N-methylformanilide

Endpoint summary

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on different studies conducted on rats for the test chemical. The LD50 value is 800 mg/kg bw. The study concluded that LD50 value is between 300-2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 0.01995164 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on different studies conducted on rats and rabbits for the test chemical. The studies concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given 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:

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 from various test chemicals

Justification for type of information:

Data is summarized based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on 3 acute oral toxicity studies as - WoE 2, WoE 3 and WoE 4.
Acute Oral 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. not specified 2. Sprague-Dawley 3. Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

1. not specified
2. TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Female rats of the age of approximately 8 to 12 weeks old were used at the commencement of its dosing.
- Weight at study initiation: Body weight range was 197.2 to 203.7 grams.
Body weights at the start : Female Mean: 200.23 g (= 100 %); Minimum : 197.2 g (- 1.51 %); Maximum : 203.7 g (+ 1.73 %)
- Identification: Each female rat was individually identified by the picric acid marking.
- Fasting period before study: Approximately 16 hours or more.
- Housing: The rats were housed in 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: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.1 to 22.1 degree centigrade.
- Humidity (%): 56.6% to 60.1%.
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 08-08-2017 to 25-08-2017
3. TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Female rats of the age of approximately 8 to 12 weeks old were used at the commencement of its dosing and its weight were within ± 20% of the mean weight of any animal used for dosing.
- Weight at study initiation: Body weight range was 195.9 to 202.2 grams. Female Mean: 198.78 g (= 100 %); Minimum : 195.9 g (- 1.45 %); Maximum : 202.2 g (+ 1.72 %)
- Identification: Each female rat was individually identified by the picric acid marking.
- Fasting period before study: Approximately 16 hours or more.
- Housing: The rats were housed in 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: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.1 to 22.1 degree centigrade.
- Humidity (%): 56.6% to 60.1%.
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 08-08-2017 to 25-08-2017

Route of administration:

other: 1. oral: unspecified 2. oral: gavage 3. oral: gavage

Vehicle:

other: 1. unchanged (no vehicle) 2. 1% aqueous Tween 80 3. water

Details on oral exposure:

1. not specified
2. VEHICLE
- Concentration in vehicle: 300 mg/kg, 300 mg/kg and 2000 mg/kg
- MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.
- DOSAGE PREPARATION (if unusual): Test item was insoluble in water. Uniformly dispersed suspension of the test item was possible in 1% aqueous Tween 80.
3. VEHICLE
- Concentration in vehicle: 300 mg/kg, 300 mg/kg and 2000 mg/kg
- MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.

Doses:

1. Range of 100-1600 mg/kg bw
2. Dose Group I : 300 mg/kg
Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg
3. Dose Group I : 300 mg/kg
Dose Group I : 300 mg/kg
Dose Group II : 2000 mg/kg

No. of animals per sex per dose:

1. Total = 10
2. Three females were used at each step.
3. Three females were used at each step.

Control animals:

not specified

Details on study design:

1. - Duration of observation period following administration: 14 days
- Other examinations performed: Animals were observed for mortality, body weight change and clinical signs.
2. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- Other examinations performed: 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, found dead and sacrificed 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.
3. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- Other examinations performed: 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, found dead and sacrificed 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: Gross pathological examination revealed unabsorbed residual test item in distended stomach in found dead animals from 2000 mg/kg dose group. No gross pathological changes attributable to the treatment were observed in stomach and hence no histopathology was considered.

Statistics:

1. not specified
2. not specified
3. not specified

Preliminary study:

1. not specified
2. not specified
3. not specified

Sex:

not specified

Dose descriptor:

LD50

Effect level:

800 mg/kg bw

Based on:

test mat.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

1. Mortality was observed within 2 hrs to 1 day in treated animals at 800 mg/kg bw.
2. Group I Step I : Animals treated at the dose level of 300 mg/kg body weight: All animals survived through the study period of 14 days.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight: One animal died on day 1 after the dosing.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight: Three animals died at 1 hour after the dosing.
3. Group I Step I : Animals treated at the dose level of 300 mg/kg body weight: All animals survived through the study period of 14 days.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight: One animal died at 6 hours after the dosing.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight: Three animals died at 1 hour after the dosing.

