1-ethylpiperazine

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:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

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)

Test guideline 2

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

Test material information

Constituent 1

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)

Controls

Controls 1

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

Controls 2

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