Squalene-rich fraction obtained from vegetable oil deodorizer distillate by transesterification, crystallisation and vacuum distillation

Endpoint summary

Description of key information

Overall weight of evidence from different studies suggests that the main constituents of the test substance ‘squalene-rich fraction obtained from vegetable oil deodorizer distillate by transesterification, crystallisation and vacuum distillation’ have low acute toxicity following oral and dermal exposure, with LD50 values greater than 2,000 mg/kg bw. This is in line with their long history of safe use in a wide range of nutritional (food and feed), cosmetic and/or industrial applications. The exposure via the inhalation route is not expected given the low vapour pressure and intermediate use of the substance. Therefore, acute inhalation exposure is not expected to pose an issue for human health and no further testing is required for this endpoint according to Annex VIII, Section 8.5, column 2 of the REACH legislation.

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:

July 1984 to December 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

not specified

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: TNO Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann GmbH, Lage, Germany
- Weight at study initiation:
* Males: 170 d on Day -1, 159 g on day of administration
* Females: 151 g on Day -1, 141 g on day of administration
- Fasting period before study: From 16 h pre-dose to 3 h post-dose
- Housing: Macrolon Type 3 cages
- Diet (e.g. ad libitum): Altromin-Haltungsdiät 1324, Fa. Altromin GmbH, 4937 Lage, Germany
- Water (e.g. ad libitum): tap water
- Acclimation period: 6 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 21°C
- Humidity (%): ca. 51%
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

arachis oil

Doses:

5,000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations and weighing: Mortality and clinical signs: sevaral times on the day of administration, twice daily after that; bodyweight once pre-dose, once on day of adminsitration, then after 48 h, 7 and 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, macroscopic observation of organs

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

None

Clinical signs:

Rough fur in males and females 10 min to 1 h after dosing.
No other symptoms.

Body weight:

- Bodyweight gain in males: 67 g at Day 14 post-application
- Bodyweight gain in females: 26 g at Day 14 post-application

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the LD50 of the constituent in rats was determined to be greater than 5,000 mg/kg bw.

Executive summary:

A study was conducted to determine the acute oral toxicity of the constituent 'fatty acids, C16 -18 and C18 -unsatd.' to rats, according to EU Method B1. No treatment-related effects were noted at 5,000 mg/kg bw. Under the conditions of the study, the LD50 of the constituent in rats was therefore considered to be >5,000 mg/kg (Potokar, 1984).

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

August 1984 to December 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

not specified

GLP compliance:

yes

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

other: TNO Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann GmbH, Lage, Germany
- Weight at study initiation:
* Males: 170 d on Day -1, 159 g on day of administration
* Females: 151 g on Day -1, 141 g on day of administration
- Fasting period before study: From 16 h pre-dose to 3 h post-dose
- Housing: Macrolon Type 3 cages
- Diet (e.g. ad libitum): Altromin-Haltungsdiät 1324, Fa. Altromin GmbH, 4937 Lage, Germany
- Water (e.g. ad libitum): tap water
- Acclimation period: 6 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ca. 21°C
- Humidity (%): ca. 51%
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: gavage

Vehicle:

arachis oil

Doses:

5,000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations and weighing: Mortality and clinical signs: sevaral times on the day of administration, twice daily after that; bodyweight once pre-dose, once on day of adminsitration, then after 48 h, 7 and 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, macroscopic observation of organs

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

None

Clinical signs:

No clinical signs observed

Body weight:

- Bodyweight gain in males: 69 g at Day 14 post-application
- Bodyweight gain in females: 21 g at Day 14 post-application

Gross pathology:

No pathological signs

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the acute oral LD50 of the constituent in rats was determined to be greater than 5,000 mg/kg bw.

Executive summary:

A study was conducted to determine the acute oral toxicity of the constituent 'fatty acids, C8 -18 and C18 -unsatd.' in rats, according to EU Method B1. No treatment-related effects were noted at 5,000 mg/kg bw. Under the study conditions, the LD50 of the constituent in rats was considered to be >5,000 mg/kg (Potokar, 1984).

Reference 3

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

GLP compliance:

yes

Remarks:

U.S. Fed. Reg., Titel 21, Dec. 22nd, 1978, part II

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Firma Charles River Wiga, Sandhofer Weg 7, 8714 Sulzfe1d
- Weight at study initiation: Male: 176.4 - 188.9 g; Female: 151.4 - 158.1 g
- Fasting period before study: Yes, 16 h before administartion of the substance
- Housing: Collective caging in macrolon type III/max. 5
- Diet: Ssniff-R Alleindiat flilr Ratten feed, ad libitum
- Water (e.g. ad libitum): Macrolon dringking bottles, Becker & Co. 4620 Castrop-Rauxel, ad libitum
- Acclimation period: 7 d


ENVIRONMENTAL CONDITIONS
- Temperature: 20±2°C
- Humidity: 50-80%
- Air changes (per hr):
- Photoperiod: 12 h dark/12 h light


IN-LIFE DATES: From: Feb. 19, 1988 To: March 3, 1988

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

VEHICLE
- Concentration in vehicle: Undiluted


MAXIMUM DOSE VOLUME APPLIED: 0.95 mL

Doses:

5 mL/kg

No. of animals per sex per dose:

5 male/female

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations and weighing: Clinical observations: About 20 min, 1-2 h, 3-6 h, 24/48 h, Days 3- 6 and 7-14; Body weight: At Days 0 (beginning of the experiment), 7 and 14; Necropsy: Immediately after death of animal and after sacrifice, at the end of the 14 d observation period all surviving
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs, body weight, gross pathology, mortality

Statistics:

Not reported

Preliminary study:

Not applicable

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 mL/kg bw

Remarks on result:

other: equivalent to 4952 mg/kg

Mortality:

No mortalities were observed

Clinical signs:

No clinical symptoms were observed

Body weight:

Body weight changes after the observation period showed a normal weight gain

Gross pathology:

No macroscopic findings in the cranial, thoracic and abdominal cavity

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the acute oral LD50 of the constituent is considered to be >5 mL/kg (equivalent to 4,952 mg/kg).

Executive summary:

A study was conducted to evaluate the acute oral toxicity of the constituent ‘glycerides, C16 and C18-unsatd. and C18-unsatd. hydroxy’ (as castor oil) in rats in accordance with OECD Guideline 401, 1987 and to GefStoffV, Aug. 26th, 1986, (BGBI 1470). A group of 5 fasted male and female Wistar rats were exposed to a single oral dose of 5 mL/kg bw. Clinical signs and mortality were recorded during the 14 day observation period. Necropsy was conducted on moribund animals and those terminated at test end. No clinical signs and no mortality were observed. There were no treatment-related effects on body weight and nothing abnormal was found in the animals necropsied on Day 14. Under the study conditions, the acute oral LD₅₀ of the constituent was considered to be >5 mL/kg (equivalent to 4,952 mg/kg) (Chibanguza, 1988).

Reference 4

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

no guideline followed

Principles of method if other than guideline:

In an acute oral toxicity study, palm oil was administered at a single dose of 5,000 mg/kg to 5 rats, followed of observation of the mortality and clinical signs.

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Species:

rat

Route of administration:

oral: gavage

Doses:

5000 mg/kg

No. of animals per sex per dose:

5 animals

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Mortality:

Two mortalities were reported on Day 2 and 6

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the acute oral LD50 of the constituent in rats was found to be greater than 5000 mg/kg.

Executive summary:

An acute oral toxicity was conducted with the constituent glycerides, C16 -18 and C18 -unsatd. (palm oil) according to OECD guideline 401 and EU method B.1. The constituent was administered at a dose of 5,000 mg/kg to 5 rats. Two moratlities were reported on Day 2 and 6. Therefore, under the study conditions, the acute oral LD50 of the constituent in rats was considered to be greater than 5,000 mg/kg (CTFA, 1978).

