Melaleuca viridiflora extract

Endpoint summary

Description of key information

Acute toxicity, oral: LD50 cut-off = 2500 mg/kg bw (K).

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:

14 September – 05 October 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well conducted and well described study in accordance with GLP and OECD Guideline 423 without any deviation.

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

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Elevage Janvier, Le Genest-St Isle, France
- Age at study initiation: 8 weeks
- Weight at study initiation: 182-210 g
- Fasting period before study: Food was removed on Day 1 and then redistributed 4 h after administration of the test item.
- Housing: Animals were housed by group of three in solid bottomed clear polycarbonate cages with stainless steel mesh lid.
- Diet: Foodstuff (M20-SDS), ad libitum
- Water: Drinking water (tap water from public distribution system), ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 19-25 °C
- Humidity: 30-70 %
- Air changes: Approximately 15 changes/h
- Photoperiod: 12 h dark / 12 h light

Route of administration:

oral: gavage

Vehicle:

other: test item was used as supplied in first step; 2nd and 3rd step: 2.21 mL of test item was added to 3.33 mL of olive oil

Details on oral exposure:

MAXIMUM DOSE VOLUME APPLIED: 5.54 mL/kg bw

DOSAGE PREPARATION:
- In the first step of the study, the test item was administered as supplied at 5.54 mL/kg bw (corresponding to 5000 mg/kg bw according to calculated density). In the second and third step of the study, 2.21 mL of the test item (corresponding to 2000 mg/kg bw according to calculated density) was added to 3.33 mL of olive oil to obtain a yellow solution just before administration.

Doses:

- Step 1: 5000 mg/kg bw
- Step 2: 2000 mg/kg bw
- Step 3: 2000 mg/kg bw

No. of animals per sex per dose:

- Step 1: 1 female (5000 mg/kg bw)
- Step 2: 3 females (2000 mg/kg bw)
- Step 3: 3 females (2000 mg/kg bw)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Clinical signs and mortality: Animals were examined every day for 14 days.
Bodyweight: Animals were weighed on Days 0 (just before administering the test item), 2, 7 and 14.
- Necropsy of survivors performed: Yes; On Day 14, the animals were anesthetized with sodium pentobarbital and administration continued to fatal levels and macroscopic examination was performed.

Statistics:

None

Preliminary study:

At 5000 mg/kg bw, animal was died (1/1) at 7 hours 5 minutes post-dose. The mortality was preceded by an increase in salivation, lachrymation, absence or decrease in spontaneous activity, Preyer's reflex, body temperature, muscle tone and righting reflex, bradypnea, and staggering gait after handling or animal lying on the floor. The dead animal showed a rigor mortis during macroscopical examination, which revealed thinning of the corpus, white coloration of the forestomach and yellowish liquid in the intestines.

Sex:

female

Dose descriptor:

LD50

Effect level:

2 500 mg/kg bw

Based on:

test mat.

Remarks on result:

other: LD50 cut-off according to OECD Test Guideline No. 423 testing scheme (one animal was died (1/6) at 2000 mg/kg bw)

Mortality:

- One animal was died (1/6) at 2000 mg/kg bw on Day 3.
- Mortality was preceded by an absence or a decrease in spontaneous activity, Preyer's reflex, body temperature, muscle tone and righting reflex, bradypnea, partial ptosis, myosis, increased lachrymation, and a staggering gait after handling or animal lying on the floor. The mortality was also preceded by decrease in the body weight of the animal: -9 % on Day 2 compared to Day 0.

Clinical signs:

other: In the survival animals treated at 2000 mg/kg bw, decrease in spontaneous activity (4/5), increased lachrymation (3/5), staggering gait after handling (4/5), and piloerection (3/5) were noted on the first day of the study. The animals recovered a normal b

Gross pathology:

- At 2000 mg/kg bw, macroscopical examination of the animals at the end of the study did not reveal treatment-related changes.
- The dead animal showed a rigor mortis at the time of the macroscopical examination, which revealed a brown coloration of the corpus, a thinning of the forestomach, and yellow coloration of the intestines.

Other findings:

None

None

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the test conditions and according to the OECD Test Guideline No. 423 testing scheme,, the oral LD50 cut-off for NIAOULI Essential Oil (Melaleuca quinquenervia) 021261 is 2500 mg/kg bw in female rats therefore it is not classified according to the Annex VI to the Directive 67/548/EEC and the Regulation (EC) No 1272/2008 (CLP).

Executive summary:

In an acute oral toxicity study performed according to OECD Guideline 423 and in compliance with GLP, 6 female Sprague Dawley rats were given a single oral (gavage) dose of NIAOULI Essential Oil (Melaleuca quinquenervia) 021261 at 2000 mg/kg bw. Animals were then observed for mortality, clinical signs and bodyweights for 14 days and were all sacrificed for macroscopic examination. Preliminary study was conducted with one female rat at 5000 mg/kg bw.

In preliminary study, one animal was died (1/1) at 7 hours 5 minutes post-dose. In main study, one animal was died (1/6) at 2000 mg/kg bw on Day 3. Mortality was preceded by an absence or a decrease in spontaneous activity, Preyer's reflex, body temperature, muscle tone and righting reflex, bradypnea, partial ptosis, myosis, an increased lachrymation, and a staggering gait after handling or animal lying on the floor. The mortality was also preceded by decrease in the body weight of the animal: -9 % on Day 2 compared to Day 0.

