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Administrative data

Description of key information

Key studies were completed for each of the three exposure routes ( oral, inhalation and dermal) for the substance under GLP conditions and in accordance or equivalent to EU and OECD methods.  
There were no mortalities in the studies. The acute oral and dermal LD50 values were > 5000 mg/kg and the inhalation LC50 (aerosol) was > 2060 mg/m3

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
This study was designed to comply with the standards set forth in EPA Health Effects Testing Guidelines, OPPTS Series 870.1100,December 2002 and in OECDGuidelines for the testing of Chemicals, Guideline 425 updated March 2006. The study was also conducted in accordance with Good Laboratory Practices requirements of EPA, 40 CFR 160 and 792, FDA 21 CFR 58, and the OECD, Principles on Good Laboratory Practices, 1997.
Qualifier:
according to
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
GLP compliance:
yes
Test type:
up-and-down procedure
Limit test:
yes
Species:
rat
Strain:
other: Wistar Albino
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Ace Animals; Boyertown, Pennsylvanina USA
- Age at study initiation: 10 weeks
- Weight at study initiation: 209-218 grams
- Fasting period before study: 16-20 hours prior to dosing
- Housing: individually housed in suspended stainless steel wire bottom cages; paper bedding beneath cages with bedding changed 3x/week.
- Diet (e.g. ad libitum): PMI rat chow ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 24.38 °C
- Humidity (%): 0 - 3.6%
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

IN-LIFE DATES: From: 12/30/2008 To: 1/19/2009
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5000 mg/kg

- Rationale for the selection of the starting dose: Initially one animal was dosed. Since it survived, 2 additional animals were dosed.
Doses:
5000 mg/kg
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: weighing: pre-test, weekly, and at termination. Observation: Daily for toxicological effects and twice daily for mortality
- Necropsy of survivors performed: yes
- Other examinations performed: body weight and systemic toxicity
Preliminary study:
First rat was dosed at 5000 mg/kg. No mortality so proceeded with dosing other rats with 5000 mg/kg
Sex:
female
Dose descriptor:
approximate LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
95% CL:
> 5 000
Mortality:
None observed
Clinical signs:
piloerection, wettness and staining about the urogenital area, alopecia, dark substance on nose.
Body weight:
Dose Volume and Body Weights
Dose: 5000 mg/kg
Dose Volume
An. # & Sex in cc Day 0 Day 7 Day 14
1 F 1.30 218 260 279
2 F 1.30 213 275 273
3 F 1.30 209 258 267

MEAN 213 264 273
S.D. 4.5 9.3 6.0
# 3 3 3
Gross pathology:
No findings in necropsy

Systemic Observations
DOSE 5000 mg/kg

Animal # / Sex 1/F 2/F 3/F
15 minutes
Hour 1
Hour 2
Hour 4
Day 1 R T,F,1 T,F,1
Day 2 R T,1
Day 3 2
Day 4 2
Day 5 2
Day 6 2
Day 7 2
Day 8 2
Day 9 2
Day 10 2
Day 11 2 2
Day 12 2 2,3
Day 13 2 2,3
Day 14 2 2
No entry indicates animal appeared normal at that observation period.
F = Piloerection R = wetness of the anogenital area T = soiling of the anogenital area 1 = anogenital area stained yellowish around back tail area
2 = alopecia around anogenital area 3 = dark substance on nose
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Rat Oral LD50 > 5000 mg/kg
Executive summary:

Initially, one healthy female Wistar albino rat was dosed orally with trans-ßfarnesene, (Lot# KJF-134-53-03, CAS# 18794-84-8) at 5000 mg/kg. Since the animal survived, two additional animals were dosed at 5000 mg/kg. The rats were observed at 15 minutes, 1, 2 and 4 hours postdose and once daily for 14 days for toxicity and pharmacological effects. All animals were observed twice daily for mortality. Body weights were recorded immediately pretest, weekly and at termination. All animals were examined for gross pathology. The potential for toxicity was based on the mortality response noted.

