Disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

Endpoint summary

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on different studies conducted on rats and mice for the test chemical. The LD50 value is >2000 mg/kg bw for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:

The study doesnot need to be conducted because exposure of humans via inhalation isnot likely taking in to account, the vapor pressure of the test chemical and/or the possibility of exposure to aerosols, dusts/mists and vapours is highly unlikely. Taking into account the low vapour pressure of the substance (7.572711605e-21 Pascal), acute toxicity by inhalation is unlikely.

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 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:

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 assess the Toxicological profile of test item to a single administration via oral route to Sprague Dawley rats. This study was designed to determine the acute toxicity at fixed dose levels by oral route of the test item.

GLP compliance:

yes

Test type:

acute toxic class 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: Female rats of the age of approximately 8 to 12 weeks old were used at the commencement of its dosing.
- Weight at study initiation: The weight were within ± 20% of the mean weight of any animal used for dosing. Body weight range was 197.8 to 205.9 grams.
Body weights at the start :
Female
Mean : 201.57 g (= 100 %)
Minimum : 197.8 g (- 1.87 %)
Maximum : 205.9 g (+ 2.15 %)
Total No. of animals : 12
- 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): Room temperature was maintained at 20.6 to 23.2 degree centigrade.
- Humidity (%): Room humidity was maintained at 54.2% to 58.6%.
- Air changes (per hr): The animal room was independently provided with at least 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: To: No data

Route of administration:

oral: gavage

Vehicle:

other: Distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: No data
- Amount of vehicle (if gavage): No data
- Justification for choice of vehicle: No data
- Lot/batch no. (if required): No data
- Purity: No data

MAXIMUM DOSE VOLUME APPLIED: The test item was administered in the dose volume of 10 ml/kg body weight.

DOSAGE PREPARATION (if unusual): No data
CLASS METHOD (if applicable) No data
- Rationale for the selection of the starting dose: No data

Doses:

Dose Group I - 300 mg/kg
Dose Group I - 300 mg/kg
Dose Group II - 2000 mg/kg
Dose Group II - 2000 mg/kg

No. of animals per sex per dose:

Three females were used at each step.

Control animals:

yes

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.

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.

Statistics:

No data

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:

Group I
Step I :
All animals survived through the study period of 14 days.

Group I
Step II :
All animals survived through the study period of 14 days .

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 diarrhea (reddish colour stools) in one animal with onset at 2 hours after the dosing. All animals survived through the study period of 14 days and were free of sign

Gross pathology:

Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.

Other findings:

No data

Table No. I - Summary of Clinical Signs of Toxicity and Mortality

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 (Reddish colour stools)	1	1	2 hrs. - 6 hrs.	0/3
		No clinical signs observed	2	2, 3	Day 0 to Day 14

 

 

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	2	4, 5	Day 0 to Day 14	0/3
		Diarrhoea (Reddish colour stools)	1	6	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	2000	Diarrhoea (Reddish colour stools)	1	7	30 min. - 6 hrs.	0/3
		No clinical signs observed	2	8, 9	Day 0 to Day 14

 

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	2	10, 12	Day 0 to Day 14	0/3
		Diarrhoea (Reddish colour stools)	1	11	2 hrs. - 6 hrs.

 

Staining of the stool is attributed to the dark red colour of the test item.

 

Table No.II

 

Mean Body Weight and Percent Body Weight Gain (g)

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	200.70	214.30	6.78	230.70	7.66	14.95
		± SD	1.25	1.90	1.37	1.85	0.95	0.76

 

 

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	201.10	214.80	6.81	232.40	8.19	15.56
		± SD	0.89	3.18	1.23	3.72	0.15	1.48

 

 

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	202.30	216.50	7.02	232.83	7.55	15.09
		± SD	1.01	3.11	1.06	2.36	0.59	0.59

 

 

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	202.17	215.80	6.75	230.87	6.99	14.21
		± SD	4.09	3.72	0.33	2.22	0.84	1.23

 

 

Table No.III

 

Summary of Gross Pathological Findings

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	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

Appendix No.I

 

Individual Animal - Clinical Signs of Toxicity and Mortality

Test System : Sprague Dawley Rat

Sex : Female

 Group I :   

 

Step No.	Dose mg/kg	Total Number of Animals	Observed Signs	Animal Nos.	Period of signs in days From - to	Mortality
I	300	3	Diarrhoea (Reddish colour stools)	1	2 hrs. - 6 hrs.	0
			No clinical signs observed	2	Day 0 - Day 14	0
			No clinical signs observed	3	Day 0 - Day 14	0

