Cinnamyl alcohol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

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 journal.

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Gene mutation toxicity study of the test chemical

Author:

Sekizawa, J., Shibamoto, T.

Year:

1982

Bibliographic source:

Mutation Research, 1982

Reference 2

Reference Type:

publication

Title:

Gene mutation toxicity study of the test chemical

Author:

D. Bickers et. al.

Year:

2005

Bibliographic source:

Food and Chemical Toxicology (2005)

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

In vitro bacterial reverse gene mutation assay for test chemical was studied in S. typhimurium and Escherichia coli strains. The mutagenicity assay was conducted as described by Ames et al. with slight modifications. All tester strains were examined periodically for the markers indicated by Ames et al.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Cinnamyl alcohol

EC Number:

203-212-3

EC Name:

Cinnamyl alcohol

Cas Number:

104-54-1

Molecular formula:

C9H10O

IUPAC Name:

3-phenylpropan-1-ol

Test material form:

liquid

Remarks:

Oily liquid

Details on test material:

- Name of test material (as cited in study report): Cinnamyl alcohol
- IUPAC name : 3-Phenylprop-2-en-1-ol
- Molecular Formula: C9H10O
- Molecular Weight: 134.177 g/mol
- Smiles: c1(ccccc1)/C=C/CO
- InChI: 1S/C9H10O/c10-8-4-7-9-5-2-1-3-6-9/h1-7,10H,8H2/b7-4+
- Substance type: Organic
- Physical state: Crystalline solid

Method

Target gene:

Histidine for Salmonella strains and tryptophan for E. coli strains

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium, other: TA100, TA1535, TA98, TA1537, TA1538

Additional strain / cell type characteristics:

not specified

Species / strain 2

Species / strain / cell type:

E. coli WP2

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Type and composition of metabolic activation system: S9 mix (500 µM) consisted of 100µL of S9, 50 µmoles sodium phosphate buffer (pH 7.4), 4 µmoles MgCl2, 16.5 µmoles KCl, 2.5 µmoles G-6-P, 2 µmoles NADH and 2 µmoles NADPH.
- source of S9 : PCB-treated male Sprague-Dawley rats
- method of preparation of S9 mix : No data
- concentration or volume of S9 mix and S9 in the final culture medium : No data
- quality controls of S9 (e.g., enzymatic activity, sterility, metabolic capability): No data

Test concentrations with justification for top dose:

0, 250, 750, 1500 or 3000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: No data available

Controls

Controls 1

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

9-aminoacridine

2-nitrofluorene

sodium azide

Controls 2

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

benzo(a)pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION:
For Salmonella strains:
The assays without S9 were performed by the plate-incorporation method.
The assays with S9 were conducted by the pre-incubation method described by Yahagi et al. (1975). The pre-incubation method is useful in detecting weak mutagenicity in samples (Yahagi et al., 1977).. Histidine-independent colonies were scored after incubation at 37°C for 48-72 h.

For E. coli strains:
The assay was performed in the same manner as with the Salmonella assay except that the supplement of 0.1 µmole histidine plus 0.1 mole biotin in the soft agar was replaced with a supplement of 0.1 µmole of tryptophan. Tryptophan-independent revertant colonies were scored with E. coli.

DURATION
- Preincubation period: 37°C
- Exposure duration: 48-72 h.

Rationale for test conditions:

No data available

Evaluation criteria:

A bacterial cell line was observed for biologically relevant increase in the number of revertants. Revertants per plate represent average values from 3 to 5 replications.

Statistics:

No data available

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA100, TA1535, TA98, TA1537, TA1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

All test compounds showed killing on bacteria at the highest doses used.

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: RESULTS OF MUTAGENICITY TEST OF THE TEST CHEMICAL

Chemical	Dose (µg/plate)	Revertants/plate (mean±S.D.)
		Salmonella typhimuriumstrains					Escherichia coli
Without S9
DMSO	50µL	99±6	8±2	24±4	4±2	15±3	55±6
Positive control		769±54	266±34	660±30	822±88	301±33	452±68
Test chemical	250	98±5	10±4	28±1	3±1	9±1	50±9
	750	81±6	5±1	24±1	3±1	10±2	57±6
	1500	81±13	7±1	22±8	5±0	14±0	46±3
	3000	24±13	2±1	20±8	2±2	6±1	33±6
With S9
DMSO	50µL	106±9	10±3	32±4	7±2	21±4	59±5
Positive control		431±92	158±45	180±7	125±37	154±17	753±95
Test chemical	250	87±1	7±2	31±3	9±2	24±8	58±6
	750	130±3	10±1	27±3	7±2	18±4	62±3
	1500	123±12	9±2	24±3	8±3	21±3	42±4
	3000	72±11	12±2	21±2	4±2	11±0	39±8

Applicant's summary and conclusion

Conclusions:

The test chemical was negative in Ames test carried out using Salmonella typhimurium strains TA100, TA1535, TA98, TA1537, TA1538 as well in Escherichia coli WP2 uvr A in the presence and absence of S9 metabolic activation. Hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

In vitro bacterial reverse gene mutation assay for test chemical was studied using S. typhimurium and Escherichia coli WP2 uvrA strains. The mutagenicity assay was conducted as described by Ames et al. with slight modifications. Ames test was performed using Salmonella typhimurium strains TA100, TA1535, TA98, TA1537 and TA1538 and Escherichia coli WP2 uvrA strains in the presence and absence of S9 metabolic activation system at dose level of 0, 250, 750, 1500 or 3000 µg/plate. For Salmonella strains, the assays without S9 were performed by the plate-incorporation method and the assays with S9 were conducted by the pre-incubation method described by Yahagi et al. (1975). The pre-incubation method is useful in detecting weak mutagenicity in samples (Yahagi et al., 1977). Histidine-independent colonies were scored after incubation at 37°C for 48-72 h. For E. coli strains, the assay was performed in the same manner as with the Salmonella assay except that the supplement of 0.1 µmole histidine plus 0.1 mole biotin in the soft agar was replaced with a supplement of 0.1 µmole of tryptophan. Tryptophan-independent revertant colonies were scored with E. coli. No mutagenicity was noted at the tested dose levels. Based on the observations made, the test chemical was negative in Ames test carried out using Salmonella typhimurium strains TA100, TA1535, TA98, TA1537, TA1538 as well in Escherichia coli WP2 uvr A in the presence and absence of S9 metabolic activation. Hence it is not likely to classify as a gene mutant in vitro.