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Toxicological information

Genetic toxicity: in vitro

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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:
Cyclic Hydrazides are mutagenic for Salmonella typhimurium
Author:
Errol Zeiger and Janet Guthrie
Year:
1981
Bibliographic source:
Mutation Research, 91 (1981) 199-205

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:
The Ames mutagenicity test was conducted on salmonella typhirium of strains TA98, TA 100 and TA 1535 to study the mutagenicity of chemical 1-amino-4-methylpiperazine.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): 4-Methylpiperazin-1-amine
- Molecular formula: C5H13N3
- Molecular weight: 115.179 g/mol
- Substance type: organic
- Physical state: Liquid
- Smiles notation: N1(CCN(CC1)C)N
- InChl: 1S/C5H13N3/c1-7-2-4-8(6)5-3-7/h2-6H2,1H3
Specific details on test material used for the study:
- Name of the test material: 4-methylpiperazin-1-amine
- IUPAC name: 4-methylpiperazin-1-amine
- Molecular weight: 115.179 g/mol
- Molecular formula: C5H13N3
- Substance type: Organic

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
other: TA98, TA100 and TA1535
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 fraction was prepared from Aroclor 1254-induced rat-liver extract
Test concentrations with justification for top dose:
0, 1, 3, 10, 30, 100 or 300 µmoles/plate
Vehicle / solvent:
No data
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
benzo(a)pyrene
other: 4-nitro-o-phenylene (TA98, without S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data
- Exposure duration: 2 days
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: All doses were run in triplicate with concurrent solvent controls, and all tests were repeated at least once

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data
Rationale for test conditions:
No data
Evaluation criteria:
The plates were observed for a dose dependent increase in the number of revertants/plate
Statistics:
Mean plate count ± S.E.M. (3 plates)

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
other: TA1535 and TA100
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
other: TA1535 and TA100
Metabolic activation:
without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
No data

Any other information on results incl. tables

TABLE 1: Mutagenicity of1-Amino-4-MethylpiperazineFor S. Typhimurium TA1535 In The Absence Of S9

Dose (µmoles/plate)

NPA

0

11±1

0.03

-

0.1

-

0.3

-

1

9±3

3

16±3

10

50±4

30

384±23

100

770±71

300

611±49

aMean plate count _+ S.E.M. (3 plates).

bNot done.

CToxic response; few or no his + revertants on plates.

Applicant's summary and conclusion

Conclusions:
1-amino-4-methylpiperazine did not show any mutagenic nature in Salmonella typhimurium strain TA98 in the presence and absence of S9 and also in strains TA1535 and TA100 in the presence of S9. However in the absence of S9, 1-amino-4-methylpiperazine showed mutagenic behaviour in strains TA1535 and TA100. Based on the observations made, the test chemical is not likely to classify as a gene mutant in vitro.
Executive summary:

Gene mutation toxicity study was performed for the test chemical 1-amino-4-methylpiperazine. The study was perfomed using Salmonella typhimurium strains TA1535, TA100 and TA98 both in the presence and absence of S9 metabolic activation system. The test chemical was dissolved in suitable solvent at doses of 0, 1, 3, 10, 30, 100 or 300 µmoles/plate. The plates were incubated for 2 days before the observation of revertant colonies. 1-amino-4-methylpiperazine did not show any mutagenic nature in Salmonella typhimurium strain TA98 in the presence and absence of S9 and also in strains TA1535 and TA100 in the presence of S9. However in the absence of S9, 1-amino-4-methylpiperazine showed mutagenic behaviour in strains TA1535 and TA100. Based on the observations made, the test chemical is not likely to classify as a gene mutant in vitro.