Bis(2-ethylhexyl) sebacate

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

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.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Reference 2

Reference Type:

publication

Title:

In Vitro Chromosomal Aberration Test of Bis(2-ethylhexyl) azelate in Cultured Chinese Hamster Cells

Author:

Miwa, Y.

Year:

2004

Bibliographic source:

Toxicity Testing Reports of Environmental Chemicals, 11, 287-320

Reference 3

Reference Type:

secondary source

Title:

Bis(2-ethylhexyl) azelate

Author:

OECD

Year:

2006

Bibliographic source:

SIDS Initial Assessment Report for SIAM 22, Paris, France, 18-21 April, 2006

Reference 4

Reference Type:

secondary source

Title:

Diesters Category of the Aliphatic Esters Chemicals (Test Plan and Robust Summaries for Substances in the HPV Test Plan)

Author:

US-EPA (American Chemistry Council's Aliphatic Esters Panel)

Year:

2010

Bibliographic source:

High Production Volume (HPV) Chemical Challenge Program (201-16837A and 201-16837B)

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:

103-24-2 (analytical purity 77.2%)

IUPAC Name:

103-24-2 (analytical purity 77.2%)

Details on test material:

- Name of test material (as cited in study report): Bis(2-ethylhexyl)nonanedioate
- Analytical purity: 77.2%
- Impurities (identity and concentrations):
Bis(2-ethylhexyl)glutarate 2.2%,
Bis(2-ethylhexyl)adipate 2.4%,
Bis(2-ethylhexyl)pimelate 2.8%,
Bis(2-ethylhexyl)suberate 3.8%,
Bis(2-ethylhexyl)sebacate 3.3%,
Bis(2-ethylhexyl) 1-,9-nonamethylenedicarboxylate 5.3%,
Bis(2-ethylhexyl)1-,10-decamethylenedicarboxylate 0.6%,
Bis(2-ethylhexyl)1-,11-undecamethylenedicarboxylate 0.3%
- Lot/batch No.: N-31101

Method

Target gene:

Not applicable

Species / strain

Species / strain / cell type:

mammalian cell line, other: Chinese hamster lung (CHL/IU) cell

Details on mammalian cell type (if applicable):

- Type and identity of media: Eagles's MEM modified with NaHCO3 and HCl (pH 7.0 - 7.1). Fetal calf serum was added up to 10%.
- Properly maintained: yes
- Periodically "cleansed" against high spontaneous background: yes

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phenobarbital and 5,6-benzoflavone.

Test concentrations with justification for top dose:

6 h treatment: 150, 300, 600, 1200, 2400 µg/mL with and without metabolic activation.
24 h treatment: 37.5, 75, 150, 300, 600 µg/mL without metabolic activation.

Vehicle / solvent:

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

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: Mitomycin C (MMC, 0.1 µg/mL ); +S9: Dimethylnitrosamin (DMN; 500 µg/mL)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 6 h and 24 h

NUMBER OF REPLICATIONS: duplicates in single experiment

NUMBER OF CELLS EVALUATED: 200

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index

OTHER EXAMINATIONS:
- Determination of polyploidy: yes

Evaluation criteria:

Evaluation was made on the basis of incidence as; -: negative (less than 5.0%); ±: equivocal (5.0% or higher to less than 10.0%); +: positive (10.0% or higher)

Results and discussion

Test results

Species / strain:

mammalian cell line, other: Chinese hamster lung (CHL/IU) cell

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: There were colorless clear fine oil drop-like precipitations and oil membrane-like precipitations in the culture fluid in petri plate at the concentrations of 1200 µg/mL or more.

RANGE-FINDING: The maximum concentration was established, based on the growth inhibition test. In this test, 50% growth inhibition was observed at 1859.0 µg/mL with S9 and at 1555.5 µg/mL without S9 for 6 h short treatment and 534.2 µg/mL without S9 for 24 h continuous treatment.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Test results

Test item	Concentration	Mitotic Index	Aberrant cells in %
	in µg/mL	in %	with gaps	without gaps
Exposure period 6 h, without S9 mix
Medium		99	0.5	0.5
Ethanol		100	1	1
MMC	0.1	83	55.5	55.5
Test substance	150	99	0	0
	300	88	0	0
	600	70	0.5	0.5
	1200 P	51	1	1
	2400 P	39	0.5	0.5
Exposure period 6 h, fixation time 24 h, with S9 mix
Medium		100	0	0
Ethanol		100	0	0
DMN	500	78	65.5	65.5
Test substance	150	99	1	1
	300	80	0	0
	600	64	0.5	0.5
	1200 P	48	0	0
	2400 P	35	0	0
Exposure period 24 h, fixation time 24 h, without S9 mix
Medium		97	0.5	0.5
Ethanol		100	1	1
MMC	0.05	86	44.5	44.5
Test substance	37.5	97	1.5	1.5
	75	98	0	0
	150	87	0	0
	300	56	1	1
	600	35	0	0

MMC: Mitomycin C

DMN: Dimethylnitrosamine

P: Colorless clear fine oile drop-like precipitations were noted in the culture fluid in petri plate.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative