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Administrative data

Description of key information

Oral:

A GLP compliant study was conducted to determine acute oral toxicity of test item according to OECD 423. The LD50 cut off value of test item defined as 5000 mg/kg bw or unclassified since no mortalities or moribundities occurred at 2000 mg/kg bw.

Inhalation:

The acute inhalation toxicity of test after being snout only administrated to SD rats for 4 hours was assessed at the maximum attainable concentration of 2270 mg/m3 based on OECD 403. The acute inhalation LC50 in SD rats for test item was more than 2270 mg/m3 (2.27 mg/L).

Dermal:

The acute dermal toxicity of test item in Sprague Dawley rats was assessed based on OECD 402. The acute dermal LD50 in rats for test item was estimated to be more than 2000 mg/kg b.w..

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:
From 2017-09-05 to 2017-11-10
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)
Version / remarks:
2001
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
Lot number: 707001
Purity: 98.6%
Appearance: Colourless tranparent liquid
Storage conditions: Refrigerator (1-10°C) in an air-tight container
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
Source: Charles River Laboratories, Japan (Hino Breeding Center)
Age of animals on receipt: Seven weeks
Quarantine/acclimatisation period: Seven days
Age at time of administration: Eight weeks (first and second step; 300 mg/kg bw); Nine weeks (third and fourth step; 2000 mg/kg bw).
Body weight at time of administration: 189.6-195.0g (first step; 300 mg/kg bw dosing); 188.1-195.1g (second step; 300 mg/kg bw dosing); 195.7-203.8g (third step; 2000 mg/kg bw dosing); 186.8-192.4g (fourth step; 2000 mg/kg bw)
Temperature: 21-25°C
Relative humidity: 40-70%
Air changes: 10-15/hour
Photoperiod: 12 hours light/dark cycle
Caging: Stainless steel cages with mesh floor
Housing: Up to 3/cage during acclimatisation; Individual until dosing
Feed: Pellet diet (MF; Oriental Yeast); ad libitum
Water: Chlorinated water (chlorine at 3-5 ppm) by adding sodium hypochlorite to Hita City mains supply; ad libitum
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
Concentration of dosing formulation: 3.00% w/v (300 mg/kg bw); 20.0% w/v (2000 mg/kg bw)
Dosing volume: 10 mL/kg bw
Rationale for dosing: No toxicity data available therefore first step was 300 mg/kg bw
Animals fasted for 18-19 hours before administration and for 3-4 hours after administration
Dosing based on body weight measured on the day of dosing
Doses:
300 mg/kg bw (first and second step)
2000 mg/kg bw (third and fourth step)
No. of animals per sex per dose:
Three females/dose for each step
Control animals:
no
Details on study design:
Animals dosed with 300 mg/kg bw for the first and second steps were observed continuously for 10 minutes after dosing, 30 minutes and 3 hours after administration and then daily from 1 to 14 days after administration.
Animals dosed with 2000 mg/kg bw for the third and fourth steps were observed continuously for 10 minutes after dosing, 30 minutes, 1, 3 and 5 hours after administration and once on the morning after dosing. Additional observations in the evening were performed the day after dosing for the thirs step and for the first two days after dosing for the fourth step.

Body weights were measured before administration, then 1, 7 and 14 days after dosing.
Animals were subjected to a gross necropsy 14 days after dosing.
Animals were euthanised by bleeding from the abdominal aorta under isoflurane anaesthesia. External surface of the body, all orifices, subcutis, cranial, thoracic, abdominal and pelvic cavities with their contents were observed.
Statistics:
Not required.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Mortality:
No mortalities or moribundities for any treatment.
Clinical signs:
At 300 mg/kg bw, mucus stools were observed in four animals 30 minutes or 3 hours after dosing. This finding was attibuted to the effect of the vehicle since this was not dose dependent and historical data show this has occured in 38/135 females following administration of olive oil to starved Crl:CD (SD) rats. A decreased stool volume was observed in one animal which was not considered to be an adverse effect since no changes were observed in the body weight. No other abnormalities were observed.

At 2000 mg/kg bw, decreased spontaneous locomotion, decreased respiratory rate and incomplete eyelid opening were observed in all six animals from 30 minutes to 1 hour after dosing. Salivation was observed in one animal 30 minutes after administration. Restlessness was noted in two animals and lacrimation in one animal 1 hour after dosing. One day after administration, decreased spontaneous locomotion in all six animals and decreased respiratory rate and incomplete eyelid opening in three animals were observed. Decreased spontaneous locomotion was noted in three animals two days after administration; no abnormalities were noted in the remaining three animals. No other abnormalities were observed three days after dosing or for the remaining observation period.
Body weight:
A decrease in body weight of 1.4g was noted for one animal dosed with 300 mg/kg bw 14 days after dosing (when compared to the body weight recorded at 7 days) which was considered to be a physiological fluctuation. A decrease tendency in body weight gain (weight gains of 2.4 and 4.6g) was observed in two animals (compared to gains of 6.0 to 10g for the remaining 4 animals) dosed with 2000 mg/kg bw one day after administration.

No other abnormalities were noted.
Gross pathology:
No abnormalities were observed at 300 or 2000 mg/kg bw.
Other findings:
None

Clinical signs

Clinical sign

Dose group

300 mg/kg bw

(first step)

300 mg/kg bw

(second step)

2000 mg/kg bw

(third step)

 

2000 mg/kg bw (fourth step)

 

Salivation

0/3

0/3

0/3

1/3

Mucus stool

1/3

3/3

0/3

0/3

Decreased stool volume

0/3

1/3

0/3

0/3

Decreased spontaneous locomotion

0/3

0/3

3/3

3/3

Decreased respiratory rate

0/3

0/3

3/3

3/3

Incomplete eyelid opening

0/3

0/3

3/3

3/3

Restlessness

0/3

0/3

1/3

1/3

Lacrimation

0/3

0/3

0/3

1/3

 

Interpretation of results:
GHS criteria not met
Remarks:
Not classified
Conclusions:
The LD50 cut off value of test item defined as 5000 mg/kg bw or unclassified since no mortalities or moribundities occurred at 2000 mg/kg bw.
Executive summary:

A GLP compliant study was conducted to determine acute oral toxicity of test item according to OECD 423. Three female Crl: CD (SD) rats were initially administered with 300 mg/kg bw. Since no mortalities or moribundities were observed, a further group of three females were dosed with 300 mg/kg bw. Since the findings in this additonal group were consistent with the first treatment group, the dose was increased to 2000 mg/kg bw for a further group of three females. Since no mortalities or moribundities were observed in this group, a further three females were dosed with 2000 mg/kg bw. Clinical signs were observed continuously for the first 10 minutes, then at longer intervals up to 5 hours after dosing and at least once daily up to the end of the observation period of 14 days following test substance administration. Body weight was recorded throughout. The animals were then subject to gross necropsy. External surface of the body, all orifices, subcutis, cranial, thoracic, abdominal and pelvic cavities with their contents were examined. No mortalities or moribundities were observed. A slight decrease in body weight gain was noted in two animals dosed with 2000 mg/kg bw one day after administration. There were no clinical signs at 300 mg/kg bw. At 2000 mg/kg bw, decreased spontaneous locomotion was observed in all six animals from 30 minutes after dosing which persisted until one or two days after administration. Decreased respiratory rate and incomplete eyelid opening was also noted in all six animals within 1 hour of dosing which persisted until the following day for only the second group dosed with 2000 mg/kg bw. Salivation was observed in one animal 30 minutes after administration. Restlessness was noted in two animals and lacrimation in one animal 1 hour after dosing. No other abnormalities were observed three days after dosing or for the remaining observation period for the higher dose animals. No abnormalities were noted during gross necropsy. The LD50 cut off value of test item defined as 5000 mg/kg bw or unclassified since no mortalities or moribundities occurred at 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
A modern, GLP- and guideline-compliant study is available for the submission substance.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2019-07-26 to 2019-08-09
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
Version / remarks:
2009
Deviations:
yes
Remarks:
The temperature and relative humidity of housing room have small deviations, not affect the quality and integrity of the study
GLP compliance:
yes (incl. QA statement)
Test type:
fixed concentration procedure
Limit test:
yes
Specific details on test material used for the study:
Batch No.: 707001
Purity: 98.5%
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: SPF (Beijing) Biotechnology Co., Ltd.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 57~64 days
- Weight at study initiation: 330~372g for male rats and 234~239g for female rats
- Fasting period before study: not fasted
- Housing: Animals were raised in suspended, stainless steel cages (L32.0cm ×W28.0cm×H20.0cm) on cage racks (L167.0cm×W70.0cm×H171.0cm). There were 10 cages per layer, and 4 layers per rack. Animals were housed individually after exposure.
- Diet: ad libitum except restraining period
- Water: ad libitum except restraining period
- Acclimation period: 15 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.5°C ~25.9°C
- Humidity (%): 50% ~72%
- Photoperiod: 12 hour light and 12 hour dark
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
air
Mass median aerodynamic diameter (MMAD):
>= 1.16 - <= 1.31 µm
Geometric standard deviation (GSD):
>= 2.75 - <= 2.84
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: HOPE-MED 8052Hdynamic snout-only aerosol inhalation instrument (Equipment No. W-23)
- Exposure method:
Before exposure, each rat was restrained in a confined transparent polyacrylic tube. The exposure tubes wereinstalled in the portholes of the inhalation chamber and the chamber was sealed up. Filtered and compressed air was mixed with quantitative test item and aerosol was sent to exposure chamber (0.04m3). The test item moving speed and exposure airflow rate had been adjusted. The aerosol had been continuously generated from generation system on the top of the chamber with an aerosol producer. A slight negative pressure was maintained in outer plenum of chamber to prevent leakage of the test substance into the surrounding area. The exhausted air was removed from the outlet at the bottom of the chamber to absorption unit.
- Particle size distribution determination: Twice, at the beginning and at the end of exposure using Aerosol Instrument for Aerodynamic Particle Sizer® (APS™) 3321

TEST ATMOSPHERE
- Brief description of analytical method used: Actual concentration at the animals' breathing zone was determined by using filter gravimetric analysis. Samples were collected by usingfiber filter fixed in filter holder attached to ESA-3Z auto-sampling device, on which airflow(1 L/min), time (5 min) and volume (5 L) was set. The mass collected filter paper was weighed before (M1) and after collection (M2). The collected mass was calculated by the following formula: (M) = M2-M1. Actual concentration (C, mg/m3) = the collected mass (M, mg)/ the collected volume (V, L) ×1000 L/m3. Frequency of determination was aboutonce per hour.
Analytical verification of test atmosphere concentrations:
yes
Duration of exposure:
4 h
Concentrations:
Maximum attainable concentration: 2270 mg/m3
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:
Clinical observations were recorded once during the exposure and twice with more than 30 minutes interval after exposure on the exposure day and then once daily for up to end observation.
The animals were weighed in the first 24 hours after arrival, on the day of exposure prior to exposure (day 0), and on day 1, day 3, day 7 and day 14.
- Necropsy of survivors performed: yes, All surviving animals at the end of the study and that which died during the test were subjected to gross necropsy.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 2 270 mg/m³ air (analytical)
Based on:
test mat.
Exp. duration:
4 h
Mortality:
One male animal was found dead during the exposure on the exposure day, the mortality was 17% (1/6).
Clinical signs:
other:
Body weight:
The body weights of three animals decreased on the Day 1 and increased on the Day 3 after exposure.
The body weight of one animal showed no change on the Day 1 and increased on the Day 3 after exposure.
The body weight of one animal showed increased trendduring the observation period.
Gross pathology:
No abnormalities were found in female and male animals at the gross necropsy.

 Calculated MMAD, GSD and Inhalable Fraction

Actual Concentration

Times

MMAD (µm)

GSD

<4 µm (mass%)

(mg/m3)

2270

First

1.16

2.75

66%

Second

1.31

2.84

63%

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute inhalation LC50 in SD rats for test item was more than 2270 mg/m3 (2.27 mg/L).
Executive summary:

The acute inhalation toxicity of test after being snout only administrated to SD rats for 4 hours was assessed at the maximum attainable concentration of 2270 mg/m3based on OECD 403.

One male animal was found dead during the exposure on the exposure day, the mortality was 17% (1/6).

One male animal showed tremor, dyspnea and convulsion, and one male animal and three female animals showed no abnormalities during the exposure.

The animals showed unkempt fur (4/5), blooding discharge around the nose (4/5), and secretions around the nose and mouth (2/5) on the first and second time observation after exposure on the exposure day.

One male animal showed unkempt fur, and one male animal and three female animals showed no abnormalities on the Day 1 after exposure. All surviving animals (5/5) showed no abnormalities on the Day 2-14 after exposure.

The body weights of three animals decreased on the Day 1 and increased on the Day 3 after exposure. The body weight of one animal showed no change on the Day 1 and increased on the Day 3 after exposure. The body weight of one animal showed increased trend during the observation period.

No abnormalities were found in female and male animals at the gross necropsy.

Based on the results, the acute inhalation LC50in SD rats for test item was more than 2270 mg/m3(2.27 mg/L).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
Value:
2 270 mg/m³
Quality of whole database:
A modern, GLP- and guideline-compliant study is available for the submission substance.

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:
From 2019-05-28 to 2019-06-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
2017
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
no
Specific details on test material used for the study:
Batch No.: 707001
Purity: 98.5%
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: SPF (Beijing) Biotechnology Co., Ltd.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 56-78 days
- Weight at study initiation: 240-280 g
- Fasting period before study: not fasted
- Housing: Animals were raised in suspended, stainless steel cages (L32.0cm × W60.0cm× H20.0cm) on cage racks (Ll99.0cm x W70.0cm x H 171.0 cm). There were 3 cages per layer and 4 layers per rack. Animals were housed individually during the exposure period and returned to group housing after the exposure period.
- Diet: SPF rodent maintenance feed, ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.9 - 25.0
- Humidity (%): 42 - 69
- Photoperiod: 12 hrs dark / 12 hrs light
Type of coverage:
not specified
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
Administration method:
- Fur Removing: Approximately 24 hours before the test, fur was removed from the dorsal area of the trunk of the test animals by clipping. Care was taken to avoid abrading the skin, which could alter its permeability. Clear area was about 40 cm2. Only animals with healthy and intact skin were used.
- Dosing Frequency: Each animal was dosed once.
- Dosing: The test item was applied evenly to the treatment area. The gauze was placed over the treatment area and wrapped with a piece of self-adhesive bandage. Shortly after dosing the dressings were examined to ensure that the animals cannot ingest the test item.
- Rest Test Item Removing: At the end of the exposure period after approximate 24 hours, residual test item was removed by cotton wool soaked in water.
The time intervals of each animal’s dosing were 2-5 days.


