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Description of key information

Acute oral toxicity. Key study:

The key study (1973) on oral acute toxicity was conducted in rats using alpha terpinene. 10 rats per treatment were exposed to the following doses: 1.05, 1.31, 1.64, 2.05 and 5 g/kg. Rats were observed for 14 days.

 

No clinical findings and deaths occurred in the lowest tested dose (1.05 g/kg). Lethargy was observed on the day of dosing in rats dosed at 1.31, 1.64, 2.05 and 5 g/kg. The rats dosed at 2.05 and 5 g/kg exhibited loss of righting reflex and piloerection.

 

The exposure to the middle doses (1.31 and 1.64 g/kg) caused 40% of death. 80% of death animals were found at 2.05 g/kg within 10 days after a single exposure. The highest dose caused 100% of death after 24 hrs. (8 deaths) and 48 hrs (2 deaths), respectively.

 

Based on the results the LD50 was calculated to be 1.68 g/kg (1.46 - 1.90).

Acute inhalation toxicity. Data waiving (study scientifically not necessary):

According to REACH Annex VIII, column 2: In addition to the oral route, for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. The choice for the second route will depend on the nature of the substance and the likely route of human exposure. If there is only one route of exposure, information for only that route needs to be provided. The information is provided for dermal route.

Acute dermal toxicity: Key study:

Test method according to OECD 402, GLP study. The LD50 of the test item is higher than 2000 mg/kg body weight by dermal route in rats.

3 female Sprague-Dawley rats were tested for a exposure period of 24 h. Initially, a range finding study was conducted at a single dose of 2000 mg/kg bw in one animal. Based on the results of range finding two additional rats were tested at a dose of 2000 mg/kg bw. No mortality was observed. The body weight evolution of the animals remained normal during the study. Erythema was noted in treated animals (3/3) between days 1 and 7. This reaction was associated with dryness of the skin (3/3) between days 3 and 12 and scab (3/3) between days 3 and 8. The skin recovered a normal aspect at day 13. No other clinical signs were observed. The macroscopic examination of the animals at the end of the study did not reveal treatment related effects.

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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:
1973
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
not specified
GLP compliance:
not specified
Test type:
standard acute method
Limit test:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Alpha Terpinene, 72-227
- Purity test date: not specified
- Type: Constituent

Species:
rat
Strain:
not specified
Sex:
not specified
Details on test animals or test system and environmental conditions:
TEST ANIMALS - rats
Route of administration:
oral: unspecified
Vehicle:
not specified
Details on oral exposure:
VEHICLE - not specified
Doses:
5 doses tested: 1.05, 1.31, 1.64, 2.05 and 5 g/kg
No. of animals per sex per dose:
10 animals per dose
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Necropsy of survivors performed: not specifed
- Other examinations performed: clinical signs
Statistics:
LD50 calculation
Key result
Sex:
not specified
Dose descriptor:
LD50
Effect level:
ca. 1.68 other: g/kg
Based on:
not specified
95% CL:
ca. 1.46 - ca. 1.9
Mortality:
Mortality was observed on day 1 at 1.31, 1.64, 2.05 and 5 g/kg in four, one, three and eight rats, respectively.
On day 2, the deaths were observed at dose levels: 1.64 g/kg (two animals) and at 2.05 g/kg (four animals) as well as at dose 5 g/kg (remaining two rats)
In additon, on day 4, one rat was found dead (dose level: 1.64 g/kg). Also, on day 10 there was a mortality observed in one rat at dose level of 2.05 g/kg.
Clinical signs:
The clinical signs depended on the dose tested

1.05 g/kg - no clinical signs
1.31 g/kg - lethargy
1.64 g/kg - lethargy
2.05 g/kg - lethargy, loss of righting reflex, piloerection
5 g/kg - lethargy, loss of righting reflex, piloerection

Body weight:
not specified
Gross pathology:
not specified

Table 1. Distribution of mortality

Group number

Dose Level

(g/Kg)

Deaths/ number of animals

Observation days

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1

1.05

0/10

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2

1.31

4/10

4

0

0

0

0

0

0

0

0

0

0

0

0

0

3

1.64

4/10

1

2

0

1

0

0

0

0

0

0

0

0

0

0

4

2.05

8/10

3

4

0

0

0

0

0

0

0

1

0

0

0

0

5

5

10/10

8

2

0

0

0

0

0

0

0

0

0

0

0

0

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
Based on the reults the LD50 was claculated to be 1.68 g/kg (1.46 - 1.90). Alpha terpinene is classifed as oral acute tox, category 4 accroding to GHS/EU CLP.
Executive summary:

In this study, an on oral acute toxicity was conducted in the rat using alpha terpinene.

 

Ten rats per treatment were exposed to the following doses: 1.05, 1.31, 1.64, 2.05 and 5 g/kg. Rats were observed for 14 days.

 

No clinical findings and deaths occurred in the lowest tested dose (1.05 g/kg). Lethargy was observed on the day of dosing in rats dosed at 1.31, 1.64, 2.05 and 5 g/kg. The rats dosed at 2.05 and 5 g/kg exhibited loss of righting reflex and piloerection.