All three females survived the 5000 mg/kg oral dose. Instances of soiling, wetness and yellow staining of the anogenital area, piloerection, localized alopecia and chromorhinorrhea were noted during the observation period.

Body weight changes were normal in 2/3 animals. One animal lost weight during the second week of the observation period.

There were no macroscopic observations during terminal necropsy.

The LD50 of trans-ß-farnesene following oral administration to the rat was greater than 5000 mg/kg.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
5 000 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
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 870.1300 (Acute inhalation toxicity)
Deviations:
yes
Remarks:
The afternoon mortality observations were not performed for one day of the study. The animals were older than 12 weeks as specified in protocol. Deviations did not have impact on final study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 403 (Acute Inhalation Toxicity)
GLP compliance:
yes
Test type:
fixed concentration procedure
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Ace Animals; Boyertown, PA
- Age at study initiation: 8- 14 weeks old
- Weight at study initiation: 251-378 grams for males; 224-317 grams for females

- Housing: Animals were identified by cage notation and indelible tail marks. The animals were housed 1/cage in suspended cages. Paper bedding was placed beneath the cages and changed at least three times/week
- Diet (e.g. ad libitum): Fresh PMI Rat Chow (Diet #5012) ad libitum except during 4 hour exposure period
- Water (e.g. ad libitum): Ad libitum except during 4 hour exposure period
- Acclimation period:

IN-LIFE DATES: From:9/13/2010 To: 9/27/10 (last day data collected)
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
other: Whole body with nose oriented toward inlet (see illustration section). Following exposure, the animals were gently washed with warm tap water to remove any residual test substance from the face and body.
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Glass chamber
- Exposure chamber volume: 100 Liter
- Method of holding animals in test chamber:
A 100 liter dynamic glass chamber designed to insure uniform spatial distribution of aerosols and which permitted continuous observation during exposure was used. The chamber was partitioned internally with wire screening into a total of ten non-restraining cubicles. One animal was placed in each cubicle.

- Source and rate of air: The airflow through the chamber was calculated to yield at least 10 to 15 air changes per hour so that adequate oxygen was supplied to the animals. The chamber was maintained at a negative pressure differential to the immediate environment in order to keep the test atmosphere contained.

- Method of conditioning air: Chamber temperature and humidity of air entering the chamber were recorded.

- System of generating particulates/aerosols: Farnesene was added from a Harvard Infusion Pump into an atomizing nozzle (Spraying Systems Model 1/8 JBC). The appropriate flow rate was determined pretest. The spray nozzle was powered by pre-filtered compressed air. Nozzle pressure was monitored using a pressure gauge. The exhaust air was passed through filters before entering into a rotameter and vacuum pump.


- Method of particle size determination:

Mass median aerodynamic diameter (MMAD) was calculated pretest and during each exposure period. An 8 stage Andersen cascade impactor was used to determine particle size. Air was drawn through the impactor for ten minutes. The impactor filter paper collection stages were weighed before and after the air sampling to determine the mass collected at each filter paper collection stage. The MMAD was determined graphically using three cycle logarithmic probit paper. The geometric standard deviation was calculated. A pretest MMAD of 4 microns or less was required to ensure that the particles generated during exposure were in the respirable range. Particle size measurements were recorded at least three times during the exposure period. The average particle size was calculated.


- Temperature, humidity, pressure in air chamber:

Time Airflow Temp Negative Pressure Relative Humidity of air entering chamber
(minutes) L/min Degrees C inches H2O %

30 32 (20L/min) 22 0.3 49
60 32 (20L/min) 22 0.3 49
90 32 (20L/min) 22 0.3 49
120 32 (20L/min) 23 0.3 46
150 32 (20L/min) 23 0.3 46
180 32 (20L/min) 23 0.3 44
210 32 (20L/min) 23 0.3 44
240 32 (20L/min) 24 0.3 41



TEST ATMOSPHERE
- Brief description of analytical method used:

The target concentration was determined prior to exposure by determining the best flow rate for generating the desired concentration.

In order to calculate the concentration gravimetrically, the total solid was determined prior to exposure by drying a preweighed sample of the test article for two minutes, reweighing and calculating the total solid: Final weight\Initial weight.

During the exposure, chamber air was drawn through preweighed filters. The filters were removed and reweighed. The actual concentration of the test article was calculated based on the total solid, the amount of test article dispersed and the air flow through the chamber


- Particle size distribution: See attachment to "Background material." Particle size analyses revealed an average mass median aerodynamic diameter of 0.86 micrometers with an average Geometric standard deviation of 3.16 micrometers




Analytical verification of test atmosphere concentrations:
no
Duration of exposure:
ca. 4 h
Concentrations:
2.06 mg/L
No. of animals per sex per dose:
5 males and 5 females
Control animals:
no
Details on study design:
All rats were monitored during the exposure period, one hour after exposure and once daily thereafter for 14 days for toxicity and pharmacological effects. The rats were observed twice daily for mortality. Body weights were recorded prior to exposure, weekly and at termination. All animals were examined for gross pathology.
Sex:
female
Dose descriptor:
LC50
Effect level:
> 2.06 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Sex:
male
Dose descriptor:
LC50
Effect level:
> 2.06 mg/L air
Based on:
test mat.
Exp. duration:
4 h
Mortality:
None observed
Clinical signs:
Piloerection, wetness of the anogenital area, closed eyes, ocular lacrimation/crust formation, and coating of the fur were noted during the exposure. Chromorhinorrhea, piloerection, wetness and soiling of the anogenital area, few faeces, coating of the fur with test article, and localized alopecia were noted during the remainder of the study. Weight loss was noted in 4/10 animals.

TIME PERIODS ANIMAL #/SEX
Day 0=Exposure 1/M 2/M 3/M 4/M 5/M 6/F 7/F 8/F 9/F 10/F
Day 0 -32 Min 1 1 1 1 1
Day 0- 92 Mins 1 1 1 1R 1F 1 1 1 1F 1R
Day 0- 152 Mins 1,6 1 1 1R 1F 1,6 1 1,6 1,F 1,R
Day 0- 212 Mins 1,6 1,4 1 1,R 1,4,F 1,4,6 1 1,4,6 1,F 1,R
Day 0: Hr 5 4 4 4 4 4 4 4 4 4 4
Day 1 4 4 4 4X 4,S 4,S 4 4 4,SRF 4
Day 2 4 4FRT
Day 3 4
Day 4
Day 5
Day 6
Day 7 10
Day 8 10
Day 9 10
Day 10 10
Day 11 10
Day 12 10
Day 13 10,11
Day 14 10,11

No entry indicates animal appeared normal at that observation period. 1 = eyes closed 4 = fur coated with test article
6= eyes crusting/lacrimation, 10 = alopecia localized on abdomen, 11 = alopecia localized on back
F = piloerection R = wetness of the anogenital area , S=chromorhinorrhea, T= soiling of anogenital area
X= few feces
Body weight:
All weights are in grams

Animal # Sex Day 0 Day 7 Day 14
1 M 340 358 384
2 M 346 366 388
3 M 354 369 400
4 M 378 396 430
5 M 251 301 369
Mean 334 358 394
Std Dev 48.5 34.9 22.9
N = 5 5 5
6 F 317 335 330
7 F 257 349 260
8 F 235 254 249
9 F 230 235 239
10 F 224 220 237
Mean 253 279 263
Std Dev 38.1 59.3 38.6
N = 5 5 5
Gross pathology:
none
Other findings:
At necropsy, localized alopecia was noted in one animal.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
There were no deaths following exposure of rats to Farnesene aerosol at a concentration of 2.06 mg/L. The rat 4 hour inhalation LC50 is > 2.06 mg/L
Executive summary:

Five healthy male and five healthy female Sprague Dawley rats were exposed to an aerosol atmosphere of farnesene at a concentration of 2.06 mg/L for 4 hours. Chamber temperature, relative humidity of air entering the chamber, chamber air flow and negative pressure were monitored and recorded. All rats were monitored during the exposure period, one hour after exposure, and once daily thereafter for 14 days for toxicity and pharmacological effects. The rats were observed twice daily for mortality. Body weights were recorded prior to exposure, weekly, and at termination. All animals were examined for gross pathology. Abnormal tissues were preserved in 10% neutral buffered formalin for possible future histologic examination. 

