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Diss Factsheets

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was conducted between 17 January 2012 and 09 February 2012.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2012
Report date:
2012

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Reference substance name:
Reaction mass of 2,4-dimethyl-6-phenyl-3,6-dihydro-2H-pyran and 2-methyl-4-methylene-6-phenyltetrahydro-2H-pyran and 4,6-dimethyl-2-phenyl-3,6-dihydro-2H-pyran
Molecular formula:
C13H160
IUPAC Name:
Reaction mass of 2,4-dimethyl-6-phenyl-3,6-dihydro-2H-pyran and 2-methyl-4-methylene-6-phenyltetrahydro-2H-pyran and 4,6-dimethyl-2-phenyl-3,6-dihydro-2H-pyran
Test material form:
other: Liquid
Details on test material:
Sponsor's identification: Pelargene
Description: Clear colourless liquid
Batch number: NX00002642
Purity: 99.03%
Date received: 08 November 2011
Expiry date: 14 August 2012
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 animals were nulliparous and non-pregnant. After an acclimatisation period of at least five days the animals were selected at random and given a number unique within the study by indelible ink-marking on the tail and a number written on a cage card. At the start of the study the animals were eight to twelve weeks of age. The bodyweights fell within an interval of ±20% of the mean initial bodyweight of the first treated group.
The animals were housed in groups of three in suspended solid-floor polypropylene cages furnished with wood flakes. 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 analysed 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. Any occasional deviations from these targets were considered not to have affected the purpose or integrity of the study. 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)
Doses:
2000 mg/kg
No. of animals per sex per dose:
Six females per dose. Treatment of animals was sequential. Sufficient time was allowed between each group to confirm the survival of the previously dosed animals.
Control animals:
no
Details on study design:
Using available information on the toxicity of the test item, 2000 mg/kg was chosen as the starting dose.

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 group to confirm the survival of the previously dosed animals.
The animals were observed for deaths or overt signs of toxicity 0.5 1, 2 and 4 hours after dosing and subsequently once daily for up to fourteen days.
Individual bodyweights were recorded prior to dosing and seven and fourteen days after treatment or at death.
At the end of the observation period the surviving 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.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
One animal was killed for humane reasons, one day after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence.
Clinical signs:
Signs of systemic toxicity noted were hunched posture, increased salivation, decreased respiratory rate, laboured or noisy respiration, ataxia, pilo-erection and lethargy. Hypothermia and chromodacryorrhea were confined to one animal. Surviving animals appeared normal two or four days after dosing.
Body weight:
Surviving animals showed expected gains in bodyweight over the study period.
Gross pathology:
Abnormalities noted at necropsy of the animal that was killed during the study were dark liver, dark kidneys and epithelial sloughing of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Any other information on results incl. tables

Individual clinical observations and mortality data

Dose level

(mg/kg)

Animal No. & sex

Effects noted after dosing

(hours)

Effects noted during period after dosing

(days)

0.5

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2000

1-0 female

HS

0

0

0

A

0

0

0

0

0

0

0

0

0

0

0

0

0

1-1 female

HSRn

S

S

0

A

0

0

0

0

0

0

0

0

0

0

0

0

0

1-2 female

HS

0

0

0

A

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0 female

0

0

0

0

HAPHol ChRIRdX*

 

 

 

 

 

 

 

 

 

 

 

 

 

2-1 female

0

0

0

0

HA

HARd

H

0

0

0

0

0

0

0

0

0

0

0

2-2female

0

0

0

0

HALP

HARd

H

0

0

0

0

0

0

0

0

0

0

0

0 = No signs of systemic toxicity

H = Hunched posture

S = Increased salivation

Rd = Decreased respiratory rate

Rl = Laboured respiration

Rn = Noisy respiration

A = Ataxia

P = Piloerection

L = Lethargy

Ho = Hypothermia

Ch = Chromodacryorrhea

X* = Animal killed for humane reasons due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project licence.

 

Individual body weights and weekly body weight changes

Dose level

(mg/kg)

Animal No. & sex

Bodyweight (g) at day

Bodyweight (g) at death

Bodyweight gain (g) during week

0

7

14

1

2

2000

1-0 female

158

184

195

 

26

11

1-1 female

159

195

207

 

36

12

1-2 female

156

184

202

 

28

18

2-0 female

163

-

-

153

-

-

2-1 female

157

180

195

 

23

15

2-2female

160

180

200

 

20

20

- = Animal dead

 

Individual necropsy findings

Dose level

(mg/kg)

Animal No. & sex

Time of Death

Macroscopic Observations

2000

1-0 female

Killed Day 14

No abnormalities detected

1-1 female

Killed Day 14

No abnormalities detected

1-2 female

Killed Day 14

No abnormalities detected

2-0 female

Humanely killed Day 1

Liver: Dark

Kidneys: Dark

Gastric mucosa: Epithelial sloughing

2-1 female

Killed Day 14

No abnormalities detected

2-2female

Killed Day 14

No abnormalities detected

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar 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 item following a single oral administration in the Wistar strain rat. The method was designed to be compatible with the following:

§OECD Guidelines for the Testing of Chemicals No. 423 “Acute Oral Toxicity – Acute Toxic Class Method” (adopted 17 December 2001)

§Method B1 Acute Toxicity (Oral) of Commission Regulation (EC) No. 440/2008

 

Method.

A group of three fasted females was treated with the test item at a dose level of 2000 mg/kg bodyweight. This was followed by a further group of three fasted females at the same dose level.

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

Mortality.

One animal was killed for humane reasons, one day after dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence.

Clinical Observations.

Signs of systemic toxicity noted were hunched posture, increased salivation, decreased respiratory rate, laboured or noisy respiration, ataxia, pilo-erection and lethargy. Hypothermia and chromodacryorrhea were confined to one animal. Surviving animals appeared normal two or four days after dosing.

Bodyweight.

Surviving animals showed expected gains in bodyweight over the study period.

Necropsy.

Abnormalities noted at necropsy of the animal that was killed during the study were dark liver, dark kidneys and epithelial sloughing of the gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Conclusion.

The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bodyweight.

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