Disodium 2,4-diamino-3-{[4-(ethenylsulfonyl)phenyl]diazenyl}-5-{[4-(ethenylsulfonyl)-2-sulfonatophenyl]diazenyl}benzenesulfonate

Endpoint summary

Description of key information

The LD50 value for FAT 40866 in rats via oral route was found to be greater than 2000 mg/kg bw

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

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to OECD guideline 423 and in accordance with GLP

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Guidelines followed

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Environment: with temperature 21 to 23°C, relative humidity 65 to 67%, with 12 hours light and 12 hours dark cycle.

Animals were housed individually in standard polysulfone cages
(Size: approximately L 425 x B 266 x H 175 mm), with stainless steel top grill having facilities for pelleted food and drinking water in polycarbonate bottle.

Route of administration:

oral: gavage

Vehicle:

other: Milli-Q water

Details on oral exposure:

VEHICLE, Milli-Q water was used as vehicle to prepare the test item suspension.

MAXIMUM DOSE VOLUME ADMINISTERED: 10 mL/kg body weight

DOSAGE PREPARATION (if unusual):

The test item formulation was prepared on each day of the test item administration.
G1 - [2000 mg/kg body weight - Treatment (FTS and STS)] : A quantity of 5.53# (5.0x1.106) g of test item was weighed in a mortar and mixed by adding small volume of Milli-Q water and stirred by using a pestle. The content was then transferred to a measuring cylinder. The mortar was rinsed again with Milli-Q water and the rinsing was transferred to the measuring cylinder. The mortar was rinsed with vehicle and the rinsing was then transferred to the measuring cylinder. The volume was made up to 25 mL to get the test item concentration of 200 mg/mL suspension.

Homogeneity of the test item in the vehicle was maintained by constant stirring using glass rod. Preparations were made prior (within 1 hour) to dosing.
The preparation corresponding only to the first step of each dose was sent for analysis.

#: While calculating the required weight of test item for preparation of dose formulation, purity correction factor of 1.106 was applied. While calculating the concentration of the prepared dose formulation, a correction factor of 1/1.106 was applied, which was calculated as below:
Purity correction = 100/90.4 = 1.106

Doses:

2000 mg/kg

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:day 1, day 7 and day 14.
- Necropsy of survivors performed: yes

The rats surviving to the end of the observation period were euthanised using isoflurane anaesthesia and subjected to detailed necropsy by an experienced prosector and the findings were recorded in raw data. Microscopic examination was not carried out as no gross pathological changes were observed.

- Clinical Signs and Pre-Terminal Deaths :
The animals were observed five times on test day 1 (day of administration) i.e. at 30 minutes and four times at hourly intervals and once daily during days 2 to 15 post administration.

Observations included changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems and somatomotor activity and behaviour pattern. Attention was directed to the observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma and findings of all observations were recorded.

-Body Weights
The body weights were recorded on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration).

-Necropsy
The rats surviving to the end of the observation period were euthanised using isoflurane anaesthesia and subjected to detailed necropsy by an experienced prosector and no abnormalities were observed at Nectopsy.

Statistics:

None

Preliminary study:

None

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: Non toxic

Mortality:

No data

Clinical signs:

G1 - [2000 mg/kg body weight - Treatment (FTS and STS)]: There were no clinical signs were observed, however reddish brown faeces was observed in all rats on day 2, the change in colour is due to the colour of the test item. All the rats were normal from day 3 onwards. There were no pre-terminal deaths.

Body weight:

G1 - [2000 mg/kg body weight - Treatment (FTS and STS)]: The body weights of all the rats increased throughout the observation period.

Gross pathology:

No abnormality was detected at necropsy.

Other findings:

None

TABLE 1.      Body weight, Body weight change and Pre-terminal deaths

Group and Dose (mg/kg body weight)	Rat No.	Sex	Body weight (g)					No. dead/  No. tested
			Initial (Day 1)	8^thday	Weight change (day 8 – Initial)	15^thday	Weight change           (day 15 – Initial)
G1 (FTS) 2000	Rm2811	F	208.3	213.0	4.7	220.4	12.1	0/3
	Rm2812	F	195.6	208.1	12.5	217.8	22.2
	Rm2813	F	205.7	215.2	9.5	223.2	17.5
G1 (STS) 2000	Rm2814	F	210.2	215.1	4.9	221.8	11.6	0/3
	Rm2815	F	212.3	216.8	4.5	223.1	10.8
	Rm2816	F	195.7	200.3	4.6	207.6	11.9

 F: Female;                   FTS: First Treatment Step;                STS: Second Treatment Step

APPENDIX 1.      Individual Toxic Signs and Necropsy Findings

Group: G1 (FTS)                   Dose: 2000 mg/kg body weight

Rat No.

