Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
22.3.2010 - 5.5.2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was carried out in accordance with internationally valid GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Reaction product of Distillates (petroleum), acid-treated heavy naphthenic and calcium oxide
- Molecular formula: not known - UVCB substance
- Molecular weight: not known - UVCB substance
- Smiles notation: not known - UVCB substance
- InChl: not known - UVCB substance
- Substance type: technical product
- Physical state: solid
- Lot/batch No.: 26.2.2010
- Expiration date of the lot/batch: 26.2.2012
- Stability under test conditions: stable
- Storage condition of test material: The substance was stored in PE bottle at room temperature.


Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:breeding farm VELAZ s.r.o., Koleč u Kladna, Czech Republic, RČH CZ 21760152
- Age at study initiation: 8 -10 weeks at the time application
- Weight at study initiation: 144 - 158 g
- Fasting period before study: 12 hours
- Housing: animal room with monitoring conditions – 3 animals of one sex in one plastic breeding cage Velaz T4
- Diet (e.g. ad libitum): ST 1 BERGMAN – standard pelleted diet 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: a least 5 days
- Bedding: sterilized shavings of soft wood
- Randomisation: according to the internal rule, at the start of the study the weight variation of animals was minimal and did not exceed ± 20 % of the mean weight
- Identification of animals: colour marks 1 - 3 on tail of animals, each cage was marked with the number of study and dose of the test substance
- Health condition: certificate of good health condition – from breeding farm; no signs of diseases were observed at clinical check-in, during the acclimatisation period and before the start of study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): room temperature 22 +/- 3°C, permanently monitored
- Humidity (%): relative humidity 30 – 70 %, permanently monitored
- Photoperiod (12 hours light/12 hours dark)

TIME SCHEDULE OF OBSERVATIONS:
Body weight: before application, the 8th day and before euthanasia of animals
Mortality: daily
Clinical examination: daily
Pathological examination: 15th day

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE
Preparation and application of the test substance: Immediately before application the test substance was weighed, mixed in vehicle (olive oil) and resulting suspension was administered to the stomach by tube. The single volume of administered suspension was 1mL/100 g of animal body weight.

- Justification for choice of vehicle: applicable suspension
Olive oil – Oleum olivae
Batch No.: 4726901
Expiration: 08/2011
Producer: Dr. Kulich, Pharma Hradec Králové, Czech Republic

MAXIMUM DOSE VOLUME APPLIED: 1mL/100 g of animal body weight

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Test procedure with a starting dose of 2000 mg/kg was selected.
Testing schedule (according to EU Method B.1 tris Annex 1D)
START: 2000 mg/kg – 3 females (Step No.1): no deaths ► 2000 mg/kg – 3 females (Step No. 2): no deaths ► END of study
Doses:
2000 mg/kg of animal body weight
No. of animals per sex per dose:
6 animals - females
Step No. 1: 3 females
Step No. 2: 3 females

Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of weighing: Animals were weighed before application, on the 8th day of study and at day 15, before euthanasia of animals.
- Frequency of observation: After application the animals were observed individually – the first day: twice (30 minutes and 3 hours after application), the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days.
Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.
- Necropsy of survivors performed: yes
All test animals survived to the end of study were sacrificed on the 15th day by prolonged ethereal narcosis and gross necropsy was carried out. Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated.


Results and discussion

Preliminary study:
no
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality
Clinical signs:
No clinical signs of intoxication.
Body weight:
Weight increments were adequate to species, sex and age of animals in experiment.
Gross pathology:
The test substance caused no pathological changes in all animals from both groups.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance - Reaction product of Distillates (petroleum), acid-treated heavy naphthenic and calcium oxide - after a single oral administration to Wistar rats.

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

According to the study results the value of LD50 of the test substance for female rats is higher than 2000 mg/kg of body weight.

Test substance was administered in a single dose as a solution in olive oil (vehicle), given orally via gavage to two groups of three female Wistar rats. The dosing was performed sequentially in two groups of three females: group No.1 - first step using the starting dose of 2000 mg/kg of body weight and group No.2 - second step using the same dose.

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