Reaction mass of 3-isopropyl-6-methylenecyclohexene and (4R)-1-methyl-4-(prop-1-en-2-yl)cyclohexene and (4S)-1-methyl-4-(prop-1-en-2-yl)cyclohexene

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 11 to 26, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Well conducted and well described study in accordance with GLP and OECD guideline 423 without any deviation.

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable

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:

(S)-p-mentha-1,8-diene

EC Number:

227-815-6

EC Name:

(S)-p-mentha-1,8-diene

Cas Number:

5989-54-8

Molecular formula:

C10H16

IUPAC Name:

(4S)-isopropenyl-1-methylcyclohexene

Constituent 2

Reference substance name:

(R)-p-mentha-1,8-diene

EC Number:

227-813-5

EC Name:

(R)-p-mentha-1,8-diene

Cas Number:

5989-27-5

Molecular formula:

C10H16

IUPAC Name:

(4R)-isopropenyl-1-methylcyclohexene

Constituent 3

Reference substance name:

p-mentha-1(7),2-diene

EC Number:

209-081-9

EC Name:

p-mentha-1(7),2-diene

Cas Number:

555-10-2

Molecular formula:

C10H16

IUPAC Name:

3-isopropyl-6-methylenecyclohexene

impurity 1

Reference substance name:

p-mentha-1,3-diene

EC Number:

202-795-1

EC Name:

p-mentha-1,3-diene

Cas Number:

99-86-5

Molecular formula:

C10H16

IUPAC Name:

1-isopropyl-4-methylcyclohexa-1,3-diene

impurity 2

Reference substance name:

p-mentha-1,4-diene

EC Number:

202-794-6

EC Name:

p-mentha-1,4-diene

Cas Number:

99-85-4

Molecular formula:

C10H16

IUPAC Name:

1-isopropyl-4-methylcyclohexa-1,4-diene

impurity 3

Reference substance name:

p-mentha-1,4(8)-diene

EC Number:

209-578-0

EC Name:

p-mentha-1,4(8)-diene

Cas Number:

586-62-9

Molecular formula:

C10H16

IUPAC Name:

4-isopropylidene-1-methylcyclohexene

impurity 4

Reference substance name:

p-cymene

EC Number:

202-796-7

EC Name:

p-cymene

Cas Number:

99-87-6

Molecular formula:

C10H14

IUPAC Name:

1-isopropyl-4-methylbenzene

impurity 5

Reference substance name:

(1R,6S)-3,7,7-trimethylbicyclo[4.1.0]hept-3-ene

Cas Number:

20296-50-8

Molecular formula:

C10H16

IUPAC Name:

(1R,6S)-3,7,7-trimethylbicyclo[4.1.0]hept-3-ene

impurity 6

Reference substance name:

(1S)-3,7,7-trimethylbicyclo[4.1.0]hept-3-ene

EC Number:

207-856-6

EC Name:

(1S)-3,7,7-trimethylbicyclo[4.1.0]hept-3-ene

Cas Number:

498-15-7

Molecular formula:

C10H16

IUPAC Name:

(1S,6R)-3,7,7-trimethylbicyclo[4.1.0]hept-3-ene

impurity 7

Reference substance name:

(R)-5-isopropyl-2-methylcyclohexa-1,3-diene

EC Number:

224-167-6

EC Name:

(R)-5-isopropyl-2-methylcyclohexa-1,3-diene

Cas Number:

4221-98-1

Molecular formula:

C10H16

IUPAC Name:

(5R)-isopropyl-2-methylcyclohexa-1,3-diene

impurity 8

Reference substance name:

(5S)-isopropyl-2-methylcyclohexa-1,3-diene

Cas Number:

2243-33-6

Molecular formula:

C10H16

IUPAC Name:

(5S)-isopropyl-2-methylcyclohexa-1,3-diene

impurity 9

Reference substance name:

(1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane

EC Number:

227-337-8

EC Name:

(1S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane

Cas Number:

5794-04-7

Molecular formula:

C10H16

IUPAC Name:

(1S,4R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane

impurity 10

Reference substance name:

(1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane

EC Number:

227-336-2

EC Name:

(1R)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane

Cas Number:

5794-03-6

Molecular formula:

C10H16

IUPAC Name:

(1R,4S)-2,2-dimethyl-3-methylenebicyclo[2.2.1]heptane

impurity 11

Reference substance name:

(-)-pin-2(3)-ene

EC Number:

232-077-3

EC Name:

(-)-pin-2(3)-ene

Cas Number:

7785-26-4

Molecular formula:

C10H16

IUPAC Name:

(1S,5S)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

impurity 12

Reference substance name:

(+)-pin-2(3)-ene

EC Number:

232-087-8

EC Name:

(+)-pin-2(3)-ene

Cas Number:

7785-70-8

Molecular formula:

C10H16

IUPAC Name:

