1-ethyl-1-methylpyrrolidinium bromide

Endpoint summary

Description of key information

None of the parameters determined in this study was clearly or significantly changed due to a toxic action of the test material.

The lower body weight in high dosed males cannot be given relevance due to lack of statistical significance. A changed stain of the bedding material does not represent a toxic change.

No alternations which were related to the action of the test substance were noted in dosed animals.

Based on this 150 mg of the test substance per kg body weight can be defined as No-Adverse-Effect-Level.

Results of the corresponding dose range finding study demonstrate, that toxic doses are only slightly above the NOEL, determined in the present study.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: oral, other

Remarks:

Sub - acute

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Marc-July 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

Chemical nature: 50% aqueous solution of the test substance
CAS number: 69227-51-6
Batch number: 0691
Storage: RT in the dark
Label on container: N-Ethyl, N-Methyl-Pyrrolidinium-Bromid (MEP)
Manufacturer: Chemson, Polymer-Additive Gesellschaft m.b.H A-9601 Arnoldstein.
colorless liquid

Species:

rat

Strain:

Wistar

Details on species / strain selection:

Source: Charles River WIGA GmbH., D-8741 Sulzfeld
Number 20 males and 20 females
Body weight:
Average Control group
Average K1 males: 154 (-6 day) - 422 (27 day) (gr)
Average K1 females: 143 (-6day) - 262 (27 day) (gr)
Average group A1 males (low dose) 155 (-6 day) - 410 (27 day) (gr)
Average group A1 females (low dose) 143 (-6 day) - 264 (27 day) (gr)
Average group B1 males (mid dose) 155 (-6 day) - 429 (27 day) (gr)
Average group B1 females (mid dose) 143 (-6 day) - 260 (27 day) (gr)
Average group C1 males (high dose) 155 (-6 day) - 403 (27 day) (gr)
Average group C1 females (highd dose) 143 (-6 day) - 270 (27 day) (gr)
Age at first administration: approx. 6 weeks

Sex:

male/female

Details on test animals or test system and environmental conditions:

Hygiene: maintenance behind a hygienic barrier
Roon number TOX 16
Room temprature: average of 21°C, deviation of +/- 0.3°C
Relative humidity: average of 60%, range between 52% and 65%.
Air exchange: 12 per hr.
Light: artificial light from 6 am to 6 pm
Cages: single caging in Makrolon cages type III. Wire mesh lids. Sansitization of cages once a week.
Bedding material: aspen wood chips, type "4 HV"
Feed: Altromin 1314 ff, gamma irradiated with 10 kGy 60 Co, ad libitum.
Exception: feed was withdrawn on the days before blood sampling at about 5 pm and offered again immediately after blood sampling.
Water: Deionized water, UV irradiated and acidified with HCl to pH 3, from an automatical watering system, ad libitum.
Acclimatization: 12 days.

Route of administration:

oral: gavage

Details on route of administration:

Oral administration was performed by gavage once a day on 5 days per week in the morning. On the day with blood sampling, when feed had been withdrawn over night, test substance dilution was administered after giving the animals again access to the feed for at least 2 hr.
individual dose volume was calculated using the last determined body weight.

Vehicle:

water

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Feed: Random saples of the feed are analysed for contaminants by Altromin GmbH D-4937 (supplier) and one saple was additionaly analysed for contaminants by Landwirtschaftliche Untersuchungs und Forschungsanstalt Kiel.
Acceptable limits of contaminants were derived from the Futtermittelverordnung (animal feed regulation) of the Federal Republic of Germany.
Water: Random samples of water are analysed by the Bundesanstalt fur Lebensmitteluntersuchung und forschung, A-1090 Vienna, to assure that the water fulfills the requirements for drinking water for humans (except for pH).

Duration of treatment / exposure:

4 weeks (28 days)

Frequency of treatment:

Test substance was administered orally by gavage to three groups of 5 males and 5 females each, once a day on 5 days per week for 4 weeks. An equally sized control group received the carrier of the test substance.

Dose / conc.:

15 mg/kg bw/day (nominal)

Remarks:

group A1 low dose; the test substance was applied freshly diluted with disstiled water. The dose volume was 10 ml per kg body weight.

Dose / conc.:

47 mg/kg bw/day (nominal)

Remarks:

group B1 mid dose, the test substance was applied freshly diluted with disstiled water. The dose volume was 10 ml per kg body weight.

Dose / conc.:

150 mg/kg bw/day (nominal)

Remarks:

group C1 high dose, the test substance was applied freshly diluted with disstiled water. The dose volume was 10 ml per kg body weight.

