2-propenoic acid, 2-methyl-, 4-benzoylphenyl ester

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:

22.10.2009 to 15.12.2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

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:

2-propenoic acid, 2-methyl-, 4-benzoylphenyl ester

EC Number:

611-390-2

Cas Number:

56467-43-7

Molecular formula:

C17H14O3

IUPAC Name:

2-propenoic acid, 2-methyl-, 4-benzoylphenyl ester

impurity 1

Reference substance name:

(4-benzoylphenyl) acetate

Cas Number:

13031-44-2

Molecular formula:

C15H12O3

IUPAC Name:

(4-benzoylphenyl) acetate

Test material form:

solid

Specific details on test material used for the study:

4-(Methacryloyloxy) bezophenone 97.26 %
Benzophenone acetate: 1.95 %

Method

Target gene:

his-locus
TA98: his D 3052; rfa"; uvrB·; R-factor: frame shift mutations
TA 100: his G 46; rfa·; uvrB"; R-factor: base-pair substitutions
TA 1535: his G 46; rfa·; uvrB": base-pair substitutions
TA 1537: his C 3076; rfa"; uvrB·: frame shift mutations
TA 102: his G 428 (pAQI ); rfa"; R-factor: base-pair substitutions

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:

S9 mix

Test concentrations with justification for top dose:

Experiment I:
10.0, 31.6, 100, 316, 1000, 2500 and 5000 μg/plate
3.16, 10.0, 31.6, 100, 316, 1000 and 2500 μg/plate (only TA I 02 without metabolic activation)
Experiment II:
3.16, 10.0,31.6, 100,316, 1000 and 2500 μg/plate (TA 98 and TA 1535 with and without metabolic activation)
1.00, 3.16, 10.0, 31.6, 100, 316 and 1000 μg/plate (TA 100 and TA 1537 with and without metabolic activation, TA 102 with metabolic activation)
0.316, 1.00, 3 .16, 10.0, 31.6, 100 and 316 μg/plate (TA 102 without metabolic activation)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility properties + relative nontoxicity to the bacteria

Controls

Controls 1

Positive controls:

yes

Remarks:

without metablic activation

Positive control substance:

sodium azide

Remarks:

Tester Strains: S. typhimurium: TA 100, TA 1535 Name: Sodium azide, NaN3 Purity: at least 99% Dissolved in: Aqua dest. Concentration: 10 µg/plate

Controls 2

Positive controls:

yes

Remarks:

without metabolic activation

Positive control substance:

other: 4-nitro-o-phenylene-diamine, 4-NOPD

Remarks:

Tester Strains: S. typhimurium: TA 98, TA 1537 Name: 4-nitro-o-phenylene-diamine, 4-NOPD Purity: >97% Dissolved in: DMSO Concentrations: 10 µg/plate forTA 98, 40 µ/plate for TA 1537

Controls 3

Positive controls:

yes

Remarks:

without metabolic activation

Positive control substance:

methylmethanesulfonate

Remarks:

Tester Strain: S. typhimurium: TA 102 Name: Methyl methane sulfonate, MMS Purity: 99.0% Dissolved in: Aqua dest. Concentration: 1 µg/plate

Controls 4

Positive controls:

yes

Remarks:

with metabolic activation

Positive control substance:

other: 2-aminoanthracene, 2-AA

Remarks:

Tester Strains: S. typhimurium: TA 98, TA 100, TA 1535, TA 1537 and TA 102 Purity: 96% Dissolved in: DMSO Concentrations: 2.5 µg/plate; 10 µg/plate for TA 102

Controls 5

Negative solvent / vehicle controls:

yes

Remarks:

Aqua dest.

Remarks:

Solvent controls, consisting of solvent or vehicle alone as well as untreated controls were treated in the same way as the treatment groups.

Controls 6

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Remarks:

Solvent controls, consisting of solvent or vehicle alone as well as untreated controls were treated in the same way as the treatment groups.

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation) and preincubation

DURATION
- Preincubation period: 60 minutes
- Exposure duration: at least 48 h

NUMBER OF EXPERIMENTS: 2 (one performed as pre-incubation assay, one as plate incorporation assay), 3 plates per concentration each

DETERMINATION OF CYTOTOXICITY
- Method: Toxicity of the test article may be evidenced by a reduction in the number of spontaneous revertants, a clearing of the bacterial background lawn, or by degree of survival of treated cultures.

OTHER:
automatic colony count with “AUTOCOUNT” (Artek systems, Biosys)

Evaluation criteria:

- corresponding background growth on negative control and test plates
- normal range of spontaneous revertants: TA1535 (10-29), TA 1537 (5-28), TA 1538 (12-37), TA 98 (15-57), TA 100 (77-189)
- test material is considered mutagenic if either a dose related and reproducible increase in revertant numbers or a significant and reproducible increase in revertant numbers for at least one test concentration in induced
- a significant increase in revertants means at least twice as high in TA 100 and a three times higher number in TA 1535, TA 1537, TA 1538, TA 89 compared to the negative control

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA98, TA 100, TA 1535, TA 1537, TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, 4-(Methacryloyloxy) benzophenone did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used.
Therefore, 4-(Methacryloyloxy) benzophenone is considered to be non-mutagenic in this bacterial reverse mutation assay.

Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471, adopted 21 July 1997, strains TA 1535, TA 1537, TA 98, TA 100 and TA 102 were exposed to 4 -(Methacryloyloxy) benzophenone in DMSO at concentrations of 3.16, 10.0, 31.6, 100, 316, 1000, 2500 and 5000.0 µg/plate in a plate incorporation assay in the presence and absence of mammalian metabolic activation (S9 mix).

4 -(Methacryloyloxy) benzophenone was tested up to cytotoxic concentrations. There was no evidence of induced mutant colonies over background.

The positive controls induced the appropriate responses in the corresponding strains.

This study is classified as acceptable and satisfies the requirement for Test Guideline OECD 471 for in vitro mutagenicity (bacterial reverse gene mutation) data.