Registration Dossier

Administrative data

Description of key information

Acute oral toxicity (tested in a study similar to OECD TG 401): LD50 = 5000 mg/kg bw

Acute dermal toxicity (tested in a study similar to OECD TG 402): LD50 = 8000 mg/kg bw

Acute inhalation toxicity using route to route extrapolation from the oral route ca. 13000 mg/m3.

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:
1971
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The data considered sufficiently reliable for the purpose of hazard assessment
Reason / purpose for cross-reference:
reference to same study
Principles of method if other than guideline:
Standard acute method (limit test)
GLP compliance:
no
Remarks:
pre-GLP
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 113-150 g
- Fasting period before study: Animals were fasted overnight but provided water ad libitum.
- Housing: Animals were housed in groups of 5/sex in suspended stainless steel wire mesh cages.
- Diet: Food, ad libitum
- Water, ad libitum


Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw

Doses:
5 mL/kg bw
No. of animals per sex per dose:
5 animals/sex/dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for toxic symptoms and mortality at 0.5, 1, 2, 4 and 24 h, and then daily for 14 days after test item administration. Animals were weighed initially and at 14 days.
- Necropsy of survivors performed: Yes; gross pathological examination was performed for animals that died as well as animals surviving the 14 days observation period.
Statistics:
None
Preliminary study:
None
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
5 mL/kg bw
Based on:
test mat.
Remarks on result:
other: Four males were dead at 24 h and the remaining animals were lethargic and unthrifty in appearance.
Mortality:
- Four males were dead at 24 h and one male was sluggish and unthrifty in appearance.
Clinical signs:
- Females: Facial grooming, increased spontaneous activity and salivation were noted in all females at 0.5 h with one exhibiting an apparent equilibrium problem as evidenced by head tilting. Ataxia and slight CNS depression were noted at 2 h, and rigidity and sluggishness at 4 h. Unthriftiness and sluggishness were noted at 24 h. All females appeared normal at 48 h and thereafter.
- Males: Facial grooming, salivation and increased spontaneous activity were noted in two males at 0.5 and 1 h. Slight ataxia and slight CNS depression were noted in 3 males at 2 h. Occassional tremors in one male and withdrawl in one male were also noted at 2 h. All males exhibited rigidity and sluggishness at 4 h. The surviving male appeared normal at 48 h and thereafter.
Body weight:
- Mean body weight gain of the females was 50 g and the body weight of surviving male was 43 g.
Gross pathology:
- Gross pathological findings of the four males that died consisted of gastric hyperemia.
- No gross changes were noted in animals necropsied after 14 day observation period.
Other findings:
None

None

Interpretation of results:
other: not acute toxic
Remarks:
in accordance with EU CLP (EC 1272/2008 and its updates)
Conclusions:
Under the test conditions, the oral LD50 for Rosemary oil is 5000 mg/kg bw in rats. Based on this result, the substance is considered to be not acute toxic via the oral route.
Executive summary:

In an acute oral toxicity (limit test) study, a group of 5 rats/sex/dose were given a single oral dose of Rosemary oil at 5 mL/kg bw. Animals were then observed for mortality, clinical signs and body weight for 14 days and were all sacrificed for macroscopic examination.

Four males were dead at 24 h and one male was sluggish and unthrifty in appearance. Facial grooming, increased spontaneous activity and salivation were noted in all females at 0.5 h with one exhibiting an apparent equilibrium problem as evidenced by head tilting. Ataxia and slight CNS depression were noted at 2 h, and rigidity and sluggishness at 4 h. Unthriftiness and sluggishness were noted at 24 h. All females appeared normal at 48 h and thereafter. Facial grooming, salivation and increased spontaneous activity were noted in two males at 0.5 and 1 h. Slight ataxia and slight CNS depression were noted in 3 males at 2 h. Occassional tremors in one male and withdrawl in one male were also noted at 2 h. All males exhibited rigidity and sluggishness at 4 h. The surviving male appeared normal at 48 h and thereafter. Mean body weight gain of the females was 50 g and the body weight of surviving male was 43 g. Gross pathological findings of the four males that died consisted of gastric hyperemia. No gross changes were noted in animals necropsied after 14 day observation period. In this study, the oral LD50 of test item was 5 mL/kg bw in rats. Under the test conditions, the oral LD50 for Rosemary oil is 5000 mg/kg bw in rats.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Only basic data given in the available studies. However, the two studies showed consistent results. As a worst-case, the lowest LD50 value was selected as the key effect level for this endpoint.

