1-ethyl-1-methylpyrrolidinium bromide

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

All  positive control groups showed significantly increased mutation frequencies which demonstrates the sensitivity of the test system.

the test substance was not toxic up to 5 mg/plate. all used concentrations could be evaluated.

In none of the tested concentrations and with none of the used strains a statistically significant increase of the mutation frequency was obtained. Metabolic activation did not change these results.

The test substance is therefore non mutagenic in Ames test.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

October-December 1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

under GLP requirenents

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial forward mutation assay

Specific details on test material used for the study:

Cas # 69227-51-6
Batch # 0691
pH: 6-9
Storage: 5°C in the dark
Supplier: Chemson, Polymer-Additive Gesellschaft m.b.H. A-9601 Arnoldstein.
Characterization:
Appearance: Colorless liquid.
pH 6.84
Content 52.2% (estimation of bromide)

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium TA 97

Remarks:

Mutation D6610, rfa,uvrB, pkM101

Additional strain / cell type characteristics:

other: D6610 is a frameshift mutation, rfa leads to reduced lipopolysaccharide barriere in the cell wall . uvrB results in a loss of the DNA excision repair system. pkM101 increases the sensitivity to mutagens as well.

Species / strain / cell type:

S. typhimurium TA 98

Remarks:

Mutation D3052, rfa, uvrB, pkM101

Additional strain / cell type characteristics:

other: D3052 is a frameshift mutation, rfa, uvrB, pkM101 as in TA 97

Species / strain / cell type:

S. typhimurium TA 100

Remarks:

Mutation G46 , rfa, uvrB, pkM101

Additional strain / cell type characteristics:

other:

Remarks:

G46 is a base pair mutation, rfa, uvrB, pkM101 as in TA97

Species / strain / cell type:

S. typhimurium TA 102

Remarks:

Mutation G428, rfa, pkM101

Additional strain / cell type characteristics:

other: G428 is an ochre mutation with sensitivity to hydroperoxides and crosslinking substances, rfa and pkM101 as in TA97

Metabolic activation:

with and without

Metabolic activation system:

microsomal fraction of homogenized livers of rats treated once with 500 mg/kg arcolor 1240

Test concentrations with justification for top dose:

high concentration 1: 5000 µg/plate 3 samples
concentration 2: 1667 µg/plate 3 samples
concentration 3: 555 µg/plate 3 samples
concentration 4: 185 µg/plate 3 samples
low concentration 5: 62 µg/plate 3 samples
control (water): 6 samples
Postive control: Aflatoxine B1, 1µg/plate, for strains TA97 and TA100 at samples with S9-mix.

Vehicle / solvent:

water

Untreated negative controls:

yes

Negative solvent / vehicle controls:

other: water

Positive controls:

yes

Positive control substance:

other: Aflaxotine B1

Details on test system and experimental conditions:

see attached document on test system (1-3)

Rationale for test conditions:

The study was conducted according to OECD 471. according to Ames (1983) mutationRes. 113, 173-215 the strains TA97 and TA 102 are recommended instead of TA1535 and TA1537 and they give better response to the mutagen spectrum. theses are the strain used in this test.

Statistics:

see attached document on test system (1-3)

Key result

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks:

tested only with metabolic activation

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks:

tested without metabolic activation

Remarks on result:

other: not mutagenic

Conclusions:

All positive control groups showed significantly increased mutation frequencies which demonstrates the sensitivity of the test system.
the test substance was not toxic up to 5 mg/plate. all used concentrations could be evaluated.
In none of the tested concentrations and with none of the used strains a statistically significant increase of the mutation frequency was obtained. Metabolic activation did not change these results.
The test substance is therefore non mutagenic in Ames test.

Executive summary:

MEP was tested for mutagenic action with the salmonella thyphimurium reverse mutation assay (Ames test). The study was conducted according to OECD 471.

The test substance was tested at concentrations ranging from 62 µg to 5 mg per plate. with and without external metabolizing system S9 -mix.

The following bacterial strains were used: TA 97, TA 98, TA 100 and TA 102, as well as negative (vehicle) and positive control.

All  positive control groups showed significantly increased mutation frequencies which demonstrates the sensitivity of the test system.

the test substance was not toxic up to 5 mg/plate. all used concentrations could be evaluated.

In none of the tested concentrations and with none of the used strains a statistically significant increase of the mutation frequency was obtained. Metabolic activation did not change these results.

The test substance is therefore non mutagenic in Ames test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

A slight cytotoxic effect of the test substance was noted at all sampling times in animals of both sexes.

The rate of micronucleated polychromatic erythrocytes, which is the target of the study, was increased significantly by the test substance at all sampling times, compared to the negative control group and to historical negative control data.

While no differences between the sexes were noted considering mutagenicity, cytotoxicity was more pronounced in females.

The test substance does produce micronuclei in polychromatic erythrocytes at a dose of 350 mg/kg body weight at sampling time of 24, 48 and 72 hr p.a

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May-October 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

under GLP guidelines

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

yes (incl. QA statement)

Type of assay:

other: Mammalian Erytrocyte Micronucleous Test

Specific details on test material used for the study:

Cas # 69227-51-6
Batch # 0691
pH: 6-9
Storage: 5°C in the dark
Supplier: Chemson, Polymer-Additive Gesellschaft m.b.H. A-9601 Arnoldstein.
Characterization:
Appearance: Colorless liquid.
pH 6.84
Content 52.2% (estimation of bromide)

Species:

mouse

Strain:

NMRI

Remarks:

BR

Details on species / strain selection:

Justification for the selection of the species: Recommended by the guideline

Sex:

male/female

Details on test animals or test system and environmental conditions:

Supplier : Charles River WIGA GmbH (D-8741 Sulzfeld 1)
number in main study 25 males and 25 females
Age: approx. 10 weeks
Body weight (average; gr): males; NC 31.2, PC 30.5, A1 31.1, B1 30.7, C1 30.6
Body weight (average; gr): females; NC 23.5, PC 22.3, A1 23, B1 22.7, C1 22.6
Hygiene: improved conventional conditions
Room number: PHA-18
Room temperature: average of 24°C
Relative humidity: average of 50%
air exchange: 12 per hr
Light: artificial light from 6 am to 6 pm
Cages: Makrolon type III females, type II males
five animals per cage (females), single caging (males)
Bedding material: aspen wood chips, autoclaved
Feed: Altromin 1314 ff, gamma irradiated with 10kGy 60Co, ad libitum
Exception: feed was withdrawn the evening before application and was offered again about one hr after application. Random samples of the food are analysed for contaminants by Altromin, D-4937 Lage.
Water: tap water, from makrolon bottles with stainless steel canules, ad libitum
Identification: labelling with felt-tipped pen on the tail and the cage.
Acclimatization: 10 days.

Route of administration:

oral: gavage

Vehicle:

Distilled water was used for the solution of the test substance and for Negative Control (NC) (dose volume 10 ml/kg body weight).
The test substance was diluted with distilled water and was applied once at a dose of 350 mg/kg body weight by stomach intrubation.
Based on two preliminary range finding study, a dose of 350 mg of test substance per kg body weight was chosen for main study
The negative as well as positive control group was tested as well.

Details on exposure:

The test substance was diluted with distilled water and was applied once

Duration of treatment / exposure:

The animals were killed 24, 48 and 72 hr post application.
One negative control group (distilled water, killed 48 hr p.a) and one positive control (cyclophosphamide, killed 24 hr p.a) were included in the study.

Frequency of treatment:

Applied once

Dose / conc.:

350 mg/kg bw/day (nominal)

Remarks:

test material

Dose / conc.:

10 other: ml/kg b.w

Remarks:

distilled water (negative control)

Dose / conc.:

10 other: ml/kg b.w

Remarks:

cyclophosphamide (positive control)

No. of animals per sex per dose:

total of 25 males and 25 females (3 groups each)
5 males and 5 females for positive and negative control

Control animals:

yes

Positive control(s):

Cyclophosphamide (Positive Control, PC) was dissolved in distilled water (dose volume 10 ml/kg body weight)

Tissues and cell types examined:

Bone marrow

Details of tissue and slide preparation:

see attached document on methods

Statistics:

see attached document on methods

Key result

Sex:

male/female

Genotoxicity:

positive

Toxicity:

yes

Remarks:

slight cytotoxic effects, more pronounced in females

Vehicle controls validity:

valid

Remarks:

Negative control

Negative controls validity:

valid

Remarks:

No effect

Positive controls validity:

valid

Remarks:

micronuclei induction as a result of damage or damage to mitotic apparatus in vivo. have cytotoxic properties. amount of polychromatic erythrocyes raised and nucleated cells lowered.

Additional information on results:

see attached document on results

Conclusions:

A slight cytotoxic effect of the test substance was noted at all sampling times in animals of both sexes.
The rate of micronucleated polychromatic erythrocytes, which is the target of the study, was increased significantly by the test substance at all sampling times, compared to the negative control group and to historical negative control data.
While no differences between the sexes were noted considering mutagenicity, cytotoxicity was more pronounced in females.

The test substance does produce micronuclei in polychromatic erythrocytes at a dose of 350 mg/kg body weight at sampling time of 24, 48 and 72 hr p.a

Executive summary:

The test substance N-Methyl-Ethyl-Pyrollidinium-Bromid (MEP) was administered to 3 groups of 5 males and 5 female mice each. The test substance, dilluted with distilled water, was applied once at the dose of 350 mg/kg b.w.

The animals were killed 24, 48 and 72 hr p.a. Preperations of bone marrow cells were conducted according to OECD 474.

Negative and positive controls were included in the study as well.

Results: A slight cytotoxic effect of the test substance was noted at all sampling times in animals of both sexes.

The rate of micronucleated polychromatic erythrocytes, which is the target of the study, was increased significantly by the test substance at all sampling times, compared to the negative control group and to historical negative control data.

While no differences between the sexes were noted considering mutagenicity, cytotoxicity was more pronounced in females.

The test substance does produce micronuclei in polychromatic erythrocytes at a dose of 350 mg/kg body weight at sampling time of 24, 48 and 72 hr p.a

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (positive)