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

Acute Toxicity: oral

Administrative data

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

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

EC Number:

240-474-8

EC Name:

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

Cas Number:

16423-68-0

Molecular formula:

C20H8I4O5.2Na

IUPAC Name:

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

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Erythrosine(FD & C Red No. 3)
- Molecular formula (if other than submission substance): C20-H6-I4-Na2-O5
- Molecular weight (if other than submission substance): 879.86
- InChl Key (if other than submission substance): RAGZEDHHTPQLAI-UHFFFAOYSA-L
-Substance type- Organic
- Physical state: Solid (powder)
-Purity: 90% pure colouring,
- Impurities (identity and concentrations): 10% lower iodinated fluorescein (c. 6%), sodium iodide (c. 2%), volatile matter (< 1%) and water (< 1%)

Test animals

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

Administration / exposure

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

Results and discussion

Effect levels

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

Any other information on results incl. tables

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

Applicant's summary and conclusion

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”.