Reaction product of m-tolylidene diisocyanate and cyclohexylamine and cyclohex-1,2-ylenediamine and (Z)-octadec-9-enylamine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 September 2009 to 25 September 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The study Richeux (2009a) was conducted according to the protocol OECD No. 405 as the study laboratory at the time stated that no alternative in vitro method existed that would be adequate for the purposes of classification labelling. This is stated in the final report for the study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

24th April 2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1-cyclohexyl-3-[2-methyl-5-({[(9Z)-nonadec-9-en-1-yl]carbamoyl}amino)phenyl]urea; 3-{2-[({3-[(cyclohexylcarbamoyl)amino]-2-methylphenyl}carbamoyl)amino]cyclohexyl}-1-[5-({[(7Z)-heptadec-7-en-1-yl]carbamoyl}amino)-2-methylphenyl]urea

EC Number:

945-075-9

Molecular formula:

Mixture of C47H74N8O4 and C34H58N4O2

IUPAC Name:

1-cyclohexyl-3-[2-methyl-5-({[(9Z)-nonadec-9-en-1-yl]carbamoyl}amino)phenyl]urea; 3-{2-[({3-[(cyclohexylcarbamoyl)amino]-2-methylphenyl}carbamoyl)amino]cyclohexyl}-1-[5-({[(7Z)-heptadec-7-en-1-yl]carbamoyl}amino)-2-methylphenyl]urea

Test material form:

solid

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elevage de Gérome, Quartier Labaste, F40260 Linxe
- Age at study initiation: 11 or 18 weeks old
- Weight at study initiation: 2.21 to 3.63 kg
- Housing: Individual box
- Diet (e.g. ad libitum): SDS – C15 provided ad libitum
- Water (e.g. ad libitum): Tap water from public distribution system provided ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): Approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light : 12 hours dark

IN-LIFE DATES: Not reported

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 g per eye of test animal

Duration of treatment / exposure:

72 hours for two animals, 4 days for one animal

Observation period (in vivo):

72 hours for two animals, 4 days for one animal

Number of animals or in vitro replicates:

3

Details on study design:

TEST ANIMALS
- Source: Elevage de Gérome, Quartier Labaste, F40260 Linxe
- Age at study initiation: 11 or 18 weeks old
- Weight at study initiation: 2.21 to 3.63 kg
- Housing: Individual box
- Diet (e.g. ad libitum): SDS – C15 provided ad libitum
- Water (e.g. ad libitum): Tap water from public distribution system provided ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): Approximately 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light : 12 hours dark

IN-LIFE DATES: Not reported

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Results 2

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.7

Results 3

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.3

Results 4

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.3

Results 6

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.7

Results 7

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Results 8

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Results 9

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Results 10

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Results 11

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Results 12

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Results 13

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

other: 1h

Score:

12.3

Reversibility:

fully reversible

Results 14

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

24 h

Score:

4

Reversibility:

fully reversible

Results 15

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

48 h

Score:

2.7

Reversibility:

fully reversible

Results 16

Irritation parameter:

overall irritation score

Basis:

mean

Time point:

72 h

Score:

0.7

Reversibility:

fully reversible

Results 17

Irritation parameter:

overall irritation score

Basis:

animal #2

Remarks:

Only animal #2 assessed at 4d

Time point:

other: 4d

Score:

0

Irritant / corrosive response data:

No clinical signs related to the administration of the test item were observed. The ocular reactions observed during the study were slight to moderate and totally reversible in three animals:
- at the conjunctivae level; a slight to moderate redness noted 1 hour after test item instillation and totally reversible between day 3 and day 4, associated with a slight to moderate chemosis, noted 1 hour after the test item instillation and totally reversible between day 1 and 3.
- at the iris level; a congestion noted 1 hour after the test item instillation in only one animal.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: One eye left untreated as negative control; negative control results were not reported.