Clinical signs:

1. Symptoms included weakness, roughening of the coat, cyanosis, tremors, darkening of the eyes, and lacrimation just before death.
2. Group I Step I : Animals treated at the dose level of 300 mg/kg body weight resulted in ptosis, distension, reduced locomotor activity, ataxic gait and tremors with onset at 10 minutes to 2 hours after the dosing. All animals survived through the study period of 14 days and were free of signs of toxicity from day 4 to day 5 after the dosing.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight resulted in ptosis, distension, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 4 hours after the dosing. One animal died on day 1 after the dosing. All surviving animals were free of signs of toxicity from day 3 to day 4 after the dosing.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing. Three animals died at 1 hour after the dosing.

Body weight:

1. Body weight change was observed in more than 6 animals.
2. Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 3.90% and 10.94% respectively.
Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.26% and 13.03% respectively.
Group II Step I (2000 mg/kg) - All animals died at 1 hour, hence, body weight gain could not be calculated.
3. Group I Step I (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.31% and 12.09% respectively.
Group I Step II (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.79% and 12.10% respectively.
Group II Step I (2000 mg/kg) - All animals died at 1 hour, hence, body weight gain could not be calculated.

Gross pathology:

1. not specified
2. Gross pathological examination did not reveal any abnormalities in found dead as well as terminally sacrificed animals from 300 mg/kg and 2000 mg/kg dose groups.
3. Gross pathological examination did not reveal any abnormalities in found dead animal and animals sacrificed terminally from 300 mg/kg dose group.
Gross pathological examination revealed stomach distended with unabsorbed residual test item in found dead animals from 2000 mg/kg dose group.

Other findings:

1. not specified
2. not specified
3. Group I Step I : Animals treated at the dose level of 300 mg/kg body weight resulted in distension, piloerection, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 2 hours after the dosing. All animals survived through the study period of 14 days and were free of signs of toxicity from day 4 to day 5 after the dosing.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and loss of righting reflex with onset at 30 minutes to 4 hours after the dosing. All surviving animals were free of signs oftoxicity from day 4 to day 5 after the dosing.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight resulted in reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

According to CLP regulation, the test chemical can be classified in "Category 4" for acute oral toxicity, as the LD50 value is in the range of 300-2000 mg/kg bw.

Executive summary:

In different studies, the given 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 for test chemical. The studies are summarized as below –

 

Acute oral toxicity study of the given test chemical was conducted in 10 rats at the dose concentration range of 100-1600 mg/kg bw. The given test chemical (undiluted) was administered via oral route. Animals were observed for mortality, body weight change and clinical signs for 14 days.

Mortality was observed within 2 hrs to 1 day in treated animals at 800 mg/kg bw. Symptoms included weakness, roughening of the coat, cyanosis, tremors, darkening of the eyes, and lacrimation just before death. Body weight change was observed in more than 6 animals.

Under the condition of this, the LD50 value was considered to be 800 mg/kg bw, when 10 rats were treated with the given test chemical via oral route.

 

The above study is supported with another study mentioned in study report for the test chemical. The reported study was designed and conducted to determine the acute oral toxicity profile of the given test chemical as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) in Sprague Dawley rats.

Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in ptosis, distension, reduced locomotor activity, ataxic gait and tremors with onset at 10 minutes to 2 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 (Step - II). Administration of the test item at 300 mg/kg resulted in ptosis, distension, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 4 hours after the dosing. One animal died on day 1 after the dosing. One 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 (Step - I). Administration of the test item at 2000 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing. Three animals died at 1 hour after the dosing.

All surviving animals from 300 mg/kg dose group exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in found dead as well as terminally sacrificed animals from 300 mg/kg and 2000 mg/kg dose groups.

Under the condition of this, the lethal concentration (LD50) value for acute oral toxicity test was considered in between 300-2000 mg/kg bw, when female Sprague Dawley rats were treated with the given test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

 

Both the above studies are further supported with the study mentioned in study report for the test chemical. The reported study was designed and conducted to determine the acute oral toxicity profile of the given test chemical as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) in Sprague Dawley rats.

Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 2 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 (Step - II). Administration of the test item at 300 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and loss of righting reflex with onset at 30 minutes to 4 hours after the dosing. One animal died at 6 hours after the dosing. One 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 (Step - I). Administration of the test item at 2000 mg/kg resulted in reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing. Three animals died at 1 hour after the dosing.

All surviving animals from 300 mg/kg dose group exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in found dead animal and animals sacrificed terminally from 300 mg/kg dose group. Gross pathological examination revealed stomach distended with unabsorbed residual test item in found dead animals from 2000 mg/kg dose group.

