Fatty acids, C14-18 and C16-18-unsatd., esters with propylene glycol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

No discussion of results and no historical control data.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

yes

Remarks:

no discussion of results and no historical control data.

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

no discussion of results and no historical control data.

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Umwelt, Raumordnung und Landwirtschaft des Landes Nordrhein-Westfalen, Düsseldorf, Germany

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

853947-59-8

Cas Number:

853947-59-8

IUPAC Name:

853947-59-8

Method

Target gene:

Not applicable

Species / strain

Species / strain / cell type:

Chinese hamster lung fibroblasts (V79)

Details on mammalian cell type (if applicable):

- Type and identity of media:
MEM medium supplemented with 2 mM L-Glutamine and
- 4% (v/v) fetal calf serum (MEM4) or
- 0% (v/v) fetal calf serum (MEM0)
- 100 IU/mL penicillin/streptomycin
During exposure to the test substancewith S9 mix, MEM0 medium was used and replaced by MEM4 after 3 h after test substance administration.

Additional strain / cell type characteristics:

other: modal chromosome number of 22 and a cell cycle length of approx. 16.5 h

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

18h treatment: 10, 40 and 80 µg/mL (without metabolic activation)
18h treatment: 10, 60, 80 and 100 µg/mL (with metabolic activation)
28h treatment: 80 µg/mL (without metabolic activation)
28h treatment: 100 µg/mL (with metabolic activation)

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

MEM4 medium (resp. MEM0 medium in the test with S9 mix) containing 1% (v/v) ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: cyclophosphamide, 3 and 4 µg/mL in MEM0 medium, +S9; mitomycin C, 0.03 and 0.04 µg/mL in MEM4 medium, -S9

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 18 and 28 h
- Fixation time (start of exposure up to fixation or harvest of cells): 18 h treatment: 18 h; 28 h treatment: 28 h

STAIN (for cytogenetic assays): Giemsa

NUMBER OF REPLICATIONS: 2 replications each in one (28 h treatment) or two (18 h treatment) independent experiment

NUMBER OF CELLS EVALUATED: 100 per culture

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of 1000 cells per slide

OTHER EXAMINATIONS:
- Determination of polyploidy: yes
- Determination of endoreduplicated cells: yes

Evaluation criteria:

The test chemical is to be considered clastogenic in this assay if:
- it induces chromosomal aberrations (excl. gaps) in a statistically significant manner in one or more concentrations
- the induced proportion of aberrant cells at such test substance concentrations exceeds the normal range of the test system
- positive results can be verified in an independent experiment.

Statistics:

The number of aberrant cells in each replica was used to establish acceptable homogeneity between replicates by means of a binomial dispersion test ( Richardson, C., Williams, D. A., Allen, J. A., Amphlett, G., Chanter, D. O. and Phillips, B. Analysis of Data from In Vitro Cytogenetic Assays, in: Statistical Evaluation of Mutagenicity Test Data, Kirkland, D. J., (ed) Cambridge University Press, Cambridge, pp. 41-64., 1990 ). The proportion of cells that was treated with the test substance and harboured structural aberrations (excl. gaps) was compared with the corresponding proportion of the negative controls in the Chi-square test. Probability values of p < 0.05 were accepted as statistically significant.

Results and discussion

Test results

Species / strain:

Chinese hamster lung fibroblasts (V79)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

systematic influence of the test compound which led to a reduction in the mitotic index from 10 µg/mL.

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: The test substance was soluble in ethanol, in MEM4 medium containing 1% ethanol the solubility limit of the test substance was determined to be 100 µg/mL (homogeneous emulsion).

- Other confounding effects: In the first test without metabolic activation the negative controls exhibited only a mitotic index of 2.0%. Therefore, test 1 without metabolic activation was completely repeated with the same doses and named test #1a.

Any other information on results incl. tables

Table 1. Summary of data obtained in experiment #1a.

Test item	Concentration	Mitotic Index	Aberrant cells in %
	in µg/mL	in % mean	with gaps	without gaps
Exposure period 18h, without S9 mix
Control, Ethanol	1.0% (v/v)	10.7	2.0	0.0
MMC	0.03	2.1	24.6	18.5*
MMC	0.04	2.9	40.0	31.9*
Test substance	10	8.0	3.5	1.5
	40	6.7	3.0	0.5
	80	5.3	5.0	0.0
Exposure period 18h, with S9 mix
Control, Ethanol	1.0% (v/v)	7.0	6.5	4.0
CP	3.0	2.7	45.0	34.0*
Test substance	10	6.7	3.5	1.5
	60	6.1	4.5	1.0
	100	5.7	4.0	2.0

MMC: Mitomycin C; CP: Cyclophosphamide (positive controls) 

*: significant, no statistical evaluation

Table 2. Summary of data obtained in experiment #2.

Test item	Concentration	Mitotic Index	Aberrant cells in %
	in µg/mL	in % mean	with gaps	without gaps
Exposure period 18h, without S9 mix
Control, Ethanol	1.0% (v/v)	7.9	6.0	3.5
MMC	0.03	3.4	23.5	14.0*
Test substance	10	7.5	4.0	0.5
	40	8.0	9.5	3.0
	80	5.8	5.5	0.5
Exposure period 18h, with S9 mix
Control, Ethanol	1.0% (v/v)	6.3	8.0	2.0
CP	3.0	1.6	42.0	33.3*
Test substance	10	6.5	5.5	0.0
	60	5.7	4.5	1.5
	100	6.7	6.5	1.5
Exposure period 28h, without S9 mix
Control, Ethanol	1.0% (v/v)	7.9	3.5	0.0
MMC	0.03	4.4	43.5	32.5*
Test substance	80	5.8	4.5	0.5
Exposure period 28h, with S9 mix
Control, Ethanol	1.0% (v/v)	9.2	6.5	1.5
CP	3.0	6.0	36.5	27.5*
Test substance	100	8.4	3.5	1.0

MMC: Mitomycin C; CP: Cyclophosphamide (positive controls)

 *: significant, no statistical evaluation

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative