Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22.09. – 09.10.2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid
Details on test material:
Expiration: 03/2029
Storage condition of test material: at the room temperature in tightly closed container

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Wistar CRL, monitored quality
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: SPF breeding, VELAZ s.r.o., Únětice, Czech Republic, RČH CZ 11760500
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: 217 - 224 g
- Housing: animal room with monitored conditions – 3 animals of one sex in one plastic breeding cage with sterilized shavings of soft wood
- Diet (e.g. ad libitum): pelleted standard diet for experimental animals ad libitum
- Water (e.g. ad libitum): drinking tap water ad libitum (quality corresponding to Regulation No. 252/2004 Czech Coll. of Law)
- Acclimation period: 13 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3°C, permanently monitored
- Humidity: 30 – 70 %, permanently monitored
- Photoperiod: 12 hour light/12 hour dark

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
Olivae oleum raffinatum - Lot/batch no.: 5523502 (expiration 09/2015), producer Dr. Kulich Pharma, s.r.o. Hradec Králové, Czech Republic

DOSING:
Test procedure with a starting dose of 300 mg/kg was selected. This level was chosen as a starting dose according the test guideline. The test substance in this dose level was administered sequentially to one group of 3 females. No death of animals was observed therefore the testing was continued with dose 2000 mg/kg (application with time distance 24 hours). Because no death of animals was observed, next step using the same dose 2000 mg/kg was performed as a confirmation.

Testing schedule: START: 300 mg/kg – 3 females (Step No.1): no deaths ► 2000 mg/kg – 3 females (Step No. 2): no deaths ► 2000 mg/kg – 3 females (Step No. 3): no deaths ► END of study

PREPARATION AND APPLICATION OF THE TEST SUBSTANCE
Immediately before application the test substance was weighed and mixed in vehicle. The test substance was administered suspended in olive oil (pharmaceutical quality).
The test substance was administered to the stomach by tube. The single volume of administered suspension was 1 mL/100 g of animal body weight. The starting dose was 300 mg/kg of body weight.
Doses:
300 mg/kg (1st step)
2000 mg/kg (2nd step)
2000 mg/kg (3rd step)
No. of animals per sex per dose:
3 animals per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 1st day: twice (30 minutes and 3 hours after application), 2nd day: twice (in the morning and in the afternoon), thereafter days: once a day
Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.

- Necropsy of survivors performed: yes
- examinations performed:
nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated. All gross macroscopic changes of organs and tissues were recorded on special data sheets.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No death of animals.
Clinical signs:
No clinical signs of intoxication were observed during the study.
Body weight:
Body weight was recorded and weight increments were calculated in all animals.
Weight increments were adequate to species, sex and age of animals in experiment. Only in female No. 4 reduction of body weight between 8th and 15th day was observed.
Gross pathology:
No pathologic macroscopic changes were diagnosed during pathological examination.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance toxicity was evaluated on the basis of mortality, clinical signs of intoxication, body weight increments during the observation period and necropsy findings at the end of study.
The test substance administered at the dose of 2000 mg/kg caused no death of animals.
No clinical signs of intoxication were observed during the study No pathologic macroscopic changes were diagnosed during pathological examination.
According to the study results the value of LD50 of the test substance, 2-(3´,5´-Dichloro-2´-hydroxyphenyl)-4-quinazolinone, for female rats is higher than 2000 mg/kg of body weight.
Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance, 2-(3´,5´-Dichloro-2´-hydroxyphenyl)-4-quinazolinone after a single oral administration to Wistar CRL rats.

The testing was performed according Method B.1 tris: Acute Oral Toxicity - Acute Toxic Class Method, Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

The test substance was administered suspended in olive oil (pharmaceutical quality) as a single dose, given orally via gavage to three groups of three female Wistar CRL rats.

The dosing was performed sequentially in three groups of three females: group No. 1 - first step using the starting dose of 300 mg/kg of body weight, group No. 2 - second step using higher dose 2000 mg/kg and group No. 3 – third step using the same dose 2000 mg/kg.

The test substance administered at the dose of 2000 mg/kg caused no death of animals. No clinical signs of intoxication were detected during whole study. No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results the value of LD50 of the test substance, 2-(3´,5´-Dichloro-2´-hydroxyphenyl)-4-quinazolinone, for female rats is higher than 2000 mg/kg of body weight.