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:
1971
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Reason / purpose for cross-reference:
reference to same study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Weight at study initiation:
females: 2.95 and 2.73 kg
males: 2.19 and 3.33 kg
- Housing: Animals were confined to a specially designed restrainer during exposure.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: The substance was applied on the back and flanks of the rabbits. The skin was clipped and intact (n=2) or clipped and abraded (n=2).
- Type of wrap if used: dental dam

REMOVAL OF TEST SUBSTANCE
- Washing (if done): sponged with warm tap water.
- Time after start of exposure: 24 h

TEST MATERIAL
- Amount(s) applied: 10 mL/kg
- Concentration: Undiluted

VEHICLE
not aplicable
Duration of exposure:
24 hours
Doses:
10 mL/kg
No. of animals per sex per dose:
2
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observation of gross signs of systemic toxicity of percutaneous absorption of the test material was made during the day following application and daily and thereafter for 14 days. Gross signs of dermal irritation were determined and scored following removal of the binders at 24 hours and thereafter for a total of 14 days. Individual body weights were recorded initially and at termination of the study.
- Necropsy of survivors performed: yes
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 10 mL/kg bw
Based on:
test mat.
Mortality:
One death occured (day 4).
Clinical signs:
Ataxia was noted in one rabbit for three days. On day 4 this rabbit died. No toxic symptoms were observed in the remaining rabbits.
Body weight:
Body weight gain of the three surviving animals averaged 0.3 kg.
Gross pathology:
No gross pathological findings were noted in any of the animals.
Other findings:
- Other observations:
Very slight erythema was noted in Rabbit No. 21 at 24 hours and became well defined at day seven and moderate by day 10 through day 14. Very slight edema noted at 24 hours. !nduration at the application site occurred on day seven followed by sloughing. Similarly, very slight erythema noted in rabbit no. 22 at 24 hours became well-defined on day seven followed by sloughing. Well defined erythema and slight edema noted at 24 hours in the abraded rabbits became moderate by two to six days with moderate edema noted in Rabbit No. 23 at five days. By day eight the skin became indurated followed by sloughing in Rabbit No. 23.
Interpretation of results:
other: not acute toxic
Remarks:
in accordance with EU CLP (EC 1272/2008 and its updates)
Conclusions:
The acute dermal toxicity test showed an LD50 of > 10 mL/kg bw corresponding to ca. 8000 mg/kg bw. Based on this result, the substance is considered to be not acute toxic via the dermal route.
Executive summary:

Rosemary oil was tested in an acute dermal toxicity study similar to OECD TG 402. Four New Zealand White rabbits were exposed to the test substance via dermal application. 2 males and 2 females were exposed to 10 mL/kg bw undiluted test substance under occlusive conditions for 24 hours. The substance was applied on the back and flanks of the rabbits. The skin was clipped and intact in two rabbits (one male and one female) and clipped and abraded in the other two animals. Skin responses consisted of very slight to welldefined erythema and slight to very slight edema at 24 hours to moderate erythema and induration with skin sloughing by 14 days. After an observation period of 14 days animals were necropsied. Ataxia was noted in one rabbit for three days (animal with abraded skin). On day 4 this rabbit died. No toxic symptoms were observed in the remaining rabbits. Body weight gain of the three surviving animals averaged 0.3 kg. No gross pathological findings were noted in any of the animals. The LD50 was determined to be higher than 10 mL/kg bw corresponding with ca. 8000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
8 000 mg/kg bw

Additional information

Acute toxicity via oral route

A key study was identified (Lynch, 1971, Rel. 2). In this acute oral toxicity (limit test) study, a group of 5 rats/sex were given a single oral dose of Rosemary oil at 5 mL/kg bw.

