Fatty acids, C16-18 and C18-unsatd., tetraesters with pentaerythritol

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:

06. Oct. - 16. Feb. 2004

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD). (Purity of test substance not given, evalation criteria not given.)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

yes

Remarks:

purity of test substance is not given (responsibility of the sponsor)

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Test material

Test material information

Constituent 1

Reference substance name:

403507-18-6

EC Number:

609-825-6

Cas Number:

403507-18-6

IUPAC Name:

403507-18-6

Details on test material:

- Name of test material (as cited in study report): only trade name given
- Lot/batch No.: AO-267-108
- Storage condition of test material: room temperature in the dark

Method

Target gene:

not applicable

Species / strain

Species / strain / cell type:

lymphocytes: cultured peripheral human lymphocytes

Details on mammalian cell type (if applicable):

- Type and identity of media: Eagles essential medium with HEPES buffer (MEM), supplemented with:
L-glutamine, penicillin/streptomycin, amphotericin B, 15% foetel calf serum
- Properly maintained: yes

Metabolic activation:

with and without

Metabolic activation system:

co-factor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats pretreated with phenobarbitone (80 mg/kg) and ß-naphtoflavone (100 mg/kg)

Test concentrations with justification for top dose:

Experiment I:
4 hour (with and without): 40, 80, 160, 240*, 320*, 400* µg/mL

Experiment II
4 hour (with): 40, 80, 160, 240*, 320*, 400* µg/mL
24 hour (without): 40, 80, 160, 240*, 320*, 400* µg/mL

* Dose levels (plus control dose) selected for metaphase analysis

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Mitomycin C (MMC; 0.2 and 0.4 µg/mL; -S9), cyclophosphamide (CP; 10 µg/mL; +S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 h (with and without S9), 24 h (without)
- Fixation time (start of exposure up to fixation or harvest of cells): 4 h treatment: 20 h; 24 h treatment: 0 h

SPINDLE INHIBITOR (cytogenetic assays): Colcemid 0.1 µg/mL (demecolcine)
STAIN (for cytogenetic assays): 5% Gurrs Giemsa for 5 minutes

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: 200 per culture

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index of 2000 cells

OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

no data

Statistics:

The frequency of cells with aberrations excluding gaps and the frequency of polyploid cells was compared, where necessary, with the concurrent vehicle control value using Fisher's exact test.

Results and discussion

Test results

Species / strain:

lymphocytes: cultured peripheral human lymphocytes

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: cloudy precipitates were observed at and above 40 and 80 µg/mL in the 24-hour continuous and 4-hour pulse treatment groups, respectively

RANGE-FINDING/SCREENING STUDIES:
The dose range tested was 10-320 µg/mL. The test material produced some weak toxicity in the 4-hour treatment group but not the 24-hour treatment group. Toxicity could not be reproduced in the main experiment (scorable metaphases at every dose level).
COMPARISON WITH HISTORICAL CONTROL DATA:
The results are in range with historical control data.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 3 + 4: Test results of experiment I.

Test item	Concentration	Mitotic Index	Aberrant cells in %
Experiment I	in µg/mL	in %	with gaps	without gaps
Exposure period 4h, fixation time 20h, without S9 mix
control	0	100	1	0
MMC	0.4	35	53	37
Test substance	240	98P	0.5	0
	320	110P	0	0
	400	91P	0.5	0.5
Exposure period 4h, fixation time 20h, with S9 mix
control	0	100	1	0.5
CP	10	20	35.5	28.5
Test substance	240	89P	1.5	0
	320	89P	0.5	0
	400	108P	3.5	1

Test item	Concentration	Mitotic Index	Aberrant cells in %
Experiment II	in µg/mL	in %	with gaps	without gaps
Exposure period + fixation time 24h, without S9 mix
control	0	100	1.5	0
MMC	0.4	41	70	67
Test substance	240	61	0.5	0.5
	320	53	0.5	0
	400	83	0	0
Exposure period 4h, fixation time 20h, with S9 mix
control	0	100	0.5	0
CP	10	30	67	56
Test substance	240	103	1	0
	320	76	1	0
	400	110	1.5	0.5

Table5 +6: Mean Frequency of Polyploid Cells (%)

Experiment I dose level µg/mL	harvest time 24 hours
	4 hours without S9	4 hours with S9
0	0.0	0.0
240	0.0	0.0
320	0.0	0.0
400	0.0	0.0
MMC 0.4	0.0	NA
CP 10	NA	0.0

dose level µg/mL	harvest time 24 hours
	24 hours without S9	4 hours with S9
0	0.5	0.0
240	0.0	0.0
320	0.0	0.0
400	0.0	0.5
MMC 0.4	0.0	NA
CP 10	NA	0.0

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative