reaction mass of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-2H -isothiazol-3-one

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 475 (Mammalian Bone Marrow Chromosome Aberration Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPP 84-2

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Version / remarks:

Guideline 59 Nohsan No. 4200

Deviations:

no

GLP compliance:

yes

Type of assay:

chromosome aberration assay

Test material

Test material information

Constituent 1

Test material form:

other: Amber-gold liquid

Details on test material:

- Name of test material (as cited in study report): Kathon™ 886 MW
- Physical state: Amber-gold liquid
- Stability under test conditions: Stable at room temperature

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Rohm and Haas, Batch No. J70089

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature
- Stability under test conditions: Stable at room temperature
- Solubility and stability of the test substance in the solvent/vehicle: Soluble and stable
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: No

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Dilution in water prior to oral gavage

OTHER SPECIFICS: Purity of test material was 14.1%

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 Laboratories, Portage, Michigan, USA
- Age at study initiation: 6-7 weeks
- Weight at study initiation: 21-32 g males, 20-30 g females
- Assigned to test groups randomly: Not described
- Fasting period before study: Not described
- Housing: Not described
- Diet (e.g. ad libitum): Not described
- Water (e.g. ad libitum): Not described
- Acclimation period: Not described

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: water
- Amount of vehicle (if gavage or dermal): 10 mL/kg

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: test substance in water administered by oral gavage

Duration of treatment / exposure:

n/a

Frequency of treatment:

Single oral gavage

Post exposure period:

6, 24 and 48 h after treatment

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

30 mg/kg bw (total dose)

Remarks:

Based on a.i.

Doses / concentrations 2

Dose / conc.:

15 mg/kg bw (total dose)

Remarks:

Based on a.i.

Doses / concentrations 3

Dose / conc.:

3 mg/kg bw (total dose)

Remarks:

Based on a.i.

No. of animals per sex per dose:

7/sex/group per time point for water control and high dose, low and mid doses had 5/sex/group per time point (6, 24 and 48 h time points), and positive control (TEM) had 5/sex/group and were sacrificed at 24 h only

Control animals:

yes, concurrent vehicle

Positive control(s):

Triethylenemelamine
- Route of administration: oral gavage
- Doses / concentrations: 1.0 mg/kg

Examinations

Tissues and cell types examined:

Bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: Range finding test: 10, 20, 30, 40 and 50 mg a.i./kg. High mortality at 40 and 50 mg a.i./kg.

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Animals were given a single oral dose of the test article at concentrations of 3, 15 or 30 mg a.i./kg and and euthanized at 6, 24 or 48 h after treatment

DETAILS OF SLIDE PREPARATION: Bone marrow cells from both femurs were centrifuged, fixed, spread on glass slides, air-dried, stained with Giemsa stain, and mounted with cover glasses.

METHOD OF ANALYSIS: For each animal, at least 50 metaphases were read and scored using a microscope.

Evaluation criteria:

Chromosome aberrations

Statistics:

Not described

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Remarks:

subtoxic concentrations

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: 10, 20, 30, 40 and 50 mg a.i./kg
- Clinical signs of toxicity in test animals: High mortality at 40 and 50 mg a.i./kg.

Applicant's summary and conclusion

Conclusions:

Kathon™ 886 MW biocide was negative up to 30 mg a.i./kg (didn’t cause chromosome aberrations in bone marrow cells) in this test system.

Executive summary:

OECD 475, US EPA 84-2 and Japan Guideline 59 Nohsan No. 4200. Adult CD-1 male and female mice were dosed with a single oral dose of the test article at concentrations of 3, 15 or 30 mg a.i./kg. Control animals received a single oral dose (vehicle) of distilled water or an i.p. injection of 1.0 mg/kg triethylenemelamine (TEM, positive control). Animals were euthanized at 6, 24 or 48 h after treatment. Positive control animals were euthanized only after 24 h of treatment. Bone marrow cells from both femurs were centrifuged, fixed, spread on glass slides, air-dried, stained with Giemsa stain, mounted with cover glasses. and read using a microscope.For each animal, at least 50 metaphases were scored. There were no significant increases in the number of aberrant cells in the treated groups relative to the control groups at any harvest time.