experiment, no signs of toxicity were stated. The animal survived the experiment.
Following single administration of the test item at a dose of 2000 mg/kg b.w. to the other animal used in the preliminary experiment, the rounded back, slightly decreased locomotor activity, and dejection were stated on the administration day. The animal survived the experiment.

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All animals survived the experiment.

Clinical signs:

other: The rounded back, slightly decreased locomotor activity, and dejection were stated in all animals on the administration day.

Gross pathology:

The animals did not exhibit any pathological changes.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

No mortality was observed following single administration of the test item at a dose of 2000 mg/kg bw in rats. The test substance is not classified according to the CLP Regulation (EC) no 1272/2008.

Executive summary:

The acute oral toxicity study on Wistar rats was performed according to the OECD Guideline 420 and EU Method B.1. bis, following the Principles of GLP. The test item was prepared as an oil suspension in a volume of 0.5 mL/100 b.w. and it was administered with a metal stomach tube. A preliminary experiment started with the administration of 300 mg/kg b.w. of the test item to one animal. Since no evident toxicity was observed, a second animal was dosed at 2000 mg/kg b.w. The animal survived the experiment but presented signs of toxicity: rounded back, slightly decreased locomotor activity, and dejection were stated on the administration day.

In the main experiment 4 animals received 2000 mg/kg b.w of the test item. After the administration of the test item, the animals were observed for 14 days. General and detailed clinical observations were conducted daily during the entire experiment. Body weights of the animals were determined on days 0, 7, and 14. All the animals survived the experiment and were euthanized after the observation period. Gross examinations did not reveal any pathological changes in the examined animals. On the basis of these results, the test substance is not classified according to the CLP Regulation (EC) no 1272/2008.