Registration Dossier

Administrative data

Description of key information

Key Studies:
Acute dermal irritation in rabbits: Not classified as skin irritant (MB Research Report number MB 08-17803.03; 2009-15-07)
Acute eye irritation in rabbits; Not classified as an eye irritant (MB Research report number MB 08-17803.04; 2009-21-07)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP, guideline study
Qualifier:
according to
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products Inc., Denver, PA
- Age at study initiation: 4- 5 months
- Weight at study initiation: 2.7 - 3.0 kg
- Housing: individually housed in suspended stainless steel wire bottom cages. Paper bedding was placed beneath the cages and changed at least three times/week.
- Diet (e.g. ad libitum): Fresh PMI Rabbit Chow (Diet #5321)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.05 – 21.94 °C
- Humidity (%): 1.3 – 25.1%
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle

IN-LIFE DATES: From: 1/6/09 To: 1/9/09
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5 ml/site; neat
Duration of treatment / exposure:
4 hours
Observation period:
The test sites were scored for dermal irritation at 60 minutes after removal of wrappings. Reactions were
scored again at 24, 48 and 72 hours following patch removal
Number of animals:
3 (2 males, 1 female)
Details on study design:
TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: 4 ply surgical gauze patch. The patch was secured with non-irritating tape. The torso was wrapped with plastic in a semiocclusive manner and secured with non-irritating tape which completely covered the plastic. This method of wrapping aids in retaining the test patch in position and retards evaporation of volatile substances. The
test article was kept in contact with the skin for 4 hours at which time the wrappings were removed

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test article was removed from the test site by gently washing with distilled water
- Time after start of exposure: At the end of the exposure period, prior to scoring for dermal reactions.

SCORING SYSTEM:

Erythema & Eschar
No erythema 0
Very slight erythema (barely perceptible) 1
Well defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4
Edema
No edema 0
Very slight edema (barely perceptible) 1
Slight edema (edges of area well-defined by definite raising) 2
Moderate edema (raised approximately 1.0 mm) 3
Severe edema (raised more than 1.0 mm, extending beyond the area of exposure) 4
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
other: Study ended at 72 hours
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
other: Study ended at 72 hours
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
other: Study ended at 72 hours
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
other: Study ended at 72 hours
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
other: Study ended at 72 hours
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
other: Study ended at 72 hours
Irritant / corrosive response data:
Erythema was very slight to well defined at 60 minutes and well defined at 24, 48 and 72 hours following the 4 hour exposure. Edema was very slight to moderate at 60 minutes and slight at 24,48 and 72 hours following the 4 hour exposure.

Dermal Observations, Body Weights and Systemic Observations
Rabbit Eartag: H2035 H2017 H2077
Sex: F M M
Pretest Body Weight - kg: 3.0 2.9 2.7
Terminal Body Weight - kg: 2.9 2.9 2.6

Time after patch removal Erythema & Eschar Formation
60 minutes 1t 2t 1t
24 hours 2t 2t 2t
48 hours 2t 2t 2t
72 hours 2t 2t 2
Edema
60 minutes 2 3 1
24 hours 2 2 2
48 hours 2 2 2
72 hours 2 2 2
Systemic Observations
60 minutes A A A
24 hours A A A
48 hours A A A
72 hours A A A
A = Normal t = tape adhesive residue on dose site
Other effects:
There were no abnormal physical signs noted during the observation period.
Two animals lost weight. All other body weight changes were normal.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Not classified as skin irritant
Executive summary:

Since the test article was not expected to produce severe irritation or corrosion, three healthy New Zealand White rabbits (2 males - 1 female) were dosed dermally with trans-ß-farnesene, Lot# KJF-134-53-03, CAS# 18794-84-8. The test article (0.5 ml) was applied dermally to one intact site/rabbit and kept in contact with the skin for 4 hours at which time the wrappings were removed.

Dermal reactions were scored at 60 minutes after removal of wrappings. Reactions were scored again at 24, 48 and 72 hours following patch removal. The skin was also evaluated for ulceration and necrosis or any evidence of tissue destruction at these time periods. Body weights were recorded pretest and at termination.

Erythema was very slight to well defined at 60 minutes and well defined at 24, 48 and 72 hours following the 4 hour exposure. Edema was very slight to moderate at 60 minutes and slight at 24, 48 and 72 hours following the 4 hour exposure. There were no abnormal physical signs noted during the observation period. Two animals lost weight. All other body weight changes were normal.

Dermal application of trans-ß-farnesene caused only mild skin reaction in rabbits following a 4hr semi-occlusive exposure. Skin reactions did not meet the criteria for classification as as a dermal irritant in accordance with EU CLP regulations.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
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: GLP; guideline study
Qualifier:
according to
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source:Covance Research Products, Inc., Denver, PA
- Age at study initiation: 17 weeks
- Weight at study initiation: 2.7 - 2.9 kg
- Housing: individually housed in suspended stainless steel wire bottom cages. Paper bedding was placed beneath the cages and changed at least three times/week
- Diet (e.g. ad libitum): Fresh PMI Rabbit Chow (Diet #5321) was provided daily.
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.77 – 22.77 °C
- Humidity (%): 2.1 - 27%.
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle

IN-LIFE DATES: From:1/6/09 To: 1/9/09
Vehicle:
unchanged (no vehicle)
Controls:
other: contralateral eye served as control
Amount / concentration applied:
0.1 ml in one eye
Duration of treatment / exposure:
Single dose
Observation period (in vivo):
The eyes were examined and scored by the Draize technique at 1, 24, 48 and 72 hours. Sodium fluorescein dye procedures were used at the 24-hour observation interval.
Number of animals or in vitro replicates:
3 (2 males; 1 female)
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): No washing

TOOL USED TO ASSESS SCORE: hand-slit lamp / biomicroscope / fluorescein

A Mini-Maglite® flashlight equipped with a high intensity bulb was used to aid in scoring.
Sodium fluorescein dye procedures were used at the 24-hour observation interval. The eye was
examined with the aid of an ultraviolet light source. Ocular reactions were graded according to the
numerical Draize technique.

SCORING SYSTEM:
SCORING SYSTEM: Ocular reactions were graded according to the numerical Draize technique, summarized below.

SCALE FOR SCORING OCULAR LESIONS (Draize, J. H. et al. J. Pharm. Exp. Ther. 82:377-390, 1944).
(1) CORNEA:
(A) Opacity: Degree of density (area most dense taken for reading):
No ulceration or opacity 0
Scattered or diffuse areas of opacity (other than slight dulling of normal luster),details of iris clearly visible 1(*)
Easily discernible translucent area, details of iris slightly obscured 2(*)
Opalescent areas, no details of iris visible, size of pupil barely discernible 3(*)
Opaque cornea, iris not discernible through the opacity 4(*)
(B) Area of cornea Involved:
One quarter (or less) but not zero 1
Greater than one-quarter, but less than one-half 2
Greater than one-half, but less than three-quarters 3
Greater than three quarters up to whole area 4
SCORE EQUALS A x B x 5 Maximum Total 80

(2) IRIS:
(A) Normal 0
Folds above normal, congestion, swelling, circumcorneal injection (any or all of these or combination
of any thereof), iris still reacting to light (sluggish reaction is positive) 1(*)
No reaction to light, hemorrhage, gross destruction (any or all of these) 2(*)
SCORE EQUALS A x 5 Maximum Total 10

(3) CONJUNCTIVAE:
(A) REDNESS (refers to palpebral and bulbar conjunctivae excluding cornea & iris):
Blood vessels normal 0
Some blood vessels definitely hyperemic (injected) 1
More diffuse, deeper crimson red, individual vessels not easily discernible 2(*)
Diffuse beefy red 3(*)
(B) CHEMOSIS
No swelling 0
Any swelling above normal (includes nictitating 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(*)
(C) DISCHARGE
No Discharge 0
Any amount different from normal (does not include small amounts observed in
inner canthus of normal animals) 1
Discharge with moistening of the lids and hairs just adjacent to lids 2
Discharge with moistening of the lids and hairs and considerable area around the eye 3
SCORE EQUALS (A+B+C)x2 Maximum Total 20

The maximum total score is the sum of all scores obtained for the cornea, iris and conjunctivae.
4.
(*)Indicates a positive response

ULTRAVIOLET FLUORESCEIN SCAN SCORING CODE:

0 = Negative
1 = Positive with an area 1/4 or less
2 = Positive with an area >1/4 but <1/2
3 = Positive with an area >1/2, but <3/4
4 = Positive with an area >3/4, up to entire area

Irritation parameter:
cornea opacity score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable; no opacity
Irritation parameter:
iris score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
0
Reversibility:
other: not applicable
Irritation parameter:
conjunctivae score
Basis:
mean
Time point:
24/48/72 h
Score:
0.33
Max. score:
2
Reversibility:
fully reversible within: 72 hours
Irritation parameter:
chemosis score
Basis:
mean
Time point:
24/48/72 h
Score:
0
Max. score:
1
Reversibility:
fully reversible within: 24 hours
Irritant / corrosive response data:
Ocular Findings, Systemic Observations and Body Weights

