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Diss Factsheets

Administrative data

acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 May 2019 to 14 June 2019
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Relative humidity values and temperatures recorded slightly exceeded the expected range. The staff veterinarian confirmed that this would have no effect on the animals nor the study.
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:

Test material

Constituent 1
Chemical structure
Reference substance name:
3-tert-butyl-1-(3,5-dicarboxyphenyl)-1H-pyrazol-5-aminium chloride
EC Number:
Molecular formula:
3-tert-butyl-1-(3,5-dicarboxyphenyl)-1H-pyrazol-5-aminium chloride
Test material form:
solid: particulate/powder

Test animals

Details on test animals or test system and environmental conditions:
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sandhofer Weg 7, D-97633 Sulzfeld
- Females nulliparous and non-pregnant: yes
- Age at study initiation: ~10 weeks
- Weight at study initiation: 226 - 251 g
- Fasting period before study: yes. not fed the night before treatment
- Housing: Type II polypropylene/polycarbonate with bedding (SAFE 3/4 S certified wooden chips (J. Rettenmaier & Söhne GmbH + CO. KG, 73494 Holzmühle 1, Rosenberg, Germany)) available to animals during the study. The quality of the bedding was guaranteed by the supplier.
- Diet : Animals received ssniff SM R/M "Autoclavable complete diet for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494 Soest, Germany, ad libitum, except for the night before treatment. The food is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.
- Water : Animals received tap water from the municipal supply from 500 mL bottles, ad libitum. The water was fit for human consumption and was considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study. Water quality control analysis is performed once every three months and microbiological assessment is performed monthly, by Veszprém County Institute of State Public Health and Medical Officer Service (ÁNTSZ, H-8200 Veszprém, József Attila u. 36, Hungary).
- Acclimation period: at least 19 days

- Temperature (°C): 18.89 - 24.7ºC
- Humidity (%): 34 - 84 %
- Air changes (per hr): 15 - 20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12hrs dark/ 12hrs light per day.

Administration / exposure

Route of administration:
oral: gavage
Details on oral exposure:
- Concentration in vehicle: constant dose volume of 20ml/kg
- Justification for choice of vehicle: The solubility of the test item was examined during a short trial formulation. In order of preference, recommended vehicles were tested as: distilled water and 1% methyl cellulose. The formulation with distilled water was inhomogeneous. Formulation with 1% methylcellulose (abbreviated as 1% MC) was appropriate for treatment; therefore, it was found to be a suitable vehicle for formulations.

- Rationale for the selection of the starting dose:
Initially three females (Group 1) were treated at a dose level of 300 mg/kg bw. Approximately after 24 hours another 3 animals were treated at the same dose level. No mortality was observed therefore the third group of animals (Group 3) was treated at a dose level of 2000 mg/kg bw. Approximately after 24 hours another 3 animals were treated at the same dose level. No mortality was observed in the confirmatory group, therefore no further testing was required according to the test guidelines (OECD 423, Commission Regulation (EC) No 440/2008 of 30 May 2008, B.1.tris).
300 mg/kg bw 2000 mg/kg bw
No. of animals per sex per dose:
3 females 0 males
Control animals:
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were performed on all animals at 30 minutes, 1, 2, 3, 4 and 6 hours after dosing and daily for 14 days thereafter. The body weights were recorded on Days -1 (prior to removal of food), 0 (prior to administration), 7 and 14 with a precision of 1 g. Terminal body weight of animals were recorded prior to euthanasia.
- Necropsy of survivors performed: yes
- Other examinations performed: Clinical signs, body weight, body weight gain and gross macroscopic

Results and discussion

Effect levels
Key result
Dose descriptor:
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
The test item did not cause mortality at a dose levels of 300 and 2000 mg/kg bw.
Clinical signs:
After treatment at the dose levels of 300 and 2000 mg/kg bw, all the animals were symptom-free (including the observation period).
Body weight:
There were no effects on body weight or body weight gains in the animals.
Gross pathology:
There was no evidence of any macroscopic observations in the terminal animals at dose level of 300 nor 2000 mg/kg bw.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Under the conditions of this study, the acute oral LD50 value of the test item was found to be greater than 2000 mg/kg bw in female Crl:WI rats.

According to the GHS criteria, the test item can be ranked as not classified for acute oral exposure.