Myristica fragrans, ext.

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

6 December 1984 - 19 December 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Test was conducted according to EPA proposed guidelines for registering pesticides in the U.S.; Hazard Evaluation: Humans and Domestic Animals: Federal Register (43) 163, August 22, 1978. 163 81.3, and equivalent to OECD Test Guideline 403,1981, under GLP Standards, and QA. Chemical identity of test substance is not reported.

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:

1985

Report date:

1985

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPP 81-3 (Acute inhalation toxicity)

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Myristica fragrans, ext.

EC Number:

282-013-3

EC Name:

Myristica fragrans, ext.

Cas Number:

84082-68-8

Molecular formula:

Unspecified (UVCB)

IUPAC Name:

Nutmeg Oil (Myristica fragrans, ext.)

Constituent 2

Reference substance name:

84-215-01

IUPAC Name:

84-215-01

Details on test material:

- Name of test material (as cited in study report): 84-215-01
- Physical state: liquid
- Analytical purity: no data
- Lot/batch No.: confidential
- Storage condition of test material: at room temperature

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited, Manston Road, Margate, Kent, UK
- Age at study initiation: no data
- Weight at study initiation: 190 to 220 (mean)
- Fasting period before study: no data
- Housing: placed in an individual wire mesh holding cage. The cages were made of stainless steel mesh (size 30.5 cm x 19 cm x 20 cm height) and were suspended on a movable rack. The rats remained in a holding room except for the 4-hour exposure.
- Diet: food (Scientific feeds LAD 1) ad libitum
- Water: tap water ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Max. 24°C (SD 0.7) and min. 23°C (SD 0.9)
- Humidity (%): 46% (SD 7.7)

Administration / exposure

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

air

Remarks:

15 l/min.

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Exposure chamber made of perspex
- Exposure chamber volume: 0.13m3
- Method of holding animals in test chamber: The whole body exposure chambers used for the exposures were of square section and were fitted with pyramidal tops. Each chamber was divided by wire mesh partitions to provide 10 separate animal compartments. The rats (5 male and 5 female) were placed into separate compartments of the exposure chamber.
- Source and rate of air: concentric jet atomiser - clean air: 10 l/min.; diluent air: 15 l/min.
vapour generator - test substance: 0.26 ml/min.
- System of generating particulates/aerosols: The test vapour was generated using a Pyrex glass with a condenser through which warm water was passed and the delivery system of the test substance (injected into a glass concentric jet atomizer with a syringe pump). As the spray of the test substance passed through the condenser (kept at 48 °C), some of it evaporated and was passed through a glass tube and diluted to the appropriate concentration of 5.0 mg/l.
- Temperature, humidity, pressure in air chamber: The temperature in the exposure chamber was measured with a mercury bulb thermometer and recorded at the start of the exposure and at 30-minute intervals during the exposure. The chamber relative humidity was monitored continuously during exposure using a wet and dry bulb hygrometer and recorded at 30-minute intervals.Temperature (°C): Control group: 23.8 (0.75), Test group: 25.1 (0.46); Relative humidity: Control group: 34% (5.5%), Test group:45% (1.7%).

TEST ATMOSPHERE
- Brief description of analytical method used: Ten air samples were taken from the chamber during each exposure and analysed to determine the concentration of vapour from the test substance in the chamber atmosphere.The test atmosphere was drawn through a glass tube containing OV17 5% + 1% Dexil in order to trap the test vapour which was then thermally desorbed into a gas chromatograph equipped with flame ionisation detector. The areas of the peaks due to the test vapour were then used to calculate the atmospheric concentrations.
- Samples taken from breathing zone: no

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

ca. 4 h

Concentrations:

The mean concentration of vapour from the test substance to which the rats were exposed was 4.85 mg/1 and the variation ((range x 100) / mean) was 23%. Measured concentrations are included in the field "any other information on materials and methods incl. tables".

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed continuously during exposure, at hourly intervals for 4 hours following exposure and at least twice daily during the observation period. In addition to the range of clinical signs normally recorded, behavioural parameters were scored or measured once daily for 3 days before exposure and daily throughout the observation period. Each animal was weighed on the day of dosing (Day 1) and on Days 2, 4, 7, 11 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical and behavioral signs, body weight, rectal temperatures, organ weights (liver, heart, kidneys, adrenals, and lungs), gross necropsy

Statistics:

Analyses of variance were carried out on the following data: bodyweights, rectal temperatures, organ weights. Students' 't' distribution was used to compare the treatment means.

Results and discussion

Preliminary study:

Not relevant

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4.9 mg/L air (analytical)

Exp. duration:

4 h

Mortality:

No deaths were seen in any animal of the test article or air only control groups.

Clinical signs:

other: The clinical signs seen during exposure were consistent with the response to a mildly irritant aerosol and included salivation, lacrimation and adoption of an abnormal respiratory pattern and body posture. Peripheral vasodilation was also seen in 2/5 male

Body weight:

The rate of bodyweight gain for control and test groups was similar.

Gross pathology:

All rats were normal with the exception of one treated male rat. This rat had a few petechiae on the lung surface.

Other findings:

- Organ weights: No significant test article related effects are noted.
- Other observations: the following changes were considered spontaneous in origin and of no toxicological significance: Bronchiolitis/bronchiolar epithelial erosion/bronchiolar epithelial hyperplasia in 2 control rats (male, female) and 1 treated female rat. Focus of pneumonitis in 1 control female rat and 1 treated male rat.

Any other information on results incl. tables

Not relevant

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In this acute inhalation toxicity study the 4-hour LC50 of Nutmeg oil in the rat is determined to be greater than 4.9 mg/1. Based on these results, the test substance does not need to be classified for acute inhalation toxicity according to the EU criteria outlined in Annex I of 1272/2008/EC and Annex VI of 67/548/EEC.

Executive summary:

This acute inhalation toxicity study in male and female rats was conducted according to a method equivalent to OECD test guideline 403. Two groups of 10 rats (5/sex/group) were exposed whole-body to either 4.9 mg/1 vapour from test article 84-215-01 (nutmeg oil) or air only for 4 hours as control. Ten air samples are taken and test article concentration is determined by gas chromatography. Animals are observed once per hour for the first 4 hours during exposure and at least twice daily for 14 days, thereafter. No deaths were seen in any animal of the test article or air only control groups. Test article related clinical signs to test article exposure are irritant effects, abnormal respiratory pattern and abnormal body position. A behavioural evaluation yields no significant effects for either group. Slightly lower body temperatures follow the exposure. No significant test article related effects are noted for the following parameters: body weight, organ weights, microscopic and macroscopic pathology. The 4-hour LC50 of Nutmeg oil in the rat is determined to be greater than 4.9 mg/1. Based on these results, the test substance does not need to be classified for acute inhalation toxicity according to the EU criteria outlined in Annex I of 1272/2008/EC and Annex VI of 67/548/EEC.