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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06.11.2017 – 23.11.2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Tetrazol-5-ylamine
EC Number:
224-581-7
EC Name:
Tetrazol-5-ylamine
Cas Number:
4418-61-5
Molecular formula:
CH3N5
IUPAC Name:
1H-tetrazol-5-amine
impurity 1
Reference substance name:
Unknown impurities
Molecular formula:
Not available
IUPAC Name:
Unknown impurities
Test material form:
solid: crystalline
Details on test material:
white crystals

Test animals

Species:
rat
Strain:
Wistar
Remarks:
Wistar Han, monitored quality
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: breeding farm VELAZ s.r.o., Lysolajské údolí 15/53, Czech Republic, RČH CZ 11760500
- Age at study initiation: 8 weeks
- Weight at study initiation: 178 - 190 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 water ad libitum (quality corresponding to Regulation No. 252/2004 Czech Coll. of Law)
- Acclimation period: 12 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
Aqua pro iniectione - Lot/batch no.: 1703240253 (expiration 03/2019), producer Ardeapharma Ševětín, Czech Republic

DOSING:
The dose level of 300 mg/kg of body weight was used as the starting dose, according to the test guideline, because there was no information about toxicity from sponsor. This dose caused no death of 3 females in group No.1. The dose of 2000 mg/kg level was sequentially applied (application with time distance 24 hours) to 3 females in group No. 2. Because this dose caused no death of females, the same dose of 2000 mg/kg level was sequentially applied for confirmation to 3 females in group No. 3. No death of animals was observed in this group.

PREPARATION AND APPLICATION OF THE TEST SUBSTANCE
Immediately before application the test substance was weighed, mixed in vehicle (aqua pro iniectione) and resulting suspension was administered to the stomach by tube.
Doses:
300 mg/kg (1st step)
2000 mg/kg (2nd step)
2000 mg/kg (confirmation)
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
- Other 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 in all 9 females during the clinical observation after the application.
Body weight:
Body weight was recorded and weight increments were calculated in all surviving animals at the end of study. Weight increments were adequate to species, sex and age of animals in experiment. No reduction of body weight 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 item 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 item administered at the dose of 300 mg/kg and 2000mg/kg caused no death of animals. No serious clinical signs of intoxication were detected at these doses during whole study. Weight increments were adequate to species, sex and age of animals in experiment.
No pathologic macroscopic changes were diagnosed during pathological examination.
According to the study results the value of LD50 of the test item, 5-aminotetrazole, 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 item 5-aminotetrazole, after a single oral administration to Wistar Han 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 item was administered in a single dose as a suspension in vehicle (Aqua pro iniectione), given orally via gavage to female Wistar rats. The volume of administered suspension was 1 ml/100 g body weight of animals.

The dosing was performed sequentially in three groups of three females: Group No. 1- the first step - using the starting dose of 300 mg/kg of body weight, this dose caused no death of animals. Group No. 2 - the second step using higher dose of 2000 mg/kg, this dose caused no death of animals, therefore the dose of 2000 mg/kg was sequentially applied for confirmation to group No. 3 in the third step.

The test item administered at the dose of 2000 mg/kg of body weight caused no death of all 6 females in group No. 2 a No. 3.

No clinical signs of intoxication were detected during the whole study in all 9 animals.

No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results, the value of LD50 of the test item, 5-aminotetrazole, for female rats is higher than 2000 mg/kg of body weight.

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