p-[(2-chloroethyl)ethylamino]benzaldehyde

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: Study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1980

Report date:

1980

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Principles of method if other than guideline:

Method: other according to Ames et al., Mut. Res. 31, 347-364, 1975

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

p-[(2-chloroethyl)ethylamino]benzaldehyde

EC Number:

220-150-2

EC Name:

p-[(2-chloroethyl)ethylamino]benzaldehyde

Cas Number:

2643-07-4

Molecular formula:

C11H14ClNO

IUPAC Name:

4-[(2-chloroethyl)(ethyl)amino]benzaldehyde

Details on test material:

N-Ethyl-N-(2-chlorethyl)-p-amino-benzaldehyd (purity >90%)

Method

Species / strain

Species / strain / cell type:

other: Salmonella tyhpimurium TA 100, TA 1537, TA 98; E.coli WP2 uvrA

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix prepared from sprangue Dawlay rats after Aroclor 1254 activation

Test concentrations with justification for top dose:

5, 15.8, 50, 158, 500, 1580, 5000 µg/plate

Details on test system and experimental conditions:

The mutagenicity experiments with his- S. typhimurium were performed with minor modification of the method described by Ames. The test compound, 500 µl S-9 Mix from Aroclor1254-treated rats (or 500 µl 150 mM KCl), 100 µl of the bacterial suspension and 2000 µl top agar, which consisted of 0.55 % agar, 0.55 % NaCl, 50 µM histidine, 50 µM biotin, and 25 µM Na phosphate pH 7.4, 45°C, were mixed in a test tube and poured onto a
petridish with minimal agar consisting of 1.5 % agar and Vogel-Bonner E medium with 2 % glucose. Where indicated the epoxide hydratase inhibitor
and glutathione depletor, TCPO (2.6 µMol per plate) was added in 10 µl DMSO. Corresponding plates without DMSO received 10 µl DMSO. After
incubation in the dark for 3 days colonies (his+ revertants) were counted.
Expeximents with trp- E. coli were performed the same way, but histidine and biotin of the top agar were replaced by 50 µM tryptophan. Six different concentrations from 15.8 to 5000 µg were used in the experiments for direct mutagenicity. In the experiments with S-9 Mix, seven doses from 5 to 5000 µg were tested.
Every experiment contained positive controls for checking the activity of the metabolizing systemn and the mutability of the bacteria as well as negative controls in the form of sterility controls and incubations without test compound. As positive controls N-metyl-N-nitro-N-nitrosoguanidine and benzo(a)pyrene-4,5-oxide were used in the experiments for directmutagenicity and 3-methylcholanthrene, benzo(a)pyrene and 2-aminoanthracen
in the standard experiments with an activating system.

Results and discussion

Test results

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

other: 5000 µg/plate (without metabolic activation)

Additional information on results:

At doses >= 1580 µg, a precipitate, which was still present at the end of the experiment, was found on the plates. In the abscence of S-9 Mix, at 5000 µg practically all bacteria were killed. Under all other conditions, no significant decrease in survival occurred. Thus, adequate mutagenicity testing could be performed up to quite high doses. In the direct mutagenicity test, the test substance was mutagenic with TA 100, but not with the other strains. In the presence of S-9 Mix, the mutagenicity with TA 100 was potentiated and the compound became also mutagenic with WP2 uvrA and slightly with TA 98. The epoxide hydratase inhibitor and glutathione depletor 1,1,1 -trichloropropene oxide did not show any effect upon the mutagenicity of the test substance.

Remarks on result:

other: other: Salmonella tyhpimurium TA 100, TA 1537, TA 98; E.coli WP2 uvrA

Remarks:

Migrated from field 'Test system'.