Reference 5

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

February 15, 1988 to March 03, 1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

not specified

Test type:

standard acute method

Limit test:

yes

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Firma Charles River Wiga, Sandhofer Weg 7, 8714 Sulzfeld
- Weight at study initiation: male: 191.2 - 197.5 g and female: 170.8 - 185.6 g
- Fasting before study: Yes
- Housing: Collective caging, cage type: Macrolon type III
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 7 d


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 22°C
- Humidity (%): 50 - 80%
- Photoperiod: Fluorescent light, 120 lux, 12 h/d

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Doses:

4763 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 d
- Frequency of observations and weighing: Recorded on Days 0, 7 and 14
- Necropsy of survivors performed: Yes
- Other examinations performed: Clinical signs, organ weights, histopathology, CNS symptoms, coordination, tone, reflex, autonomic functions

Preliminary study:

Two female rats were employed in a preliminary range finding study. The dose of the single oral administration was 5 mL/kg bw.

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 4 763 mg/kg bw

Mortality:

No mortalities were observed

Clinical signs:

No clinical signs were observed

Body weight:

Normal weight gain

Gross pathology:

No macroscopic findings in the cranial, thoracic and abdominal cavity

Interpretation of results:

GHS criteria not met

Conclusions:

Under the study conditions, the acute oral LD50 of the constituent in rats was found to be greater than 4,763 mg/kg.

Executive summary:

The acute oral toxicity of ‘glycerides, C16-18 and C18-unsatd.’ (as linseed oil) was investigated in rats in accordance with OECD Guidenile 401 and EG 84/449/EWG.

A single dose of 4,763 mg/kg bw of the test substance was administered to 5 male and 5 female rats. Clinical signs, body weights and mortalities were recorded during the 14 d observation period. Immediately after death or at the end of the observation period, a complete necropsy was performed. No mortality or any other adverse effect was observed in any of the animals. Therefore the oral LD50 of the test substance in rats was determined to be greater than 4,763 mg/kg (Chibanguza, 1988).

Reference 6

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Qualifier:

no guideline followed

Principles of method if other than guideline:

Method followed unknown, data from handbook

GLP compliance:

not specified

Species:

rat

Route of administration:

oral: unspecified

Doses:

up to 5000 mg/kg bw

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 of the constituent in rats is greater than 5,000 mg/kg bw.

Executive summary:

A study was conducted to determine the acute oral toxicity potential of the constituent glycerides, C8 -18, and C18 -unstad. (coconut oil) according to an unspecified method. Under the study conditions, the oral LD50 of 'glycerides, C8 -18 and C18-unsatd.' (as coconut oil) in rats is greater than 5,000 mg/kg bw (Biotech Index, 1970).

Reference 7

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Qualifier:

no guideline followed

Principles of method if other than guideline:

Single oral dose of coconut oil was administered to rats to determine the acute oral toxicity potential.

GLP compliance:

not specified

Limit test:

yes

Species:

rat

Route of administration:

oral: unspecified

Doses:

26 mL/kg bw (23,500 mg/kg bw)

No. of animals per sex per dose:

10 animals

Control animals:

not specified

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 23 500 mg/kg bw

Mortality:

No mortality

Interpretation of results:

GHS criteria not met

Conclusions:

The oral LD50 of the coconut oil in rats is greater than 23,500 mg/kg bw.

Executive summary:

A study was conducted to determine the acute toxicity potential of the constituent glycerides, C8 -18 and C18 -unsatd. (coconut oil) in rats. In this study, 26 mL/kg bw (23,500 mg/kg bw) of ‘glycerides, C8-18 and C18-unsatd.’ (as coconut oil) was orally administered to 10 rats. No mortality was observed. Under the test conditions, the oral LD50 of the constituent in rats can be considered to be greater than 23,500 mg/kg bw (IUCLID, 2000).

Reference 8

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Acute oral toxicity of the constituent squalene was investigated during the seven-day observation period, by administering undiluted doses of 5.0, 12.5, 25.0, and 50.0 mL/kg to groups of 5, 5, 10, and 10 mice respectively.

GLP compliance:

not specified

Species:

mouse

Route of administration:

oral: unspecified

Doses:

5.0, 12.5, 25.0, and 50.0 mL/kg (equivalent to 4,300, 10,750, 21,500 and 43,000 mg/kg respectively, assuming the density of squalene as 0.86 g/mL)

No. of animals per sex per dose:

5.0 mL/kg: 5 mice
12.5 mL/kg: 5 mice
25.0 mL/kg: 10 mice
50.0 mL/kg: 10 mice

Control animals:

not specified

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 50 mL/kg bw

Based on:

test mat.

