[No public or meaningful name is available]

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other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Test guideline 3

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)

GLP compliance:

yes (incl. QA statement)

Remarks:

Experimental Toxicology and Ecology, BASF Aktiengesellschaft, 67056 Ludwigshafen, Germany

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 1-Hexanol, 2-ethyl-, reaction products with 1,6-diisocyanatohexane and Hexane, 1,6-diisocyanato-, homopolymer

EC Number:

939-549-4

Molecular formula:

Unspecified

IUPAC Name:

Reaction mass of 1-Hexanol, 2-ethyl-, reaction products with 1,6-diisocyanatohexane and Hexane, 1,6-diisocyanato-, homopolymer

Test material form:

other: Liquid

Details on test material:

- Storage stability: The stability of the test substance under storage conditions throughout the study period is guaranteed as indicated by the sponsor
- Date of production: 16 Nov 2005
- Physical state, appearance: Liquid, colorless to yellowish
- Storage conditions: Room temperature (N2 conditions)

Method

Target gene:

Salmonella typhimurium: Histidine
Escherichia coli: Tryptophan

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

The Salmonella strains are checked for the following characteristics at regular intervals: deep rough character (rfa); UV sensitivity (∆ uvrB); ampicillin resistance (R factor plasmid).
Histidine auxotrophy is automatically checked in each experiment via the spontaneous rate.

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

E. coli WP2 uvrA is checked for UV sensitivity.
Tryptophan auxotrophy is automatically checked in each experiment via the spontaneous rate.

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S-9 mix

Test concentrations with justification for top dose:

1st experiment, standard plate test: Salmonella strains and E. coIi WP2 uvrA with and without S-9 mix: 0, 20, 100, 500, 2500, and 5000 µg/plate
2nd experiment, standard plate test: Salmonella strains and E. coIi WP2 uvrA with and without S-9 mix: 0, 0.8, 4, 20, 100 and 500 µg/plate
3rd experiment, preincubation test: Salmonella strains without S-9 mix 0, 0.8, 4, 20, 100 and 500 µg/plate; Salmonella strains with S-9 mix 0, 4, 20, 100, 500, and 2 500 µg/plate; E. coIi WP2 uvrA with and without S-9 mix 0, 4, 20, 100, 500, and 2500 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Due to the Iimited 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

Controls 1

Untreated negative controls:

yes

Remarks:

Additional plates are treated with soft agar, S-9 mix, buffer, vehicle or the test substance but without the addition of tester strains

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

Strain: E. coli WP2 uvrA, Without S9 mix

Controls 2

Untreated negative controls:

yes

Remarks:

Additional plates are treated with soft agar, S-9 mix, buffer, vehicle or the test substance but without the addition of tester strains

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

Strain: TA 1537, Without S-9 mix

Controls 3

Untreated negative controls:

yes

Remarks:

Additional plates are treated with soft agar, S-9 mix, buffer, vehicle or the test substance but without the addition of tester strains

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylendiamine

Remarks:

Strain: TA 98, Without S-9 mix

Controls 4

Untreated negative controls:

yes

Remarks:

Additional plates are treated with soft agar, S-9 mix, buffer, vehicle or the test substance but without the addition of tester strains

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: N-methyl-N'-nitro-N-nitrosoguanidine

Remarks:

Strain: TA 1535, TA 100, Without S-9 mix

Controls 5

Untreated negative controls:

yes

Remarks:

Additional plates are treated with soft agar, S-9 mix, buffer, vehicle or the test substance but without the addition of tester strains

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

Strain TA 1535, TA 100, TA 1537, TA 98 and E. coli WP2 uvrA, with S-9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation;

DURATION
Standard plate test
- Exposure duration: 48-72 hours at 37°C in the dark
Preincubation test
- Preincubation period: 20 minutes at 37°C
- Exposure duration: 48-72 hours at 37°C in the dark

NUMBER OF REPLICATIONS: 3 test plates per dose or per control

DETERMINATION OF CYTOTOXICITY
- Toxicity detected by a decrease in the number of revertants, clearing or diminution of the background lawn (= reduced his- or trp- background growth), and reduction in the titer

Evaluation criteria:

- Generally, the experiment is considered valid if the following criteria are met: (1) The number of revertant colonies in the negative controls was within the normal range of the historical control data for each tester strain, (2) The sterility controls revealed no indication of bacterial contamination, (3) The positive control articles both with and without S-9 mix induced a significant increase in the number of revertant colonies within the range of the historical control data or above, (4) The titer of viable bacteria was ≥ 1E8/mL.
- The test chemical is considered positive in this assay if the following criteria is 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

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A bacteriotoxic effect was observed depending on the strain and test conditions from about 500 µg - 2 500 µg/plate onward.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

- Toxicity: A bacteriotoxic effect (reduced background growth, decrease in the number of revertants, reduction in the titer) was observed depending on the strain and test conditions from about 500 - 2500 µg/plate onward.
- Solubility: Test substance precipitation was found from about 2500 µg/plate onward.
- Mutagenicity: An increase in the number of his+ or trp+ revertants was not observed in the standard plate test or in the preincubation test either without S-9 mix or after the addition of a metabolizing system.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.