Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was performed between 30 July 2008 and 27 August 2008.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Date of inspection:21/08/2007 Date of signature: 15/10/2007
Test type:
fixed dose procedure
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material:
Oxooil LS9

- Physical state:
Clear colourless liquid

- Storage condition of test material:
Room temperature in the dark

-Date received:
14 July 2008

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
Harlan UK Limited, Bicester, Oxon, UK

- Age at study initiation:
At the start of the study the animals were eight to twelve weeks of age

- Weight at study initiation:
The bodyweight variation did not exceed ± 20% of the initial/mean bodyweight of any previously dosed animal(s)

- Fasting period before study:
Overnight fast immediately before dosing

- Housing:
The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes.
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.

- Diet (e.g. ad libitum):
free access to food (Certified Rat and Mouse Diet) was allowed throughout the study

- Water (e.g. ad libitum):
free access to mains drinking water was allowed throughout the study

- Acclimation period:
At least five days

The diet, drinking water and bedding were routinely analysed and were considered not to contain any contaminants that would reasonably be expected to affect the purpose or integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):
The temperature was set to achieve limits of 19 to 25°C

- Humidity (%):
The relative humidity was set to achieve limits of 30 to 70%

- Air changes (per hr):
The rate of air exchange was at least fifteen changes per hour

- Photoperiod (hrs dark / hrs light):
The lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.


IN-LIFE DATES: From:Day 0 To: Day 14

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle:
For the purpose of the 300 mg/kg dose level the test material was freshly prepared, as required, as a solution in arachis oil BP. For the purpose of the 2000 mg/kg dose level the test material was used as supplied

- Amount of vehicle (if gavage):
The volume administered to each animal was calculated according to the fasted bodyweight at the time of dosing.

- Justification for choice of vehicle:
Arachis oil BP was used because the test material did not dissolve/suspend in distilled water.

- Lot/batch no. (if required):
Not applicable

- Purity:
Determination by analysis of the concentration, homogeneity and stability of the test material preparations was not appropriate because it was not specified in the Study Plan and is not a requirement of the Test Guideline.

MAXIMUM DOSE VOLUME APPLIED:
2000 mg/kg bodyweight


DOSAGE PREPARATION (if unusual):
Not applicable

- Rationale for the selection of the starting dose:
In the absence of data regarding the toxicity of the test material, 300 mg/kg was chosen as the starting dose.
Doses:
Sighting Test: 300 mg/kg and 2000 mg/kg
Main Test: 2000 mg/kg
No. of animals per sex per dose:
Sighting Test:
1 Female rat at 300 mg/kg
1 Female rat at 2000 mg/kg
Main Test:
4 Female rats at 2000 mg/kg
Control animals:
no
Details on study design:
- Duration of observation period following administration:
Clinical observations were made ½, 1, 2, and 4 hours after dosing and then daily for up to fourteen days. Morbidity and mortality checks were made twice daily.

- Frequency of observations and weighing:
Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14 or at death.

- Necropsy of survivors performed:
yes

- Other examinations performed:
behavioural and clinical observations, gross lesions, bodyweight changes, mortality and any other toxicological effects
Statistics:
Not recorded

Results and discussion

Preliminary study:
Individual clinical observations and mortality data are given in Table 1

Mortality
There was no mortality.

Clinical Observations
No signs of systemic toxicity were noted during the observation period.

Bodyweight
Individual bodyweights and bodyweight changes are given in Table 2.
The animal showed expected gains in bodyweight over the observation period.

Necropsy
Necropsy findings are given in Table 3.
No abnormalities were noted at necropsy.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Remarks on result:
other: CL not recorded
Mortality:
One animal was found dead twelve days after dosing.
Clinical signs:
Signs of systemic toxicity noted in one animal during the observation period were hunched posture, pilo-erection, tiptoe gait, reduced respiratory rate and noisy respiration.
Body weight:
Individual bodyweights and bodyweight changes are given in Table 5.
The surviving animals showed expected gains in bodyweight over the observation period.

Gross pathology:
Individual necropsy findings are given in Table 6.
Abnormally red lungs, dark liver and dark kidneys were were noted at the necropsy of the animal that was found dead twelve days after dosing.

Other findings:
NONE

Any other information on results incl. tables

Table1               Individual Clinical Observations and Mortality Data -300mg/kg

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

300

1-0

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

Table2               Individual Bodyweights and Bodyweight Changes -300mg/kg

Dose Level

mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight Gain (g)
During Week

0

7

14

1

2

300

1-0 Female

166

186

192

20

6

Table3               Necropsy Findings -300 mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

300

1-0 Female

Killed Day 14

No abnormalities detected

Table4               Individual Clinical Observations and Mortality Data -2000mg/kg

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

2000

2-0

Female

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

3-1

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-2

Female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

3-3

Female

H Rd

H Rd

H Rd

H P

H P

0

0

0

0

0

0

Rn

Rn

H P Rn

H P Rn Wt

X

 

 

 


0= No signs of systemic toxicity                     Rn = Noisy respiration

H = Hunched posture                                         Wt = Tiptoe gait

Rd = Reduced respiratory rate                          X = Found dead

P = Pilo-erection

Table5               Individual Bodyweights and Bodyweight Changes-2000mg/kg

Dose Level

mg/kg

Animal Number and Sex

Bodyweight (g) at Day

Bodyweight (g)
at Death

Bodyweight Gain (g) During Week

0

7

14

1

2

2000

2-0 Female

158

177

196

 

19

19

3-0 Female

171

178

196

 

7

18

3-1 Female

176

190

217

 

14

27

3-2 Female

170

174

194

 

4

20

3-3 Female

182

186

-

144

4

-

Table6               Individual Necropsy Findings-2000mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

2-0 Female

Killed Day 14

No abnormalities detected

3-0 Female

Killed Day 14

No abnormalities detected

3-1 Female

Killed Day 14

No abnormalities detected

3-2 Female

Killed Day 14

No abnormalities detected

3-3 Female

Found dead Day 12

Abnormally red lungs

Dark liver

Dark kidneys

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Sprague Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight.
Executive summary:

Introduction.  The study was performed to assess the acute oral toxicity of the test material in the Sprague‑Dawley CD strain rat. The method was designed to meet the requirements of the following:

§        OECD Guidelines for Testing of Chemicals No 420 “Acute Oral Toxicity - Fixed Dose Method” (2001)

§        Method B1bisAcute Toxicity (Oral) of Commission Directive 2004/73/EC

Method. 

Following a sighting test at dose levels of 300 mg/kg and2000 mg/kg, a further group of four fasted females was given a single oral dose of undiluted test material at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight were monitored during the study. All animals were subjected to gross necropsy.

Mortality. 

One animal treated at a dose level of 2000 mg/kg was found dead twelve days after dosing. 

Clinical Observations. 

Signs of systemic toxicity noted in one animal treated at a dose level of 2000 mg/kg were hunched posture, pilo-erection, tiptoe gait, reduced respiratory rate and noisy respiration. 

Bodyweight. 

Surviving animals showed expected gains in bodyweight.

Necropsy. 

Abnormally red lungs, dark liver and dark kidneys were seen in the animal treated at a dose level of 2000mg/kg that was found dead twelve days after dosing.

Conclusion. The acute oral median lethal dose (LD50) of the test material in the female Sprague‑Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight . The clinical signs and necropsy observations of the animal that died on Day 12 of the observation period were consistent with partial aspiration of the dose material and not to toxicological effects.