Remarks on result:

other: equivalent to >43,000 mg/kg bw

No toxic effects were observed and no deaths occurred during the seven day observation period.

Interpretation of results:

other: EU CLP criteria not met

Conclusions:

Under the test conditions, the acute oral LD50 of the constituent in mice was found to be >50 mL/kg (equivalent to 43,000 mg/kg).

Executive summary:

A multiple dose study was conducted to investigate the acute oral toxicity of the constituent squalene in mice. Groups of 5, 5, 10, and 10 mice received undiluted doses of 5.0, 12.5, 25.0, and 50.0 mL/kg (equivalent to 4,300; 10,750; 21,500 and 43,000 mg/kg bw respectively of the test substance, assuming a density of 0.86 g/mL). The effects were evaluated during a period of seven days. No toxic effects were observed and no deaths occurred during the seven day observation period. Under the test conditions, the acute oral LD50 of the substance in mice was found to be >50 mL/kg (equivalent to 43,000 mg/kg) (CIR, 1982).

Reference 9

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Method followed unknown, data from SCF opinion

GLP compliance:

not specified

Species:

rat

Route of administration:

oral: unspecified

Preliminary study:

No data

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the acute oral LD50 of the constituents in rats was found to be greater than 2,000 mg/kg.

Executive summary:

A study was conducted to determine the acute toxicity potential of the constituent by an unspecified method. Under the test conditions, the acute oral LD50 of the constituent β-sitosterol in rats was foudn to be greater than 2,000 mg/kg (SCF, 2003).

Reference 10

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The oral LD50 of undiluted tocopherol was determined using groups of five male ddY mice.

GLP compliance:

not specified

Species:

mouse

Strain:

other: ddY

Sex:

male

Route of administration:

oral: unspecified

Doses:

10 mL/kg (equivalent to 23,812.5 mg/kg, assuming density of tocopherol as 0.9525 g/mL)

No. of animals per sex per dose:

5 male mice

Control animals:

not specified

Statistics:

LD50 was calculated using probit analysis

Key result

Sex:

male

Dose descriptor:

LD50

Effect level:

25 mL/kg bw

Based on:

test mat.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the acute oral LD50 of the constituent was found to be greater than 25 mL/kg bw (equivalent to 23,813 mg/kg).

Executive summary:

A study was conducted to determine the acute toxicity potential of the constituent by an unspecified method. Five male ddY mice were administered a single dose at a volume of 10 mL/kg ( (equivalent to 23,813 mg/kg, assuming density of the constituent as 0.9525 g/mL) and then observed for 14 d. Under the test conditions, the acute oral LD50 of the undiluted constituent was found to be >25 mL/kg bw (23,813 mg/kg) (Fiume, 2002).

Reference 11

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The acute oral LD50 value for d-alpha-tocopheryl succinate was determined in young adult Charles River CD rats of both sexes.

GLP compliance:

not specified

Species:

rat

Strain:

other: Charles River CD

Sex:

male/female

Route of administration:

oral: unspecified

Control animals:

not specified

Key result

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 7 000 mg/kg bw

Based on:

test mat.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the acute oral LD50 of the constituent was found to be >7,000 mg/kg in Charles River CD rats.

Executive summary:

A study was conducted to determine the acute toxicity potential of the constituent by an unspecified method. Under the test conditions, the acute oral LD50 of the constitute d-alpha-tocopheryl succinate, as cited in the JECFA evaluation report, was >7,000 mg/kg in Charles River CD rats (JECFA, 1987).

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

All studies with constituents were taken from peer reviewed or regulatory opinions

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Qualifier:

no guideline followed

Principles of method if other than guideline:

Single dose of fully hydrogenated coconut oil was applied dermally to guinea pigs and animals were observed for 7 d.

GLP compliance:

not specified

Limit test:

yes

Species:

guinea pig

Type of coverage:

not specified

Duration of exposure:

Single dermal application

Doses:

3,000 mg/kg

No. of animals per sex per dose:

12

Control animals:

not specified

Key result

Sex:

not specified

Dose descriptor:

LD0

Effect level:

3 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality during 7 d observation period

Interpretation of results:

GHS criteria not met

Conclusions:

Under test conditions, the acute dermal LD0 of the substance in guinea pigs was found to be greater than 3,000 mg/kg.