 

  Group I :

Step No.	Dose mg/kg	Total Number of Animals	Observed Signs	Animal Nos.	Period of signs in days From - to	Mortality
II	300	3	No clinical signs observed	4	Day 0 - Day 14	0
			No clinical signs observed	5	Day 0 - Day 14	0
			Diarrhoea (Reddish colour stools)	6	1 hr. - 6 hrs.	0

 

  Group II :

Step No.	Dose mg/kg	Total Number of Animals	Observed Signs	Animal Nos.	Period of signs in days From - to	Mortality
I	2000	3	Diarrhoea (Reddish colour stools)	7	30 min. - 6 hrs.	0
			No clinical signs observed	8	Day 0 - Day 14	0
			No clinical signs observed	9	Day 0 - Day 14	0

 

  Group II :

Step No.	Dose mg/kg	Total Number of Animals	Observed Signs	Animal Nos.	Period of signs in days From - to	Mortality
II	2000	3	No clinical signs observed	10	Day 0 - Day 14	0
			Diarrhoea (Reddish colour stools)	11	2 hrs. - 6 hrs.	0
			No clinical signs observed	12	Day 0 - Day 14	0

 

Staining of the stool is attributed to the dark red colour of the test item.

 

 

 

 

Appendix No.II

 

Individual Animal - Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex : Female

Group : I                       Step I :                                 Dose  : 300 mg/kg body weight

Animal No.	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
1	201.9	212.4	5.20	230.8	8.66	14.31
2	199.4	214.3	7.47	228.8	6.77	14.74
3	200.8	216.2	7.67	232.5	7.54	15.79

 

Group : I                       Step II :                                 Dose  : 300 mg/kg body weight

Animal No.	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
4	201.8	215.3	6.69	232.8	8.13	15.36
5	200.1	211.4	5.65	228.5	8.09	14.19
6	201.4	217.7	8.09	235.9	8.36	17.13

 

Group : II                      Step I :                                 Dose  : 2000 mg/kg body weight

Animal No.	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
7	202.1	217.2	7.47	232.6	7.09	15.09
8	203.4	219.2	7.77	235.3	7.34	15.68
9	201.4	213.1	5.81	230.6	8.21	14.50

 

Group : II                      Step II :                                 Dose  : 2000 mg/kg body weight

Animal No.	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
10	205.9	219.3	6.51	232.7	6.11	13.02
11	202.8	216.2	6.61	231.5	7.08	14.15
12	197.8	211.9	7.13	228.4	7.79	15.47

 

 

Appendix No.III

 

Individual Animal - Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex : Female

Group : I

Step I :

Dose  : 300 mg/kg body weight

Animal No.	Fate	Gross Pathological Findings
1	TS	No abnormality detected
2	TS	No abnormality detected
3	TS	No abnormality detected

 

Group : I

Step II :

Dose  : 300 mg/kg body weight

Animal No.	Fate	Gross Pathological Findings
4	TS	No abnormality detected
5	TS	No abnormality detected
6	TS	No abnormality detected

 

Group : II

Step I :

Dose  : 2000 mg/kg body weight

Animal No.	Fate	Gross Pathological Findings
7	TS	No abnormality detected
8	TS	No abnormality detected
9	TS	No abnormality detected

 

Group : II

Step II :

Dose  : 2000 mg/kg body weight

Animal No.	Fate	Gross Pathological Findings
10	TS	No abnormality detected
11	TS	No abnormality detected
12	TS	No abnormality detected

 

TS = Terminal sacrifice

Interpretation of results:

other: not classified

Conclusions:

It was concluded that the acute oral median lethal dose (LD50) of test chemical, when administered to Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus,according to the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute oral toxicant. CLP Classification: “Not classified”.

