Quaternary ammonium compounds, coco alkyltrimethyl, chlorides

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

micronucleus assay

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 15 February, 1983 to 18 March, 1983

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

not specified

Remarks:

pre-GLP

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Quaternary ammonium compounds, coco alkyltrimethyl, chlorides

EC Number:

263-038-9

EC Name:

Quaternary ammonium compounds, coco alkyltrimethyl, chlorides

Cas Number:

61789-18-2

Molecular formula:

C(n+3) H(2n+10) N Cl (n=8-18)

IUPAC Name:

Quaternary ammonium compounds, cocoalkyl trimethyl, chloride

Constituent 2

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

water

Test material form:

liquid

Details on test material:

- Chemical name: Quaternary ammonium compounds, coco alkyltrimethyl, chlorides
- EC number: 263-038-9

To the best of knowledge, the sample used is representative to the boundary composition of the substance registered

Test animals

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River U.K. Limited, Margate, Kent
- Weight at study initiation: 18-21 g
- Assigned to test groups randomly: Yes
- Housing: Plastic disposable cage
- Diet: Spratt's Laboratory Diet number 1, ad libitum
- Water: Tap water, ad libitum
- Acclimation period: 10 d

ENVIRONMENTAL CONDITIONS
- Air changes (per hr): 30/h
- Photoperiod (hrs dark / hrs light): 12 h/12 h

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

1% methyl cellulose

Details on exposure:

Dilution of the original solution were made in aqueous 1% methyl cellulose to give the desired concentration of the test substance. The mice were starved overnight prior to dosing. All animals in all groups were dosed with the standard volume of 2 mL/10 g bw. The test substance and vehicle control were dosed by oral gavage. The positive control was dosed by intraperitoneal injection.

Duration of treatment / exposure:

Single treatment

Frequency of treatment:

Single

Doses / concentrations

Remarks:

Doses / Concentrations:
468 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

45 males and 45 females

Control animals:

yes, concurrent vehicle

Positive control(s):

Mitomycin C was used. It was prepared as a solution in sterile 0.9% saline at a concentration of 0.4 mg/mL.

Examinations

Tissues and cell types examined:

The femurs were cleared of tissue and one epiphysis removed from each bone.

Details of tissue and slide preparation:

A direct bone marrow smear was made on to a slide containing a drop of calf-serum. One smear was made from each femur. The prepared smears were air-dried and fixed in methanol. After fixation, the smears were air-dried and stained using Giemsa's technique. After rinsing in buffered distilled water, the slides were air-dried and mounted with coverslips using DPX. The stained smears were examined by light microscopy to determine the incidence of micronucleated cells/1,000 polychromatic erythrocytes/animal. The ratio of polychromatic to nonchromatic erythrocyte (NCEs) for each animal was assessed by examination of at least 1,000 erythrocyte.

Evaluation criteria:

Reproducible and significant increase in the number of micronucleated NCEs in the test group over that of the control group.

Statistics:

Non-parametric methods by Hollander M and Wolfe DA.

Results and discussion

Test results

Test results 1

Key result

Sex:

male

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: not clastogenic

Test results 2

Key result

Sex:

female

Genotoxicity:

negative

Vehicle controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: not clastogenic

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
Probit analysis yielded an estimated LD10/3 of 468 mg/kg bw (LD50/3 was 884 mg/kg bw). A dose level of 468 mg/kg bw was chosen for the micronucleus test.

RESULTS OF DEFINITIVE STUDY- Induction of micronuclei (for Micronucleus assay): No significant difference as compared to controls.
- Ratio of PCE/NCE (for Micronucleus assay): The ratio of PCE to total erythrocytes remained essentially unaffected by the test substance
- Appropriateness of dose levels and route: Yes
- Statistical evaluation: Yes

Any other information on results incl. tables

No mortalities was seen in animals. Clinical signs included pilo-erection, hunched posture, lethargy and ptosis.

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the test substance was found to show no evidence of clastogenic potential in the bone marrow cells of mice.

Executive summary:

A study was conducted to determine the clastogenic potential of test substance, Coco TMAC (33% active in water) in an in vivo mouse bone marrow micronucleus test in mice, according to OECD 474 and EU Method B.12 Guidelines. Based on the results of a dose range finding assay, a dosage of 468 mg/kg bw (in1% methyl cellulose) was administered by oral gavage to male and female mice. Following dosing, the animals were examined regularly for any clinical signs of reaction. Bone morrow smears were obtained at 3 sampling times: 24, 48 or 72 hours after dosing. One smear from each animal was examined for the presence of micronuclei in 1000 polychromatic erythrocytes. The ratio of polychromatic to normochromatic erythrocytes was assessed by examination of at least 1000 erythrocytes from each animal. A vehicle control (1% methylcellulose) and a psoitive control with mitomycin C by intraperitoneal injection were included. At all sampling times, mice treated with test substance showed no significant increase in the frequency of micronucleated polychromatic erythrocytes. There was no significant decrease in the ratio of polychromatic to normochromatic erythrocytes at any of the three kill times after treatment. The positive control compound, mitomycin C, produced large, highly significant increases in the frequency of micronucleated polychromatic erythrocytes together with large decreases in the ratio of polychromatic to normochromatic erythrocytes and increases in the frequency of micronucleated normochromatic erythrocytes. Under the study conditions, the test substance was found to show no evidence of clastogenic potential in the bone marrow cells of mice (Allen, 1983).