Registration Dossier

Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990-10-15 to 1991-06-24
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
1983
Qualifier:
according to
Guideline:
EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
1984
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
Quab 188, approx. 60 % aqueous solution of a 99.92 % pure solid

Test animals

Species:
mouse
Strain:
other: BOR: NMRI (SPFHan)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Winkelmann, Vrsuchxtierzucht GmbH & Co.KG, D-4799 Borche, Germany
- Age at study initiation: 5 weeks
- Weight at study initiation: males 29-35 g; females 23-31g
- Assigned to test groups randomly: yes
- Fasting period before study: no
- Housing: Macrolon cages type II; 1 per cage
- Diet: ad libitum
- Water: ad libitum
- Acclimation period:


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 55
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle:
- Concentration of test material in vehicle:
Duration of treatment / exposure:
single administration; observation time: 72 h
Frequency of treatment:
single treatment
Post exposure period:
72 hours
Doses / concentrations
Remarks:
Doses / Concentrations:
147 mg/kg body weight
Basis:

No. of animals per sex per dose:
6
Control animals:
other: physiological saline
Positive control(s):
- cyclophosphamide
- Justification for choice of positive control(s): none given, standard control substance
- Route of administration: oral gavage
- Doses / concentrations: 51.1 mg/kg

Examinations

Tissues and cell types examined:
Bone marrow
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: Based on preliminary toxicity evaluation

DETAILS OF SLIDE PREPARATION: air dried slides - no other details

METHOD OF ANALYSIS: 1000 PCEs scored for micronuclei under microscope (650-1000 x magnification); NCE/PCE calculated on data from 1000 erythrocytes
Evaluation criteria:
A test material is considered positive if it produces a statistically significant positive response in all 3 test points.
Statistics:
Poisson test applied

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Remarks:
5 deaths
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 100-215 mg/kg
- Clinical signs of toxicity in test animals: severe symptoms of toxicity at all dosese, deaths occurred at highest dose
- Evidence of cytotoxicity in tissue analyzed: no abnormalities detected at necroscopy of treated animals
- Rationale for exposure: based on MTD

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): negative overall
- Ratio of PCE/NCE (for Micronucleus assay): not reduced in test material group animals when compared with corresponding negative control, except for one male at 48 hour sampling time
- Appropriateness of dose levels and route: both appropriate
- Statistical evaluation: showed slightlly statistically significant increase in micronucleated PCEs (p= 0.04) when data for both sexes combined. This was not considered biologically significant as it was due to the lower than heistorical value negative control, especially in males.

Any other information on results incl. tables

Table 1 Results of in vivo micronucleus test (5 animals/sex, 1000 cells evaluated per animal) 24 hour sampling time

Sampling time

Treatment

Male/female

PCE with MN

Ratio PCE/NCE*

24 hours

Negative Control

male

1.6 ± 1.14

1.71-2.45

female

1.4 ± 0.55

1.67-1.97

Test substance

male

1.6 ± 1.14

1.41-3.31

female

1.2 ± 1.07

0.96-1.82

Positive control

male

30.8 ± 13.66

1.70-2.24

female

27.6 ± 4.45

0.90-1.82

 

Table 2 Results of in vivo micronucleus test (5 animals/sex, 1000 cells evaluated per animal) 48 hour sampling time

Sampling time

Treatment

Male/female

PCE with MN

Ratio PCE/NCE*

48 hours

Negative Control

male

0.6 ± 0.89

1.20-2.09

female

1.6 ± 0.55

1.42-2.13

Test substance

male

1.6 ± 1.14

0.69-2.10

female

2.8 ± 1.30

1.58-2.88

Positive control

male

18.8 ± 3.77

0.32-0.62

female

5.8 ± 2.77

0.75-1.19

Table 3 Results of in vivo micronucleus test (5 animals/sex, 1000 cells evaluated per animal) 72 hour sampling time

Sampling time

Treatment

Male/female

PCE with MN

Ratio PCE/NCE*

72 hours

Negative Control

male

0.4 ± 0.55

2.55-4.05

female

0.5 ±0.55

1.78-3.47

Test substance

male

1.6 ±0.55

1.69-3.93

female

0.5 ± 0.84

1.86-3.22

Positive control

male

7.0 ± 1.73

0.16-0.46

 

3.8 ± 1.3

0.34-1.31

 

 

 

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
3-chloro-2-hydroxypropyltrimethylammonium chloride has been tested in a valid study according to OECD 474 and under GLP. No biologically or statistically significant increase in the number of PCEs with micronuclei was observed at 24, 48 or 72 hours after administration by ip injection. It is noted that the test substance did not cause a reduction in the PCE/NCE ratio.