Executive summary:

The study now reported was designed and conducted to determine the acute oral toxicity profile of test chemical 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 (reddish colour stools) in one animal with onset at 2 hours and no 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 (Step - II). Administration of the test item at 300 mg/kg resulted in diarrhoea (reddish colour stools) in one animal with onset at 1 hour and no mortality after the dosing.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 (Step - I).Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools) in one animal with onset at 30 minutes and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools) in one animal with onset at 2 hours and no mortality after the dosing. All animals from 300 mg/kg and 2000 mg/kg dose groups survived through the study period of 14 days.Staining of the stool is attributed to the dark red colour of the test item.Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.It was concluded that the acute oral median lethal dose (LD₅₀) of test chemical, when administered to Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus,according to the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute oral toxicant. CLP Classification: “Not classified”.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

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

Data waiving:

exposure considerations

Justification for data waiving:

the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size

Endpoint conclusion

Endpoint conclusion:

no study available

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:

male/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: [no]
- Age at study initiation: Young adult male and female rats aged between 6 – 9 weeks were used.
- Weight at study initiation: The weight range of approximately 217.7 to 254.5 grams at initiation of dosing were used.
Body weights at the start :
Male
Mean : 246.00 g (= 100 %)
Minimum : 235.8 g (- 4.15 %)
Maximum : 254.5 g (+ 3.46 %)
Total No. of animals : 5
Female
Mean : 223.30 g (= 100 %)
Minimum : 217.7 g (- 2.51 %)
Maximum : 227.9 g (+ 2.06 %)
Total No. of animals : 5
- Fasting period before study: No data
- 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): Room temperature was maintained at 20.0 to 22.3 degree centigrade.
- Humidity (%): Room humidity was maintained at 55.7% to 59.6%.
- Air changes (per hr): The animal room was independently provided with at least 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: No data

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

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: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): 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 bw
- Constant volume or concentration used: yes
- For solids, paste formed: No data

VEHICLE
- Amount(s) applied (volume or weight with unit):No data
- Concentration (if solution): No data
- Lot/batch no. (if required): No data
- Purity: No data

Duration of exposure:

24 hours

Doses:

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).

No. of animals per sex per dose:

10 (5/sex).

Control animals:

yes

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 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).

Statistics:

No data

Preliminary study:

no data available

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

Mortality:

Sex : Male
Group I -
All animals survived through the study period of 14 days.

Sex : Female
Group I -
All animals survived through the study period of 14 days.

Clinical signs:

other: 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 a

Gross pathology:

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

Other findings:

- 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.

Table No. I - Summary of Clinical Signs of Toxicity and Mortality

 Test System : 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	0 - 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	0 - 14	0/5

 

 

Table No. II

 

Summary of Evaluation of Dermal Reaction

 

Test System : 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	0 - 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	0 - 14	0/5

 

 

 

Table No.III

 

Mean Body Weight and Percent Body Weight Gain (g)

 

Test System : 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	246.00	270.58	10.00	287.98	6.34	17.02
		± SD	7.71	8.29	1.46	19.65	4.54	6.12

 

 

 

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	223.30	237.00	6.14	248.78	4.96	11.41
		± SD	4.39	5.47	1.62	7.04	1.04	2.07

Table No.IV

 

Summary of Gross Pathological Findings

 Test System : 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

Appendix No.I

 

Individual Animal - Clinical Signs of Toxicity and Mortality

 

Test System : 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	0 - 14	0
				2	0 - 14	0
				3	0 - 14	0
				4	0 - 14	0
				5	0 - 14	0

 

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	0 - 14	0
				7	0 - 14	0
				8	0 - 14	0
				9	0 - 14	0
				10	0 - 14	0

 

Appendix No.II

 

Individual Animal - Evaluation of Dermal Reaction

 

Test System : Sprague Dawley Rat

Sex : Male  

Group : I

Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
1	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
2	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
3	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
4	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
5	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

 

 

Sex : Female  

Group : I

Dose  : 2000 mg/kg body weight

Animal	Dermal	D A Y S
No.	Reaction	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14
6	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
7	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
8	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
9	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
10	Erythema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0
	Oedema	0	0	0	0	0	0	0	0	0	0	0	0	0	0	0

Appendix No.III

 

Individual Animal - Body Weight and Percent Body Weight Gain (g)

 

Test System : Sprague Dawley Rat

Sex    : Male

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	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
1	235.8	264.2	12.04	284.4	7.65	20.61
2	241.2	260.4	7.96	256.0	-1.69	6.14
3	246.3	271.5	10.23	292.5	7.73	18.76
4	252.2	276.5	9.64	301.7	9.11	19.63
5	254.5	280.3	10.14	305.3	8.92	19.96

 

Sex    : Female

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	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
6	217.7	235.5	8.18	247.1	4.93	13.50
7	220.4	230.6	4.63	238.3	3.34	8.12
8	223.2	234.7	5.15	248.5	5.88	11.34
9	227.3	239.0	5.15	253.0	5.86	11.31
10	227.9	245.2	7.59	257.0	4.81	12.77

