Fatty acids, C18 unsaturated, ethyl & methyl esters

Endpoint summary

Description of key information

Oral (rat): The available acute oral toxicity studies resulted in an acute oral LD50 > 5000 mg/kg bw.
Inhalation (rat): The available acute inhalation toxicity study resulted in an acute inhalation LC50 > 2.02 mg/L bw. 
Dermal (rat): The available acute dermal toxicity study resulted in an acute dermal LD50 > 5000 mg/kg bw. 

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute 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: Hilltop Lab animals, Scottdale, Pennsylvania, USA
- Weight at study initiation: 203 - 213 g (males), 189 - 209 g (females)
- Fasting period before study: approximately 21 h
- Housing: individually housed in suspended stainless steel caging with mesh floors
- Diet: Purina Rodent Chow #5012, ad libitum
- Water: filtered tap water, ad lbitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 21
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Doses:

5000 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 days
- Frequency of observations and weighing: Individual weights were recorded just prior to test substance administration and again on days 7 and 14 (termination), the animals were observed for signs of gross toxicity, behavioral changes and mortality at 1, 3 and 24 h post-dosing and at least once daily thereafter for 14 days.
- Necropsy of survivors performed: yes (tissues of the thoracic and abdominal cavities were examined)

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred during the study period.

Clinical signs:

No clinical signs occurred during the study period.

Body weight:

All animals gained weight.

Gross pathology:

Gross necropsy findings were generally unremarkable.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises adequate, reliable (Klimisch score 1-2) and consistent studies, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study with acceptable restrictions (purity: 76%)

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute 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: Ace Animals, Inc., Boyertown, Pennsylvania, USA
- Age at study initiation: 10 - 11 weeks old
- Weight at study initiation: 325 - 341 g (males), 203 - 243 g (females)
- Housing: individually housed in suspended stainless steel caging with mesh floors
- Diet: Purina Rodent Chow #5012, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 21 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 22
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

whole body

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: rectangular whole body Plexiglas chamber
- Exposure chamber volume: 150 L
- Source and rate of air: air compressor (JUN-AIR), approximately 25.1 L/min filtered air
- System of generating particulates/aerosols: The test atmosphere was generated using a l/4 inch JCO atomizer, FC4 fluid cap and AC 1502 air cap (Spraying Systems Inc.). Compressed air was supplied at 30 psi. The test substance was metered to the atomisation nozzle through Size 14 Master Flex Tygon tubing, using a Master Flex Pump Model 7520-35.
- Temperature, humidity in air chamber: 19-21°C, 37-65%

TEST ATMOSPHERE
- Brief description of analytical method used: Gravimetric samples were withdrawn at 6 intervals from the breathing zone of the animals. Samples were collected using 25 mm glass fiber filters (GF/B Whatman) in a filter holder attached by 1/4 inch tygon tubing to a Reliance Electric vacuum pump Model #G557X. Filter papers were weighed before and after collection to determine the mass collected. This value was divided by the total volume of air sampled to determine the chamber concentration. The collections were carried out for 3 minutes at airflows of 4 L/min. Sample airflows were measured using an Omega Flowmeter Model #FMA 5610.
- Samples taken from breathing zone: yes

TEST ATMOSPHERE
- Particle size distribution: An eight-stage Andersen cascade impactor was used to assess the particle size distribution of the test atmosphere. Samples were withdrawn from the breathing zone of the animals at two intervals. The filter paper collection stages were weighed before and after sampling to determine the mass collected upon each stage.
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.): The aerodynamic mass median diameter and geometric standard deviation were determined graphically using two-cycle logarithmic probit axes.

Analytical verification of test atmosphere concentrations:

yes

Remarks:

gravimetrically

Duration of exposure:

4 h

Remarks on duration:

(also 15 min)

Concentrations:

The gravimetric and nominal chamber concentrations were 2.02 and 13.37 mg/L, respectively.

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Mortality, signs of gross toxicity, and behavioral changes were observed prior to exposure, at least every 30 min during exposure, upon removal from the exposure chamber and at least once daily thereafter for 14 days.
- Necropsy of survivors performed: yes (tissues and organs of the thoracic and abdominal cavities were examined)

Preliminary study:

Prior to initiation of the full inhalation study, pre-test trials were conducted to establish generation procedures to achieve, to the extent possible, the desired chamber concentration (2.0 mg/L) and desired particle size distribution (mass median aerodynamic diameter less than or equal to 4 μm). The exposure procedures and atomisation equipment used were based on the results of pre-test trial which provided a gravimetric concentration of 2.11 mg/L and a mass median aerodynamic diameter of 3.1 μm.

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 2.02 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

No mortality occurred.

Clinical signs:

other: In-chamber animal observations included ocular and nasal discharge, hunched posture and hypoactivity. Upon removal from the exposure chamber, all animals recovered from the above symptoms and appeared active and healthy for the 14-day observation period.

Body weight:

All animals gained weight over the 14-day observation period.

Gross pathology:

No gross abnormalities were noted.

