Registration Dossier

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:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Poly (diallyldimethyl ammonium chloride)
- Molecular formula (if other than submission substance): C8H16N.Cl
- Molecular weight (if other than submission substance): 161.68
- Smiles notation (if other than submission substance): C(=C)CN{+}(c)(c)(.Cl{-})CC=C
- InChl (if other than submission substance): 1/C8H16N.ClH/c1-5-7-9(3,4)8-6-2;/h5-6H,1-2,7-8H2,3-4H3;1H/q+1;/p-1
- 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 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.