3,5-xylidine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 JAN 1990 - 09 JAN 1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Reference 2

Reference Type:

secondary source

Title:

Xylidine, BUA Stoffbericht 161

Year:

1994

Bibliographic source:

BUA, Beratergremium für umweltrelevante Altstoffe (BUA) der Gesellschaft Deutscher Chemiker, S.Hirzel Verlag 1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Principles of method if other than guideline:

Slight modifications from the guidelines (e.g: more accurate description and scoring of corneal defects, additional examination of aqueous humour) this does not affect the validity of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3,5-xylidine

EC Number:

203-607-0

EC Name:

3,5-xylidine

Cas Number:

108-69-0

Molecular formula:

C8H11N

IUPAC Name:

3,5-dimethylaniline

Details on test material:

Purity 98%

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Interfauna U.K. Ltd. Wyton, Huntingdon, England
- Age at study initiation: adult
- Weight at study initiation: 3.4 - 3.7 kg
- Housing: the rabbits were individually housed in stainless steel cages with flat rod bases or polyethylene cages with perforated bases, under standardized conventional conditions. Excrement trays beneath the cages contained low dust (wood) bedding (type S 8/15, Ssniff spezialdiaten GmbH, Soest). Bedding was regularly spot checked for contaminants at the instance of the Department of Laboratory Animal Services and changed at least twice weekly.
All the animals in this study were kept in one room. For capacity reasons, rabbits treated with other test substances were also housed in the same room. Adequate separation, clear cage and individual marks, as well as appropriate organization of working procedures ensured that the test animals were not mixed up.
- Diet (e.g. ad libitum): Standard diet Ssniff K 4, approx. 100 - 120g per animal/day; once per day in the morning. The nutritive composition and contaminant content of the standard diet were routinely spot checked and analysed at the instance of the department of laboratory animal services.
- Water (e.g. ad libitum): Tap water; ad libitum. The tap water was of drink water quality (Drink water ordinance of May 22, 1986, Federal Law Gazette (Bundesgesetzblatt) part I, page 760). The drink water was regularly examined at the instance of the Quality Assurance, Bayer AG. The results of the feed and water analyses provided no evidence of interference with the study objective. Drink water was supplied either in polycarbonate bottle containing approx. 750mL or from automatic watering.
- Acclimation period: Prior to the initiation of the treatment the animals were kept for at least 14 days in the quarantine station of the Dpartment of the Laboratory Animal Services and monitored for diseases. During this period pooled faeces specimens were examined for Coccidia oocysts.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3 °C
- Humidity (%): approx 50 %
- Air changes (per hr): approx 10 times per hour
- Photoperiod (hrs dark / hrs light): 12 hours, artificial illumination from 6 to 18hrs CET
The humidity and air temperature were continuously recorded using a calibrated thermohygrograph (type 252, Lambrecht Co. Gottingen). Occasional excursions from these conditions, e.g., due to cleaning of the room, had no detectable influence on the outcome of the study.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): After gently pulling the lower lid away from the eyeball a volume of 100 μL test substance was instilled into the conjunctival sac of one eye of each of three rabbits. The other eye remained untreated and served as control.

Duration of treatment / exposure:

24 hours

Details on study design:

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

SCORING SYSTEM & TOOL USED TO ASSESS SCORE:
Eye irritation was scored and recorded after administration given 1, 24, 48, 72h and 7 days. The signs of cornea (opacity and area affected), iris (hyperaemia, reaction to light), conjunctivae - i.e. conjunctiva of bulbs, lids, and nictitation membrane - (erythema, chemosis), and discharge were recorded as described by DRAIZE, and the aqueous humour (opacity) as described by MCDONALD and SHADDUCK. In addition, any serious lesions or toxic effects other than ocular ones were recorded. The examinations of cornea, iris and aqueous humour were facilitated using optical instruments (e.g. hand slit lamp). To define epithelial damage, one drop of a 1! fluorescein solution was applied to the cornea surface 24 hours after administration of the test substance; where positive effects were recorded this was repeated at the later observation times. The eye was then rinsed with saline to remove excess and non-absorbed fluorescein. Evaluation was performed by means of ultraviolet illumination (area) in a darkened room and diffuse white illumination (intensity), according to MCDONAD and SHADDUCK.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

72 h

Score:

1

Results 2

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

48 h

Score:

0

Results 3

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24 h

Score:

1

Results 4

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Results 5

Irritation parameter:

iris score

Basis:

animal #3

Time point:

72 h

Score:

1

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48 h

Score:

0

Results 7

Irritation parameter:

cornea opacity score

Remarks:

area

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Results 8

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Results 9

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

72 h

Score:

0

Results 10

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

48 h

Score:

0

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24 h

Score:

1

Results 12

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Results 13

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Results 14

Irritation parameter:

cornea opacity score

Remarks:

area

Basis:

animal #2

Time point:

72 h

Score:

0

Results 15

Irritation parameter:

cornea opacity score

Remarks:

area

Basis:

animal #2

Time point:

48 h

Score:

0

Results 16

Irritation parameter:

cornea opacity score

Remarks:

area

Basis:

animal #2

Time point:

24 h

Score:

4

Results 17

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #2

Time point:

72 h

Score:

0

Results 18

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #2

Time point:

48 h

Score:

0

Results 19

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #2

Time point:

24 h

Score:

1

Results 20

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

72 h

Score:

1

Results 21

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

48 h

Score:

0

Results 22

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24 h

Score:

1

Results 23

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

72 h

Score:

2

Results 24

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48 h

Score:

1

Results 25

Irritation parameter:

iris score

Basis:

animal #1

Time point:

72 h

Score:

1

Results 26

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48 h

Score:

0

Results 27

Irritation parameter:

cornea opacity score

Remarks:

area

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Results 28

Irritation parameter:

cornea opacity score

Remarks:

opacity

Basis:

animal #1

Time point:

24/48/72 h

Score:

1

Irritant / corrosive response data:

The application of 0,1 ml of 3,5 -xylidine caused only slight corneal opacity and redding. The symptoms were reversible within 7 days.

Any other information on results incl. tables

animal no. (body weight)	tissue	signs	DRAIZE grade							irritation index
			1h	24h	48h	72h	7d	14d	21d
L27 (3.4 kg)	cornea	o	1	1	1	1	0	-	-	1.0
		a	4	4	4	4	0	-	-
	fluorescein	i	-	1	1	1	0	-	-
		a	-	3c	3c	3c	0	-	-
	iris		0	0	0	1	0	-	-	0.3
	conjunctivae	r	1	1	1	2	0	-	-	1.3
		s	2	1	0	1	0	-	-	0.7
	aqueous humour		0	0	0	0	0	-	-
	discharge		2	2	1	1	0	-	-
L43 (3.6 kg)	cornea	o	1	1	0	0	0	-	-	0.3
		a	4	4	0	0	0	-	-
	fluorescein	i	-	1	1	1	0	-	-
		a	-	2c	2c	2c	0	-	-
	iris		0	0	0	0	0	-	-	0.0
	conjunctivae	r	1	1	1	1	0	-	-	1.0
		s	2	1	0	0	0	-	-	0.3
	aqueous humour		0	0	0	0	0	-	-
	discharge		3	1	0	0	0	-	-
L13 (3.7 kg)	cornea	o	1	1	1	1	0	-	-	1.0
		a	4	4	4	4	0	-	-
	fluorescein	i	-	1	1	1	0	-	-
		a	-	4c	4c	4c	0	-	-
	iris		0	0	0	1	0	-	-	0.3
	conjunctivae	r	1	1	1	1	0	-	-	1.0
		s	2	1	0	1	0	-	-	0.7
	aqueous humour		0	0	0	0	0	-	-
	discharge		2	2	1	2	0	-	-

o=opacity, a=area, i=intensity

r=redness, s=swelling

-=not examined

c=confluent diffuse areas

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Remarks:

Based on EU GHS criteria

Conclusions:

Exposure of the test substance to the eye caused slight reactions of the mucous membranes and the cornea which were more pronounced immediately following administration. Signs proved to be fully reversible within 7days. As there were 2 of 3 tested animals showed a positive response of corneal opacity ≥ 1, test substance can be concluded as Category 2 according to guidanc to regulation (EC) No 1272/2008.

Executive summary:

Eye irritation test was done according to OECD Guideline 405 under GLP compliance. 3 or New Zealand white rabbits' were exposed to the test item (only one eye each, the other one was served as control) for 24 hours. Observation was done 1, 24, 48 and 72 hours and 7days after the exposure. Exposure of the test substance to the eye caused slight reactions of the mucous membranes and the cornea which were more pronounced immediately following administration. Signs proved to be fully reversible within 7days. As there were 2 of 3 tested animals showed a positive response of corneal opacity ≥ 1, test substance can be concluded as Category 2 according to guidanc to regulation (EC) No 1272/2008.