(1R,5R)-2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

impurity 13

Reference substance name:

7,7-dimethyl-2-methylene-, (1S,4R)-bicyclo[2.2.1]heptane

Cas Number:

7378-37-2

Molecular formula:

C10H16

IUPAC Name:

7,7-dimethyl-2-methylene-, (1S,4R)-bicyclo[2.2.1]heptane

impurity 14

Reference substance name:

7,7-dimethyl-2-methylene-(1R,4S)-bicyclo[2.2.1]heptane

Cas Number:

116724-26-6

Molecular formula:

C10H16

IUPAC Name:

7,7-dimethyl-2-methylene-(1R,4S)-bicyclo[2.2.1]heptane

impurity 15

Reference substance name:

Non identified impurities

Molecular formula:

Not applicable

IUPAC Name:

Non identified impurities

Test material form:

liquid

Details on test material:

Batch No.: ED50F50R
Purity: 69% (sum of the three main constituents)
Name of test material (as cited in study report): Dipentene multiconstituent
Physical state: colourless liquid
Storage conditions: +2°C to +8°C, under nitrogen and protected from light

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER (53940 Le Genest St Isle-France)
- Age at study initiation: 9 weeks
- Weight at study initiation: 207-230 g
- Fasting period before study: 24 hours
- Housing: In group of three animals in solid bottomed clear polycarbonate cages with sawdust bedding
- Diet: M20-SDS; ad libitum
- Water: Tap-water from public distribution system; ad libitum
- Acclimation period: Five days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 ° C
- Humidity: 30-70%
- Air change: 15 changes/hour
- Photoperiod: 12 hours dark/12 hours light

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 2.28 mL/kg bw


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Dose was selected as per specified in the guidelines

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

Six females

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observed every day, weighed on Day 0, 2, 7 and 14
- Necropsy of survivors performed: yes; animals were anaesthetised with sodium phenobarbital on Day 14 for necropsy
- Other examinations performed: clinical signs

Statistics:

No data

Results and discussion

Preliminary study:

Not applicable

Effect levels

Effect levels 1

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Effect levels 2

Sex:

female

Dose descriptor:

other: LD50 cut-off

Effect level:

ca. 2 500 mg/kg bw

Based on:

test mat.

Mortality:

1/6 on Day 6

Clinical signs:

other: - Decrease in spontaneous activity (4/6), muscle tone and righting reflex (3/6); increased salivation and piloerection (3/6) on Day 1; animals recovered to normal at 24 hours post dose.

Gross pathology:

- White thinning of the corpus of the animal that died on Day 6
- Thickness (5/5) and white coloration (2/5) of forestomach

Other findings:

None

Any other information on results incl. tables

Table 1: Body weight and weight gain in grams

Animals	Day 0	Day 2	Day 2-Day 0	Day 7	Day 7-Day 0	Day 14	Day 14-Day 0
1	213	198	-15	Dead	Dead	Dead	Dead
2	230	237	7	256	26	285	55
3	207	211	4	245	38	249	42
4	219	225	6	241	22	264	45
5	221	222	1	252	31	268	47
6	225	239	14	263	38	272	47
Mean	219.2	222	2.8	251.4	31	267.2	47.2
Standard deviation	8.3	15.6	9.7	8.7	7.1	13	4.8

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The oral LD50 for dipentene multiconstituent was found to be greater than 2000 mg/kg bw in the Sprague-Dawley rats and therefore it is not classified according to Directive 67/548/EEC and CLP Regulation (EC) N° 1272/2008.

Executive summary:

In an oral acute toxicity study (limit test) performed in accordance with GLP and OECD guideline 423, groups of six Sprague-Dawley female rats received a single oral (gavage) dose of dipentene multiconstituent at 2000 mg/kg bw. Animals were then observed for mortality, body weight and clinical signs of toxicity for 14 days and were all macroscopically necropsied after sacrifice. The observations were compared to historical control data.

One animal died on Day 6 during the study. A decrease in spontaneous activity (4/6), muscle tone and righting reflex (3/6) and increased salivation and piloerection (3/6) was noted on first day of study. All animals recovered to normal behavior at 24 hours post dose. A 7% decrease in body weight on Day 2 was observed in animal that died on Day 6. In surviving animals absence of body weight gain was noted on Day 2 that recovered normal body weight on Day 7. Thickness (5/5) and white coloration (2/5) of forestomach of surviving animals and white thinning of the corpus of the dead animal was noted on necropsy. The oral LD50 was found to be greater than 2000 mg/kg bw in rats.

In accordance with OECD guideline No 423, the LD50 cut-off of the test item may be considered as 2500 mg/kg bw by oral route in the rat.

The oral LD50 for dipentene multiconstituent was found to be greater than 2000 mg/kg bw in the Sprague-Dawley rats and therefore it is not classified according to Directive 67/548/EEC and CLP Regulation (EC) N° 1272/2008.