Trientine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study followed OECD 471 guideline and the GLP Regulations. No significant deviations can be observed from the study guidelines; however, strain E. coli WP2 uvrA, or E. coli WP2 uvrA (pKM101), or S. typhimurium TA102 were not included. No data on purity.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Trientine

EC Number:

203-950-6

EC Name:

Trientine

Cas Number:

112-24-3

Molecular formula:

C6H18N4

IUPAC Name:

N,N'-bis(2-aminoethyl)ethane-1,2-diamine

Details on test material:

- Name of test material (as cited in study report): 6933-6-20
- Physical state: clear yellow liquid
- Analytical purity: no data
- Impurities (identity and concentrations):
- Lot/batch No.:# 92-014
- Stability under test conditions: no apparent change in the physical state of the test article during storage
- Storage condition of test material: at the room temperature in the clear glass container
- Other:

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

not applicable

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Details on mammalian cell type (if applicable):

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S-9 fraction of rat liver homogenate obtained from Arcolor 1254-treated Sprague Dawley rats

Test concentrations with justification for top dose:

16.7, 50, 167, 500, 1000 and 1670 µg/plate

Vehicle / solvent:

-solvent(s) used: DMSO;
- Justification for choice of solvent: Maron, D.M., J. Katzenellenbogen and B.N. Ames (1981) Compatibility of organic solvents with the Salmonella/Microsome Test, Mutation m., 88:343-350.

Controls

Controls 1

Negative solvent / vehicle controls:

yes

Remarks:

with and without metabolic activation

True negative controls:

no

Positive controls:

yes

Remarks:

without metabolic activation

Positive control substance:

sodium azide

Remarks:

TA1535 and TA100= 10 µg/plate

Controls 2

Negative solvent / vehicle controls:

yes

Remarks:

with and without metabolic activation

True negative controls:

no

Positive controls:

yes

Remarks:

without metabolic activation

Positive control substance:

9-aminoacridine

Remarks:

TA1537=150 µg/plate

Controls 3

Negative solvent / vehicle controls:

yes

Remarks:

with and without metabolic activation

True negative controls:

no

Positive controls:

yes

Remarks:

without metabolic activation

Positive control substance:

2-nitrofluorene

Remarks:

TA1538 and TA98= 5 µg/plate

Controls 4

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

with metabolic activation

Positive control substance:

other: 2-anthramine

Remarks:

in all tester strains=2.5 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: 48h
- Exposure duration: 48h



SELECTION AGENT (mutation assays): biotin and histidine



NUMBER OF REPLICATIONS:
test and control articles (positive and negative): triplicates of all five tested strains
test article in the prescreening: in duplicate



DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition is tested at the following concentrations: 50, 167, 500, 1670 and 5000 µg/plate

Evaluation criteria:

revertant colonies are counted (Artek electronic colony counter interfaced with an IBM PC/AT computer for data acquisation)
-positive result is defined as a statistically significant, dose-dependent increase in the number of histidine-independent revertans with at least one dose level inducing solvent control value
-negative results is defined as the absence of a statistically significant or dose-dependent increase in the number of histidine-independent revertants
-equivocal result is defined when the test article dose not induce a statistically significant, dose dependent increase in revertant frequency, but does induce a revertant frequency at one dose level that is two-fold the spontaneous control value

Statistics:

Statistical analyses were performed using the program developed by Snee and Irr (1981), with significance established at the 95% confidence limit.

Results and discussion

Test results

Test results 1

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not determined

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

not determined

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
- Growth inhibition was observed in strain TA1538 and TA100 at doses of 1670 and 5000 µg/plate without metabolic activation.
- Growth inhibition was observed in strain TA1535, TA1537, TA1538, TA98, TA100 at doses 500, 1000 and /or 1670 µg/plate with and /or without metabolic activation.
- No increase (compared to the negtive control cultures) in revertant frequencies was seen in strain TA1538.
- A dose-dependent increase in revertant frequencies to approx 1.9- to 13- fold control values, were observed in strain TA1538, TA1537, TA98 and TA100 without metabolic activation.


COMPARISON WITH HISTORICAL CONTROL DATA: no data

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive with and without metabolic activation

The results for TETA were positive in the Ames/Salmonella Plate Incorporation Assay under the conditions, and according to, the criteria, of the test protocol.

Executive summary:

Triethylenetetramine (TETA) was tested for potential mutagenic activity using the Salmonella/microsome bacterial mutagenicity assay (Ames test).

In the RANGE-FINDING/SCREENING STUDIES Growth inhibition was observed in strain TA1538 and TA100 at doses of 1670 and 5000 µg/plate without metabolic activation. Growth inhibition was observed in strain TA1535, TA1537, TA1538, TA98, TA100 at doses 500, 1000 and /or 1670 µg/plate with and /or without metabolic activation.

No increase (compared to the negtive control cultures) in revertant frequencies was seen in strain TA1538. A dose-dependent increase in revertant frequencies to approx 1.9- to 13- fold control values, were observed in strain TA1538, TA1537, TA98 and TA100 without metabolic activation. Thus, TETA was considered to be mutagenic in this in vitro bacterial assay.