4-methylpiperazin-1-amine

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:

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

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

Test material information

Constituent 1

Reference substance name:

4-methylpiperazin-1-amine

EC Number:

230-053-7

EC Name:

4-methylpiperazin-1-amine

Cas Number:

6928-85-4

Molecular formula:

C5H13N3

IUPAC Name:

4-methylpiperazin-1-amine

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:

S. typhimurium, 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 results

Test results 1

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

Test results 2

Species / strain:

S. typhimurium, 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

Test results 3

Species / strain:

S. typhimurium, 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)	NP^A
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.