Reaction mass of (4R)-isopropenyl-1-methylcyclohexene and (4S)-isopropenyl-1-methylcyclohexene and 1-isopropyl-4-methylcyclohexa-1,3-diene and 1-isopropyl-4-methylcyclohexa-1,4-diene and 4-isopropylidene-1-methylcyclohexene

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 May 2016 - 31 May 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

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 2

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 3

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

Constituent 4

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

Constituent 5

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 1

Reference substance name:

(3R)-isopropylidene-6-methylcyclohexene

Cas Number:

43124-56-7

Molecular formula:

C10H16

IUPAC Name:

(3R)-isopropylidene-6-methylcyclohexene

impurity 2

Reference substance name:

(3S)-isopropylidene-6-methylcyclohexene

Cas Number:

62504-16-9

Molecular formula:

C10H16

IUPAC Name:

(3S)-isopropylidene-6-methylcyclohexene

impurity 3

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 4

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 5

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 6

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 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

Test material form:

liquid

Details on test material:

Batch No.: 173391
Purity: 85.8% (sum of the five main constituents)
Name of test material (as cited in study report): Reaction mass of dl-limonene, alpha- gamma-terpinenes, terpinolene
Physical state: colourless - slightly yellow liquid
Storage conditions: +2°C to +8°C, under nitrogen and protected from light
Expiry date: 1 October 2016

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage JANVIER LABS (53940 Le Genest St Isle – France),
- Females nulliparous and non-pregnant
- Age at study initiation: 7 weeks old (male) and 8 weeks old (female)
- Weight at study initiation: for male from 260 to 280 g, and for female from 204 to 221 g
- Housing: individually in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid.
- Diet ad libitum (SAFE - A04)
- Water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): 10 at least
- Photoperiod (hrs dark / hrs light): 12 hours light (07.00 to 19.00)

Administration / exposure

Type of coverage:

other: non occlusive porous gauze dressing

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

Animals received by topical application, under non occlusive porous gauze dressing (50 mm x 50 mm non-woven swab of 4-layer patch from MEDISTOCK) secured in position with a strip of surgical adhesive tape (50 mm wides hypoallergenic microporeTM adhesive tape from 3M), an effective dose of 2000 mg/kg body weight administered under a volume of 2.36 mL/kg body weight (according to the calculated density), during 24 hours. After 24-hour exposure period, the gauze dressings were removed and the treated areas were rinsed with distilled water.

Duration of exposure:

24 hours

Doses:

2000 mg/kg equivalent to 2.36 mL/kg bw

No. of animals per sex per dose:

5

Control animals:

other: historical control

Details on study design:

Systematic examinations are carried out to identify any behavioral or toxic effects on the major physiological functions 1 hour, 3 hours, 5 hours, 24 hours and 48 hours after administration of the test item and continued on the following days if necessary.
This examination focusses particularly on a list of symptoms, recorded as "present" or "absent" on the observation sheet.
These observations are compared to control data.
The integrity of the skin on the application site is noted.
Observations and a mortality report willare then be carried out every day for 14 days. The time of death is recorded individually in days and hours (if possible).
Weight changes in animals from treated group will beare compared to those of control animals.
Necropsies are carried out on animals that died during the test. Macroscopic observations are recorded on individual autopsy sheets.

Results and discussion

Effect levels

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study.

Clinical signs:

No systemic clinical signs related to the administration of the test item were observed.
Erythema was noted in all animals at 24-hour post-dose and was totally reversible on day 11. Dryness of the skin and scab were noted between days 6 and 10.

Body weight:

The body weight evolution of the animals remained normal throughout the study.

Gross pathology:

The macroscopic examination of the animals at the end of the study did not reveal any treatment-related changes.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The LD50 of test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE is higher than 2000 mg/kg body weight by dermal route in the rat.
Test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE does not have to be classified in accordance with Regulation EC No. 1272/2008 on classification, labelling and packaging of substances and mixtures. No signal word or hazard statement is required.

Executive summary:

REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE was applied onto the intact skin of 10 Sprague Dawley rats (5 males and 5 females) at the single dose of 2000 mg/kg body weight. The experimental protocol was established on the basis ohe official method as defined in OECD Guideline No. 402 dated February 24^th, 1987 and test method B.3 of Council regulation No. 440/2008 of 30 May 2008.

No mortality occurred during the study. No systemic clinical signs related to the administration of the test item were observed. Erythema was noted in all animals at 24-hour post-dose and was totally reversible on day 11. Dryness of the skin and scab were noted between days 6 and 10. Body weight gain of the animals remained normal throughout the study.

The macroscopic examination of the animals at the end of the study did not reveal any treatment-related changes.

In conclusion , the LD₅₀ of test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENEis higher than 2000 mg/kg body weight by dermal route in the rat.

 

Test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE does not have to be classified in accordance with Regulation EC No. 1272/2008 on classification, labelling and packaging of substances and mixtures.

No signal word or hazard statement is required.