Executive summary:

A study was conducted to determine the acute dermal toxicity of the constituent glycerides, C8 -18 and C18 -unsatd. (in the form of fully hydrogenated coconut oil) in guinea pigs. The substance was applied at a dose of 3,000 mg/kg to the skin of 12 guinea pigs. No mortality occurred during the 7 d observation period. Under test conditions, the LD0 of the substance in guinea pigs was found to be 3,000 mg/kg (CIR, 1986).

Reference 2

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

Method followed unknown, acute dermal LD50 value of tocopheryl acetate in rats was cited in the CIR review report

GLP compliance:

not specified

Species:

rat

Type of coverage:

not specified

Key result

Sex:

not specified

Dose descriptor:

LD50

Effect level:

> 3 000 mg/kg bw

Based on:

test mat.

Interpretation of results:

GHS criteria not met

Conclusions:

Under the test conditions, the acute dermal LD50 value of the constituent in rats was found to be >3,000 mg/kg.

Executive summary:

A study was conducted to determine the acute dermal toxicity potential of the constitutent tocopherol by an unspecified method. Under the test conditions, the acute dermal LD50 of the substance in rats was found to be >3,000 mg/kg (Fiume, 2002).

Reference 3

Endpoint:

acute toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the physicochemical and toxicological properties suggest no potential for a significant rate of absorption through the skin

the study does not need to be conducted because the substance does not meet the criteria for classification as acute toxicity or STOT SE by the oral route and no systemic effects have been observed in in vivo studies with dermal exposure (e.g. skin irritation, skin sensitisation)

Reason / purpose for cross-reference:

data waiving: supporting information

Reason / purpose for cross-reference:

data waiving: supporting information

Reason / purpose for cross-reference:

data waiving: supporting information

Reason / purpose for cross-reference:

data waiving: supporting information

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

All studies with constituents were taken from peer reviewed or regulatory opinions.

Additional information

In the absence of acute toxicity study with the test substance, ‘squalene-rich fraction obtained from vegetable oil deodorizer distillate by transesterification, crystallisation and vacuum distillation’ the endpoint has been assessed based on studies for substances representative of the main constituents, which can be categorised as glycerides, fatty acids or fatty acid methyl esters (which will eventually hydrolyse to fatty acids (Mattson and Volpenhein, 1972)) and unsaponifiable matter (including tocopherols, sterols, squalene and hydrocarbons). Asa large number of studies have been conducted on the individual constituents, particularly in the context of nutritional research, for practical reasons, only a limited number of studies are reported below:

 

Oral:

Glycerides:

A study was conducted to evaluate the acute oral toxicity of the constituent ‘glycerides, C8-18 and C18-unsatd.’ (as coconut oil) in rats. In this study, 26 mL/kg (equivalent to 23,500 mg/kg bw) of the substance was orally administered to 10 rats. No adverse effects or mortality was observed and the oral LD50 was considered to be greater than 23,500 mg/kg (IUCLID dataset, 2000). This is further supported by the acute oral LD50 of 5000 mg/kg identified for the constituent 'glycerides, C8 -18 and C18-unsatd.' (as coconut oil) in the Biotech Index data (Biotech Index, 1970).

Similar results were obtained in several acute toxicity studies conducted with different vegetable oil constituents containing higher chain glycerides, C16 and C18 -unsatd. in rats. One of the studies was conducted to evaluate the acute oral toxicity of the constituent ‘glycerides, C16 and C18 -unsatd. and C18 -unsatd. hydroxy’ (as castor oil) in rats in accordance with OECD Guideline 401, 1987 and to GefStoffV, Aug. 26th, 1986, (BGBI 1470). In this study, a group of 5 fasted male and female Wistar rats were exposed to a single oral dose of 5 mL/kg followed by routine monitoring of all the parameters. There were no treatment-related effects and the acute oral LD50 of the substance was therefore considered to be >5 mL/kg (equivalent to 4,952 mg/kg) (Chibanguza, 1988a).

