Fatty acids, C8-10, C12-18-alkyl esters

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

09 Apr - 10 May 2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

GLP - Guideline study. According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.

Justification for type of information:

see attached read across justification

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

no

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

2-ethylhexyl oleate

EC Number:

247-655-0

EC Name:

2-ethylhexyl oleate

Cas Number:

26399-02-0

Molecular formula:

C26H50O2

IUPAC Name:

2-ethylhexyl octadec-9-enoate

Method

Target gene:

Not applicable

Species / strain

Species / strain / cell type:

lymphocytes: cultured peripheral human lymphocytes

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9-mix

Test concentrations with justification for top dose:

3, 10 and 33 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Ethanol

Controls

Controls 1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

cyclophosphamide

Remarks:

with rat liver S9-mix

Migrated to IUCLID6: 10 µg/mL

Controls 2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

without rat liver S9-mix

Migrated to IUCLID6: 0.5 µg/mL for a 3 h exposure period, 0.2 µg/mL for a 24 h exposure period and 0.1 µg/mL for a 48 h exposure period

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 3, 24 and 48h
- Fixation time (start of exposure up to fixation or harvest of cells): 3h treatment: 24 and 48h; 24h treatment: 24h; 48h treatment: 48h

SPINDLE INHIBITOR (cytogenetic assays): colchicine 0.5 µL/mL medium
STAIN (for cytogenetic assays): Giemsa 5% (v/v) in tap water

NUMBER OF REPLICATIONS: 2 (duplicates at the 3h-exposure time)

NUMBER OF CELLS EVALUATED: 100 per culture

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of 1000 cells

OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

A test substance was considered positive (clastogenic) in the chromosome aberration test if: a) It induced a dose-related statistically significant (Chi-square test, one-sided, p < 0.05) increase in the number of cells with chromosome aberrations. b) A statistically significant and biologically relevant increase in the frequencies of the number of cells with chromosome aberrations was observed in the absence of a clear dose-response relationship.
A test substance was considered negative (not clastogenic) in the chromosome aberration test if none of the tested concentrations induced a statistically significant (Chi-square test, one-sided, p < 0.05) increase in the number of cells with chromosome aberrations. The preceding criteria are not absolute and other modifying factors might enter into the final evaluation decision.

Statistics:

Chi-square test, one-sided, p < 0.05

Results and discussion

Test results

Species / strain:

lymphocytes: cultured human peripheral lymphocytes

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: At a concentration of 33 µg/mL 2-ethylhexyl oleate precipitated in the culture medium
- Other confounding effects: 2-ethylhexyl oleate showed no significant effects on the number of polyploid cells

RANGE-FINDING/SCREENING STUDIES: The highest concentration analysed was selected based on the solubility of the test substance in the culture medium (3 h and 24 h continuous exposure) or on toxicity, inhibition of the mitotic index of about 50% or greater (48 h continuous exposure).

ADDITIONAL INFORMATION ON CYTOTOXICITY:
No cytotoxicity was observed in the duplicate cultures of the 3h exposure time.

Remarks on result:

other: strain/cell type: cultured human peripheral lymphocytes

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

 Table 1: Cytotoxic and Genotoxic observations

Test item	Concentration	Mitotic Index	Aberrant cells in %
	in µg/mL	in %	with gaps	without gaps
Exposure period 3 h, fixation time 24 h, without S9 mix
Ethanol	1.0% (v/v)	100	2	2
MMC	0.5	67	31	30
Test substance	3	99	1	1
	10	98	2	2
	33	92	1	1
Exposure period 3 h, fixation time 24 h, with S9 mix
Ethanol	1.0% (v/v)	100	1	1
CP	0.5	51	38	38
Test substance	3	101	1	1
	10	108	0	0
	33	103	3	3
Exposure period 24 h, fixation time 24 h, without S9 mix
Ethanol	1.0% (v/v)	100	1	1
MMC	0.2	54	34	33
Test substance	3	99	1	1
	10	103	3	3
	33	70	4	3
Exposure period 48 h, fixation time 48 h, without S9 mix
Ethanol	1.0% (v/v)	100	2	0
MMC	0.1	120	51	49
Test substance	3	108	1	0
	10	100	0	0
	33	99	2	2
Exposure period 3 h, fixation time 48 h, with S9 mix
Ethanol	1.0% (v/v)	100	0	0
CP	10.0	--	44	44
Test substance	3	100	2	2
	10	94	2	2
	33	97	1	0

                    MMC: Mitomycin           CP: Cyclophosphamide

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative