Registration Dossier

Administrative data

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

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guidelineopen allclose all
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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
aerosol dispenser: not specified
Remarks:
migrated information: aerosol
Details on test material:
- Name of test material (as cited in study report): trans-beta-farnesene
-CAS number: 18794-84-8

Test animals

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)

Administration / exposure

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.

Results and discussion

Effect levelsopen allclose all
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.

Applicant's summary and conclusion

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.