Cyclohexane, 1-methyl-4-(1-methylethyl)-, tetradehydro deriv.

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to GLP and valid methods, therefore it is considered relevant, reliable and adequate for classification.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 2

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

Test material information

Constituent 1

Reference substance name:

Reaction mass of 1-Methyl-4-(1-methylethenyl)cyclohexene and 1-Methyl-4-(1-methylethylidene)-cyclohexene and 1-methyl-4-(propan-2-yl)cyclohexa-1,3-diene

EC Number:

939-409-2

Cas Number:

1329-99-3

Molecular formula:

C10H16

IUPAC Name:

Reaction mass of 1-Methyl-4-(1-methylethenyl)cyclohexene and 1-Methyl-4-(1-methylethylidene)-cyclohexene and 1-methyl-4-(propan-2-yl)cyclohexa-1,3-diene

Constituent 2

Reference substance name:

Isopropylmethylcyclohexane, tetradehydro derivative

EC Number:

215-532-0

EC Name:

Isopropylmethylcyclohexane, tetradehydro derivative

Cas Number:

1329-99-3

IUPAC Name:

1-isopropyl-4-methylcyclohexane

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): Depanol I; isopropylmethylcyclohexane, tetrahydro derivative; p-menthadiene;
- Substance type: Organic multi constituent
- Physical state: Liquid, colourless, characteristic odour
- Analytical purity: Terpene hydrocarbons: 98.9% (w/w); p- Menthadienes: 96.3%
- Impurities (identity and concentrations): See confidential details
- Composition of test material, percentage of components: See confidential details
- Purity test date: 2011-07-06
- Lot/batch No.: DEFM102174
- Expiration date of the lot/batch: 2013-06-25
- Stability under test conditions: Stability of the test item in the vehicle was established at 1.0 and 200 mg/mL concentration levels under Advinus study no.G8350.
- Storage condition of test material: Ambient (+18 to +36°C)
- Other: Density: 0.85 – 0.87 g/cm3; Water solubility: <0.5 g/L

Test animals

Species:

rat

Strain:

other: Wistar – HsdCpb: WU

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Toxicology, Department of Safety Assessment, Advinus Therapeutics Limited, Bangalore 560 058, India
- Age at study initiation: 9 to 10 weeks at treatment
- Weight at study initiation: 185.40 – 200.17 g
- Fasting period before study: Yes, 16-18 hours; water was not withheld. Food was offered 3-4 hours after dosing.
- Housing: Individually in standard polysulfone cages (Size: approximately L 425 x B 266 x H 175 mm), with stainless steel top grill having facilities for pelletted food and drinking water. Bedding: steam sterilized clean paddy husk was used and changed along with the cage twice a week.
- Diet (e.g. ad libitum): Teklad Certified (2014C) Global 14% Protein Rodent Maintenance Diet - Pellet (Certified) manufactured by Harlan Laboratories B.V. Maasheseweg 87c PO Box 553, 5800, AN Venray, The Netherland; ad libitum.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line water filter-cum-purifier manufactured by Eureka Forbes Ltd, Mumbai 400 001, India, in polycarbonate bottles with stainless steel sipper tubes; ad libitum.
- Acclimation period: From 5 to 7 days under laboratory condition after physical examination

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23°C
- Humidity (%): 58-67%
- Air changes (per hr): 12-15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: August 9, 2012 To: August 30, 2012

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle (if gavage):
- Justification for choice of vehicle: A quantity of 1.0 g of the test item was mixed with Milli-Q water and the volume made up to 5 mL. The test item was not miscible.
Hence a quantity of 1 g of test item was mixed with corn oil and the volume was made up to 5 mL to get the test item concentration of 200 mg/mL suspension. The test item formed a visibly homogenous suspension. Hence, corn oil was used as the vehicle to prepare the test item suspension.
- Lot/batch no. (if required): MKBG9425V

DOSAGE PREPARATION (if unusual): The test item suspension was prepared on each day of the test item administration. A quantity of 4.0 g of the test item was mixed by adding small quantity of the selected vehicle (corn oil) and the volume made up to 20 mL to get the test item concentration of 200 mg/mL suspension. Homogeneity of the test item in the vehicle was maintained by mixing, using glass rod, prior to treatment. Preparation was made prior (within 1 hour) to dosing.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: As per the Material Safety Data Sheet provided by the Sponsor, the Acute Oral toxicity (mg/kg) in mouse is >2000 mg/kg body weight, hence the test was started as per Annex 2d of the OECD 423. The starting dose was 2000 mg/kg body weight with 3 animals /step.

Doses:

2000mg/kg bw

No. of animals per sex per dose:

3 females per treatment step

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations: 5x on test day 1 and 1x daily during days 2-15 post administration; Weighing: on test day 1 (pre-administration), day 8 (7 days post administration) and day 15 (14 days post administration).
- Necropsy of survivors performed: yes

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: as per LD50 cut-off value

Mortality:

No pre-terminal deaths.

Clinical signs:

other: No clinical signs.

Gross pathology:

No abnormality was detected at necropsy in control and treatment groups.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Depanol I has an oral LD50 of >2000 mg/kg body weight and does not have to be classified as per CLP Regulation.

Executive summary:

The acute oral toxicity study with Depanol I in Wistar rats was tested according to the acute toxic class method. The test item suspended in corn oil was administered as oral gavage to overnight fasted (16 – 18 hours) 3 female rats at the dose of 2000 mg/kg body weight. Vehicle control group animals were administered corn oil. There were no toxic signs and pre-terminal deaths in treatment and control groups. Three additional female rats were tested at the same dose of 2000 mg/kg body weight. There were no toxic signs and pre-terminal deaths in treated and control groups. All animals of both groups gained body weight during 14 days observation period.

The rats of treatment and control groups were subjected to necropsy at termination and there were no abnormalities detected.

Based on the results of the present study, the test item, Depanol I has an oral LD50 of >2000 mg/kg body weight and does not have to be classified as per CLP Regulation.