(E)-3-(benzyl(4-((4-nitrophenyl)diazenyl)phenyl)amino)propanenitrile

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25 August 1998 to 11 September 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

429-530-4

EC Name:

-

Cas Number:

96662-24-7

Molecular formula:

Hill formula: C22H19N5O2 CAS formula: C22H19N5O2

IUPAC Name:

3-[benzyl({4-[2-(4-nitrophenyl)diazen-1-yl]phenyl})amino]propanenitrile

Test material form:

solid: particulate/powder

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Species: New Zealand albino rabbit
Origin: Charles River Deutschland GmbH
Number of animals: 3
Body weight at start of the study: 3.1 -3.8 kg
Animal maintenance: in fully air-conditioned rooms in separate cages arranged in a battery
Room temperature: 20 ± 3eC
Relative humidity: 50 ± 20 %
Lighting time: 12 hours daily
Acclimatisation: 1 week under study conditions
Food: ssniff K-H (V2333), ad libitum and hay (approx. 15 g daily)
Water: water from automatic water dispensers, ad libitum
Animal identification: numbered ear tags

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

Dose levels: 100 mg per eye, applied neat.

Duration of treatment / exposure:

24 hours duration

Observation period (in vivo):

Initially 72 hours extended to 7 days

Number of animals or in vitro replicates:

3

Details on study design:

About 24 hours before the start of the study the test eyes of all animals were examined under UV light for corneal lesions after instillation of one drop of a 0.01 % fluorescein-sodium solution. Only animals without ocular abnormalities were used for the study.

100 mg T-9601 was administered once to the conjunctival sac of the left eye of three rabbits. The untreated eyes served in each case as a control.

24 hours after administration and at all other designated examination times at which the treated eyes still showed discharge or at which a corneal examination with fluorescein sodium solution took place, the treated eyes were washed out thoroughly with isotonic saline at approx. 37 °C.

The eyes were examined 1, 24, 48 and 72 hours after administration of the test substance. At 24 and 72 hours as well as after 7, 14 days, the eyes were also examined for corneal lesions under UV light after instillation of one drop of a 0.01 % fluorescein-sodium solution. Lesions in cornea, iris or conjunctivae were graded numerically. All other changes or toxic effects were recorded. Since effects were still present in the eyes after 72 hours, further examinations were carried out after 7 days.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

iris score

Basis:

mean

Remarks:

all animals

Time point:

24/48/72 h

Score:

0

Max. score:

2

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

animal #1

Time point:

24/48/72 h

Score:

1.67

Max. score:

3

Reversibility:

fully reversible within: 7 days

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

animal #2

Time point:

24/48/72 h

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Remarks:

animal #3

Time point:

24/48/72 h

Score:

0.33

Max. score:

3

Reversibility:

fully reversible within: 48 hours

Results 6

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animal #1

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 hours

Results 7

Irritation parameter:

chemosis score

Basis:

mean

Remarks:

animal #2 and #3

Time point:

other: overall 24, 48 & 72 hours

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

7 days after application the irritations observed were all reversible.

Other effects:

From one hour up to 3 days after application the conjunctiva of the animal showed definitely injected blood vessels up to deeper crimson red colours and slight swellings up to swelling with partial eversion of lids. The irritations were attended by clear coloured eye discharge.

Any other information on results incl. tables

Generalised Results By Animal – Animals Against Parameters – Multiple Times

Activity : - Eye Irritation Results

Study: 98.0575 – T-9601 – Eye Irritation Test in Rabbits

Group Sex	Animal Number	Day Number	Time Slot	Bodyweight g	Treated Eye	Control Eye normal	Duration Treatment	Initial Pain Reaction	Eye Irrigated	Fl.ceine staining	Opacity Grade	Opacity Area	Iris	Redness	Chemosis	Discharge
1f	57	1	Treatment	3480	Left	Yes	24 hour	None
			1 hour after treatment			Yes			No		0	0	0	1	2	2
		2	24 hours after treatment			Yes			Yes	0	0	0	0	2	2	1
		3	48 hours after treatment			Yes			No		0	0	0	2	1	0
		4	72 hours after treatment			Yes			Yes	0	0	0	0	1	0	0
		8	7 days after treatment			Yes			Yes	0	0	0	0	0	0	0
	607	1	Treatment	3180	Left	Yes	24 hour	None
			1 hour after treatment			Yes			No		0	0	0	2	2	2
		2	24 hours after treatment			Yes			Yes	0	0	0	0	2	0	0
		3	48 hours after treatment			Yes			No		0	0	0	0	0	0
		4	72 hours after treatment			Yes			Yes	0	0	0	0	0	0	0
	681	1	Treatment	3770	Left	Yes	24 hour	None
			1 hour after treatment			Yes			No		0	0	0	2	1	1
		2	24 hours after treatment			Yes			Yes	0	0	0	0	1	0	0
		3	48 hours after treatment			Yes			No		0	0	0	0	0	0
		4	72 hours after treatment			Yes			Yes	0	0	0	0	0	0	0

Nominal Dose: Group 1 – 0.1g

 

Clinical Observation – Clinical Signs by Animal

Study: 98.0575 – T-9601 – Eye Irritation Test in Rabbits

Day numbers relative to Start Date
Group Sex	Animal Number	Clinical Signs	1	2	3	4	8
			1 hour p. appl.	24 hours p. appl.	48 hours p. appl.	72 hours p. appl.	7 days p. appl.
1f	57	No Abnormalities Detected	X	-	X	X	X
		Eye discharge serous	M	S	-	-	-
	607	No Abnormalities Detected	-	X	X	X	-
		Eye discharge serious	M	-	-	-	-
	681	No Abnormalities Detected	-	X	X	X	-
		Eye discharge serious	S	-	-	-	-

X = Present  S = slight  M = Moderate

Nominal Dose: Group 1 – 0.1g

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Testing of T-9601 for primary eye irritation in the rabbit showed, that the substance is not irritating to eye.

Executive summary:

Testing for primary irritation in the rabbit eye provides information on ocular lesions after a single administration of the test substance to the conjunctival sac of the eye and serves as a basis for classification and labelling.

 

The present study was conducted in compliance with EEC-Guideline B.5."Acute Toxicity Eye Irritation" of the Directive 92/69/EEC: Commission Directive of July 31,1992 adapting to technical progress for the seventeenth time Council Directive 67/548/EEC on the approximation of the laws, regulations and administrative provisions relating to the classification, packag­ing and labelling of dangerous substances and OECD Guidelines for Testing of Chemicals, 405 „Acute Eye Irritation / Corrosion", updated guideline, adopted: February 24, 1987.This study was conducted in compliance with the Principles of Good Laboratory Practice (GLP).

 

From one hour up to 3 days after application the conjunctiva of the animal showed definitely injected blood vessels up to deeper crimson red colours and slight swellings up to swelling with partial eversion of lids. The irritations were attended by clear coloured eye discharge.

7 days after application the irritations were reversible.

 

Based on the individual scores after 24, 48 and 72 hours, the following mean values were calculated:

 

Opacity of cornea	All animals	0.00	Iris	All animals	0.00
	Animal 57	0.00		Animal 57	0.00
	Animal 607	0.00		Animal 607	0.00
	Animal 681	0.00		Animal 681	0.00
Redness of conjunctiva	All animals	0.89	Chemosis of conjunctiva	All animals	0.33
	Animal 57	1.67		Animal 67	1.00
	Animal 607	0.67		Animal 607	0.00
	Animal 681	0.33		Animal 681	0.00

 

Testing of T-9601 for primary eye irritation in the rabbit showed, that the substance is not irritating to eye.