Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 

Acute oral toxicity dose (LD50) of test chemical was considered based on experimental study conducted on rats, the LD50 value was considered to be >2000 mg/kg bw in rats. Thus, comparing this value with the criteria of CLP regulation, the test chemical cannot be classified for acute oral toxicity.

Acute Inhalation toxicity:

The acute Inhalation toxicity dose (LC50) for test chemical was considered based on the data available for the structurally and functionally similar read across chemicals. The study concluded that the LC50 value is >5 mg/L(>5000 mg/m3), for acute inhalation toxicity. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute inhalation toxicity.

Acute Dermal Toxicity:

Acute Dermal toxicity dose (LD50) for test chemical was considered based on different experimental studies conducted on rats and rabbits, the value was considered to be >2000 mg/kg bw in rats and rabbits. Thus, comparing this value with the criteria of CLP regulation, the test chemical cannot be classified for acute dermal toxicity.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
data is from experimental reports following standard procedures.
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
To determine the acute oral toxicity of test chemical in Sprague Dawley rats
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Specific details on test material used for the study:
- Substance Type: Organic
- Physical State: Liquid

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0001
- Expiration date of the lot/batch:July; 2021
- Manufacturing Date: July; 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item and prepared formulation(s) were stored at ambient temperature.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The dose of 300 mg/kg of test item was prepared by dilution of the test item in distilled water to obtain 30 mg/ml strength of solution. The formulation was prepared fresh on the day of dosing. The test item was administered in the dose volume of 10 ml/kg body weight. Actual preparation procedures are documented in the raw data. The dose of 2000 mg/kg of test item was administered undiluted as supplied by the Sponsor.

OTHER - Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: [yes/no] : yes
- Age at study initiation: Healthy young adult females were used. The age at study initiation was 8 to 12 weeks old
- Weight at study initiation: Body weights at the start : Female Mean : 199.48 g (= 100 %); Minimum : 194.3 g (- 2.60 %); Maximum : 207.2 g (+ 3.87 %)
- Fasting period before study: fasted rats (approximately 16 hours or more)
- Housing:polycarbonate cages.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum):Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period:All animals were acclimatized to laboratory conditions for minimum of 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.4 to 22.0 degree centigrade
- Humidity (%): 53.1% to 58.2%.
- Air changes (per hr): ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: From: 31-05-2017 To: 19-06-2017
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 300 mg/kg and 2000 mg/kg
- Amount of vehicle (if gavage):10 ml per kg of body weight

MAXIMUM DOSE VOLUME APPLIED: 10 ml per kg of body weight was considered the maximum volume which could be administered to a rat.

DOSAGE PREPARATION (if unusual): The dose of 300 mg/kg of test item was prepared by dilution of the test item in distilled water to obtain 30 mg/ml strength of solution.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:The doses were selected based on the available literature of the test item.
Doses:
Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg
No. of animals per sex per dose:
Three females were used at each step
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality and morbidity, until sacrifice.Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.
Body weights: Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.
Gross Pathology: Necropsy was performed on all animals at the end of the study period on day 15. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique.
Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.
Statistics:
Mean ± SD
Preliminary study:
not specified
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
No mortality was observed at all the tested concentrations at all times
Clinical signs:
No clinical signs and signs of toxicity were observed.
Body weight:
Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.32% and 12.11% respectively.
Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 2.91% and 11.68% respectively.
Group II Step I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.77% and 14.43% respectively.
Group II Step II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 2.80% and 12.50% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.
Other findings:
- Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.

Table 1: Summary of Clinical Signs of Toxicity and Mortality

Sprague Dawley Rat

Sex : Female

Group I :   

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

300

No clinical signs observed

3

1,2,3

Day 0 - Day 14

0/3

 

 

 

Group I :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

II

300

No clinical signs observed

3

4,5,6

Day 0 - Day 14

0/3

 

Group II :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

2000

No clinical signs observed

3

7,8,9

Day 0 - Day 14

0/3

 

Group II :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

II

2000

No clinical signs observed

3

10,11,

12

Day 0 - Day 14

0/3

 

 

 

Table 2: Mean Body Weight and Percent Body Weight Gain (g)

Sprague Dawley Rat

Sex : Female

Group I :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

300

Mean

199.03

205.60

3.32

223.10

8.51

12.11

± SD

3.23

1.90

2.28

2.48

0.63

2.11

 

 

Group I :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

II

300

Mean

203.43

209.37

2.91

227.20

8.52

11.68

± SD

4.60

5.75

0.75

5.64

0.37

0.43

 

 

Group II :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

2000

Mean

199.17

206.67

3.77

227.90

10.27

14.43

± SD

2.14

2.81

1.00

4.19

0.67

1.78

 

 

Group II :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

II

2000

Mean

196.30

201.80

2.80

220.83

9.44

12.50

± SD

2.17

2.61

0.20

1.40

1.13

0.94

 

Table 3: Summary of Gross Pathological Findings

Sprague Dawley Rat

Sex : Female

 

        Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

300

1 - 3

TS

No abnormality detected

 

 

        Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

300

4 - 6

TS

No abnormality detected

 

 

                    Group II :

 

Step No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

7 - 9

TS

No abnormality detected

 

 

                    Group II :

 

Step No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

2000

10 - 12

TS

No abnormality detected

 

                     TS = Terminal Sacrifice

 

                    

 

 

 

Interpretation of results:
other: Not classified
Conclusions:
The acute oral LD50 value for test chemical can be considered to be >2000mg/kg. Hence, it was concluded that the acute toxicity of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.
Executive summary:

The study was designed and conducted to determine the acute oral toxicity profile of test chemical in Sprague Dawley rats. The study was performed according to OECD 423 - Acute Toxic Class Method. Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Group I Step I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Group I Step II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. As no mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Group II Step I). Administration of the test item at 2000 mg/kg did not result in any signs of toxicity and mortality after the dosing. As no mortality were observed at 24 hours after the dosing, additional three female animals were treated with the higher dose of  2000 mg/kg of the test item (Group II Step II). Administration of the test item at 2000 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality or gross pathological abnormalities were observed in animals from 300 mg/kg and 2000 mg/kg dose groups. Therefore, the acute oral LD50 value for test chemical can be considered to be >2000mg/kg. Hence, it was concluded that the acute toxicity of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from experimental study report.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data of read across substances
Justification for type of information:
Data for the target chemical is summarized based on the structurally and functionally similar read across chemicals
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on two acute inhalation toxicity studies as- WoE 2 and WoE 3.
Acute inhalation toxicity test was carried out to study the effects of the test chemicals on rodents
GLP compliance:
not specified
Test type:
other: not specified
Limit test:
no
Species:
rat
Strain:
other: 1. Sprague-Dawley 2. not specified
Sex:
male/female
Details on test animals and environmental conditions:
1. TEST ANIMALS
- Source: Charles River UK
- Age at study initiation: 6 to 8 weeks
- Weight at study initiation: ca. 200 g
- Housing: Animals were kept in cages.
- Diet (e.g. ad libitum): Food was provided
- Water (e.g. ad libitum): water was provieded
2. not specified
Route of administration:
other: 1. inhalation: aerosol 2. inhalation
Type of inhalation exposure:
other: 1. whole body 2. not specified
Vehicle:
other: 1. clean air 2. not specified
Mass median aerodynamic diameter (MMAD):
5.8 µm
Geometric standard deviation (GSD):
2.23
Remark on MMAD/GSD:
1. Sampling with a Marple cascade impactor 35 minutes after the start of exposure.
2. not specified
Details on inhalation exposure:
1. GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: exposure chamber
- Exposure chamber volume: 120 L
- Method of holding animals in test chamber: Animals were held in 10 separate animal compartments.
- Source and rate of air: Respirable aerosol in clean dried air produced by aerosol generator (equilibration period 9 min),
- Temperature, humidity, pressure in air chamber: supply pressure 15 l/min, flow rate in the exposure chamber: 0.8 ml/min per pump, total flow rate 1.6 ml/min.

