Fatty acids, C18 (saturated and unsaturated) ethyl esters

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Two genetic toxicity studies were performed, both followed GLP. Key genetic toxicity in vitro (aberration test and Ames test). No mutagenic effects observed in these tests for the read-across substance CAS No 67762-38-3 (see attached read-across justification document in section 13). A supportive genetic toxicity study (Ames test) was performed with Lipex SheaLight (Fatty acids, C18 (saturated and unsaturated) ethyl esters). No mutagenic activity was seen.

Further, data from an In Vitro Mammalian Cell Gene Mutation Test and an in vivo Mammalian Bone Marrow Chromosome Aberration Test using the read-across substance CAS No 67762-38-3 was negative.  

Data form the read-across substance (CAS No 67762-38-3) as well as data from Fatty acids, C18 (saturated and unsaturated) ethyl esters indicate no mutagenic activity.

Link to relevant study records

Reference

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From 2000-03-23 To 2000-10-03

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was performed according to OECD guideline 473 and GLP on the read-across substance CAS 67762-38-3..

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

lymphocytes:

Details on mammalian cell type (if applicable):

- Type and identity of media: RPMI 1640 containing 20% fetal calf serum, L-glutamine (2mM), penicillin (100 U/ml), streptomycin (100µg/ml) and phytohaemagglutinin.

Metabolic activation:

with and without

Metabolic activation system:

S9 from induced Aroclor 1254 rat liver

Test concentrations with justification for top dose:

- 18.96, 37.93, 75.85, 151.70, 303.41, 606.82, 1213.64 and 2427.27 µg/ml (in the first experiment)
- 75.85, 151.70, 303.41, 606.82, 1213.64 and 2427.27 µg/ml (in the second experiment)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: no justification in the study report

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: mitomycin C without S9 mix (3µg/ml for 3 hours of treatment, 0.2 µg/ml for continuous treatment), cyclophosphamide with S9 mix (50 µg/ml)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION (first experiment)
- Exposure duration: 3h
- Expression time (cells in growth medium): 17h
- Fixation time (start of exposure up to fixation or harvest of cells): 20h

DURATION (second experiment)
- Exposure duration: 3h (without S9); 20h and 44h (with S9)
- Expression time (cells in growth medium): 17h and 41h (without S9); 20h and 44h (with S9)
- Fixation time (start of exposure up to fixation or harvest of cells): 20h and 44h (with and without S9)

STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: approximately 1.5 normal cell cycles or approximately 1.5 normal cell cycles and 24 hours later.

NUMBER OF CELLS EVALUATED: 100 metaphase/culture

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreplication: yes
- Other: gaps, chromatid and chromosome breaks and exchanges, multipleaberrations and pulverization

OTHER: blind scoring

Evaluation criteria:

A reproducible and statistically significant increase in the frequency of cells with structural aberrations for at least one of the dose-levels and one of
the two harvest times was considered as a positive result. Reference to historical data or other considerations of biological relevance, was also taken into account in the evaluation of the findings.

Statistics:

The comparison was performed using the Chi-square test (p = 0.05)

Species / strain:

lymphocytes: primary culture

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

Cytotoxicity:
Without S9:
After 3-hour treatment, a slight decrease in the mitotic index was noted at dose-levels of 1213.64 and 2427.27 µg/ml (up to 34%).
After 20- hour treatment, a 35% decrease in the mitotic index was induced at 2427.27 µg/ml.
After 44-hour treatment, a 33-69% decrease in the mitotic index was noted at dose-levels of 1213.64 and 2427.27 µg/ml.

With S9 mix:
No noteworthy decrease in the mitotic index was noted at the 20 hour harvest time

Chromosal aberration analyses:
The test substance did not induce any significant increase in the frequency of cells with chromosome aberrations in both experiments and at both harvest times, with and without S9 mix.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

Under the experimental conditions, the test substance Esterol C does not induce chromosome aberrations in cultured human lymphocytes.

Executive summary:

In a GLP mammalian cell cytogenetic assay (chromosome aberration) (Haddouk, 2000), primary lymphocyte cultures were exposed to Esterol C (batch no. 0006503) at concentration of 18.96, 37.93, 75.85, 151.70, 303.41, 606.82, 1213.64 and 2427.27 µg/ml with and without metabolic activation. Positive controls induced the appropriate response. There was no evidence of chromosome aberration induced over background.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

In a GLP reverse gene mutation assay in bacteria (Haddouk, 1999) strains of Salmonella typhimurium (TA 1535, TA 1537, TA 98, TA 100 and TA 102) were exposed to Esterol C (CAS No 67762 -38-3) at concentration of, 0, 62.5, 125, 250, 500 and 1000 µg/plate in the presence and absence of mammalian metabolic activation. The positive controls induced the appropriate responses in the corresponding strains. No noteworthy increase in the number of revertants was induced in all tested strains with and without metabolic activation.

This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.

 

In a GLP mammalian cell cytogenetic assay (chromosome aberration) (Haddouk, 2000), primary lymphocyte cultures were exposed to Esterol C (CAS No 67762 -38-3) at concentration of 18.96, 37.93, 75.85, 151.70, 303.41, 606.82, 1213.64 and 2427.27 µg/ml with and without metabolic activation. Positive controls induced the appropriate response. There was no evidence of chromosome aberration induced over background.

This study is classified as acceptable. This study satisfies the requirement for Test Guideline OECD 473 for in vitro mutagenicity (in vitro chromosome aberrations) data.

A Salmonella typhimurium revere mutation standard plate incorporation study (OECD 471) was conducted to evaluate whether Lipex SheaLight (Fatty acids, C18 (saturated and unsaturated) ethyl esters) would cause mutagenic changes in the average number of revertants for histidine dependent Salmonella typhimurium strains TA97a, TA98, TA100, TA102, and TA1535 in the presence and absence of S9 metabolic activation.

Under the conditions used the test article solution was considered to be non-mutagenic to Salmonella typhimurium tester strains TA97a, TA98, TA100, TA102, and TA1535. The negative and positive controls performed as anticipated indicating validity of the study design.

Further, data from an In Vitro Mammalian Cell Gene Mutation Test and an in vivo Mammalian Bone Marrow Chromosome Aberration Test using the read-across substance CAS No 67762-38-3 was negative.

Justification for selection of genetic toxicity endpoint

Esterol C (CAS No 67762-38-3) does not show any mutagenic activity in the bacterial reverse mutation test on Salmonella  typhimurium strains.

Under the experimental conditions, the test substance Esterol C (CAS No 67762-38-3) does not induce chromosome aberrations in cultured human lymphocytes.

Justification for classification or non-classification

From the available data on genetic toxicity for the read-across substance CAS No 67762-38-3 (see attached waiving and justification document in section 13) and for Lipex SheaLight (Fatty acids, C18 (saturated and unsaturated) ethyl esters), no classification apply for Fatty acids, C18 (saturated and unsaturated) ethyl esters).