 

Appendix No.IV

 

Individual Animal - Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/007

Test System : Sprague Dawley Rat

Sex    : Male

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	Fate	Gross Pathological Findings
1	TS	No abnormality detected
2	TS	No abnormality detected
3	TS	No abnormality detected
4	TS	No abnormality detected
5	TS	No abnormality detected

 

 

Sex    : Female

Group : I

Dose  : 2000 mg/kg body weight

 

Animal No.	Fate	Gross Pathological Findings
6	TS	No abnormality detected
7	TS	No abnormality detected
8	TS	No abnormality detected
9	TS	No abnormality detected
10	TS	No abnormality detected

 

TS = Terminal sacrific

Interpretation of results:

other: Not classified

Conclusions:

It was concluded that the acute dermal median lethal dose (LD50) of test chemical, when administered to male and female Sprague Dawley rats was considered to be greater than 2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibits acute toxicity by the dermal route.CLP Classification: “Not classified”.

Executive summary:

The study now reported was designed and conducted to determine the acute dermal toxicity profile of test chemical 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. It was concluded that the acute dermal median lethal dose (LD₅₀) of test chemical, when administered to male and female Sprague Dawley rats was considered to be greater than 2000 mg/kg body weight. Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibits acute toxicity by the dermal route. CLP Classification: “Not classified”.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

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

The study reported 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 Guidelines. 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 (reddish colour stools) in one animal with onset at 2 hours and no 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 (Step - II). Administration of the test item at 300 mg/kg resulted in diarrhoea (reddish colour stools) in one animal with onset at 1 hour and no mortality after the dosing. 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 (Step - I).Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools) in one animal with onset at 30 minutes and no mortality after the dosing. As no mortality were observed at 24 hours after the dosing, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in diarrhoea (reddish colour stools) in one animal with onset at 2 hours and no mortality after the dosing. All animals from 300 mg/kg and 2000 mg/kg dose groups survived through the study period of 14 days. Staining of the stool is attributed to the dark red colour of the test item. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups. It was concluded that the acute oral median lethal dose (LD50) of test chemical, when administered to Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to the CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

 

The above study is supported with another study performed on rats by using test chemical at dose range of Male:5800-9400 mg/ kg and Female: 6100-7700 mg/ kg. The rats were maintained at 21±1OC and humidity at 50-60% for 7 days. Rats was given Spillers’ Laboratory Small Animal Diet and tap-water ad lib. Aqueous solutions of EBS were administered in single doses to groups of five males and five females by oral intubation. The animals were observed for 7 days after treatment and autopsies were carried out on animals that died during this period and on randomly-selected survivors at day 7. Acute LD50values with 95% confidence limits were calculated by the method of Weil. Abdomen was distended and the faeces were colored deep red whereas the urine was only slightly colored. The small intestines and caeca were found to be distended and filled with a pink fluid. The colon frequently contained a compact, solid mass and faecal production was reduced. The LD50 value of test chemical on rats was considered to be 7400 mg/ kg (95% CI:5800-9400) for male rats and 6800 mg/ kg (95% CI:6100-7700) for female rats when treated with test chemical orally via gavage.

 

Both the above studies are further supported by the acute oral toxicity test conducted on Charles river strained female rat which were administered with test chemical by oral route.10 animals per dose were dosed with 0,2000,2600,3380 and 4394 mg/kg .At highest dose 4394 mg/kg all animals were died on day 1 and 2 and at dose of 3380 mg/kg 7 animals were died out of 10 hence, LD50 value was considered to be 2891 ± 1.02 mg/kg in acute oral toxicity of test chemical administered to female rat by oral route.

Moreover, Acute oral toxicity study was performed in rats using test chemical at dose concentration of 7100 mg/kg bw.50% mortality was observed at dose 7100 mg/kg bw. Hence, LD50 value was considered to be 7100 mg/kg bw,when rats were treated with test chemical orally.

Furthermore, Acute oral toxicity test of test chemical was carried out with 4 male ddY mice. Test substance was administered orally to group of four male mice at 2000 mg/kg dose. The acute oral LD50 value of test chemical is considered to be >2000 mg/kg in male ddY mice.

