Tripiperazine dicitrate

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity Tests On Anthelmintics: Microsomal activation of Viprynium Embonate to a mutagen

Author:

D.G. Macphee and D.M. Podger

Year:

1977

Bibliographic source:

Mutation Research, 48 (1977) 307-312

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as metioned below

Principles of method if other than guideline:

A slightly modified version of the procedure recommended by Ames et al. was adopted, in that the test samples were simply placed in "wells" cut out of the agar of a plate previously seeded with the appropriate tester strain.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Tripiperazine dicitrate

EC Number:

205-622-8

EC Name:

Tripiperazine dicitrate

Cas Number:

144-29-6

Molecular formula:

C6H8O7.3/2C4H10N2

IUPAC Name:

piperazine 2-hydroxypropane-1,2,3-tricarboxylate (3:2) (salt)

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Piperazine Citrate - Molecular formula (if other than submission substance): C12H30N6•Cl2H16O14 - Molecular weight (if other than submission substance): 642.76 g/mole - Smiles notation (if other than submission substance): C1CNCCN1.C1CNCCN1.C1CNCCN1.C(C(=O)O)C(CC(=O)O)(C(=O)O)O.C(C(=O)O)C(CC(=O)O)(C(=O)O)O - Substance type: Organic - Physical state: Solid-Purity:99.0% (BP)

Specific details on test material used for the study:

- Name of test material (as cited in study report): Piperazine citrate / tripiperazine dicitrate- Molecular formula: C12H30N6•Cl2H16O14 - Molecular weight: 642.76 g/mole - Substance type: Organic - Physical state: Solid

Method

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

not specified

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix, which contained per ml: 0.3 ml of S-9 fraction 8 µmol MgCl2, 33 µmol KCl, 5 µmol glucose-6-phosphate, 4 µmol NADP, and 100 µmol sodium phosphate

Test concentrations with justification for top dose:

5000 µg/ml or 50,000 µg/ml.

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)DURATION- Preincubation period: 2-3 days- Exposure duration: 24 hrs- Expression time (cells in growth medium): not specified- Selection time (if incubation with a selection agent): not specified- Fixation time (start of exposure up to fixation or harvest of cells): not specifiedSELECTION AGENT (mutation assays): SPINDLE INHIBITOR (cytogenetic assays): not specifiedSTAIN (for cytogenetic assays): not specifiedNUMBER OF REPLICATIONS: not specifiedNUMBER OF CELLS EVALUATED: not specifiedDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other:OTHER EXAMINATIONS: not specified- Determination of polyploidy:- Determination of endoreplication:- Other:

Rationale for test conditions:

not specified

Evaluation criteria:

not specified

Statistics:

not specified

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA98, TA100, TA1535 and TA1538TA98, TA100, TA1535 and TA1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

not specified

Applicant's summary and conclusion

Conclusions:

The test substance piperazine citrate gave negative results for mutagenicity with or without S9 metabolic activation system when tested at concentration of 5000 µg/ml or 50,000 µg/ml in Salmonella typhimurium TA98, TA100, TA1535 and TA1538. Thus the test chemical is not likely to classify as a gene mutant in vitro as per the CLP classification criteria.

Executive summary:

The test substance piperazine citrate was tested for mutagenicity in the Salmonella typhimurium test system. The strains used wereTA98, TA100, TA1535 and TA1538.The test concentrations used were 5000µg/ml or 50,000µg/ml.A slightly modified version of the procedure recommended by Ames et al. was adopted, in that the test samples were simply placed in "wells" cut out of the agar of a plate previously seeded with the appropriate tester strain. Addition of a mixture of rat liver microsomal enzymes and appropriate co-factors ("S-9 mix") to one of the two wells on a single plate allowed a possible requirement for metabolic activation to be recognised. Using this procedure, the test chemical was determined to be non-mutagenic in this system. Controls with DMSO alone and S-9 mix alone also gave negative results. Thus the test chemical is not likely to classify as a gene mutant in vitro as per the CLP classification criteria.