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: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 May 2016 - 01 June 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

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

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:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: by Elevage JANVIER LABS (53940 Le Genest St Isle – France),
- Females nulliparous and non-pregnant:
- Age at study initiation: 8 weeks
- Weight at study initiation: from 182 to 207 g
- Housing: animals were housed by group of three in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid.
- Diet ad libitum
- Water ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): from 19°C to 25°C
- Humidity (%): from 30% to 70%
- Air changes : at least ten changes per hour
- Photoperiod: twelve hours continuous light (07.00 to 19.00) and twelve hours darkness

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The animals of the treated group received an effective dose of 2000 mg/kg body weight of test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE.

In the first and the second step of the study, the test item was administered by gavage under a volume of 2.36 mL/kg body weight (corresponding to 2000 mg/kg according to the density of 0.846 g/mL supplied by the Sponsor) using a suitable graduated syringe fitted with an oesophageal metal canula

Doses:

2000 mg/kg corresponding to 2.36 mL/ kg bw

No. of animals per sex per dose:

6 (3 per step)

Control animals:

other: historical data

Details on study design:

Clinical observations and mortality were recorded every day for 14 days
Animals were weighed on day D0 (just before administering the test item) then on D2, D7, and D14.
On D14, the animals were anaesthetised with sodium pentobarbital and administration continued to fatal levels. Macroscopic observations were entered on individual autopsy sheets.
Only those organs likely to be modified in cases of acute toxicity were examined. No organ was removed and preserved in view to microscopic examinations.

Statistics:

none

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50 cut-off

Effect level:

2 500 mg/kg bw

Based on:

test mat.

Mortality:

Two mortalities were noted in the animals treated at the dose of 2000 mg/kg body weight, one at 48 hours post dose during the first step of the study and a one at 24 hours post dose during the second step of the study.

Clinical signs:

The mortalities were preceded by a decrease in spontaneous activity (2/2). No others clinical signs were noted. In the surviving animals (4/6), a decrease in spontaneous activity (4/4) was noted during the first hours of the test. The animals recovered a normal activity at 3-hour post dose. No others signs of systemic toxicity were noted.

Body weight:

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

Gross pathology:

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

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The LD50 of test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE is higher than 2000 mg/kg body weight by oral route in the rat. In accordance with OECD Guideline No. 423, the LD50 cut-off of the test item may be considered as 2500 mg/kg body weight by oral route in the rat.
According to the criteria for classification, packaging and labelling of dangerous substances and preparations in accordance with Regulation EC No. 1272/2008, test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE does not have to be classified.
According to GHS regulation, test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA- TERPINENES, TERPINOLENE is classified in category 5. Hazard statement H303 (may be harmful if swallowed) and signal word Attention are required.

Executive summary:

REACTION MASS OF DL-LIMONENE, ALPHA-GAMMA-TERPINENES, TERPINOLENE was administered to a group of 6 female Sprague Dawley rats at the dose of 2000 mg/kg body weight. The experimental protocol was established according to the official method as defined in OECD Guideline No. 423 dated December 17th, 2001 and test method B.1tris of Council regulation No. 440/2008.

 

Two mortalities were noted in the animals treated at the dose of 2000 mg/kg body weight, one at 48 hours post dose during the first step of the study and one at 24 hours post dose during the second step of the study. The mortalities were preceded by a decrease in spontaneous activity (2/2). No other clinical signs were noted. Rigor mortis was noted before the necropsy (2/2).The macroscopic examination of the animals revealed a thinning of forestomach and corpus with red spots (1/2).

 In the surviving animals (4/6), a decrease in spontaneous activity (4/4) was noted during the first hours of the test. The animals recovered a normal activity at a 3-hour post dose. No others signs of systemic toxicity were noted. The macroscopic examination of this animal 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, TERPINOLENE is higher than 2000 mg/kg body weight by oral route in the rat. In accordance with OECD Guideline No. 423, the LD₅₀ cut-off of the test item may be considered as 2500 mg/kg body weight by oral route in the rat.

 

According to the criteria for classification, packaging and labelling of dangerous substances and preparations in accordance with Regulation EC No. 1272/2008, test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA-TERPINENES, TERPINOLENE does not have to be classified.

No signal word or hazard statement is required.

According to GHS regulation, test item REACTION MASS OF DL-LIMONENE, ALPHA- GAMMA- TERPINENES, TERPINOLENE is classified in category 5. Hazard statement H303 (may be harmful if swallowed) and signal word Attention are required.