Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From September 04, 2017 to September 17, 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 guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
other: Guidance Document on Acute Oral Toxicity Testing, OECD Series on Testing and Assessment No 24, 2001
Deviations:
no
Qualifier:
according to
Guideline:
other: Guidance Document on the Recognition, Assessment and Use of Clinical Signs as Humane Endpoints for Experimental Animals Used in Safety Evaluation. Environmental Health and Safety Publications Series on Testing and Assessment No 19, 2000
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
Lot No.: WD0066442; Purity: 100 % (UVCB); Appearance: homogeneous, amber liquid

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Species: Wistar rats
Source: Dobrá Voda, Slovak Republic
Age at First Dose: 8-12 weeks; female animals were non-pregnant and nulliparous
Animal Health: health condition of animals was examined by a veterinarian before initiation of the study.
Acclimation: The animals were acclimated under the conditions identical to the conditions during the experiment 5 days prior to the start of treatment. The acclimation was according to the standard operation procedure.
Housing Condition: The animals were housed in plastic cages suspended on stainless steel racks, up to 3 animals per cage in a room equipped with central air conditioning. The average room temperature was maintained within the range of 23.2 ± 0.3 °C, relative humidity within 55.3 ± 2.5 %. The light regimen was set to a 12-hour light /12-hour dark cycle. Sanitation was performed according to the standard operation procedures.
Diet: The laboratory food ssniff (Spezialdiäten GmbH, Germany) was offered at recommended doses each day approximately at the same time. The certificate of analysis is included in the raw data.
Water: The animals received tap water for human consumption. Supply of drinking was unlimited. The quality of drinking water is periodical analysed and recorded; certificate of analysis is included in the raw data.
Bedding: Lignocel S3/4, Lufa - ITL GmbH, Germany.
Animals Identification: The animals in the cage were marked by a line (I-III) on the tail with a waterproof marker. Each cage was marked with the study code, ID of animals and date of administration of the test substance.
Justification forthe Choice of Species: Normally females are used for testing OECD TG 423 because females are typically the more sensitive gender.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
Dose Preparation: The required amount of the test substance (according to the body weight and dose) was mixed with vehicle (olive oil) shortly before administration.
Dose Administration: The test substance was administered in a single dose by gavage using a metal stomach tube. Animals were fasted prior to dosing (food but not water were withheld overnight). Following a period of fasting, animals were weighed and the test substance administered. After test article administration, food was withheld for further 3-4 hours.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 females
Control animals:
no
Details on study design:
A limit dose of 2000 mg/kg bw was used as a starting dose. One group of 3 females was dosed. Test item-related mortality was not observed during 24 hours and therefore in a second step another 3 females were treated at the same dose.

Clinical Observation: Animals were observed individually immediately after administration of the test substance and 0.5, 1, 2, and 4 hours later. Each animal was inspected daily for the next 14 days.
Observations included: changes in skin, fur, eyes, mucous membranes, respiratory, circulatory, autonomic and central nervous systems, somatomotor activity, and behavioural pattern. Particular attention was given to potential neurologic endpoints such as tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Body Weight: Individual weights of animals were measured immediately prior to test substance was administered and weekly thereafter. Weight differences after first and second weeks after administration were calculated and recorded.
Necropsy: All test animals were subjected to gross necropsy and the results were recorded for each animal.

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 mortality was observed during the study.
Clinical signs:
During the follow up period, no animals displayed signs of intoxication, change of health, nor any other adverse reactions.
Body weight:
The body weights of 5/6 animals slightly increased during the study. Stagnation of body weight in three animals between the first and second week was observed.
Gross pathology:
All animals were necropsied. During necropsy, no macroscopic findings were observed.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the study conditions, the LD50 of the test substance was > 2000 mg/kg bw after single oral administration to Wistar rats.
Executive summary:

A study was conducted to determine the potential toxic effect of the test substance when administered as a single oral dose to Wistar rats according to the OECD Guideline 423 Acute Toxic Class (ATC) method, in compliance with GLP. Available information indicated that the test substance is likely to be non-toxic, therefore, 2000 mg/kg bw was used as a starting dose. Two groups of 3 females received the test substance and all 6 survived. The limit dose of 2000 mg/kg bw did not cause death, evident signs of toxicity during the 14 d observation period. Stagnation of bodyweight in 3 animals between the first and second week was observed. During necropsy, no macroscopic findings were noted. Under the study conditions, the LD50 of the test substance was > 2000 mg/kg bw after single oral administration to Wistar rats (Hozova, 2017).