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Description of key information

Oral: LD50 between 2060 and 5015 mg/kg bw, male/female mice, Smith (1979).
Dermal: LD50 > 2000 mg/kg bw, male/female rat, OECD 402, EU Method B.3, Sanders (2012).

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:
09 July 1979 to 07 September 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: A non-GLP study performed in accordance with generally accepted scientific principles with incomplete reporting and methodological deficiencies which do not affect the quality of the relevant results.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Four to five week old white mice were dosed with 3 dose levels using graded volumes of the test material via gavage. Animals were observed for up to 7 days after exposure. All animals dying were autopsied and those surviving until termination were sacrificed and examined post mortem after 7 days.
GLP compliance:
no
Test type:
other: Oral intubation at 3 dose levels
Limit test:
no
Species:
mouse
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 4-5 weeks.
- Fasting period before study: 4 hours.
- Housing: animals were placed in individual cages.
- Diet: commercial pelleted diet ad libitum.
- Water: ad libitum.
Route of administration:
oral: gavage
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
The animals were placed in individual cages, fasted for four hours and then the animals were individually weighed and inbtubated with the appropriate volumes of the test material. Each animal on one dose level received the same amount per kg body weight.

Doses:
2, 5 and 10 mL/kg bw.
No. of animals per sex per dose:
One male and one female were dosed with 2 and 10 mL/kg bw of the test material.
Three males and three females were dosed with 5 mL/kg bw of the test material.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 7 days.
- Frequency of weighing: Animals were weighed prior to dosing, and then again prior to sacrifice.
- Necropsy of survivors performed: yes.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 - < 5 mL/kg bw
Based on:
act. ingr.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 060 - 5 150 mg/kg bw
Based on:
act. ingr.
Mortality:
Mortalities can be seen in Table 1.
Clinical signs:
Mice dosed at all three dose levels were drowsy, showing signs of stress, and exhibiting laboured breathing within 30 minutes. Within 1 hour after treatment mice were also hypothermic. Within 2 hours of treatment the mice dosed at 10 mL/kg were comatose and those animals dosed at 2 and 5 mL/kg were semi-comatose. The surviving female mice recovered within 48 hours following treatment.
Body weight:
The surviving female mice gained weight during the 7 day observation period.
Gross pathology:
Autopsy of the animals that died revealed distension of the stomach due to the presence of orange/yellow fluid. The stomach and intestines were very pale and the ileum was irritated and haemorrhaging present in some animals. These mice also exhibited pale livers and kidneys. The surviving females appeared normal at autopsy.

Table 1. Mortalities

Dose Level ml/kg bw

No. of Dead Mice/No. of Animals Treated

10.0

2/2

5.0

4/6

2.0

1/2

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the test the approximate LD50 of the test material was determined to be between 2.0 and 5.0 mL/kg bodyweight.
Executive summary:

The acute oral toxicity of the test material was determined by gavage in 4-5 week old white mice at 3 dose levels using graded volumes of the test material, 10.0, 5.0 and 2.0 mL/kg bw. Animals were observed for up to 7 days after exposure. All animals dying were autopsied and those surviving until termination were sacrifices and examined post mortem after 7 days. Two mice were dosed at the high and low dose levels and six for the middle dose level. Equal numbers of males and females were used.

During the study both mice dosed with 10 mL/kg, four of the six mice dosed with 5 mL/kg and the one male mouse dosed at 2 mL/kg died within 18-24 hours of treatment. Mice dosed at all three dose levels were drowsy, showing signs of stress, and exhibiting laboured breathing within 30 minutes. Within 1 hour of treatment mice were hypothermic. Within 2 hours of treatment the mice dosed at 10 mL/kg were comatose and those animals dosed at 2 and 5 mL/kg were semi-comatose. The surviving female mice recovered within 48 hours following treatment. Autopsy of the animals that died revealed distension of the stomach due to the presence of orange/yellow fluid. The stomach and intestines were very pale and the ileum was irritated and haemorrhaging present in some animals. These mice also exhibited pale livers and kidneys. The surviving females were normal in appearance at autopsy.

The dose level was converted from mL/kg bw to mg/kg bw, for classification, based on the specific gravity of the test material reported in this study.