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution:
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): mass median aerodynamic diameter (MMAD): 5.8 µm, geometric standard deviation: 2.23, 59.5% in the respirable range (< 7 µm).

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Analytical determination, sampling through a gas absorption trap, 3 samples were taken during the exposure period
2. not specified
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
1 h
Remarks on duration:
not specified
Concentrations:
1. 7.38 mg/l +- 0.131 (nominal 58 mg/l or 7380 mg/m3)
2. 4000 ppm
No. of animals per sex per dose:
1. 5 males, 5 females per group
2. not specified
Control animals:
other: 1. yes, clean air only 2. not specified
Details on study design:
1. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical signs: continuously during exposure, twice daily during the observation period.
Body weights: daily. Food and water consumption per cage.
- Necropsy of survivors performed: yes
- Other examinations performed: Macroscopic organ examination at necropsy, lung weights.
2. not specified
Statistics:
1. not specified
2. not specified
Preliminary study:
1. not specified
2. not specified
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 7 380 mg/m³ air (analytical)
Based on:
test mat.
Exp. duration:
1 h
Remarks on result:
other: No mortality was observed
Sex:
not specified
Dose descriptor:
LC50
Effect level:
> 4 000 ppm
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
1. No mortality was observed at 7380 mg/m3.
2. No mortality was observed
Clinical signs:
1. During observation period, immediately after removal from the exposure chamber: exaggerated respiratory movements and matted fur (all rats), wet fur around the snout and jaws (2 males) and clear discharge from the eyes (3 females), brown staining around the snout and jaws (1 female) were observed. All symptoms were reversible within 24 hours.
2. not specified
Body weight:
1. Slightly reduced body weight and food consumption on day 1, thereafter comparable to control. Water consumption not affected.
2. not specified
Gross pathology:
1. At necropsy there were no macroscopic abnormalities in the rats.
2. not specified
Other findings:
1. Lung to body weight ratio were comparable between test and control animals.
2. not specified
Interpretation of results:
other: Not classified
Conclusions:
According to CLP regulation, the test chemical test chemical cannot be classified for acute inhalation toxicity, as the LC50 value is >5 mg/L (>5000 mg/m3).
Executive summary:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute inhalation toxicity of the test chemical.The studies are as mentioned below:

1. Acute inhalation toxicity study of test chemical was conducted in 10 male and female Sprague-Dawley rats at the dose concentration of 7380 mg/m3 (7.38 mg/l). The exposure was given to whole body, respirable aerosol in clean dried air produced by aerosol generator (equilibration period 9 min), and exposure chamber 120 l with 10 separate animal compartments. The control animals received clean air only. Animals were observed for - clinical signs: continuously during exposure, twice daily during the observation period; body weights: daily; Food and water consumption per cage. Necropsy of survivors was performed. Macroscopic organ examination at necropsy and lung weights was done. No mortality was observed at 7380 mg/m3.During observation period, immediately after removal from the exposure chamber: exaggerated respiratory movements and matted fur (all rats), wet fur around the snout and jaws (2 males) and clear discharge from the eyes (3 females), brown staining around the snout and jaws (1 female) were observed. All symptoms were reversible within 24 hours. Slightly reduced body weight and food consumption on day 1, thereafter comparable to control. Water consumption not affected. At necropsy there were no macroscopic abnormalities in the rats. Lung to body weight ratio were comparable between test and control animals. Therefore, LC50 value was considered to be >7380 mg/m3, when 10 male and female Sprague-Dawley rats were treated with test chemical via inhalation route by aerosol to whole body exposure for 1 hour.