In the survival animals treated at 2000 mg/kg bw, decrease in spontaneous activity (4/5), increased lachrymation (3/5), staggering gait after handling (4/5), and piloerection (3/5) were noted on the first day of the study. The animals recovered a normal behaviour at 48 h post-dose.

One animal was lying on the floor at 24 hours, examinations revealed an absence of Preyer's reflex, decreased body temperature and muscle tone, absence of righting reflex, bradypnea, partial ptosis and myosis. The animal recovered a normal behaviour at 48 h post-dose.

A slight decrease in the body weight gain was noted in the animals treated at 2000 mg/kg bw on Day 2: +5 % on Day 2 compared to Day 0, versus +11 % in the historical control group. Macroscopical examination of the animals at the end of the study did not reveal treatment-related changes. The dead animal showed a rigor mortis at the time of the macroscopical examination, which revealed a brown coloration of the corpus, a thinning of the forestomach, and yellow coloration of the intestines.

 

Under the test conditions and according to the OECD Test Guideline No. 423 testing scheme, the oral LD50 cut-off for NIAOULI Essential Oil (Melaleuca quinquenervia) 021261 is 2500 mg/kg bw in female rats therefore it is not classified according to the Annex VI to the Directive 67/548/EEC and the Regulation (EC) No 1272/2008 (CLP).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 500 mg/kg bw

Quality of whole database:

The key study is GLP-compliant and of high quality (Klimisch score = 1)

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute toxicity: via oral route A key study was identified (Colas, 2010, Rel.1). In this acute oral toxicity study performed according to OECD Guideline 423 and in compliance with GLP, rats were given a single oral (gavage) dose of NIAOULI Essential Oil at 2000 mg/kg bw. In the preliminary study, the animal dosed with 5000 mg/kg bw died within 7 hours. In the main study, 1/6 animal dosed with 2000 mg/kg bw was found dead on Day 3. Mortality was preceded by an absence or a decrease in spontaneous activity, Preyer's reflex, body temperature, muscle tone and righting reflex, bradypnea, partial ptosis, myosis, an increased lachrymation, and a staggering gait after handling or animal lying on the floor. The mortality was also preceded by decrease in the body weight of the animal: -9 % on Day 2 compared to Day 0. In the survival animals treated at 2000 mg/kg bw, decrease in spontaneous activity (4/5), increased lachrymation (3/5), staggering gait after handling (4/5), and piloerection (3/5) were noted on the first day of the study. The animals recovered a normal behaviour at 48 h post-dose. One animal was lying on the floor at 24 hours, examinations revealed an absence of Preyer's reflex, decreased body temperature and muscle tone, absence of righting reflex, bradypnea, partial ptosis and myosis. The animal recovered a normal behaviour at 48 h post-dose. A slight decrease in the body weight gain was noted in the animals treated at 2000 mg/kg bw on Day 2: +5 % on Day 2 compared to Day 0, versus +11 % in the historical control group. Macroscopical examination of the animals at the end of the study did not reveal treatment-related changes. The dead animal showed a rigor mortis at the time of the macroscopical examination, which revealed a brown coloration of the corpus, a thinning of the forestomach, and yellow coloration of the intestines. In this study and according to the OECD Test Guideline No. 423 testing scheme, the oral LD50 cut-off of test item was considered to be 2500 mg/kg bw in female rats.

Justification for selection of acute toxicity – oral endpoint
Only one study available

Justification for classification or non-classification

Harmonized classification:

Niouli oil has no harmonized classification according to the Regulation (EC) No. 1272/2008.

Self-classification:

Acute toxicity via Oral route:

Based on the available information, Niaouli oil is:

- not classified according to the Regulation (EC) No. 1272/2008 as the LD50 is greater than 2000 mg/kg bw

- not classified according to the Directive 67/548/EEC as the LD50 is greater than 2000 mg/kg bw.

Acute toxicity via Dermal route: This information is not available

Acute toxicity via Inhalation: This information is not available.

Specific target organ toxicity: single exposure (Oral):

The classification criteria according to the Annex VI of the Regulation (EC) No. 1272/2008 as specific target organ toxicant (STOT) – single exposure, oral are not met since no reversible or irreversible adverse health effects were observed immediately or delayed after exposure and no effects were observed at the guidance value (oral) for a Category 1 classification (C ≤ 300 mg/kg bw) and at the guidance value (oral) for a Category 2 classification (2000 mg/kg bw ≥ C > 300 mg/kg bw). No classification is required.

Specific target organ toxicity: single exposure (Dermal): This information is not available

Specific target organ toxicity: single exposure (Inhalation): This information is not available.

Based on its composition (> 10% of aspiration toxicants, i.e. limonene, pinene), Niaouli oil should be classified for aspiration hazard:

- H304,May be fatal if swallowed and enters airways according to the regulation (EC) No. 1272/2008

- Xn, R65, Harmful: may cause lung damage if swallowed according to the Directive 67/548/EEC.

Source: ECHA disseminated dossiers

-Pinene:

http://apps.echa.europa.eu/registered/data/dossiers/DISS-9d952924-c8ed-4614-e044-00144f67d249/DISS-9d952924-c8ed-4614-e044-00144f67d249_DISS-9d952924-c8ed-4614-e044-00144f67d249.html

- Limonene:

http://apps.echa.europa.eu/registered/data/dossiers/DISS-9eb16d5d-b83e-2831-e044-00144f67d031/DISS-9eb16d5d-b83e-2831-e044-00144f67d031_DISS-9eb16d5d-b83e-2831-e044-00144f67d031.html