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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30 June 2014 to 24 July 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not effect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report date:
2014

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 425 (Acute Oral Toxicity: Up-and-Down Procedure)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
up-and-down procedure
Limit test:
yes

Test material

Constituent 1
Reference substance name:
1.25H Farnesene
IUPAC Name:
1.25H Farnesene
Test material form:
other: liquid
Details on test material:
A Certificate of Analysis supplied by the Sponsor is given in Appendix 1 (attachment 1)

Identification: 1.25H Farnesene
Batch: RV-752-153-2
Purity: 98.92%
Physical state / Appearance: clear, colorless liquid
Expiry date: 01 March 2015
Storage Conditions: room temperature in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
Female Wistar (RccHan™:WIST) strain rats were supplied by Harlan Laboratories UK Ltd., Oxon, UK. On receipt the animals were randomly allocated to cages. The females were nulliparous and non-pregnant. After an acclimatization period of at least five days the animals were selected at random and given a number unique within the study which was written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The body weight variation did not exceed ±20% of the body weight of the initially dosed animal.

The animals were individually housed in suspended solid floor polypropylene cages furnished with woodflakes. With the exception of an overnight fast immediately before dosing and for approximately three to four hours after dosing, free access to mains drinking water and food (2014C Teklad Global Rodent diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet, drinking water and bedding were routinely analyzed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 19 to 25 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe.
Doses:
Using available information on the toxicity of the test item, 5000 mg/kg was chosen as the starting dose.
No. of animals per sex per dose:
3 females at 5000 mg/kg
Control animals:
no
Details on study design:
Using available information on the toxicity of the test item, 5000 mg/kg was chosen as the starting dose.

Three individual fasted female animals were treated as follows:

Test Sequence (Animal number) Dose Level, mg/kg Specific Gravity Dose Volume (mL/kg) Short-Term Result
1
(1-0) 5000 0.809 6.19 0
2
(2-0) 5000 0.809 6.19 0
3
(3-0) 5000 0.809 6.19 0

0 = Animal survived

The test was complete after the third animal had been dosed as the following stopping criterion was met:

Three consecutive animals survived at the maximum dose level (5000 mg/kg)

All animals were dosed once only by gavage, using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to the fasted bodyweight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each individual animal to confirm the survival of the previously dosed animals.

The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days.

Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment.

At the end of the observation period the animals were killed by cervical dislocation. All animals were subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.
Statistics:
None

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 95% confidence limits not given in study report.
Mortality:
Individual mortality data are given in Table 1.
There were no deaths.
Clinical signs:
Individual clinical observations are given in Table 1.
No signs of systemic toxicity were noted during the observation period.
Body weight:
Individual body weights and body weight changes are given in Table 2.

All animals showed expected gains in body weight over the observation period.
Gross pathology:
Individual necropsy findings are given in Table 3.
No abnormalities were noted at necropsy.
Other findings:
None

Any other information on results incl. tables

Table 1     Individual Clinical Observations and Mortality Data

Dose Level mg/kg

Animal Number and Sex

Effects Noted After Dosing
(Hours)

Effects Noted During Period After Dosing
(Days)

½

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

5000

1-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0

F2male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-0

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0=   No signs of systemic toxicity

Table 2     Individual Body Weights and Body Weight Changes

Dose Level

mg/kg

Animal Number
and Sex

Body Weight (g) at Day

Body Weight Gain (g) During Week

0

7

14

1

2

5000

1-0 Female

159

170

184

11

14

2-0 Female

160

178

191

18

13

3-0 Female

154

161

173

7

12

Table 3     Individual Necropsy Findings

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

5000

1-0 Female

Killed Day 14

No abnormalities detected

2-0 Female

Killed Day 14

No abnormalities detected

3-0 Female

Killed Day 14

No abnormalities detected

Applicant's summary and conclusion

Interpretation of results:
other: The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be greater than 5000 mg/kg body weight.
Remarks:
Criteria used for interpretation of results: not specified
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be greater than 5000 mg/kg body weight.
Executive summary:

Introduction

The study was performed to assess the acute oral toxicity of the test item in the Wistar strain rat.

 

 

Methods

One fasted female animal was treated with the test item at a dose level of 5000 mg/kg body weight. The animal survived, therefore two additional females were treated sequentially at a dose level of 5000 mg/kg body weight so that a total of three animals were treated. 

 

The test item was administered orally undiluted. Clinical signs and body weight development were monitored during the study. All animals were subjected to gross necropsy.

 

 

Results

Mortality. There were no deaths.

 

Clinical Observations. There were no signs of systemic toxicity.

 

Body Weight. All animals showed expected gains in body weight.

 

Necropsy. No abnormalities were noted at necropsy.

 

 

Conclusion

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was found to be greater than 5000 mg/kg body weight.