Hence, it was concluded that the lethal concentration (LD50) value for acute oral toxicity test was considered in between 300-2000 mg/kg bw, when female Sprague Dawley rats were treated with the given test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is between 300-2000 mg/kg bw. Thus, comparing this range with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute oral toxicity.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

800 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from database.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Endpoint conclusion

Quality of whole database:

Waiver

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

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 from various test chemicals

Justification for type of information:

Data is summarized based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on 3 acute dermal toxicity studies as - WoE 2, WoE 3 and WoE 4.
Acute dermal 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:

other: 1. rat 2. rat 3. rabbit

Strain:

other: 1. Sprague-Dawley 2. Sprague-Dawley 3. not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

1. TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Age at study initiation: Young adult male and female rats aged between 8 – 12 weeks were used.
- Weight at study initiation: The weight range of approximately 212.0 to 237.6 grams at initiation of dosing.
Body weights at the start : Male Mean: 233.08 g (= 100 %); Minimum : 228.7 g (- 1.88 %); Maximum : 237.6 g (+ 1.94 %)
Female Mean : 215.88 g (= 100 %); Minimum : 212.0 g (- 1.80 %); Maximum : 220.5 g (+ 2.14 %)
- Identification: Each rat was individually identified by the cage number.
- Housing: The rats were 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: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6 to 22.6 degree centigrade.
- Humidity (%): 55.1% to 59.7%.
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 09-08-2017 to 24-08-2017
2. TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Age at study initiation: Young adult male and female rats aged between 8 – 12 weeks were used.
- Weight at study initiation: The weight range of approximately 211.3 to 235.7 grams at initiation of dosing.
Body weights at the start : Male Mean: 233.06 g (= 100 %); Minimum : 228.8 g (- 1.83 %); Maximum : 235.7 g (+ 1.13 %)
Female Mean : 216.42 g (= 100 %); Minimum : 211.3 g (- 2.37 %); Maximum : 220.0 g (+ 1.65 %)
- Identification: Each rat was individually identified by the cage number.
- Housing: The rats were 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: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.6 to 22.6 degree centigrade.
- Humidity (%): 55.1% to 59.7%
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 09-08-2017 to 24-08-2017
3. not specified

Type of coverage:

other: 1. occlusive 2. occlusive 3. Dermal

Vehicle:

other: 1. water 2. water 3. not specified

Details on dermal exposure:

1. TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- For solids, paste formed: Yes
2. TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- For solids, paste formed: Yes
3. not specified

Duration of exposure:

1. 24 hours
2. 24 hours
3. not specified

Doses:

1. 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).
2. 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).
3. 5000 mg/kg bw

No. of animals per sex per dose:

1. 10 (5/sex).
2. 10 (5/sex).
3. not specified

Control animals:

not specified

Details on study design:

1. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- Other examinations performed: 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.
2. - Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- Other examinations performed: 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.
3. not specified

Statistics:

1. not specified
2. not specified
3. not specified

Preliminary study:

1. not specified
2. not specified
3. not specified

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality was observed

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

1. Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight: 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: All animals survived through the study period of 14 days.
2. Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg: All animals survived through the study period of 14 days.
Sex : Female Group I – Animal treated at the dose level of 2000 mg/kg: All animals survived through the study period of 14 days.
3. No mortality was observed at 5000 mg/kg bw

Clinical signs:

1. 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.
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.
2. 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.
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.
3. not specified

Body weight:

1. Sex : Male Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 10.39% and 21.25% respectively.
Sex : Female Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.74% and 12.52% respectively.
2. Sex : Male Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 10.37% and 21.31% respectively.
Sex : Female Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.21% and 9.81% respectively.
3. not specified

Gross pathology:

1. Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.
2. Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.
3. not specified

Other findings:

1. - Other observations: Evaluation of Dermal Reaction
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.
2. - Other observations: Evaluation of Dermal Reaction
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.
3. not specified

Interpretation of results:

other: Not classified

Conclusions:

According to CLP regulation, the test chemical cannot be classified for acute dermal toxicity, as the LD50 value is >2000 mg/kg bw.