Four males were dead at 24 h and one male was sluggish and unthrifty in appearance. Facial grooming, increased spontaneous activity and salivation were noted in all females at 0.5 h with one exhibiting an apparent equilibrium problem as evidenced by head tilting. Ataxia and slight CNS depression were noted at 2 h, and rigidity and sluggishness at 4 h. Unthriftiness and sluggishness were noted at 24 h. All females appeared normal at 48 h and thereafter. Facial grooming, salivation and increased spontaneous activity were noted in two males at 0.5 and 1 h. Slight ataxia and slight CNS depression were noted in 3 males at 2 h. Occassional tremors in one male and withdrawl in one male were also noted at 2 h. All males exhibited rigidity and sluggishness at 4 h. The surviving male appeared normal at 48 h and thereafter. Mean body weight gain of the females was 50 g and the body weight of surviving male was 43 g. Gross pathological findings of the four males that died consisted of gastric hyperemia. No gross changes were noted in animals necropsied after 14 day observation period. In this study, the oral LD50 of test item was 5 mL/kg bw in rats. In the supporting study (von Scramlik, 1959, Rel.4), the LD50 in rats was 6600 mg/kg bw.

 

Acute toxicity via dermal route

Rosemary oil was tested in an acute dermal toxicity study similar to OECD TG 402. Four New Zealand White rabbits were exposed to the test substance via dermal application. 2 males and 2 females were exposed to 10 mL/kg bw undiluted test substance under occlusive conditions for 24 hours. The substance was applied on the back and flanks of the rabbits. The skin was clipped and intact in two rabbits (one male and one female) and clipped and abraded in the other two animals. After an observation period of 14 days animals were necropsied. Ataxia was noted in one rabbit for three days. On day 4 this rabbit died. No toxic symptoms were observed in the remaining rabbits. Body weight gain of the three surviving animals averaged 0.3 kg. No gross pathological findings were noted in any of the animals. The LD50 was determined to be higher than 10 mL/kg bw corresponding with ca. 8000 mg/kg bw (Lynch, 1971).

 

Acute toxicity via inhalation

The acute inhalation toxicity of Rosemary oil is predicted based on the acute oral toxicity in accordance with the ECHA CLP guidance document (2017, page 250, 3.1.3.3.5: 1 mg/kg bw = 0.0052 mg/l (5.2 mg/m3). This means that an LD50 of 5000 mg/kg bw * 0.0052 = 26 mg/L = 26000 mg/m3. The acute inhalation is predicted to be = 13000 mg/m3 for Rosemary oil (using 100% inhalation and 50% oral absorption and an LD50 oral of = 5000 mg/kg bw).

 

An LC50 of ca. 13000 mg/m3 for Rosemary oil would not result in a classification for acute toxicity via inhalation according to CLP. Furthermore, assuming that Camphor (H332), the only classified constituent of Rosemary oil, is always present at < 25%, Rosemary oil does not require classification for acute toxicity via inhalation according to the Regulation (EC) No. 1272/2008.

 

Specific target organ toxicity: single exposure (oral)

Classification STOT SE 2 is suggested for one of the constituents in Rosemary oil, Camphor. Human epidemiological data report clear signs of CNS effects in humans following oral administration. Limited information is available regarding dermal and inhalatory exposure however effects via these routes cannot be excluded. Based on epidemiological data, the disseminated REACH dossier and the IFRA classification, STOT SE 2 (target: CNS) was assigned to Camphor. In the acute oral toxicity study with 5000 mg/kg bw Rosemary oil reversible effects on the CNS were reported. These are likely caused by the constituent, Camphor. For Rosemary oil no other data regarding specific target organ toxicity are available.

 

Based on the available information, it is concluded that all Rosemary oil qualities having ≥ 10% of Camphor should be classified as: STOT SE 2 (H371: May cause damage to CNS following exposure) according to the Regulation (EC) No. 1272/2008.

Justification for classification or non-classification

Based on the available data, the substance does not need to be classified for acute oral, dermal and inhalation toxicity in accordance with the criteria outlined in EU CLP (EC no 1272/2008 and its amendments). Based on the boundary composition and the presence of camphor in the substance, Rosemary oil should be classified as: STOT SE 2 (H371 / May cause damage to CNS following exposure) according to EU CLP (EC no 1272/2008 and its amendments).