Table 1: Summary of particle size distribution

Sample No.	Time of Sample (hours)	Collection Time (minutes)	Mass Median Aerodynamic Diameter (µm)	Geometric Standard Deviation
1	1.5	2	3.0	1.87
2	3	2	3.0	1.78

 

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP guideline study with acceptable restrictions (purity: 80%)

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Version / remarks:

(1987)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1200 (Acute Dermal Toxicity)

Version / remarks:

(1998)

Deviations:

no

GLP compliance:

yes

Test type:

standard acute 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: Sage Labs (no further details)
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: 252 - 270 g (males), 180 - 200 g (females)
- Fasting period before study: overnight
- Housing: individually housed in suspended stainless steel caging
- Diet: Harlan Teklad Global 16% Protein Rodent Diet #2016, ad libitum
- Water: filtered tap water, ad lbitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 - 23
- Humidity (%): 50 - 58
- Air changes (per hr): 13
- Photoperiod (hrs dark / hrs light): 12 / 12

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: dorsal area and the trunk; clipping on the day prior to application
- % coverage: approximately 10% of the body surface (a dose area of approximately 5 inches x 7.6 cm)
- Type of wrap if used: covered with a gauze pad, the gauze pad and entire trunk of each animal were then wrapped with Durapore tape

REMOVAL OF TEST SUBSTANCE
- Washing: yes, with a 3% soap solution followed by tap water and a clean paper towel
- Time after start of exposure: 24 h

Duration of exposure:

24 h

Doses:

5000 mg/kg bw

No. of animals per sex per dose:

5

Control animals:

not required

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations: mortality, signs of gross toxicity, and behavioral changes during the first several hours after application, after patch removal, and then at least once daily thereafter for 14 days, individual body weights of the animals were recorded prior to test substance application (initial) and again on days 7 and 14.
- Necropsy of survivors performed: yes, at the end of the observation period, all animals were euthanized via carbon dioxide and subjected to necropsy (tissues and organs ofthe thoracic and abdominal cavities were examined)

Statistics:

Statistical analysis was limited to the calculation of the mean density value for dosing.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Mortality:

No mortality occurred.

Clinical signs:

Following application, nine animals exhibited ano-genital staining but recovered from these symptoms by day 3. Aside from dermal irritation noted in one animal on days 11-14, there were no other signs of gross toxicity, dermal irritation, adverse clinical effects, or abnormal behavior.

Body weight:

All animals gained weight during the study.

Gross pathology:

No gross abnormalities were noted for any of the animals when necropsied at the conclusion of the 14-day observation period.

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

CLP: not classified

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study, and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

Additional information

Acute toxicity

There are data available for acute oral, inhalation and dermal toxicity available for Fatty acids, C18 unsaturated, ethyl & methyl esters (EC 941-981-3).

Acute oral toxicity

An acute oral toxicity study (limit test) with EC 941-981-3 according to OECD Guideline 401 was performed (Wnorowski, 1995). Groups of 5 male and female Sprague-Dawley rats received single oral doses of 5000 mg/kg bw/day. No mortalities were observed during the study period. No signs of clinical toxicity were reported. All animals showed the expected gain in body weight. The acute oral LD50 in rats was found to be greater than 5000 mg/kg bw/day.

Additionally, an acute oral toxicity study (limit test) with EC 941-981-3 (purity: 80%) according to OECD Guideline 425 was performed (Lowe, 2015). A group of 3 female Sprague-Dawley rats received single oral doses of 5000 mg/kg bw. No mortalities were observed during the study period. No signs of clinical toxicity were reported. All animals showed the expected gain in body weight. The acute oral LD50 in rats was found to be greater than 5000 mg/kg bw.

 

Acute toxicity following inhalation

An acute inhalation study was performed with EC 941-981-3 (purity: 76%) according to OECD Guideline 403 in groups of 5 male and 5 female Sprague-Dawley rats (Wnorowski, 2003). The animals were exposed to an analytical concentration of 2.02 mg/L of the test substance for 4 hours in a rectangular whole body Plexiglas chamber. An aerosol was generated using a 1/4 inch JCO atomizer, FC4 fluid cap and AC 1502 air cap (Spraying Systems Inc.). No mortalities were reported during the exposure or within the 14 day observation period. In-chamber animal observations included ocular and nasal discharge, hunched posture and hypoactivity. Upon removal from the exposure chamber, all animals recovered from the above mentioned symptoms and appeared active and healthy over the observation period. Additionally, all animals gained weight during the observation period. No abnormalities were found at macroscopic post-mortem examination of the animals. The inhalatory 4 h LC50 value in Sprague-Dawley rats was found to exceed 2.02 mg/L. 

 

Acute dermal toxicity

An acute dermal toxicity (limit test) was performed with EC 941-981-3 (purity 80%) according to OECD Guideline 402 (Lowe, 2015). 5 male and female Sprague-Dawley rats were exposed to 5000 mg test substance /kg bw for 24 h under occlusive conditions. The observation period was 14 days. No mortality was observed. Following application, nine animals exhibited ano-genital staining but recovered from these symptoms by day 3. Aside from dermal irritation noted in one animal on days 11-14, there were no other signs of gross toxicity, dermal irritation, adverse clinical effects, or abnormal behavior. Body weight gain and necropsy at study termination revealed no abnormalities. Thus, the acute dermal LD50 in rats was found to be greater than 5000 mg/kg bw.

Overall conclusion

In summary, reliable data available for Fatty acids, C18 unsaturated, ethyl & methyl esters indicate a very low level of acute toxicity following the oral or dermal route and after inhalation. The derived oral and dermal LD50 values and also the LC50 value were greater than the currently applied limit values. Thus, as the available data did not identify any hazard for acute oral and dermal toxicity as well as for acute toxicity following inhalation Fatty acids, C18 unsaturated, ethyl & methyl esters is not considered as hazardous after acute exposure.

Justification for selection of acute toxicity – oral endpoint
The selected study is the most adequate and reliable study based on overall quality assessment (refer to the endpoint discussion for further details).

Justification for selection of acute toxicity – inhalation endpoint
The reliable GLP compliant OECD Guideline study was taken.

Justification for selection of acute toxicity – dermal endpoint
The reliable GLP compliant OECD Guideline study was taken.

Justification for classification or non-classification

The available data on acute oral, inhalation and dermal toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.