Disodium 4,4'-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2'-disulphonate

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 4,4'-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2'-disulphonate

EC Number:

240-245-2

EC Name:

Disodium 4,4'-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2'-disulphonate

Cas Number:

16090-02-1

Molecular formula:

C40H38N12Na2O8S2

IUPAC Name:

disodium 2,2'-ethene-1,2-diylbis{5-[(4-anilino-6-morpholin-4-yl-1,3,5-triazin-2-yl)amino]benzenesulfonate}

Test material form:

other: fine granules

Details on test material:

Identification : Disodium 4,4’-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2’-disulphonate
Appearance : Free flowing Fine Granules
Manufactured date: July, 2014
Expiry Date : June, 2015
Storage conditions : Room temperature (20 - 30 °C)

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Species : Rabbit (Oryctolagus cuniculus)
Strain : New Zealand White
Age : 3.5 to 4.7 Months (Approximately)
Sex : Male
Number of Animals : Three
Supplier/Source : Procured from GENTOX BIOSERVICES PVT. LTD.
Health Status : Healthy young adult rabbits were used for the study
Body weight of animals : Minimum: 2.090 kg & Maximum: 2.354 kg (Prior to Treatment)
Acclimatisation :Rabbits were acclimatised to the test conditions for a period of 6 days (Animal No. 1) and 9 days (Animal No. 2 and 3) prior to the application of the test item.
Identification : During acclimatization marking was done with non toxic marker pen in the inside of left ear of rabbits and after acclimatization, animals were marked with permanent number in the inner side of right ear of rabbits. Permanent marker and cage card was used for identification. The individual cage cards were labelled with study no., study type, test system, sex, dose, animal number, experimental start and completion date.
Husbandry Conditions
Diet : All animals were provided conventional laboratory rabbit diet
Water : Aqua guard filtered tap water was provided ad libitum.
Husbandry : The animals were housed individually in stainless steel 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 minimum twice a week.
Experimental Room Condition
Temperature : Minimum: 18.70 °C Maximum: 21.60 °C
Relative humidity : Minimum: 49.10% Maximum: 69.40%
Light-dark-rhythm : 12 hour light and 12 hour dark
Air Changes : More than 12 changes per hour

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

Preparation of Application Site
Approximately 24 h prior to treatment, the fur coat of each rabbit was removed from dorsal lumbar region approximately 6 X 6 cm at contralateral sites on each rabbit using clipper, one as control and other site as treatment. Rabbits with healthy intact skin were selected for the study.

Test Item Application Procedure
The 0.5 ml of test item (as such) was applied uniformly over clipped area (approximately 6 X 6 cm) of the trunk of each rabbit skin (treated site) and 0.5 ml distilled water was applied at control site. Test item was held in contact with the skin with a porous gauze dressing and non-irritating tape (Micropore 3”) throughout a 4-hour exposure period, to prevent access by the rabbits to the patch and resultant ingestion of the test item. At the end of the exposure period, residual test item was removed by using cotton soaked in distilled water.
A single rabbit (Animal No. 1) was used for initial testing. The patch was removed after 4 h and the responses graded, no erythema and oedema was observed at 1 hour, 24 hour, 48 hour and 72 hour in animal no. 1. Hence the confirmatory test was conducted on additional two rabbits (No. 2 and 3) after 24 hour to confirm the non irritant nature of the test item.

Duration of treatment / exposure:

4 hours

Observation period:

72 hours

Number of animals:

3

Details on study design:

A dose of 0.5 gm of test item moistened with 0.5 ml of distilled water was applied to the skin, over an area of approximately 6 x 6 cm clipped of hair on one side of rabbits.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 24 hrs

Score:

0.67

Max. score:

0

Reversibility:

fully reversible

Results 2

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 24 hrs

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 3

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 48 hrs

Score:

0.33

Max. score:

0

Reversibility:

fully reversible

Results 4

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 48 hrs

Score:

0

Max. score:

0

Reversibility:

fully reversible

Results 5

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: 72 hrs

Score:

0

Max. score:

0.67

Reversibility:

fully reversible

Results 6

Irritation parameter:

edema score

Basis:

mean

Time point:

other: 72 hrs

Score:

0

Max. score:

0

Reversibility:

fully reversible

Irritant / corrosive response data:

No erythema and edema (skin irritation) were found at the end of 72 hour observation period after patch removal.

Other effects:

no effects observed

Any other information on results incl. tables

Table 1

Skin Reaction

 

In Treated area Dose:0.5 g (Pulverized form)                                                     Sex:Male

 

Animal No.	Test	Treated  area	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Right	1	1	1	0	0	0	0	0
2	Confirmatory	Left	1	1	0	0	0	0	0	0
3		Right	1	1	1	0	0	0	0	0

 

 

 

 

In Control areaDose:0.5 ml of distilled water                                                   Sex:Male

 

Animal No.	Test	Treated area	Erythema score				Oedema score
			1h	24h	48h	72h	1h	24h	48h	72h
1	Initial	Left	0	0	0	0	0	0	0	0
2	Confirmatory	Right	0	0	0	0	0	0	0	0
3		Left	0	0	0	0	0	0	0	0

Key: h = Hour.

Erythema                                                                                                       Oedema

0 =No erythema                                                                                           0 =No oedema

1 = Very slight erythema(barely perceptible)

Table 1 Continued…

Mean Individual Animal Score at 24, 48 and 72 hours (Treated Site)

 

Animal Number                   Observations	1	2	3
Erythema	0.67	0.33	0.67
Oedema	0.00	0.00	0.00

Table 2

Individual Animal BodyWeight

Sex:Male

Animal No.	Body Weight (kilogram)
	Prior to Dosing	At termination
1	2.090	2.178
2	2.144	2.172
3	2.354	2.386

Table 3

Individual AnimalClinicalSigns

Sex:Male

Animal No.	Days (Post dosing Observation)
	0	1	2	3
1	1	1	1	1
2	1	1	1	1
3	1	1	1	1

Key: 1 = Normal.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

No erythema and no oedema (skin irritation) were found at the end of 72 hour observation period after patch removal.
Hence, it was concluded that Disodium 4,4’-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2’-disulphonate (CAS No. – 16090-02-1) was Non-Irritating to the skin of Male New Zealand White rabbits under the experimental conditions tested and
Not Classified as Skin Irritant as per CLP regulation.

Executive summary:

In acute dermal irritation/corrosion Study of Disodium 4,4’-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2’-disulphonate (CAS No. – 16090-02-1)in Rabbits,  No erythema and no oedema (skin irritation) were found at the end of 72 hour observation period after patch removal.

Hence, it was concluded that Disodium 4,4’-bis[(4-anilino-6-morpholino-1,3,5-triazin-2-yl)amino]stilbene-2,2’-disulphonate (CAS No. – 16090-02-1)was Non-Irritating to the skin of Male New Zealand White rabbitsunder the experimental conditions tested and not Classified as Skin Irritant as per CLP regulation.