Melaleuca viridiflora extract

Acute Toxicity: oral

Administrative data

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.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

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

Test material

Test material information

Constituent 1

Reference substance name:

Melaleuca viridiflora extract

EC Number:

310-217-5

EC Name:

Melaleuca viridiflora extract

Cas Number:

132940-73-9

IUPAC Name:

Essential oil of Melaleuca viridiflora (Myrtaceae) obtained from twigs and leaves by steam distillation

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): NIAOULI Essential Oil (Melaleuca quinquenervia) 021261
- Physical state: Colorless liquid
- Analytical purity: Not applicable (considered as 100 % for the study)
- Lot/batch No.: 1855343
- Production date: 23 January 2010
- Date of receipt: 06 September 2010
- Expiration date of the lot/batch: 23 January 2011
- Storage condition of test material: Room temperature

Test animals

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

Administration / exposure

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

Results and discussion

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.

Effect levels

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

Any other information on results incl. tables

None

Applicant's summary and conclusion

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).