1-dodecylpyridinium chloride

Endpoint summary

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on experimental study conducted on rats for the test chemical.The study concluded that the LD50 value isbetween50-300 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 3” 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 0.0046 Pa. 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 experimental study conducted on rats for the test chemical. The study concluded that the LD50 value is >1000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified in “Category 4 (>1000-2000 mg/kg bw)” 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 study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Principles of method if other than guideline:

The purpose of this study was to access the toxicological profile of the test item to a single administration via oral route to Sprague-Dawley rats.

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

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 192.2 to 207.1 grams.
Body weights at the start : Female Mean: 199.74 g (= 100 %); Minimum : 192.2 g (- 3.78 %); Maximum : 207.1 g (+ 3.68 %)
- 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): 20.0 to 22.0 degree centigrade.
- Humidity (%): 54.4% to 59.3%
- 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:13-10-2017 to 31-10-2017

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle:300 mg/kg, 50 mg/kg and 50 mg/kg
MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight.
DOSAGE PREPARATION (if unusual): The test item was dissolved in distilled water. The formulation was prepared fresh. The test item was administered in the dose volume of 10 ml/kg bw.

Doses:

Dose Group I : 300 mg/kg
Dose Group II : 50 mg/kg
Dose Group II : 50 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: 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.

Statistics:

not specified

Preliminary study:

not specified

Sex:

female

Dose descriptor:

LD50

Effect level:

> 50 - 300 mg/kg bw

Based on:

test mat.

Mortality:

Group I Step I : Two animals died at 6 hours and one animal died on day 1 after the dosing.
Group II Step I : All animals survived through the study period of 14 days.
Group II Step II : All animals survived through the study period of 14 days.

Clinical signs:

other: Group I Step I : Animals treated at the dose level of 300 mg/kg body weight resulted in diarrhoea and reduced locomotor activity with onset at 30 minutes to 1 hour after the dosing. Group II Step I : Animals treated at the dose level of 50 mg/kg body wei

Gross pathology:

Gross pathological examination did not reveal any abnormalities in all animals from 300 mg/kg and 50 mg/kg dose groups.

Other findings:

not specified

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

 

Laboratory Test Item Code :TAS/122/054

Test System : 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	Diarrhoea	3	1 2 3	1 hr. - 4 hrs. 30 min. - 4 hrs. 1 hr. - 6 hrs.	3/3
		Reduced locomotor activity	3	1 2 3	1 hr. - 4 hrs. 30 min. - 4 hrs. 1 hr. - 6 hrs.

 

Group II :

Step No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days From - to	Mortality
I	50	Diarrhoea	3	4,5 6	1 hr. - 4 hrs. 1 hr. - 6 hrs.	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	50	Diarrhoea	3	7 8,9	1 hr. - 6 hrs. 1 hr. - 4 hrs.	0/3

 

Table No.II 

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/054

Test System : 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	195.23	-	-	-	-	-
		± SD	2.87	-	-	-	-	-

 

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	50	Mean	201.77	214.87	6.49	232.20	8.07	15.09
		± SD	3.40	4.15	0.30	5.14	1.78	1.98

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	50	Mean	202.23	215.20	6.41	232.37	7.97	14.89
		± SD	4.54	5.58	0.58	6.80	0.93	0.85

 

Table No.III

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/054

Test System : Sprague Dawley Rat

Sex : Female  Group I :

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	300	1 - 3	FD	No abnormality detected

Group II : 

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
I	50	4 - 6	TS	No abnormality detected

 

Group II : 

Step No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	50	7 - 9	TS	No abnormality detected

 

                    FD = Found dead

         TS = Terminal sacrifice 

Interpretation of results:

Category 3 based on GHS criteria

Conclusions:

Under the condition of the study, the acute oral LD50 value of the given test chemical was considered in between 50 – 300 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category 3” according to criteria of CLP.

Executive summary:

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 diarrhoea and reduced locomotor activity with onset at 30 minutes to 1 hour after the dosing. Two animals died at 6 hours and one animal died on day 1 after the dosing.

As mortality was observed at 300 mg/kg dose group, additional three female animals were treated with the lower dose of 50 mg/kg of the test item (Step - I).

Administration of the test item at 50 mg/kg resulted in diarrhoea with onset at 1 hour after the dosing and no mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three additional female animals were added to the study and treated with the dose of 50 mg/kg of the test item (Step - II). Administration of the test item at 50 mg/kg resulted in diarrhoea with onset at 1 hour after the dosing and no mortality at 24 hours after the dosing.

All animals from 50 mg/kg dose group survived through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in all animals from 300 mg/kg and 50 mg/kg dose groups.

Under the condition of the study, the acute oral LD50 value of the given test chemical was considered in between 50 – 300 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category 3” according to criteria of CLP.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

200 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from study report.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

other justification

Justification for data waiving:

other:

Clinical signs:

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:

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 study report.

