Reaction mass of (3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)butan-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)butan-2-one

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed under GLP. All relevant validity criteria were met.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

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)

Remarks:

inspected: July 2012; signature: November 2012

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of (3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)butan-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)butan-2-one

EC Number:

700-956-5

Molecular formula:

not applicable (reaction mass of constitutional isomers)

IUPAC Name:

Reaction mass of (3E)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)but-3-en-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-1-en-1-yl)butan-2-one and 4-(dodecylsulfanyl)-4-(2,6,6-trimethylcyclohex-2-en-1-yl)butan-2-one

Test material form:

gas under pressure: refrigerated liquefied gas

Details on test material:

- Physical state: liquid
- Storage condition of test material: Approximately 4 °C in the dark, under nitrogen.
- Other: Pale yellow

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Recognised animal supplier
- Age at study initiation: 12-20 weeks old
- Weight at study initiation: 2.38 - 2.49 kg
- Housing: individually housed in suspended metal cages
- Diet: certified rabbit food ad libitum
- Water: mains water ad libitum
- Acclimation period: at least 5 days

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

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 mL
- Concentration (if solution): undiluted

Duration of treatment / exposure:

A volume of 0.1 ml of the test material, was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test material, and then released. The left eye remained untreated and was used for control purposes.

Observation period (in vivo):

The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance.

Number of animals or in vitro replicates:

3 males

Details on study design:

The study was performed in a stepwise manner and was started by treatment of a single rabbit. The two other animals were treated in a similar manner after considering the degree of eye irritation observed in the first animal.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): none

SCORING SYSTEM:
The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded:

CORNEAL IRRITATION
Opacity: degree of density (area most dense taken for reading)
No ulceration or opacity (may include slight dulling of normal luster)_____0
Scattered or diffuse areas of opacity, details of iris clearly visible______1
Easily discernible translucent area, details of iris slightly obscured______2
Nacreous area, no details of iris visible, size of pupil barely discernible__3
Opaque cornea, iris not discernible through the opacity______________4

Area of cornea involved:
No ulceration or opacity________________________________________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

IRIS
Normal______________________________________________________0
Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia,
or injection, any of these or combination thereof, iris still reacting to light
(sluggish reaction is positive)____________________________________1
No reaction to light, hemorrhage, gross destruction (any or all of these)___2

CONJUNCTIVAL IRRITATION
Redness (refers to palpebrae and sclera, excluding cornea and iris):
Blood vessels normal__________________________________________0
Some blood vessels definitely hyperaemic (injected)__________________1
Diffuse, crimson color, individual vessels not easily discernible_________2
Diffuse beefy red_____________________________________________3

Chemosis (refers to lids and/or nictitating membranes):
No swelling__________________________________________________0
Any swelling above normal (includes nictitating membranes)___________1
Obvious swelling with partial eversion of the lids____________________2
Swelling with lids about half closed_______________________________3
Swelling with lids more than half closed ___________________________4

Discharge:
No discharge (may include small amounts observed in inner canthus of normal animals)_____0
Any amount different from normal and/or lacrimation_________________________________1
Discharge with moistening of the lids and hairs just adjacent to lids_____________________2
Discharge with moistening of the lids and hairs (considerable area around the eyes)_______3

TOOL USED TO ASSESS SCORE: Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 3

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24/48/72 h

Score:

0.8

Max. score:

3

Reversibility:

fully reversible within: 72h

Results 4

Irritation parameter:

chemosis score

Basis:

mean

Time point:

24/48/72 h

Score:

0.6

Max. score:

4

Reversibility:

fully reversible within: 72h

Results 5

Irritation parameter:

cornea opacity score

Basis:

animal #1

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 6

Irritation parameter:

iris score

Basis:

animal #1

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Remarks:

mean

Time point:

24/48/72 h

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 72h

Results 8

Irritation parameter:

chemosis score

Basis:

animal #1

Remarks:

mean

Time point:

24/48/72 h

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 72h

Results 9

Irritation parameter:

cornea opacity score

Basis:

animal #2

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 10

Irritation parameter:

iris score

Basis:

animal #2

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 11

Irritation parameter:

conjunctivae score

Basis:

animal #2

Remarks:

mean

Time point:

24/48/72 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72h

Results 12

Irritation parameter:

chemosis score

Basis:

animal #2

Remarks:

mean

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 72h

Results 13

Irritation parameter:

cornea opacity score

Basis:

animal #3

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Results 14

Irritation parameter:

iris score

Basis:

animal #3

Remarks:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Results 15

Irritation parameter:

conjunctivae score

Basis:

animal #3

Remarks:

mean

Time point:

24/48/72 h

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72h

Results 16

Irritation parameter:

chemosis score

Basis:

animal #3

Remarks:

mean

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 72h

Irritant / corrosive response data:

No corneal effects were noted. Iridial inflammation was noted in one treated eye one hour after treatment. Moderate conjunctival irritation was noted in all treated eyes one hour after treatment. Moderate conjunctival irritation was noted in one treated eye with minimal conjunctival irritation noted in two treated eyes at the 24-Hour observation. Minimal conjunctival irritation was noted in all treated eyes at the 48-Hour observation. All treated eyes appeared normal at the 72-Hour observation.

Any other information on results incl. tables

Table 1. Individual scores and mean scores for 24, 48 and 72 hours

Organism number and sex	Time after treatment	Corneal Opacity	Iridial Inflammation	Conjunctival Redness	Conjunctival Chemosis
1 (male)	1h	0	0	2	2
	24 h	0	0	2	1
	48 h	0	0	1	0
	72 h	0	0	0	0
Mean (24 – 72 h)		0.00	0.00	1.0	0.3
2 (male)	1h	0	0	2	1
	24 h	0	0	1	1
	48 h	0	0	1	1
	72 h	0	0	0	0
Mean (24 – 72 h)		0.00	0.00	0.7	0.7
3 (male)	1h	0	1	2	2
	24 h	0	0	1	1
	48 h	0	0	1	1
	72 h	0	0	0	0
Mean (24 – 72 h)		0.00	0.00	0.7	0.7

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

EU criteria not met

Conclusions:

Under the conditions of this study the substance is not irritating to the eye.

Executive summary:

The study was performed to OECD TG 405 and EU Method B.5 under GLP to assess the irritancy potential of the test material to the eye following a single application in the New Zealand White rabbit. A volume of 0.1 ml of the test material was placed into the conjunctival sac of one eye of 3 animals. The other eye remained untreated and was used for control purposes. Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment. A single application of the test material to the non-irrigated eye of three rabbits produced no corneal effects, iridial inflammation was noted in one treated eye one hour after treatment only, with moderate conjunctival irritation noted in all treated eyes one hour after treatment with moderate to minimal conjunctival irritation noted in all treated eyes at the 24-hour observation. This was minimal at 48 -hours and which fully reversed at 72-hours. All treated eyes appeared normal at the 72-hour observation. Under the conditions of this study, the test substance is not considered to be irritating to the eye.