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

Endpoint summary

Description of key information

Oral LD₅₀ was higer than 2000 mg/kg in an OECD 423 study conducted in rats in compliance with GLP.

Dermal LD₅₀ was higer than 2000 mg/kg in an OECD 402 study conducted in rats in compliance with GLP.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

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

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

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

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

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

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.

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

OECD 423 study without deviation, according to Good laboratory practice

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

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

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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)

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.

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.

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

OECD 402 study without deviation, according to Good laboratory practice

Justification for classification or non-classification

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.

Therefore, 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.

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.

Therefore, 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.