(3-chloro-2-hydroxypropyl)trimethylammonium chloride

Genetic toxicity: in vivo

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

1983

Test guideline 2

Qualifier:

according to guideline

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

Test material information

Constituent 1

Reference substance name:

Automatically generated during migration to IUCLID 6, no data available

IUPAC Name:

Automatically generated during migration to IUCLID 6, no data available

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 or test system 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.