Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)

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 report

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

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:

Acute oral toxicity study was performed to determine the toxicity potential of the test chemical after single administration in rats and an observation period of 14 days

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)

EC Number:

255-217-5

EC Name:

Hexasodium 2,2'-[vinylenebis[(3-sulphonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino]]bis(benzene-1,4-disulphonate)

Cas Number:

41098-56-0

Molecular formula:

C40H44N12O18S6.6Na

IUPAC Name:

hexasodium 2,2'-(ethene-1,2-diylbis{(3-sulfonato-4,1-phenylene)imino[6-(diethylamino)-1,3,5-triazine-4,2-diyl]imino})dibenzene-1,4-disulfonate

Test material form:

liquid

Details on test material:

- Name of the test chemical: Hexasodium 2,2'-(vinylenebis((3-sulphonato-4,1-phenylene)imino(6-(diethylamino)-1,3,5-triazine-4,2-diyl)imino))bis(benzene-1,4-disulphonate)
- Molceular formula: C40H38N12O18S6.6Na
- Molecular weight: 1305.1422 g/mol
- Smiles: CCN(CC)c1nc(nc(n1)Nc2cc(ccc2S(=O)(=O)[O-])S(=O)(=O)[O-])Nc3ccc(c(c3)S(=O)(=O)[O-])/C=C/c4ccc(cc4S(=O)(=O)[O-])Nc5nc(nc(n5)N(CC)CC)Nc6cc(ccc6S(=O)(=O)[O-])S(=O)(=O)[O-].[Na+].[Na+].[Na+].[Na+].[Na+].[Na+]

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS:
Species : Rattus norvegicus (Rat)
Strain :W istar
Number and Sex: Six Females
Supplier / Source: In-house animals, bred at Animal House, sa-FORD.CPCSEA Registration No. 1256/bc/09/CPCSEA.
Health Status: Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
Body weight of animals: Minimum: 149 g Maximum: 161 g (Individual body weights were within ± 20 % prior to treatment after overnight fasting)
Age : 8- 11 weeks at the time of dosing.
Acclimatisation: Animal nos. 1-3 were acclimatized for 6 days and 4-6 for 10 days, prior to administration of the test item.
Identification : The animals were marked temporarily on tail, permanently on toe pad micro tattooing and cage cards. Individual cage cards were labelled with study no., study type, test system, group, dose, sex, animal number experimental start and completion date.

Diet : All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No.: 400010 and 400011.
Bedding : All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) SPAR – 27 /2014.
Water : Aqua guard filtered tap water was provided ad libitum via drinking bottles.
Husbandry :T he animals were housed individually in polycarbonate cages.
Room Sanitation: The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
Cages and water bottle: All the cages and water bottles were changed at least twice every week

ENVIRONMENTAL CONDITIONS:
Temperature : Minimum: 19.40 °C Maximum: 22.10 °C
Relative humidity: Minimum: 46.60% Maximum: 65.40%
Light-dark-rhythm : 12 hour light and 12 hour dark
Air Changes : More than 12 changes per hour

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

Distilled water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 2000 mg/kg body weight
- Amount of vehicle (if gavage): No data
- Justification for choice of vehicle: based on solubility testing
- Lot/batch no.: No data

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg body weight

DOSAGE PREPARATION: The test chemical in desired quantity was added slowly in the vehicle and mixed well. Transferred the formulation to the measuring cylinder and made the final volume up to desired quantity of 10 ml. The dosing solution was prepared freshly, shortly prior to dose administration

Doses:

G1 = 2000 mg/kg body weight

No. of animals per sex per dose:

Six Female rats

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily
- Necropsy of survivors performed: yes, at the end of 14 day observation period, all the survived rats were euthanised by overdose of CO2 for external and internal observations.
- Other examinations performed:
Clinical Observation - After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all the surviving animals were observed once a day during the 14 day observation period.
Body weight - All surviving rats were weighed on days 0 (prior to dosing), 7 and 14. Animals were weighed immediately after found dead.
Mortality - All surviving animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.