Under the conditions of the study the approximate LD50 of the test material was determined to be between 2060 and 5015 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The key study was determined to be reliable with restrictions. The data taken from the supporting studies are in good agreement with the results of the key study and are sufficient to support the determined LD50 value. Collectively representing a dataset with good quality overall.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 June 2012 to 11 July 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to valid guidelines and the study was conducted under GLP conditions.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 8 - 12 weeks.
- Weight at study initiation: At least 200 g.
- Housing: Individually during exposure period and in groups of up to four, by sex, for the remainder of the study.
- Diet: Rodent diet ad libitum.
- Water: Mains drinking water, ad libitum.
- Acclimation period: at least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 25 ºC
- Humidity (%): 30 - 70 %
- Air changes (per hr): at least 15 changes per hour.
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light.
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Backs and flanks of each animal, prepared by clipping free of hair the day before exposure.
- % coverage: Approximately 10% of the total body surface area.
- Type of wrap if used: A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage.

REMOVAL OF TEST SUBSTANCE
- Washing: After the 24 hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with arachis oil BP to remove any residual test material.
- Time after start of exposure: 24 hours after exposure.

TEST MATERIAL
- Amount applied: 1.93 mL/kg

TEST PROCEDURE
- Initially one male and one female were tested to assess the toxicity of the test material. Animals were dosed in the same manner as the other test animals. No mortalities were observed so the remaining animals were dosed. The results were combined for evaluation.
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
5 males and 5 females.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed for deaths or overt signs of toxicity 0.5, 1, 2 and 4 hours after dosing and once daily thereafter for 14 days. Individual bodyweights were recorded prior to application of the test material on day 0 and on days 7 and 14.
- Necropsy of survivors performed: Yes (external examination and opening the abdomen and thoracic cavities to check for any macroscopic abnormalities).
- Other examinations performed: Following removal of the test material the test sites were examined for evidence of primary skin irritation and scored according to Draize (1977) (See "Any other information on materials and methods"), any other skin reactions, if present were also recorded.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: There were no deaths and no signs of overt toxicity.
Mortality:
There were no deaths.
Clinical signs:
There were no signs of systemic toxicity.
Body weight:
All animals showed expected gains in bodyweight over the study period. Individual body weights can be seen in Table 2.
Gross pathology:
A cavity was noted in the right kidney of one male but this was considered to be a genetic defect and not related to treatment with the test material. No abnormalities were noted at necropsy of the remaining animals.
Other findings:
Dermal irritation: There were no signs of dermal irritation.

Table 2: Individual Bodyweights

Dose Level mg/kg Animal Number and Sex Bodyweight (g) at Day Bodyweight Change (g) During Week)
0 7 14 1 2
2000 1-0 Male 275 294 316 19 22
3-0 Male * 242 272 312 30 40
3-1 Male 236 272 310 36 38
3-2 Male 247 277 308 30 31
3-3 Male 242 277 313 35 36
2-0 Female 206 210 215 4 5
4-0 Female 215 225 232 10 7
4-1 Female 218 220 228 2 8
4-2 Female 218 227 230 9 3
4-3 Female 208 219 226 11 7

*Individual with the cavity in the right kidney.

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the study, the acute dermal LD50 was determined to be greater than 2000 mg/kg bw.
Executive summary:

The acute dermal toxicity of the test material was determined in a study conducted in compliance with standardised guidelines OECD 402 and EU Method B.3. During the study, two animals (one male and one female) were given a single, 24 -hour semi-occluded dermal application of the undiluted test material to intact skin at a dose level of 2000 mg/kg bw. Based on the results of the initial test, a further group of four males and four females was similarly treated. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Under the conditions of the study, there were no deaths or overt signs of systemic toxicity. There were no signs of dermal irritation and all animals showed expected gains in bodyweight over the study period. At necropsy a cavity was noted in the right kidney of one male but this was considered to be a genetic defect and not related to treatment with the test material. No abnormalities were noted at necropsy of the remaining animals. In consideration of these findings the LD50 was determined to be greater than 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The key study was determined to be reliable without restriction. Further supporting information was in good agreement with results from the key study, indicating a good overall quality of the database.

