Benzyl phenylacetate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

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 CEBS (Chemical Effect in Biological System) report

Data source

Reference

Reference Type:

publication

Title:

Gene mutation toxicity study for Benzyl phenylacetate

Author:

National Institute of Environmental Health Sciences (NIEHS)

Year:

2015

Bibliographic source:

Chemical Effect in Biological System, CEBS no. 002-01716-0001-0000-8

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:

Gene mutation study by the preincubation assay was performed to determine the mutagenic nature of benzyl phenylacetate

GLP compliance:

no

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Benzyl phenylacetate

EC Number:

203-008-4

EC Name:

Benzyl phenylacetate

Cas Number:

102-16-9

Molecular formula:

C15H14O2

IUPAC Name:

benzyl phenylacetate

Details on test material:

Name of test material (as cited in study report): Benzyl phenylacetate
Substance type: Organic
Physical state: Liquid

Specific details on test material used for the study:

- Name of test material: Benzyl phenylacetate
- Molecular formula: C15H14O2
- Molecular weight: 226.2736 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: No data
- Impurities (identity and concentrations): No data

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA97, TA98, TA100 and TA1535

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

10% and 30% rat liver and hamster liver S9 fraction

Test concentrations with justification for top dose:

TA100 and TA98: 0, 0.3, 1, 3, 10, 33, 66, 100, 333, 1000, 3333 or 10000 µg/plate
TA 1535 and TA1537: 0, 0.3, 1, 3, 10, 33, 100, 333, 1000, 3333 or 10000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was solubel in DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION : Pre incubation

DURATION
No data

SELECTION AGENT (mutation assays):No data
SPINDLE INHIBITOR (cytogenetic assays):No data
STAIN (for cytogenetic assays):No data

NUMBER OF REPLICATIONS: No data

NUMBER OF CELLS EVALUATED:
No data

METHOD OF APPLICATION: preincubation
- Cell density at seeding (if applicable): No data

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

SELECTION AGENT (mutation assays): No data

SPINDLE INHIBITOR (cytogenetic assays): v

STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: No data

METHODS OF SLIDE PREPARATION AND STAINING TECHNIQUE USED: No data

NUMBER OF CELLS EVALUATED: No data

NUMBER OF METAPHASE SPREADS ANALYSED PER DOSE (if in vitro cytogenicity study in mammalian cells): No data

CRITERIA FOR MICRONUCLEUS IDENTIFICATION: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data
- Any supplementary information relevant to cytotoxicity: Yes, cytotoxicity was noted

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Methods, such as kinetochore antibody binding, to characterize whether micronuclei contain whole or fragmented chromosomes (if applicable): No data

- OTHER:

Rationale for test conditions:

No data

Evaluation criteria:

The plates were observed for a dose dependent increase in the number of revertants/plate

Statistics:

Mean ± standard error of mean

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA97, TA98, TA100 and TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Remarks:

Slight toxicity was noted

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No data

Any other information on results incl. tables

Strain: TA100

Dose	No Activation (Negative)		No Activation (Negative)		30% RLI (Negative)		30% HLI (Negative)		10% RLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	141	7.3	106	3.7	161	5.1	165	4.1	145	7	117	8.2
0.3			101	2.3
1	154	4.3	104	4
3	154	10.8	97	6.9
10	150	14.6	109	8
33	152	835	89	1.7
66	58 s	4.8
100					157	1.5	190	11.7	130	9.9	123	3.6
333					180	7.1	161	12.8	128	3.5	130	10.1
1000					163	1.7	190	9.7	122	5.8	118	10.1
3333					173	4.7	148	20.2	106	12.4	116	3.5
10000					153	7.9	186	17.3	104	13.8	105	5.9
Positive Control	466	15.3	7.4	58.5	443	42.6	638	10.9	422	16.5	587	43.3

 