The target concentration was estimated prior to exposure. Actual concentrations were determined gravimetrically during exposure. Particle size analyses revealed an average mass median aerodynamic diameter of 0.86 micrometer with an average geometric standard deviation of 3.16 micrometers. 

 

All animals survived the four hour 2.06 mg/L exposure. Instances of piloerection, wetness of the anogenital area, closed eyes, ocular lacrimation/crust formation and coating of the fur with test article were noted during the exposure. Chromorhinorrhea, piloerection, wetness of the anogenital area, soiling of the anogtenital area, few faeces, coating of the fur with test article and localized alopecia were noted during the remainder of the study.   Body weight changes were normal in 6/10 animals. Instances of weight loss were noted in four females. Necropsy results were normal in 9/10 animals. Localized alopecia was noted in one female. The rat 4 hour inhalation LC50 is > 2.06 mg/L.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
2 060 mg/m³

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
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 870.1200 (Acute Dermal Toxicity)
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products, Inc., Denver, PA
- Age at study initiation: 17 weeks
- Weight at study initiation: The pretest body weight range was 2.4 - 2.7 kg for males and 2.7 - 2.9 kg for females.
- Fasting period before study: No
- Housing: housed 1/cage in suspended wire 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: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 14.72 – 22.94 °C
- Humidity (%): 0-27%
- Air changes (per hr): Not specified
- Photoperiod (hrs dark / hrs light): 12 hour light/dark cycle

IN-LIFE DATES: From: 1/6/09 To: 1/20/09
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 10 cm x 15 cm
- % coverage: 10%
- Type of wrap if used: 4 ply gauze dressing. The torso was wrapped with plastic sheeting in a semi-occlusive manner, which was secured with non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Residual test article was removed by gently washing with distilled water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 5000 mg/kg dose
- Concentration (if solution): neat
- Constant volume or concentration used: yes/no
- For solids, paste formed: Not aplicable

VEHICLE - none
Duration of exposure:
24 hours
Doses:
5000 mg/kg
No. of animals per sex per dose:
5 males and 5 females
Control animals:
not required
Details on study design:
Dermal responses were recorded at 24 hours postdose and on days 7 and 14. Animals were observed for toxicity and pharmacological effects at 1, 2 and 4 hours postdose and once daily for 14 days. All animals were observed twice a day for mortality. Body weights were recorded pretest, weekly and at termination. All animals were examined for gross pathology. Abnormal tissues were preserved in 10% neutral buffered formalin for possible future histological examination.
Preliminary study:
Not applicable
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Sex:
male
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
None observed
Clinical signs:
No systemic observations noted during study.
Body weight:
An. # Sex Dose (cc) Day 0 Day 7 Day 14
H2126 M 16.3 2.7 2.6 2.9
H2118 M 15.1 2.5 2.4 2.6
H2121 M 14.4 2.4 2.3 2.4
H2122 M 15.7 2.6 2.5 2.7
H2125 M 16.3 2.7 2.6 2.8
MEAN 2.6 2.5 2.7
S.D. 0.1 0.1 0.2
# 5 5 5

