Hexylamine

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(May 1983/Revised Draft, March 1996)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(May 1983/Revised Draft, March 1996)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(December 1992)

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:

Hexylamine

EC Number:

203-851-8

EC Name:

Hexylamine

Cas Number:

111-26-2

Molecular formula:

C6H15N

IUPAC Name:

hexan-1-amine

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:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

20 ; 100 ; 500 ; 2,500 and 5,000 μg/plate with and without metabolic activation (standard plate test)
125 ; 250 ; 500 ; 1,000 and 2,000 μg/plate with and without metabolic activation (preincubation test)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO;
- Justification for choice of solvent/vehicle: Due to the limited solubility of the test substance in water, DMSO was selected as the vehicle, which had been demonstrated to be suitable in bacterial reverse mutation tests and for which historical control data are available .

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S9: 2-aminoanthracene (2-AA); without S9: N-methyl-N' -nitro-N-nitrosoguariidine (MNNG), 4-nitro-o-phenylendiamine (NOPD), 9-aminoacridine, N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG)

Details on test system and experimental conditions:

METHOD OF APPLICATION: Standard plate incorporation and preincubation test

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 - 72 hours

NUMBER OF REPLICATIONS: triplicates each in two independent experiments

DETERMINATION OF CYTOTOXICITY
- Method:
• decrease in the number of revertants
• clearing or diminution of the background lawn (= reduced his or trp background growth)
• reduction in the titer

OTHER:
Positive controls:
with S9:
- 2-aminoanthracene (2-AA);
• 2.5 µg/plate in DMSO for TA1535, TA100, TA1537, TA98;
• 60 µg/plate in DMSO for E. coli WP2 uvrA;
without S9:
- N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)
• 5 µg/plate in DMSO for TA1535, TA100;
- 4-nitro-o-phenylendiamine (NOPD)
• 10 µg/plate in DMSO for TA98;
- 9-aminoacridine (AAC)
• 100 µg/plate in DMSO for TA1537;
- N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG)
• 10 µg/plate in DMSO for E. coli WP2 uvrA;

To demonstrate the efficacy of the S9 mix in the assay, the S9 batch was characterized with benzo(a)pyrene.

Evaluation criteria:

The test chemical is considered positive in this assay if the following criteria are met:
A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.

A test substance is generally considered nonmutagenic in this test if:
The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

(No increase in the number of his-positive revertants)

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A bacteriotoxic effect was observed under all test conditions.

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

(No increase in the number of trp-positive revertants)

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A bacteriotoxic effect was observed under all test conditions.

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No test substance precipitation was found.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
- A bacteriotoxic effect (reduced his-negative or trp-negative background growth, decrease in the number of his-positive or trp-postive revertants, reduction in the titer) was observed in the standard plate test from about 2500 µg/plate onward.
- In the preincubation assay bacteriotoxicity was found from about 500 - 1,000 μg/plate onward .

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.