Tetrylammonium bromide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Gene mutation toxicity study of the test chemical

Author:

Docherty et al

Year:

2006

Bibliographic source:

Green Chem.

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Gene mutation toxicity study (Ames assay) was performed to determine the mutagenic nature of test chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Tetrylammonium bromide

EC Number:

200-769-4

EC Name:

Tetrylammonium bromide

Cas Number:

71-91-0

Molecular formula:

C8H20N.Br

IUPAC Name:

N,N,N-triethylethanaminium bromide

Test material form:

solid

Details on test material:

- Name of test material: Tetraethylammonium bromide (4E-ammBr)
- IUPAC name: N,N,N-triethylethanaminium bromide
- Molecular formula: C8H20NBr
- Molecular weight: 210.157 g/mol
- Substance type: Organic
- Physical state: White Crystalline Powder
-Batch No. L203711607

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA98 and TA100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

S9 rat liver enzyme (RLE) mix

Test concentrations with justification for top dose:

0.01, 1, 5 or 20 mg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: No data- Justification for choice of solvent/vehicle: No data

Controls

Untreated negative controls:

yes

Remarks:

Sterile distilled water and DMSO

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

sodium azide

other: 2-aminofluorene (TA98)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)DURATION- Preincubation period: No data- Exposure duration: 48 hrs- Expression time (cells in growth medium): 48 hrs- Selection time (if incubation with a selection agent): No data- Fixation time (start of exposure up to fixation or harvest of cells): No dataSELECTION AGENT (mutation assays): No dataSPINDLE INHIBITOR (cytogenetic assays): No dataSTAIN (for cytogenetic assays): No dataNUMBER OF REPLICATIONS: TriplicateNUMBER OF CELLS EVALUATED: No dataDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No dataOTHER EXAMINATIONS:- Determination of polyploidy: No data- Determination of endoreplication: No data- Other: No dataOTHER: No data

Rationale for test conditions:

No data

Evaluation criteria:

A chemical is considered to be mutagenic if the number of induced revertants is two or more times greater than the number of spontaneous revertants.(1) a significant relationship between the number of revertant colonies and dose concentration and (2) a two-fold or more increase in the number of revertant colonies over the number of spontaneous revertants seen in the control and (3) a Mutagenicity Index (MI) value greater than 2.0 or a Mutagenicity Activity Ratio (MAR) greater than 2.5.

Statistics:

No data

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA98 and TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks:

Both the positive control chemicals yielded 20–100 times more revertants than the number of spontaneous revertants, indicating a high level of mutagenicity

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS- Effects of pH: No data- Effects of osmolality: No data- Evaporation from medium: No data- Water solubility: No data- Precipitation: No data- Other confounding effects: No dataRANGE-FINDING/SCREENING STUDIES: Preliminary data was obtained from a rapid spot-test screening experiment indicating that the ILs tested were non-mutagenic at quantities below 1 mg/ plate.COMPARISON WITH HISTORICAL CONTROL DATA: No dataADDITIONAL INFORMATION ON CYTOTOXICITY: No data

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Salmonella typhimurium strain TA98 frameshift mutation results

Dose/mg/plate	Without S9					With S9
	No. revertants	MI	MAR	R^2	pvalue	No. revertants	MI	MAR	R^2	pvalue
0.01	21.00±1.00	0.91 ± 0.04	20.11	0.0536	0.469	27.67 ± 4.04	1.20 ± 0.18	0.19	0.1396	0.232
1	30.67 ± 6.43	1.33 ± 0.28	0.40			40.00 ± 6.56	1.74 ± 0.29	0.68
5	21.67 ± 4.16	0.94 ± 0.18	20.07			28.67 ± 2.52	1.25 ± 0.11	0.23
20	29.33 ± 14.01	1.28 ± 0.61	0.33			27.00 ± 4.58	1.17 ± 0.20	0.16
2-aminofluorene						3001.33 ± 528.91
DMSO	22.66 ± 2.08					30.67 ± 4.73
H2O	22.33 ± 6.11					26.00 ± 4.58
Spontaneous	22.67 ± 6.51					23.00 ± 8.19

 

Salmonella typhimurium strain TA100 frameshift mutation results

Dose/mg/plate	Without S9					With S9
	No. revertants	MI	MAR	R^2	pvalue	No. revertants	MI	MAR	R^2	pvalue
0.01	162.00±22.54	1.28 ± 0.18	0.25	0.2921	0.070	156.67 ± 7.02	1.21 ± 0.05	0.17	0.0707	0.403
1	131.67 ± 3.51	1.04 ± 0.03	0.03			164.00 ± 9.54	1.26 ± 0.07	0.22
5	163.00 ± 22.11	1.28 ± 0.17	0.26			153.67 ± 21.39	1.18 ± 0.16	0.15
20	179.33 ± 20.03	1.41 ± 0.16	0.38			151.67 ± 12.34	1.17 ± 0.09	0.14
Sodium azide	1817.33 ± 563.25
DMSO	112.33 ± 7.23					156.00 ± 6.24
H2O	135.33 ± 22.03					148.33 ± 9.07
Spontaneous	127.33 ± 15.50					129.67 ± 1.15

Applicant's summary and conclusion

Conclusions:

The test chemical did not induce mutation in Salmonella typhimurium strain TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.

Executive summary:

Gene mutation toxicity study (Ames assay) was performed to determine the mutagenic nature of test chemical. The study was performed using Salmonella typhimurium strain TA98 and TA100 with and without S9 metabolic activation system at dose levels of 0.01, 1, 5, 20 mg/plate. The doses were selected on the basisrapid spot-test screening experimentindicating that the test chemical wasnon-mutagenic at quantities below 1 mg/ plate. Sodium azide (TA100) and 2-aminofluorene (TA98) were used at positive controls and sterile distilled water and DMSO was used as negative controls. The plates were incubated for 48 hrs and the number of induced revertants was counted. The test chemical did not induce gene mutation in Salmonella typhimurium strain TA98 and TA100 in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.