Executive summary:

In different studies, the given 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 reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

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. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Hence, it was  concluded  that  the  LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were occlusively treated with the given test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

 

The above study is supported with another study conducted on rats for the test chemical. The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

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. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Therefore, it was  concluded  that the LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were occlusively treated with the given test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

 

Both the above studies are further supported with the study mentioned in publication and database. The acute dermal toxicity study of the given test chemical was conducted in rabbits at the dose concentration of 5000 mg/kg bw. No mortality was observed at 5000 mg/kg bw. Hence, LD50 value was considered to be >5000 mg/kg bw, when rabbits were treated with test chemical by dermal application.

 

Thus, based on the above summarised 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, the given test chemical cannot be classified for acute dermal toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Data is Klimisch 2 and from report.

Additional information

Acute oral toxicity:

In different studies, the given 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 for test chemical. The studies are summarized as below –

 

Acute oral toxicity study of the given test chemical was conducted in 10 rats at the dose concentration range of 100-1600 mg/kg bw. The given test chemical (undiluted) was administered via oral route. Animals were observed for mortality, body weight change and clinical signs for 14 days.

Mortality was observed within 2 hrs to 1 day in treated animals at 800 mg/kg bw. Symptoms included weakness, roughening of the coat, cyanosis, tremors, darkening of the eyes, and lacrimation just before death. Body weight change was observed in more than 6 animals.

Under the condition of this, the LD50 value was considered to be 800 mg/kg bw, when 10 rats were treated with the given test chemical via oral route.

 

The above study is supported with another study mentioned in study report for the test chemical. The reported study was designed and conducted to determine the acute oral toxicity profile of the given test chemical as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) in Sprague Dawley rats.

Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in ptosis, distension, reduced locomotor activity, ataxic gait and tremors with onset at 10 minutes to 2 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 (Step - II). Administration of the test item at 300 mg/kg resulted in ptosis, distension, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 4 hours after the dosing. One animal died on day 1 after the dosing. One 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 (Step - I). Administration of the test item at 2000 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing. Three animals died at 1 hour after the dosing.

All surviving animals from 300 mg/kg dose group exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in found dead as well as terminally sacrificed animals from 300 mg/kg and 2000 mg/kg dose groups.

Under the condition of this, the lethal concentration (LD50) value for acute oral toxicity test was considered in between 300-2000 mg/kg bw, when female Sprague Dawley rats were treated with the given test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

 

Both the above studies are further supported with the study mentioned in study report for the test chemical. The reported study was designed and conducted to determine the acute oral toxicity profile of the given test chemical as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) in Sprague Dawley rats.

Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait and tremors with onset at 30 minutes to 2 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 (Step - II). Administration of the test item at 300 mg/kg resulted in distension, piloerection, reduced locomotor activity, ataxic gait, tremors and loss of righting reflex with onset at 30 minutes to 4 hours after the dosing. One animal died at 6 hours after the dosing. One 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 (Step - I). Administration of the test item at 2000 mg/kg resulted in reduced locomotor activity, ataxic gait, tremors and convulsions with onset at 5 to 30 minutes after the dosing. Three animals died at 1 hour after the dosing.

All surviving animals from 300 mg/kg dose group exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in found dead animal and animals sacrificed terminally from 300 mg/kg dose group. Gross pathological examination revealed stomach distended with unabsorbed residual test item in found dead animals from 2000 mg/kg dose group.

Hence, it was concluded that the lethal concentration (LD50) value for acute oral toxicity test was considered in between 300-2000 mg/kg bw, when female Sprague Dawley rats were treated with the given test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is between 300-2000 mg/kg bw. Thus, comparing this range with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to the low vapour pressure of the test chemical, which is reported to be 0.01995164 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

Acute Dermal Toxicity:

In different studies, the given 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 reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

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. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Hence, it was  concluded  that  the  LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were occlusively treated with the given test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

 

The above study is supported with another study conducted on rats for the test chemical. The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

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. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Therefore, it was  concluded  that the LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were occlusively treated with the given test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

 

Both the above studies are further supported with the study mentioned in publication and database. The acute dermal toxicity study of the given test chemical was conducted in rabbits at the dose concentration of 5000 mg/kg bw. No mortality was observed at 5000 mg/kg bw. Hence, LD50 value was considered to be >5000 mg/kg bw, when rabbits were treated with test chemical by dermal application.

 

Thus, based on the above summarised 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, the given test chemical cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is between 300-2000 mg/kg bw, for acute oral toxicity and LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this range and value with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute oral toxicity and cannot be classified acute dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.