Anisole

Endpoint summary

Description of key information

Anisole is not harmful by any of the three routes of exposure: 
- Oral LD50 in male and female rats was 3700 mg/kg bw (95% confidence limits: 3240-4220 mg/kg bw).
- Oral LD50 of Anisole to male white mice by gavage was 2800 mg/kg bw.
- Dermal LD50/rabbits > 5000 mg/kg bw
- Inhalation LC50 Male and Female rats/4 hours/ vapours: >6.51 mg/L. 

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

No data

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Study performed on 1946, no details were available but conducted according to accepted scientific methods.

Qualifier:

no guideline available

Principles of method if other than guideline:

No data

GLP compliance:

no

Remarks:

(before GLP statement)

Test type:

standard acute method

Limit test:

no

Species:

mouse

Strain:

not specified

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation:20-30 grams.
- Housing: Mice were housed individually in wire cages.
- Diet (e.g. ad libitum): Fed Purina dog chow with each dose.

Route of administration:

oral: gavage

Vehicle:

olive oil

Details on oral exposure:

The calculated doses were introduced into the stomach through the esophagus by means of a tuberculin syringe with a blunt needle.

Doses:

5, 10, and 25% solutions in olive oil.

No. of animals per sex per dose:

10 males per dose

Control animals:

not specified

Details on study design:

- Duration of observation period following administration: 1 week
- Other examinations performed: Mice were observed for tremors, convulsions, and fatalities.

Statistics:

No data

Preliminary study:

No data

Sex:

male

Dose descriptor:

LD50

Effect level:

2 800 mg/kg bw

Based on:

test mat.

Mortality:

four of eight animals died at a dose of 2800 mg/kg.

Clinical signs:

other: Only moderate depression was caused with lower doses while higher doses primary excitement then progressive depression, rapid and labored respiration and finally coma and death.

Gross pathology:

No data

Other findings:

Animals showed marked cyanosis in the final stages of poisoning.

None

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

The oral LD50 of Anisole to male white mice by gavage was determined to be 2800 mg/kg bw. Based on this study, Anisole is not classified according to EU GHS criteria and is classified Acute. Tox. Cat. 5, H303 according to UN GHS criteria.

Executive summary:

The acute oral toxicity of Anisole was determined in mice. The test was administered to 10 males per dose by gavage route at 5, 10, and 25% solutions in olive oil. The animals were observed over a 1-week period for tremors, convulsions, and fatalities. No further details were provided.

Four of eight animals died at a dose of 2800 mg/kg. Only moderate depression was caused with lower doses while higher doses primary excitement then progressive depression, rapid and labored respiration and finally coma and death.

The oral LD50 of Anisole to male white mice by gavage was determined to be 2800 mg/kg bw.

Based on these results, Anisole is not classified for acute toxicity according to the Regulation (EC) 1272/2008 (CLP).

 

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

No data

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Few information are available about the test conditions and the results. No information about the test susbtance purity.

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline available

Principles of method if other than guideline:

Groups of Osborne-Mendel male and female rats (5 animals/sex/dose) were given a single oral dose of Anisole by gavage route. Clinical signs, mortality and/or systemic effects were checked for a period of up to 14 days. LD50 results were calculated per Litchfield-Wilcoxon (1949). No further details were provided.

GLP compliance:

no

Remarks:

study performed before GLP statement.

Test type:

other: no data

Limit test:

no

Species:

rat

Strain:

Osborne-Mendel

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: no data
- Age at study initiation: not indicated (young adult)
- Weight at study initiation: 180-350 g
- Fasting period before study: 18 hours
- Housing: no data
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

No data

Doses:

No data

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: no data
- Necropsy of survivors performed: no data
- Other examinations performed: clinical signs, body weight and/or systemic effects

Statistics:

The Litchfield and Wilcoxon method (1949) was used to calculate the LD50.

Preliminary study:

Not applicable

Sex:

male/female

Dose descriptor:

LD50

Effect level:

3 700 mg/kg bw

Based on:

test mat.

95% CL:

3 240 - 4 220

Remarks on result:

other: Slope = 1.2 (1.1-1.4)

Mortality:

Death time between 4 hrs and 8 days.

Clinical signs:

other: Depression, porphyrin like deposit on eyes, salivation, blood urine and rough fur.

Gross pathology:

no data

Other findings:

no data

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Under the test conditions, Anisole is not classified according to EU GSH criteria for acute oral toxicity as LD50 male/female rats was 3700 mg/kg bw.
According to UN GHS criteria, Anisole is classified: Acute. Tox. cat. 5, H303.

