Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
14/02/2018 - 07/03/2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Chemrign / batch 02/16
- Expiration date of the lot/batch: 01/01/2021
- Purity test date: 100%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark
- Stability under test conditions: The test item was formulated within 2 hours ogf being applied to the test system. It is assumed that the formulation was stable for this duration.
- Solubility and stability of the test substance in the solvent/vehicle: The test item was freshly prepared as a solution in arachis oil BP. Arachis oil BP was used beacause the test item did not dissolve/suspend in distilled water.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium:

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was freshly prepared as a solution in arachis oil BP.
- Preliminary purification step (if any):
- Final dilution of a dissolved solid, stock liquid or gel:
- Final preparation of a solid:

FORM AS APPLIED IN THE TEST (if different from that of starting material)

TYPE OF BIOCIDE/PESTICIDE FORMULATION (if applicable)

OTHER SPECIFICS:

Test animals

Species:
rat
Strain:
Wistar
Remarks:
RccHan : WIST from Envigo RMS(US)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks
- Weight at study initiation: The body weight variataion did not exceed +/- 20% of the mean body weight of any previously treated animals
- Fasting period before study: an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing
- Housing: suspended solid-floor polypropylene cages furnished with wood flakes
- Diet (e.g. ad libitum) and Water (e.g. ad libitum): yes with the exception of an overnight fast immediately before dosing and for approximately 3 to 4 hours after dosing
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70% relative humidity
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE : Arachis oil

- Justification for choice of vehicle: Arachis oil BP was used beacause the test item did not dissolve/suspend in distilled water.

All animals were dosed once only by gavade, using a metal canula attached to graduated syringe.

Rationale for the selection of the starting dose :
In the absence of data regarding the toxicity of the test item, 300 mg/kg was chosen as the starting dose.
Doses:
300 mg/kg, 2000 mg/kg
No. of animals per sex per dose:
1 female rat for the Dose Level 2000 mg/kg
5 females rat for the Dose Level 300 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of clinical observations: 30 minutes, 1, 2 and 4 hours after dosing and then daily for up to 14 Days.
- Frequency of mobidity and mortality : twice daily, early and late during normal working days, and once daily at weekends and public holidays.
- Frequency of body weights : recorded on Day 0 and on Day 7 and 14 or at death.
- Necropsy of survivors performed: yes, gross necropsy : external examination and opening of the abbominal and thoracic cavities. No tissues were retained. Only the appareance of any macroscopic abnormalities was recorded.
- Euthanasia : cervical dislocation
Statistics:
NA

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - <= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Dose level - 300mg/kg
There were no deaths.
Dose level - 2000mg/kg
The animal was killed for human reasons, due to the occurence of clinical signs of toxicity that approached the severity limit set forth in the UK Home Office Project License.
Clinical signs:
Dose level - 300mg/kg
Signs of systemic toxicity noted in four animals during the day of dosing were hunched posture, ataxia and pilo-erection.
One animal appeared normal throughout the observation period.
Dose level - 2000mg/kg
Signs of systemic toxicity noted were hunched posture, lethargy, ataxia, occasional body tremos, prostration, decreasded respiratory rate, hyperthermia and ptosis.
Body weight:
Dose level - 300mg/kg
All animals showed expected gains in body weight over the observation period.

Gross pathology:
Dose level - 300mg/kg
No abnormalities were noted at necropsy
Dose level - 2000mg/kg
Abnormalities noted at necropsy were dark liver, dark kidney and a pale stomach.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The LD50 of the test item in the female Wistar strain rat was estimated to be in the range of 300-2000 mg/kg bw.
Executive summary:

An acute oral toxicity in the Wistar strain female rat (A. Sanders, 2018, Envigo) has been performed, according the 420 OECD guideline and in compliance with GLP, in order to assess the LD50 of the test item.

Following a sighting test with 1 animal at dose levels of 300 and 2000 mg/kg, a further group of 4 fasted female was given a single oral dose by gavage of test item, as a dissolution in arachis oil BP, at a dose level of 300 mg/kg. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

The animal treated at the dose level of 2000 mg/kg was killed for human reasons due to the occurenc of clinical signs of toxicity.

At the dose level of 300 mg/kg, there were no deaths, all animals showed expected gains in body weight and no abnormalities were noted at necropsy. Animals showed signs of systemic toxicity as hunched posture, ataxia and pilo-erection. One animal appeared normal throughout the observation period.

Therefore the LD50 ot the test item in the female Wistar strain rat is estimated to be in the range of 300-2000 mg/kg bw.

The test item is classified as Category 4 according to GHS criteria.