Strain :TA1535

Dose	No Activation (Negative)		No Activation (Negative)		30% RLI (Negative)		30% HLI (Negative)		10% RLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	12 12	1.9	12	1.5	12	0.3	13	2.1	13	2	14	2.9
0.3	12	1.2	8	0.9
1	9	0.9	9	0.7
3	10	1.2	7	0.9
10	10	2.1	10	2.3
33	9	1	10	1.9
100					19.3	0.3	12	0.9	13	2.3	12	0.3
333					15	2.3	8	1.5	13	3.7	10	2.6
1000					12	0.3	4	1.2	8	2.2	12	1.7
3333					9	0.6	8	1.5	10	1.5	9	1.5
10000					8	1.2	7	0.7	8	2	10	2.2
Positive Control	585	19.9	635	37.5	106	10.8	256	24.9	71	8.1	61	0.9

 

Strain :TA97

Dose	No Activation (Negative)		No Activation (Negative)		30% RLI (Negative)		30% HLI (Negative)		10% RLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	203	6.2	199	0.9	231	8	218	10.9	188	5.5	168	10
0.3	207	4.1	179	3.7
1	223	8.5	125	5
3	199	7.5	182	10.4
10	189	4	176	7.2
33	191	4.8	185	13.7
100					237	11.7	227	2.6	190	5.6	187	8.6
333					242 c	15.5	211	5.9	220	9.7	189	15.8
1000					243	6.6	199	11.6	187	1.8	164	12.7
3333					254	15.7	204	16.3	148	10.4	146	4.3
10000					190	2.3	185	14.6	162	13.3	128	5
Positive Control	554	8.3	470	27.7	378	19.7	426	12.7	282	8.6	342	48.5

 

Strain :TA98

Dose	No Activation (Negative)		No Activation (Negative)		30% RLI (Negative)		30% HLI (Negative)		10% RLI (Negative)		10% HLI (Negative)		10% RLI (Negative)		10% HLI (Negative)
Protocol	Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation		Preincubation
µg/plate	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM	Mean	±SEM
0	22	3.1	21	3.7	23	3.1	20	2.6	27	3.2	20	2.4	23	0.3	22	5.8
0.3			17	0
1	24	2.5	20	4.4
3	27	1.8	12	3.2
10	22	1.7	19	2.8							21	2.3			16	2.1
33	15	1.5	17	0.6							19	1.5			15	1.5
66	4s	1.7
100					23	44	22	2.6	18	1.5	14	1.2	17	0.6	18	1
333					24	2.7	24	4.8	19	4.3	8	1.9	16	0.6	12	2.3
100					16 2	2	22	4.3	13	2.2	5s	0.9	11	1.2	11	0.9
3333					15	2.9	18	1.5	9s	2.3			7s	0.9
1000					12 s	1.9	10	2.2	5s	0.9			4s	1
Positive Control	399	23.8	894	16.8	129	3.8	345	9.1	332	30.3	341	15	473	18.5	584	7.8

RLI = induced male Sprague Dawley rat liver S9

HLI = induced male Syrian hamster liver S9

s = Slight Toxicity; p = Precipitate; x = Slight Toxicity and Precipitate; t = Toxic; c = Contamination

Applicant's summary and conclusion

Conclusions:

Benzyl phenylacetate did not induce a a dose dependent increase in the number of revertants per plate in Salmonella typhimurium strain TA97, TA98, TA1535 and TA100 with and without 10% and 30% rat liver and hamster liver S9 metabolic activation system and hence exhibited no mutagenic activity under the given test conditions.

Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of benzyl phenylacetate using preincubation assay. The study was performed using Salmonella typhimurium strain TA97, TA98, TA1535 and TA100 with and without 10% and 30% rat liver and hamster liver S9 metabolic activation system. The study was performed at dose level of 0, 0.3, 1, 3, 10, 33, 66, 100, 333, 1000, 3333 or 10000 µg/plate for TA100 and TA98 and at a dose level of

0, 0.3, 1, 3, 10, 33, 100, 333, 1000, 3333 or 10000 µg/plate for TA1535 and TA97 with DMSO as the solvent. The plates were observed for a dose dependent increase in the number of revertants/plate. Benzyl phenylacetate did not induce a dose dependent increase in the number of revertants per plate and hence exhibited no mutagenic activity under the given test conditions.