Acetic anhydride

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:

other: Comparable to guideline with acceptable restrictions (limited reporting)

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Assay of 855 test chemicals in ten tester strains using a new modification of the Ames test for bacterial mutagens.

Author:

McMahon RE et al.

Year:

1979

Bibliographic source:

Cancer Res 39: 682-693.

Reference 2

Reference Type:

secondary source

Title:

Acetic Anhydride

Author:

OECD SIDS

Year:

1997

Bibliographic source:

SIDS Initial Assessment Report for 6th SIAM (Paris, 9-11 June 1997)

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:

As described in Cline, J.C. and R.E. McMahon. Detection of chemical mutagens. Use of concentration gradient plates in a High Capacity Screen. Res. Commun. Chem. Pathol. Pharmacol. 16.523, 1977.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Acetic anhydride

EC Number:

203-564-8

EC Name:

Acetic anhydride

Cas Number:

108-24-7

Molecular formula:

C4H6O3

IUPAC Name:

acetyl acetate

Details on test material:

- Name of test material (as cited in study report): acetic anhydride

Method

Target gene:

histidine and tryptophan auxotrophy

Species / strain

Species / strain / cell type:

other: TA1535, TA100, TA1537, TA1538, TA98, E. coli WP2 uvrA-

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S-9 mix

Test concentrations with justification for top dose:

1-10000 µg/plate (0.1-1000 µg/mL)

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

with S9 mix

Controls 2

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

without S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: Agar plates containing a concentration gradient of test compound were prepared as follows. Ten mL of minimal agar medium (not containing test compound) were poured into a square Petri dish (9x9 cm) which was tilted at a slight angle. The agar was then allowed to solidify into a wedge-shaped layer by standing at room temperature. Meanwhile, a 1000 µg/mL mixture of test compound in agar was prepared by adding 10 mL of minimal agar to 0.1 mL of a 100 mg/mL solution of test compound in dimethyl sulfoxide. When appropriate, water or demethoxyethane was used instead of dimethyl sulfoxide. The cooled agar plates were then placed on a level surface, and an overlay of the 10 mL of agar containing the test compound was poured onto the plate to form a reversed wedge of agar on top of the first wedge. A concentration gradient of compound was produced by allowing the compound in the upper wedge to diffuse into the lower layer for 2 hr at room temperature. The concentration range in this plate was approximately 100 to 1000 µg/mL. Three additional plates with concentration ranges of 10 to 100 µg/mL, 1 to 10 µg/mL, and 0.1 to 1 µg/mL were prepared. A streaking device consisting of 10 sterile 50 µL pipettes was next dipped into suspensions of the 10 test strains and allowed to fill by capillary action. The pipettes were then touched to the upper edge of the gradient and drawn across the plate. A wet trail of inoculums was observed. The plates are then incubated for 48 h at 37 °C. The minimal medium used in these studies was supplemented with tryptophan, histidine, and biotin in amounts suffiecient to allow growth of only about 6 to 10 generations of auxotroph, which would allow expression of mutagenic events. If no mutagenic events occur, a very pale streak of bacterial growth was seen along the inoculation streak. When nonlethal mutagnic events occur, discrete colonies appear in this pale background lawn. The frequency of colony appearance increases along the increasing gradient to a concentration at which maximal mutation occurs. Conversely, frequency decreases along the decreasing gradient to a concentration below which only background lawn was observable. These upper and lower concetrations were reported as the concentration range in which the compound was mutagenic under test conditions. A second effect of a test compound, cytotoxicity, was also observed with many compounds. The concentration range over which the test compound was too toxic to permit growth of the auxotroph can be observed by the absence of growth along the application streak (i.e. no backround lawn is observable).

Results and discussion

Test results

Species / strain:

other: TA1535, TA100, TA1537, TA1538, TA98, E. coli WP2 uvrA-

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: other: TA1535, TA100, TA1537, TA1538, TA98, E. coli WP2 uvrA-

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

An increase in revertants was not observed in any strain of Salmonella typhimurium or E coli. Therefore based on the results, the test substance does not require any classification based on EU and GHS standards.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative