1-Piperazineacetamide, N-(4-aminophenyl)-N,4-dimethyl-

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

Study period:

23 january 2003 - 21 february 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study (OECD 471) and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

BIBF 1120/CDBB 213 BS

IUPAC Name:

BIBF 1120/CDBB 213 BS

Details on test material:

- Name of test material (as cited in study report): BIBF 1120/CDBB 213 BS
- Physical state: weak yellow-white solid substance
- Analytical purity: 99.2 % HPLC
- Purity test date: 2 april 2002
- Lot/batch No.: 8260050
- Expiration date of the lot/batch: 31 january 2003
- Storage condition of test material: at room temperature in the dark (ambient humidity)

Method

Target gene:

S. typhimurium
TA 1537 hisC3076 rfa uvrB - Frameshift
TA 98 hisD3052 rfa uvrB pKM101 Frameshift
TA 100 hisG46 rfa uvrB pKM101 Base substitution
TA 1535 hisG46 rfa uvrB - Base substitution
TA 102 hisG428 rfa + pKM101, pAQ1 Base substitution

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

100, 1000, 2500, 5000, 7500 µg/plate of the test substance

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

mitomycin C

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation

DURATION
- Exposure duration: 2 days

NUMBER OF REPLICATIONS: 3 each per concentration level and positive control and 6 per negative control

Evaluation criteria:

A reproducible, concentration-dependent increase in the number of revertants of at least one tester strain over the vehicle control value and/or outside the historical control range is indicative of genotoxic activity. Since the results were unequivocal, no detailed statistical evaluation was performed.

Statistics:

The assay was considered valid since the following criteria were met:
All tester strains exhibited a characteristic number of spontaneous revertants per plate. The ranges experienced in our laboratory covering about 70 experiments were as follows:
TA 1535: 2-20
TA 1537: 0-15
TA 98: 8-49
TA 100: 28-114
TA 102: 64-239

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Neither precipitation, nor bacteriotoxicity, as been by a reduced background lawn and/or a decrease of absolute revertants, was observed up to the highest concentration.

BIBF 1120/CDBB213BS did not increase the number of revertants after exposure up to the maximum concentration of 7500 µg/plate using the plate incorporation method. Addition of S9 liver fraction from rats pretreated with Aroclor 1254 had no influence on the mutation frequency. The negative were quantitatively confirmed in the repeat experiment using the preincubation method.

Detailed test results are attached as pdf.

Applicant's summary and conclusion

Conclusions:

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

The test substance caused neither base-pair substitutions nor frameshift mutations up to concentrations of 7500 µg/plate when tested in different strains of S. typhimurium in the presence and absence of metabolic activation. Therefore, the test substance can be classified as 'Ames negative'.