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Principles of method if other than guideline:

This study was designed to determine the dermal LD50 of the test item (up to 2000 mg/kg) or to establish a non-lethal dose level of 2000 milligram of test item per kilogram of body weight.

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: Young adult female rats aged between 8 – 10 weeks were used.
- Weight at study initiation: The weight ranges of approximately 222.8 to 245.7 grams at initiation of dosing were used.
Body weights at the start : Female Mean: 235.02 g (= 100 %); Minimum : 222.8 g (- 5.20 %); Maximum : 245.7 g (+ 4.54 %)
-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): 19.2 to 21.8 degree centigrade.
- Humidity (%): 56.0% to 59.1%
- 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: 11-09-2017 to 28-12-2017

Type of coverage:

occlusive

Vehicle:

water

Remarks:

(Distilled water)

Details on dermal exposure:

TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the total 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

Duration of exposure:

24 hours

Doses:

Dose Range Finding Study:
Group I : 200 mg/kg body weight
Group I : 1000 mg/kg body weight
Group I : 2000 mg/kg body weight
Main Study:
Group II : 1000 mg/kg body weight

No. of animals per sex per dose:

Total No. of animals : 5

Control animals:

not specified

Details on study design:

- 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 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:

Dose Range Finding Study: In the dose range finding study a single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered with the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight revealed no clinical signs of toxicity or mortality during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 1 000 mg/kg bw

Based on:

test mat.

Mortality:

Dose Range Finding Study:
Group I : Animal treated at the dose level of 200 mg/kg body weight survived through the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight survived through the study period of 14 days.
Group I : Animal treated at the dose level of 2000 mg/kg body weight found dead on day 1.
Main Study:
Group II : Animals treated at the dose level of 1000 mg/kg body weight survived through the study period of 14 days.

Clinical signs:

other: Dose Range Finding Study: Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days. Group I : Animal treated at the dose level of 1000 mg/kg body weight did not result

Gross pathology:

Gross pathological examination did not reveal any abnormalities attributable to the treatment in animals from 200 mg/kg dose group whereas gross pathological examination revealed moderate to severe erythema on site of application in animals from 1000 mg/kg dose group from dose range finding study and main study sacrificed terminally.

Other findings:

Evaluation of Dermal Reaction
Dose Range Finding Study:
Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight resulted in minimal to moderate erythema at the site of application from day 1 to day 14.
Main Study:
Group II : Animal treated at the dose level of 1000 mg/kg body weight resulted in minimal to moderate erythema at the site of application from day 1 to day 14.

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

Laboratory Test Item Code :TAS/122/054

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal No.	Period of signs in days  From - to	Mortality
I	200	No clinical signs observed	1	1	Day 0 - Day 14	0/1

 

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal No.	Period of signs in days  From - to	Mortality
I	1000	No clinical signs observed	1	2	Day 0 - Day 14	0/1

 

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal No.	Period of signs in days  From - to	Mortality
I	2000	Found dead	1	3	Day 1	1/1

 

Main Study:

Group  No.	Dose mg/kg	Observed Signs	Total Number of Animals	Animal Nos.	Period of signs in days  From - to	Mortality
II	1000	No clinical signs observed	2	4, 5	Day 0 - Day 14	0/2

Table No. II

Summary of Evaluation of Dermal Reaction

Laboratory Test Item Code :TAS/122/054

Test System : Sprague Dawley Rat

Sex : Female

DoseFinding Study:

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal No.	Period of signs in days  From - to
I	200	No dermal reaction observed	1	1	Day 0 - Day 14

 

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal No.	Period of signs in days  From - to
I	1000	Minimal to moderate erythema	1	2	Day 1 - Day 14

 

Main Study:

Group  No.	Dose mg/kg	Dermal Reaction	Total Number of Animals	Animal Nos.	Period of signs in days  From - to
II	1000	Minimal to moderate erythema	2	4, 5	Day 1 – Day 14

 

Table No.III

Mean Body Weight and Percent Body Weight Gain (g)

Laboratory Test Item Code :TAS/122/054

Test System : Sprague Dawley Rat

Sex : Female

DoseFinding Study:

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	200	Mean	226.0	241.6	6.90	252.6	4.55	11.77
		± SD	-	-	-	-	-	-

 

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	1000	Mean	235.1	248.8	5.83	257.4	3.46	9.49
		± SD	-	-	-	-	-	-

 

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	222.8	-	-	-	-	-
		± SD	-	-	-	-	-	-

 

Main Study:

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
II	1000	Mean	245.60	255.65	4.09	264.85	3.60	7.84
		± SD	0.14	1.34	0.61	0.78	0.24	0.38

