Benzenamine, 3,3'-[[1,1'-biphenyl]-4,4'-diylbis(oxy)]bis-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted in compliance with OECD Guideline for Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion". Adopted 24 April 2002. There was no deviation from the protocol.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

yes (incl. QA statement)

Remarks:

The Department of Health of the Government of the United Kingdom

Test material

Test material information

Constituent 1

Reference substance name:

3-{[4'-(3-aminophenoxy)-[1,1'-biphenyl]-4-yl]oxy}aniline

EC Number:

700-102-1

Cas Number:

105112-76-3

Molecular formula:

C24H20N2O2

IUPAC Name:

3-{[4'-(3-aminophenoxy)-[1,1'-biphenyl]-4-yl]oxy}aniline

Details on test material:

- Name of test material (as cited in study report): 3,3-(4,4-biphenylenedioxy)dianiline
- Substance type: Raw material for polymer
- Physical state: Off-white powder
- Analytical purity: >99 %
- Lot/batch No.: 200709501
- Expiration date of the lot/batch: End April 2008
- Storage condition of test material: Room temperature
- Other: Date of receipt was 28 August 2007

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Stock supply
- Age at study initiation: at least 39 weeks
- Weight at study initiation: 4.08 to 4.79 kg
- Housing: Each animal was housed individually in a plastic cage with perforated floors at the Eye Research Centre, Eye, Suffolk.
- Diet (e.g. ad libitum): Each animal was offered 125 g of a standard laboratory rabbit diet (Rabbit GD Diet Pellets supplied by Highgate Farm) per day
- Water (e.g. ad libitum): Drinking water was supplied ad libitum
- Acclimation period: at least 17 weeks


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 to 20
- Humidity (%): 40 to 70
- Air changes (per hr): not available
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL or 80 mg

Duration of treatment / exposure:

Single ocular dose

Observation period (in vivo):

96 hours

Number of animals or in vitro replicates:

Three

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done):
- Time after start of exposure:


SCORING SYSTEM:
Criteria for assessment of pain response (not a component of the regulatory guidelines):
No response (class 0): No initial pain;
A few blinks only; normal within one or two minutes (class 1): Practically no initial pain;
Rabbit blinks and tries to open eye but the reflexes close it (class 2): Slight initial pain;
Rabbit holds eye shut and puts pressure on lids; may rub eye with paw, may vocalise (class 3): Moderate initial pain;
Rabbit holds eye shut vigorously, may tilt head, may have reluctance to rub eye; may vocalise (class 4): Severe initial pain;
Rabbit holds eye shut very firmly; may vocalise, claws at eye, jumps and tries to escape (class 5): Very severe initial pain

Criteria for assessment of pain response (not a component of the regulatory guidelines):
No response (class 0): No initial pain;
A few blinks only; normal within one or two minutes (class 1): Practically no initial pain;
Rabbit blinks and tries to open eye but the reflexes close it (class 2): Slight initial pain;
Rabbit holds eye shut and puts pressure on lids; may rub eye with paw, may vocalise (class 3): Moderate initial pain;
Rabbit holds eye shut vigorously, may tilt head, may have reluctance to rub eye; may vocalise (class 4): Severe initial pain;
Rabbit holds eye shut very firmly; may vocalise, claws at eye, jumps and tries to escape (class 5): Very severe initial pain

The untreated eye was used as a comparison with the treated eye during assessment of ocular lesions.
Ocular irritation was assesses using the prescribed numerical system:

Cornea Opacity: degree of density (area most dense taken for reading)
No opacity = 0;
Scattered or diffuse area of opacity (other than slight dulling of normal lustre), details of iris clearly visible = 1;
Easily discernible translucent areas, details of iris slightly obscured = 2;
Nacreous areas, no details of iris visible, size of pupil barely discernible = 3;
Opaque cornea, iris not discernible through the opacity = 4

Area of cornea involved:
None = 0;
One quarter (or less) but not zero = 1;
Greater than one quarter, but less than half = 2;
Greater than half, but less than three quarters = 3;
Greater than three quarters, up to whole area = 4

The presence or absence of corneal ulceration or stippling was designated positive or negative.

Iris:
Normal = 0;
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia or injection, any of these or combination of any thereof, iris still reacting to light (sluggish reaction is positive) = 1;
No reaction to light, haemorrhage, gross destruction (any or all of these) = 2

Conjunctivae Redness: (refers to the most severe reading of palpebral and bulbar conjunctivae, as compared to the control eye)
Blood vessels normal = 0;
Some blood vessels definitely hyperaemic (injected) = 1;
Diffuse, crimson colour, individual vessels not easily discernible = 2;
Diffuse beefy red = 3

Chemosis: (lids and/or nictating membranes)
No swelling = 0;
Any swelling above normal (includes nictating membranes) = 1;
Obvious swelling with partial eversion of lids = 2;
Swelling with lids about half-closed = 3;
Swelling with lids more than half-closed =4

The presence or absence of ulceration or necrosis was designated positive or negative.

Discharge:
No discharge = 0;
Any amount different from normal = 1;
Discharge with moistening of lids and hairs adjacent to lids = 2;
Discharge with moistening of lids and hairs and considerable area around eye = 3;
Extensive mucoid discharge = 4

An ophthalmoscope or pencil beam torch was available for use to facilitate inspection of the eyes

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 1 hour after instillation

Score:

2

Reversibility:

fully reversible

Results 2

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

other: 24 to 72 hours

Score:

0

Reversibility:

fully reversible

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 1 hour after instillation

Score:

2

Reversibility:

fully reversible

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

other: 24 to 72 hours

Score:

0

Reversibility:

fully reversible

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: 1 hour after instillation

Score:

2

Reversibility:

fully reversible

Results 6

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: 24 hours after instillation

Score:

2

Reversibility:

fully reversible

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

other: 48 to 72 hours

Score:

0

Reversibility:

fully reversible

Irritant / corrosive response data:

Injection of the conjunctival blood vessels was apparent in each treated eye one hour after instillation persisting in one case 23 hours later. The treated eye of each animal was overtly normal 48 hours after instillation.
Instillation of the test material gave rise to practically no or slight initial pain response.

Any other information on results incl. tables

Table 1: Mean values for ocular lesions, Mean irritation scores

	Time after instillation of test substance
Area of eye	1 hour	24 hours	48 hours	72 hours
Cornea	0.0	0.0	0.0	0.0
Iris	0.0	0.0	0.0	0.0
Conjunctiva	2.0	0.7	0.0	0.0
Total mean score	2.0	0.7	0.0	0.0

Table 2: Grades for ocular irritation responses following instillation of the test substance

Animal number and sex: 49F*		Pain evaluation response: 0
Region of the eye	Response	Grade of response at time after instillation after hours
		1	24	48	72
Cornea	Opacity (A)	0	0	0	0
	Area (B)	0	0	0	0
	Ulceration	-	-	-	-
	Stippling	-	-	-	-
Corneal Score (A x B x 5)		0	0	0	0
Iris	Value (C)	0	0	0	0
Iridial Score (C x 5)		0	0	0	0
Conjunctiva	Redness (D)	1	0	0	0
	Chemosis (E)	0	0	0	0
	Discharge (F)	0	0	0	0
	Necrosis	-	-	-	-
	Ulceration	-	-	-	-
Conjunctival Score ((D+E+F)x2)		2	0	0	0

* Sentinel animal

F: Female

Table 2: continued

Animal number and sex: 81F		Pain evaluation response: 0
Region of the eye	Response	Grade of response at time after instillation after hours
		1	24	48	72
Cornea	Opacity (A)	0	0	0	0
	Area (B)	0	0	0	0
	Ulceration	-	-	-	-
	Stippling	-	-	-	-
Corneal Score (A x B x 5)		0	0	0	0
Iris	Value (C)	0	0	0	0
Iridial Score (C x 5)		0	0	0	0
Conjunctiva	Redness (D)	1	0	0	0
	Chemosis (E)	0	0	0	0
	Discharge (F)	0	0	0	0
	Necrosis	-	-	-	-
	Ulceration	-	-	-	-
Conjunctival Score ((D+E+F)x2)		2	0	0	0

F: Female

Table 2: continued

Animal number and sex: 82F		Pain evaluation response: 0
Region of the eye	Response	Grade of response at time after instillation after hours
		1	24	48	72
Cornea	Opacity (A)	0	0	0	0
	Area (B)	0	0	0	0
	Ulceration	-	-	-	-
	Stippling	-	-	-	-
Corneal Score (A x B x 5)		0	0	0	0
Iris	Value (C)	0	0	0	0
Iridial Score (C x 5)		0	0	0	0
Conjunctiva	Redness (D)	1	1	0	0
	Chemosis (E)	0	0	0	0
	Discharge (F)	0	0	0	0
	Necrosis	-	-	-	-
	Ulceration	-	-	-	-
Conjunctival Score ((D+E+F)x2)		2	2	0	0

F: Female

Applicant's summary and conclusion

Interpretation of results:

other: "practically non-irritating"

Remarks:

Criteria used for interpretation of results: other: Kay and Calandra 1962

Conclusions:

The highest total mean score was 2.0 occurring at the 1 hour observation; accordingly under the criteria Kay and Calandra (1962) 3,3-(4,4-biphenylenedioxy)dianiline was classified as "practically non-irritating" to the eye. 3,3-(4,4-biphenylenedioxy)dianiline did not require labelling with the risk phrase R36, "Irritating to eyes", in accordance with Commission Directive 2001/59/EC and was assigned to US EPA Category 4.

Executive summary:

A study was performed to assess the eye irritation potential of 3,3-(4,4-biphenylenedioxy)dianiline to the rabbit. The study was carried out in compliance with OECD Guideline for Testing of Chemicals No. 405 "Acute Eye Irritation/Corrosion".

Three rabbits were each administered a single ocular dose of a volume of 0.1 mL of the test substance weighing approximately 80 mg and observed for four days after instillation. Injection of the conjunctival blood vessels was apparent in each treated eye one hour after instillation persisting in one case for 23 hours. The treated eye of each animal was overtly normal 48 hours after instillation. Instillation of the test substance gave rise to no or practically no pain response. The highest total mean score was 2.0 occurring at the 1 hour observation; accordingly under the criteria of Kay and Calandra (1962), the test substance was classified as "practically non-irritating" to the eye. The substance 3,3'-[biphenyl-4,4'-diylbis(oxy)]dianiline did not require labelling with the risk phrase R36, "Irritating to eyes", in accordance with Commission Directive 2001/59/EC and was assigned to US EPA Category 4.