Ethyldimethyl[2-[(2-methyl-1-oxoallyl)oxy]ethyl]ammonium ethyl sulphate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: DNA damage and/or repair

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1991

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Well documented report of a guideline study conducted to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): Poly (diallyldimethyl ammonium chloride)
- Substance type: non-volatile, organic
- Physical state: aqueous solution
- Analytical purity: no data
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: no data
- Purity test date: not tested
- Lot/batch No.: EC17D9B B-08
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: room temperature, protected from light

Test animals

Species:

mouse

Strain:

ICR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Sprague Dawley, Inc, Frederick, MD
- Age at study initiation: 6-8 weeks
- Weight at study initiation:
- toxicity studies: males, 30-51 g; females, 22-30 g
- micronucleus assay: males 24-32 g; females, 20-28 g
- Assigned to test groups randomly: yes,
- Fasting period before study: no
- Housing: 5 per cage in plastic autoclavable cages with filter tops
- Diet: ad libitum (certified laboratory rodent chow)
- Water : ad libidum (tap water)
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°F): 74+/-6°F
- Humidity (%): 50 +/- 20%
- Photoperiod: 12hrs dark / 12hrs light

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

distilled water

Details on exposure:

Male and female ICR mice were exposed to Diallyldimethylammonium chloride (DADMAC), which was administered at a constant rate of 10 ml/kg as a single IP injection

Duration of treatment / exposure:

One single IP

Frequency of treatment:

once

Doses / concentrations

Remarks:

Doses / Concentrations:
22, 43 or 85 mg/kg
Basis:
analytical conc.

No. of animals per sex per dose:

15 males and 15 females per test group and vehicle. 5 males and 5 females in the positive control group

Control animals:

yes, concurrent vehicle

Positive control(s):

- Positive control: triethylenemelamine
- Route of administration: IP
- Doses / concentrations: 0.25 mg/kg

Examinations

Tissues and cell types examined:

Bone marrow and erythrocytes

Details of tissue and slide preparation:

Immediately following sacrifice, the femurs were exposed, cut just above the knee, and the bone marrow was aspirated into a syringe containing foetal bovine serum. The bone marrow cells were transferred to a capped centrifuge tube containing approximately 1 ml FBS. The bone marrow cells were pelleted by centrifugation at approximately 100 x g for five minutes and the supernatant was drawn off, leaving a small amount of serum with the remaining cell pellet. The cells were resuspended by aspiration with a capillary pipet and a small drop of bone marrow suspension was spread onto a clean glass slide. Two to four slides were prepared from each animal. The slides were fixed in methanol, stained with May-Gruenwald-Giemsa and permanently mounted. Slides were coded using a random number table by an individual not involved with the scoring process. Using medium magnification, an area of acceptable quality was selected such that the cells were well spread and stained. Using oil immersion, 1000 polychromatic erythrocytes were scored for the presence of micronuclei. The number of micronucleated normocytes in the field of 1000 polychromatic erythrocytes was enumerated. The proportion of polychromatic erythrocytes to total erythrocytes was also recorded.

Evaluation criteria:

The incidence of micronucleated polychromatic erythrocytes per 1000 polychromatic erythrocytes was determined for each animal and treatment group. The mean incidence of micronucleated polychromatic erythrocytes must not exceed 5/1000 polychromatic erythrocytes (0.5%) in the negative control (vehicle) The incidence of micronucleated polychromatic erythrocytes in the positive control group must be significantly increased relative to negative control (p<0.05, Kastenbaum-Bowman tables)

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

No significant increase in polynucleated, polychromatic ethrocytes were observed at 24, 48 or 96 hours after dose administration in males or females.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
The negative and positive controls fulfilled the requirement for determination of the valid test. Under the experimental conditions, the test substance did not increase the incidence of micronuclei in mouse bone marrow and was concluded to be negative in the micronucleus test using male and female ICR mice.