Executive summary:

The acute oral toxicity of Anisole was determined in rats of the Osborne-Mendel strain. The test substance (purity unknown) was administered to 5 male and 5 female rats per dose by gavage route. The fasting period before study was 18 hours. The animals were observed over a 2-week period for mortality and/or systemic effects. LD50 results were calculated per Litchfield-Wilcoxon (1949). No further details were provided.

Oral LD50 in male and female rats was 3700 mg/kg bw (95% confidence limits: 3240-4220 mg/kg bw).

Deaths occurred from 4 hours - 8 days. Depression, porphyrin like deposit on eyes, salivation, blood urine and rough fur were observed.

Based on these results, Anisole is not classified for acute toxicity according to the Regulation (EC) 1272/2008 (CLP).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

Two acute oral studies in rats and mice are available. However, few details are described (Reliability 4). A waiving was proposed.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

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.

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 403 (Acute Inhalation Toxicity)

Deviations:

no

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

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

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

Preliminary study:

Not applicable

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

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

Key study according to OECD 403 with reliability 2.The OECD 412 range finding is a sub-acute study (7 days).

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Review of the different end-points of Anisole toxicity (secondary source). The initial reference is not published.

Interpretation of results:

GHS criteria not met

Executive summary:

A dermal LD50 study was conducted in which 5000 mg/kg of Anisole was applied to the skin of 10 rabbits. The animals were observed for mortality and/or systemic effects for 14 days. No further details were provided.

Acute dermal LD50/rabbits > 5000 mg/kg bw was determined.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

Only one study was available. However, few details are described (Reliability 4). A waiving was proposed.

Additional information

Acute oral toxicity:

Two acute oral studies in rats and mice are available. However, only few details are described (reliability 4):

- Oral LD50 in male and female rats was 3700 mg/kg bw (95% confidence limits: 3240-4220 mg/kg bw).

- Oral LD50 of Anisole to male white mice by gavage was determined to be 2800 mg/kg bw.

Based on the results of these studies, Anisole was considered as not harmful via oral route.

Taking into account these results and that the exposure of humans is only by inhalation route, another acute test by oral route is not necessary.

Acute dermal toxicity:

Only one study was available in rabbit with reliability 4. In this study, acute dermal LD50/rabbits > 5000 mg/kg bw was determined. Based on this result, Anisole was considered as not harmful via dermal route.

Taking into account this result and that the exposure of humans is only by inhalation route, another acute test by dermal route is not necessary.

Acute inhalation toxicity:

Only one acute toxicity study by inhalation was available (OECD 403, reliability 2) and was considered as the key study.

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. 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 was determined.

The range finding study of sub-acute inhalation study (OECD 412, reliability 1) was considered as a supporting study. Groups of 3 male and 3 females were exposed to anisole at 0 (control), 1, 3 or 10 g/m3. Exposure to concentrations up to 3 g/m3 did not result in any significant treatment-related changes. Exposure to 10 g/m3, which was reduced to 6 g/m3 after the first exposure in response to the animals’ poor condition, resulted in many clinical abnormalities,such as ataxia, lethargy, dyspnoea and piloerection, decreased body weight gain and food consumption, increased relative (to body weight) liver weight in males and females, and increased weight of the adrenals in female animals. But no mortality occured and detailed clinical observations prior to necropsy did not reveal any exposure-related neurobehavioural toxicity.

In conclusion, based on the results of these studies, Anisole is considered as not harmful via inhalation route.

Justification for classification or non-classification

Acute oral toxicity:

As the LD50/rat is 3700 mg/kg bw and the LD50/mice is 2800 mg/kg bw, Anisole is considered as not harmful by oral route. Therefore, no classification is required according to the Regulation (EC) 1272/2008 criteria.

Acute Dermal toxicity:

Since Acute dermal LD50/rabbits > 5000 mg/kg bw, Anisole is considered as not harmful by dermal route. Therefore, no classification is required according to the Regulation (EC) 1272/2008 criteria.

Acute Inhalation toxicity:

- The LC50 calculated in male and female rats is higher than 6.51 mg/L. Based on this value and according to the Regulation (EC) 1272/2008 (CLP) and the Directive 67/548/EEC criteria, Anisole is not considered as harmful if inhaled. This conclusion was confirmed by the results described in the range finding of the sub-acute inhalation toxicity study (OECD 412).

- In rat, exposure to 10 or 6 g/m3 of Anisole, resulted in many clinical abnormalities, such as ataxia and lethargy. These abnormalities were observed shortly after exposure and were transient. These effects were observed only in the range finding study (7 days) and not in the main study (OECD 412 - 29 days). Therefore, based on the 7 day study Anisole may cause drowsiness or dizziness and classified into category 3, H336 according to the Regulation (EC) 1272/2008 criteria.