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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was conducted between 31 October 2013 and 26 November 2013.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study is considered to be a reliability 1 as it has been conducted according to OECD Test Guideline 423 using the Acute Toxic Class Method and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report date:
2014

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (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
Chemical structure
Reference substance name:
Reaction mass of (5R)-octahydro-1H-4,7-methanoinden-5-ylacetaldehyde and (5S)-octahydro-1H-4,7-methanoinden-5-ylacetaldehyde
EC Number:
940-300-7
Cas Number:
1339119-15-1
Molecular formula:
C12H18O
IUPAC Name:
Reaction mass of (5R)-octahydro-1H-4,7-methanoinden-5-ylacetaldehyde and (5S)-octahydro-1H-4,7-methanoinden-5-ylacetaldehyde
Test material form:
other: liquid
Details on test material:
Identification: IFF TM 11-213
Storage Conditions: Approximately 4 °C in the dark

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
Animals and Animal Husbandry

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 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 body weight variation did not exceed ± 20 % of the body weight of the initially dosed animal.

The animals were housed in groups of up to three 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. 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:
arachis oil
Details on oral exposure:
Test Item Formulation and Experimental Preparation
For the purpose of the 2000 mg/kg dose level, the test item was used as supplied. The specific gravity was determined and used to calculate the appropriate dose volume for the required dose level. For the purpose of the 300 mg/kg dose level, the test item was freshly prepared, as required, as a solution in arachis oil BP. Arachis oil BP was used because the test item did not dissolve/suspend in distilled water.

The test item was formulated within two hours of being applied to the test system. It is assumed that the formulation was stable for this duration.

No analysis was conducted to determine the homogeneity, concentration or stability of the test item formulation. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.


Doses:
A group of three fasted females was treated with the test item at a dose level of 300 mg/kg body weight. Based on the results from this dose level, further groups of fasted females were treated at dose levels of 300 or 2000 mg/kg body weight. Dosing was performed sequentially.

The test item was administered orally undiluted at a dose level of 2000 mg/kg and as a solution in arachis oil BP at a dose level of 300 mg/kg.

No. of animals per sex per dose:
3 female rats per dose level
Control animals:
no
Details on study design:
Procedure
In the absence of data regarding the toxicity of the test item, 300 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 body weight 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 body weights 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.


Data evaluations included the relationship, if any, between the exposure of the animal to the test item and the incidence and severity of all abnormalities including behavioral and clinical observations, gross lesions, body weight changes, mortality and any other toxicological effects.

Using the mortality data obtained, an estimate of the acute oral median lethal dose (LD50) of the test item was made.

The results were also evaluated according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
500 mg/kg bw
Based on:
test mat.
Mortality:
Animals treated at a dose level of 2000 mg/kg were killed for humane reasons, during the day of dosing, due to the occurrence of clinical signs of toxicity that exceeded the severity limit set forth in the UK Home Office Project Licence or found dead one day after dosing. One of the second group of animals, treated at a dose level of 300 mg/kg, was found dead one day after dosing.
Clinical signs:
Signs of systemic toxicity noted at a dose level of 2000 mg/kg were ataxia, lethargy, hunched posture, prostration, labored respiration, decreased respiratory rate, occasional body tremors, hypothermia and pallor of the extremities. Hunched posture was noted in one animal treated at a dose level of 300 mg/kg.
Body weight:
Surviving animals showed expected gains in body weight over the observation period.

Gross pathology:
Abnormalities noted at necropsy of animals that were humanely killed or died during the study were pale liver, patchy pallor of the liver, pale or dark kidneys, clear liquid present in the stomach, hemorrhage and epithelial sloughing of the gastric mucosa and hemorrhage of the non-glandular epithelium of the stomach. No abnormalities were noted at necropsy of animals that were killed at the end of the study.

Any other information on results incl. tables

Mortality Data

Dose Level mg/kg

Sex

Number of Animals Treated

Deaths During Day of Dosing
(Hours)

Deaths During Period After Dosing
(Days)

Deaths

½

1

2

4

1

2

3

4

5

6

7

8-14

300

Female

3

0

0

0

0

0

0

0

0

0

0

0

0

0/3

Female

3

0

0

0

0

1

0

0

0

0

0

0

0

1/3

2000

Female

3

0

0

0

2*

1

 

 

 

 

 

 

 

3/3

*=    Animals 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 Clinical Observations - 300 mg/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

1-1

Female

0

0

0

H

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-2

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

X

 

 

 

 

 

 

 

 

 

 

 

 

 

3-2

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

H =  Hunched posture

X =  Animal dead

Individual Clinical Observations - 2000 mg/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

AL

HA

HA

X

 

 

 

 

 

 

 

 

 

 

 

 

 

2-1

Female

0

0

PrRl
RdTo

PrRlRd
HoEX
*

 

 

 

 

 

 

 

 

 

 

 

 

 

 

2-2

Female

0

0

PrRl
RdTo

PrRlRd
HoEX
*

 

 

 

 

 

 

 

 

 

 

 

 

 

 

0=   No signs of systemic toxicity

A =  Ataxia

L =  Lethargy

H =  Hunched posture

Pr = Prostration

Rl = Labored respiration

Rd =Decreased respiratory rate

To =Occasional body tremors

Ho =      Hypothermia

E =  Pallor of the extremities

X =  Animal dead

*=    Animals 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 Body Weight Changes - 300 mg/kg

Dose Level

mg/kg

Animal Number and Sex

Body Weight (g) at Day

Body Weight (g)
at Death

Body Weight Gain (g) During Week

0

7

14

1

2

300

1-0 Female

152

174

186

 

22

12

1-1 Female

160

178

192

 

18

14

1-2 Female

156

170

187

 

14

17

3-0 Female

177

194

212

 

17

18

3-1 Female

168

-

-

158

-

-

3-2 Female

166

187

200

 

21

13

- = Animal dead

Individual Body Weights and Body Weight Changes - 2000 mg/kg

Dose Level

mg/kg

Animal Number and Sex

Body Weight (g) at Day

Body Weight (g)
at Death

Body Weight Gain (g) During Week

0

7

14

1

2

2000

2-0 Female

161

-

-

154

-

-

2-1 Female

164

-

-

158

-

-

2-2 Female

158

-

-

156

-

-

-=    Animal dead

Individual 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

1-1 Female

Killed Day 14

No abnormalities detected

1-2 Female

Killed Day 14

No abnormalities detected

3-0 Female

Killed Day 14

No abnormalities detected

3-1 Female

Found dead Day 1

Liver: patchy pallor

Kidneys: dark

Gastric mucosa: epithelial sloughing

Non-glandular epithelium of the stomach: hemorrhagic

2-2 Female

Killed Day 14

No abnormalities detected

 

Individual Necropsy Findings - 2000 mg/kg

Dose Level
mg/kg

Animal Number
and Sex

Time of Death

Macroscopic Observations

2000

2-0 Female

Found dead Day 1

Liver: patchy pallor

Kidneys: pale

Gastric mucosa:hemorrhagic

                              epithelial sloughing

2-1 Female

Humanely killed Day 0

Liver: pale

Kidneys: pale

Stomach: clear liquid present

Gastric mucosa: epithelial sloughing

2-2 Female

Humanely killed Day 0

Liver: pale

Kidneys: pale

Stomach: clear liquid present

Gastric mucosa: epithelial sloughing


 

Applicant's summary and conclusion

Interpretation of results:
Toxicity Category IV
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 approximately 500 mg/kg body weight (Globally Harmonized Classification System – Category 4, >300 - 2000 mg/kg body weight and is also according to the EU C&L rules).

The test item was also classified as Category 4 according to Regulation (EC) No. 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures. It is reasonable to assume that the Signal Word “Warning” and the Hazard Statement “H302: Harmful if swallowed” are therefore required.
Executive summary:

The acute oral toxicity of the test substance, TM 11-213, was assessed according to OECD TG 423 using Wistar rats. At 300 mg/kg bw one animal died at the next higher dose of 2000 mg/kg bw all animals died, within the first day. At macroscopy pale liver and kidneys were observed and effects in the gastrointestinal tract indicating severe irritation: fluid in the stomach and sloughing in the epidermis. The toxicity of this substance is somewhat unexpected. Other hydrocarbon aldehydes of similar C-atoms have toxicity > 2000 mg/kg bw. When using the scheme in the guideline for this substance an LD50 of 500 mg/kg bw is derived.