2-piperazin-1-ylethanol

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

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:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

May 1983/ Revised Draft, March 1996

Deviations:

yes

Remarks:

(2-aminoanthracene was the only used positive control in experiments with metabolic activation)

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)

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

December 1992

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-piperazin-1-ylethanol

EC Number:

203-142-3

EC Name:

2-piperazin-1-ylethanol

Cas Number:

103-76-4

Molecular formula:

C6H14N2O

IUPAC Name:

2-piperazin-1-ylethanol

Test material form:

liquid

Specific details on test material used for the study:

- Batch No.: 25-7626
- storage: room temperature

Method

Target gene:

his and trp operon

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

other: defective excision repair system

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Additional strain / cell type characteristics:

not specified

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
- Justification for choice of solvent/vehicle: good solubility

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

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 results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A slight decrease of revertants was occasionally observed from 2500 µg/plate onwards.

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A slight decrease of revertants was occasionally observed from 2500 µg/plate onwards.

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A slight decrease of revertants was occasionally observed from 2500 µg/plate onwards.

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

A slight decrease of revertants was occasionally observed from 2500 µg/plate onwards.

Vehicle controls validity:

valid

Positive controls validity:

valid

Test results 5

Key result

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

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No Test substance precipitation was found.

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±2	21±2	140±17	151±14	14±1	16±2	32±5	39±4	32±2	42±5
20	20±1	21±1	154±7	151±16	11±2	13±2	33±4	41±7	33±3	37±3
100	19±1	20±2	145±9	147±12	10±3	10±1	32±3	43±7	31±1	39±3
500	22±2	20±2	147±19	139±11	10±0	10±1	34±7	29±2	29±1	38±2
2500	21±1	13±2	127±6	136±7	11±1	9±1	28±3	27±4	27±5	35±4
5000	20±1	11±1	140±6	140±6	8±2	11±3	31±3	22±5	27±3	33±3
2-AA	-	223±13	-	1431±6	-	118±4	-	1005±9	-	266±37
MNNG	1119±105	-	1025±56	-	-	-	-	-	-	-
AAC	-	-	-	-	307±4	-	-	-	-	-
NPD	-	-	-	-	-	-	1011±7	-	-	-
ENNG	-	-	-	-	-	-	-	-	512±13	-

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	21±3	22±4	140±6	160±1	10±1	12±0	31±1	45±1	36±1	50 ± 2
20	19±3	18±3	144±11	158±3	11±2	11±1	29±3	45±2	34±2	52 ± 6
100	21±2	20±4	142±7	153±8	10±1	9±2	24±4	40±2	31±3	47 ± 2
500	21±2	17±2	160±11	155±5	10±2	12±2	26±1	43±3	34±4	44 ± 4
2500	17±2	16±1	169±10	162±8	9±2	9±3	29±5	38±2	31±5	44 ± 6
5000	16±2	10±2	158±16	161±1	8±2	9±2	26±2	45±2	32±5	42 ± 6
2-AA	-	149±11	-	1484±45	-	174±23	-	1226±163	-	223 ±25
MNNG	1644±231	-	2034±58	-	-	-	-	-	-	-
AAC	-	-	-	-	712±11	-	-	-	-	-
NPD	-	-	-	-	-	-	1281±64	-	-	-
ENNG	-	-	-	-	-	-	-	-	670±44	-

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:

neagtive Ames test

Executive summary:

A study according to OECD TG 471 was conducted to assess the in vitro gene mutation potential of the test item in different bacteria strains (TA 1537, TA 1535, TA 98, TA 100 and E. coli WP uvrA). In the preincubation method bacteria were preincubated for 20 min with the test item using concentrations of 20, 100, 500, 2500 and 5000 µg/plate. Water was used as solvent control. Afterwards, bacterial were plates and exposed for 48 to 72 h. In the standard plate test the exposure duration was 48 to 72 hours as well. Incubation was done in the presence and in the absence of a metabolizing system (S9-Mix from Aroclor-1254 treated male rats). With regard to cytotoxicity, a slight decrease of revertants was occasionally observed from 2500 µg/plate onwards. No genotoxicity was observed neither in presence nor absence of S9-mix. The positive controls induced a significant increase in mutation frequency.