2. The acute inhalation toxicity study was conducted by using test chemical in rats at the concentration of 4000 ppm. No mortality was observed at 4000 ppm. Hence, LC50 value was considered to be >4000 ppm, when rats were exposed to test chemical via inhalation route.

Thus, based on the above summarised studies, test chemical and it’s structurally and functionally similar read across substances, it can be concluded that LC50 value is >5 mg/L. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute inhalation toxicity.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
5 000 mg/m³
Quality of whole database:
Data is Klimisch 2 and from authoritative database.

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
data is from experimental reports following standard procedures
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
To determine the acute dermal toxicity of test chemical in Sprague Dawley rats.
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0001
- Expiration date of the lot/batch:July; 2021
- Manufacturing Date: July; 2016

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Test Item and prepared formulation(s) were stored at ambient temperature.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was used undiluted and as supplied by the Sponsor.

OTHER - Safety Precautions: Safety precautions included use of protective clothing, gloves, masks and eye protection (glasses).
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Sex: Male and Female
- Source: National Institute of Biosciences, Pune.
- Age at study initiation: Young adult male and female rats aged between 8 – 12 weeks
- Weight at study initiation: Body weights at the start : Male - Mean: 258.62 g (= 100 %); Minimum : 251.4 g (- 2.79 %); Maximum : 266.4 g (+ 3.01 %)
Total No. of animals : 5
Female- Mean : 230.04 g (= 100 %); Minimum : 224.3 g (- 2.50 %); Maximum : 238.4 g (+ 3.63 %)
Total No. of animals : 5
- Housing:individually housed in polycarbonate cages with paddy husk as bedding
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences, Pune, was provided ad libitum from individual feeders.
- Water (e.g. ad libitum):Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period:All animals were acclimatized to laboratory conditions for minimum of 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.4 to 22.0 degree centigrade
- Humidity (%): 55.4% to 58.8%.
- Air changes (per hr): ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: From: 06-06-2017 To: 21-06-2017
Type of coverage:
occlusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal surface and sides from scapular to pelvic area
- % coverage: approximately 10% of the total body surface area
- Type of wrap if used: using a porous gauze dressing and non irritating tape
REMOVAL OF TEST SUBSTANCE

- Washing (if done): yes, the wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg
Duration of exposure:
24 hrs
Doses:
2000 mgkg
No. of animals per sex per dose:
A single dose of 2000 mg of the test item per kilogram of body weight was administered to ten rats (five males and five females).

Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality, until sacrifice. Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.
Evaluation of Dermal Reaction: Dermal reaction was observed daily for study period of 14 days.
Body weights: Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.
Gross Pathology: Necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).
Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.
Statistics:
not specified
Preliminary study:
not specified
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no other details available
Mortality:
Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. All animals survived through the study period of 14 days.
Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. All animals survived through the study period of 14 days
Clinical signs:
Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Body weight:
Sex : Male Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 9.46% and 17.48% respectively.
Sex : Female Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.84% and 9.86% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group
Other findings:
Evaluation of Dermal Reaction - Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

Table 1: Summary of Clinical Signs of Toxicity and Mortality

Sprague Dawley Rat

Sex : Male

Group

 No.

Dose mg/kg

                            Observed Signs

Total Number of

Animals

 

Animal Nos.

Period of signs in days

 From - to

 

Mortality

I

2000

No clinical signs observed

5

1 - 5

Day 0 - Day 14

0/5

 Sex : Female

 

Group

 No.

Dose mg/kg

                            Observed Signs

Total Number of

Animals

 

Animal Nos.

Period of signs in days

 From - to

 

Mortality

I

2000

No clinical signs observed

5

6 - 10

Day 0 - Day 14

0/5

Table 2: Summary of Evaluation of Dermal Reaction

Sprague Dawley Rat

Sex : Male 

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

 

Animal Nos.

