Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
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

Reference
Name:
Unnamed
Type:
Constituent
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

R2

pvalue

No. revertants

MI

MAR

R2

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

R2

pvalue

No. revertants

MI

MAR

R2

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.