Hexa-2,4-dienoic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro DNA damage and/or repair study

Remarks:

Type of genotoxicity: DNA damage and/or repair

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1989

Report date:

1989

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.18 (DNA Damage and Repair - Unscheduled DNA Synthesis - Mammalian Cells In Vitro)

Deviations:

yes

Remarks:

Only 4.0E+05 cells were exposed to the test substance instead of at least 1.0E+06 cells.

GLP compliance:

no

Remarks:

The study was conducted in compliance with Good Laboratory Practice Regulations but prior to implementation of GLP.

Type of assay:

DNA damage and repair assay, unscheduled DNA synthesis in mammalian cells in vitro

Test material

Test material information

Constituent 1

Reference substance name:

Hexa-2,4-dienoic acid

EC Number:

203-768-7

EC Name:

Hexa-2,4-dienoic acid

Cas Number:

110-44-1

Molecular formula:

C6H8O2

IUPAC Name:

hexa-2,4-dienoic acid

Details on test material:

- Name of test material: Sorbic acid
- Physical state: solid, white powder
- Analytical purity: 99 %
- Stability under test conditions: The test substance was considered to be stable for the duration of the study.
- Storage condition of test material: 4 °C, protected from light

Method

Target gene:

not applicable

Species / strain

Species / strain / cell type:

mammalian cell line, other: Human cell line A 549 (American type culture collection no. CCL 185)

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

Experiment 1: 0, 1, 3, 10, 30, 100, 300, 1000 and 2000 µg/mL
Experiment 2: 0, 1, 3, 10, 30, 100, 300 and 1000 µg/mL

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: at the day of the expeiment the test substance was dissolved in DMSO (1 %)

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

without metabolic activation

Migrated to IUCLID6: 1 µg/mL

Controls 2

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

with metabolic activation

Migrated to IUCLID6: 10 µg/mL

Details on test system and experimental conditions:

METHOD OF APPLICATION: - two independent experiments with and without metabolic activation were performed

DURATION
- Exposure duration: 3 hours at 37 °C with or without 100 µg of S9 mix
NUMBER OF REPLICATIONS: 6

NUMBER OF CELLS EVALUATED: 4 x 10E+05

DETERMINATION OF CYTOTOXICITY: yes

OTHER EXAMINATIONS:
- The effect of the test substance on the frequency of mutation was tested by measuring incorporation of radioactively labelled thymidine into DNA determined by liquid scintillation counting (LSC).
- The DNA concentration was determined colourmetrically using diphenylamine reaction of deoxyribonucleic acid.

Evaluation criteria:

stat. significant dose related increase in incorporation (dpm/µg DNA)

Statistics:

Student's t-test

Results and discussion

Test results

Key result

Species / strain:

mammalian cell line, other: Human cell line A 549 (American type culture collection no. CCL 185)

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

with and without S9-mix

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At the highest concentration of 2000 µg/ml Sorbic acid, a visible microscopic alteration of the cell morphology was observed indicating cytotoxicity.

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'. Remarks: mammalian cells

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Sorbic acid did not cause any dose-related or reproducible increases in the mutant frequencies in mammalian cell line A 549 with or without metabolic activation. Statistically significant increases in gene mutation were observed for both positive control substances when compared to the negative controls (p <= 0.001). It was concluded that Sorbic acid is not genotoxic under the conditions of this test.

The extrapolation from sorbic acid to potassium sorbate or vice versa is considered not to be restricted in any way, since the determinant of potential toxicity is on the "sorbate" anion.