p-methoxybenzyl acetate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000-07-16 and 2000-07-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

p-methoxybenzyl acetate

EC Number:

203-185-8

EC Name:

p-methoxybenzyl acetate

Cas Number:

104-21-2

Molecular formula:

C10H12O3

IUPAC Name:

4-methoxybenzyl acetate

Method

Target gene:

Salmonella typhimurium: histidine

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

Liver homogenate (S9) from Aroclor 1254 pretreated male rats

Test concentrations with justification for top dose:

0, 15, 50, 150, 500, 1500, 5000 µg/plate

Vehicle / solvent:

- Vehicle/solvent used: DMSO

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (TA 98, TA 100, TA 102, TA 1535, TA 1537)

Remarks:

with metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 to 72 h

NUMBER OF REPLICATIONS: two independent experiments with three replications

DETERMINATION OF CYTOTOXICITY
- Method: Number of revertant colonies

Evaluation criteria:

The test substance is evaluated as mutagenic if it significantly or dose dependently increases the mutation frequency of the tester strains.

Statistics:

For estimation of the statistical significance of the difference between the mean number of revertants in the negative controls and the plates at each dosage level a X2-test was used.

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA1537, TA1535, TA98, TA100 and TA102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Bacteriotoxic towards the strains TA 1537 and TA 102 at 5000 µg/plate without S9 mix.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation was observed.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Bacteriotoxic towards the strains TA1537 and TA102 at 5000μg/plate in the absence of S9-mix.

Any other information on results incl. tables

 Experiment 1:

Dose (µg/plate)	Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium
	TA98	TA100	TA102	TA1535	TA1537
	Results without S9
Untreated	30± 5	122 ± 9	279 ± 25	15 ± 1	8 ± 2
DMSO (50µL)	25 ± 5	134 ± 5	246 ± 12	15 ± 3	9 ± 6
15			252 ± 13
50	24 ± 6	123 ± 6	243 ± 16	15 ± 4	7 ± 3
150	19 ± 2	123 ± 6	241 ± 16	15 ± 3	5 ± 3
500	30 ± 6	139 ± 1	215 ± 11	17 ± 3	9 ± 4
1500	15 ± 6	134 ± 12	197 ± 19	12 ± 4	8 ± 3
5000	21 ± 6	147 ± 5	108 ± 5	13 ± 4	4 ± 3
NaN3 (0.7)		483 ± 15		428 ± 20
2-NF (2.5)	368 ± 24
9-AA (50)					187 ± 13
Mitomycin (0.15)			727 ± 7
	Results with S9
Untreated	26 ± 7	149 ± 6	267 ± 10	9 ± 1	17 ± 6
DMSO (50µL)	21 ± 4	131 ± 8	147 ± 4	10 ± 2	16 ± 2
15			243 ± 8		10 ± 5
50	21 ± 6	133 ± 3	247 ± 12	12 ± 3	17 ± 4
150	35 ± 5	132 ± 9	251 ± 10	14 ± 2	11 ± 3
500	19 ± 3	131 ± 2	235 ± 10	13 ± 4	9 ± 5
1500	29 ± 4	133 ± 6	249 ± 14	10 ± 3	14 ± 4
5000	19 ± 2	140 ± 8	230 ± 2	9 ± 3
2-AA (0.7)	1081 ± 29
2-AA (0.8)		2394 ± 162
2-AA (1.0)				523 ± 29
2-AA (1.5)			1253 ± 45		210 ± 17

2-NF: 2-nitrofluorene; 9-AA: 9-aminoacridine; 2-AA: 2-aminoanthracene

 

 

Experiment 2:

Dose (µg/plate)	Mean number of revertant colonies/3 replicate plates (± S.D.) with different strains of Salmonella typhimurium
	TA98	TA100	TA102	TA1535	TA1537
	Results without S9
Untreated	16 ± 2	115 ± 11	249 ± 6	16 ± 3	8 ± 4
DMSO (50µL)	14 ± 5	123 ± 4	237 ± 11	14 ± 3	9 ± 4
15			223 ± 9
50	14 ± 5	106 ± 9	240 ± 16	12 ± 4	9 ± 4
150	17 ± 2	116 ± 7	234 ± 9	12 ± 2	9 ± 4
500	13 ± 2	112 ± 11	198 ± 11	18 ± 4	6 ± 3
1500	14 ± 1	120 ± 10	195 ± 2	19 ± 4	9 ± 2
5000	10 ± 4	134 ± 4	111 ± 12	14 ± 3	5 ± 6
NaN3 (0.7)		334 ± 24		180 ± 15
2-NF (2.5)	339 ± 41
9-AA (50)					331 ± 38
Mitomycin (0.15)			1106 ± 112
	Results with S9
Untreated	25 ± 6	129 ± 11	255 ± 6	12 ± 3	19 ± 5
DMSO (50µL)	21 ± 2	120 ± 5	245 ± 8	10 ± 3	14 ± 6
50	20 ± 0	109 ± 1	223 ± 13	8 ± 2	13 ± 7
150	20 ± 3	109 ± 8	245 ± 7	13 ± 2	11 ± 2
500	24 ± 5	114 ± 3	230 ± 15	11 ± 5	11 ± 4
1500	21 ± 7	125 ± 9	204 ± 3	11 ± 3	13 ± 4
5000	21 ± 3	113 ± 13	193 ± 9	6 ± 2	14 ± 4
2-AA (0.7)	1207 ± 57	1665 ± 51
2-AA (1.0)				313 ± 8
2-AA (1.5)			1265 ± 25		282 ± 23

2-NF: 2-nitrofluorene; 9-AA: 9-aminoacridine; 2-AA: 2-aminoanthracene

Applicant's summary and conclusion

Conclusions:

The test substance was not mutagenic to Salmonella typhimurium strains TA1535, TA1537, TA98, TA100, and TA102 in the presence and absence of a metabolizing system.

Executive summary:

An in vitro reverse mutation assay study (Ames) was conducted according to OECD Guideline 471 in Salmonella typhimurium (TA98, TA100, TA102, TA1535 and TA1537). Experiments were performed as a plate incorporation assay. All tests were conducted in triplicate and concentrations between 15 μg/plate and 5000 μg/plate were tested. Strains were exposed to the test substance with and without metabolic activation with rat liver S9 mix. Cytotoxicity was observed without S9 in the strains TA 1537 and TA 102 at 5000 μg/plate. No substantial increase in revertant colony numbers of any of the tester strains was detected following treatment with the test substance at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance. Therefore it was concluded that the test substance did not induce gene mutations and it was considered to be non-mutagenic.