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Toxicological information

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
(2-aminoanthracene was used as sole indicator of the efficacy of the S-9 mix)
Qualifier:
according to guideline
Guideline:
OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)
Version / remarks:
Date of deletion: 21 July 1997 (Method merged with TG 471)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1-ethylpiperazine
EC Number:
226-166-6
EC Name:
1-ethylpiperazine
Cas Number:
5308-25-8
Molecular formula:
C6H14N2
IUPAC Name:
1-ethylpiperazine
Details on test material:
- Name of test material (as cited in study report): N-Ethylpiperazin
- Physical state: Colorless to light yellow liquid
- Analytical purity: 99.6%
- Batch No.: Abl. Nr. 85-6636
- Storage condition of test material: Room temperature

Method

Target gene:
his-, trp-operon
Species / strain
Species / strain / cell type:
other: S. typhimurium TA 100, TA 1535, TA 1537, TA 98 and E. coli WP2 uvrA
Metabolic activation:
with and without
Metabolic activation system:
Aroclor induced rat liver S-9 mix
Test concentrations with justification for top dose:
20, 100, 500, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Water (DMSO was used as vehicle for the positive controls)
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene (2.5 µg/plate for TA100, TA98, TA1537 and TA1535, 60 µg/plate for E.coli)
Remarks:
with metabolic activation
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: N-methyl-N-nitro-N-nitrosoguanidine (5 µg/plate for TA100, TA1535), N-ethyl-N-nitro-N-nitrosoguanidine (10 µg/plate for E.coli), 9-aminoacridine (100 µg for TA1537), 4-nitro-o-phenylendiamine (10 µg/plate for TA98)
Remarks:
without metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: standard plate test and preincubation test both with and without metabolic activation

DURATION
- Preincubation period: at 37°C for the duration of 20 minutes
- Exposure duration: at 37°C for 48 - 72 hours in the dark (for standard plate test and preincubation test)
Evaluation criteria:
In general, a substance to be characterized as positive in the bacterial tests has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA 100, TA 1535, TA 1537, TA 98 and E. coli WP2 uvrA
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Remarks:
(but tested up to limit dose)
Vehicle controls validity:
valid
Positive controls validity:
valid
Additional information on results:
No precipitation of the test substance was found.
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Standard 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±1

20±1

135±5

131±6

12±3

10±2

31±3

39±7

33±2

38±3

 20

17±2

19±2

127±7

116±7

9±4

15±4

27±2

34±5

32±6

34±3

 100

16±2

16±1

127±14

127±17

11±1

9±3

28±1

37±3

31±4

31±5

 500

15±1

15±1

108±17

120±22

7±2

11±3

24±3

34±4

32±4

34±4

 2500

13±2

15±3

98±17

128±10

9±3

9±3

19±6

30±4

38±10

34±2

 5000

13±1

14±0

99±12

102±16

11±2

12±3

19±2

28±2

31±1

30±2

 2-AA

 -

 159±30

 -

1298±60 

 -

 204±6

 -

 1211±32

 -

 233±9

 MNNG

 1012±26

 -

 1483±63

 -

 -

 -

 -

 -

- 

 -

 AAC

 -

 -

 -

 -

 475±51

 -

 -

 -

 -

 -

 NPD

 -

 -

 -

 -

 -

 -

 956±42

 -

 -

 -

 ENNG

 -

 -

 -

-

 -

 -

 -

 -

 517±14

 -

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±1

21±2

114±4

123±5

10±1

11±1

30±3

43±4

30±3

48±6

 20

19±2

18±2

121±22

149±12

10±3

12±2

27±2

41±1

31±1

44±4

 100

18±2

17±1

146±13

159±15

10±3

11±1

29±5

38±4

31±5

47±10

 500

17±1

15±4

137±19

152±14

10±0

12±1

28±2

29±4

32±3

47±7

 2500

18±2

16±1

150±2

158±7

10±1

10±2

29±3

31±2

27±2

42±3

 5000

16±2

10±1

109±8

158±6

10±1

9±2

19±4

29±3

20±3

48±3

 2-AA

 -

 136±11

 -

1137±90 

 -

 124±11

 -

 1046±64

 -

 183±12

 MNNG

 1295±50

 -

 1068±14

 -

 -

 -

 -

 -

- 

 -

 AAC

 -

 -

 -

 -

 520±70

 -

 -

 -

 -

 -

 NPD

 -

 -

 -

 -

 -

 -

 995±41

 -

 -

 -

 ENNG

 -

 -

 -

-

 -

 -

 -

 -

 1129±122

 -

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

Applicant's summary and conclusion

Conclusions:
negative Ames test