Additional information

Oral

In the key study (Smith, 1979) the acute oral toxicity of the test material was determined by gavage in 4-5 week old mice, at 3 dose levels; 10.0, 5.0 and 2.0 mL/kg bw. Animals were observed for up to 7 days after exposure. All animals dying were autopsied and those surviving until termination were sacrifices and examined post mortem after 7 days. Two mice were dosed at the high and low dose levels and six for the middle dose level. Equal numbers of males and females were used. During the study both mice dosed with 10 mL/kg, four of the six mice dosed with 5 mL/kg and the one male mouse dosed at 2 mL/kg died within 18-24 hours of treatment. Mice dosed at all three dose levels were drowsy, showing signs of stress, and exhibiting laboured breathing within 30 minutes. Within 1 hour of treatment mice were hypothermic. Within 2 hours of treatment the mice dosed at 10 mL/kg were comatose and those animals dosed at 2 and 5 mL/kg were semi-comatose. The surviving female mice recovered within 48 hours following treatment. Autopsy of the animals that died revealed distension of the stomach due to the presence of orange/yellow fluid. The stomach and intestines were very pale and the ileum was irritated and haemorrhaging present in some animals. These mice also exhibited pale livers and kidneys. The surviving females were normal in appearance at autopsy. Under the conditions of the study the approximate LD50 of the test material was determined to be between 2060 and 5015 mg/kg bw. The study was assigned a reliability score of 1 in line with the principles for assessing data quality as defined by Klimisch (1997).

 

Supporting information is available in the form of a company website (Good Scents Company, 2011), a material safety data sheet (Sigma Aldrich, 2009), and other company data (Moreno, 1973). All acute toxicity data was taken from abstracts of information with no or few details reported on the methodology applied. All three sources report the acute oral toxicity of the test material to be > 5000 mg/kg bodyweight. As such all three studies were assigned a reliability score of 4 in accordance with the principles for assessing data quality defined by Klimisch (1997).

Inhalation

In accordance with point 8.5.2, Column 2 (Specific rules for adaptation from Column 1), Annex VIII of Regulation (EC) No. 1907/2006, an acute inhalation study does not need to be performed as the substance has a low vapour pressure and the use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be unlikely to occur. The acute toxicity endpoint has been addressed by assessing the toxicity via the oral and dermal routes, which is more appropriate when considering the properties of this substance.

Dermal

In the key study (Sanders, 2012), the acute dermal toxicity of the test material was determined in a study conducted in compliance with GLP and the standardised guidelines OECD 402 and EU Method B.3. During the study, two Wistar rats (one male and one female) were given a single, 24 –hour, semi-occluded dermal application of the undiluted test material to intact skin at a dose level of 2000 mg/kg bw. Based on the results of the initial test, a further group of four males and four females was similarly treated. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy at termination. Under the conditions of the study, there were no deaths and no overt signs of systemic toxicity. There were no signs of dermal irritation and all animals showed expected gains in bodyweight over the study period. At necropsy a cavity was noted in the right kidney of one male but this was considered to be a genetic defect and not related to treatment with the test material. No abnormalities were noted at necropsy of the remaining animals. In consideration of the findings, the acute dermal LD50 was determined to be > 2000 mg/kg. The study was assigned a reliability score of 1 in line with the principles for assessing data quality as defined by Klimisch (1997).

Supporting information is available in the form of a company website (Good Scents Company, 2011), a material safety data sheet (Sigma Aldrich, 2009), and other company data (Moreno, 1973). All supporting acute toxicity data was taken from abstracts of information with no or few details reported on the methodology applied. All three sources report the acute dermal toxicity of the test material to be > 5000 mg/kg bw. As such, all three studies were assigned a reliability score of 4 in accordance with Klimisch (1997).


Justification for selection of acute toxicity – oral endpoint
The key study was conducted to sound scientific principles with a sufficient level of reporting to assess the quality of the data submitted. Accordingly the study was assigned a reliability score of 2 according to the criteria of assessing data quality as defined by Klimisch (1997).

Justification for selection of acute toxicity – inhalation endpoint
Data waiver has been submitted to address acute toxicity via the inhalation route.

Justification for selection of acute toxicity – dermal endpoint
Study was conducted to GLP and standardised guidelines with a high level of reporting and was assigned a reliability score of 1 according to the criteria of Klimisch.

Justification for classification or non-classification

According to the criteria outlined in Regulation (EC) No. 1272/2008 and Directive 67/548/EEC, the substance does not meet the criteria for classification for acute oral or dermal toxicity.