1-ethyl-1-methylpyrrolidinium bromide

Genetic toxicity: in vivo

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

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

Test material

Test material information

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)

Test animals

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Exception: feed was withdrawn the evening before application and was offered again about one hr after application.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.

Administration / exposure

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

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

350 mg/kg bw/day (nominal)

Remarks:

test material

Doses / concentrations 2

Dose / conc.:

10 other: ml/kg b.w

Remarks:

distilled water (negative control)

Doses / concentrations 3

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)

Examinations

Tissues and cell types examined:

Bone marrow

Details of tissue and slide preparation:

see attached document on methods

Statistics:

see attached document on methods

Results and discussion

Test results

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

Applicant's summary and conclusion

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