No. of animals per sex per dose:

10 (5 males and 5 females per dose)

Control animals:

yes, concurrent vehicle

Positive control:

no

Observations and examinations performed and frequency:

The following investigations were performed:
Observations in life (clinical signs; behavioural changes, at least once a day)
Body weight (-6 and then once a week on days 1 through 27)
Feed consumption (-5 - 1; and then once a week on day 1 through 27)
Haematology (day 28)
Clinical chemistry (day 28)
Groos pathological examination (animals killed at day 29 by CO2)
Organ weight determination (animals killed at day 29 by CO2)
Histopatological examination (animals killed at day 29 by CO2)

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see attached file)

HISTOPATHOLOGY: Yes (see attached file)

Statistics:

Analysis of variance followed by the Schffe test. For aritmetic means and standard deviations, when more than two groups are compared.
H-test of Kruskal and Wallis followed by the test of Nemeny. Instead of analysis of variance, if requirements for this test are not met or for counted events.
T-test: for aritmetic means and Standard deviations when only two groups are compared.
Chi-square Test: for counted events
Fishers Exact test: for counted events, if requirements of Chi-Square-Test are not fulfilled.
U test: For counted events, where a grade of severity is available.
Results were analysed seperately for males and females
p=0.05 was chosen in each test

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

group K1 (control): all animals were normal
group A1 (low dose): unspecific signs; alopecia and dermal erosions in one female
group B1 (mid dose): unspecific signs; alopecia and dermal erosions in two males
group C1 (high dose): Besides unspecific signs; alopecia and dermal erosions, soft faeces was found in one male for one day. A dark staining of bedding material was noted on day 9 through day 18 in all animals.
staining of bedding material noted above remains without interpretation. The stain itself gave no indication for a possible lesion, like haematuria or Jaundice. Therefore it is not considered to represent a toxic change.

Mortality:

no mortality observed

Description (incidence):

All animals survived until termination of the study.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Body weight was not significantly lower on days 14 throught 27 in high dosed males, as compared to control

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

In all dosed animals of both sexes feed consumption was comperable to the control during the whole study.

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

group A1: no significant differences compare to control K1
group B1: MCH (mean corpuscular haemoglobin); males, increased; MCHC (MCH concentration) males, increased
group C1: no significant differences compare to control K1

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

group A1: no significant differences compare to control K1
group B1: significant decrease of serum potassium concentration was noted in males. There is a similar trend in females, but the decrease didnt have statistical significance.
group C1: no significant differences compare to control K1

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

group A1 and group B1: no significant differences in absolute and relative organ weights were noted as compared to the controls
group C1: no significant differences in absolute and relative organ weights were noted as compared to the controls. higher weights of spleens, noted in females, lacked to gain statistical significance.

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

see above

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

group K1: alternation in the liver (i.e. panlobular lymphohistiocytic foci) and of the heart (i.e. foca chronic mycocarditis) respectively was observed.
group A1: as there were no indications of test substance related alternations noted in high dosed animals, no histopathological examinations were performed.
group B1: mandibular lymph nodes of one male and spleen of one female were analysed to define unclear necrotic findings. Both organs showed normal histophatology.
as there were no indications of test substance related alternations noted in high dosed animals, no further histopathological examinations were performed
group C1: alternations of the kidneys, heart and liver respectively were observed. Mandibular lymph nodes and the thymus were taken from one female respectivelly, one male to define unclear necropsy findings. Lymph nodes were found to be normal histopathogically. A focal haemorrhage was noted in single thymus examined.
Those findings are known to occur spontaneously in animals of this strain in a comparable incidence. Therefore these alterations are not related to the action of the test substance.

Histopathological findings: neoplastic:

not examined

Other effects:

effects observed, non-treatment-related

Description (incidence and severity):

Necropsy:
group K1: all animals were normal at necropsy
group A1: all animals were normal at necropsy
group B1: large mandibular lymph nodes were noted in one male, a large spleen in one female
group C1: large mandibular lymph nodes were noted in one female, thymus petechiae in one male
All findings are considered to be spontaneous changes, all were of single occurence.

Key result

Dose descriptor:

NOEL

Effect level:

150 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: The following investigations were performed:Observations in life, Body weight, Feed consumption, Haematology,Clinical chemistry, Groos pathological examination, Organ weight determination and Histopatological examination.

Remarks on result:

not determinable due to absence of adverse toxic effects

Critical effects observed:

no

Conclusions:

None of the parameters determined in this study was clearly or significantly changed due to a toxic action of the test material.
The lower body weight in high dosed males cannot be given relevance due to lack of statistical significance. A changed stain of the bedding material does not represent a toxic change.
No alternations which were related to the action of the test substance were noted in dosed animals.
Based on this 150 mg of the test substance per kg body weight can be defined as No-Adverse-Effect-Level.
Results of the corresponding dose range finding study demonstrate, that toxic doses are only slightly above the NOEL, determined in the present study.

Executive summary:

This 28 day toxicity study was conducted according to OECD 407 and EC gideline B7.

The test stbstance N-Methyl-N- Ethyl-Pyrollidinium-Bromid (MEP) was administered orally to 3 groups of rats:

Low dose: 15 mg/kg b.w; Mid dose: 47 mg/kg b.w and High dose 150 mg/kg b.w (5 males/females each), omce a day on 5 days per week for 4 weeks. An equaly control group received the carrier of the test substace (distilled water). The dose volume as 10 ml per kg b.w.

The following investigations were performed:Observations in life, Body weight, Feed consumption, Haematology,Clinical chemistry,

Groos pathological examination, Organ weight determination and Histopatological examination.

None of the parameters determined in this study was clearly or significantly changed due to a toxic action of the test material.

The lower body weight in high dosed males cannot be given relevance due to lack of statistical significance. A changed stain of the bedding material does not represent a toxic change.

No alternations which were related to the action of the test substance were noted in dosed animals.

Based on this 150 mg of the test substance per kg body weight can be defined as No-Adverse-Effect-Level.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

150 mg/kg bw/day

Study duration:

subacute

Species:

rat