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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May-June 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose:
read-across: supporting information

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
23 October 2015
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Test material form:
liquid
Details on test material:
Batch No. : FAB-01072015
Purity : 96.1% (sum of the two main constituents)
Name of test material (as cited in study report): alpha-pinene multiconstituent
Physical state: colourless liquid
Storage Conditions: +2°C to +8°C, under nitrogen and protected from light
Expiry Date: 30 June 2016

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS:
Sprague Dawley rats supplied by Elevage JANVIER LABS (53940 Le Genest St Isle – France)
Age at study initiate: 8 or 9 weeks old
Weight at study initiation: 185-230g
Housing: Animals were housed by group of three in solid bottomed clear polycarbonate cages with stainless steel mesh lid.
Fasting period before study: Food was removed on Day 1 and then redistributed 4 h after administration of the test item.
Diet: Foodstuff (SAFE, A04), ad libitum
Water: Drinking water (tap water from public distribution system), ad libitum
Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS:
Air recycling: at least 10 cycles per hour,
Temperature: 22° C ± 3° C,
Relative humidity: from 30 % to 70 %,
Lighting: circadian cycle (12 hrs day / 12 hrs darkness).

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: for 2000 mg/kg dose no vehicle, for 300 mg/kg vehicle is olive oil
Details on oral exposure:
In the first step of the study, the test item was administered by gavage under a volume of 2.33 mL/kg body weight (corresponding to 2000 mg/kg, according to the calculated relative density).

In the second and the third step of the study, 0.35 mL of the test item (corresponding to 300 mg/kg) was added to 1.98 mL of olive oil.
Doses:
2000 mg/kg and 300 mg/kg
No. of animals per sex per dose:
3 female rats for 2000 mg/kg
6 female rats at 300 mg/kg
Control animals:
other: historical data: Study No. TAO423-2016-002
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:
> 500 mg/kg bw
Based on:
test mat.
Mortality:
Three mortalities were noted in animals treated at the dose of 2000 mg/kg body weight, at 24 hours post-dose.

No mortality was noted in the animals treated at the dose of 300 mg/kg body weight.
Clinical signs:
At the dose of 2000 mg/kg the mortalities were preceded by a decrease in spontaneous activity (1/3) at 30 minutes post-dose.
At the dose of 300 mg/kg no clinical signs related to the administration of the test item were observed during the study.
Body weight:
The body weight evolution of the animals remained normal throughout the study at the dose of 300 mg/kg.
No data are available at the dose of 2000 mg/kg (animal found animals found dead at T0 + 21 hours and 19 minutes).
Gross pathology:
At the dose of 200 mg/kg the macroscopic examination of the animals revealed a thinning of the corpus (2/3).
At the dose of 300 mg/kg the macroscopic examination of the animals at the end of the study did not reveal treatment related changes.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The LD50 of test item alpha-pinene multiconstituent is higher than 300 mg/kg body weight and lower than 2000 mg/kg by oral route in the rat.
In accordance with guideline OECD 423, the LD50 cut-off of the test item may be considered as 500 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 alpha-pinene multiconstituent has to be classified in category 4. The signal word “Warning” and hazard statement H302 “Harmful if swallowed” are required
Executive summary:

The test item alpha-pinene multiconstituent was administered to a group of 3 female Sprague Dawley rats at the dose of 2000 mg/kg body weight and then to a group of 6 female Sprague Dawley rats at the dose of 300 mg/kg body weight. The experimental protocol was established according to the official method as defined in the O.E.C.D. Test Guideline No. 423 dated December 17th, 2001 and the test method B.1tris of the Council regulation No. 440/2008.

Three mortalities were noted in animals treated at the dose of 2000 mg/kg body weight, at 24 hours post-dose.

The mortalities were preceded by a decrease in spontaneous activity (1/3) at 30 minutes post-dose. No other signs of systemic toxicity were noted.

Rigor mortis were noted before the necropsy in two animals (2/3). The macroscopic examination of the animals revealed a thinningof the corpus (2/3).

 

No mortality was noted in the animals treated at the dose of 300 mg/kg body weight.

No clinical signs related to the administration of the test item were observed during the study.

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

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

In conclusion, the LD50 of test item alpha-pinene multiconstituent is higher than 300 mg/kg body weight and lower than 2000 mg/kg by oral route in the rat.

In accordance with guideline OECD 423, the LD50 cut-off of the test item may be considered as 500 mg/kg body weight by oral route in the rat.