Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: LD50>2000 mg/kg bw (similar to OECD 420)

Acute dermal toxicity: LD50=332 mg/kg bw (similar to OECD 402)

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:
25 Sept-16 Oct 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study is in accordance with GLP, according to OECD Guidelines No 420 "Acute Oral Toxicity - Fixed Dose Method" and (EC) 440/2008 No. B1 bis Acute Toxicity (Oral).
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)
Qualifier:
equivalent or similar to guideline
Guideline:
EU Method B.1 bis (Acute Oral Toxicity - Fixed Dose Procedure)
GLP compliance:
yes (incl. QA statement)
Test type:
fixed dose procedure
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: RccHan(TM):WIST (Harlan Laboratories, UK)
- Age at study initiation: 8-12 weeks, nulliparous and non-pregnant
- Weight at study initiation: 137-165 g
- Fasting period before study: overnight
- Housing:groups of four in suspended solid-floor polypropylene cages furnished with woodflakes
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum, 2014C Teklad Global Rodent diet (Harlan Laboratories, UK)
- Acclimation period: al least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25°C
- Humidity (%): 30-70%
- Air changes (per hr): al least 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hrs light/12 hrs dark

Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 1.91 ml/kg bw.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 females (of which 1 animal was tested in the sighting study)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: clinical observations were made 0.5, 1, 2 and 4 hours after dosing, and subsequently once a day for up to 14 days. Morbidity checks were made twice a day. Bodyweights were recorded on day 0, 7, 14 or at death.
- All surviving animals were killed humanely and subjected to gross necroscopy.
Preliminary study:
yes, one female at 2000 mg/kg bw
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 1/5 animals died during test
Mortality:
One animal was found dead one day after dosing in the main study. In the sighting study with one animal no mortality was observed.
Clinical signs:
Signs of systemic toxicity noted four hours after dosing in the initial treated animal were ataxia, hunched posture and pilo-erection. There were no signs of systemic toxicity noted in the remaining animals.
Body weight:
Surviving animals showed expected gains in bodyweight over the observed period.
Gross pathology:
Abnormalities noted at necropsy of the animal that died during the study were dark liver, dark kidney, gaseous stomach and pale gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bw.
Therefore, the substance does not need to be classified as acute toxic via the oral route based on the criteria as outlined in 1272/2008/EC (CLP).
Executive summary:

An acute oral toxicity study - fixed dose method - was performed with five female Wistar strain rats. Study was performed according to guidelines OECD No. 420 and EC 440/2008 No. B1 bis. Following a sighting test at a dose level of 2000 mg/kg bw, an additional four fasted animals were given a single dose of test item at a dose level of 2000 mg/kg bw. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy. One animals was found dead one day after dosing in the main study. Signs of systemic toxicity noted in the initial treated animal were ataxia, hunched posture and pilo-erection. There were no signs of toxicity in the remaining animals tested in the main study. Surviving animals showed expected gains in bodyweight over the observation period. Abnormalities noted at necropsy of the animal that died during the study were dark liver, dark kidney, gaseous stomach and pale gastric mucosa. No abnormalities were noted at necropsy of animals that were killed at the end of the study. The acute oral median lethal dose (LD50) of the test item in the female Wistar strain rat was estimated to be greater than 2000 mg/kg bw under the conditions of this test. Therefore, the substance does not need to be classified as acute toxic via the oral route based on the criteria as outlined in 1272/2008/EC (CLP).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

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

Additional information

Acute oral toxicity

The key study was a standard acute limit test, in which a single dose of Cassia oil (2000 mg/kg bw) was administered by oral gavage to 5 female Wistar rats. One animal was found dead one day after dosing, but no other mortality was observed up to 14 days after application. Based on these results, the oral LD50 was established to be >2000 mg/kg bw. A supporting study is available that confirms the LD50 is over 2000 mg/kg bw.

Acute dermal toxicity

Two studies were available for this endpoint, which were used in a Weight of Evidence approach:

The first available study was a standard acute test, in which 10 albino rabbits (2 per dose level) were exposed to a single dose of Cassia oil (168, 332, 600, 1323, 2625 mg/kg bw, calculated from mL/kg based on density). At the lowest dose none of the animals died, but at the dose level of 332 mg/kg bw, 1 of two animals (50%) died. At the higher dose levels all animals died. Based on the mortality rates the dermal LD50 was established to be 332 mg/kg bw.

The second study was performed according to an equivalent of guideline OECD 402 (acute dermal toxicity) but with a deviating dose (1000 mg/kg bw instead of 2000 mg/kg bw). 5 male and 5 female New Zealand White rabbits were selected. A single dose of the test article was applied for 24 hours. Eight of ten animals survived. Two males died on day 3 of the observation period. The LD50 was therefore determined to be greater than 1000 mg/kg body weight.

Based on the results of these two studies (WoE approach) it was decided to use the lowest reported LD50 of 332 mg/kg bw as the starting point for classification.

Justification for selection of acute toxicity – oral endpoint

The selected study is the key study for this endpoint.

Justification for selection of acute toxicity – dermal endpoint

No selection is made as a Weight of Evidence approach was followed which is described below.

Justification for classification or non-classification

Based on the available information, Cassia oil has shown to be non-toxic after oral exposure. Therefore, the substance does not need to be classified for Acute Oral Toxicity in accordance with the criteria outlined in Annex I of 1272/2008/EC (CLP/EU-GHS).

Based on the available information, Cassia oil has shown to be toxic in contact with skin. Therefore, the substance needs to be classified for Acute Dermal Toxicity (Category 3 / H311) in accordance with the criteria outlined in Annex I of 1272/2008/EC (CLP/EU-GHS).