 

The exposure to the middle doses (1.31 and 1.64 g/kg) caused 40% of death. 80% of death animals were found at 2.05 g/kg within 10 days after a single exposure.

 

The highest dose caused 100% of death after 24 hrs. (8 deaths) and 48 hrs (2 deaths), respectively.

 

Based on the results the LD50 was calculated to be 1.68 g/kg (1.46 - 1.90).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 680 mg/kg bw
Quality of whole database:
Peer reviewed publication with Klimisch=2.

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:
05 November 2019 - 21 November 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
A relative humidity lower than 30% was registered on 19 and 20 November 2019. The minimum value measured was 25%. As no effect was noted on the health of the animals, this deviation is considered as without impact on the conclusion of the study.
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Remarks:
(SPF Caw)
Sex:
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: yes
- Age at study initiation: 8 weeks
- Weight at study initiation: 207.7 g (SD = 6.7 g)
- Fasting period before study: not specified.
- Housing: During the treatment, the animals were kept in individual cages. On D1, the animals were put together into their cage. The rats were kept in solid-bottomed clear polycarbonate cages with a stainless steel mesh lid. Each cage contains dust free wood shavings which were changed at least 2 times a week. Each cage was installed in conventional air conditioned animal husbandry.
- Diet (e.g. ad libitum): Ad libitum. Teklad Global 16% Protein Rodent Diet (ENVIGO 2016).
- Water (e.g. ad libitum): Ad libitum. Drinking water (tap-water from public distribution system). Microbiological and chemical analyses of the water were carried out once every six months by Bureau Veritas – Eurofins (FRANCE).
- Acclimation period: the animals were acclimatized for at least 5 days before treatment.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19ºC to 25ºC
- Humidity (%): 30 to 70%
- Air changes (per hr): ≥10 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light (07.00 to 19.00) and 12 hours dark cycle.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage: at least 10% of the body surface area.
- Type of wrap if used: 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).

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg body weight
- Concentration (if solution): N/A
- Constant volume or concentration used: 2.43 mL/kg body weight (based on 0.835 relative density and 98.5% purity)
- For solids, paste formed: N/A
Duration of exposure:
24 h
Doses:
Range finding study: 2000 mg/kg body weight
Main study: 2000 mg/kg body weight
No. of animals per sex per dose:
Range finding study: 1 female per dose
Main study: 2 females per dose
Control animals:
yes
Remarks:
(study performed on three females receiving distilled water by topical application under requirements of OECD Guideline 402)
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: At each step, the animals were observed three times on test day 0 (day of administration), i.e. at T0+30 min, T0+3h and T0+5h, and once daily during days 1 to 14 post administration. The body weights were recorded on test day 0 (just before administration) then on D2, D7, and D14.
- Necropsy of survivors performed: yes. At termination, gross pathological findings were recorded and reported.
- Other examinations performed:
Clinical observations: spontaneous activity, Preyer’s reflex (noise), respiratory rate, convulsions, tremors, body temperature, muscle tone, palpebral opening, pupil appearance, salivation, lachrymation, righting reflex and treatment site (erythema, dryness of the skin, scab, etc.)
Key result
Sex:
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 treated animals (3/3) between Days 1 and 7. This reaction was associated with dryness of the skin (3/3) between Days 3 and 12 and scab (3/3) between Days 3 and 8. The skin recovered a normal aspect at Day 13.
Body weight:
The body weight evolution of the animals remained normal throughout the study.
Gross pathology:
The macroscopic examination of the animal at the end of the study did not reveal treatment-related changes.

Table 1: Body weight and weight gain in grams

Females D0 D2 D2-D0 D7 D7-D0 D14 D14-D0
Rf4434 212 205 -7 218 6 232 20
Rf4435 200 200 0 210 10 220 20
Rf4436 211 207 -4 226 15 237 26
MEAN 207.7 204.0 -3.7 218.0 10.3 229.7 22.0
S.D. 6.7 3.6 3.5 8.0 4.5 8.7 3.5

Interpretation of results:
other: No category (CLP Regulation EC no. 1272/2008)
Conclusions:
The LD50 of the test item is higher than 2000 mg/kg body weight by dermal route in rats.

Executive summary:

The acute dermal toxicity of the test item was studied according to OECD Guideline 402 (GLP study). 3 female Sprague-Dawley rats were tested for a exposure period of 24 h. Initially, a range finding study was conducted at a single dose of 2000 mg/kg bw in one animal. Based on the results of range finding two additional rats were tested at a dose of 2000 mg/kg bw. No mortality was observed. The body weight evolution of the animals remained normal during the study. Erythema was noted in treated animals (3/3) between days 1 and 7. This reaction was associated with dryness of the skin (3/3) between days 3 and 12 and scab (3/3) between days 3 and 8. The skin recovered a normal aspect at day 13. No other clinical signs were observed. The macroscopic examination of the animals at the end of the study did not reveal treatment related effects. Based on these results, the test item was found to be non toxic, with an LD50 > 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Study report with Klimisch=1.

Additional information

Justification for classification or non-classification

Based on the available results, alpha terpinene is classifed as oral acute tox, category 4 according to EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.