An.#/Sex Item Tissue Reading Hour 1 Hour 24 Hour 48 Hour 72
H2116/M A Cornea Opacity 0 0 0 0
B Area 0 0 0 0
1. Total=(AxB)x5 0 0 0 0
C Iris 0 0 0 0
2. Total=Cx5 0 0 0 0
D Conjunctiva Redness 2 1 0 0
E Chemosis 1 0 0 0
F Discharge 0 0 0 0
3. Total=(D+E+F)x2 6 2 0 0

Total=1+2+3 6 2 0 0

Systemic Observations A A A A
Sodium Fluorescein 0 N/A N/A
Pretest Body Weight - 2.9 kg


An.#/Sex Item Tissue Reading Hour 1 Hour 24 Hour 48 Hour 72
H2120/M A Cornea Opacity 0 0 0 0
B Area 0 0 0 0
1. Total=(AxB)x5 0 0 0 0
C Iris 0 0 0 0
2. Total=Cx5 0 0 0 0
D Conjunctiva Redness 2 1 1 0
E Chemosis 1 0 0 0
F Discharge 1 0 0 0
3. Total=(D+E+F)x2 8 2 2 0
Total=1+2+3 8 2 2 0

Systemic Observations A A A A
Sodium Fluorescein 0 N/A N/A
Pretest Body Weight - 2.7 kg

An.#/Sex Item Tissue Reading Hour 1 Hour 24 Hour 48 Hour 72
H2148/F A Cornea Opacity 0 0 0 0
B Area 0 0 0 0
1. Total=(AxB)x5 0 0 0 0
C Iris 0 0 0 0
2. Total=Cx5 0 0 0 0
D Conjunctiva Redness 2 1 0 0
E Chemosis 1 0 0 0
F Discharge 1 0 0 0
3. Total=(D+E+F)x2 8 2 0 0
Total=1+2+3 8 2 0 0
Systemic Observations A A A A
Sodium Fluorescein 0 N/A N/A
Pretest Body Weight - 2.8 kg

A = Normal N/A = not applicable
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Farnesene is not classifiable as an eye irritant per EU CLP criteria.
Executive summary:

Three healthy New Zealand White rabbits (2 males - 1 female), free from evidence of

ocular irritation and corneal abnormalities, were dosed with trans-ß-farnesene, Lot# KJF-134-53-03,

CAS# 18794-84-8. The test article (0.1 ml) was placed into the conjunctival sac of one eye of each

rabbit. The contralateral eye served as a control. The eyes were examined and scored by the Draize

technique at 1, 24, 48 and 72 hours. Sodium fluorescein dye procedures were used at the 24-hour

observation interval. Body weights were recorded pretest.

There was no corneal opacity or iritis noted in any animal at any observation period. Mild conjunctival irritation (redness, swelling and some discharge) noted in the eyes of 3/3 animals 1 hr after treatment had cleared within 72 hours. There were no abnormal physical signs noted during the observation period.

Following instillation into the rabbit eye, trans-beta farnesene provoked only minimal occular irritation. Reactions did not meet the criteria for classification as an eye irritant per EU CLP criteria.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Key guideline studies investigating the skin and eye irritation potential of farnesene are available.

Skin: Erythema (very slight to well defined at 60 minutes and well defined at 24, 48 and 72 hours) was observed in rabbits following a 4 dermal hour exposure. Edema was also was seen (very slight to moderate at 60 minutes and slight at 24,

48 and 72 hours). Skin reactions did not meet the criteria for classification as a dermal irritant

Eye: There was no corneal opacity or iritis noted at in any animal any observation period. Conjunctival irritation (slight redness,swelling and some discharge) was noted in 3/3 eyes (score of 1 at 24 hours), but cleared within 72 hours. Occular reactions did not meet the criteria for classification as an eye irritant.


Justification for selection of skin irritation / corrosion endpoint:
GLP guideline study; Klimisch score of 1

Justification for selection of eye irritation endpoint:
GLP guideline study; Klimisch score of 1

Justification for classification or non-classification

Skin irritation: Reactions did not meet criteria of greater or equal to 2.3 for erythema and for edema in at least 2 or 3 animals.

Eye irritation: Reactions not meet the following criteria; :

– at least in 2 of 3 tested animals, a positive response of:

– corneal opacity ≥ 1 and/or

– iritis ≥ 1, and/or

– conjunctival redness ≥ 2 and/or

– conjunctival oedema (chemosis) ≥ 2

– calculated as the mean scores following grading at 24, 48

and 72 hours after installation of the test material, and which fully reverses within an observation period of 21 days