1,4-Benzenediamine, N,N'-mixed Ph and tolyl derivs.

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: This is a guideline study performed according to GLP standards. There were only slight deviations from the guidelines, but these did not affect the valididy of the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Slight modifications from the guideline were e.g. more accurate description and scoring of corneal effects and additional examination of aqueous humour. The observation of these parameters however did not affect the validity of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1,4-Benzenediamine, N,N'-mixed Ph and tolyl derivs.

EC Number:

273-227-8

EC Name:

1,4-Benzenediamine, N,N'-mixed Ph and tolyl derivs.

Cas Number:

68953-84-4

Molecular formula:

C18-20H16-20N2

IUPAC Name:

Reaction mass of N-phenyl,N'-o-tolyl-phenylene diamine, N,N'-diphenyl-p-phenylene diamine and N,N'-di-o-tolyl-phenylene diamine

Test animals / tissue source

Species:

rabbit

Strain:

other: HC:NZW

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Adult albino rabbits were received from Interfauna UK, Ltd., Wyton, Huntington, England. Prior to the experiment experiments were checked to establish that they were biologically normal.
- Age at study initiation: adult, not further specified
- Weight at study initiation: 3.4-3.8 kg
- Housing: Rabbits were individually housed under standardized conventional conditions. Bedding was changed at least twice weekly.
- Diet (e.g. ad libitum): A standard diet of approx. 100 - 120g per animal per day was provided (Ssniff K 4, Ssniff Spezialdiäten GmbH, Soest). Organisms were fed once a day (morning).
- Water (e.g. ad libitum): Tap water (drink water quality) was provided at libitum.
- Acclimation period: animals were acclimatized for at least 14 days. Concurrently, animals were monitored for diseases and faeces was examined for Coccidia oocysts.

For identification purposes, rabbits were individually ear marked.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 + 3 °C
- Humidity (%): approximately 50%
- Air changes (per hr): air exchange rate was maintained at approx. 10 times per hour
- Photoperiod (hrs dark / hrs light): 12hr light/12hr dark photocycle. Irradiance was kept at approximately 27 Watt/m2

Prior to the initiation of the experiment, animals were weighed and randomly assigned to a treatment.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100 µl of the pulverized substance (equivalent to 45 mg)
- Concentration (if solution): not applicable

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

Three rabbits per administration route were used (in accordance with the German Animal Protection Act and the OECD guideline).

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The treated eye was rinsed with saline.
- Time after start of exposure: 24h


OBSERVATIONS

Recorded according to Draize et al.
- signs of cornea (opacity and area affected)
- iris (hyperaemia, reaction to light)
- conjunctivae (erythema, chemosis)
- discharge
Recorded according to McDonald and Shadduck
- aqueous humour (opacity)

Other serious lesions or toxic effects were also recorded.

Only effects which persisted for more than 24h were included in the evaluation. The irritation indices/mean irritation indices were calculated for cornea, iris, erythema and swelling of the conjunctivae. As 3 animals were used, interpretation was based on the individual indices obtained from the two most sensitive animals.

To define epithelial damage, one drop of 1% fluorescein solution was applied to the corneal surface 24h after administration of the test substance. Where positive results were observed, this was repeated at the later observation period. The eye was then rinsed with saline to remove excess and nonabsorbed fluorescein. Evaluation was performed by means of UV illumination (area) in a darkended room and diffuse white illumination (intensity) according to McDonald and Shadduck.

Where photographically possible, pictures of significant signs were taken after 72h after application and at the end of the observation period.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

other: all animals

Time point:

other: 24, 48 and 72h

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Results 2

Irritation parameter:

iris score

Basis:

other: all animals

Time point:

other: 24, 48 and 72 hr

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Results 3

Irritation parameter:

conjunctivae score

Basis:

animal: G15 and G13

Time point:

other: 24, 48 and 72 hr

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal: G11

Time point:

other: 24 hr

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 48 hours

Results 5

Irritation parameter:

conjunctivae score

Basis:

animal: G11

Time point:

other: 48 and 72 hr

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Results 6

Irritation parameter:

chemosis score

Basis:

other: all animals

Time point:

other: 24, 48 and72 hr

Score:

0

Max. score:

0

Reversibility:

other: not applicable

Any other information on results incl. tables

Animal no.	Sex, body weight	Tissue	Signs	Draize grades							Irrit.
				1h	24h	48h	72h	7d	14d	21d
G15	F; 3.8 kg	Cornea	o	0	0	0	0	0	-	-	0
			a	0	0	0	0	0	-	-
		Fluorescein	i	-	0	-	-	-	-	-
			a	-	0	-	-	-	-	-
		Iris		0	0	0	0	0	-	-	0
		Conjunctivae	r	1	0	0	0	0	-	-	0
			s	0	0	0	0	0	-	-	0
		Aqueous humour		0	0	0	0	0	-	-
		Discharge		2	1	0	0	0	-	-
G11	F; 3.4 kg	Cornea	o	0	0	0	0	0	-	-	0
			a	0	0	0	0	0
		Fluorescein	i	-	0	-	-	-	-	-
			a	-	0	-	-	-	-	-
		Iris		0	0	0	0	0	-	-	0
		Conjunctivae	r	1	1	0	0	0	-	-	0.3
			s	0	0	0	0	0	-	-	0
		Aqueous humour		0	0	0	0	0	-	-
		Discharge		1	1	0	0	0	-	-
G13	F; 3.7 kg	Cornea	o	0	0	0	0	0	-	-	0
			a	0	0	0	0	0	-	-
		Fluorescein	i	-	0	-	-	-	-	-
			a	-	0	-	-	-	-	-
		Iris		0	0	0	0	0	-	-	0
		Conjunctivae	r	1	0	0	0	0	-	-	0
			s	0	0	0	0	0	-	-	0
		Aqueous humour		0	0	0	0	0	-	-
		Discharge		1	0	0	0	0	-	-

o = opacity

s = area

i = intensity

r = redness

s = swelling

- = not examined

The test substance was not irritating, Slight conjunctivitis was observed 1h after application at all animals. By day 21 all animals were free of irritation signs.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Criteria used for interpretation of results: EU

Conclusions:

The substance was not found irritating to the eye (exposure period 24h). The mild and transient reactions of the mucous membranes immediately following exposure of the solid test substance into the eye are considered as mechanically induced effects.