Period of signs

in days

 From - to

 

Mortality

I

2000

No dermal reaction observed

5

1 - 5

Day 0 - Day 14

0/5

 

Sex : Female

 

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

 

Animal Nos.

Period of signs

in days

 From - to

 

Mortality

I

2000

No dermal reaction observed

5

6 - 10

Day 0 - Day 14

0/5

 

Table 3: Mean Body Weight and Percent Body Weight Gain (g)

Sprague Dawley Rat

Sex : Male

Group No.

Dose

(mg/kg body weight)

 

Body weight Day 0

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

2000

Mean

258.62

283.10

9.46

303.84

7.33

17.48

± SD

5.70

6.54

0.57

7.88

1.51

1.35

Sex : Female

Group No.

Dose

(mg/kg body weight)

 

Body weight Day 0

Body weight Day 7

% body weight gain

day 0-7

Body weight Day 14

% body weight gain

day 7- 14

% body weight gain

day 0- 14

I

2000

Mean

230.04

241.18

4.84

252.74

4.79

9.86

± SD

5.50

6.35

0.50

7.32

0.44

0.83

 

Table 4: Summary of Gross Pathological Findings

Sprague Dawley Rat

Sex : Male

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

1 - 5

TS

No abnormality detected

Sex : Female

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

6 - 10

TS

No abnormality detected

                     TS = Terminal Sacrifice

 

Interpretation of results:
other: Not classified
Conclusions:
The acute dermal LD50 value for test chemical can be considered to be >2000mg/kg bw. Hence, it was concluded that the acute toxicity of test chemical, when applied via dermal route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.
Executive summary:

The study was designed and conducted to determine the acute dermal toxicity profile of test chemical in Sprague Dawley rats. The study was performed according to OECD 402 Guidelines. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. No mortality or gross pathological abnormalities were observed in animals at the limit dose 2000 mg/kg bw. Therefore, the acute dermal LD50 value for test chemical can be considered to be >2000mg/kg bw. Hence, it was concluded that the acute toxicity of test chemical, when applied via dermal route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from experimental study report.

Additional information

Acute oral toxicity:

In different experimental studies, test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats and rabbits for test chemical. The studies are summarized as below –

The study was designed and conducted to determine the acute oral toxicity profile of test chemical in Sprague Dawley rats. The study was performed according to OECD 423 - Acute Toxic Class Method. Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Group I Step I). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Group I Step II). Administration of the test item at 300 mg/kg did not result in any signs of toxicity and mortality after the dosing. As no mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Group II Step I). Administration of the test item at 2000 mg/kg did not result in any signs of toxicity and mortality after the dosing. As no mortality were observed at 24 hours after the dosing, additional three female animals were treated with the higher dose of  2000 mg/kg of the test item (Group II Step II). Administration of the test item at 2000 mg/kg did not result in any signs of toxicity and mortality after the dosing. No mortality or gross pathological abnormalities were observed in animals from 300 mg/kg and 2000 mg/kg dose groups. Therefore, the acute oral LD50 value for test chemical Ethyl Butyrate (CAS No. 105-54-4) can be considered to be >2000mg/kg. Hence, it was concluded that the acute toxicity of test chemical when administered via oral route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.

The above experimental study is supported with the study mentioned in different sources, for the test chemical . Acute oral toxicity study was conducted in groups of 10 male and female Osborne-Mendel rats at the dose concentration range of 12210-13940 mg/kg bw. The test chemical (undiluted) was administered via oral gavage route. All animals were maintained under close observation for recording toxic signs and time of death for 14 days. Such observation was continued until animals appeared normal and showed weight gain. LD50’s were computed by the method of Litchfield & Wilcoxon (1949). Death occurred within 4 to 18 hours. Depression showed within a few minutes, with coma on higher doses. Therefore, LD50 value was considered to be 13050 mg/kg bw, with 95% confidence limits of 12210-13940 mg/kg bw, when groups of 10 male and female Osborne-Mendel rats were treated with test chemical via oral gavage route.