Another study was conducted to evaluate the acute oral toxicity of the constituent ‘glycerides, C16 -18 and C18 -unsatd.’ (as linseed oil) in rats in accordance with OECD Guideline 401 and EG 84/449/EWG. No mortality or other adverse effects were observed following oral administration of single dose of 4,763 mg/kg to 5 male and 5 female rats. Therefore, the acute oral LD50 of the substance in rats was determined to be greater than 4,763 mg/kg bw (Chibanguza, 1988b).

Similar results were obtained in another study conducted with the constituent palm oil containing ‘glycerides, C16-18 and C18-unsatd.’ in rats. In this study, out of 5 dosed animals two mortalities were reported on Day 1 and 6 post intubation of 5,000 mg/kg of the substance. Therefore, under the conditions of the study the oral LD50 of the substance in rats was considered to be greater than 5,000 mg/kg (CIR, 2000).

Fatty acids:

Two studies were conducted to determine the acute oral toxicity of the constituents 'fatty acids, C8-18 and C18 -unsatd.' and 'fatty acids, C16 -18 and C18 -unsatd.' to rats in accordance with EU Method B1. No treatment-related effects were noted in any of the studies at the only tested dose of 5,000 mg/kg. Under the conditions of the studies, the acute oral LD50 of the substances in rats was therefore considered to be > 5,000 mg/kg (Potokar, 1984).

Fatty acids methyl esters

Additional data for fatty acid Me ester, not needed as they will hydrolyse to fatty acids which are being addressed (Mattson and Volpenhein, 1972).

Unsaponifiable matter:

Tocopherols:

A study was conducted to determine the acute toxicity potential of the constituent by an unspecified method. Under the test conditions, the acute oral LD50 of the constitute d-alpha-tocopheryl succinate, as cited in the JECFA evaluation report, was >7,000 mg/kg in Charles River CD rats (JECFA, 1987).

A study was conducted to determine the acute toxicity potential of the constituent by an unspecified method. Five male ddY mice were administered a single dose at a volume of 10 mL/kg (equivalent to 23,813 mg/kg, assuming density of the constituent as 0.9525 g/mL) and then observed for 14 d. Under the test conditions, the acute oral LD50 of the undiluted constituent was found to be >25 mL/kg bw (23,813 mg/kg) (Fiume, 2002).

Sterols:

A study was conducted to determine the acute toxicity potential of the constituent by an unspecified method. Under the test conditions, the acute oral LD50 of the constituent β-sitosterol in rats was found to be greater than 2,000 mg/kg (SCF, 2003).

Squalene:

A multiple dose study was conducted to investigate the acute oral toxicity of the constituent squalene in mice. Groups of 5, 5, 10, and 10 mice received undiluted doses of 5.0, 12.5, 25.0, and 50.0 mL/kg (equivalent to 4,300; 10,750; 21,500 and 43,000 mg/kg bw respectively of the test substance, assuming a density of 0.86 g/mL). The effects were evaluated during a period of seven days. No toxic effects were observed, and no deaths occurred during the seven-day observation period. Under the test conditions, the acute oral LD50 of the substance in mice was found to be >50 mL/kg (i.e., equivalent to 43,000 mg/kg) (CIR, 1982)

Hydrocarbon:

Acute oral toxicity studies with white mineral oil (C15-C50, highly refined) or light paraffinic distillate (C15-C30, viscosity <19 mm2/s) revealed low acute oral toxicity with LD50 value >5000 mg/kg bw in SD rats (EFSA, 2012). Further, a review article by Mckee et al (2015) reported that, the constituents of C14–C20 aliphatic (<2% aromatic) hydrocarbon solvents have limited potential to produce acute toxicity. Unpublished data from acute oral toxicity tests indicate that the LD50 values are greater than the highest levels tested (oral LD50: >5000 mg/kg bw) (Mckee, 2015).

Overall weight of evidence from different studies suggests that the main constituents of the test substance 'squalene-rich fraction obtained from vegetable oil deodorizer distillate by transesterification, crystallisation and vacuum distillation’ have low acute toxicity following oral and dermal exposure, with LD50 values greater than 2,000 mg/kg bw. This is in line with their long history of safe use in a wide range of nutritional (food and feed), cosmetic and/or industrial applications.

