3-phenylpropionic acid

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:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

A Compilation of Two Decades of Mutagenicity Test Results with the Ames Salmonella typhimurium and L5178Y Mouse Lymphoma Cell Mutation Assays

Author:

H. E. Seifried, R. M. Seifried, J. J. Clarke, T. B. Junghans and R. H. C. San

Year:

2006

Bibliographic source:

Chem. Res. Toxicol. 2006, 19, 627-644

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

The gene mutation study was conducted according to L5178Y TK+/- Mouse Lymphoma Mutagenicity Assay to determine the mutagenic nature of 3-Phenylpropionic acid

GLP compliance:

not specified

Type of assay:

other: Mouse lymphoma assay

Test material

Test material information

Constituent 1

Reference substance name:

3-phenylpropionic acid

EC Number:

207-924-5

EC Name:

3-phenylpropionic acid

Cas Number:

501-52-0

Molecular formula:

C9H10O2

IUPAC Name:

3-phenylpropanoic acid

Details on test material:

SMILES:OC(=O)CCc1ccccc1

Specific details on test material used for the study:

- Name of test material: 3-Phenylpropionic acid
- IUPAC name: 3-phenylpropanoic acid
- Molecular formula: C9H10O2
- Molecular weight: 150.176 g/mol
- Substance type: Organic
- Physical state: No data available.
- Purity: No data available
- Impurities (identity and concentrations): No data available

Method

Target gene:

Thymidine kinase

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Remarks:

TK+/- 3.7.C

Details on mammalian cell type (if applicable):

- Type and identity of media:
The cells were grown in Fischer’s medium for leukemic cells of mice supplemented with 10% horse serum and 0.02% pluronic F-68.
- Properly maintained: No data available
- Periodically checked for Mycoplasma contamination: Yes
- Periodically checked for karyotype stability: No data available
- Periodically "cleansed" against high spontaneous background: No data available

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

liver S9 prepared from Aroclor 1254-induced male Sprague- Dawley rats.

Test concentrations with justification for top dose:

500 - 8091 µg/mL

Vehicle / solvent:

No data available

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

other: ethyl methylsulfonate, 3-methylcholanthrene and dimethylbenz[a]- anthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: No data available
- Exposure duration: 4 h
- Expression time (cells in growth medium):48 h
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): 1×106 cells/plate for mutant selection
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: Duplicate

NUMBER OF CELLS EVALUATED: 1X 106 cells/plate for mutant selection and 200
cells/plate for viable count determinations

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

OTHER: No data available

Rationale for test conditions:

No data

Evaluation criteria:

Results were interpreted using a doubling of the mutant frequency over the concurrent solvent-treated control value as an indication of a positive effect, together with evidence of a dose-related increase. Doubling of the mutant frequency was previously reported as representing a positive effect. Only doses yielding total growth values of 10% were used in the analysis of induced mutant frequency. Doses yielding less than 10% total growth were used in determining dose response.

Statistics:

No data

Results and discussion

Test results

Species / strain:

mouse lymphoma L5178Y cells

Remarks:

TK+/- 3.7.C

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: No data available
- Effects of osmolality: No data available
- Evaporation from medium: No data available
- Water solubility: No data available
- Precipitation: No data available
- Other confounding effects: No data available

RANGE-FINDING/SCREENING STUDIES: No data available

COMPARISON WITH HISTORICAL CONTROL DATA: No data available

ADDITIONAL INFORMATION ON CYTOTOXICITY: The doses of chemical selected for testing were within the range yielding approximately 0-90% cytotoxicity.

Applicant's summary and conclusion

Conclusions:

3-Phenylpropionic acid failed to induce a doubling of the mutant frequency both in the presence and absence of S9 activation system and hence is not likely to be gene mutant in vitro.

Executive summary:

The gene mutation study was conducted according toL5178Y TK+/-Mouse Lymphoma Mutagenicity Assay to determine the mutagenic nature of the test compound 3-Phenylpropionic acid.

The Cells at a concentration of 1.2 X 10⁷cells/mL were exposed for 4 h to a range of concentrations from 500 -8091 µg/mL. The cells were then washed, resuspended in growth medium, and incubated at 37°C for 48h to allow recovery and mutant expression. Cells in the cultures were adjusted to 3 X 10⁵/mL at 24 h intervals. They were then cloned (1 X 10⁶cells/plate for mutant selection and 200 cells/plate for viable count determinations) in soft agar medium containing Fischer’s medium, 20% horse serum, 2 mM sodium pyruvate, 0.02% pluronic F-68, and 0.23% granulated agar. Resistance to trifluorothymidine (TFT) was determined by adding TFT (final concentration, 3µg/mL) to the cloning medium for mutant selection. The 100X stock solution of TFT in saline was stored at -70 °C and was thawed immediately before use. Plates were incubated at 37 ( 1 °C in 5% CO2 in air for 10-12 days and then counted with an Artek automated colony Counter. Only colonies larger than ~ 0.2 mm in diameter were counted. Mutant frequencies were expressed as mutants per 10⁶surviving cells.

3-Phenylpropionic acid failed to induce a doubling of the mutant frequency both in the presence and absence of S9 activation system and hence is not likely to be gene mutant in vitro.