Any other information on results incl. tables

Table 1: Individual and mean scores for conjunctivae, iris and cornea

Animal #	Time after Treatment	Conjunctivae		Iris	Cornea
		Chemosis (A)	Redness (C)	Lesion (D)	Opacity (E)
A9776	24 hours	0	1	0	0
	48 hours	0	1	0	0
	72 hours	0	0	0	0
Total		0	2	0	0
Mean		0.0	0.7	0.0	0.0
A9833	24 hours	1	2	0	0
	48 hours	1	1	0	0
	72 hours	0	1	0	0
Total		2	4	0	0
Mean		0.7	1.3	0.0	0.0
A9834	24 hours	1	1	0	0
	48 hours	0	1	0	0
	72 hours	0	0	0	0
Total		1	2	0	0
Mean		0.3	0.7	0.0	0.0
Classification in accordance with the European Regulation		According to the calculated means, the item must not be classified
Classification in accordance with GHS regulation		According to the calculated means, the item must not be classified

Table 2: Determination of ocular irritation indicies

Observation Time	Total Individual Data						Total Score	Average Irritation Index
	Animal #	Weight (kg)	Animal #	Weight (kg)	Animal #	Weight (kg)
	A9776	Start: 3.51	A9833	Start: 2.21	A9834	Start: 2.32
		End: 3.63		End: 2.31		End: 2.44
1 Hour (D0)	6		15		16		37	12.3
24 Hours (D1)	2		6		4		12	4.0
48 Hours (D2)	2		4		2		8	2.7
72 Hours (D3)	0		2		0		2	0.7
Day 4 (D4)			0				0	0.0
Maximum Ocular Irritation Index (Max. O.I.)							12.3
Classification according to the classification established in the Journal Officiel de la Republique Francaise dated July 10th, 1992							Slightly irritant

Table 3: Total and individual scores of ocular irritation for Animal A9776

Observation Time	Conjunctivae				Iris		Cornea			Individual irritation index
	A	B	C	(A+B+C)*2	D	D*5	E	F	E*F*5
				X =		Y =			Z =	X+Y+Z =
1 Hour (D0)	1	1	1	6	0	0	0	0	0	6
24 Hours (D1)	0	0	1	2	0	0	0	0	0	2
48 Hours (D2)	0	0	1	2	0	0	0	0	0	2
72 Hours (D3)	0	0	0	0	0	0	0	0	0	0

Table 4: Total and individual scores of ocular irritation for Animal A9833

Observation Time	Conjunctivae				Iris		Cornea			Individual irritation index
	A	B	C	(A+B+C)*2	D	D*5	E	F	E*F*5
				X =		Y =			Z =	X+Y+Z =
1 Hour (D0)	2	1	2	10	1	5	0	0	0	15
24 Hours (D1)	1	0	2	6	0	0	0	0	0	6
48 Hours (D2)	1	0	1	4	0	0	0	0	0	4
72 Hours (D3)	0	0	1	2	0	0	0	0	0	2
Day 4 (D4)	0	0	0	0	0	0	0	0	0	0

Table 5: Total and individual scores of ocular irritation for Animal A9834

Observation Time	Conjunctivae				Iris		Cornea			Individual irritation index
	A	B	C	(A+B+C)*2	D	D*5	E	F	E*F*5
				X =		Y =			Z =	X+Y+Z =
1 Hour (D0)	3	3	2	16	0	0	0	0	0	16
24 Hours (D1)	1	0	1	4	0	0	0	0	0	4
48 Hours (D2)	0	0	1	2	0	0	0	0	0	2
72 Hours (D3)	0	0	0	0	0	0	0	0	0	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The eye irritation potential of the test item was determined in a guideline study with New Zealand White Rabbits. The ocular reactions observed during the study were slight to moderate and totally reversible in all three animals. The results of the eye irritation study were interpreted according to the European directives 2001/59/E.C and 99/45/E.C. and in accordance with the Globally Harmonised System (Regulation (EC) No 1272/2008) and the test item must not be classified for eye irritation.

Executive summary:

The eye irritation potential of the test item was investigated in an OECD 405 and EC method B.5 guideline study with three albino white rabbits. The test item was grinded and then 0.1 g was applied to one eye of the test animal, with the other eye left untreated as a control. Observations were made after 1 hour, and then after 24, 48 and 72 hours using an opthalmoscope. Eye irritation was evaluated using a scoring system based on chemosis, discharge, redness, iris condition, degree of opacity of the cornea and extent of opacity of the cornea at each observation interval and the total and average irritation index scores were calculated. The results of the eye irritation study were interpreted according to the European directives 2001/59/E.C and 99/45/E.C. and in accordance with the Globally Harmonised System (Regulation (EC) No 1272/2008.

The ocular reactions observed during the study were slight to moderate and totally reversible in all three animals. The results of the eye irritation study were interpreted according to the European directives and in accordance with the Globally Harmonised System and the test item must not be classified for eye irritation.

This study is a GLP compliant guideline experimental study, acceptable without restriction for assessment of this endpoint.