Inhalation:

Exposure via the inhalation route is not expected considering the intermediate use and low vapour pressure (0.00108 Pa at 20°C) of the substance. In addition, studies conducted via the oral and dermal route with the constituents or its representative substances indicated a lack of significant toxicity of the test substance. Therefore, testing via inhalation route is unlikely to result in any additional hazard identification and hence further testing involving vertebrate animals may be omitted, in accordance with Annex XI (1.2) and Annex VIII, Section 8.5, column 2 of the REACH regulation.

Dermal:

Glycerides:

A study was conducted to evaluate the acute dermal toxicity of the constituent 'glycerides, C8 -18 and C18 -unsatd.' (in the form of fully hydrogenated coconut oil) in guinea pigs. In this study, the undiluted substance was applied at a dose of 3,000 mg/kg to the skin of 12 guinea pigs. No deaths occurred during the 7-d observation period. Under test conditions, the LD0 of the substance was determined to be 3,000 mg/kg.

Fatty acids:

According to the reviews conducted in the context of the HERA project, fatty acids with chain lengths varying between C10-18, including C18-unsatd. show low acute dermal toxicity, with LD50 values above 2,000 mg/kg (HERA, 2002; CIR, 1987).

Fatty acid methyl esters:

Additional data for fatty acid Me ester, not needed as they will hydrolyse to fatty acids which are being addressed (Mattson and Volpenhein, 1972).

Unsaponifiable matters:

Tocopherols:

The acute dermal LD50of the constituent tocopheryl acetate was found to be greater than 3,000 mg/kg in a review by the CIR panel (2002).

Sterol and sterol esters:

Sterols and sterol esters are generally poorly absorbed via skin so that no systemic toxicity from dermal exposure is expected.

Squalene:

Due to low acute oral toxicity, low dermal absorption and absence of systemic effects in skin sensitisation studies, squalene is expected to have low acute dermal toxicity and no additional testing is needed as per VIII, Section 8.5.3, column 2 of the REACH regulation.

Hydrocarbons:

Hydrocarbon are generally poorly absorbed via skin so that no systemic toxicity from dermal exposure is expected. In a review by the CIR panel, the acute dermal LD50 of the isoparaffin were found to be greater than 3200 mg/kg bw in rabbits (2012). Further, a review article by Mckee et al (2015) reported that, the constituents of C14–C20 aliphatic (<2% aromatic) hydrocarbon solvents have limited potential to produce acute toxicity. Unpublished data from acute dermal toxicity tests indicate that the LD50 values are greater than the highest levels tested (dermal LD50: >2000 mg/kg bw) (Mckee, 2015).

 

Overall, the weight of evidence suggests that the test substance ‘squalene-rich fraction obtained from vegetable oil deodorizer distillate by transesterification, crystallisation and vacuum distillation’ is considered to have low acute toxicity via oral, dermal or inhalation routes.

Justification for classification or non-classification

The acute toxicity potential of the test substance test substance ‘squalene-rich fraction obtained from vegetable oil deodorizer distillate by transesterification, crystallisation and vacuum distillation’can be deduced based on information on the relevant individual constituents present at >1% concentrations.   

 

Except for the heptane, studies conducted with various other components suggest that they are not acutely toxic via oral and dermal routes and hence do not warrant any classification according to EU CLP criteria (EC 1272/2008). The constituent heptane has been classified as ‘STOT SE 3’ in the 1^stadaptation to technical progress (ATP) of the Classification, Labelling and Packaging (CLP) Regulation. Therefore, as per the CLP regulation, when determining the classification of mixtures (and UVCBs) containing components that are classified for STOT SE3, the concentrations of the individual components must be compared against the generic concentration limits (GCL) for the respective endpoints.    

Based on the above criteria, and considering the current composition, the test substance ‘squalene-rich fraction obtained from vegetable oil deodorizer distillate by transesterification, crystallisation and vacuum distillation’does not warrant classification for acute toxicity according to EU CLP criteria (EC 1272/2008). ​