Both the above experimental studies are further supported with the study mentioned.The acute oral toxicity study was conducted in 10-35 male and female rabbits at the dose concentration of 5228 mg/kg bw. The test chemical was dissolved in saline and administered via oral gavage route. 50% mortality was observed in treated animals. Therefore, LD50 value was considered to be 5228 mg/kg bw, when 10-35 male and female rabbits were treated with test chemical via oral gavage route.

Thus, based on the above experimental studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:

Data available for the structurally and functionally similar read across chemicals has been reviewed to determine the acute inhalation toxicity of the test chemical.The studies are as mentioned below:

1. Acute inhalation toxicity study of test chemical was conducted in 10 male and female Sprague-Dawley rats at the dose concentration of 7380 mg/m3 (7.38 mg/l). The exposure was given to whole body, respirable aerosol in clean dried air produced by aerosol generator (equilibration period 9 min), and exposure chamber 120 l with 10 separate animal compartments. The control animals received clean air only. Animals were observed for - clinical signs: continuously during exposure, twice daily during the observation period; body weights: daily; Food and water consumption per cage. Necropsy of survivors was performed. Macroscopic organ examination at necropsy and lung weights was done. No mortality was observed at 7380 mg/m3.During observation period, immediately after removal from the exposure chamber: exaggerated respiratory movements and matted fur (all rats), wet fur around the snout and jaws (2 males) and clear discharge from the eyes (3 females), brown staining around the snout and jaws (1 female) were observed. All symptoms were reversible within 24 hours. Slightly reduced body weight and food consumption on day 1, thereafter comparable to control. Water consumption not affected. At necropsy there were no macroscopic abnormalities in the rats. Lung to body weight ratio were comparable between test and control animals. Therefore, LC50 value was considered to be >7380 mg/m3, when 10 male and female Sprague-Dawley rats were treated with test chemical via inhalation route by aerosol to whole body exposure for 1 hour.

2. The acute inhalation toxicity study was conducted by using test chemical in rats at the concentration of 4000 ppm. No mortality was observed at 4000 ppm. Hence, LC50 value was considered to be >4000 ppm, when rats were exposed to test chemical via inhalation route.

Thus, based on the above summarised studies, test chemical and it’s structurally and functionally similar read across substances, it can be concluded that LC50 value is >5 mg/L. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute inhalation toxicity.

Acute Dermal toxicity:

In different experimental studies, test chemical has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments in rodents, i.e. most commonly in rats and rabbits for test chemical. The studies are summarized as below –

The study was designed and conducted to determine the acute dermal toxicity profile of test chemical in Sprague Dawley rats. The study was performed according to OECD 402 Guidelines. The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. No mortality or gross pathological abnormalities were observed in animals at the limit dose 2000 mg/kg bw. Therefore, the acute dermal LD50 value for test chemical can be considered to be >2000mg/kg bw. Hence, it was concluded that the acute toxicity of test chemical, when applied via dermal route in Sprague Dawley rats falls into the “Category Not classified” criteria of CLP.

The above experimental study is supported with the study mentioned in publication, handbook and authoritative databases, for the test chemical. The acute dermal toxicity study was conducted in rabbits at the dose concentration of 2000 mg/kg bw. No mortality was observed at 2000 mg/kg bw. Therefore, LD50 value was considered to be >2000 mg/kg bw, when rabbits were treated with test chemical by dermal application.

Thus, based on the above experimental studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation,test chemical cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above experimental studies on test chemical and it’s structurally and functionally similar read across substances, it can be concluded that LD50 value is >2000 mg/kg bw, for acute oral and acute dermal toxicity; and LC50 value is >5 mg/L, for acute inhalation toxicity. Thus, comparing these values with the criteria of CLP regulation, test chemical cannot be classified for acute oral, acute dermal and acute inhalation toxicity.