2-nitrophenyl acetate

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)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

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:

Acetic acid, 2-nitrophenyl ester

IUPAC Name:

Acetic acid, 2-nitrophenyl ester

Constituent 2

Reference substance name:

2-nitrophenyl acetate

EC Number:

210-233-1

EC Name:

2-nitrophenyl acetate

Cas Number:

610-69-5

Molecular formula:

C8H7NO4

IUPAC Name:

2-nitrophenyl acetate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): 2-Nitrophenyl Acetate

- Physical state: solid light yellow
- Analytical purity: 95 % w/w

- Impurities (identity and concentrations):
Phenol <5.0 % (w/w)
2,4-dinitrophenol <5.0 % (w/w)
2,6-dinitrophenol <5.0 % (w/w)
Acetic acid <1.0 % (w/w)
Water <5.0 % (w/w)
2-nitrophenol <5.0 % (w/w)

- Lot/batch No.: 201408
- Expiration date of the lot/batch: unlisted

- Storage condition of test material: The test substance will be stored in dry room in dark in closed container at the room temperature

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction of rat liver homogenate and mixture of cofactors

Test concentrations with justification for top dose:

The doses used were 5, 15, 50, 125 and 250 μg per plate.

Vehicle / solvent:

Solvent: Dimethyl sulfoxide p.a., Merck, Lot. No.: K45609252 424, exp. 05/2017

Controls

Untreated negative controls:

yes

Remarks:

solvent control

Negative solvent / vehicle controls:

yes

True negative controls:

yes

Remarks:

untreated control (no solvent)

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine (NPD) CAS No.: 99-56-9, Aldrich; 2-aminoanthracene (2-AA) CAS No: 613-13-8, Sigma; N-methyl-N´-nitro-N-nitrosoguanidine (MNNG) CAS No.: 70-25-7, TCI Europe

Evaluation criteria:

The main criterion for evaluation of results was modified two-fold increase rule, which is compatible with the application of statistical methods (2, 3). After this rule the result is positive, if a reproducible dose-response effect occurs and/or a doubling of the ratio Rt/Rc is reached.
An increase is considered as „biologically relevant“:
- if the number of reversions is at least twice as high as that in the solvent control for the strains having spontaneous reversion >10;
- if the number of reversions is at least three times as high as that in the solvent control for the strains having spontaneous reversion ≤10;
A test substance producing neither a dose-related increase in the number of revertants nor a reproducible biologically relevant positive response at any of the dose groups is considered to be non-mutagenic in this system.
According to OECD TG 471, the biological relevance of the results is the criterion for the interpretation of results, a statistical evaluation of the results is not regarded as necessary.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA:

Spontaneous reversions in negative and positive controls were compared with historical controls in our laboratory. The current ranges are given in the Table A

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table A: Current historical ranges of revertant numbers in bacterial strains used in the study and live bacteria count used in the first experiments

Control	Spont.rev.	DMSO	PC -S9	PC +S9	Number ofCFU/mL
Strain
S.t. TA 100	77-181	70-174	320-824	548-1568	1.05*10^9
S.t. TA 1535	9-29	10-26	258-626	68-260	1.09*10^9
S.t. TA 98	13-49	12-44	282-1550	545-2209	3.63*10^8
S.t. TA 1537	5-21	4-20	461-1401	61-281	1.93*10^8
E.c. WP2 uvrA	12-48	8-40	379-1103	0-387	1.24*10^9

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the above-described experimental design, the test substance 2-Nitrophenyl acetate was non-mutagenic for all strains Salmonella typhimurium TA 100, TA 98, TA 1535, TA 1537 and Escherichia coli WP2 uvrA in experiments with as well as without metabolic activation.

Executive summary:

The test substance 2-Nitrophenyl Acetate was assayed for the mutagenicity by the Bacterial Reverse Mutation Test. The performed test was based on EU method B.13/14 Mutagenicity – Reverse mutation test using bacteria, which is analogous to the OECD Test Guideline No. 471. Four indicator Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and one indicator Escherichia coli WP2 uvrA strain were used. The test substance was dissolved in dimethylsulfoxide and assayed in doses of 5-250 (500) microg per plate, which were applied to plates in volume of 0.1 mL. Experiments were performed without as well as with metabolic activation with a supernatant of rat liver and a mixture of cofactors. In the arrangement given above, the test substance 2-Nitrophenyl Acetate was non-mutagenic for all strains Salmonella typhimurium TA 100, Ta 98, TA 1535, TA 1537 and Escherichia coli WP2 uvrA in experiments with as well as without metabolic activation.