Sex

Total volume admin (mL)

Day of Observation

Necropsy Findings

Day 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

30 min

1hour

2 hours

3 hours

4 hours

Rm2811

F

2.1

N

N

N

N

N

RBF

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm2812

F

2.0

N

N

N

N

N

RBF

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm2813

F

2.1

N

N

N

N

N

RBF

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female;               FTS: First Treatment Step;                N: Normal;              admin: administered;                       min: minutes;            

mL: millilitre            NAD: No Abnormality Detected     RBF : Reddish Brown Faeces

                       

Group: G1 (STS)                  Dose: 2000 mg/kg body weight

Rat No.

Sex

Total volume admin (mL)

Day of Observation

Necropsy Findings

Day 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

30 min

1hour

2 hours

3 hours

4 hours

Rm2814

F

2.1

N

N

N

N

N

N^@

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm2815

F

2.1

N

N

N

N

N

N^@

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

Rm2816

F

2.0

N

N

N

N

N

N^@

N

N

N

N

N

N

N

N

N

N

N

N

N

NAD

F: Female;                 STS: Second treatment step;                N: Normal;                admin: administered;           min: minutes; 

mL: millilitre             NAD: No Abnormality Detected         @: Reddish Brown Faeces

Interpretation of results:

practically nontoxic

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

Based on the results of the present study, the test item, FAT 40866/A TE is classified as follows:
-“Category 5” or Unclassified as per Globally Harmonized Classification system of Annex 2d of the Guideline, OECD 423.
- The LD50 is 5000 mg/kg body weight or Unclassified as per LD50 cut-off value.

Executive summary:

The study was performed to access the acute oral toxicity of test item, FAT 40866/A TE in Wistar rats.  

The test item mixed in Milli-Q water was administered as oral gavage to overnight fasted (16 to 18 hours) 3 female rats at the dose of 2000 mg/kg body weight (G1-FTS). There were no clinical signs of toxicity and pre-terminal deaths. Based on the scheme - Annex 2d of the guideline OECD 423, three additional female rats were tested at the same dose of 2000 mg/kg body weight (G1-STS). There were no clinical signs of toxicity and pre-terminal deaths. 

All rats gained weight during experimental period. The rats were subjected to necropsy at termination and there were no abnormalities detected at necropsy.

Based on the results of the present study, the test item, FAT 40866/A TE is classified as follows:

“Category 5” or Unclassified as per Globally Harmonized Classification system of Annex 2d of the Guideline, OECD 423.

 The LD50 is 5000 mg/kg body weight or Unclassified as per LD50 cut-off value.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

The study was performed to access the acute oral toxicity of test item, FAT 40866/A TE in female Wistar rats.  

The test item mixed was administered as oral gavage at the dose of 2000 mg/kg body weight.There were no clinical signs of toxicity and pre-terminal deaths. All rats gained weight during experimental period. The rats were subjected to necropsy at termination and there were no abnormalities detected at necropsy.

Based on the results of the present study, the test item, FAT 40866/A TE is classified as follows:

“Category 5” or Unclassified as per Globally Harmonized Classification system of Annex 2d of the Guideline, OECD 423.

The LD50 is >2000 mg/kg bw or Unclassified as per LD50 cut-off value.

Justification for selection of acute toxicity – oral endpoint
The study was conducted according to OECD guideline 423 and in accordance with GLP

Justification for classification or non-classification

Based on the results of the present study, the test item, FAT 40866/A TE is classified as follows:

“Category 5” or Unclassified as per Globally Harmonized Classification system of Annex 2d of the Guideline, OECD 423.

 The LD50 is greater than 2000 mg/kg bw or Unclassified as per LD50 cut-off value.