Butanal, reaction products with aniline

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: scientifically acceptable and well documented

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Principles of method if other than guideline:

The condensation product of acroleins with aromatic bases was examined for mutagenic activity in four histidine-dependent auxotrophs of Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537, using pour-plate assays. The procedures used complied with the OECD Guidelines for testing of chemicals (1983) and the EPA Toxic Substances Control Act Test Guidelines (1985). Each test, in each strain, was conducted twice.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Butanal, reaction products with aniline

EC Number:

270-109-8

EC Name:

Butanal, reaction products with aniline

Cas Number:

68411-20-1

IUPAC Name:

aniline; butanal

Test material form:

other: reddish-brown liquid

Details on test material:

The condensation product of acroleins with aromatic bases (also referred to as ACAB) , a reddish-brown liquid, was received from WTR. The material was further identified by the lot no. 0 E 30616, WTR NS' 32 and CAS Reg. No. 68411-20-1, and had a density of 0.98 9/cm. It was stored in the
original container at room temperature and protected from light.

Method

Target gene:

no data

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

Cultures of the histidine-dependent strains of Salmonella typhimurium were derived from cultures provided by Prof. Bruce Ames, University of California. The characteristics of the indiVidual strains are as follows:
TA 1535 - contains a histidine missense mutation but is also deficient in a DNA repair system (uvr B) and has a defective lipopolysaccharide coat on the cell wall. It is reverted by many agents causing base-pair substitutions, but is not sensitive to frameshift mutagens.
TA 100 - is the same as TA 1535 but contains a resistance transfer factor conferring ampicillin resistance and increasing sensitivity to some mutagens (plasmid pKM 101). In addition to base-pair substitutions, it is also able to detect certain frameshift mutagens.
TA 1537 - bears a histidine frameshift mutation. Like TA 1535, it is defective in a DNA repair system and lipopolysaccharide coat. It is sensitive to agents causing frameshift mutations involving insertion or deletion of a single base-pair.
TA 98 - contains another histidine frameshift mutation. Again it has a defective DNA repair system and lipopolysaccharide coat but also contains the pKM 101 plasmid. It is reverted by agents causing deletion of two adjacent base-pairs (double frameshift mutations), but not by simple alkylating agents causing base-pair substitutions.

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix

Test concentrations with justification for top dose:

Test 1: 50, 158, 500, 1580, 5000 µg/plate
Test 2: 50, 158, 500, 1580, 5000 µg/plate

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

benzo(a)pyrene

other: 2-aminoanthracene

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

Any other information on results incl. tables

The studies, which were conducted in the absence and presence of an activating system derived from rat liver (S-9 mix), employed a

range of levels of the test material from 50 to 5000 ug per plate, selected following a preliminary toxicity test in strain TA 98.

All tests included solvent (dimethyl sulphoxide) controls with and without S-9 mix.

No increases in reversion to prototrophy were obtained with any of the four bacterial strains at the test material levels tested, either in the presence or absence of S-9 mix.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative

Executive summary:

The condensation product of acroleins with aromatic bases was examined for mutagenic activity in four histidine-dependent auxotrophs of Salmonella typhimurium, strains TA 98, TA 100, TA 1535 and TA 1537, using pour-plate assays. The procedures used complied with the OECD Guidelines for testing of chemicals (1983) and the EPA Toxic Substances Control Act Test Guidelines (1985).

Each test, in each strain, was conducted on two separate occasions.

The studies, which were conducted in the absence and presence of an activating system derived from rat liver (S-9 mix), employed a range of levels of the test material from 50 to 5000 ug per plate, selected following a preliminary toxicity test in strain TA 98.

All tests included solvent (dimethyl sulphoxide) controls with and without S-9 mix.

No increases in reversion to prototrophy were obtained with any of the four bacterial strains at the test material levels tested, either in the presence or absence of S-9 mix.

Marked increases in the number of revertant colonies were induced by the known mutagens benzo[a]pyrene, 2-nitrofluorene, 2-aminoanthracene, 9-aminoacridine and sodium azide when examined under similar conditions.

It was concluded that the condensation product of acroleins with aromatic bases was devoid of mutagenic activity under the conditions of the test.