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Administrative data

Description of key information

Acute oral toxicity: LD50 (female) >2000 mg/kg bw (OECD 423/GLP)


Acute dermal toxicity: LD50 (female) >2000 mg/kg bw (OECD 402/GLP)

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 May 2020 - 11 September 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: test material provided by sponsor: 91112Y
- Expiration date of the lot/batch: 12 March 2021
- Purity test date: 92.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25℃, ≤70% relative humidity)
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: 1% methylcellulose was appropriate for treatment

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was freshly formulated at concentrations of 30 and 200 mg/mL, stirred with a magnetic stirrer during an overnight period before the treatment.
Species:
rat
Strain:
other: Crl:WI (Wistar)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH (Address: Sandhofer Weg 7, D-97633 Sulzfeld, Germany)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young healthy adult rats, 8-11 weeks old
- Weight at study initiation: 196 - 246 g
- Fasting period before study: At least 5 days
- Housing: Type II. or III. polycarbonate cages (3 animals/cage), Certified laboratory wood bedding and nest building material.
- Diet: ssniff SM R/M "Autoclavable complete diet ad libitum
- Water: tap water from the municipal supply ad libitum
- Acclimation period: During the acclimation period of at least 5 days, the animals were kept under the same controlled environment conditions as during the experimental period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.1 – 23.9°C
- Humidity (%): 31 - 68%
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Route of administration:
oral: gavage
Vehicle:
methylcellulose
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 1 % Methyl cellulose
- Justification for choice of vehicle: The selection of the vehicle was made during trial formulations with the test item. In order of preference, recommended vehicles were: distilled water, 0.5 or 1% methylcellulose, PEG 400, oil (corn) and dimethyl sulfoxide. 1% methylcellulose was appropriate for treatment; therefore, it was found to be a suitable vehicle for formulations.
- Lot/batch no. (if required): S11183

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Per the guideline, a starting dose of 300 mg/kg bw was selected as there was no information available on the substance.
Doses:
300 mg/kg bw (Group 1 and 2), 2000 mg/kg bw (Group 3 and 4)
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
Morbidity and mortality: twice daily (at the beginning and end of each working day)
Clinical observations: after dosing at least once during the first 30 minutes, then 1, 2, 3, 4 and 6 hours after the treatment and once each day for 14 consecutive days thereafter or until death. Individual observations were performed on the skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The time of death was recorded as precisely as possible.
Body weights: Days -1 (prior to removal of food), 0 (prior to administration), 7 and 14 with a precision of 1 g.

- Necropsy of survivors performed: Yes; All animals were subjected to a necropsy and a macroscopic examination. exsanguinated after verification of narcosis following an injection of sodium pentobarbital. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross macroscopic changes were recorded for each animal.

Statistics:
No statistical evaluation was performed in this study
Preliminary study:
Not applicable
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred in the study during the 14-day observation period at the dose levels of 300 or 2000 mg/kg bw.
Clinical signs:
other: All animals were symptom-free during the 14-day observation period.
Gross pathology:
There was no evidence of the macroscopic changes at dose levels of 300 or 2000 mg/kg bw at necropsy.
Other findings:
Initially, three females (Group 1) were treated at a dose level of 300 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level. No mortality was observed in the confirmatory group; therefore, one further group (Group 3) was treated at the dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 4) was treated at the same dose level. No mortality was observed in this group and the study was terminated.
Interpretation of results:
GHS criteria not met
Conclusions:
The LD50 in female Crl:WI Wistar rats is > 2000 mg/kg bw.
Executive summary:

In an acute oral toxicity test (20/071-001P), 4 groups of female Crl:WI Wistar rats (3/group) were administered the test item (92.4%) in 1% Methyl cellulose by oral gavage at 300 mg/kg bw (2 steps) and 2000 mg/kg bw (2 steps). Animals were observed for 14 days.


 


The LD50 was > 2000 mg/kg bw.


 


No mortality occurred in the study during the 14-day observation period at the dose levels of 300 or 2000 mg/kg bw. All animals were symptom-free during the 14-day observation period. There were no test item related body weight changes. Body weights were within the range commonly recorded for this strain and age. There was no evidence of the macroscopic changes at dose levels of 300 or 2000 mg/kg bw at necropsy.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The key study was the only study available and was assigned a Klimisch score of 1. The overall quality of the database is high.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
4 March 2021-April 26 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity: Fixed Dose Procedure)
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier) and lot/batch number of test material: Provided by Sponsor, Lot number: 91112Y
- Purity.: 92.4%


STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material:
The test substance was put into an air-tight container and stored in test substance storage room at room temperature(acceptable range:from 10℃-30℃) .
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: stability


- Treatment of test material prior to testing (e.g. warming, grinding): The test item was crushed and corrected for purity.




Species:
rat
Strain:
Crj: CD(SD)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Japan(Hino Breeding Center)
- Age at study initiation: nine weeks old at the application of the range-finding study and the main study
- Weight at study initiation: 223.0 g in the range-finding study and 244.8 and 233.4 g in the main study
- Housing: The animals were housed individually after the group allocation and housed with two or three animals per cage after test substance was removed in the main study. The animals were housed in the barrier-system animal rooms in stainless steel cages with mesh floor.
- Diet: Free access to pelleted die t(MF, lot# 201008 and 201214, Oriental Yeast)
- Water: Chlorinated water in which chloric level maintained from 3 to 5 ppm by adding sodium hypochlorite (Purelox) to Hita City supply water was used as drinking water (ad libitum)
- Acclimation period: six days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 25℃
- Humidity (%): 40 to 70%
- Air changes (per hr): 10 to15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light per day


Type of coverage:
occlusive
Vehicle:
water
Remarks:
Purified water
Details on dermal exposure:
TEST SITE
- Area of exposure: back of the trunk of the animal
- % coverage: 5*10 cm area
- Type of wrap if used: The nonwoven fabric was covered and fixed with an elastic adhesive bandage (Silkytex, lot number 10114202, ALCARE)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The elastic adhesive bandage and the nonwoven fabric were removed from the animal and the application site was wiped with an absorbent cotton containing purified water to remove the residual test substance.
- Time after start of exposure: After 24 hours of application,

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Range finding study: animal number 1 = 0.48312; Mian study: animal number 2 = 0.53044g and animal number3 = 0.50546g (2000 mg/kg bw)
- For solids, paste formed: The test substance was put on a nonwoven fabric (5x5cm, lot number 201509151, Ci Medical), moistened with 1mL of purified water and applied to the clipped area.

VEHICLE
- Amount(s) applied (volume or weight with unit): 1 mL
- Lot/batch no. (if required): PC201111 (Takasugi Pharma)




Duration of exposure:
24h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
Range-finding: 2000mg/kg bw; 1 animal
Main study: 2000mg/kg bw; 2 animals
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
In the range-finding study and main study, the animals were observed continuously for 10 minutes after the application and observed 30 minutes and three hours after the application. From the day after the application (1 day after the application) to 14 days after the Application, the animals were observed once in the mooring. One day after the application, the animals were observed after the elastic adhesive bandages and the nonwoven fabrics were removed. Body weights were measured on the application day (before application) and 7 and 14 days after the application with an electric balance (SARTORIUS) and body weight changes were calculated.

- Necropsy of survivors performed: The animals were subjected to a gross necropsy 14 days after the application after the general clinical observation. The animals were euthanized by bleeding from the abdominal aorta under isoflurane anesthesia. Application site, external surface of the body, all orifices, subcutis, cranial, thoracic, abdominal and pelvic cavities with their contents were observed.

Preliminary study:
No mortality or moribundity, no body-weight abnormalities, no macroscopic findings abnormalities occurred in the range-finding study at 2000 mg/kg.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No mortalities in the main study
Clinical signs:
other: Decreased spontaneous locomotion was observed in all animals from immediately after the application to three hours after the application but disappeared one day after the application and no abnormalities were observed in any animals thereafter.
Gross pathology:
No abnormalities in the main study
Interpretation of results:
GHS criteria not met
Conclusions:
In an acute dermal toxicity study in female Crj:CD(SD) rats, the LD50 was >2000 mg/kg bw.
Executive summary:

In an acute dermal toxicity test (A18-0086), 2 groups of female Crj:CD(SD) rats (1/group in range finding; 2/group in main study) were dermally exposed (occlusive) to the test item in purified water at 2000 mg/kg bw. Animals were observed for 14 days.



Dermal LD50 (Females) = >2000 mg/kg bw



The test item applied at a dose of 2000 mg/kg bw did not cause death of any animals. There were no effects on body weight throughout the study. Decreased spontaneous locomotion was observed in all animals from immediately after the application to three hours after the application but disappeared one day after the application and no abnormalities were observed in any animals thereafter. No macroscopic changes were diagnosed during pathological examination.


 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The key study was the only study available and was assigned a Klimisch score of 1. The overall quality of the database is high.

Additional information

Acute oral toxicity


There is one in vivo study in rats available.


 


In an acute oral toxicity test (OECD 423/GLP), 4 groups of female Crl:WI Wistar rats (3/group) were administered the test item (92.4%) in 1% Methyl cellulose by oral gavage at 300 mg/kg bw (2 steps) and 2000 mg/kg bw (2 steps). Animals were observed for 14 days. No mortality occurred in the study during the 14-day observation period at the dose levels of 300 or 2000 mg/kg bw. All animals were symptom-free during the 14-day observation period. There were no test item related body weight changes. Body weights were within the range commonly recorded for this strain and age. There was no evidence of the macroscopic changes at dose levels of 300 or 2000 mg/kg bw at necropsy. The LD50 was > 2000 mg/kg bw.


 


Acute dermal toxicity


There is one in vivo study in rats available.


 


In an acute dermal toxicity test (OECD 402/GLP), 2 groups of female Crj:CD(SD) rats (1/group in range finding; 2/group in main study) were dermally exposed (occlusive) to the test item in purified water at 2000 mg/kg bw. Animals were observed for 14 days. The test item applied at a dose of 2000 mg/kg bw did not cause death of any animals. There were no effects on body weight throughout the study. Decreased spontaneous locomotion was observed in all animals from immediately after the application to three hours after the application but disappeared one day after the application and no abnormalities were observed in any animals thereafter. No macroscopic changes were diagnosed during pathological examination. The LD50 was >2000 mg/kg bw.


 

Justification for classification or non-classification

Based on the available information in the dossier, the test item does not need to be classified for acute toxicity or specific target organ toxicity - single exposure when the criteria outlined in CLP Regulation (Annex I of 1272/2008/EC) are applied.