Anisole

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 24 October 2001 and 05 December 2001.

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study. The purity of the test substance is not indicated.

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:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

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:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Anisole

EC Number:

202-876-1

EC Name:

Anisole

Cas Number:

100-66-3

Molecular formula:

C7H8O

IUPAC Name:

anisole

Test material form:

other: liquid stored at room temperature in the dark

Details on test material:

- Name of test material (as cited in study report): Anisole

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) Ltd, Margate, Kent
- Age at study initiation: eight to twelve weeks
- Weight at study initiation: 200 - 350 g
- Fasting period before study: no data
- Housing: in groups of five by sex.
- Diet (e.g. ad libitum): Rat and Mouse Expanded Diet No. 1, Special Diets Services Liited, Witham, Essex, UK OR or similar.
- Water (e.g. ad libitum): ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 2°C
- Humidity (%): 55 +/- 15%
- Air changes (per hr): at least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light

IN-LIFE DATES: no data

Administration / exposure

Route of administration:

inhalation: vapour

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: the animals were exposed to an atmosphere of the test item using a TSE Rodent Exposure System.
- Exposure chamber volume: 30 L
- Method of holding animals in test chamber: each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber. Only the nose of each animal was exposed to the test atmosphere.
- Method of conditioning air: no data
- System of generating particulates/aerosols: the particle size of the generated atmosphere inside the exposure chamber was determined three times during the exposure period using a Marple Personal Cascade Impactor (Schaefer Instruments Ltd, Oxon., UK). This device consisted of six impactor stages (9.8, 6.0, 3.5, 1.55, 0.93 and 0.52 µm cut points) with stainless steel collection substrates and a back up glass fibre filter, housed in an aluminium sampler. The collection substrates and backup filter were weighed before and after sampling and the weight of test material, collected at each stage, calculated by difference.
- Method of particle size determination: due to the high proportion of the chamber atmosphere estimated to be in vapour phase (95.6%), it was considered that no meaningful calculation of the atmosphere's characteristics (i.e. particle size, geometric standard deviation and respirable portion) was possible.
- Treatment of exhaust air: after passing through the animal’s breathing zone, spent aerosol entered the outer cylinder from where it was exhausted through a suitable filter system. Atmosphere generation was therefore dynamic.
- Temperature, humidity, pressure in air chamber: see Table 7.2.2/1

TEST ATMOSPHERE
- Brief description of analytical method used: the actual chamber concentration was measured at regular intervals during the exposure period. The sampling procedure involved pumping two litres of the chamber atmosphere through a glass impinger containing 40 mL of methanol. After sampling, the dreschel head was flushed through with a further 40 mL of methanol to remove any deposits. This gave an 80 mL sample to be submitted for chemical analysis. The nominal chamber concentration was calculated by dividing the mass of test material used by the total volume of air passed through the chamber.
- Samples taken from breathing zone: yes

VEHICLE: not applicable


TEST ATMOSPHERE (if not tabulated): not applicable


CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: limit test

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

- Nominal: 7.07 mg/L
- Mean Achieved: 6.51 mg/L (SD: 1.16 mg/L)

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: all animals were observed for clinical signs at hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, one hour after termination of exposure and subsequently once daily for fourteen days. Any evidence of avert toxicity was recorded at each observation. Individual bodyweights were recorded prior to treatment on the day of exposure and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: during necropsy, all animals were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded. The respiratory tract was subjected to a detailed macroscopic examination for signs of irritancy or local toxicity.

Statistics:

None

Results and discussion

Preliminary study:

Not applicable

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 6.51 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: No deaths occured during the study

Mortality:

No mortality was dectected during this study.

Clinical signs:

other: Signs of hunched posture, pilo-erection and red/brown staining around snout and/or eyes are commonly seen in animals for short periods on removal from the chamber following 4 hour inhalation studies. Wet fur is commonly recorded both during and for a shor

Body weight:

Normal bodyweights development was recorded during the study. Two females showed low bodyweight gain but such variations are not uncommon in female rats of this strain or age.

Gross pathology:

No macroscopic abnormalities were noted at necropsy.

Other findings:

None

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, Anisole did not induced acute toxicity after a single exposure of rat by inhalation (vapours, 4h). As the LC 50 in males and females was greater than 6.51 mg/L, the submitted substance is considered as not harmful by inhalation and is therefore not classified according to the Regulation (EC) 1272/2008 criteria.

Executive summary:

In an acute inhalation toxicity study, performed according to the Guideline 403 and in compliance with the GLP, groups of ten Sprague-Dawley Crl:CD (SD) IGS BR strain rats (5/sex) were exposed to Anisole for 4 hours at concentration of 7.07 mg/L (nominal) equivalent to the mean achieved atmosphere concentration of 6.51 mg/L (SD: 1.16 mg/l). The animals were exposed to an aerosol / vapour atmosphere using a nose only exposition. Animals then were observed for 14 days for mortality, clinical signs and bodyweight gain and then euthanized for gross necropsy.

No deaths occurred in this study. Respiratory rate, pilo-erection, hunched posture, and wet fur were observed and there were occasional instances of red/brown staining around the snout and/or eyes. There was no effect on bodyweight. No macroscopic abnormalities were noted at necropsy.

LC₅₀Male and Female rats/4 hours/ vapours: >6.51 mg/L.

Under the test conditions, Anisole did not induced acute toxicity after a single exposure of rat by inhalation (vapours, 4h). As the LC 50 in males and females was greater than 6.51 mg/L, the submitted substance is considered as not harmful by inhalation and is therefore not classified according to the Regulation (EC) 1272/2008 criteria.

This acute inhalation study is classified as acceptable even if higher concentrations than 6.51 mg/L were not tested. It satisfies the guideline requirement for an acute inhalation study in the rats.