H2149 F 17.5 2.9 2.7 2.9
H2151 F 16.9 2.8 2.8 2.9
H2153 F 17.5 2.9 2.7 2.9
H2154 F 17.5 2.9 2.8 3.0
H2155 F 16.3 2.7 2.8 2.9
MEAN 2.8 2.8 2.9
S.D. 0.1 0.1 0.0
# 5 5 5
Gross pathology:
An. # Sex 24 hours Day 7 Day 14
Erythema Edema Erythema Edema Erythema Edema
H2126 M 1 0 1p 0 1e 0
H2118 M 1 0 2 2 1ef 1
H2121 M 0 0 1 2 1ef 0
H2122 M 0 0 1 2 1ef 0
H2125 M 1 0 1 2 1ef 1
H2149 F 1 0 1 1 1ef2 0
H2151 F 0 0 1p 1 1ef2 1
H2153 F 1 0 2p 2 1ef2 1
H2154 F 1 0 1 1 1ef2 0
H2155 F 0 0 1 1 1ef 0

f = flaking skin e = alopecia surrounding dose site 2 = alopecia extended down both sides and completely across the abdomen p = pale areas
Other findings:
Upon necropsy, skin abnormalities at treatment sites were noted in all animals.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Rabbit 24 hour dermal LD50 > 5000 mg/kg
Executive summary:

The acute dermal toxicity of farnesene was investigated in rabbits. The test material was applied to the shaved dorsal skin of 5 male and 5 female rabbits at 5000 mg/kg of body weight, under an occlsive dressing. The test article was kept in contact with the skin for 24 hours. Dermal responses were recorded at 24 hours postdose and on days 7 and 14. Animals were observed for toxicity and pharmacological effects at 1, 2 and 4 hours postdose and once daily for 14 days. All animals were observed twice a day for mortality.

Body weights were recorded pretest, weekly and at termination. All animals were examined for gross pathology. Abnormal tissues were preserved in 10% neutral buffered formalin for possible future histological examination.

There were no deaths during the study and all animals survived the 5000 mg/kg dermal application in good health. Dermal responses were absent to very slight at 24 hours, very slight to well defined on day 7 and very slight on day 14. Additionally, pale areas, flaking skin and areas of poor hair regrowth were noted during the study. Slight body weight loss was noted on day 7 in 8/10 animals, but all returned to normal by day 14. Treated skin abnormalities were the only abnormality noted at necropsy.

The dermal LD50 of trans-ß-farnesene, in the rabbit is greater than 5000 mg/kg of body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
5 000 mg/kg bw

Additional information

Well conducted key studies, undertaken to OECD guidelines, are available for the acute oral, dermal and inhalation toxicity of farnesene.

Oral: There were no deaths during the study and the oral LD50 in the rat for trans-ß-farnesene, is greater than 5000 mg/kg of body weight.

Inhalation: There were no deaths following inhalation exposure of rats to farnesene aerosol at a concentration of 2060 mg/m3. Instances of piloerection, wetness of the anogenital area, closed eyes, ocular lacrimation/crust formation and coating of the fur with test article were noted during the exposure. Chromorhinorrhea, piloerection, wetness of the anogenital area, soiling of the anogtenital area, few faeces, coating of the fur with test article and localized alopecia were noted during the remainder of the study. Body weight changes were normal in 6/10 animals. Instances of weight loss were noted in four females. Necropsy results were normal in 9/10 animals. Localized alopecia was noted in one female. The inhalation LC50 was > 2060 mg/m3.

Dermal: No deaths occurred following topical dermal exposure of rabbits to farnesene at a level of 5000 mg/kg. Dermal responses were absent to very slight at 24 hours, very slight to well defined on day 7 and very slight on day 14. Additionally, pale areas, flaking skin and areas of poor hair regrowth were noted during the study. Slight body weight loss was noted on day 7 in 8/10 animals, but all returned to normal by day 14. Treated skin abnormalities were the only abnormality noted at necropsy. The dermal LD50 of trans-ß-farnesene, is greater than 5000 mg/kg of body weight.

In conclusion, based on data available, farnesene is not classified for acute toxicity according to EU CLP regulations


Justification for selection of acute toxicity – oral endpoint
GLP guideline study; Klimisch score of 1

Justification for selection of acute toxicity – inhalation endpoint
GLP guideline study; Klimisch score of 1

Justification for classification or non-classification

Data available do not meet the CLP hazard classification criteria for acute toxicity