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): administered as supplied at a variable dose volume calculated in accordance with the design of dose and animal body weight
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
2
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical observations were performed once during the first 30 minutes and at 1, 2, 4 and 6 hours after application approximately and then once each day for 14 days.
Individual weights of animals were determined within 24 hours after arrival, at grouping, on Day 0 (day of dosing), Day 7 and Day 14.
- Necropsy of survivors performed: yes, Animals surviving to the end of the study were anesthetized by CO2 and bled by abdominal aorta to death.
Preliminary study:
No mortality occurred and no sign of systematic toxicity observed, the body weights showed growth trends, no abnormalities detected at all test levels.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths or moribund status were found
Clinical signs:
No abnormal symptoms were found
Body weight:
The body weights of two test animals with dose levels of 2000 mg/kg b.w. showed decrease trends during most of the observation periods.
Gross pathology:
No abnormalities were found in all test animals at necropsy.
Interpretation of results:
GHS criteria not met
Conclusions:
Based on the results, the acute dermal LD50 in rats for test item was estimated to be more than 2000 mg/kg b.w..
Executive summary:

The study was performed to assess the acute dermal toxicity of test item in Sprague Dawley rats based on OECD 402. A dose-range finding study with dose levels of 200, 1000, 2000 mg/kg b.w. and a main study with dose levels of 2000 mg/kg b.w. were carried out and five female animals were used.

No deaths or moribund status and no abnormal symptoms were found in all test animals from dosing until the end of the test. The body weights of two test animals with dose levels of 2000 mg/kg b.w. showed decrease trends during most of the observation periods. The body weights of the other test animals showed growth trends during most of the observation periods. No abnormalities were found in all test animals at necropsy.

Based on the results, the acute dermal LD50 in rats for test item was estimated to be more than 2000 mg/kg b.w..

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
A modern, GLP- and guideline-compliant study is available for the submission substance.

Additional information

Justification for classification or non-classification

Acute toxicity:

Oral:

Acute oral toxicity test, OECD 423: LD50: 5000 mg/kg

Inhalation:

Acute inhalation toxicity test, OECD 403: LC50: 2270 mg/m3 (2.27 mg/L).

Dermal:

Acute dermal test, OECD 402: LD50 >2000 mg/kg.

 

Therefore in accordance with Regulation (EC) No. 1272/2008 (amended by 286/2011) Table 3.1.1, this substance should be classified as Category 4 for acute inhalation and not classified for acute oral and dermal toxicity.

 

Specific target organtoxicity-single exposure:

Oral:

Acute oral toxicity test, OECD 423:

Mortality: no mortalities or moribundities occurred

Clinical observations:

There were no clinical signs at 300 mg/kg bw. At 2000 mg/kg bw, decreased spontaneous locomotion was observed in all six animals from 30 minutes after dosing which persisted until one or two days after administration. Decreased respiratory rate and incomplete eyelid opening was also noted in all six animals within 1 hour of dosing which persisted until the following day for only the second group dosed with 2000 mg/kg bw. Salivation was observed in one animal 30 minutes after administration. Restlessness was noted in two animals and lacrimation in one animal 1 hour after dosing. No other abnormalities were observed three days after dosing or for the remaining observation period for the higher dose animals.

Necropsy: No abnormalities were noted

 

Inhalation:

Acute inhalation toxicity test, OECD 403:

Mortality: One male animal was found dead during the exposure (1/6)

 

Clinical observations:

One male animal showed tremor, dyspnea and convulsion, and one male animal and three female animals showed no abnormalities during the exposure.

The animals showed unkempt fur (4/5), blooding discharge around the nose (4/5), and secretions around the nose and mouth (2/5) on the first and second time observation after exposure on the exposure day.

One male animal showed unkempt fur, and one male animal and three female animals showed no abnormalities on the Day 1 after exposure. All surviving animals (5/5) showed no abnormalities on the Day 2-14 after exposure.

Necropsy: No abnormalities were found

 

Dermal:

Acute dermal test, OECD 402:

Mortality: No deaths or moribund status were found

Clinical observations: No abnormal symptoms were found

Necropsy: No abnormalities were found

 

In accordance with Regulation (EC) No. 1272/2008 section 3.8.2.1.7., the effects observed are not considered as adverse effects that support classification, therefore this substance should be not classified.

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