Again the above studies were supported by another oral toxicity test conducted on male mice when administered with test chemical by oral route.10 animals per dose were dosed with 0(water at 40 ml/kg),1000,2000,4000 and 8000 mg/kg. The animals were observed each day for 5 consecutive days and the number of dead per dose per day was recorded. All control animals exhibited no observable effects after oral administration of water. At highest dose 4913 mg/kg 100% mortality was observed and at dose 2890 mg/kg 6 animals were died out of 10. Hence,LD50 value was considered to be 2558 ± 1.35 mg/kg in acute oral toxicity of test chemical administered to male mice by oral route.

The above studies were furthermore supported by the acute toxicity test performed on mouse by using test chemical at range of doses for Male: 5800-7800 mg/kg and female: 4900-9800 mg/kg. Mouse was maintained in a room at 21±1°C with a relative humidity of 50-60% and was given Spillers’ Laboratory Small Animal Diet and tap-water ad lib. Aqueous solutions of test chemical were administered in single doses to groups of five males and five females by oral intubation. The animals were observed for 7 days after treatment and autopsies were carried out on animals that died during this period and on randomly-selected survivors at day 7. Acute LD50 values with 95% confidence limits were calculated by the method of Weil. After oral dosage there was a general lethargy, followed either by recovery or by coma and death. In many instances the abdomen was distended. The colon frequently contained a compact, solid mass and faecal production was reduced. The faeces were coloured deep red whereas the urine was only slightly coloured. Therefore, the LD 50 value for the test chemical on mouse was considered to be for male 6700 mg/kg bw (95% CI:5800 -7800 mg/kg bw) and for female 6900 mg/kg bw (95% CI:4900-9800 mg/kg bw) when test chemical was administered orally via gavage.

All the above studies contradict with the study conducted on rats using test chemical at dose concentration of 1895 mg/kg bw.50% mortality was observed at dose 1895 mg/kg bw. Hence, the LD50 value was considered to be 1895 mg/kg bw, when rats were treated with test chemical orally.

Again the above study is supported with contradictory study performed on Osborne-Mendel rats for 3 weeks. Test chemical was administered orally via gavage to groups of 5 male and 5 female. 230-275 g for the males and from 190 to 235 g for the females were used for study. The surviving rats were observed for approximately 3 wk after treatment. The surviving rats were observed for approximately 3 week after treatment. Clinical signs like depression, diarrhoea with dye in the faeces, and pink ears, tail, paws and nose were observed. In rats that died shortly after administration of the compound, the internal organs, muscle and fat were also pink. Rats killed at the end of the observation period still had dye-stained fur in the anal region and dye-stained tails, but organs and fat were normal in appearance.The oral LD50 of test chemical was calculated to be 1840 mg/kg with 95 % confidence limits of 1614-2098 mg/kg; the slope function was 1.36 with 95% confidence limits of 1.1-1.7.Hence,LD50 value was considered to be 1840 mg/kg (95% CI:1614-2098),when rats were treated with test chemical orally.

All the above studies were again supported by another contradictory study in mice using test chemical at dose concentration of 1264 mg/kg bw.50% mortality was observed at dose 1264 mg/kg bw. Hence, LD50 value was considered to be 1264 mg/kg bw,when 10 male Albino mice were treated with test chemical orally.

Based on the available results, it is evident that the test chemical is likely to be non-toxic when dosed orally to rats and mice with LD50 value of greater than 2000 mg/kg. But some contradictory results in rats as well as mice suggest a possibility of the test chemical to cause toxic response when dosed orally with LD50 value between 1000 -2000 mg/kg. Negating these contradictory and considering the results from the OECD 423 Guideline study report as well as other reliable sources, the acute oral LD50 for the test chemical can be concluded to be > 2000 mg/kg and classified under the category “Not Classified” as per CLP Regulation.

Acute Inhalation Toxicity:

The study doesnot need to be conducted because exposure of humans via inhalation is not likely taking in to account, the vapor pressure of the test chemical and/or the possibility of exposure to aerosols, dusts/mists and vapours is highly unlikely. Taking into account the low vapour pressure of the substance (7.572711605e-21 Pascal), acute toxicity by inhalation is unlikely.

Acute Dermal toxicity:

The study reported was designed and conducted to determine the acute dermal toxicity profile of test chemical 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. It was concluded that the acute dermal median lethal dose (LD50) of test chemical, when administered to male and female Sprague Dawley rats was considered to be greater than 2000 mg/kg body weight. Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibits acute toxicity by the dermal route. CLP Classification: “Not classified”. Thus, based on the above experimental study 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 was >2000 mg/kg bw for acute oral and acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the test chemical can be classified under the category "NOT CLASSIFIED".