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Genetic toxicity: in vitro

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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:
11. May 1997 - 01. Jul 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline Study (OECD 471/472)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
; OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): N-Methylpiperazin
- Physical state: colorless liquid
- Analytical purity: 99.8 %
- Lot/batch No.: 25-0388
- Storage condition of test material: Room temperature (N2 conditions).

Method

Target gene:
Histidin operon
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9-mix from Aroclor 1254 treated male Sprague-Dawley rats.
Test concentrations with justification for top dose:
0, 20, 100, 500, 2500 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controls
Untreated negative controls:
yes
Remarks:
sterility control
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: with S-9 mix: all strains 2-aminoanthracene; without S-9 mix: strains TA100, TA1535: N-methyl-N'-nitro-N-nitrosoguanidine; TA98: 4-nitro-o-phenylendiamine; TA1537: 9-aminoacridine; E.coli WP2 uvrA: N-ethyl-N'-nitro-N-nitrosoguanidine.
Details on test system and experimental conditions:
METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 - 72 h


NUMBER OF REPLICATIONS: 3

METHOD OF APPLICATION: standard plate test

DURATION
- Exposure duration: 48 - 72 h


NUMBER OF REPLICATIONS: 3

Evaluation criteria:
The test chemical is considered positive in this assay if the following criteria are met: A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.
A test substance is generally considered non-mutagenic in this test if: The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: Complete solubility of test substance in aqua dest.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Standart plate test:

 Dose (µg/plate)  TA1535     TA100     TA1537     TA98        E. coli WP2 uvrA            
   -S9  +S9  -S9  +S9  -S9  +S9 -S9  +S9 -S9  +S9             
 0 20±2 20±1 153±17 158±9 13±4 12±3 31±3 45±3 37±7 36±5            
20  18±1 17±1 147±12 161±9 13±2 12±2 34±3 42±1 31±1 26±3            
 100 17±1 16±1 163±5 157±12 11±1 16±2 32±2 37±2 34±4 30±2            
 500 14±3 20±1 153±7 157±16 11±2 14±2 34±3 38±6 34±3 32±3            
 2500 15±2 17±2 152±14 156±5 13±4 14±2 32±3 42±3 28±3 31±3            
 5000 14±2 17±3 154±6 163±5 11±2 12±1 33±2 40±2 29±1 34±7            
 2-AA - 149±7 - 1474±62  - 179±2 1237±32  - 317±37            
 MNNG  955±36 - 858±42 - -  - -  - - -            
 AAC - - - - 750±6 - - - - -            
 NPD  - - - - - - 1104±77 -            
 ENNG  - - - - - - - - 1185±189 -            

Mean ± SD

Preincubation-test:

 Dose (µg/plate)  TA1535     TA100     TA1537     TA98     E. coli WP2 uvrA                 
   -S9  +S9  -S9  +S9  -S9  +S9 -S9  +S9  -S9 +S9               
 0 20±2 20±2 129±2 124±19 11±2  11±2 30±1 41±3 29±1 35±3              
20  21±3 17±1 146±16 151±21 8±2  10±1 26±7 39±6 25±3 29±4              
 100 17±1 17±2 143±11 195±18 10±1  10±1 25±3 41±6 25±6 30±4              
 500 17±3 18±1 150±20 153±7 9±1  9±1 21±1 30±10 26±3 32±3              
 2500 17±1 16±2 158±13 176±25 8±2  9±1 23±4 41±1 24±2 28±3              
 5000 12±1 14±3 130±4 173±25 7±2  7±2 20±1 35±6 19±8 33±3              
 2-AA  -  154±22 - 805±107 -  148±10 - 1173±41 - 242±8              
 MNNG 981±45 - 1232±70 - 971±34 - - - -  -              
 AAC  - - - - - - -  -              
 NPD  - - - - -  - 1228±98 -  -              
ENNG - - - - - - - - 875±45 -              

Mean ± SD

2-AA: 2-aminoanthracene;

MNNG; N-methyl-N-nitro-N-nitrosoguanidine

ENNG; N-ethyl-N-nitro-N-nitrosoguanidine

NPD: 4-nitro-o-phenylendiamine

AAC: 9-aminoacridine chloride monohydrate

Under the conditions tested Diethylaminopropylamin is non mutagenic in the bacterial AMES-test with or without metabolic activation.

The positive control gave the expected values.

Applicant's summary and conclusion