Table No.IV

Summary of Gross Pathological Findings

Laboratory Test Item Code :TAS/122/054

Test System : Sprague Dawley Rat

Sex : Female

DoseFinding Study:

Group No.	Dose mg/kg	Animal Number	Animal Fate	Gross Pathological Findings
I	200	1	TS	No abnormality detected

 

Group No.	Dose mg/kg	Animal Number	Animal Fate	Gross Pathological Findings
I	1000	2	TS	Moderate to severe erythema was observed on site of application

 

Group No.	Dose mg/kg	Animal Number	Animal Fate	Gross Pathological Findings
I	2000	3	FD	No abnormality detected

 

                    Main Study:

Group No.	Dose mg/kg	Animal Numbers	Animal Fate	Gross Pathological Findings
II	1000	4, 5	TS	Moderate to severe erythema was observed on site of application

 

                     FD = Found dead

         TS = Terminal sacrifice

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >1000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical can be classified as an acute dermal toxicant. CLP Classification: “Category 4 (>1000-2000 mg/kg bw)”.

Executive summary:

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.

In the dose range finding study a single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered with the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight revealed no clinical signs of toxicity or mortality during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight.

Administration of 2000 mg/kg body weight resulted in mortality on day 1. As the dose range finding study revealed mortality at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals at a dose of 1000 mg/kg body weight. The animals were administered with a dose of 1000 mg/kg body weight in sequential manner at 48 hours intervals.

Administration of 200 mg/kg body weight exhibited normal body weight gain and no clinical signs of toxicity during the study period of 14 days. Administration of 1000 mg/kg body weight revealed minimal to moderate erythema at the site of application from day 1 to day 14 and exhibited normal body weight gain, no clinical signs of toxicity or mortality during the study period of 14 days from dose range finding study and main study. Gross pathological examination did not reveal any abnormalities attributable to the treatment in animals from 200 mg/kg dose group whereas gross pathological examination revealed moderate to severe erythema on site of application in animals from 1000 mg/kg dose group.

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >1000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical can be classified as an acute dermal toxicant. CLP Classification: “Category 4 (>1000-2000 mg/kg bw)”.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

1 000 mg/kg bw

Quality of whole database:

Data is Klimisch 1 and from study report.

Additional information

Acute oral toxicity:

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 diarrhoea and reduced locomotor activity with onset at 30 minutes to 1 hour after the dosing. Two animals died at 6 hours and one animal died on day 1 after the dosing. As mortality was observed at 300 mg/kg dose group, additional three female animals were treated with the lower dose of 50 mg/kg of the test item (Step - I).

Administration of the test item at 50 mg/kg resulted in diarrhoea with onset at 1 hour after the dosing and no mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three additional female animals were added to the study and treated with the dose of 50 mg/kg of the test item (Step - II). Administration of the test item at 50 mg/kg resulted in diarrhoea with onset at 1 hour after the dosing and no mortality at 24 hours after the dosing.

All animals from 50 mg/kg dose group survived through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in all animals from 300 mg/kg and 50 mg/kg dose groups.

Under the condition of the study, all rats were died at 300 mg/kg bw, hence the LD100 was considered to be 300 mg/kg bw. Considering this value, the LD50 value can be expected to be 200 mg/kg bw. Therefore, comparing this value / range with the criteria of CLP regulation, the given test chemical can be classified in “Category 3 (50 - ≤ 300)” 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 0.0046 Pa. 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 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.

In the dose range finding study a single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered with the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight revealed no clinical signs of toxicity or mortality during first 48 hours; hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight.

Administration of 2000 mg/kg body weight resulted in mortality on day 1. As the dose range finding study revealed mortality at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals at a dose of 1000 mg/kg body weight. The animals were administered with a dose of 1000 mg/kg body weight in sequential manner at 48 hours intervals.

Administration of 200 mg/kg body weight exhibited normal body weight gain and no clinical signs of toxicity during the study period of 14 days. Administration of 1000 mg/kg body weight revealed minimal to moderate erythema at the site of application from day 1 to day 14 and exhibited normal body weight gain, no clinical signs of toxicity or mortality during the study period of 14 days from dose range finding study and main study. Gross pathological examination did not reveal any abnormalities attributable to the treatment in animals from 200 mg/kg dose group whereas gross pathological examination revealed moderate to severe erythema on site of application in animals from 1000 mg/kg dose group.

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >1000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical can be classified as an acute dermal toxicant. CLP Classification: “Category 4 (1000-2000 mg/kg bw)”.

 

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that the LD50 value is between 50-300 mg/kg bw, for acute oral toxicity and LD50 value is >1000 mg/kg bw(1000-2000 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 3” for acute oral toxicity and “Category 4 (>1000-2000 mg/kg bw)” for acute dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.