Diisononyl adipate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

publication

Title:

An Evaluation of the Genotoxic Potential of Di-lsononyl Adipate

Author:

McKee

Year:

1986

Bibliographic source:

Environ Mutagen 8:817-827

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

no

Type of assay:

mammalian cell gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Diisononyl adipate

EC Number:

251-646-7

EC Name:

Diisononyl adipate

Cas Number:

33703-08-1

Molecular formula:

C24H46O4

IUPAC Name:

1,6-bis(7-methyloctyl) hexanedioate

Test material form:

liquid

Specific details on test material used for the study:

- Name of test material (as cited in study report): DINA, Cas-No. 33703-08-1
- Analytical purity: 99 %

Method

Target gene:

thymidine kinase

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced S9-mix

Test concentrations with justification for top dose:

without S9-mix: 0, 7.5, 10, 13, 18, 24, 56, 75, 100 µl/mL
with S9-mix: 0, 5.6, 10, 13, 18, 24, 32, 42, 56, 75, 100 µl/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: due to solubility

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

ethylmethanesulphonate

Details on test system and experimental conditions:

The mouse lymphoma gene mutation assay was conducted essentially as described by Clive and co-workers [1973, 1979; Clive and Spector, 1975]. Suspension cultures of mouse lymphoma cells, heterozygous for thymidine kinase activity, were grown in Fisher medium for leukemic mouse cells supplemented with 0.1% pluronic and 10% heat-inactivated horse serum (Fl0P) and were exposed to test substances in the same medium. Treated cells were grown in Fl0P for 48 hr to allow mutation expression. Approximately 3 x l0^6 cells from each culture were then plated in a selective medium containing 3 µg/ml trifluorothymidine (TFT) to select for mutant clones. Appropriately diluted cells from each culture were also seeded in plates without TFT for viability determinations (200 cells/plate). Mutant and total colony counts at each dose point were determined by triplicate plating. Colonies were counted with an automatic colony counter; differential sizing was not performed.

Evaluation criteria:

The validity of the experimental assay was verified through the use of concurrent positive, control materials, 7,12dimethyl-benzanthracene (DMBA) and ethylmethane sulfonate (EMS). Increased mutation frequencies induced by the positive control materials under appropriate conditions demonstrated both the integrity of the metabolic activation system and the responsiveness of the assay system as compared to its historical performance in the laboratory.

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: no data

COMPARISON WITH HISTORICAL CONTROL DATA: yes

ADDITIONAL INFORMATION ON CYTOTOXICITY:

Applicant's summary and conclusion

Conclusions:

The provided mammalian cell mutation assay in mouse lymphoma cells provided an overall negative result for the testes substance.

Executive summary:

In an mouse lymphoma test (similar to OECD 476) DINA was tested with and without S9-mix in a dose range of 5.6 - 1000 µl/ml. The test substance was dissolved in acetone. There was no evidence of a mammalian cell mutation in the mouse lymphoma assay (McKee, 1985)