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

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

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 402 (Acute Dermal Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute 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%
- Composition of test material, percentage of components & impurities: See confidential details
- Purity test date: Letter of Composition 2011-7-6
- Lot/batch No.: DEFM102174
- Expiration date of the lot/batch: 2013-06-25
- Stability under test conditions: stable
- 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:

male/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: Males: 279.10 to 291.35 g; Females: 217.72 to 235.12 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 corn cob 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; water bottles were replenished once daily and the water bottles were changed once a week; 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

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsolateral thoracic surface of the skin (approx. 8 x 10 cm)
- % coverage: 10 % of body surface
- Type of wrap if used: Test item was applied directly to the clipped skin and covered with cotton gauze (size: Males: 9 x 6 cm; Females: 8 x 5 cm of 6 ply) and secured in position by adhesive tape wound around torso.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, initially with water and with Johnson’s Baby Soap (Johnson & Johnson U.S.A.© J&J*TM) and next with water. Washed animals were wiped dry with a cotton hand towel.
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000mg/kg bw ; 2.35 mL/kg bw
- Constant volume used: no

Duration of exposure:

24 hours

Doses:

2000 mg/kg bw equivalent to 2.35 mL/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing: Observations: 4x 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:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No pre-terminal deaths.

Clinical signs:

other: No clinical signs of toxicity, no local skin reaction.

Gross pathology:

There was no abnormality detected at necropsy.

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Based on the present study results, the acute dermal LD50 of Depanol I is greater than 2000 mg/kg bw in male and female Wistar rats.

Executive summary:

An acute dermal toxicity test was conducted with Wistar rats (male and female) to determine the potential for Depanol I to produce toxicity from a short term exposure via the dermal route.
Based on the individual body weight the undiluted test item at the dose of 2000 mg/kg [2.35 mL/kg based on the density of the test item 0.85 g/cm3] body weight was calculated and applied directly to the clipped skin of the animal to cover about 10% of body surface of the animal and covered with cotton gauze (size: Males: 9 x 6 cm; Females: 8 x 5 cm of 6 ply) and secured in position by adhesive tape wound around torso. The test item contact period with the skin was for 24 hours.
After the 24 hour contact period, the dressing was removed and the applied area was washed initially with water and with Johnson’s Baby Soap and again with water. Washed animals were wiped dry with a cotton hand towel. 
The treatment was initiated in 5 female rats at the dose of 2000 mg/kg body weight. There were no clinical signs of toxicity, local skin reaction or mortality observed. Hence 5 male rats were treated at the same dose of 2000 mg/kg body weight. All the rats were observed for clinical signs of toxicity, local skin reaction and mortality for 14 days post application. There were no clinical signs of toxicity, local skin reaction or mortality. At the end of observation period, all the animals were euthanized and subjected to necropsy. There were no abnormalities detected at the necropsy. Based on the results of the present study, the LD50 was above 2000 mg/kg bw and the test item does not have to be classified.