Statistics:

Not specified

Results and discussion

Preliminary study:

Not specified

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: No mortality was observed

Mortality:

No mortality was observed in the animals treated with 2000 mg/kg dose throught out the 14 days observation period

Clinical signs:

other: At 2000 mg/kg, all the animals were normal throughout the experimental period

Gross pathology:

No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice

Other findings:

Not specified

Any other information on results incl. tables

 TABLES

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

Sex:Female

Animal No.	Group/ Dose (mg/kg)	Body Weight (gram)				Body Weight Change (%)
		Dose Volume*	Day 0	Day 7	Day 14	Day 0-7	Day 0-14
1	G1/ 2000	1.5	149	172	183	15.44	22.82
2		1.6	160	191	197	19.38	23.13
3		1.6	158	186	198	17.72	25.32
4		1.6	160	179	188	11.88	17.50
5		1.6	160	195	196	21.88	22.50
6		1.6	161	192	205	19.25	27.33

*= Dose volume calculated based on day 0 body weight.

 

 

Table 2: Summary of Animal Body Weight (g) and Body Weight Changes (%)

 

Sex:Female

Group/ Dose (mg/kg)		Rats Body Weight (g)			Body Weight Changes (%)
		Day 0	Day 7	Day 14	0-7	0-14
G1/ 2000	Mean	158.00	185.83	194.50	17.59	23.10
	SD	4.52	8.80	7.82	3.51	3.30
	n	6	6	6	6	6

Keys:SD = Standard Deviation, n = Number of Animals

Table 3: Individual Animal Clinical Signs and Symptoms

 

Sex:Female

Animal No.	Group/ Dose (mg/kg)	Hours (Day 0)
		1/2	1	2	3	4
1	G1/ 2000	1	1	1	1	1
2		1	1	1	1	1
3		1	1	1	1	1
4		1	1	1	1	1
5		1	1	1	1	1
6		1	1	1	1	1

 

Animal No.		Group/ Dose (mg/kg)		Days post dosing
			1		2		3		4		5		6		7		8		9		10		11		12		13		14
1		G1/ 2000		1		1		1		1		1		1		1		1		1		1		1		1		1		1
2			1		1		1		1		1		1		1		1		1		1		1		1		1		1
3			1		1		1		1		1		1		1		1		1		1		1		1		1		1
4			1		1		1		1		1		1		1		1		1		1		1		1		1		1
5			1		1		1		1		1		1		1		1		1		1		1		1		1		1
6			1		1		1		1		1		1		1		1		1		1		1		1		1		1

Table 4: Individual Animal Mortality Record

 

Sex:Female

Animal No.	Group/ Dose (mg/kg)	Day of Observation (Day 0 to 14)
		Morning Observations	Evening Observations
1	G1/ 2000	No mortality and morbidity	No mortality and morbidity
2		No mortality and morbidity	No mortality and morbidity
3		No mortality and morbidity	No mortality and morbidity
4		No mortality and morbidity	No mortality and morbidity
5		No mortality and morbidity	No mortality and morbidity
6		No mortality and morbidity	No mortality and morbidity

Table 5: Gross Necropsy Observation

 

Sex:Female                                                                                                                                        

Animal No.	Group/ Dose (mg/kg)	Mode of Death	Gross Observation
			External	Internal
1	G1/ 2000	TS	No abnormality detected	No abnormality detected
2		TS	No abnormality detected	No abnormality detected
3		TS	No abnormality detected	No abnormality detected
4		TS	No abnormality detected	No abnormality detected
5		TS	No abnormality detected	No abnormality detected
6		TS	No abnormality detected	No abnormality detected

Keys:TS= Terminal Sacrifice

Applicant's summary and conclusion

Interpretation of results:

other: Not Classified

Conclusions:

The acute oral LD50 value of the test chemical was >2000 mg/kg body weight. Thus cosidering the CLP criteria of classification, the test chemical was classified under the category "Not Classified".

Executive summary:

Acute Oral Toxicity Study of the test chemical was performed using female Wistar rats, This study was performed as per OECD No. 423. Six female Wistar rats were selected for acute oral toxicity study. The animals were fasted for minimum 16-18 hours prior to dosing and for 4 hours post dosing, food was withheld but drinking water provided ad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs. Three rats of first group were dosed with starting dose of 2000 mg/kg body weight and the animals did not show any mortality so another three animals of the same group were dosed with 2000 mg/kg body weight and no mortality was observed. Hence, further dosing was stopped. Body weights were recorded on day 0 (prior to dosing) 7 and 14. Mean body weight of all the animals treated with 2000 mg/kg body weight was observed with gain on day 7 and 14, as compared to day 0. At 2000 mg/kg, all the animals were normal throughout the experimental period. No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice. The acute oral LD₅₀ value of the test chemical was >2000 mg/kg body weight. Thus cosidering the CLP criteria of classification, the test chemical was classified under the category "Not Classified".