Reaction mass of 3-methyl-1-octylpyridinium chloride and 4-methyl-1-octylpyridinium chloride

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 guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

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:

Reaction mass of 3-methyl-1-octylpyridinium chloride and 4-methyl-1-octylpyridinium chloride

EC Number:

941-484-1

Molecular formula:

not possible

IUPAC Name:

Reaction mass of 3-methyl-1-octylpyridinium chloride and 4-methyl-1-octylpyridinium chloride

Test material form:

liquid: viscous

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Details on mammalian cell type (if applicable):

TA 98: his D 3052; rfa-; uvrB-; R-factor: frame shift mutations
TA 100: his G 46; rfa-; uvrB-; R-factor: base-pair substitutions
TA 1535: his G 46; rfa-; uvrB-: base-pair substitutions
TA 1537: his C 3076; rfa-; uvrB-: frame shift mutations
TA 102: his G 428 (pAQ1); rfa-; R-factor: base-pair substitutions

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (rat)

Test concentrations with justification for top dose:

Experiment I:
3.16, 10.0, 31.6, 100, 316, 1000, 2500 and 5000 μg/plate
(for TA 98 with and without metabolic activation and TA 100 with metabolic activation)
0.316, 1.00, 3.16, 10.0, 31.6, 100, 316, 1000, 2500 and 5000 μg/plate
(for TA 100 without metabolic activation and all other tester strains with and without metabolic activation)

Experiment II:
0.100, 0.316, 1.00, 3.16, 10.0, 31.6, 100, 316, 1000 and 2500 μg/plate

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

dest. water

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Remarks:

Negative/solvent controls (A. dest.) and solvent controls (DMSO) were treated in the same way as all dose groups.

Controls 2

Positive controls:

yes

Remarks:

10 μg/plate

Positive control substance:

sodium azide

Remarks:

TA 100, TA 1535 Without metabolic activation

Controls 3

Positive controls:

yes

Remarks:

10 μg/plate for TA 98, 40 μg/plate for TA 1537

Positive control substance:

other: 4-nitro-o-phenylene-diamine

Remarks:

TA 98, TA 1537 Without metabolic activation

Controls 4

Positive controls:

yes

Remarks:

1 μL/plate

Positive control substance:

methylmethanesulfonate

Remarks:

TA 102 Without metabolic activation

Controls 5

Positive controls:

yes

Remarks:

2.5 μg/plate; 10 μg/plate for TA 102

Positive control substance:

other: 2-aminoanthracene

Remarks:

TA 98, TA 100, TA 1535, TA 1537 and TA 102

Details on test system and experimental conditions:

METHOD OF APPLICATION: 1st main experiment: in agar (plate incorporation); 2nd main experiment: preincubation

DURATION
- 1st main experiment: 48 hours incubation (37°C)
- 2nd main experiment: 60 min preincubation (37°C), 48 hours incubation (37°C)

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: determination of background lawn, reduction in the number of revertants down to a mutation factor of approximately ≤ 0.5 in relation to the solvent control

Evaluation criteria:

A test is considered acceptable if for each strain:
- the bacteria demonstrate their typical responses to ampicillin (TA 98, TA 100, TA 102)
- the control plates with and without S9 mix are within the historical control data range
- corresponding background growth on negative control, solvent control and test plates is observed
- the positive controls show a distinct enhancement of revertant rates over the control plate

A test item is considered as mutagenic if:
- a clear and dose-related increase in the number of revertants occurs and/or
- a biologically relevant positive response for at least one of the dose groups occurs in at least one tester strain with or without metabolic activation.
A biologically relevant increase is described as follows:
- if in tester strains TA 98, TA 100 and TA 102 the number of reversions is at least twice as high
- if in tester strains TA 1535 and TA 1537 the number of reversions is at least three times higher than the reversion rate of the solvent control.

A test item producing neither a dose related increase in the number of revertants nor a reproducible biologically relevant positive response at any of the dose groups is considered to be non-mutagenic in this system.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not examined

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

No precipitation of the test item was observed in any tester strain used in experiment I and II (with and without metabolic activation).

Toxic effects of the test item were noted in all tester strains evaluated in experiment I and II.

In experiment I toxic effects of the test item were observed in tester strains TA 100, TA 1535, TA 1537 and TA 102 at concentrations of 316 μg/plate and higher (without metabolic activation). In tester strain TA 98 toxic effects of the test item were observed at concentrations of 1000 μg/plate and higher (without metabolic activation) and in tester strains TA 100 and TA 1535 (with metabolic activation). Also, in tester strains TA 98, TA 1537 and TA 102 toxic effects of the test item were noted at concentrations of 2500 μg/plate and higher (with metabolic activation).

In experiment II toxic effects of the test item were noted in all tester strains at concentrations of 100 μg/plate and higher (without metabolic activation).
In tester strain TA 100 toxic effects of the test item were observed at concentrations of 316 μg/plate and higher (with metabolic activation) and in tester strains TA 98, TA 1535, TA 1537 and TA 102 toxic effects of the test item were noted at concentrations of 1000 μg/plate and higher (with metabolic activation).

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative without metabolic activation
negative with metabolic activation

Under the conditions of this test the susbstance did not cause gene mutations by base pair changes or frameshifts in the genome of the tester strains used. Therefore, the substance is considered to be non-mutagenic in this bacterial reverse mutation assay.